Sample records for amnion

  1. Gamma radiation sterilized amnion: use in ophthalmology

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    Martinez P, M. E. [ININ, Carretera Mexico-Toluca s/n, Ocoyoacac 52750, Estado de Mexico (Mexico); Leon T, Y. [Hospital General Regional 220, IMSS, Paseo Tollocan No. 620, Col. Vertice, Toluca 50150, Estado de Mexico (Mexico); Vazquez M, L., E-mail: esther.martinez@inin.gob.m [Hospital General de Mexico, Dr. Balmis 148, Col. Doctores, 06720 Mexico D. F. (Mexico)


    Amnion processed at the Radio sterilized Tissue Bank at the National Institute of Nuclear Research, sterilized with {sup 60}Co gamma radiation, have been used in Mexico since 2005 either as a graft to replace the damaged ocular surface, or as a patch to prevent unwanted inflammatory reactions. Patients from the Hospital General de Mexico (HGM) and Instituto Mexicano del Seguro Social (IMSS), suffering diverse pathologies such as keratoconjunctivitis; recurrent pterygium associated with symblepharon; corneal neuro trophic ulcers, chemical and thermal burns, and corneal thinning s, had been successfully treated with irradiated amnion. In the HGM, a clinical prospective study on lesions of the ocular surface of 17 eyes from 15 patients, affected with the above mentioned pathologies, was successful in 88.2%. The results have proven to be excellent as much for cosmetic purposes as for functional ones. Without the treatment, the patients could have suffered a healing after-effect or loss of sight. At IMSS, a controlled clinical randomized trial with 108 eyes from 100 patients, affected with primary nasal pterygium, was performed in 2009. These eyes were treated with radio sterilized amnion and intraoperative mitomycin C to prevent recurrence after excision of the primary pterygium. The preliminary results do not shown adverse reaction, inflammation and pain were significantly reduced radio sterilized amnion also offer security because they do no express antigens HLA-A, B or Dr and the sterile irradiated tissue do not provoke rejection or transmit an infective disease. (Author)

  2. Isolation and characterization of equine amnion mesenchymal stem cells


    Coli, Alessandra; Nocchi, Francesca; Lamanna, Roberta; Iorio, Mariacarla; Lapi, Simone; Urciuoli, Patrizia; Scatena, Fabrizio; Giannessi, Elisabetta; Stornelli, Maria Rita; Passeri, Simona


    The amnion is a particular tissue whose cells show features of multipotent stem cells proposed for use in cellular therapy and regenerative medicine. From equine amnion collected after the foal birth we have isolated MSCs (mesenchymal stem cells), namely EAMSCs (equine amnion mesenchymal stem cells), from the mesoblastic layer. The cells were grown in α-MEM (α-modified minimum essential medium) and the effect of EGF (epidermal growth factor) supplementation was evaluated. To assess the growth...

  3. Genome-Wide Analysis of DNA Methylation in Human Amnion

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    Jinsil Kim


    Full Text Available The amnion is a specialized tissue in contact with the amniotic fluid, which is in a constantly changing state. To investigate the importance of epigenetic events in this tissue in the physiology and pathophysiology of pregnancy, we performed genome-wide DNA methylation profiling of human amnion from term (with and without labor and preterm deliveries. Using the Illumina Infinium HumanMethylation27 BeadChip, we identified genes exhibiting differential methylation associated with normal labor and preterm birth. Functional analysis of the differentially methylated genes revealed biologically relevant enriched gene sets. Bisulfite sequencing analysis of the promoter region of the oxytocin receptor (OXTR gene detected two CpG dinucleotides showing significant methylation differences among the three groups of samples. Hypermethylation of the CpG island of the solute carrier family 30 member 3 (SLC30A3 gene in preterm amnion was confirmed by methylation-specific PCR. This work provides preliminary evidence that DNA methylation changes in the amnion may be at least partially involved in the physiological process of labor and the etiology of preterm birth and suggests that DNA methylation profiles, in combination with other biological data, may provide valuable insight into the mechanisms underlying normal and pathological pregnancies.

  4. The effect of gamma irradiation on injectable human amnion collagen

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    Liu, B.C.; Harrell, R.; Davis, R.H.; Dresden, M.H.; Spira, M. (Institute of Occupational Medicine, Beijing (China))


    The effect of gamma irradiation on the physicochemical properties of injectable human amnion collagen was investigated. Pepsin-extracted human amnion collagen was purified, reconstituted, and irradiated with varying doses of gamma irradiation (0.25 Mrads to 2.5 Mrads). Gamma irradiation had a significant impact on the physical characteristics of the collagen. The neutral solubility of collagen in PBS at 45{degrees}C was decreased from 100% for the nonirradiated control sample to 16% for the 2.5 Mrads irradiated sample. SDS polyacrylamide gel electrophoresis also demonstrated the dose-dependent effect of gamma irradiation on collagen cross-links. Electron microscopic observation revealed that even at low irradiation dose (0.25 Mrads), collagen fibril diameter increased. The average diameter was 50 nm for nonirradiated control fibrils, while 4.4% of the irradiated collagen fibrils had a diameter greater than 100 nm. Irradiated collagen showed little evidence of damage. Well-preserved cross-striations were found in collagen fibrils at all doses of irradiation. Native amnion collagen irradiated with gamma rays demonstrated a slight increase in resistance to collagenase degradation compared with nonirradiated native collagen samples. Increased resistance to collagenase did not correlate with increasing irradiation dose. After 30 min of incubation at 37{degrees}C, both irradiated and nonirradiated collagen was completely digested by collagenase. However, gamma-irradiated collagen did become more sensitive to hydrolysis by trypsin. The higher the irradiation doses used, the greater sensitivity to trypsin was observed. At 0.25 Mrads irradiation only a slight increase was found. No marked differences in amino acid composition were noted among the high dose irradiated, low dose irradiated and control amnion collagen.

  5. Maintenance of human amnion epithelial cell phenotype in pulmonary surfactant


    McDonald, Courtney A.; Melville, Jacqueline M; Graeme R Polglase; Jenkin, Graham; Moss, Timothy JM


    Introduction Preterm newborns often require mechanical respiratory support that can result in ventilation-induced lung injury (VILI), despite exogenous surfactant treatment. Human amnion epithelial cells (hAECs) reduce lung inflammation and resultant abnormal lung development in preterm animals; co-administration with surfactant is a potential therapeutic strategy. We aimed to determine whether hAECs remain viable and maintain function after combination with surfactant. Methods hAECs were inc...

  6. Propolis and amnion reepithelialise second-degree burns in rats. (United States)

    Pessolato, Alicia Greyce Turatti; Martins, Daniele dos Santos; Ambrósio, Carlos Eduardo; Mançanares, Celina Almeida Furlanetto; de Carvalho, Ana Flávia


    Burns are serious consequences of trauma in terms of both imminent mortality and prolonged periods of morbidity. They are often accompanied by unsatisfactory cosmetic as well as functional and psychological outcomes. These complications emphasise the need for stronger efforts in achieving greater diversity and effectiveness in the treatment of skin burns. This study aimed to verify the effectiveness of gross and microscopic epidermal and dermal responses in the process of regenerative repair or healing of burns in rats that were treated either daily with 5% propolis ointment or by autologous amnion graft. Second-degree burns were inflicted in the neck region of female rats by contact with a hot metal (at 130 °C) for 5 s. Propolis treatment accelerated the process of tissue repair and led to decreased local inflammation, which indicates that treatment with propolis was successful in the initial period (7 days) and stimulated the production of collagen fibre (assessed by morphometry) in all the periods evaluated (14 and 21 days). Amnion treatment inhibited local inflammation (assessed macroscopically), stimulated local epithelial regeneration (assessed microscopically) and stimulated the production of collagen fibre (assessed by morphometry) in the days following burn. These treatments offer new therapeutic strategies for treating severe skin burns; these strategies may allow the minimisation of scar formation, a more rapid return of function and, ultimately, a better quality of life for burn patients.

  7. A discrete network model to represent the deformation behavior of human amnion. (United States)

    Mauri, Arabella; Hopf, Raoul; Ehret, Alexander E; Picu, Catalin R; Mazza, Edoardo


    A discrete network model (DNM) to represent the mechanical behavior of the human amnion is proposed. The amnion is modeled as randomly distributed points interconnected with connector elements representing collagen crosslinks and fiber segments, respectively. This DNM is computationally efficient and allows simulations with large domains. A representative set of parameters has been selected to reproduce the uniaxial tension-stretch and kinematic responses of the amnion. Good agreement is found between the predicted and measured equibiaxial tension-stretch curves. Although the model represents the amnion phenomenologically, model parameters are physically motivated and their effect on the tension-stretch and in-plane kinematic responses is discussed. The model is used to investigate the local response in the near field of a circular hole, revealing that the kinematic response at the circular free boundaries leads to compaction and strong alignment of the network at the border of the defect. PMID:26700570

  8. Halofuginone- and Chitosan-Coated Amnion Membranes Demonstrate Improved Abdominal Adhesion Prevention

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    Scott Washburn


    Full Text Available Our objective was to determine whether coating the amniotic membrane with halofuginone, a type 1 collagen synthase inhibitor, with or without the hemostasis-inducing substance chitosan, reduced the number and severity of adhesions in the rat uterine horn injury model. Sixty retired breeder Sprague-Dawley rats underwent midline laparotomy and a zone of ischemia was created in the left uterine horn of each animal. Rats were randomized to one of six treatment groups: (1 untreated control, (2 oxidized regenerated cellulose (Interceed® (ORC, (3 plain amnion, (4 amnion coated on both sides with 0.5% solution of halofuginone (HAH, (5 amnion coated on one side with 0.5% halofuginone and on the other side with chitosan (CAH, or (6 amnion coated on both sides with chitosan (CAC. The zone of ischemia in each left uterine horn was wrapped in each treatment. Rats were sacrificed 2 weeks after laparotomy, and adhesions were counted and scored for severity. Data were analyzed using Chi square and a p <0.05 was considered significant. Our results showed that there were no differences in the percentage of animals with adhesions in the untreated, ORC, plain amnion, or CAC groups. No adhesions formed in any animal in the HAH group and only 14% of the animals developed adhesions to the uterine horn in the CAH group (p < 0.05. The percentage of animals with moderate and severe adhesions did not differ between untreated controls and the ORC groups, but were significantly reduced in all four of the amnion groups: plain amnion, HAH, CAH, and CAC (p < 0.05. Amnion coated with halofuginone alone or in combination with chitosan reduced the percentage of animals with adhesions, as well as the percentage of animals with moderate and severe adhesions compared to untreated controls and the ORC group in the rat uterine horn injury model. Amnion alone or coated with chitosan reduced the percentage of rats with moderate and severe adhesions, but not the percentage of rats with

  9. Initiation of bone and amnion banking in Turkey

    International Nuclear Information System (INIS)

    There is a growing demand in Turkey for human tissue to use in surgery and wound healing. However, our country does not have facilities for local production of tissue grafts and generally depends on imported products. Under a multi-year project initiated in 1997, the International Atomic Energy Agency (IAEA) has provided main equipment for tissue processing and experts on tissue banking as well as training on tissue processing methods. In this presentation, information on various stages of the project implementation is given. Details of lay out for the process laboratories and list of equipment supplied by the Agency and purchased locally are also given. Donor selection and testing, processing procedures for bone and amnion, setting up product design, implementation of quality system and radiation sterilisation are described briefly. Quality procedures included preparation of quality manual, Standard Operating Procedures (SOP), record forms, document control, non-conformance and corrective actions, training records, equipment maintenance and calibration are all in line with Good Manufacturing Practice (GMP) standards. Clinical applications of tissue grafts and medico-legal position of organ and tissue donation in Turkey are also discussed briefly

  10. Smad5 determines murine amnion fate through the control of bone morphogenetic protein expression and signalling levels. (United States)

    Bosman, Erika A; Lawson, Kirstie A; Debruyn, Joke; Beek, Lisette; Francis, Annick; Schoonjans, Luc; Huylebroeck, Danny; Zwijsen, An


    Smad5 is an intracellular mediator of bone morphogenetic protein (Bmp) signalling. It is essential for primordial germ cell (PGC) development, for the development of the allantois and for amnion closure, as demonstrated by loss of Bmp signalling. By contrast, the appearance of ectopic PGC-like cells and regionalized ectopic vasculogenesis and haematopoiesis in thickened Smad5(m1/m1) amnion are amnion defects that have not been associated with loss of Bmp signalling components. We show that defects in amnion and allantois can already be detected at embryonic day (E) 7.5 in Smad5 mutant mice. However, ectopic Oct4-positive (Oct4(+)) and alkaline phosphatase-positive (AP(+)) cells appear suddenly in thickened amnion at E8.5, and at a remote distance from the allantois and posterior primitive streak, suggesting a change of fate in situ. These ectopic Oct4(+), AP(+) cells appear to be Stella negative and hence cannot be called bona fide PGCs. We demonstrate a robust upregulation of Bmp2 and Bmp4 expression, as well as of Erk and Smad activity, in the Smad5 mutant amnion. The ectopic expression of several Bmp target genes in different domains and the regionalized presence of cells of several Bmp-sensitive lineages in the mutant amnion suggest that different levels of Bmp signalling may determine cell fate. Injection of rBMP4 in the exocoelom of wild-type embryos can induce thickening of amnion, mimicking the early amnion phenotype in Smad5 mutants. These results support a model in which loss of Smad5 results paradoxically in gain of Bmp function defects in the amnion. PMID:16887830

  11. Aquaporins in ovine amnion: responses to altered amniotic fluid volumes and intramembranous absorption rates. (United States)

    Cheung, Cecilia Y; Anderson, Debra F; Brace, Robert A


    Aquaporins (AQPs) are transmembrane channel proteins that facilitate rapid water movement across cell membranes. In amniotic membrane, the AQP-facilitated transfer of water across amnion cells has been proposed as a mechanism for amniotic fluid volume (AFV) regulation. To investigate whether AQPs modulate AFV by altering intramembranous absorption (IMA) rate, we tested the hypothesis that AQP gene expression in the amnion is positively correlated with IMA rate during experimental conditions when IMA rate and AFV are modified over a wide range. The relative abundances of AQP1, AQP3, AQP8, AQP9, and AQP11 mRNA and protein were determined in the amnion of 16 late-gestation ovine fetuses subjected to 2 days of control conditions, urine drainage, urine replacement, or intraamniotic fluid infusion. AQP mRNA levels were determined by RT-qPCR and proteins by western immunoblot. Under control conditions, mRNA levels among the five AQPs differed more than 20-fold. During experimental treatments, mean IMA rate in the experimental groups ranged from 100 ± 120 mL/day to 1370 ± 270 mL/day. The mRNA levels of the five AQPs did not change from control and were not correlated with IMA rates. The protein levels of AQP1 were positively correlated with IMA rates (r(2) = 38%, P = 0.01) while the remaining four AQPs were not. These findings demonstrate that five AQPs are differentially expressed in ovine amnion. Our study supports the hypothesis that AQP1 may play a positive role in regulating the rate of fluid transfer across the amnion, thereby participating in the dynamic regulation of AFV.

  12. The Use of Amnion-Derived Cellular Cytokine Solution to Improve Healing in Acute and Chronic Wound Models


    Franz, Michael G.; Payne, Wyatt G.; Xing, Liyu; Naidu, D. K; Salas, R. E; Marshall, Vivienne S.; Trumpower, C. J; Smith, Charlotte A; Steed, David L.; Robson, M. C.


    Objective: Growth factors demonstrate mixed results improving wound healing. Amnion-derived multipotent cells release physiologic levels of growth factors and tissue inhibitors of metalloproteinases. This solution was tested in models of acute and chronic wound healing. Methods: Acute model: Sprague-Dawley rats underwent laparotomy incisions. The midline fascia was primed with phosphate-buffered saline, unconditioned media, or amnion-derived cellular cytokine suspension prior to incision. Bre...

  13. Safety Profile of Amnion-Derived Cellular Cytokine Solution (ACCS) Following Topical Skin Application in Patients Receiving Breast Radiotherapy


    Trombetta, Mark; Julian, Thomas B.; Wickerham, D. Lawrence; Steed, David L.


    Objective: To establish a safety profile for amnion-derived cellular cytokine solution following topical application in patients undergoing whole breast radiotherapy for breast cancer. Materials and Methods: Twenty female patients with early-stage breast cancer were enrolled in 2 separate cohorts of an institutional review board–approved phase I protocol. Cohort 1 consisted of 10 patients who received topical amnion-derived cellular cytokine solution to the breast immediately following the fi...

  14. Amnion and Chorion Membranes: Potential Stem Cell Reservoir with Wide Applications in Periodontics


    Akanksha Gupta; Suresh D. Kedige; Kanu Jain


    The periodontal therapy usually aims at elimination of disease causing bacteria and resolution of inflammation. It involves either resective or regenerative surgery to resolve the inflammation associated defects. Over the years, several methods have been used for achievement of periodontal regeneration. One of the oldest biomaterials used for scaffolds is the fetal membrane. The amniotic membranes of developing embryo, that is, amnion (innermost lining) and chorion (a layer next to it), have ...

  15. Nuclear factor kappa B activation occurs in the amnion prior to labour onset and modulates the expression of numerous labour associated genes.

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    Sheri Lim

    Full Text Available BACKGROUND: Prior to the onset of human labour there is an increase in the synthesis of prostaglandins, cytokines and chemokines in the fetal membranes, particular the amnion. This is associated with activation of the transcription factor nuclear factor kappa B (NFκB. In this study we characterised the level of NFκB activity in amnion epithelial cells as a measure of amnion activation in samples collected from women undergoing caesarean section at 39 weeks gestation prior to the onset of labour. METHODOLOGY/PRINCIPAL FINDINGS: We found that a proportion of women exhibit low or moderate NFκB activity while other women exhibit high levels of NFκB activity (n = 12. This activation process does not appear to involve classical pathways of NFκB activation but rather is correlated with an increase in nuclear p65-Rel-B dimers. To identify the full range of genes upregulated in association with amnion activation, microarray analysis was performed on carefully characterised non-activated amnion (n = 3 samples and compared to activated samples (n = 3. A total of 919 genes were upregulated in response to amnion activation including numerous inflammatory genes such cyclooxygenase-2 (COX-2, 44-fold, interleukin 8 (IL-8, 6-fold, IL-1 receptor accessory protein (IL-1RAP, 4.5-fold, thrombospondin 1 (TSP-1, 3-fold and, unexpectedly, oxytocin receptor (OTR, 24-fold. Ingenuity Pathway Analysis of the microarray data reveal the two main gene networks activated concurrently with amnion activation are i cell death, cancer and morphology and ii cell cycle, embryonic development and tissue development. CONCLUSIONS/SIGNIFICANCE: Our results indicate that assessment of amnion NFκB activation is critical for accurate sample classification and subsequent interpretation of data. Collectively, our data suggest amnion activation is largely an inflammatory event that occurs in the amnion epithelial layer as a prelude to the onset of labour.

  16. Prostaglandin E2 regulation of amnion cell vascular endothelial growth factor expression: relationship with intramembranous absorption rate in fetal sheep. (United States)

    Cheung, Cecilia Y; Beardall, Michael K; Anderson, Debra F; Brace, Robert A


    We hypothesized that prostaglandin E2 (PGE2) stimulates amniotic fluid transport across the amnion by upregulating vascular endothelial growth factor (VEGF) expression in amnion cells and that amniotic PGE2 concentration correlates positively with intramembranous (IM) absorption rate in fetal sheep. The effects of PGE2 at a range of concentrations on VEGF 164 and caveolin-1 gene expressions were analyzed in cultured ovine amnion cells. IM absorption rate, amniotic fluid (AF) volume, and PGE2 concentration in AF were determined in late-gestation fetal sheep during control conditions, isovolumic fetal urine replacement (low IM absorption rate), or intra-amniotic fluid infusion (high IM absorption rate). In ovine amnion cells, PGE2 induced dose- and time-dependent increases in VEGF 164 mRNA levels and reduced caveolin-1 mRNA and protein levels. VEGF receptor blockade abolished the caveolin-1 response, while minimally affecting the VEGF response to PGE2. In sheep fetuses, urine replacement reduced amniotic PGE2 concentration by 58%, decreased IM absorption rate by half, and doubled AF volume (P amniotic fluid infusion increased IM absorption rate and AF volume (P amniotic PGE2 concentration was unchanged. Neither IM absorption rate nor AF volume correlated with amniotic PGE2 concentration under each experimental condition. Although PGE2 at micromolar concentrations induced dose-dependent responses in VEGF and caveolin-1 gene expression in cultured amnion cells consistent with a role of PGE2 in activating VEGF to mediate AF transport across the amnion, amniotic PGE2 at physiological nanomolar concentrations does not appear to regulate IM absorption rate or AF volume.

  17. Use of gamma-irradiated amnion as a biological dressing in the management of radiation induced ulcers

    International Nuclear Information System (INIS)

    Full text: The treatment of moist skin desquamation (ulceration) following radiation therapy focuses on promoting healing, preventing microbial infections, protecting from mechanical trauma and providing pain relief. Various dressings like hydrocolloid, 1% gentian violet and other topical applications are routinely used. This study evaluated the use of freeze-dried, irradiated amnion from the Tata Memorial Hospital Tissue Bank in the management of radiation ulcers from 2001 to 2007. During this period, 2095 amnion dressings were used in 554 patients (238 male and 316 female). The ulcer was cleaned with normal saline and the amnion applied directly on the affected area. No topical antibiotics were used. The ulcer sites were in the regions of the groin and perianal area (40.79%), head and neck (30.50%), breast and axilla (18.59%), limbs (5.78%), chest (2.70%) and back (1.62%). Healing occurred within 4 days to 14 days using a single application of amnion in 291 patients (52.53%), 2 dressings in 129 patients (23.28%) and 3 dressings in 74 patients (13.36%). The remaining 60 patients (10.83%) required four or more dressings which were used over a period of a month. The amnion dressing was found to be convenient to apply and adhered well to the affected areas without the use of any adhesives unlike with routine hydro-colloid dressing. This was particularly advantageous in sites with irregular contours such as the groin and perianal areas and the head and neck regions (71.29%). Patients experienced almost immediate pain relief and the need for analgesics was reduced. In the majority of patients healing occurred earlier than was usually obtained with routine dressings. The amnion dressing proved to be advantageous over co ventional dressings because of its convenience of use and cost effectiveness due to shorter duration of healing, fewer dressing changes and diminished use of analgesics and antibiotics. (Author)

  18. Nuclear microanalysis of the human amnion: A study of ionic cellular exchanges (United States)

    Razafindrabe, L.; Moretto, Ph.; Llabador, Y.; Simonoff, M.; Bara, M.; Guiet-Bara, A.


    The epithelial cells of the human amniotic membrane have been extensively studied by electrophysiologists with the aim of elucidating the mechanisms of transmembrane ionic transfers. In order to provide complementary information about this model, nuclear microanalysis was performed using the CENBG ion microbeam. Quantitative mapping of the human amnion was carried out and the distributions of most mono- and divalent ions involved in cellular pathways (Na +, Mg 2+, Cl -, Ca 2+) were determined. The ionic cellular content was also compared, before and after incubation in a Hanks' physiological fluid and the resultant ions transfers were determined. The aim of this paper is to expose the advances of this experimental model, more particularly after the development of simulation programs which improved the accuracy of PIXE analysis in the measurement of low energy X-rays emitters. Statistically significant results can now be extracted and can be explained taking into account the results of previous electrophysiological experiments.

  19. Isolation of Human Amnion Epithelial Cells According to Current Good Manufacturing Procedures. (United States)

    Gramignoli, Roberto; Srinivasan, Raghuraman C; Kannisto, Kristina; Strom, Stephen C


    Different cell types can be isolated from human placental tissues, and some have been reported to retain phenotypic plasticity and characteristics that make them a promising source of cells for regenerative medicine. Among these are human amnion epithelial cells (hAECs). Adoption of current good manufacturing practices (cGMP) and enhanced quality control is essential when isolating hAECs in order to deliver a safe and effective cellular product for clinical purposes. This unit describes a detailed protocol for selective isolation of hAECs from human term placenta with little to no contamination by other cell types. A method for characterizing the heterogeneity of the hAEC suspension is also provided. The resulting cell product will be useful for clinical as well as basic research applications. © 2016 by John Wiley & Sons, Inc. PMID:27171794

  20. Human amnion epithelial cells can be induced to differentiate into functional insulin-producing cells

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    Yanan Hou; Qin Huang; Tianjin Liu; Lihe Guo


    Pancreatic islet transplantation has demonstrated that long-term insulin independence may be achieved in patients suffering from diabetes mellitus type 1. However, limited availability of islet tissue means that new sources of insulinproducing cells that are responsive to glucose are required. Here, we show that human amnion epithelial cells (HAEC) can be induced to differentiate into functional insulinproducing cells in vitro. After induction of differentiation, HAEC expressed multiple pancreatic --cell genes, including insulin, pancreas duodenum homeobox-1, paired box gene 6,NK2 transcription factor-related locus 2, Islet 1, glucokinase,and glucose transporter-2, and released C-peptide in a glucose-regulated manner in response to other extracellular stimulations. The transplantation of induced HAEC into streptozotocin-induced diabetic C57 mice reversed hyperglycemia, restored body weight, and maintained euglycemia for 30 d. These findings indicated that HAEC may be a new source for cell replacement therapy in type 1 diabetes.

  1. The dynamic expression of extraembryonic marker genes in the beetle Tribolium castaneum reveals the complexity of serosa and amnion formation in a short germ insect. (United States)

    Sharma, Rahul; Beermann, Anke; Schröder, Reinhard


    Most insect embryos develop with two distinct extraembryonic membranes, the serosa and the amnion. In the insect beetle Tribolium the early origin of the serosa within the anterior blastoderm is well established but the origin of the amnion is still debated. It is not known whether this tissue develops from a blastodermal precursor or originates de novo later from embryonic tissue during embryogenesis. We undertook an in-depth analysis of the spatio-temporal expression pattern profile of important extraembryonic membrane marker genes with emphasis on early blastoderm development in Tribolium. The amnion marker iroquois (Tc-iro) was found co-expressed with the serosa marker zerknüllt1 (Tc-zen1) during early blastoderm formation in an anterior cap domain. This domain later resolved into two adjacent domains that likely represent the precursors of the serosa and the amnion. In addition, we found the hindsight ortholog in Tribolium (Tc-hnt) to be a serosa-specific marker. Surprisingly, decapentaplegic (Tc-dpp) expression was not seen as a symmetric cap domain but detected asymmetrically first along the DV- and later also along the AP-axis. Moreover, we found a previously undescribed domain of phosphorylated MAD (pMAD) protein in anterior ventral serosal cells. This is the first study showing that the anterior-lateral part of the amnion originates from the anterior blastoderm while the precursor of the dorsal amnion develops later de novo from a dorsal-posterior region within the differentiated blastoderm.

  2. Mesenchymal stem cells from amnion and amniotic fluid in the bovine. (United States)

    Corradetti, B; Meucci, A; Bizzaro, D; Cremonesi, F; Lange Consiglio, A


    Amnion and amniotic fluid (AF) are noncontroversial and inexhaustible sources of mesenchymal stem cells (MSCs) that can be harvested noninvasively at low cost. As in humans, also in veterinary field, presumptive stem cells derived from these tissues reveal as promising candidates for disease treatment, specifically for their plasticity, their reduced immunogenicity, and high anti-inflammatory potential. The aim of this work is to obtain and characterize, for the first time in bovine species, presumptive MSCs from the epithelial portion of the amnion (AECs) and from the AF (AF-MSCs) to be used for clinical applications. AECs display a polygonal morphology, whereas AF-MSCs exhibit a fibroblastic-like morphology only starting from the second passage, being heterogeneous during the primary culture. For both lines, the proliferative ability has been found constant over the ten passages studied and AECs show a statistically lower (P<0.05) doubling time with respect to AF-MSCs. AECs express MSC-specific markers (ITGB1 (CD29), CD44, ALCAM (CD166), ENG (CD105), and NT5E (CD73)) from P1 to P3; in AF-MSCs, only ITGB1, CD44, and ALCAM mRNAs are detected; NT5E is expressed from P2 and ENG has not been found at any passage. AF-MSCs and AECs are positive for the pluripotent markers (POU5F1 (OCT4) and MYC (c-Myc)) and lack of the hematopoietic markers. When appropriately induced, both cell lines are capable of differentiating into ectodermal and mesodermal lineages. This study contributes to reinforce the emerging importance of these cells as ideal tools in veterinary medicine. A deeper evaluation of the immunological properties needs to be performed in order to better understand their role in cellular therapy.

  3. Versuche zur elektrophysiologischen Charakterisierung des Amnions von Hühnerembryonen ( Gallus gallus f. domestica ) mit Hilfe der Ussing - Kammer - Methode


    Blasius, Heiner


    It was the aim of this study to characterize the chick amnion electrophysiologically using “Ussing” - chamber experiments. To this end, it was first necessary to establish an appropriate time window for the examinations. According to literature data and preliminary experimental data, incubation days D9 and D10 were chosen. Furthermore, an adequate preparation technique was established. The experimental buffer system was assembled according to literature data. After clarifying these met...

  4. Angiogenic properties of dehydrated human amnion/chorion allografts: therapeutic potential for soft tissue repair and regeneration


    Koob, Thomas J; Lim, Jeremy J.; Massee, Michelle; Zabek, Nicole; Rennert, Robert; Gurtner, Geoffrey; Li, William W


    Background Chronic wounds are associated with a number of deficiencies in critical wound healing processes, including growth factor signaling and neovascularization. Human-derived placental tissues are rich in regenerative cytokines and have been shown in randomized clinical trials to be effective for healing chronic wounds. In this study, PURION® Processed (MiMedx Group, Marietta, GA) dehydrated human amnion/chorion membrane tissue allografts (dHACM, EpiFix®, MiMedx) were evaluated for prope...

  5. 单层辐射人羊膜对皮损创面的促愈作用%Effect of single-layer radiation-treated human amnion on healing of skin wound surface

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    杨国华; 苗文哲; 刘彤; 赵薇; 孟庆刚; 贾继峰


    @@ INTRODUCTION Fresh human amnion used to cover burn wound surface has been reported, but its preparation and store is complicated that make it difficult for clinical application. Clinical application of single-layer radiation-treated human amnion in skin-offering and abrasion wound has achieved a good effect.

  6. Human amnion epithelial cells induced to express functional cystic fibrosis transmembrane conductance regulator.

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    Sean V Murphy

    Full Text Available Cystic fibrosis, an autosomal recessive disorder caused by a mutation in a gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR, remains a leading cause of childhood respiratory morbidity and mortality. The respiratory consequences of cystic fibrosis include the generation of thick, tenacious mucus that impairs lung clearance, predisposing the individual to repeated and persistent infections, progressive lung damage and shortened lifespan. Currently there is no cure for cystic fibrosis. With this in mind, we investigated the ability of human amnion epithelial cells (hAECs to express functional CFTR. We found that hAECs formed 3-dimensional structures and expressed the CFTR gene and protein after culture in Small Airway Growth Medium (SAGM. We also observed a polarized CFTR distribution on the membrane of hAECs cultured in SAGM, similar to that observed in polarized airway cells in vivo. Further, hAECs induced to express CFTR possessed functional iodide/chloride (I(-/Cl(- ion channels that were inhibited by the CFTR-inhibitor CFTR-172, indicating the presence of functional CFTR ion channels. These data suggest that hAECs may be a promising source for the development of a cellular therapy for cystic fibrosis.

  7. Characterization of Side Cell Populations Obtained from Human Amnion Mesenchymal Cells

    Institute of Scientific and Technical Information of China (English)

    LI Ning; PIAO Zhengfu; Mamoru Kobayashi; Koji Sasaki; DING Shu-qin; Aiko Kikuchi; Isao Kamo; Norio Sakuragawa


    Human amnion mesenchymal cells (AMCs) contain multipotent cells. To enrich such multipotent stem cells, we applied to AMCs the new method for the isolation of side population (SP) cells used for the enrichment of multipotent stem cells from many tissues. We succeeded in obtaining SP cells from AMCs (AMC-SP cells). AMC-SP cells were found in 0.2% of AMCs, irrespective of the length of pregnant period, ranging from 37 to 40 weeks. Cell cycle analyses uggested that AMC-SP cells belonged to a cell population that proliferated very slowly and/or was in a quiescent state in the amniotic membrane. Upon culturing, they proliferated with 40 to 80 cell doublings. However, they did not form colonies in a soft agarose culture, whereas HepG2 cells, representative human hepatoma cells formed many large colonies. These results suggest that AMC-SP cells that have considerable value for the use of regenerative medicine can be managed safely in vitro.

  8. Derivation and characterization of human embryonic stem cells on human amnion epithelial cells. (United States)

    Lai, Dongmei; Wang, Yongwei; Sun, Jian; Chen, Yifei; Li, Ting; Wu, Yi; Guo, Lihe; Wei, Chunsheng


    Culture conditions that support the growth of undifferentiated human embryonic stem cells (hESCs) have already been established using primary human amnion epithelial cells (hAECs) as an alternative to traditional mitotically inactivated mouse embryonic fibroblasts (MEFs). In the present work, inner cell masses (ICM) were isolated from frozen embryos obtained as donations from couples undergoing in vitro fertilization (IVF) treatment and four new hESC lines were derived using hAECs as feeder cells. This feeder system was able to support continuous growth of what were, according to their domed shape and markers, undifferentiated naïve-like hESCs. Their pluripotent potential were also demonstrated by embryoid bodies developing to the expected three germ layers in vitro and the productions of teratoma in vivo. The cell lines retained their karyotypic integrity for over 35 passages. Transmission electron microscopy (TEM) indicated that these newly derived hESCs consisted mostly of undifferentiated cells with large nuclei and scanty cytoplasm. The new hESCs cultured on hAECs showed distinct undifferentiated characteristics in comparison to hESCs of the same passage maintained on MEFs. This type of optimized culture system may provide a useful platform for establishing clinical-grade hESCs and assessing the undifferentiated potential of hESCs.

  9. Cases of limb-body wall complex: Early amnion rupture, vascular disruption, or abnormal splitting of the embryo? (United States)

    Crespo, Frank; Pinar, Halit; Kostadinov, Stefan


    We report two cases of limb-body wall complex (LBWC), also known as body stalk anomaly, a rare form of body wall defect incompatible with life. The first case was identified during a level II ultrasound examination performed at 7 wk gestational age. The delivery was by breech extraction at 39 wk and 4 days. The second case was delivered by spontaneous vaginal delivery at 35 wk and 5 days. Karyotype analysis was normal in both fetuses. The phenotype of LBWC is variable, but commonly identified features include: exencephaly, limb defects, and either facial clefts or thoraco-abdominoschisis. The exact etiology remains uncertain, as the disorder has been regarded as sporadic with low recurrence. Vascular disruption during early embryogenesis, early amnion rupture, abnormal splitting of the embryo, and failure of amnion fusion have been implicated in the pathogenesis of LBWC. A role for possible gene mutation and maternal use of alcohol, tobacco, or illicit drugs has also been suggested. Detailed ultrasonography along with biochemical screening may allow for early detection.

  10. Amniotic membrane is a potential regenerative option for chronic non-healing wounds: a report of five cases receiving dehydrated human amnion/chorion membrane allograft. (United States)

    Mrugala, Andrew; Sui, Audrey; Plummer, Malgorzata; Altman, Igor; Papineau, Elaine; Frandsen, Devn; Hill, Danielle; Ennis, William J


    A case series of five patients with a total of six chronic non-healing wounds (>30 day duration) were non-randomly selected to evaluate the performance, safety and handling properties of dehydrated human amnion/chorion membrane allograft, an amniotic membrane scaffolding product. The patients had lower extremity wounds that had previously failed standard of care within a university outpatient/inpatient wound healing programme. Five wounds treated with dehydrated amnion/chorion membrane allograft showed a mean 43% area reduction from baseline (51% median) at 3 weeks into treatment and completely healed with a 64-day median time to closure (SD ±27·6 days). One wound worsened at 3 weeks and was found to have a complete central vein obstruction that was treated with long-term mild compression but still eventually healed at 6 months. Removing this outlier, the four responding wounds had a 72% mean and 69% median change in area from baseline, at the 3 week point. All five patients received only one application of dehydrated human amnion/chorion membrane allograft, and there were no adverse events. The product was easy to use, administer and handle. In summary, dehydrated human amnion/chorion membrane allograft appears to be a safe, effective and easy to use therapy for chronic non-healing wounds. This study describes the details of these clinical cases and provides an overview of the current evidence on the use of amniotic tissue in clinical practice. PMID:25974156

  11. Human amnion membrane matrix as a substratum for axonal regeneration in the central nervous system. (United States)

    Gage, F H; Blaker, S N; Davis, G E; Engvall, E; Varon, S; Manthorpe, M


    Human Amnion Membrane Matrix (HAMM) was used as a substratum for the regeneration of neuronal axons in the central nervous system. A large piece of HAMM was bound to nitrocellulose paper (NCP) as a supporting material, and cut into small strips. Aspirative lesions of the fimbria-fornix were made in adult rats leaving a cavity separating the septum from the hippocampus. In the same operation a small piece of HAMM-NCP was placed into the cavity with one end abutting the axotomized septum and the other end abutting the denervated hippocampus. At times between 2 weeks and 3 months after surgery the brains of the animals were examined histologically for 1) host response to the implant; 2) maintenance of HAMM-NCP in the originally implanted orientation; 3) growth of acetylcholinesterase (AChE)-positive fibers on the HAMM; 4) growth of the AChE-positive fibers into the denervated hippocampus; and 5) non-neuronal cells on the HAMM. The NCP remained in place over the 3-month period. In most cases the HAMM and NCP remained apposed and caused no greater reaction in the brain beyond that created in response to the aspirative lesion alone. AChE-positive fibers grew out from the septum onto the HAMM by 2 weeks, and by 8 weeks more extensive growth was observed on the HAMM. By 8-weeks, AChE fibers could be clearly seen coming off the membrane and entering the host hippocampus. By double-labelling for the basement membrane side of the HAMM (using anti-human laminin antibodies) and for cholinergic fibers (using AChE histochemistry) in the same sections, it was possible to see fibers clearly growing on the laminin-positive side of the HAMM but little or no growth on the opposing laminin-negative stromal side. The most important variables for extensive growth appeared to be the accurate placement of the implant and the amount of time following the lesions. The placement of the matrix on NCP prior to implantation in the brain made it easier to orient the membrane between the septum

  12. Amnion-Epithelial-Cell-Derived Exosomes Demonstrate Physiologic State of Cell under Oxidative Stress. (United States)

    Sheller, Samantha; Papaconstantinou, John; Urrabaz-Garza, Rheanna; Richardson, Lauren; Saade, George; Salomon, Carlos; Menon, Ramkumar


    At term, the signals of fetal maturity and feto-placental tissue aging prompt uterine readiness for delivery by transitioning quiescent myometrium to an active stage. It is still unclear how the signals reach the distant myometrium. Exosomes are a specific type of extracellular vesicle (EVs) that transport molecular signals between cells, and are released from a wide range of cells, including the maternal and fetal cells. In this study, we hypothesize that i) exosomes act as carriers of signals in utero-placental compartments and ii) exosomes reflect the physiologic status of the origin cells. The primary aims of this study were to determine exosomal contents in exosomes derived from primary amnion epithelial cells (AEC). We also determined the effect of oxidative stress on AEC derived exosomal cargo contents. AEC were isolated from amniotic membrane obtained from normal, term, not in labor placentae at delivery, and culture under standard conditions. Oxidative stress was induced using cigarette smoke extract for 48 hours. AEC-conditioned media were collected and exosomes isolated by differential centrifugations. Both growth conditions (normal and oxidative stress induced) produced cup shaped exosomes of around 50 nm, expressed exosomes enriched markers, such as CD9, CD63, CD81 and HSC70, embryonic stem cell marker Nanog, and contained similar amounts of cell free AEC DNA. Using confocal microscopy, the colocalization of histone (H) 3, heat shock protein (HSP) 70 and activated form of pro-senescence and term parturition associated marker p38 mitogen activated protein kinase (MAPK) (P-p38 MAPK) co-localized with exosome enrich marker CD9. HSP70 and P-p38 MAPK were significantly higher in exosomes from AEC grown under oxidative stress conditions than standard conditions (pmass spectrometry and bioinformatics analysis identified 221 different proteins involved in immunomodulatory response and cell-to-cell communication. This study determined AEC exosome

  13. Amnion-Epithelial-Cell-Derived Exosomes Demonstrate Physiologic State of Cell under Oxidative Stress (United States)

    Sheller, Samantha; Papaconstantinou, John; Urrabaz-Garza, Rheanna; Richardson, Lauren; Saade, George; Salomon, Carlos; Menon, Ramkumar


    At term, the signals of fetal maturity and feto-placental tissue aging prompt uterine readiness for delivery by transitioning quiescent myometrium to an active stage. It is still unclear how the signals reach the distant myometrium. Exosomes are a specific type of extracellular vesicle (EVs) that transport molecular signals between cells, and are released from a wide range of cells, including the maternal and fetal cells. In this study, we hypothesize that i) exosomes act as carriers of signals in utero-placental compartments and ii) exosomes reflect the physiologic status of the origin cells. The primary aims of this study were to determine exosomal contents in exosomes derived from primary amnion epithelial cells (AEC). We also determined the effect of oxidative stress on AEC derived exosomal cargo contents. AEC were isolated from amniotic membrane obtained from normal, term, not in labor placentae at delivery, and culture under standard conditions. Oxidative stress was induced using cigarette smoke extract for 48 hours. AEC-conditioned media were collected and exosomes isolated by differential centrifugations. Both growth conditions (normal and oxidative stress induced) produced cup shaped exosomes of around 50 nm, expressed exosomes enriched markers, such as CD9, CD63, CD81 and HSC70, embryonic stem cell marker Nanog, and contained similar amounts of cell free AEC DNA. Using confocal microscopy, the colocalization of histone (H) 3, heat shock protein (HSP) 70 and activated form of pro-senescence and term parturition associated marker p38 mitogen activated protein kinase (MAPK) (P-p38 MAPK) co-localized with exosome enrich marker CD9. HSP70 and P-p38 MAPK were significantly higher in exosomes from AEC grown under oxidative stress conditions than standard conditions (pexosome characteristics and their cargo reflected the physiologic status of the cell of origin and suggests that AEC-derived exosomal p38 MAPK plays a major role in determining the fate of pregnancy

  14. Modelling the influence of amnionicity on the severity of twin-twin transfusion syndrome in monochorionic twin pregnancies

    Energy Technology Data Exchange (ETDEWEB)

    Wijngaard, Jeroen P H M van den [Laser Center and Department of Obstetrics and Gynecology, Academic Medical Center, University of Amsterdam, Amsterdam (Netherlands); Umur, Asli [Laser Center and Department of Obstetrics and Gynecology, Academic Medical Center, University of Amsterdam, Amsterdam (Netherlands); Ross, Michael G [Department of Obstetrics and Gynecology, Harbor University of California-Los Angeles Medical Center, Torrance, CA 90502 (United States); Gemert, Martin J C van [Laser Center and Department of Obstetrics and Gynecology, Academic Medical Center, University of Amsterdam, Amsterdam (Netherlands)


    Clinical treatment for diamniotic-monochorionic twin-twin transfusion syndrome (TTTS) may include conversion of diamniotic pregnancies to a monoamniotic-monochorionic state by disrupting the amnion septum. We sought to test the underlying hypothesis, i.e. that a monoamniotic state reduces the severity of TTTS. With use of our previously developed mathematical model of two equal fetoplacental circulatory units connected by various sizes and types of placental anastomoses, we compared the haemodynamic and amniotic fluid dynamics of monoamniotic and diamniotic twins that develop TTTS. We used three anastomotic patterns that produce severe, moderate or mild forms of TTTS, respectively, in our diamniotic-monochorionic twin model. Monoamnionicity was modelled by adding the two amniotic fluid volumes and using the volume-averaged amniotic fluid osmolality. The results were as follows: for severe TTTS, small differences develop between diamniotic and monoamniotic donor twins in fetal urine production, swallowed volume, blood volume, blood pressures, net fetofetal transfusion, and blood and amniotic fluid osmolality. However, the circulatory imbalance between the monoamniotic twins deteriorates similar to that of diamniotic twins. The pathophysiological differences tend to disappear for milder TTTS. In conclusion, our model suggests that the uncommon finding of TTTS in monoamniotic twins is not due to the presence of a single amniotic sac. Rather, clinically significant differences in anastomotic patterns and the delayed or lack of identification of manifestations in monoamniotic twins account for the reduced rate of TTTS diagnosis. Based on these results we expect the clinical disruption of the amnion septum in diamniotic-monochorionic TTTS pregnancies to have only minimal benefits. (note)

  15. Human amnion mesenchymal stem cells promote proliferation and osteogenic differentiation in human bone marrow mesenchymal stem cells. (United States)

    Wang, Yuli; Yin, Ying; Jiang, Fei; Chen, Ning


    Human amnion mesenchymal stem cells (HAMSCs) can be obtained from human amniotic membrane, a highly abundant and readily available tissue. HAMSC sources present fewer ethical issues, have low immunogenicity, anti-inflammatory properties, considerable advantageous characteristics, and are considered an attractive potential treatment material in the field of regenerative medicine. We used a co-culture system to determine whether HAMSCs could promote osteogenesis in human bone marrow mesenchymal stem cells (HBMSCs). We isolated HAMSCs from discarded amnion samples and collected them using pancreatin/collagenase digestion. We cultured HAMSCs and HBMSCSs in basal medium. Activity of alkaline phosphatase (ALP), an early osteogenesis marker, was increased in the co-culture system compared to the control single cultures, which we also confirmed by ALP staining. We used immunofluorescence testing to investigate the effects of co-culturing with HAMSCs on HBMSC proliferation, which revealed that the co-culturing enhanced EdU expression in HBMSCs. Western blotting and quantitative real-time PCR indicated that co-culturing promoted osteogenesis in HBMSCs. Furthermore, Alizarin red S staining revealed that extracellular matrix calcium levels in mineralized nodule formation produced by the co-cultures were higher than that in the controls. Using the same co-culture system, we further observed the effects of HAMSCs on osteogenic differentiation in primary osteoblasts by Western blotting, which better addressed the mechanism for HAMSCs in bone regeneration. The results showed HAMSCs are osteogenic and not only play a role in promoting HBMSC proliferation and osteogenic differentiation but also in osteoblasts, laying the foundation for new regenerative medicine methods.

  16. Timing of Histologic Progression from Chorio-Deciduitis to Chorio-Deciduo-Amnionitis in the Setting of Preterm Labor and Preterm Premature Rupture of Membranes with Sterile Amniotic Fluid.

    Directory of Open Access Journals (Sweden)

    Chan-Wook Park

    Full Text Available Histologic chorio-deciduitis and chorio-deciduo-amnionitis (amnionitis in extra-placental membranes are known to represent the early and advanced stages of ascending intra-uterine infection. However, there are no data in humans about the time required for chorio-deciduitis to develop and for chorio-deciduitis without amnionitis to progress to chorio-deciduitis with amnionitis, and the effect of prolongation of pregnancy on the development of chorio-deciduitis and amnionitis in patients with preterm labor and intact membranes (PTL and preterm premature rupture of membranes (preterm-PROM. We examined these issues in this study.The study population consisted of 289 women who delivered preterm (133 cases with PTL, and 156 cases with preterm-PROM and who had sterile amniotic fluid (AF defined as a negative AF culture and the absence of inflammation as evidenced by a matrix metalloproteinase-8 (MMP-8 level <23 ng/ml. We examined the association between amniocentesis-to-delivery interval and inflammatory status in the extra-placental membranes (i.e., inflammation-free extra-placental membranes, choroi-deciduitis only, and chorio-deciduitis with amnionitis in patients with PTL and preterm-PROM.Amniocentesis-to-delivery interval was longer in cases of chorio-deciduitis with amnionitis than in cases of chorio-deciduitis only in both PTL (median [interquartile-range (IQR]; 645.4 [319.5] vs. 113.9 [526.9] hours; P = 0.005 and preterm-PROM (131.3 [135.4] vs. 95.2 [140.5] hours; P<0.05. Amniocentesis-to-delivery interval was an independent predictor of the development of both chorio-deciduitis and amnionitis after correction for confounding variables such as gestational age at delivery in the setting of PTL, but not preterm-PROM.These data confirm for the first time that, in cases of both PTL and preterm-PROM with sterile AF, more time is required to develop chorio-deciduitis with amnionitis than chorio-deciduitis alone in extra-placental membranes. Moreover

  17. A roentgenographic assessment of regenerative efficacy of bioactive Gengigel® in conjunction with amnion membrane in grade II furcation defect

    Directory of Open Access Journals (Sweden)

    S Harveen Kalra


    Full Text Available Background: Nowadays, techniques are being developed to guide and instruct the specialized cellular components of the periodontium to participate in the regenerative process. This approach of reconstruction makes use of understanding of the development of the periodontium and the cellular processes that are involved. Hyaluronic acid is a naturally occurring non-sulfated high molecular weight glycosaminoglycan that forms a critical component of the extracellular matrix and contributes significantly to tissue hydrodynamics, cell migration, and proliferation. Hence, its administration to periodontal wound sites could achieve comparable beneficial effects in periodontal tissue regeneration. Hence, the purpose of the present case report was to assess roentgenographically, the regenerative capacity of Gengigel® in conjunction with bioactive amnion guided tissue regeneration (GTR membrane in a patient with Grade II furcation defect. Case Presentation: A patient complained of bleeding gums from the lower back tooth region, reportedly found Grade II furcation in the lower right mandibular first molar. After Phase, I therapy, Gengigel® along with bioactive amnion membrane was placed in the furcation area during the surgical phase. Roentgenographic assessment was done at 4 months and 6 months postoperatively. It resulted in complete defect-fill and loss of radiolucency at 6 months. Conclusion: Surgical placement of Gengigel® along with amnion membrane in the furcation defect can significantly improve the periodontal defect morphology.

  18. Comparison of structural changes in skin and amnion tissue grafts for transplantation induced by gamma and electron beam irradiation for sterilization. (United States)

    Mrázová, H; Koller, J; Kubišová, K; Fujeríková, G; Klincová, E; Babál, P


    Sterilization is an important step in the preparation of biological material for transplantation. The aim of the study is to compare morphological changes in three types of biological tissues induced by different doses of gamma and electron beam radiation. Frozen biological tissues (porcine skin xenografts, human skin allografts and human amnion) were irradiated with different doses of gamma rays (12.5, 25, 35, 50 kGy) and electron beam (15, 25, 50 kGy). Not irradiated specimens served as controls. The tissue samples were then thawn and fixed in 10 % formalin, processed by routine paraffin technique and stained with hematoxylin and eosin, alcian blue at pH 2.5, orcein, periodic acid Schiff reaction, phosphotungstic acid hematoxylin, Sirius red and silver impregnation. The staining with hematoxylin and eosin showed vacuolar cytoplasmic changes of epidermal cells mainly in the samples of xenografts irradiated by the lowest doses of gamma and electron beam radiation. The staining with orcein revealed damage of fine elastic fibers in the xenograft dermis at the dose of 25 kGy of both radiation types. Disintegration of epithelial basement membrane, especially in the xenografts, was induced by the dose of 15 kGy of electron beam radiation. The silver impregnation disclosed nuclear chromatin condensation mainly in human amnion at the lowest doses of both radiation types and disintegration of the fine collagen fibers in the papillary dermis induced by the lowest dose of electron beam and by the higher doses of gamma radiation. Irradiation by both, gamma rays and the electron beam, causes similar changes on cells and extracellular matrix, with significant damage of the basement membrane and of the fine and elastic and collagen fibers in the papillary dermis, the last caused already by low dose electron beam radiation. PMID:26649556

  19. Contribution of aquaporin 9 and multidrug resistance-associated protein 2 to differential sensitivity to arsenite between primary cultured chorion and amnion cells prepared from human fetal membranes

    Energy Technology Data Exchange (ETDEWEB)

    Yoshino, Yuta [Department of Clinical Molecular Genetics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392 (Japan); Yuan, Bo, E-mail: [Department of Clinical Molecular Genetics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392 (Japan); Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, 1550 4th St, RH584E Box 2911 San Francisco, CA 94158-2911 (United States); Kaise, Toshikazu [Laboratory of Environmental Chemodynamics, School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392 (Japan); Takeichi, Makoto [Yoneyama Maternity Hospital, 2-12 Shin-machi, Hachioji, Tokyo 192-0065 (Japan); Tanaka, Sachiko; Hirano, Toshihiko [Department of Clinical Pharmacology, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392 (Japan); Kroetz, Deanna L. [Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, 1550 4th St, RH584E Box 2911 San Francisco, CA 94158-2911 (United States); Toyoda, Hiroo [Department of Clinical Molecular Genetics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392 (Japan)


    Arsenic trioxide (arsenite, As{sup III}) has shown a remarkable clinical efficacy, whereas its side effects are still a serious concern. Therefore, it is critical to understand the effects of As{sup III} on human-derived normal cells for revealing the mechanisms underlying these side effects. We examined the effects of As{sup III} on primary cultured chorion (C) and amnion (A) cells prepared from human fetal membranes. A significant dose-dependent As{sup III}-mediated cytotoxicity was observed in the C-cells accompanied with an increase of lactate dehydrogenase (LDH) release. Higher concentrations of As{sup III} were required for the A-cells to show cytotoxicity and LDH release, suggesting that the C-cells were more sensitive to As{sup III} than the A-cells. The expression levels of aquaporin 9 (AQP9) were approximately 2 times higher in the C-cells than those in the A-cells. Both intracellular arsenic accumulation and its cytotoxicity in the C-cells were significantly abrogated by sorbitol, a competitive AQP9 inhibitor, in a dose-dependent manner. The protein expression levels of multidrug resistance-associated protein (MRP) 2 were downregulated by As{sup III} in the C-cells, but not in the A-cells. No significant differences in the expression levels of MRP1 were observed between C- and A-cells. The protein expression of P-glycoprotein (P-gp) was hardly detected in both cells, although a detectable amount of its mRNA was observed. Cyclosporine A, a broad-spectrum inhibitor for ABC transporters, and MK571, a MRP inhibitor, but not PGP-4008, a P-gp specific inhibitor, potently sensitized both cells to As{sup III}-mediated cytotoxicity. These results suggest that AQP9 and MRP2 are involved in controlling arsenic accumulation in these normal cells, which then contribute to differential sensitivity to As{sup III} cytotoxicity between these cells. -- Highlights: Black-Right-Pointing-Pointer Examination of effect of As{sup III} on primary cultured chorion (C) and amnion

  20. [Comparison of the effects of taurine and magnesium on electrical characteristics of artificial and natural membranes. V. Study on the human amnion of the antagonism between magnesium, taurine and polluting metals]. (United States)

    Bara, M; Guiet-Bara, A; Durlach, J


    The effects of metal pollutants (Pb, Cd, Hg, As) were studied on strips of human amnion isolated from the placental zone put in between two Ussing chambers with Hanks' solution at 37 degrees C and pH 7.4. The total conductance Gt through the human isolated amnion was decreased on the fetal side by Pb and As; on the maternal side by Cd, Hg and As. When Gt was decreased by metal pollutants, Mg or taurine (TA) were added in the external medium to induce an antagonism between Mg or TA and metal pollutants. The addition of Mg increased significantly the Gt reduced by Pb, Cd and Hg, but had no effect on the Gt reduced by As. The addition of taurine increased significantly the Gt reduced by Cd and Hg, but had no effect on the Gt reduced by Pb and As. Dixon's kinetics (Gt as a function of the Mg or TA concentration when the metal pollutant concentration increased) indicate that there is a competitive inhibition between Mg-Pb and Mg-Cd (the inhibition constant Ki is lower with Pb (= 2.5) than with Cd (= 11.4) and suggests a greater antagonism between Mg-Pb than between Mg-Cd). Moreover, there appears to be a noncompetitive inhibition between Mg-Hg, TA-Cd and TA-Hg. These results indicate that Mg and TA, on the fetal side, exert an action on the same sites and that, on the maternal side, their action takes place on the same sites and also on different ones. Also, TA can be considered as a partial magnesium agonist, at least in the human amnion.

  1. A prospective, randomised comparative study of weekly versus biweekly application of dehydrated human amnion/chorion membrane allograft in the management of diabetic foot ulcers (United States)

    Zelen, Charles M; Serena, Thomas E; Snyder, Robert J


    The aim of this study is to determine if weekly application of dehydrated human amnion/chorion membrane allograft reduce time to heal more effectively than biweekly application for treatment of diabetic foot ulcers. This was an institutional review board-approved, registered, prospective, randomised, comparative, non-blinded, single-centre clinical trial. Patients with non-infected ulcers of ≥ 4 weeks duration were included for the study. They were randomised to receive weekly or biweekly application of allograft in addition to a non-adherent, moist dressing with compressive wrapping. All wounds were offloaded. The primary study outcome was mean time to healing. Overall, during the 12-week study period, 92·5% (37/40) ulcers completely healed. Mean time to complete healing was 4·1 ± 2·9 versus 2·4 ± 1·8 weeks (P = 0·039) in the biweekly versus weekly groups, respectively. Complete healing occurred in 50% versus 90% by 4 weeks in the biweekly and weekly groups, respectively (P = 0·014). Number of grafts applied to healed wounds was similar at 2·4 ± 1·5 and 2·3 ± 1·8 for biweekly versus weekly groups, respectively (P = 0·841). These results validate previous studies showing that the allograft is an effective treatment for diabetic ulcers and show that wounds treated with weekly application heal more rapidly than with biweekly application. More rapid healing may decrease clinical operational costs and prevent long-term medical complications. PMID:24618401

  2. Stem cell therapy to protect and repair the developing brain: a review of mechanisms of action of cord blood and amnion epithelial derived cells. (United States)

    Castillo-Melendez, Margie; Yawno, Tamara; Jenkin, Graham; Miller, Suzanne L


    In the research, clinical, and wider community there is great interest in the use of stem cells to reduce the progression, or indeed repair brain injury. Perinatal brain injury may result from acute or chronic insults sustained during fetal development, during the process of birth, or in the newborn period. The most readily identifiable outcome of perinatal brain injury is cerebral palsy, however, this is just one consequence in a spectrum of mild to severe neurological deficits. As we review, there are now clinical trials taking place worldwide targeting cerebral palsy with stem cell therapies. It will likely be many years before strong evidence-based results emerge from these trials. With such trials underway, it is both appropriate and timely to address the physiological basis for the efficacy of stem-like cells in preventing damage to, or regenerating, the newborn brain. Appropriate experimental animal models are best placed to deliver this information. Cell availability, the potential for immunological rejection, ethical, and logistical considerations, together with the propensity for native cells to form teratomas, make it unlikely that embryonic or fetal stem cells will be practical. Fortunately, these issues do not pertain to the use of human amnion epithelial cells (hAECs), or umbilical cord blood (UCB) stem cells that are readily and economically obtained from the placenta and umbilical cord discarded at birth. These cells have the potential for transplantation to the newborn where brain injury is diagnosed or even suspected. We will explore the novel characteristics of hAECs and undifferentiated UCB cells, as well as UCB-derived endothelial progenitor cells (EPCs) and mesenchymal stem cells (MSCs), and how immunomodulation and anti-inflammatory properties are principal mechanisms of action that are common to these cells, and which in turn may ameliorate the cerebral hypoxia and inflammation that are final pathways in the pathogenesis of perinatal brain

  3. Contribution of aquaporin 9 and multidrug resistance-associated protein 2 to differential sensitivity to arsenite between primary cultured chorion and amnion cells prepared from human fetal membranes. (United States)

    Yoshino, Yuta; Yuan, Bo; Kaise, Toshikazu; Takeichi, Makoto; Tanaka, Sachiko; Hirano, Toshihiko; Kroetz, Deanna L; Toyoda, Hiroo


    Arsenic trioxide (arsenite, As(III)) has shown a remarkable clinical efficacy, whereas its side effects are still a serious concern. Therefore, it is critical to understand the effects of As(III) on human-derived normal cells for revealing the mechanisms underlying these side effects. We examined the effects of As(III) on primary cultured chorion (C) and amnion (A) cells prepared from human fetal membranes. A significant dose-dependent As(III)-mediated cytotoxicity was observed in the C-cells accompanied with an increase of lactate dehydrogenase (LDH) release. Higher concentrations of As(III) were required for the A-cells to show cytotoxicity and LDH release, suggesting that the C-cells were more sensitive to As(III) than the A-cells. The expression levels of aquaporin 9 (AQP9) were approximately 2 times higher in the C-cells than those in the A-cells. Both intracellular arsenic accumulation and its cytotoxicity in the C-cells were significantly abrogated by sorbitol, a competitive AQP9 inhibitor, in a dose-dependent manner. The protein expression levels of multidrug resistance-associated protein (MRP) 2 were downregulated by As(III) in the C-cells, but not in the A-cells. No significant differences in the expression levels of MRP1 were observed between C- and A-cells. The protein expression of P-glycoprotein (P-gp) was hardly detected in both cells, although a detectable amount of its mRNA was observed. Cyclosporine A, a broad-spectrum inhibitor for ABC transporters, and MK571, a MRP inhibitor, but not PGP-4008, a P-gp specific inhibitor, potently sensitized both cells to As(III)-mediated cytotoxicity. These results suggest that AQP9 and MRP2 are involved in controlling arsenic accumulation in these normal cells, which then contribute to differential sensitivity to As(III) cytotoxicity between these cells.

  4. Stem cell therapy to protect and repair the developing brain: a review of mechanisms of action of cord blood and amnion epithelial derived cells

    Directory of Open Access Journals (Sweden)

    Margie eCastillo-Melendez


    Full Text Available In the research, clinical and wider community there is great interest in the use of stem cells to reduce the progression, or indeed repair brain injury. Perinatal brain injury may result from acute or chronic insults sustained during fetal development, during the process of birth, or in the newborn period. The most readily identifiable outcome of perinatal brain injury is cerebral palsy, however this is just one consequence in a spectrum of mild to severe neurological deficits. As we review, there are now clinical trials taking place worldwide targeting cerebral palsy with stem cell therapies. It will likely be many years before strong evidence-based results emerge from these trials. With such trials underway, it is both appropriate and timely to address the physiological basis for the efficacy of stem-like cells in preventing damage to, or regenerating, the newborn brain. Appropriate experimental animal models are best placed to deliver this information. Cell availability, the potential for immunological rejection, ethical and logistical considerations, together with the propensity for native cells to form terratomas, make it unlikely that embryonic or fetal stem cells will be practical. Fortunately, these issues do not pertain to the use of human amnion epithelial cells (hAECs, or umbilical cord blood (UCB stem cells that are readily and economically obtained from the placenta and umbilical cord discarded at birth. These cells have the potential for transplantation to the newborn where brain injury is diagnosed or even suspected. We will explore the novel characteristics of hAECs and undifferentiated UCB cells, as well as UCB-derived endothelial progenitor cells and mesenchymal stem cells, and how immunomodulation and anti-inflammatory properties are principal mechanisms of action that are common to these cells, and which in turn may ameliorate the cerebral hypoxia and inflammation that are final pathways in the pathogenesis of perinatal brain

  5. Neuronal-like differentiation of single versus multiple treatments with human amnion-derived mesenchymal stem cells induced by basic fibroblast growth factor

    Institute of Scientific and Technical Information of China (English)

    Hongliang Jiao; Fangxia Guan; Xiang Hu; Jianbin Li; Hong Shan; Wei Li; Jun Li; Ying Du; Bo Yang; Yunfan Zhou


    BACKGROUND: Cultures from multiple portions of umbilical cord blood mesenchymal stem cells have been shown to undergo more rapid proliferation and attachment than single portions. OBJECTIVE: To observe growth of basic fibroblast growth factor (bFGF)-induced cultures of human amnion-derived mesenchymal stem cells (AMSCs) and differentiation into neuronal-like cells. DESIGN, TIME AND SETTING: Comparative observation. The study was performed at the Laboratory of Microbiology and Immunology, Basic Medical School of Zhengzhou University from January to May 2008.METHODS: Amnia from full-term, uterine-incision delivery were donated by 12 healthy women. AMSCs were obtained by cell separation and culture techniques, and were passaged and induced by bFGF. From the third passage, a total of 1 mL AMSCs, at a density of 1.0 ×10 4/mL, was separately harvested from six samples, which served as group A. A total of 1 mL AMSCs, at a density of 1.0×10 4 /mL, was harvested separately from the remaining six samples, which served as group B. A total of 0.5 mL from the six samples of group A and 0.5 mL from the six samples of group B were combined to form group C. MAIN OUTCOME MEASURES: Differences in cell quantity among the three groups were compared by cell quantification and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT)analysis. Expression of a glial cell marker, neuron-specific enolase, and nestin was detected in the three groups by immunocytochemistry. RESULTS: Cell quantification and MTT analysis of live cells, as well as AMSC absorbance, were significantly greater in group C compared with groups A and B at 18 days of culture (P<0.05), and no significant difference was observed between groups A and B. Glial fibrillary acidic protein, neuron-specific enolase, and nestin were expressed in all groups following bFGF induction. CONCLUSION: Mixed AMSC cultures promoted proliferation, and bFGF-induced AMSCs differentiated into neuronal-like cells.

  6. Comprehensive two-dimensional gel protein databases offer a global approach to the analysis of human cells: the transformed amnion cells (AMA) master database and its link to genome DNA sequence data

    DEFF Research Database (Denmark)

    Celis, J E; Gesser, B; Rasmussen, H H;


    A total of 3430 polypeptides (2592 cellular; 838 secreted) from transformed human amnion cells (AMA) labeled with [35S]methionine were separated and recorded using computer-aided two-dimensional (2-D) gel electrophoresis. A master 2-D gel database of cellular protein information that includes both......, mitochondria, Golgi, ribosomes, intermediate filaments, microfilaments and microtubules), levels in fetal human tissues, partial protein sequences (containing information on 48 human proteins microsequenced so far), cell cycle-regulated proteins, proteins sensitive to interferons alpha, beta, and gamma, heat......-specific proteins, to microsequence them and store the information in the database, to identify the corresponding genes, to search for homology with previously characterized proteins and to study the function of groups of proteins (pathways, organelles, etc.) that exhibit interesting regulatory properties...

  7. Amnion s and radio-sterilized porcine skin use as potential matrices for the development of human skin substitutes; Uso de amnios y piel porcina radioesterilizados como matrices potenciales para el desarrollo de sustitutos de piel humana

    Energy Technology Data Exchange (ETDEWEB)

    Martinez P, M. E.; Reyes F, M. L.; Reboyo B, D. [ININ, Carretera Mexico-Toluca s/n, 52750 Ocoyoacac, Estado de Mexico (Mexico); Velasquillo M, M. C.; Sanchez S, R.; Brena M, A. M.; Ibarra P, J. C., E-mail: [Instituto Nacional de Rehabilitacion, Calz. Mexico-Xochimilco No. 289, Col. Arenal de Guadalupe, 14389 Mexico D. F. (Mexico)


    The injuries by burns constitute a primordial problem of public health; they cause a high mortality index, severe physical and psychological disability, etc. The autologous skin transplant is the replacement therapy recommended for its treatment, but in patients that present a high percentage of burnt skin; this is not possible to carry out. Another strategy is the transplant of donated skin; however, due to the little donation that exists in our country is not very feasible to apply this treatment. A challenge of the tissues engineering is to develop biological skin substitutes, based on cells and amnion s, favoring the cutaneous regeneration and quick repair of injuries, diminishing this way the hospitalization expenses. At present skin substitutes that can equal to the same skin do not exist. On the other hand, the mesenchymal stromal cells (Msc) represent an alternative to achieve this objective; since has been demonstrated that the Msc participate in the tissue repair by means of inhibition of pro-inflammatory cytokines and differentiation to dermal fibroblasts and keratinocytes. To apply the Msc in cutaneous injuries a support material is required that to allow transplanting these cells to a lesion or burn. The radio-sterilized human amnion and the radio-sterilized porcine skin, processed by the Radio-Sterilized Tissues Bank of the Instituto Nacional de Investigaciones Nucleares (ININ), are biomaterials that are used as temporary cutaneous coverings. We suppose that these two matrices will be appropriate for the growth and maintenance in cultivation of the Msc, to generate two biological skin substitutes, in collaboration with the Biotechnology Laboratory of the Instituto Nacional de Rehabilitacion. (Author)

  8. Ex vivo expansion of hematopoietic stem- and progenitor cells from cord blood in coculture with mesenchymal stroma cells from amnion, chorion, Wharton's jelly, amniotic fluid, cord blood, and bone marrow. (United States)

    Klein, Caroline; Strobel, Julian; Zingsem, Jürgen; Richter, Richard H; Goecke, Tamme W; Beckmann, Matthias W; Eckstein, Reinhold; Weisbach, Volker


    In most cases, the amount of hematopoietic stem and progenitor cells (HSPCs) in a single cord blood (CB) unit is not sufficient for allogenic transplantation of adults. Therefore, two CB units are usually required. The ex vivo expansion of HSPCs from CB in coculture with mesenchymal stroma cells (MSCs) might be an alternative. It was investigated, whether bone marrow-derived MSCs, which have to be obtained in an invasive procedure, introduce a further donor and increases the risk of transmissible infectious diseases for the patient can be replaced by MSCs from amnion, chorion, Wharton's jelly, amniotic fluid, and CB, which can be isolated from placental tissue which is readily available when CB is sampled. In a two-step ex vivo coculture mononuclear cells from cryopreserved CB were cultured with different MSC-feederlayers in a medium supplemented with cytokines (stem cell factor, thrombopoietin [TPO], and granulocyte colony-stimulating factor). Expansion rates were analyzed as well, by long-term culture-initiating cell (LTC-IC) and colony-forming unit (CFU) assays, as by measuring CD34(+)- and CD45(+)-cells. Due to the comparably low number of 5×10(2) to 1×10(4) CD34(+)-cells per cm(2) MSC-monolayer, we observed comparably high expansion rates from 80 to 391,000 for CFU, 70 to 313,000 for CD34(+)-, and 200 to 352,000 for CD45(+)-cells. Expansion of LTC-IC was partly observed. Compared to the literature, we found a better expansion rate of CD34(+)-cells with MSCs from all different sources. This is probably due to the comparably low number of 5×10(2) to 1×10 CD34(+)-cells per cm(2) MSC-monolayer we used. Comparably, high expansion rates were observed from 80 to 391,000 for CFUs, 70 to 313,000 for CD34(+)-, and 200 to 352,000 for CD45(+)-cells. However, the expansion of CD34(+)-cells was significantly more effective with MSCs from bone marrow compared to MSCs from amnion, chorion, and Wharton's jelly. The comparison of MSCs from bone marrow with MSCs from CB and

  9. Effect of double-layer human amnion on wound healing in deep second-degree burns of animals%复层人羊膜覆盖治疗深Ⅱ度烫伤创面的动物实验

    Institute of Scientific and Technical Information of China (English)

    宋炬赫; 樊昌东; 李瑶


    背景:己证实单层人羊膜覆盖对浅度烧伤创面有一定的治疗作用,但仍存在很多不足.目的:观察自制复层辐射人羊膜覆盖治疗对鼠深Ⅱ度烫伤创面的效果.设计、时间及地点:随机分组,自身及组间对照动物实验,于2006-12/2007-05在哈尔滨医科大学第一临床医学院动物实验中心进行.材料:成年清洁级Wistar大鼠50只,随机分为伤后4,6,8,10,12 d组5组,每组10只.复层辐射人羊膜为自制,将新鲜人羊膜去除绒毛膜后,将羊膜层干燥,制成无孔及有孔的复层薄膜,再经Y射线辐射后塑膜包装.方法;所有动物背部脱毛,麻醉后浸入沸水中8 s致背部烫伤,制备深Ⅱ度烫伤模型.将每组创面积2等分,分别贴敷复层辐射人羊膜和油纱进行对照观察.主要观察指标:在伤后4,6,8,10,12 d取材,观察创面大体愈合情况,光镜、扫描电镜、透射电镜观察创面组织学变化,并进行创面细菌学检查.结果:50只大鼠全部进入结果分析.①贴敷油纱的创面:可见坏死物和痂皮,伤后12 d时肉眼仍见灶性小溃疡,光镜下见生真皮层少量毛囊、毛根、毛发再生,扫描电镜下见创面中心为坏死结痂,周边见少量毛发,大量炎性渗出物及白细胞存在,透射电镜显示细胞结构模糊不清.细菌学检查结果显示刨面细菌生长明显.②贴敷复层辐射人羊膜创面:伤后10,12d时创面愈合,肉眼见毛发规则丛生,和正常皮肤无区别,光镜、扫插电镜、透射电镜观察均显示伤后12 d时表皮各层均为正常皮肤组织.细菌学检查结果显示创面无细菌生长.结论:复层辐射人羊膜能能有效地保护鼠烫伤创面,促进创面的上皮生长,避免创面感染.缩短鼠深Ⅱ度烫伤创面的愈合时间.%BACKGROUND: Previous research reveals that, single-layer human amnion is effective to cure superficial bums, but it is still unsatisfactory.OBJECTIVE: To observe the effects of double-layer radiation

  10. Experimental study on the effect of human amnion on MMP-9 expression during healing process of rat skin incised wound%人羊膜对大鼠皮肤切创愈合中MMP-9表达影响的实验研究

    Institute of Scientific and Technical Information of China (English)

    罗秋燕; 边文玲; 申素芳


    目的:研究人羊膜对大鼠皮肤切创愈合过程中感染率的影响及基质金属蛋白酶-9(Matrix metalloproteinase-9,MMP-9)表达影响的研究及意义,探讨其促进皮肤愈合的机制,为其在临床的应用提供实验依据.方法:建立大鼠皮肤切创模型,观察切口愈合时间、感染率;在各时相点处死取材,应用免疫组织化学技术,通过计算机图像分析系统检测人胎羊膜对不同损伤时间大鼠皮肤组织中MMP -9的表达.结果:新鲜羊膜组感染率(6.7%)与对照组(33.3%)比较,创面感染率明显低于对照组,差异有统计学意义(P<0.05);在伤后12 h、1天、2天、3天,新鲜羊膜组MMP -9的表达高于对照组,差异有统计学意义(P<0.05),5天、7天两组的表达差异无统计学意义(P>0.05).结论:羊膜贴附创口能减少创口的感染率;在大鼠皮肤切创愈合的早期阶段,可通过促进创缘周围MMP -9的表达,加速清创,促进创口的愈合.%Objective: To research the effect of human amnion on infection rale and matrix metalloproteinase - 9 (MMP-9) expression during healing process of rat skin incised wound and its significance, explore the mechanism of its promoting effect on healing of rat skin, provide an experimental basis for the clinical application. Methods: The models of rat skin incised wound were established, the wound healing time and infection rate were observed; the rats were killed at different time points; immunohistochemical method was used to detect the effect of human amnion on MMP-9 expression in skin tissue at different time points by computer image analysis system. Results: The wound infection rate in fresh amnion group was 6.7% , which was significantly lower than that in control group (33. 3% ) (P 0.05 ) . Conclusion: Amnion adhered to wound can reduce the infection rate of incised wound; at early stage of rat skin incised wound healing, amnion can promote wound debridement and healing by promoting the expression of

  11. The mixed cultures and neuronal-like differentiation of human amnion-derived mesenchymal stem cells%人羊膜间充质干细胞混合培养及向神经样细胞分化

    Institute of Scientific and Technical Information of China (English)

    焦红亮; 王晓宁; 孙剑瑞; 李建斌; 关方霞; 杨波


    目的 观察羊膜间充质干细胞(HAMs)可诱导分化为神经样细胞混合培养的生长以及经碱性成纤维细胞生长因子(bFGF)诱导HAMs向神经样细胞分化.方法 采用细胞分离和培养技术获取HAMs,分离HAMs后传代培养,加入bFGF诱导分化.当细胞传至第3代时,随机取培养的6份HAMs(半量)为A组,6份HAMs(半量)为B组,将A、B组各剩余的半量HAMs混合为C组.3组细胞密度均为1.0 x 107/L.以活细胞计数和噻唑蓝(MTT)比色法比较3组细胞扩增数量,免疫组织化学法检测HAMs胶质纤维酸性蛋白(GFAP)、神经元特异性烯醇化酶(NSE)、巢蛋白(Nestin)的表达.以活细胞计数和MTT比色法比较3组细胞扩增数量,免疫组织化学检测HAMs神经胶质细胞标志物、神经元特异性烯醇化酶和巢蛋白的表达.结果 活细胞计数结果显示,第18天A组HAMs为(4.80±1.49)×104个,B组HAMs为(5.00±1.75) xl04个,C组HAMs为(8.90±3.46)×104个.MTT比色检测结果均显示,第18天A组HAMs为0.242±3.408,B组HAMs为0.245±5.226,C组HAMs为0.321 ±1.321(P <0.05).经bFGF诱导后均表达GFAP 、NSE和Nestin.结论 混合HAMs之间有互相促增殖作用,HAMs具有较强的可塑性,经bFGF诱导的HAMs可表达GFAP、NSE和Nestin.%Objective To observe growth of basic fibroblast growth factor (bFGF)-induced cultures of human amnion-derived mesenchymal stem cells (HAMs) and differentiation into neuronal-like cells.Comparative observation.Methods Amnia from full-term,uterine-incision delivery were donated by 12 healthy women.HAMs were obtained by cell separation and culture techniques,and were passaged and induced by bFGF.From the third passage,a total of 1 ml HAMs,at a density of 1.0 × 107/L,was separately harvested from six samples,which served as group A.A total of 1 ml HAMs,at a density of 1.0 × 107/L,was harvested separately from the remaining six samples,which served as group B.A total of 0.5 ml from the six samples of group A and 0.5 ml from the

  12. Transplantation of human amnion-derived mesenchymal stem cells alleviates ischemia-reperfusion-induced acute lung injury after cardiopulmonary bypass%人羊膜间充质干细胞移植减轻体外循环再灌注肺损伤

    Institute of Scientific and Technical Information of China (English)

    强勇; 梁贵友; 余丽梅; 齐斌; 高振宇


    BACKGROUND:In recent years, mesenchymal stem cels exhibit a good prospect in organ or tissue repair and therefore, and therefore, cel transplantation based on mesenchymal stem cel plasticity can promote cel regeneration and functional recovery from lung injury after cardiopulmonary bypass. OBJECTIVE:To investigate the effects of human amnion-derived mesenchymal stem cels (hAMSCs) transplantation on ischemia-reperfusion-induced acute lung injury in dogs after cardiopulmonary bypassand its mechanism for regulating inflammatory cytokines. METHODS:Eighteen adult healthy mongrel dogs were randomly divided into three groups (n=6 per group): black group (cardiopulmonary bypass with 1 mL physiological saline injectionvia the femoral vein without blocking the aorta), control group (cardiopulmonary bypass with blocking the aorta for 1 hour and then opening the aorta for 15 minutes plus 1 mL physiological saline injectionvia the femoral vein), experiment group (cardiopulmonary bypass with blocking the aorta for 1 hour and then opening the aorta for 15 minutes plus femoral vein injection of 1 mL physiological saline containing 2×107 hAMSCs). Arterial blood samples of 2 mL were taken to calculate oxygenation index and respiratory index before cardiopulmonary bypass (T1), 15 minutes (T2), 1 hour (T3), 2 hours (T4), 3 hours (T5) after opening the aorta. 8 mL intravenous blood samples were taken to detect the serum tumor necrosis factor α, matrix metaloproteinase-9, interleukin-8 and interleukin-10 by ELISA. Meanwhile, western blot assay was used to detect the expression of nuclear factor-κB in lung tissues, and histopathological changes of lung tissues observed under optical microscope. RESULTS AND CONCLUSION:Compared with the control group, the oxygenation index was significantly increased in the experimental group at 2 and 3 hours after transplantation, and the respiratory index was remarkably decreased at 1, 2, 3 hours after transplantation. Compared with the control group

  13. 人羊膜间充质干细胞培养上清液对人成纤维细胞生物学功能的影响%Effects of culture supernatant of human amnion mesenchymal stem cells on biological characteristics of human fibroblasts

    Institute of Scientific and Technical Information of China (English)

    吴骐而; 吕璐; 辛海明; 罗亮; 童亚林; 莫永亮; 岳毅刚


    Objective To investigate the effects of culture supernatant of human amnion mesenchymal stem cells (hAMSCs-CS) on biological characteristics of human fibroblasts.Methods (1) hAMSCs were isolated from deprecated human fresh amnion tissue of placenta and then sub-cultured.The morphology of hAMSCs on culture day 3 and hAMSCs of the third passage were observed with inverted phase contrast microscope.(2) Two batches of hAMSCs of the third passage were obtained,then the expression of vimentin of cells was observed with immunofluorescence method,and the expression of cell surface marker CD90,CD73,CD105,and CD45 was detected by flow cytometer.(3) hAMSCs-CS of the third passage at culture hour 72 were collected,and the content of insulin-like growth factor Ⅰ (IGF-Ⅰ),vascular endothelial growth factor (VEGF),epidermal growth factor (EGF),and basic fibroblast growth factor (bFGF) were detected by enzyme-linked immunosorbent assay.(4) Hunan fibroblasts were isolated from deprecated human fresh prepuce tissue of circumcision and then sub-cultured.Human fibroblasts of the third passage were used in the following experiments.Cells were divided into blank control group and 10%,30%,50%,and 70% hAMSCs-CS groups according to the random number table (the same grouping method below),with 48 wells in each group.Cells in blank control group were cultured with DMEM/F12 medium containing 2% fetal bovine serum (FBS),while cells in the latter 4 groups were cultured with DMEM/F12 medium containing corresponding volume fraction of hAMSCs-CS and 2% FBS.The proliferation activity of cells was detected by cell counting kit 8 and microplate reader at culture hour 12,24,48,and 72,respectively,and corresponding volume fraction of hAMSCs-CS which causing the best proliferation activity of human fibroblasts was used in the following experiments.(5) Human fibroblasts were divided into blank control group and 50% hAMSCs-CS group and treated as in (4),with 4 wells in each group,at post

  14. 人羊膜匀浆上清液对脂多糖致伤的大鼠肺微血管内皮细胞增殖及分泌炎症因子的影响%Effect of supernatant of human amnion homogenate on lipopolysaccharide induced pulmonary microvascular endotheli-al cells injury and their proliferation and expression of proinflammatory factors in rats

    Institute of Scientific and Technical Information of China (English)

    陈云鹏; 朱富军; 龚震宇; 辛海明; 王磊; 童亚林; 刘亮; 吕璐; 莫永亮; 詹球; 阳齐琼; 梁静


    长因子、细胞因子,对 LPS致伤的 RPMVECs增殖具有促进作用,并减少致伤后炎症因子分泌。%Objective:To investigate the protective effect of supernatant of human amnion homogenate (hAHS)on proliferation and expression of proinflammatory mediators by lipopolysaccharide (LPS)induced inj ured pulmonary microvascular endothelial cells of rats (RPMVECs).Methods:hAHS was prepared from fresh human amnion. The total protein content and the content of epithelial growth factor (EGF),basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF),interleukin-4 (IL-4),IL-10,angiogenin-1 (Ang-1),humanβ-defensin2 (HBD2)of hAHS were determined with Coomassie blue staining and ELISA. The effect of 0,10%,15%, 20%,25% hAHS on cell proliferation activity of RPMVECs was respectively determined with MTT assay,in or-der to determine the optimal concentration of hAHS on promoting RPMVECs proliferation. According to different co-culture conditions,RPMVECs were randomly divided into 4 groups:group N (cultured with 10%FBS+DMEM/F12),group A(10%FBS+DMEM/F12+15%hAHS),group B (10%FBS+DMEM/F12+LPS),and group C (10%FBS+DMEM/F12+15%hAHS+LPS). At 0,12,24,48,72 hours after culturing with the corre-sponding medium of each group,optical density values (A values)of each group were determined respectively with MTT assay to determine the proliferation activity,and the contents of IL-6,IL-8,TNF-αlevels in the culture su-pernates were also determined by ELISA at 6,8,10,12 and 24 hours. Results:The total protein concentration of hAHS was (725.125±12.625)mg/L,and levels of EGF,bFGF,VEGF,IL-4,IL-10,Ang-1,HDB2 were re-spectively(504.785±4.665)ng/L,(4.426±0.138)ng/L,(0.185±0.006)ng/L,(25.650±4.104)ng/L,(13.733 ±2.197)ng/L,(15.561±0.496)ng/L,(4.763±0.714)ng/L.10%-20% hAHS was shown to promote prolifer-ation of RPMVECs,and 15% hAHS,and the best result was observed on 7 and 9 days. The proliferation rate of RPMVECs in 25% hAHS group at 7,9 and 11 days was lower than those in the 0%hAHS group (P<0

  15. 羊膜卷填塞治疗角膜穿孔羊膜的演变及对角膜内皮细胞的影响%The evolution of amnion and influnce on corneal endothelium in amniotic roll plugging for corneal perforation

    Institute of Scientific and Technical Information of China (English)

    范军华; 何伟杰; 叶瑞珍; 李学喜


    Objective To observe the evolution of amnion and influnce on corneal endothelium in amniotic roll plugging combined with C3F8 tamponade in anterior chamber to treat corneal perforation.Methods 56 eyes of 56 patients with corneal perforation accepted the operation of amniotic roll plugging combined with C3F8 tamponade in anterior chamber and the concentration of C3F8 gas was 20%.Anterior chamber depth,aqueous leakage,bubbles maintenance time,corneal endothelial cell density and morphology and the fusion of amniotic roll and corneal tissue were observed.Results Corneal perforation of 56 eyes were all repaired without amniotic roll falling off.The fusion of amniotic roll and corneal tissue was observed at 2 months after operation,and the amniotic roll and corneal tissue fused well and no gap was examined.The C3F8 gas stayed in the anterior chamber for 5 ~ 16 days,average of (8.2 ± 4.8) days.There was no significant difference in corneal endothelial cell density between preoperative and postoperative 1 month or 2 month.The percentage of hexagonal cells decreased at postoperative 1 month compared with preoperative (t =2.13,P > 0.05),and there was no significant difference between postoperative 1 month and 2 months (t =1.65,P > 0.05).Conclusion Amniotic roll plugging combined with C3 F8 gas tamponade in anterior chamber can effectively repair corneal perforation,and the amniotic roll and cornea can fuse well after operation.C3F8 gas can stay in the anterior chamber long enough to maintain the anterior chamber depth and prevent leakage of aqueous humor.Little influnce on the corneal endothelium was observed.%目的 观察羊膜卷填塞联合C3F8气体填充治疗角膜穿孔术后羊膜的演变及其对角膜内皮细胞的影响.方法 因各种原因所致角膜穿孔而施行羊膜卷填塞联合前房C3F8气体填充术共56例(56眼).术中以20%C3F8气体填充前房.术后观察前房深度、房水渗漏情况、气泡维持时间、角膜内皮细

  16. 自体角膜缘干细胞移植与羊膜移植治疗翼状胬肉的对比研究%Comparative Study on treatment of pterygium with auto-corneallimbus stem cell transplantation and amnion transplantation

    Institute of Scientific and Technical Information of China (English)

    蔡春梅; 付湘媛; 张晶; 邓江稳


    目的 比较自体角膜缘干细胞移植、羊膜移植和单纯翼状胬肉切除三种手术方式治疗翼状胬肉的治愈及复发情况.设计回顾性病例系列.研究对象自2007年8月至2010年8月在解放军第二炮兵总医院眼科应用上述三种手术方法治疗并有随访记录的翼状胬肉患者193例(217眼).方法 根据手术方法的不同,将193例(217眼)翼状胬肉患者分为3组:角膜缘干细胞移植组(A组)73例81眼;羊膜移植组(B组)54例63眼;单纯翼状胬肉切除组(C组)66例73眼.术后平均随访1年(12.2±2.3)月,观察并比较三组患者的治疗效果、复发率和并发症情况.主要指标复发率和并发症.结果 随访期间,A组复发4眼,复发率为4.94%;B组复发10眼,复发率为15.9%;C组复发23眼,复发率为31.5%(A与B组、A与C组、B与C组相比,x2值分别为4.828、18.746、4.498,P值分别为0.028、0.000、0.034).随访期内,A组未发生睑球粘连,B组有1眼(1.59%)出现睑球粘连,C组有8眼(10.96%)出现睑球粘连.结论 与单纯翼状胬肉切除术和羊膜移植术相比,自体角膜缘干细胞移植治疗翼状胬肉并发症少、复发率低,是治疗翼状胬肉较好的手法方法.%Objective To compare the curative and recurrent situation for the treatment of pterygium among three operative methods (simple excision of prerygium,combined excision of pterygium with auto-corneallimbus stem cell transplantation, and combined excision of pterygium with amnion transplantation). Design Retrospective case series. Participants 193 cases (217 eyes )with pterygium,who underwent the operation in The Second Artillery General Hospital of PLA from Aug. 2007 to Aug. 2010. Methods 193 cases (217 eyes ) with pterygium were divided into 3 groups according to the operative method: combined excision of pterygium with auto-comedlimbus stem cell transplantation (group A )73 cases (81 eyes), combined excision of pterygium with amnjon transplantation (group B) 54 cases (63

  17. Human Amnion Epithelial Cells Induced to Express Functional Cystic Fibrosis Transmembrane Conductance Regulator


    Murphy, Sean V.; Rebecca Lim; Philip Heraud; Marian Cholewa; Mark Le Gros; de Jonge, Martin D.; Howard, Daryl L.; David Paterson; Courtney McDonald; Anthony Atala; Graham Jenkin; Wallace, Euan M


    Cystic fibrosis, an autosomal recessive disorder caused by a mutation in a gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR), remains a leading cause of childhood respiratory morbidity and mortality. The respiratory consequences of cystic fibrosis include the generation of thick, tenacious mucus that impairs lung clearance, predisposing the individual to repeated and persistent infections, progressive lung damage and shortened lifespan. Currently there is no cure fo...

  18. Morphological analysis of amnion stored in glycerol sterilized with different doses of ionizing radiation

    Energy Technology Data Exchange (ETDEWEB)

    Soares, Fernando Augusto N.; Santin, Stefany P.; Martino Junior, Antonio C.; Machado, Luci Diva B.; Mathor, Monica B., E-mail: [Instituto de Pesquisas Energeticas e Nucleares (CTR/IPEN/CNEN-SP), Sao Paulo, RJ (Brazil). Centro de Tecnologia das Radiacoes


    The amniotic membrane (AM) is the innermost layer of the fetal membranes (placenta), widely used in transplantation being a tissue that combines anti-inflammatory, antimicrobial and antifibrotic, and limited immunogenicity. The tissue can be used a bandage biological for treatment of burns and skin wounds, chronic ulcers, reconstructions from different body areas, including ophthalmic repairs. In the last decades the amniotic membrane has been used widely also as a carrier substrate to transfer tissues cultured 'in vitro'. The use of fresh membrane has some limitations, the main ones are being necessary your quick use and the inability to obtain full security for certain infections. Other types of preservation require a processing thereof. The radiosterilization is an alternative for ensuring quality and safety of tissues used in transplants, and other clinical applications in order to minimize the risk of contamination of the receptor tissue. Therefore, the objective of this study was to test various doses of radiation using two sources of ionizing radiation: the cobalt-60 irradiator Gamma and Electron Beam Accelerator (E.B.). A tissue analysis was done by visual and tactile qualitative analysis, semi-quantitative (solid colorimetry) and light microscopy to observe morphological and physic-chemical changes after the irradiation of AM preserved in glycerol, comparing the results obtained with the sample not irradiated. It was noted that at higher doses, for both radiation sources, irradiated membranes suffered greater color change, becoming yellowish and thereby reducing their initial malleability. (author)

  19. Surfactant Protein-A as an Anti-Inflammatory Component in the Amnion: Implications for Human Pregnancy1


    Lee, Deug-Chan; Romero, Roberto; Kim, Chong Jai; Chaiworapongsa, Tinnakorn; Tarca, Adi L.; Lee, JoonHo; Suh, Yeon-Lim; Mazaki-Tovi, Shali; Vaisbuch, Edi; Mittal, Pooja; Draghici, Sorin; Erez, Offer; Kusanovic, Juan Pedro; Hassan, Sonia S; Kim, Jung-Sun


    The mechanism of mouse parturition is thought to involve myometrial infiltration by amniotic fluid (AF) macrophages, activated by surfactant protein-A (SP-A). In humans, the concentration of AF SP-A decreases during labor, and no fetal macrophages are found in the myometrium after labor. Therefore, it appears that the mechanisms of labor in mice and humans are different. We investigated a potential role for SP-A in human pregnancy and parturition by examining SP-A expression patterns in AF an...

  20. Comparative study of effects of magnesium and taurine on electrical parameters of natural and artificial membranes. VII. Effects on cellular and paracellular ionic transfer through isolated human amnion. (United States)

    Bara, M; Guiet-Bara, A; Durlach, J


    The comparative effects of 2 mM magnesium and taurine on various components of the human transamniotic conductance, Gt, were observed. The use of both microelectrodes and metabolic inhibitors enables 10 components of Gt to be distinguished: six cellular components (Na-K ATPase, Na-H antiport, Na-K-2Cl cotransport and Na, K, Cl channels), one coupling component, and three paracellular components (Na, K, Cl). Mg increased all components of Gt while taurine only increased five of them (Na and K channels, coupling, Na and K paracellular conductance). A potentiometric effect of taurine on Mg2+ modified membrane, obtained on paracellular components, was not measured on cellular components. There was only a vicarious effect between Mg and taurine on the non-enzymatic cellular and paracellular transfer of Na and K.

  1. Count of Amnionic Fluid Lamellar Body to Predict the Maturity of Fetal Lung%羊水板层体计数预测胎肺成熟度

    Institute of Scientific and Technical Information of China (English)

    周容; 熊庆; 游永; 周淑; 徐蜀渝; 张静丽; 杜泽丽


    目的探讨羊水板层体计数(Lamellar body count, LBC)预测胎肺成熟度的价值. 方法采用库尔特Micro Diff II全自动血细胞分析仪对41例正常足月妊娠剖宫产时的羊水标本进行LBC的测定,并对同一标本采用薄板层析法测定了卵磷脂/鞘磷脂比值(Lecithin-sphingomyelin, L/S比值). 结果(1)正常足月妊娠时羊水LBC为(86±43)×103(范围72.5×103~99.4×103).(2)羊水LBC与L/S比值之间存在正相关,r=0.66,P=0.001. 结论羊水LBC是一种快速、准确地判断肺成熟度的方法.


    Institute of Scientific and Technical Information of China (English)

    邢鸣鸾; 陈加平; 王晓峰; 徐进; 徐立红


    大田软海绵酸(Okadaic acid,OA)是一种C38的长链脂肪酸,属聚醚类海洋毒素,是腹泻性贝毒(Diarrhetic shellfish poisoning,DSP)的主要成分,化学结构见下图。最初从大田软海绵(Halichondria Okadai)和隐瓜海绵(H.melanodocia)中分离得到,后发现实际上是由共生于上述两种海绵的利马原甲藻(Prorocentrum lima)所产生。

  3. 翼状胬肉切除术移植结膜或羊膜的复发率比较%The comparison of the recurrence rates after pterygium excision combined with conjunctiva and with amnion transplantation

    Institute of Scientific and Technical Information of China (English)

    曾仁攀; 梁小琼; 王国平


    Objective To compare the recurrence rates after pterygium excision combined with autologous conjunctiva and amniotic membrane transplantation .Methods One hundred and thirty eyes of one hundred and eight patients who underwent pterygium surgery were randomly divided into group A and group B. Patients in group A received pterygium excision combined with autologous conjunctiva transplantation, including 70 eyes of 58 patients; and patients in group B received pterygium excision combined with amniotic membrane transplantation , including 60 eyes of 50 patients.All patients were followed up for 1 year .The tear film stability ( breaking up time ) and recurrence rates were recorded and analyzed.Results One year after the surgery, 8 eyes (11.42%) in group A had recurrent pterygium , and 14 eyes (23.33%) in group B had recurrent pterygium .The difference in recurrence rate between the two groups was statistically significant ( P0.05).Conclusion Pterygium excision combined with autologous conjunctiva transplantation can make lower recurrence rate than pterygium excision with amniotic membrane transplantation .%目的:比较翼状胬肉切除联合自体结膜移植术与联合羊膜移植术后的复发率。方法翼状胬肉手术108例(130眼)随机分为A、B两组。 A组为自体结膜移植术组(58例,70眼)。 B组为羊膜移植术组(50例,60眼)。术后随访1年,以泪膜破裂时间及复发率作为分析统计指标。结果通过术后1年的跟踪随访,翼状胬肉切除联合自体结膜移植术组有8眼(11.42%)复发。联合羊膜移植术组有14眼(23.33%)复发。两组复发率比较,差异有统计学意义(P<0.05)。两组泪膜破裂时间之间的差异无统计学意义(P>0.05)。结论翼状胬肉切除联合自体结膜移植术治疗翼状胬肉能更有效的降低术后复发率。

  4. Radiosensitivity of angiogenic and mitogenic factors in human amniotic membrane

    International Nuclear Information System (INIS)

    Amniotic membrane as a temporary biological dressing remains as a beneficial and cost-effective means of treating burns in developing countries. This medical application is attributed mainly to placental structural and biochemical features that are important for maintaining proper embryonic development. Since fresh amnions are nevertheless for straightforward clinical use and for preservation, radiation-sterilization is been performed to improve the safety of this placental material. However, like any other sterilization method, gamma-radiation may induce physical and chemical changes that may influence the biological property of the material. Thus, the aim of this study is to compare the effects of various levels of radiation-sterilization protocols for human amnions on angiogenic (neovascularization) and epithelial-mitogenic activities, both of which are physiological processes fundamental to wound healing. Water-soluble extract of non-irradiated amnions demonstrates a strong stimulatory effect on both cell proliferation and angiogenesis. No change in biological activity is seen in amnions irradiated at 25 kGy, the sterilization dose used by the Philippine Nuclear Research Institute (PNRI) for the production of radiation-sterilized human amniotic membranes (RSHAM). However, it appears that amniotic angiogenic factors are more radiosensitive than its mitogenic components, evident from the depressed vascularization of the chorioallantoic membrane (CAM) exposed to 35 kGy-irradiated amnions. The dose of 35 kGy is at present the medical sterilization dose used at the Central Tissue Bank in Warsaw (Poland) for the preparation of their amnion allografts. (Authors)

  5. 人羊膜间充质干细胞静脉移植治疗阿尔茨海默病APP+转基因鼠的有效性及安全性评估%Efficacy and safety assessment of intravenous transplantation of human amnion membrane mesenchymal stem cells in treatment of APP+ transgenic mice with Alzheimer's disease

    Institute of Scientific and Technical Information of China (English)

    郭亚男; 李远; 关方霞; 李国栋; 李祥生; 杨波; 杜英; 胡祥; 胡炜; 焦红亮


    背景:目前阿尔茨海默病的病因不明,课题组前期实验发现人羊膜间充质干细胞静脉移植可促进阿尔茨海驮病APP+转基因鼠的学习、记忆能力改善.目的:进一步评估人羊膜间充质干细胞静脉移植治疗阿尔茨海默病APP+转基因鼠的有效性和安全性.设计、时间及地点:细胞学体内实验与动物对照观察,于2008-05/12在郑州大学生物工程系、郑州大学基础医学院及河南省中医药研究院完成.材料:清洁级C57BL/6系APP+转基因胎鼠10只,合笼繁育后得到子代小鼠200只,按其繁殖情况和PCR榆测结果,取29只11月龄小鼠,分为APP+对照组10只、APP+细胞移植组10只、APP+正常组9只.羊膜标本由郑州大学第一附属医院产科提供.方法:体外分离培养人羊膜间充质干细胞,传至第3代后调整浓度为1×109 L-1的单细胞悬液.APP+细胞移植组每只小鼠尾静脉注射人羊膜间充质干细胞悬液O.5 mL,APP+对照组注射等量生理盐水,APP+正常组不作任何处理.主要观察指标:采用Morris水迷宫测定移植前后小鼠学习和记忆功能.移植当天开始称小鼠体质量,持续3周.移植后解剖小鼠,收集全血并分离血清,进行12项肿瘤标记物检测及心、肝、肾功能的血液生化指标检测,对人羊膜间充质干细胞移植的安全性作综合评价.结果:移植后15d各组逃避潜伏期比较,APP-正常组<APP+细胞移植组<APP+对照组,APP+细胞移植组小鼠逃避潜伏期明显短于APP+对照组(P<0.05);与移植前比较,移植后15d各组小鼠穿越平台象限的次数均有所增加,且APP+细胞移植组小鼠在平台象限停留时间明显延长.移植后3周内,3组小鼠体质量增长趋势差异无显著性意义(P>0.05),12项肿瘤标记物,9项血清肝肾功能生化指标及10项血液学指标的表达均无明显差异(P>0.05). 结论:人羊膜间充质干细胞经尾静脉移植后,可明显改善阿尔茨海默病小鼠的学习、记忆能力,未见致死致瘤现象发生,心、肝、肾功能未受影响,安全可行.

  6. 足月妊娠羊水过少患者羊膜和胎盘的组织病理学及组织化学变化%Histopathology and Histochemistry Changes of Amnion and Placenta with Oligohydramnios in Full Term Pregnancy

    Institute of Scientific and Technical Information of China (English)



    Objective:To explore the histopathologic and histochemistic changes of amniotic membrane and placenta with oligohydramnios in term pregnancy.Method:Amniotic membrane and placenta with normal amount amniotic fluid and oligohydramnios 15 cases in each group.The martial undergone fixation with 10% formaldehyde and routine paraffin section, HE staining and AB-PAS reaction.The slices were observed under light microscope and some index of the morphomatry acquired.Result:The amniotic cells of oligohydramnios became shriek and vesicle, even stratified metaplasia, PAS granules were reduced in their cytoplasm.Syncytail knots increased.AB-PAS reaction became weaken because of glycogen granules in cytoplasm were rare, the basement membrane of the capillaries at the villi became vary thinker, all these changes point on aging and disfunction.Conclusion:The histopathologic changes and glycogen reduce in amniotic cells, and number of syncytail knot raise, thinker vasculosyncytial membrane and rare glycogen et al in placenta villi take part in the pathological and physiological process of the oligohydramnios.%目的:探讨足月妊娠羊水过少患者的羊膜和胎盘的组织化学反应变化。方法:取足月分娩正常量羊水和羊水过少患者的羊膜和胎盘各15例,标本经10%甲醛固定,常规石蜡制片,分别进行HE和阿利新蓝-多糖(AB-PAS)法染色,光镜观察并进行形态计量学分析。结果:羊水过少羊膜细胞出现固缩、空泡变甚至鳞状化生,胞质内PAS阳性颗粒减少;胎盘滋养层细胞PAS反应明显减弱阳性颗粒缺失,合体结节增多,绒毛中轴毛细血管基膜多增厚,血管合体膜增厚等。结论:羊膜上皮形变、老化,糖原缺乏、血管合体膜增厚、合体结节增多,参与羊水过少的病理生理过程。

  7. 不同传代倍数人羊膜来源的间充质干细胞成骨成脂分化平衡的研究%Research on balance between osteogenic and adipogenic differentiation of human amnion - derived mesenchymal stem cells of different passage numbers

    Institute of Scientific and Technical Information of China (English)

    陈霞; 尹晓娟


    Objective To compare proliferative activity, osteogenic and adipogenic differentiation capacity among different passage numbers of human amniotic mesenchymal stem cells( hAMSCs ). Methods The hAMSCs were isolated from human placental basal decidua hy collagenase and trypsinase digestion methods. The superficial molecular markers of these cells were detected hy flow cytometry. After serial passage , passage 3 ,5 ,7 ,9( P3 , P5 , P7 , P9 )were singled out for the assay of proliferative activity by MTT method and the findings were delineated as growth curve. Furthermore, these cells of above four passages were respectively induced by adding osteogenic and adipogenic inducers. 3 w later, intracellular lipid deposition was verified by oil red O staining and extracellular calciumphosphate precipitates were stained with alizarin red. The positive rate of osteogenic staining and adipogenic rate were compared among different passage numbers of hAMSCs. Results hAMSCs of P9 still showed a colony - like growth property , but their proliferative activity was down regulated. Intracellular lipid droplets decreased from P5( P < 0.01 ). and extracellular calciumphosphate did from P7( P< 0.01 ). Conclusion hAMSCs can keep growth characteristics of mesenchymal stem cells during serial passage. However,multipotential differentiation capacity is varied among different passage numbers.%目的 比较不同传代倍数的人羊膜来源的间充质干细胞(hAMSCs)的增殖活性以及成骨、成脂的分化能力.方法 采用胶原酶和胰蛋白酶法分离人胎盘羊膜来源的MSCs,测定分离细胞的表面分子标志.对第3、5、7、9代hAMSCs绘制细胞生长曲线,分别加入成骨和成脂诱导剂,诱导3 w后行茜素红和油红O染色,比较不同传代倍数的hAMSCs成骨染色阳性率和成脂率.结果 hAMSCs传代至第9代时仍呈克隆样生长,但细胞增殖活力有下降;在第5代后成骨分化能力下降(P<0.01),而传代至第7代后,成脂分化能力才有下降(P<0.01).结论 hAMSCs在连续传代中能保持MSCs的生长特性,但多向诱导分化能力在不同的传代倍数中有一定差别.

  8. Periostin as a Biomarker of the Amniotic Membrane

    Directory of Open Access Journals (Sweden)

    Mariya P. Dobreva


    Full Text Available Tracing the precise developmental origin of amnion and amnion-derived stem cells is still challenging and depends chiefly on analyzing powerful genetic model amniotes like mouse. Profound understanding of the fundamental differences in amnion development in both the disc-shaped primate and human embryo and the cup-shaped mouse embryo is pivotal in particular when sampling amniotic membrane from nonprimate species for isolating candidate amniotic stem cells. The availability of molecular marker genes that are specifically expressed in the amniotic membrane and not in other extraembryonic membranes would be instrumental to validate unequivocally the starting material under investigation. So far such amniotic markers have not been reported. We postulated that bone morphogenetic protein (BMP target genes are putative amniotic membrane markers mainly because deficiency in one of several components of the BMP signaling cascade in mice has been documented to result in defective development of the early amnion. Comparative gene expression analysis of acknowledged target genes for BMP in different extraembryonic tissues, combined with in situ hybridization, identified Periostin (Postn mRNA enrichment in amnion throughout gestation. In addition, we identify and propose a combination of markers as transcriptional signature for the different extraembryonic tissues in mouse.

  9. Concurrent conjunctivitis and placentitis in aborted bovine fetuses. (United States)

    Murray, R D


    Consistent histopathological lesions were found in 10 out of 136 aborted fetuses examined during a three year period, using a multi-disciplinary diagnostic investigation technique. Fetuses exhibited a generalized mononuclear inflammatory cell infiltration, accompanied by distinctive lesions of conjunctival hyperplasia and goblet cell formation, alveolitis, and necrotic placentitis. In two cases where amnion was also examined, a chronic amnionitis was present. No consistent laboratory findings could be related to these cases. The fetal and placental lesions described were similar to those associated with experimental inoculation of Ureaplasma diversum in pregnant cows, and with field isolations of the same organism in aborting cattle.

  10. Some Histories of Molecular Evolution: Amniote Phylogeny, Vertebrate Eye Lens Evolution, and the Prion Gene

    NARCIS (Netherlands)

    Rheede, T. van


    In this thesis, the principles of molecular evolution and phylogeny are introduced in Chapter 1, while the subsequent chapters deal with the three topics mentioned in the title. Part I: Birds, reptiles and mammals are Amniota, organisms that have an amnion during their embryonal development. Even th

  11. Anencephaly: MRI findings and pathogenetic theories

    International Nuclear Information System (INIS)

    We describe the MRI appearances of an anencephalic newborn who survived for 13 h; particularities of this case are male gender and the absence of other associated malformations. Moreover, we discuss the pathogenetic theories of anencephaly, correlating MRI findings with embryological data. An exencephaly-anencephaly sequence due to amnion rupture is hypothesized. (orig.)

  12. Constriction band syndrome occurring in the setting of in vitro fertilization and advanced maternal age


    Rinker, Brian; Vasconez, Henry C.


    The debate as to the pathogenesis of constriction band syndrome began with Hippocrates and continues today. The exogenous theory attributes the condition to entanglement of the fetus in the amniotic remnants following premature rupture of the amnion, which is in contrast to the endogenous, or genetic, mechanism.

  13. Potential antitumor therapeutic strategies of human amniotic membrane and amniotic fluid-derived stem cells. (United States)

    Kang, N-H; Hwang, K-A; Kim, S U; Kim, Y-B; Hyun, S-H; Jeung, E-B; Choi, K-C


    As stem cells are capable of self-renewal and can generate differentiated progenies for organ development, they are considered as potential source for regenerative medicine and tissue replacement after injury or disease. Along with this capacity, stem cells have the therapeutic potential for treating human diseases including cancers. According to the origins, stem cells are broadly classified into two types: embryonic stem cells (ESCs) and adult stem cells. In terms of differentiation potential, ESCs are pluripotent and adult stem cells are multipotent. Amnion, which is a membranous sac that contains the fetus and amniotic fluid and functions in protecting the developing embryo during gestation, is another stem cell source. Amnion-derived stem cells are classified as human amniotic membrane-derived epithelial stem cells, human amniotic membrane-derived mesenchymal stem cells and human amniotic fluid-derived stem cells. They are in an intermediate stage between pluripotent ESCs and lineage-restricted adult stem cells, non-tumorigenic, and contribute to low immunogenicity and anti-inflammation. Furthermore, they are easily available and do not cause any controversial issues in their recovery and applications. Not only are amnion-derived stem cells applicable in regenerative medicine, they have anticancer capacity. In non-engineered stem cells transplantation strategies, amnion-derived stem cells effectively target the tumor and suppressed the tumor growth by expressing cytotoxic cytokines. Additionally, they also have a potential as novel delivery vehicles transferring therapeutic genes to the cancer formation sites in gene-directed enzyme/prodrug combination therapy. Owing to their own advantageous properties, amnion-derived stem cells are emerging as a new candidate in anticancer therapy.

  14. Distribution of CA 125 in placental tissues. (United States)

    Fuith, L C; Müller-Holzner, E; Marth, C; Perkmann, E; Zeimet, A; Daxenbichler, G


    The presence of the tumor marker CA 125 was studied in different compartments of the human placenta. Levels of CA 125 in the cytosol of chorionic villi ranged from 27-17100 U/g (median 560 U/g). In the placental amnion and chorion concentrations ranged from 175-29000 U/g, median 1060 U/g and were not statistically different. In the umbilical cord values were significantly lower (range 44-7600 U/g; median 180 U/g). Maternal serum probes were above the upper limit of normal in all cases (range 48-500 U/ml; median 131 U/ml). Immunohistochemistry detected CA 125 exclusively within the amniotic cells of the placenta and the umbilical cord. This might be because CA 125 fixes more to insoluble structures in the amnion or because of contamination of chorionic villi with the underlying decidua.

  15. Invasiveness and Ploidy of Human Mammary Carcinomas in Short-Term Culture (United States)

    Smith, Helene S.; Liotta, Lance A.; Hancock, Miriam C.; Wolman, Sandra R.; Hackett, Adeline J.


    Invasiveness and ploidy were examined in cultures of human epithelial cells derived from nonmalignant breast tissue, primary breast carcinomas, and breast cancer effusion metastases. Successful short-term culture was achieved from approximately 70% of the primary breast cancers. These primary cancers were essentially diploid by flow cytometry and karyotype in contrast to the effusion metastases, which were mostly aneuploid. The diploid tumor cells retained their malignant phenotype in culture as demonstrated by invasion into a denuded human amnion basement membrane. In contrast, epithelial cells cultured from nonmalignant mammary tissue did not invade the amnion. We suggest that the diploid carcinoma cultures may be useful for investigating the essential differences between normal and malignant cells and may complement information derived from studies of tumor cell lines with grossly aberrant karyotypes.

  16. Velamentous Cord Insertion in a Singleton Pregnancy: An Obscure Cause of Emergency Cesarean—A Case Report


    Juliana Rocha; Joana Carvalho; Fernanda de Costa; Isabel Meireles; Olímpia do Carmo


    Approximately 2% of low-risk pregnant women still require an emergency Cesarean section after the onset of labor. Because it is likely that half of these cases are associated with placental and umbilical cord abnormalities, it is thought that prenatal detection of such abnormalities would reduce the number of emergency cesarean sections in low-risk women. Velamentous cord insertion is an abnormal cord insertion in which the umbilical vessels diverge as they traverse between the amnion and cho...

  17. Case report of non-healing surgical wound treated with dehydrated human amniotic membrane


    Riordan, Neil H; George, Ben A; Chandler, Troy B; McKenna, Randall W


    Introduction Non-healing wounds can pose a medical challenge as in the case of vasculopathic venostasis resulting in a surgical ulcer. When traditional approaches to wound care fail, an amniotic patch (a dehydrated tissue allograft derived from human amnion) can function as a biologic scaffold to facilitate and enhance tissue regeneration and rehabilitation. Background Amniotic AlphaPatches contain concentrated molecules of PGE2, WNT4, and GDF-11 which have angiogenic, trophic, and anti-infla...

  18. Left-sided gastroschisis with placenta findings: case report and literature review


    Shi, Yang; Farinelli, Christine K; Chang, Michael S; CARPENTER, PHILIP M.


    Gastroschisis is a congenital abdominal-wall defect that typically occurs to the right of the umbilicus. Only twenty-one cases of left-sided gastroschisis have been described in the literature. Here we report a large left-sided gastroschisis with pulmonary hypoplasia, scoliosis, ventricular septal defect and absence of gallbladder. Section of placental membranes revealed vacuolization of the amnion, without increased macrophage infiltration of the chorion. Postmortem comparative genomic hybri...

  19. The Amniotic Membrane: Development and Potential Applications - A Review. (United States)

    Favaron, P O; Carvalho, R C; Borghesi, J; Anunciação, A R A; Miglino, M A


    Foetal membranes are essential tissues for embryonic development, playing important roles related to protection, breathing, nutrition and excretion. The amnion is the innermost extraembryonic membrane, which surrounds the foetus, forming an amniotic sac that contains the amniotic fluid (AF). In recent years, the amniotic membrane has emerged as a potential tool for clinical applications and has been primarily used in medicine in order to stimulate the healing of skin and corneal diseases. It has also been used in vaginal reconstructive surgery, repair of abdominal hernia, prevention of surgical adhesions and pericardium closure. More recently, it has been used in regenerative medicine because the amniotic-derived stem cells as well as AF-derived cells exhibit cellular plasticity, angiogenic, cytoprotective, immunosuppressive properties, antitumoural potential and the ability to generate induced pluripotent stem cells. These features make them a promising source of stem cells for cell therapy and tissue engineering. In this review, we discussed the development of the amnion, AF and amniotic cavity in different species, as well as the applicability of stem cells from the amnion and AF in cellular therapy.

  20. Transdifferentiation of human amniotic epithelial cells into acinar cells using a double-chamber system. (United States)

    Huang, Gui-Lin; Zhang, Ni-Ni; Wang, Jun-Sheng; Yao, Li; Zhao, Yu-Jie; Wang, Yu-Ying


    This study investigated the transdifferentiation of stem cells from human amnion tissue into functional acinar cells (ACs) using a co-culture system. Human amniotic epithelial cells (hAECs) were isolated from amnion tissue by mechanical mincing and enzymatic digestion. After primary culture, the phenotype of the cells was identified by flow cytometry (FCM) and immunocytochemical staining. hAECs were co-cultured with submandibular gland acinar cells of SD rats using a double-chamber system. The expression of α-amylase was determined by immunocytochemical method and fluorescent real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) after induction for 1 and 2 weeks, respectively. Digestion with trypsin is an effective method for isolating hAECs from amnion tissue. These cells were positive for CD29 and CK19 and weakly positive for CD44 and α-amylase. Within 2 weeks, α-amylase in hAECs increased with induction time. The expression of α-amylase in hAECs was increased 3.38-fold after co-culturing for 1 week. This ratio increased to 6.6-fold, and these cells were positive for mucins, after co-culturing for 2 weeks. hAECs possess the potential to differentiate into ACs in vitro. They might be a stem cell resource for clinical applications of cell replacement therapy in salivary gland dysfunction diseases. PMID:22800093

  1. Clinical application of amniotic membranes on a patient with epidermolysis bullosa. (United States)

    Martínez Pardo, M E; Reyes Frías, M L; Ramos Durón, L E; Gutiérrez Salgado, E; Gómez, J C; Marín, M A; Luna Zaragoza, D


    The case of a patient with dystrophic epidermolysis bullosa treated with radiosterilised amniotic membranes is presented. The disorder is a congenital disease characterised by a poor desmosomal junction in the keratinocyte membrane. After proper donor screening, amnios were collected at Hospital Central Sur de Alta Especialidad (HCSAE), PEMEX and microbiological analysis was performed at Universidad Nacional Autónoma de México, FQUNAM, (Biology Dept. of the Chemistry Faculty, National Autonomous University of Mexico), before and after radiation sterilisation. Processing, packaging and sterilisation were performed at Instituto Nacional de Investigaciones Nucleares, ININ, (National Nuclear Research Institute). The patient, a ten-year-old boy with severe malnutrition, extensive loss of skin and pseudomonad infection in the whole body, was treated with gentle debridement in a Hubbard bath. Later amnion application was performed with sterilised amnios by using two different processes, in one of which the amnion was sterilised with paracetic acid, preserved in glycerol, kindly donated by the German Institute for Tissue and Cell Replacement and applied by Dr. Johannes C. Bruck, IAEA visiting expert, and the other amnion was processed at ININ: air dried and sterilised by gamma radiation at dose of 30 kGy. After spontaneous epithelisation was successfully promoted for seven days, the pain was alleviated and mobility was improved in a few hours and the patient's general condition was so improved that in a month he was discharged. Unfortunately, because this disease is revertive and has malignant degeneration, the prognosis is not good. PMID:10853786

  2. Biomechanical assays amniotic membrane preserved in glycerol correlating with optical coherence tomography (OCT) and thermal gravimetric analysis (TG)

    Energy Technology Data Exchange (ETDEWEB)

    Soares, Fernando Augusto N.; Santin, Stefany P.; Martino Junior, Antonio C.; Machado, Luci Diva B.; Freitas, Anderson Z.; Mathor, Monica B., E-mail: [Instituto de Pesquisas Energetias Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)


    Amnion or amniotic membranes (AM) are interchangeable terms used in the literature being internal part of the fetal membranes, non-vascular and multicellular tissue. The amnion has been widely used as a graft ophthalmic surgical as well as carrier substrate stem cell tissue equivalent for ocular surface reconstruction. The AM reduces scar formation and inflammation on the ocular surface, promotes epithelization also been used as a biological bandage covering the wound or burns, reducing dehydration and allowing regeneration of these areas. The amnion has usually 0.02 to 0.5 mm thick and consists of five subsequent layers: epithelium, basement membrane, compact layer, fibroblast layer and spongy layer. The mechanical strength from the membrane structure as well as the elasticity are factors attractive to the use of amnion as a surgical graft. Higher levels of rigidity and strength may improve the graft resistance necessary to resist the stress induced during growth of the new tissue formed. The amniotic membrane is obtained at elective caesarean section and subsequently, under sterile conditions, sectioned and separated from chorion and placenta, and free blood clots. The serological tests are done at the time of collection of tissue and 6 months after delivery to confirm the results. There are different methods for storing MA in tissue banks as fresh, high concentrations of glycerol, among others. The use of fresh membrane has some limitations due to the need to rapid use and high risk of contamination, however the amniotic membrane in glycerol has antiviral and antibacterial property which is dependent on the concentration, time and temperature. The AM used in transplants must be sterile to prevent the transmission of any disease. Although sterilization by radiation is an effective procedure, it can interfere on the membrane structure. Thus, verification of potential changes caused by ionizing radiation in amnion was made using the tensile test by calculating the

  3. The cytoplasmic 60 kDa progesterone receptor isoform predominates in the human amniochorion and placenta at term

    Directory of Open Access Journals (Sweden)

    Bell Stephen C


    Full Text Available Abstract Background The mechanism that initiates human parturition has been proposed to be 'functional progesterone withdrawal' whereby the 116 kDa B-isoform of the progesterone receptor (PR-B switches in favour of the 94 kDa A-isoform (PR-A in reproductive tissues. Recently, other PR isoforms, PR-S, PR-C and PR-M generated from the same gene have been identified and partially characterised. Methods and Results Using immunohistochemical, western blotting and RT-PCR techniques, evidence is provided that indicates the major PR isoform present in human term fetal membranes (amnion and chorion and syncytiotrophoblast of the placenta is neither of the classical nuclear PR-B or PR-A isoforms but is the N-terminally truncated 60 kDa PR-C isoform. Evidence is also provided that this 60 kDa isoform resides in the cytoplasm of the expressing cell types. Data are also presented to show that PR-B, PR-A and PR-S isoforms are essentially absent from the amnion and chorion, whereas PR isoforms A, B, C and S are all present in the decidua, with PR-A being the major isoform. The syncytiotrophoblast of the placenta contains the cytoplasmic 60 kDa isoform, but not isoforms PR-A, PR-B or PR-S. Conclusion The major PR isoform in the amnion, chorion and placenta is a 60 kDa protein that could be PR-C, suggesting that the cytoplasmic isoform has a specific role in extra-embryonic tissues and may be involved in the regulation of human parturition.

  4. Type VII collagen associated with the basement membrane of amniotic epithelium forms giant anchoring rivets which penetrate a massive lamina reticularis. (United States)

    Ockleford, C D; McCracken, S A; Rimmington, L A; Hubbard, A R D; Bright, N A; Cockcroft, N; Jefferson, T B; Waldron, E; d'Lacey, C


    In human amnion a simple cuboidal epithelium and underlying fibroblast layer are separated by an almost acellular compact layer rich in collagen types I and III. This (>10 μm) layer, which may be a thick lamina reticularis, apparently presents an unusual set of conditions. Integration of the multilaminous tissue across it is apparently achieved by waisted structures which we have observed with the light microscope in frozen, paraffin-wax and semi-thin resin sections. We have also captured transmission and scanning electron micrographs of the structures. These structures which cross the compact layer we call "rivets". The composition of these "rivets" has been examined immunocytochemically and in three dimensions using the confocal laser scanning epi-fluorescence microscope. The rivets contain type VII collagen and an α6 integrin. They associate with type IV collagen containing structures (basement membrane lamina densa and spongy coils) and a special population of fibroblasts which may generate, maintain or anchor rivets to the underlying mesenchymal layer. Although type VII collagen is well known to anchor basal lamina to underlying mesodermal collagen fibres these "rivets" are an order of magnitude larger than any previously described type VII collagen containing anchoring structures. Intriguing possible functions of these features include nodes for growth of fibrous collagen sheets and sites of possible enzymatic degradation during regulated amnion weakening approaching term. If these sites are confirmed to be involved in amnion degradation and growth they may represent important targets for therapeutic agents that are designed to delay preterm premature rupture of the membranes a major cause of fetal morbidity and mortality.

  5. Inflammatory Response of Human Gestational Membranes to Ureaplasma parvum Using a Novel Dual-Chamber Tissue Explant System. (United States)

    Potts, Lauren C; Feng, Liping; Seed, Patrick C; Jayes, Friederike L; Kuchibhatla, Maragatha; Antczak, Brian; Nazzal, Matthew K; Murtha, Amy P


    Preterm premature rupture of membranes (PPROM) is often associated with intra-amniotic inflammation and infection. Current understanding of the pathogenesis of PPROM includes activation of pro-inflammatory cytokines and proteolytic enzymes leading to compromise of membrane integrity. The impact of exposure to bacterial pathogens, including Ureaplasma parvum, on gestational membranes is poorly understood. Our objective was to develop a dual-chamber system to characterize the inflammatory response of gestational membranes to U. parvum in a directional nature. Full-thickness human gestational membrane explants, with either choriodecidua or amnion oriented superiorly, were suspended between two washers in a cylindrical device, creating two distinct compartments. Brilliant green dye was introduced into the top chamber to assess the integrity of the system. Tissue viability was evaluated after 72 h using a colorimetric cell proliferation assay. Choriodecidua or amnion was exposed to three doses of U. parvum and incubated for 24 h. Following treatment, media from each compartment were used for quantification of U. parvum (quantitative PCR), interleukin (IL)-8 (enzyme-linked immunosorbent assay), and matrix metalloproteinase (MMP)-2 and MMP-9 activity (zymography). We observed that system integrity and explant viability were maintained over 72 h. Dose-dependent increases in recovered U. parvum, IL-8 concentration, and MMP-2 activity were detected in both compartments. Significant differences in IL-8 concentration and MMP-9 activity were found between the choriodecidua and amnion. This tissue explant system can be used to investigate the inflammatory consequences of directional bacterial exposure for gestational membranes and provides insight into the pathogenesis of PPROM and infectious complications of pregnancy. PMID:27009041

  6. Cat amniotic membrane multipotent cells are nontumorigenic and are safe for use in cell transplantation

    Directory of Open Access Journals (Sweden)

    Vidane AS


    Full Text Available Atanasio S Vidane,1 Aline F Souza,1 Rafael V Sampaio,1 Fabiana F Bressan,2 Naira C Pieri,1 Daniele S Martins,2 Flavio V Meirelles,2 Maria A Miglino,1 Carlos E Ambrósio2 1Department of Surgery, Faculty of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil; 2Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of São Paulo, São Paulo, Brazil Abstract: Amnion-derived mesenchymal stem cells (AMSCs are multipotent cells with an enhanced ability to differentiate into multiple lineages. AMSCs can be acquired through noninvasive methods, and therefore are exempt from the typical ethical issues surrounding stem cell use. The objective of this study was to isolate and characterize AMSCs from a cat amniotic membrane for future application in regenerative medicine. The cat AMSCs were harvested after mechanical and enzymatic digestion of amnion. In culture medium, the cat AMSCs adhered to a plastic culture dish and displayed a fibroblast-like morphology. Immunophenotyping assays were positive for the mesenchymal stem cell-specific markers CD73 and CD90 but not the hematopoietic markers CD34, CD45, and CD79. Under appropriate conditions, the cat AMSCs differentiated into osteogenic, chondrogenic, and adipogenic cell lineages. One advantage of cat AMSCs was nonteratogenicity, assessed 4 weeks post injection of undifferentiated AMSCs into immunodeficient mice. These findings suggest that cat amniotic membranes may be an important and useful source of mesenchymal stem cells for clinical applications, especially for cell or tissue replacement in chronic and degenerative diseases. Keywords: amnion, cats, cell differentiation, fetal membranes, mesenchymal cells

  7. Progesterone, Inflammatory Cytokine (TNF-α), and Oxidative Stress (H2O2) Regulate Progesterone Receptor Membrane Component 1 Expression in Fetal Membrane Cells. (United States)

    Meng, Yan; Murtha, Amy P; Feng, Liping


    Progesterone receptor membrane component 1 (PGRMC1) is an important novel mediator of progesterone (P4) function in fetal membrane cells. We demonstrated previously that PGRMC1 is differentially expressed in fetal membranes among pregnancy subjects and diminished in preterm premature rupture of membrane subjects. In the current study, we aim to elucidate whether PGRMC1 expression is regulated by P4, tumor necrosis factor α (TNF-α), and H2O2 in fetal membrane cells. Primary cultured membrane cells were serum starved for 24 hours followed by treatments of P4, 17 hydroxyprogesterone caproate, and medroxyprogesterone 17 acetate (MPA) at 10(-7) mol/L with ethanol as vehicle control; TNF-α at 10, 20, and 50 ng/mL with phosphate-buffered saline (PBS) as control; and H2O2 at 10 and 100 μmol/L with culture media as control for 24, 48, and 72 hours. The messenger RNA (mRNA) and protein expression of PGRMC1 was quantified using polymerase chain reaction and Western blotting, respectively. We found that PGRMC1 protein expression was regulated by MPA, TNF-α, and H2O2 in a dose-dependent manner. This regulation is also specific to the type of cell (amnion, chorion, or decidua). The upregulation of PGRMC1 by MPA might be mediated through glucocorticoid receptor (GR) demonstrated using amnion and chorion cells model with GR knockdown by specific small interfering RNA transfection. The mRNA expression of PGRMC1 was decreased by H2O2 (100 μmol/L) treatment in amnion cells, which might ultimately result in downregulation of PGRMC1 protein as our data demonstrated. None of other treatments changed PGRMC1 mRNA level in these cells. We conclude that these stimuli act as regulatory factors of PGRMC1 in a cell-specific manner. PMID:26919974

  8. Cat amniotic membrane multipotent cells are nontumorigenic and are safe for use in cell transplantation


    Vidane AS; Souza AF; Sampaio RV; Bressan FF; Pieri NC; Martins DS; Meirelles FV; Miglino MA; Ambrósio CE


    Atanasio S Vidane,1 Aline F Souza,1 Rafael V Sampaio,1 Fabiana F Bressan,2 Naira C Pieri,1 Daniele S Martins,2 Flavio V Meirelles,2 Maria A Miglino,1 Carlos E Ambrósio2 1Department of Surgery, Faculty of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil; 2Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of São Paulo, São Paulo, Brazil Abstract: Amnion-derived ...

  9. Risk of infertility following fetography and amniofetography

    International Nuclear Information System (INIS)

    297 women were subjected to prenatal diagnosis, in 220 of them amniocentesis was performed to start an amnion cell culture and 77 women (26%) were diagnosed by fetography and amniofetography, respectively. Following intra-amniotic injection of contrast medium abortion occured in 9% and premature delivery in 28.6% of the probands. The perinatal mortality was 13%. After amniocentesis the abortion rate was 3.2% and premature delivery took place in 4.5% of the patients. The perinatal mortality was but slightly increased (2.3%). The differences were statistically secured. Following fetography and amniofetography prophylactic cerclage is recommended

  10. Evolution of the Radio-sterilized Tissues Bank of the Instituto Nacional de Investigaciones Nucleares; Evolucion del Banco de Tejidos Radioesterilizados del Instituto Nacional de Investigaciones Nucleares

    Energy Technology Data Exchange (ETDEWEB)

    Martinez P, M. E.; Reyes F, M. L.; Luna Z, D., E-mail: [ININ, Carretera Mexico-Toluca s/n, 52750 Ocoyoacac, Estado de Mexico (Mexico)


    The Radio-sterilized Tissues Bank (RTB) of the Instituto Nacional de Investigaciones Nucleares (ININ) was established in 1997, as national project supported by the IAEA, who provided equipment, training and expert missions. In July of 1999, the Secretaria de Salud (Mexico) granted the sanitary license to the RTB. The first radio-sterilized tissue was the amnion. Later on, the process of frozen and lyophilized pig skin was development. Both tissue types are used as biological dressings in patient with burns, ulcers or on injuries difficult to heal, the amnion is also used for the damage treatment in the ocular surface. In 2003, the ININ improved the facilities of the RTB and in August of that same year the certification of the Quality Administration System was obtained under the standard ISO 9001:2000, at the present time ISO 9001:2008. The support of the IAEA granted to Mexico was of 1997-2004 and 2009-2012, by means of regional and inter regional projects where was obtained equipment and training mainly, for personal of the ININ and other institutions. With the last project supported by the IAEA, two very important documents were generated for the tissues bank of Argentina, Brazil, Colombia, Costa Rica, Cuba, Ecuador, El Salvador, Mexico, Peru, Uruguay and Venezuela, strengthening the interaction among these countries. To offer more radio-sterilized tissues to the medical community, in June of 2007 the RTB signed and agreement with the Secretaria de Salud of the Mexico State by means of the Centro Estatal de Transplantes del Estado de Mexico, for the process of muscle-skeletal tissue, skin and amnion. At present, besides amnion and pig skin, in the RTB there is in existence powdered and chips bone; concluding the validation stage the human skin process is. As a social function of the ININ, the RTB has contributed to the health improvement of patients of more than 50 hospital institutions, mainly those that assist to patient of low resources in different cities of

  11. Cloning of a cDNA encoding a novel human nuclear phosphoprotein belonging to the WD-40 family

    DEFF Research Database (Denmark)

    Honoré, B; Leffers, H; Madsen, Peder;


    -134]. The protein contains a nuclear targeting signal (KKKGK), and fractionation of transformed human amnion cells (AMA) in karyoplasts and cytoplasts confirmed that it is predominantly localized in the nucleus. Database searching indicated that IEF SSP 9502 is a putative human homologue of the Saccharomyces......We have cloned and expressed in vaccinia virus a cDNA encoding an ubiquitous 501-amino-acid (aa) phosphoprotein that corresponds to protein IEF SSP 9502 (79,400 Da, pI 4.5) in the master 2-D-gel keratinocyte protein database [Celis et al., Electrophoresis 14 (1993) 1091-1198]. The deduced aa...

  12. Expression of immunoreactive urocortin in human tissue

    Institute of Scientific and Technical Information of China (English)

    GU Qing; Vicki L Clifton; CUI Ying; HUI Ning; ZHOU Xiao-ning; HE Qian; HAN Qing-feng; SHA Jin-yan; Roger Smith


    To localize where urocortin is expressed in human tissue in an attempt to study its physiological functions. Methods: Expression of immunoreactive urocortin in different human tissue was examined using a specific urocortin antibody and the immunoperoxidase staining method. Results: Immunoreactive urocortin was observed in the anterior pituitary cells, decidual stromal cells, syncytiotrophoblasts, amnion epithelium, the vascular smooth muscles of myometrium, fallopian tube and small intestine. Conclusion: The study indicates that urocortin is expressed in some specific areas of human tissue. The data are consistent with the hypothesis that urocortin is produced locally as an endocrine factor, which may act as a neural regulator and a regulator of local blood flow.

  13. Membrane Vesicles of Group B Streptococcus Disrupt Feto-Maternal Barrier Leading to Preterm Birth. (United States)

    Surve, Manalee Vishnu; Anil, Anjali; Kamath, Kshama Ganesh; Bhutda, Smita; Sthanam, Lakshmi Kavitha; Pradhan, Arpan; Srivastava, Rohit; Basu, Bhakti; Dutta, Suryendu; Sen, Shamik; Modi, Deepak; Banerjee, Anirban


    Infection of the genitourinary tract with Group B Streptococcus (GBS), an opportunistic gram positive pathogen, is associated with premature rupture of amniotic membrane and preterm birth. In this work, we demonstrate that GBS produces membrane vesicles (MVs) in a serotype independent manner. These MVs are loaded with virulence factors including extracellular matrix degrading proteases and pore forming toxins. Mice chorio-decidual membranes challenged with MVs ex vivo resulted in extensive collagen degradation leading to loss of stiffness and mechanical weakening. MVs when instilled vaginally are capable of anterograde transport in mouse reproductive tract. Intra-amniotic injections of GBS MVs in mice led to upregulation of pro-inflammatory cytokines and inflammation mimicking features of chorio-amnionitis; it also led to apoptosis in the chorio-decidual tissue. Instillation of MVs in the amniotic sac also resulted in intrauterine fetal death and preterm delivery. Our findings suggest that GBS MVs can independently orchestrate events at the feto-maternal interface causing chorio-amnionitis and membrane damage leading to preterm birth or fetal death. PMID:27583406

  14. The potential of mesenchymal stem cells derived from amniotic membrane and amniotic fluid for neuronal regenerative therapy. (United States)

    Kim, Eun Young; Lee, Kyung-Bon; Kim, Min Kyu


    The mesenchymal stem cells (MSCs), which are derived from the mesoderm, are considered as a readily available source for tissue engineering. They have multipotent differentiation capacity and can be differentiated into various cell types. Many studies have demonstrated that the MSCs identified from amniotic membrane (AM-MSCs) and amniotic fluid (AF-MSCs) are shows advantages for many reasons, including the possibility of noninvasive isolation, multipotency, self-renewal, low immunogenicity, anti-inflammatory and nontumorigenicity properties, and minimal ethical problem. The AF-MSCs and AM-MSCs may be appropriate sources of mesenchymal stem cells for regenerative medicine, as an alternative to embryonic stem cells (ESCs). Recently, regenerative treatments such as tissue engineering and cell transplantation have shown potential in clinical applications for degenerative diseases. Therefore, amnion and MSCs derived from amnion can be applied to cell therapy in neuro-degeneration diseases. In this review, we will describe the potential of AM-MSCs and AF-MSCs, with particular focus on cures for neuronal degenerative diseases.

  15. Tissue-specific Differentiation Potency of Mesenchymal Stromal Cells from Perinatal Tissues. (United States)

    Kwon, Ahlm; Kim, Yonggoo; Kim, Myungshin; Kim, Jiyeon; Choi, Hayoung; Jekarl, Dong Wook; Lee, Seungok; Kim, Jung Min; Shin, Jong-Chul; Park, In Yang


    Human perinatal tissue is an abundant source of mesenchymal stromal cells(MSCs) and lacks the ethical concerns. Perinatal MSCs can be obtained from various tissues as like amnion, chorion, and umbilical cord. Still, little is known of the distinct nature of each MSC type. In this study, we successfully isolated and cultured MSCs from amnion(AMSCs), chorion(CMSCs), and umbilical cord(UC-MSCs). Proliferation potential was different among them, that AMSCs revealed the lowest proliferation rate due to increased Annexin V and senescence-associated β-galactosidase positive cells. We demonstrated distinct characteristic gene expression according to the source of the original tissue using microarray. In particular, genes associated with apoptosis and senescence including CDKN2A were up-regulated in AMSCs. In CMSCs, genes associated with heart morphogenesis and blood circulation including HTR2B were up-regulated. Genes associated with neurological system processes including NPY were up-regulated in UC-MSCs. Quantitative RT-PCR confirmed the gene expression data. And in vitro differentiation of MSCs demonstrated that CMSCs and UC-MSCs had a more pronounced ability to differentiate into cardiomyocyte and neural cells, respectively. This study firstly demonstrated the innate tissue-specific differentiation potency of perinatal MSCs which can be helpful in choosing more adequate cell sources for better outcome in a specific disease.

  16. Radioactive labeling of proteins in cultured postimplantation mouse embryos. I. Influence of the embryo preparation method

    International Nuclear Information System (INIS)

    Conditions for optimum incorporation of radioactive amino acids into proteins of cultured postimplantation mouse embryos were investigated under the aspect of using these proteins for two-dimensional electrophoretic separations followed by fluorography. The aim was to obtain highly radioactive proteins under conditions as physiological as possible. Embryos at Days 10, 11, and 12 of gestation were prepared in different ways and incubated for 4 h in Tyrode's solution containing [3H]amino acids (mixture) at a concentration of 27 microCi/ml medium. The preparations were: (a) yolk sac opened, placenta and blood circulation intact; (b) yolk sac and amnion opened, placenta and blood circulation intact (Day 10 embryos only); (c) placenta, yolk sac, and amnion removed (embryo naked); (d) naked embryos cut randomly into pieces (Day 10 embryos only). After incubation whole embryos or certain parts (tail, liver, rest body) were investigated by determining the radioactivity taken up by the protein. The results are given in dpm per mg protein per embryo. Radioactivity of proteins was about 3 times higher in naked embryos than in embryos left in their yolk sacs. This was true for all three stages investigated. However, the degree of radioactivity in the various parts of naked embryos differed by a factor of 15, whereas radioactivity was evenly distributed in embryos incubated in their yolk sacs. Therefore, embryos prepared according to the first method (see above) fulfilled the conditions required at the best

  17. Membrane Vesicles of Group B Streptococcus Disrupt Feto-Maternal Barrier Leading to Preterm Birth (United States)

    Sthanam, Lakshmi Kavitha; Srivastava, Rohit; Basu, Bhakti; Dutta, Suryendu; Sen, Shamik; Modi, Deepak


    Infection of the genitourinary tract with Group B Streptococcus (GBS), an opportunistic gram positive pathogen, is associated with premature rupture of amniotic membrane and preterm birth. In this work, we demonstrate that GBS produces membrane vesicles (MVs) in a serotype independent manner. These MVs are loaded with virulence factors including extracellular matrix degrading proteases and pore forming toxins. Mice chorio-decidual membranes challenged with MVs ex vivo resulted in extensive collagen degradation leading to loss of stiffness and mechanical weakening. MVs when instilled vaginally are capable of anterograde transport in mouse reproductive tract. Intra-amniotic injections of GBS MVs in mice led to upregulation of pro-inflammatory cytokines and inflammation mimicking features of chorio-amnionitis; it also led to apoptosis in the chorio-decidual tissue. Instillation of MVs in the amniotic sac also resulted in intrauterine fetal death and preterm delivery. Our findings suggest that GBS MVs can independently orchestrate events at the feto-maternal interface causing chorio-amnionitis and membrane damage leading to preterm birth or fetal death. PMID:27583406

  18. Puerperal endometritis after abdominal twin delivery. (United States)

    Suonio, S; Huttunen, M


    The infectious complications of 122 consecutive abdominal twin deliveries over the period 1984-1989 were analyzed in a prospective clinical study, comparing them with 761 singleton abdominal deliveries over the period 1984-1986. The incidence of endometritis was nearly three-fold after twin deliveries and the incidence of abdominal wound infections nearly two-fold compared with singleton abdominal pregnancies (13.1/4.7% and 5.6/3.0%). The risk of amnionitis was increased ten-fold, 6 hours after rupture of the membranes in abdominal twin delivery, but no connection was found between amnionitis and endometritis, as in singleton abdominal deliveries. Multiple regression analysis indicated only two risk factors as regards puerperal endometritis after abdominal twin delivery: age under 25 years (odds ratio 6.9, 95% confidence limits 1.9-24.8), an association also seen in singleton abdominal deliveries, and a period of more than 6 hours from rupture of membranes to delivery (odds ratio 7.8, 95% confidence limits 2.1-28.5). Multiple pregnancy appears to be associated with an increased risk of endometritis. The etiological factors remain unknown, but a large placental bed and/or immunological factors may be implicated. PMID:8160537

  19. High plasticity in epithelial morphogenesis during insect dorsal closure

    Directory of Open Access Journals (Sweden)

    Kristen A. Panfilio


    Insect embryos complete the outer form of the body via dorsal closure (DC of the epidermal flanks, replacing the transient extraembryonic (EE tissue. Cell shape changes and morphogenetic behavior are well characterized for DC in Drosophila, but these data represent a single species with a secondarily reduced EE component (the amnioserosa that is not representative across the insects. Here, we examine DC in the red flour beetle, Tribolium castaneum, providing the first detailed, functional analysis of DC in an insect with complete EE tissues (distinct amnion and serosa. Surprisingly, we find that differences between Drosophila and Tribolium DC are not restricted to the EE tissue, but also encompass the dorsal epidermis, which differs in cellular architecture and method of final closure (zippering. We then experimentally manipulated EE tissue complement via RNAi for Tc-zen1, allowing us to eliminate the serosa and still examine viable DC in a system with a single EE tissue (the amnion. We find that the EE domain is particularly plastic in morphogenetic behavior and tissue structure. In contrast, embryonic features and overall kinetics are robust to Tc-zen1RNAi manipulation in Tribolium and conserved with a more distantly related insect, but remain substantially different from Drosophila. Although correct DC is essential, plasticity and regulative, compensatory capacity have permitted DC to evolve within the insects. Thus, DC does not represent a strong developmental constraint on the nature of EE development, a property that may have contributed to the reduction of the EE component in the fly lineage.

  20. 羊膜移植及单纯胬肉切除治疗复发性翼状胬肉的疗效比较%Clinical observations the transplantation ofanmion and the pure removal of ptarygiurrl to treat the recurrent pterygium

    Institute of Scientific and Technical Information of China (English)



    Objective:The effect analyses by comparing and observing the transplantation of fresh amnion and the pure removal of pterygium to treat the recurrent pterygium.Method:71 cases ofrecurren Pterygium (74 eyes)is divides into2 groups (named A,B) randomly.Group A (with 38 cases,39 eyes)is operated by removing the recurrent pterygium and the transplantation of the fresh amnion.GroupB (with 33 cases.35 eyes)m only operated by removing the pteryginm.After operation,visits lasting 6~36 months are followed and comparations about the pafients’symptoms,the wound matches and the pterygium relapses among the 2 groups are carried out.Result In group A,12 eyes relapsed from the whole 39 eyes,and the rate of relapsing is 30.8%.In group B,30 eyes relapsed from the whole 35 eyes.with the rate of relapsing 85.5%.Conclusion:Compared with the effect ofthe pure removal pterygium to treat recurrent pterygium, there exist lower rates of relapsing in corneal limbastemcellautografting and thetransplantation of fresh amnion.In addition. and it is the effective option in treating recurrent pterygium.%目的:比较新鲜羊膜移植与单纯胬肉切除治疗复发性翼状胬肉的疗效。方法:对复发性翼状胬肉71例(74只眼),随机分为A、B两组,A组38例,39只眼,行复发性翼状胬肉切除联合新鲜羊膜移植术。B组33例,35只眼,行单纯胬肉切除。手术后随访6~36个月。比较两组患者的症状、创伤愈合情况、胬肉复发情况。结果:A组39只眼中复发12只眼,复发率30.8%。 B组35只眼中复发30例,复发率85.5%。结论:新鲜羊膜移植较单纯胬肉切除明显降低复发性翼状胬肉手术后复发率。

  1. Antioxidative defense enzymes in placenta protect placenta and fetus in inherited thrombophilia from hydrogen peroxide


    Jelena Bogdanovic Pristov; Ivan Spasojevic; Željko Mikovic; Vesna Mandic; Nikola Cerovic; Mihajlo Spasic


    Our aim was to investigate the activities of antioxidative defense enzymes in the placenta, fetal blood and amnion fluid in inherited thrombophilia. Thrombophilia was associated with nearly threefold increase of activity (p < 0.001) of the placental catalase (81.1 ± 20.6 U/mg of proteins in controls and 270.0 ± 69.9 U/mg in thrombophilic subjects), glutathione (GSH) peroxidase (C: 20.2 ± 10.1 U/mg; T: 60.0 ± 15.5 U/mg), and GSH reductase (C: 28.9 ± 5.6 U/mg; T: 72.7 ± 23.0 U/mg). The placenta...

  2. A Screen of FDA-Approved Drugs for Inhibitors of Zika Virus Infection. (United States)

    Barrows, Nicholas J; Campos, Rafael K; Powell, Steven T; Prasanth, K Reddisiva; Schott-Lerner, Geraldine; Soto-Acosta, Ruben; Galarza-Muñoz, Gaddiel; McGrath, Erica L; Urrabaz-Garza, Rheanna; Gao, Junling; Wu, Ping; Menon, Ramkumar; Saade, George; Fernandez-Salas, Ildefonso; Rossi, Shannan L; Vasilakis, Nikos; Routh, Andrew; Bradrick, Shelton S; Garcia-Blanco, Mariano A


    Currently there are no approved vaccines or specific therapies to prevent or treat Zika virus (ZIKV) infection. We interrogated a library of FDA-approved drugs for their ability to block infection of human HuH-7 cells by a newly isolated ZIKV strain (ZIKV MEX_I_7). More than 20 out of 774 tested compounds decreased ZIKV infection in our in vitro screening assay. Selected compounds were further validated for inhibition of ZIKV infection in human cervical, placental, and neural stem cell lines, as well as primary human amnion cells. Established anti-flaviviral drugs (e.g., bortezomib and mycophenolic acid) and others that had no previously known antiviral activity (e.g., daptomycin) were identified as inhibitors of ZIKV infection. Several drugs reduced ZIKV infection across multiple cell types. This study identifies drugs that could be tested in clinical studies of ZIKV infection and provides a resource of small molecules to study ZIKV pathogenesis. PMID:27476412

  3. Chorioamnionitis caused by Serratia marcescens in a non-immunocompromised host. (United States)

    Shimizu, S; Kojima, H; Yoshida, C; Suzukawa, K; Mukai, H Y; Hasegawa, Y; Hitomi, S; Nagasawa, T


    A 26 year old pregnant woman with antithrombin III deficiency developed recurrent septicaemia with Serratia marcescens. In spite of the administration of antibiotics, high grade fever persisted. She subsequently manifested lower abdominal pain, and spontaneous abortion occurred. After the abortion, she became completely afebrile. The amnion was turbid, and microscopic examination of the placenta showed haemorrhage and massive infiltration of neutrophils, suggestive of infectious chorioamnionitis. Pulsed field gel electrophoresis showed that isolates from the blood, urine, and vaginal discharge were genetically identical. Intravenous pyelography revealed that she had a bilateral completed double ureter. It was thought that a urinary tract anomaly caused infection with S marcescens, and the pathogen spread to the chorioamnion via the bloodstream. This is the first report of chorioamnionitis caused by S marcescens in a non-immunocompromised host. In addition, these findings indicate that the chorioamnion can serve as a site for persistent infection in normal pregnancies. PMID:14600137

  4. Placentation in dolphins from the Amazon River Basin: the Boto, Inia geoffrensis, and the Tucuxi, Sotalia fluviatilis

    Directory of Open Access Journals (Sweden)

    Bonatelli Marina


    Full Text Available Abstract A recent reassessment of the phylogenetic affinities of cetaceans makes it timely to compare their placentation with that of the artiodactyls. We studied the placentae of two sympatric species of dolphin from the Amazon River Basin, representing two distinct families. The umbilical cord branched to supply a bilobed allantoic sac. Small blood vessels and smooth muscle bundles were found within the stroma of the cord. Foci of squamous metaplasia occurred in the allanto-amnion and allantochorion. The interhemal membrane of the placenta was of the epitheliochorial type. Two different types of trophoblastic epithelium were seen. Most was of the simple columnar type and indented by fetal capillaries. However, there were also areolar regions with tall columnar trophoblast and these were more sparsely supplied with capillaries. The endometrium was well vascularised and richly supplied with actively secreting glands. These findings are consistent with the current view that Cetacea are nested within Artiodactyla as sister group to the hippopotamids.

  5. Quantitative mapping of intracellular cations in the human amniotic membrane (United States)

    Moretto, Ph.; Llabador, Y.; Simonoff, M.; Razafindrabe, L.; Bara, M.; Guiet-Bara, A.


    The effect of magnesium and taurine on the permeability of cell membranes to monovalent cations has been investigated using the Bordeaux nuclear microprobe. PIXE and RBS techniques have been used to provide quantitative measurements and ion distributions in the isolated amniotic membrane. This physiological model for cellular exchanges allowed us to reveal the distribution of most elements involved in cellular pathways and the modifications under different experimental conditions of incubation in physiological fluids. The PIXE microanalysis provided an original viewpoint on these mechanisms. Following this first study, the amnion compact lamina was found to play a role which was not, up to now, taken into account in the interpretation of electrophysiological experimentations. The release of some ionic species, such as K +, from the epithelial cells, during immersion in isotonic fluids, could have been hitherto underestimated.

  6. Hypoxic-ischemic encephalopathy with cystic brain stem necroses and thalamic calcifications in a preterm twin. (United States)

    Peters, B; Walka, M M; Friedmann, W; Stoltenburg-Didinger, G; Obladen, M


    A severe and rare ischemic brain lesion in a preterm twin boy is reported. The boy was born after two weeks of anhydramnios and amnionic infection at 24 weeks of gestation. Following a difficult Caesarean section and prolonged umbilical cord compression he developed prenatal acidosis with an umbilical cord pH of 6.96. At the age of 7 h, heart rate variability narrowed due to severely disturbed brain stem function and the patient developed clinical signs of hypoxic-ischemic encephalopathy. Sonography demonstrated extensive symmetrical brain stem and basal ganglia lesions. After a prolonged comatose and apneic state, death occurred at the age of 25 days. Autopsy confirmed columnar bilateral cavitation of basal ganglia, diencephalon, brain stem and spinal gray matter, as well as focal calcifications in the palladium, thalamus, and brain stem. The findings highly resemble those observed after experimental or clinical cardiac arrest.

  7. Mesenchymal stromal cells from the human placenta promote neovascularization in a mouse model in vivo. (United States)

    Kinzer, M; Hingerl, K; König, J; Reinisch, A; Strunk, D; Huppertz, B; Lang, I


    Cell transplantation is a promising strategy in regenerative medicine for revascularization of ischemic tissues. Based on our observation that placental mesenchymal stromal cells (PMSC) enhance endothelial cell viability in vitro via secretion of angiogenic factors, we asked whether PMSC support vascular growth in vivo. PMSC were isolated from amnion and placental endothelial cells (PLEC) from chorion and either separately or co-transplanted subcutaneously into immune-deficient mice. Co-transplantation resulted in a higher number of perfused human vessels (CD31+/vimentin+) containing mouse glycophorin A+ erythrocytes. Results indicate positive effects of PMSC on neovascularization in vivo, making them attractive candidates to create autologous PMSC/PLEC pairs for research and transplantation.

  8. Amniotic fluid and development of the immature intestine

    DEFF Research Database (Denmark)

    Østergaard, Mette Viberg

    in demand. Amniotic Fluid (AF) is the natural source of fetal EN throughout gestation in ammals. Fetal AF swallowing stimulates somatic and gastrointestinal growth during fetal development, and modulates the development of the intestinal mucosa. In addition, AF protects the fetus against infections......), and slow advancement of enteral feeds. Human breast milk is the gold standard of enteral nutrition (EN) of the newborn and protects against NEC in premature infants, but is often unavailable or limited after preterm birth. Innovative nutrition strategies and novel sources of the first EN are therefore...... and maintains homeostasis in the amnion sac through anti-inflammatory mechanisms. The growth promoting and protective effects of AF in utero are ascribed to bioactive proteins including growth factors, anti-inflammatory cytokines, and antimicrobial peptides, which may exert similar beneficial effects ex utero...

  9. Hypothetical role of prostaglandins in the onset of preterm labor after fetal surgery. (United States)

    Pomini, Francesco; Noia, Giuseppe; Mancuso, Salvatore


    Preterm labor is one of the most important factors limiting the advancement of fetal surgery programs. While prostaglandins (PGs) have long been indicated as the key factor in the initiation of labor in humans, there is significant evidence showing that the chorionic membrane acts as a powerful barrier between the decidua/myometrium and amniotic PGs during normal pregnancy. After either open or endoscopic fetal surgery the imperfect, non-hermetical closure of the chorion permits leakage of PGs from the amnionic sac, allowing them to reach the decidua and myometrium. The surgical wound in the chorionic barrier could be the major factor involved in preterm labor and delivery after human fetal surgery. PMID:17135752

  10. Oxytocinase-immunohistochemical demonstration in the immature and term human placenta. (United States)

    Small, C W; Watkins, W B


    Oxytocinase (cystine aminopeptidase) was purified from human retroplacental serum by a combination of fractional precipitation, hydroxylapatite chromatography and gel exlusion chromatography on Sephadex G-200. The purified enzyme possessed a specific activity of 980 mIU/mg using L-cystine-di-p-nitroanilide as substrate. This represented a 3200 fold concentration from the starting material in an overall yield of 12%. Antibodies against oxytocinase were raised in rabbits and the gamma-globulins fraction labelled with fluorescein isothiocyanate prior to its use in the immunofluorescence histochemical localization of the enzyme in human placental tissue. Oxytocinase was confined to the syncytiotrophoblastic cells of normal term, and immature placentas as well as in placentas from patients suffering from severe toxaemia. Specific immunofluorescence was also present in the outer margins of the chorion and to a lesser extent in the amnion.

  11. A Screen of FDA-Approved Drugs for Inhibitors of Zika Virus Infection. (United States)

    Barrows, Nicholas J; Campos, Rafael K; Powell, Steven T; Prasanth, K Reddisiva; Schott-Lerner, Geraldine; Soto-Acosta, Ruben; Galarza-Muñoz, Gaddiel; McGrath, Erica L; Urrabaz-Garza, Rheanna; Gao, Junling; Wu, Ping; Menon, Ramkumar; Saade, George; Fernandez-Salas, Ildefonso; Rossi, Shannan L; Vasilakis, Nikos; Routh, Andrew; Bradrick, Shelton S; Garcia-Blanco, Mariano A


    Currently there are no approved vaccines or specific therapies to prevent or treat Zika virus (ZIKV) infection. We interrogated a library of FDA-approved drugs for their ability to block infection of human HuH-7 cells by a newly isolated ZIKV strain (ZIKV MEX_I_7). More than 20 out of 774 tested compounds decreased ZIKV infection in our in vitro screening assay. Selected compounds were further validated for inhibition of ZIKV infection in human cervical, placental, and neural stem cell lines, as well as primary human amnion cells. Established anti-flaviviral drugs (e.g., bortezomib and mycophenolic acid) and others that had no previously known antiviral activity (e.g., daptomycin) were identified as inhibitors of ZIKV infection. Several drugs reduced ZIKV infection across multiple cell types. This study identifies drugs that could be tested in clinical studies of ZIKV infection and provides a resource of small molecules to study ZIKV pathogenesis.

  12. Velamentous Cord Insertion in a Singleton Pregnancy: An Obscure Cause of Emergency Cesarean—A Case Report

    Directory of Open Access Journals (Sweden)

    Juliana Rocha


    Full Text Available Approximately 2% of low-risk pregnant women still require an emergency Cesarean section after the onset of labor. Because it is likely that half of these cases are associated with placental and umbilical cord abnormalities, it is thought that prenatal detection of such abnormalities would reduce the number of emergency cesarean sections in low-risk women. Velamentous cord insertion is an abnormal cord insertion in which the umbilical vessels diverge as they traverse between the amnion and chorion before reaching the placenta. With a reported incidence of 1% in singleton pregnancies, it has been associated with several obstetric complications. This condition has been diagnosed by ultrasonography with a sensitivity of 67% and specificity of 100% in the second trimester. The present case highlights the importance of the systematic assessment of the placental cord insertion site at routine obstetric ultrasound and the potential of identifying pregnancies with velamentous insertion and, therefore, those at risk for obstetric complications.

  13. Amniotic band syndrome (ABS): can something be done during pregnancy in African poor countries? Three cases and review of the literature. (United States)

    Mian, D B; Nguessan, K L P; Aissi, G; Boni, S


    Amniotic band syndrome (ABS) is a fetal congenital malformation, affecting mainly the limbs, but also the craniofacial area and internal organs. Two mains pathogenic mechanisms are proposed in its genesis. Firstly the early amnion rupture (exogenous theory) leading to fibrous bands, which wrap up the fetal body; secondly, the endogenous theory privileges vascular origin, mesoblastic strings not being a causal agent. The authors believe that the second theory explain the occurrence of ABS. The outcome of the disease during pregnancy depends on the gravity of the malformations. Interruption of the pregnancy is usually proposed when diagnosis of severe craniofacial and visceral abnormalities is confirmed. Whereas minor limb defects can be repaired with postnatal surgery. In case of an isolated amniotic band with a constricted limb, in utero lysis of the band can be considered to avoid a natural amputation. In an African country, such treatment is not possible as far as the antenatal diagnosis. PMID:24779260

  14. Therapeutic outcomes of transplantation of amniotic fluid-derived stem cells in experimental ischemic stroke

    Directory of Open Access Journals (Sweden)

    Naoki eTajiri


    Full Text Available Accumulating preclinical evidence suggests the use of amnion as a source of stem cells for investigations of basic science concepts related to developmental cell biology, but also for stem cells’ therapeutic applications in treating human disorders. We previously reported isolation of viable rat amniotic fluid-derived stem (AFS cells. Subsequently, we recently reported the therapeutic benefits of intravenous transplantation of AFS cells in a rodent model of ischemic stroke. Parallel lines of investiagtions have provided safety and efficacy of stem cell therapy for treating stroke and other neurological disorders. This review article highlights characterization of AFS cells’ phenotype and their transplant-mediated functional effects, the need for investigations of mechanisms underlying AFS cells’ therapeutic benefits and discusses lab-to-clinic translational gating items in an effort to optimize the clinical application of cell transplantation for stroke.

  15. Human amniotic epithelial cells express specific markers of nerve cells and migrate along the nerve fibers in the corpus callosum

    Institute of Scientific and Technical Information of China (English)

    Zhiyuan Wu; Guozhen Hui; Yi Lu; Tianjin Liu; Qin Huang; Lihe Guo


    Human amniotic epithelial cells were isolated from a piece of fresh amnion. Using immunocytochemical methods, we investigated the expression of neuronal phenotypes (microtubule-associated protein-2, glial fibrillary acidic protein and nestin) in human amniotic epithelial cells. The conditioned medium of human amniotic epithelial cells promoted the growth and proliferation of rat glial cells cultured in vitro, and this effect was dose-dependent. Human amniotic epithelial cells were further transplanted into the corpus striatum of healthy adult rats and the grafted cells could integrate with the host and migrate 1-2 mm along the nerve fibers in corpus callosum. Our experimental findings indicate that human amniotic epithelial cells may be a new kind of seed cells for use in neurograft.

  16. Tissue banking and clinical research on radiation and ethylene oxide sterilization of tissue grafts

    International Nuclear Information System (INIS)

    The research works carried out in Rangoon, Burma under the Agency supported project RC4420/RB have dealt with an elucidation of the radiation interaction(s) with the species of biomolecules such as proteins, lipids, collagens, connective tissues present in the cleaned and freeze-dried non-viable tissue grafts. Radiation as a cool process furthermore effectively helps to destroy the microbial bioburden as the undesirable contaminants which may associate the tissue grafts. Radiation also concomitantly helps to suppress the tissue-specific immunogenicity. All these attributes of radiation induced effects have proved successful towards the development of a sterilization process. A series of non-viable tissue grafts, such as bone, nerve, fascia, dura, cartilage, chorion-amnion (as dressings in burn wounds) and tympanic membrane have been successfully attempted in Burma and many more possibilities seem to still remain unexplored. Radiation sterilization modality has proved as a blessing for the promotion of clinical surgical applications of tissue allografts in the corrective/reconstructive surgery on the disability cases due to diseases which accompany tissue losses. The investigator in Burma has reported on the case histories where freeze dried radiation sterilized tissue allografts have been successfully used in the osteogenic inductions (bone grafts); midear tympanoplasty; partial recovery of nerve sensation throught nerve allografts; rapid healing of high degree burn wounds through the use of amnion dressings. Besides, there have been a widespread surgical use of radiation sterilized dura and fascia as allografts. A national tissue banking facility has been established in Burma surrounding the processing and clinical utilization of tissue allografts which has involved over ten hospital centres throughout the country. Radiation induced effects on the biomolecules of clinical significance in the tissue grafts have been researched to help gain insight into a better

  17. Preterm labor and preterm premature rupture of membranes have a different pattern in the involved compartments of acute histologoic chorioamnionitis and/or funisitis: Patho-physiologic implication related to different clinical manifestations. (United States)

    Park, Chan-Wook; Park, Joong Shin; Moon, Kyung Chul; Jun, Jong Kwan; Yoon, Bo Hyun


    It is unknown whether histo-topographic findings about the involved compartments (i.e., choriodecidua, amnion, chorionic-plate) of acute-histologic chorioamnionitis (acute-HCA) and/or funisitis according to the presence or absence of intra-amniotic inflammation (IAI) and/or fetal inflammatory response syndrome (FIRS) are different between preterm labor and intact membranes (PTL) and preterm premature rupture of membranes (preterm-PROM). The involved compartments of acute-HCA and/or funisitis were examined in 161 singleton preterm-births (preterm-PROM (n = 73). The study-population was divided into IAI(-)/FIRS(-), IAI(+)/FIRS(-), and IAI(+)/FIRS(+) groups according to the presence or absence of IAI (amniotic-fluid MMP-8 ≥ 23 ng/ml) and/or FIRS (umbilical-cord plasma CRP ≥ 200 ng/ml). Histological inflammation was not detected in any-compartment except choriodecidua in IAI(-)/FIRS(-) group with PTL while inflammation appeared in all-compartment0s (choriodeciduitis-46.2 %; amnionitis-23.1 %; funisitis-30.8 %; chorionic-plate inflammation-7.7 %) in IAI(-)/FIRS(-) group with preterm-PROM. IAI(+)/FIRS(-) group had a significantly higher frequency of inflammation in each-compartment than IAI(-)/FIRS(-) group in PTL (each-for P preterm-PROM (each-for P > 0.1). However, IAI(+)/FIRS(+) group had a significantly higher rate of inflammation in each compartment than IAI(+)/FIRS(-) group in both PTL and preterm-PROM (each-for P preterm-PROM had a different pattern in the involved compartments of acute-HCA and/or funisitis in the IAI(-)/FIRS(--) group and in the change of involved compartments from IAI(-)/FIRS(-) to IAI(+)/FIRS(-).

  18. Bronchial stump closure with amniotic membrane in animal model

    Directory of Open Access Journals (Sweden)

    Gholamreza Mohajeri


    Full Text Available Background: Coverage of the bronchial stumps (BSs with adjacent tissues can improve healing and reduce bronchial complications in complex thoracic surgery. There is no evidence for the application of human amnion allograft for prevention of air leak from the BS. The comparison of the amniotic membrane (AM and pleural patch for BS healing after lobectomy in dogs was our aim in this study. Materials and Methods: A total of eight males and females 12-24-month-old dogs between 17 and 22 kg body-weight were used in this study in 2010, Isfahan University of Medical Sciences. Animals were separated into two groups: group A (n = 4; amniotic membrane and group P (n = 4; pleural patch according to the BS closure technique performed. After lobectomy of the right middle lobe, the BS was closed, while a small bronchopleural fistula (BPF was created by inserting a catheter via edges of closed stump. Then, it was covered with a piece of AM3 × 3 cm in group A and with a pedicle graft of pleura in group P. Rethoracotomy was performed after 15 days of observation, and the BS was removed for histological examination. Histological healing was classified as complete or incomplete healing. Neoangiogenesis was measured by Von Willebrand expression using immunohistochemistry (IHC. Data were analyzed by SPSS version 15 using Fisher′s exact test, Mann-Whitney test, and T tests. Results: BPF complications were not seen during observation period. There was no significant difference in histological healing between two groups. Similarly, no significant difference was observed between the groups in terms of neoangiogenesis based on IHC examination (P value = 0.69. Conclusion: Human amnion allograft could be as effective as pleural patch for BS wrapping following pulmonary resections.

  19. The impact of the International Atomic Energy Agency (IAEA) program on radiation and tissue banking in Peru. (United States)

    Gamero, Emma Castro; Morales Pedraza, Jorge


    The tissue bank "Rosa Guerzoni Chambergo" (RGCTB) located at the Child's Health Institute was inaugurated in 1996, with the financial and technical support of the IAEA program on radiation and tissue banking. Since 1998, the biological bandage of fresh and lyophilised pigskin, amnion and bone tissue is processed routinely in this bank. In all cases, the tissue is sterilised with the use of Cobalt-60 radiation, process carried out at the Laboratories of Irradiation of the Peruvian Institute of Nuclear Energy (IPEN). The tissue bank in the Child's Health Institute helped to save lives in an accident occurred in Lima, when a New Year's fireworks celebration ran out of control in January 2002. Nearly 300 people died in the tragic blaze and hundreds more were seriously burned and injured. Eight Lima hospitals and clinics suddenly were faced with saving the lives of severely burned men, women and children. Fortunately, authorities were ready to respond to the emergency. More than 1,600 dressings were sterilised and supplied to Lima surgeons. The efforts helped save the lives of patients who otherwise might not have survived the Lima fire. Between 1998 and September 2007, 35,012 tissue grafts were produced and irradiated. Radiation sterilised tissues are used by 20 national medical institutions as well as 17 private health institutions. The tissue bank established in Peru with the support of the IAEA is now producing the following tissues: pigskin dressings, fresh and freeze-dried; bone allografts, chips, wedges and powdered, and amnion dressings air-dried. It is also now leading the elaboration of national standards, assignment being entrusted by ONDT (Organización Nacional de Donación y Transplantes; National Organisation on Donation and Transplant). This among other will permit the accreditation of the tissue bank. In this task is also participating IPEN. PMID:18612849

  20. Innate immune defences in the human uterus during pregnancy. (United States)

    King, A E; Kelly, R W; Sallenave, J-M; Bocking, A D; Challis, J R G


    The prevention of uterine infection is critical to appropriate fetal development and term delivery. The innate immune system is one component of the uterine environment and has a role in prevention of uterine infection. Natural antimicrobials are innate immune molecules with anti-bacterial, anti-viral and anti-fungal activity. We discuss two groups of natural antimicrobials in relation to pregnancy: (i) the defensins; and (ii) the whey acidic protein motif containing proteins, secretory leukocyte protease inhibitor (SLPI) and elafin. Human beta-defensins (HBD) 1-3 are expressed by placental and chorion trophoblast, amnion epithelium and decidua in term and preterm pregnancy. Elafin shows a similar pattern of localisation while SLPI is produced only by amnion epithelium and decidua. Evidence suggests that there is aberrant production of some natural antimicrobials in pathologic conditions of pregnancy. In preterm premature rupture of membranes (PPROM) levels of SLPI and elafin are reduced in amniotic fluid and fetal membranes, respectively. Elafin and HBD3 increase in chorioamnionitis and levels of the alpha-defensins, HNP1-3, increase in maternal plasma and amniotic fluid in women affected by microbial invasion of the uterus. In vitro culture studies have suggested a mechanism for increased production of natural antimicrobials in chorioamnionitis. Elafin, SLPI, HBD2 and 3 are all upregulated by inflammatory molecules in cells derived from gestational tissues. In summary, production of natural antimicrobials at key sites within the pregnant uterus suggests an important role in prevention of uterine infection during pregnancy and labour. Aberrant production of these molecules in PPROM and chorioamnionitis suggests that they also have a role in pathologic conditions. In particular, upregulation of these molecules by inflammatory molecules present in chorioamnionitis will ensure a robust response to infection. PMID:17664005

  1. Nuclear factor-kappa B localization and function within intrauterine tissues from term and preterm labor and cultured fetal membranes

    Directory of Open Access Journals (Sweden)

    Kusanovic Juan P


    Full Text Available Abstract Background The objective of this study was to quantify the nuclear localization and DNA binding activity of p65, the major transactivating nuclear factor-kappa B (NF-kappaB subunit, in full-thickness fetal membranes (FM and myometrium in the absence or presence of term or preterm labor. Methods Paired full-thickness FM and myometrial samples were collected from women in the following cohorts: preterm no labor (PNL, N = 22, spontaneous preterm labor (PTL, N = 21, term no labor (TNL, N = 23, and spontaneous term labor (STL, N = 21. NF-kappaB p65 localization was assessed by immunohistochemistry, and DNA binding activity was evaluated using an enzyme-linked immunosorbent assay (ELISA-based method. Results Nuclear p65 labeling was rare in amnion and chorion, irrespective of clinical context. In decidua, nuclear p65 labeling was greater in the STL group relative to the TNL cohort, but there were no differences among the TNL, PTL, and PNL cohorts. In myometrium, diffuse p65 nuclear labeling was significantly associated with both term and preterm labor. There were no significant differences in ELISA-based p65 binding activity in amnion, choriodecidual, and myometrial specimens in the absence or presence of term labor. However, parallel experiments using cultured term fetal membranes demonstrated high levels of p65-like binding even the absence of cytokine stimulation, suggesting that this assay may be of limited value when applied to tissue specimens. Conclusions These results suggest that the decidua is an important site of NF-kappaB regulation in fetal membranes, and that mechanisms other than cytoplasmic sequestration may limit NF-kappaB activation prior to term.

  2. The tissue bank at the national nuclear research institute in Mexico. (United States)

    Esther Martínez-Pardo, María; Lourdes Reyes-Frías, Ma


    The Instituto Nacional de Investigaciones Nucleares (ININ, The National Nuclear Research Institute) received during 1997-1998 strong support of the International Atomic Energy Agency (IAEA), to establish the first and only one tissue bank (BTR ININ tissue bank) in Mexico that uses ionising radiation as sterilising agent. In that time, the BTR staff was trained in different tissue banks in several countries. Basic equipment for tissue processing donated by the IAEA was received in 1998. In July, 1999 the Mexican Health Secretariat gave the Sanitary License No. 1062000001 to the BTR to operate as an official organ and tissue bank. In August, 2001 the ININ and the Hospital Materno Infantil (HMI-ISSEMYM) signed an agreement to collaborate in amnion processing. The hospital is responsible for donor selection, serology tests, tissue procurement and washing, since this hospital is the BTR amnion supplier. The tissues are collected by ININ weekly with complete documentation. The BTR is responsible for processing: cleaning, air drying, packaging, labelling, microbiological control and sterilisation by gamma irradiation. The sterilised tissue is kept under quarantine for 6 months to obtain the results of the donor second serology test. From March to June, 2002 the BTR has processed 347.86 units (50 cm(2) each), is say, 17,393 cm(2). In addition, the pig skin xenograft process has been implemented and a protocol for clinical applications of it is running at the Hospital Central Sur de Alta Especialidad (PEMEX). Also the ININ tissue bank present status and perspectives are described. PMID:15256855

  3. About the tactics of treatment and visual rehabilitation in case of an acute severe alkaline burn injury of the eye. A case report

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    V.F. Chernysh


    Full Text Available ABSTRACT Purpose. To acquaint ophthalmologists with tactics of treatment of a victim with an acute severe chemical burn injury of the eye i n one of follow-up cases fr om practice. Material and methods. The female patient of 31 years old had a severe alkaline burn injury with total limbal stem cell deficiency in the right eye. до (до After an emergency aid an ischemia of a limbus zone, a total corneal erosion and its moderate edema, a deepithelization of conjunctiva up to the fornices were observed. Visual acuity was 0.3 (before the burn it was 1.0. At 8 days after the burn the covering of ocular surface structures without epithelium was performed using a fresh amnion transplant which edges were sutured closely to the edges of intact bulbar conjunctiva. The total epithelization of the ocular surface (including the cornea was noted at 39 days after the operation. Visual acuity was 0.2. One year later the visual acuity was 0.07 and the total fibrovascular pannus with the effects of corneal epitheliopathy was detected. The сonjunctival limbal autograft was carried out. Six months later the visual acuity w as 0.9. Conclusion. During the early periods of ocular burn disease with the total limbal stem cell deficiency, the covering of the ocular surface using amnion in a variant of conjunctival plasty reduced the healing time, stabilized the condition of the corneal stroma after the burn, prevented the corneal opacity and an inevitable vascular in-growth creating favorable conditions for the limbal stem cell transplantation.

  4. 红花注射液保存人胎羊膜对大鼠皮肤切创愈合中MMP-2,MMP-9影响的研究*%Expressions of MMP-2,MMP-9 in Rat Skin Incised Wound Affected by Human Fetal Amniotic Preserved by Safflower Injection

    Institute of Scientific and Technical Information of China (English)

    王君; 罗秋燕; 边文玲; 申素芳


      目的:研究经红花注射液保存后的人胎羊膜对大鼠皮肤切创愈合过程中基质金属蛋白酶(Matrix metalloproteinase-s, MMPS)-2,9表达的影响,探讨其促进切口愈合的机制,为其在临床的应用提供理论依据。方法:建立大鼠皮肤切创模型,观察切口愈合时间、感染率;在各时相点处死取材,应用免疫组织化学技术,通过计算机图像分析系统检测人胎羊膜对不同损伤时间大鼠皮肤组织中MMP-2, MMP-9的表达。结果:创后12 h、1 d、2 d、3 d,红花+羊膜组创缘周围MMP-2,MMP-9表达,较其他三组明显增强(P<0.05);红花+羊膜组感染率(5%)较其他三组明显减少(P<0.05)。结论:经红花注射液保存后的人胎羊膜在大鼠皮肤切创愈合过程中,可促进创缘周围MMP-2,MMP-9的表达;将其贴附创面能明显减少创口的感染率、促进创口的愈合。%  Objective:To observe the infection rate and MMP-2,MMP-9 expression in rat skin incised wound healing process with human fetal amnion preserved by the safflower injection. Explore the mechanism of skin healing process and reduce scar formation. Method:Rats were executed at different time after models of rats’skin incision were set,and then used immunohistochemical and auto image analysis system to detect the infection rate and MMP-2,MMP-9 expression in group of human amniotic membrane saved with safflower injection at different traumatic time intervals. Result:The expressions of MMP-2,MMP-9 was much higher in group of human amniotic membrane saved with safflower injection than others(P<0.05);the infection rate of the human amnion saved with safflower injection group(5%)was less than that of the other groups(P<0.05). Conclusion:The human amnion preserved by safflower injection could not only enhance the expression of MMP-2,MMP-9 around the area of injury,but also could reduce the infection rate of incised wound skin of rat

  5. Clinical observatiom of corneal limbal stem cell auto-grafting,the transplantation of anmion and the pure removal of pterygium to treat the recurrent pterygium%角膜缘干细胞移植与羊膜移植及单纯胬肉切除治疗复发性翼状胬肉的疗效比较

    Institute of Scientific and Technical Information of China (English)

    唐茂聪; 唐铮; 沈亚玲


    Objective The effect analyses by comparing and observing corneal limbal stem cell auto-grafting,the transplantation of flesh amnlon and the pure removal of pterygium to treat the recurrent pterygium.Method 166 cases of recurrent Pterygium( 176 eyes)is divides into 3 groups(named A,B,and C) randomly.Group A (with 67 cases, 72 eyes)is operated by removing the recurrent pterygium and corneal limbal stem cell auto-grafting.Group B (with 58 cases,59 eyes)is operated by removing the recurrent pterygium and the transplantation of flesh amnion.Group C (with 43 eases,45 eyes)is only operated by removing the pterygium.After operation,visits lasting 6-60 months are followed and comparations about the patients' symptoms,the wound matches and the pterygium relapses among the 3 groups are carried out.Result In group A,28 eyes relapsed from the whole 72 eyes,and the rate of relapsing is 39%.In group B,28 eyes relapsed from the whole 59 eyes,and the rate of relapsing is 47.5%.In group C,39 eyes relapsed from the whole 45 eyes,with the rate of relapsing 86.6%.Conclusion Compared with the effect of the pure removal pterygium to treat recurrent pterygium, there exist lower rates of relapsing in corneal limbai stem cell autografting and the transplantation of fresh amnion.Corneal limbal stem cell auto-grafting is more effective than the transplantation of flesh amnion in lowering the rate of relapsing recurrent pterygium.In addition, corneal iimbal stem cells are easily to be found and economical,and it is the effective option in treating recurrent pterygium.%目的 比较自体角膜缘干细胞移植与新鲜羊膜移植及单纯胬肉切除治疗复发性翼状胬肉的疗效.方法 对复发性翼状胬肉166例(176只眼),随机分为A、B、C 3组,A组67例,72只眼,行复发性翼状胬肉切除联合自体角膜缘干细胞移植术.B组58例,59只眼,行复发性翼状胬肉切除联合新鲜羊膜移植术.C组43例,45只眼,行单纯胬肉切除.手术后随访6~60

  6. Tissue-specific human beta-defensins (HBD-1, HBD-2 and HBD-3 secretion profile from human amniochorionic membranes stimulated with Candida albicans in a two-compartment tissue culture system

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    Zaga-Clavellina Veronica


    Full Text Available Abstract Background During intrauterine infection, amniochorionic membranes represent a mechanical and immunological barrier against dissemination of infection. Human beta defensins (HBD-1, HBD-2, and HBD-3 are key elements of innate immunity that represent the first line of defense against different pathogen microorganisms associated with preterm labor. The aim of this work was to characterize the individual contribution of the amnion (AMN and choriodecidua (CHD regions to the secretion of HBD-1, HBD-2 and HBD-3, after stimulation with Candida albicans. Methods Full-thickness human amniochorionic membranes were obtained after delivery by elective cesarean section from women at 37-40 wk of gestation with no evidence of active labor. The membranes were cultured in a two-compartment experimental model in which the upper compartment is delimited by the amnion and the lower chamber by the choriodecidual membrane. One million of Candida albicans were added to either the AMN or the CHD face or to both and compartmentalized secretion profiles of HBD-1, HBD-2, and HBD-3 were quantified by ELISA. Tissue immunolocalization was performed to detect the presence of HBD-1, -2, -3 in tissue sections stimulated with Candida albicans. Results HBD-1 secretion level by the CHD compartment increased 2.6 times (27.30 [20.9-38.25] pg/micrograms protein when the stimulus with Candida albicans was applied only on this side of the membrane and 2.4 times (26.55 [19.4-42.5] pg/micrograms protein when applied to both compartments simultaneously. HBD-1 in the amniotic compartment remained without significant changes. HBD-2 secretion level increased significantly in the CHD when the stimulus was applied only to this region (2.49 [1.49-2.95] pg/micrograms protein and simultaneously to both compartments (2.14 [1.67- 2.91] pg/micrograms protein. When the stimulus was done in the amniotic compartment HBD-2 remained without significant changes in both compartments. HBD-3 remained

  7. Elaboration of amniotic membrane dressing dried by air and irradiated - Peruvian experience

    International Nuclear Information System (INIS)

    The purpose of this work is to prepare dressings from the amniotic membrane to be used in cases of skin damage principally due to superficial and intermediate second-degree burns. The amnion is a transparent membrane that lines the chorion. It is resistant and rich in collagen. Due to these characteristics it can be well used as biological dressing as it diminishes the loss of fluids, electrolytes and proteins, it also protects the growing epithelium and adheres well to the surface of the wound, improves mobility of the patient, diminishing pain and stimulating neovascularization. The ISN-IPEN Tissue Bank promoted by IAEA has processed amniotic membrane since July 1997. Initially dressings were prepared using antibiotics, after IAEA training at the MINT of Malaysia, it is processed dried by air, lyophylized and in both presentations, sterilized by gamma-rays. Amniotic membranes are procured from Lima Maternity. Tissues must comply with VDRL, HIV, Hepatitis B and C exclusion tests. The process is held in a laminar flow hood and amnion already separated from the chorion is washed with sterile distilled water, a solution of 0.05% sodium hypochlorite, and normal saline. Then it is cut into appropriate sizes and double packed in PE films. The dressings are then carried to the Peruvian Institute of Nuclear Energy for irradiation, depending on the number of samples either irradiated with gamma-rays at the Gammacell 220 or at the Irradiation Facility located in Santa Anita. The delivered dose is 25 kGy. The product is only released if it complies with the end product quality controls. Meanwhile, microbiological tests are carried out during all the processing stages, in order to monitor the microbial load during production. In conclusion we can state that dressings prepared as above mentioned have the following advantages: not complicated preparation; reliable and safe for clinical use; diminish infection rates and days spent in the hospital; easy to storage; and can be

  8. Isolation and biological characteristics of three kinds of human placenta-derived mesenchymal stem cells%三种胎盘间充质干细胞的分离培养及生物学特性对比研究

    Institute of Scientific and Technical Information of China (English)

    刘丽; 黎渊明; 刘琴; 张亚光; 毛圆圆; 宋云庆


    目的 分离培养人胎盘羊膜、绒毛膜和蜕膜三种间充质干细胞(mesenchymal stem cells,MSCs),进行生物学特性对比研究.方法 酶消化法分离胎盘羊膜、绒毛膜和蜕膜内干细胞,体外培养传代,MTT法测定细胞生长曲线,流式细胞术检测细胞表面标志,茜素红和油红O检测成骨和成脂能力.结果 表面标记鉴定分离的胎盘细胞为间充质干细胞,细胞具增殖和分化能力.三种细胞在传代过程中细胞形态、表面标志表达、分化能力维持稳定,但增殖能力存在差异.结论 成功分离获得三种胎盘MSCs,可以作为临床前研究用MSCs来源.%Objective To isolate mesenchymal stem cells from human amnions, chorions and decidua basalis, and compare their biological characteristics. Methods Stem cells were obtained by enzymatic digestion method from human amnions, chorions and decidua basalis and cultured in vitro. Cell growth curves were made by MIT method. Cell surface markers were assayed by flow cytometry. Osteogenic and adipogenic differentiation capacity was determined by alizarin red and alizarin red staining. Results Three kinds of placenta-derived mesenchymal stem cells were confirmed by cell surface markers assay. And these cells possess proliferation and differentiation ability. Cell morphology, surface markers and differentiation ability maintained stability during passage. However difference was detected in growth ability. Conclusion Three kinds of human placenta-derived stem cells were isolated and cultured successfully, and they could be used as seed cells in later preclinical study.

  9. 组织工程角膜上皮移植抑制角膜碱烧伤新生血管形成的临床观察%Clinical evaluation of tissue-engineered corneal epithelium transplantation in suppressing corneal neovascularization caused by alkali burns

    Institute of Scientific and Technical Information of China (English)

    郭青; 皮裕琍; 董莹; 尹澜


    Objective:To evaluate the clinical effects of tissue-engineered epithelium transplantation on suppressing corneal neovascularization as a sequela of alkali burn. Methods: Among 17 cases (24 eyes) of limbal stem cell deficiency caused by severe alkali burns from 2005 to 2010,9 cases (11 eyes) underwent tissue-engineered epithelium transplantation, and 8 cases (13 eyes) received amnionic membrane transplantation. The status of ocular surface, stromal infiltration and vascularization was observed with slit-lamp microscope in all patients. A neovascularization scoring system was used to evaluate the conditions of each eye before and after surgery. Corneal neovascularization scores in both two groups were evaluated. Results: Corneal neovascularization scores were decreased significantly after tissue-engineered epithelium transplantation or amniotic membrane transplantation ( P 0. 05) , indicating that neovascularization became stable 3 weeks after tissue-engineered epithelium transplantation or amniotic membrane transplantation. Conclusions: Tissue-engineered epithelium transplantation was more effective than amnion membrane transplantation in suppressing the growth of corneal neovascularization as a sequela of alkali burn.%目的:观察组织工程角膜上皮移植对角膜碱烧伤新生血管的抑制作用.方法:我院2005-2010年期间收治严重角膜碱烧伤(角膜缘干细胞缺陷)17例24眼,其中组织工程角膜上皮移植9例11眼,羊膜移植8例13眼.所有患者在手术前及手术后均行裂隙灯检查,观察角膜上皮、组织浸润、新生血管情况,对两组患者角膜新生血管情况进行评分和比较.结果:术后两组角膜新生血管评分均较手术前明显减少(P0.05),表明两组角膜新生血管在移植3周后基本保持稳定.结论:组织工程角膜上皮移植对严重的角膜碱烧伤新生血管的抑制作用明显好于羊膜移植.

  10. The amniotic membrane as a source of stem cells. (United States)

    Insausti, Carmen L; Blanquer, Miguel; Bleda, Patricia; Iniesta, Paqui; Majado, María J; Castellanos, Gregorio; Moraleda, José M


    Cellular therapy has emerged as a new potential tool for curing a wide range of degenerative diseases and tissue necrosis. Embryonic stem cells possess potential for differentiation into a wide range of cell lineages, but the ethical issues associated with establishment of this human cell line have to be resolved prior to any use. The bone marrow (BM) is the usual source of adult stem cells for hematopoietic stem cell transplants and cellular therapy, but the BM harvest is a surgical procedure that requires general anesthesia or sedation, and there seems to be a reduction of the proliferative potential and differentiation capacity of the marrow mesenchymal stem cells in older donors. For these reasons there is an increasing interest in other sources of stem cells from adult and fetal tissues. The amniotic membrane (AM) or amnion is a tissue of particular interest because its cells possess characteristics of stem cells with multipotent differentiation ability, and because of low immunogenicity and easy procurement from the placenta, which is a discarded tissue after parturition, thus avoiding the current controversies associated with the use of human embryonic stem cells. Therefore, amniotic membrane has been proposed as a good candidate to be used in cellular therapy and regenerative medicine. PMID:19924645

  11. Release of LHRH-activity from human fetal membranes upon exposure to PGE/sub 2/, oxytocin and isoproterenol

    Energy Technology Data Exchange (ETDEWEB)

    Poisner, A.M.; Poisner, R.; Becca, C.R.; Conn, P.M.


    The authors have previously reported that superfused chorion laeve (fetal membranes) release LHRH-like immunoreactivity upon exposure to angiotensin II. They have now studied the effects of other agonists on the release of LHRH-activity and something of its chemical nature. Fetal membranes were obtained from placentas delivered by cesarean section, the amnion stripped from the chorion, and the chorion superfused in an Amicon thin-channel device with the maternal surface facing up. The whole device was submerged in a 37 C water bath and perfused with a modified Locke's solution at 0.4 - 1.0 ml/min. LHRH-activity was measured by radioimmunoassay using three different antisera against LHRH. The release of LHRH-activity was stimulated by 6-10 min exposure to PGE/sub 2/, oxytocin, and isoproterenol. Extracts of chorion were studied using gel filtration on Sephacryl S-200 and ultrafiltration with Amicon PM-10 filters. The bulk of the LHRH-activity appeared as a higher molecular weight form (about 70,000 daltons). Since oxytocin has been reported to release PGE/sub 2/ from chorion, it may release LHRH-activity by virtue of liberating endogenous PGE/sub 2/. The chemical nature of the LHRH-activity is presently under investigation.

  12. Efficacy of Tectonic Corneal Patch Graft for Progressive Peripheral Corneal Thinning

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    Cafer Tanrıverdio


    Full Text Available Objectives: To report the results of tectonic corneal patch graft (TCPG in patients with progressive peripheral corneal thinning (PCT. Materials and Methods: In this study, we included 8 patients who underwent TCPG for PCT or perforated corneal ulceration at Ankara Training and Research Hospital. Results: We performed TCPG in 7 patients for PCT and in 1 patient for perforated corneal ulceration. Mean age was 57.2±16.7 (38- 82 years. Postoperative follow-up time ranged from 6 to 24 months (mean 13.9±6.7. Possible etiologies leading to progressive PCT were trachoma, infectious corneal ulcer, and rheumatoid arthritis-severe dry eye in 2 patients each. Other 2 patients had a progressive PCT following ocular surgery. One of the patients with infectious corneal ulcer also had a trauma caused by a scissor. Amnion membrane transplantation was performed in 3 patients prior to TCPG. While the anatomic success was achieved in all 8 patients, best-corrected visual acuity (BCVA was 0.1 or better in 4 patients (50%. Postoperative BCVA was better than preoperative BCVA in 6 patients (75%. Local peripheral anterior synechiae developed in two eyes. Conclusion: TCPG is a useful therapeutic option in selected cases of corneal thinning and perforations because it effectively restores the integrity of the globe and allows acceptable visual results. (Turk J Ophthalmol 2014; 44: 440-4

  13. [Biomechanical characteristics of human fetal membranes. Preterm fetal membranes are stronger than term fetal membranes]. (United States)

    Rangaswamy, N; Abdelrahim, A; Moore, R M; Uyen, L; Mercer, B M; Mansour, J M; Kumar, D; Sawady, J; Moore, J J


    The purpose of this study was to determine the biomechanical characteristics of human fetal membranes (FM) throughout gestation. Biomechanical properties were determined for 115 FM of 23-41 weeks gestation using our previously described methodology. The areas of membrane immediately adjacent to the strongest and weakest tested spots were sampled for histomorphometric analysis. Clinical data on the patients whose FM were examined were also collected. FM less than 28 weeks gestation were associated with higher incidence of abruption and chorioamnionitis. Topographically FM at all gestations had heterogeneous biomechanical characteristics over their surfaces with distinct weak areas. The most premature membranes were the strongest. FM strength represented by rupture force and work to rupture decreased with increasing gestation in both weak and strong regions of FM. This decrease in FM strength was most dramatic at more than 38 weeks gestation. The FM component amnion-chorion sublayers were thinner in the weak areas compared to strong areas. Compared to term FM, preterm FM are stronger but have similar heterogeneous weak and strong areas. Following a gradual increase in FM weakness with increasing gestation, there is a major drop-off at term 38 weeks gestation. The FM weak areas are thinner than the stronger areas. Whether the difference in thickness is enough to account for the strength differences is unknown.

  14. Regulation of amniotic fluid volume: mathematical model based on intramembranous transport mechanisms. (United States)

    Brace, Robert A; Anderson, Debra F; Cheung, Cecilia Y


    Experimentation in late-gestation fetal sheep has suggested that regulation of amniotic fluid (AF) volume occurs primarily by modulating the rate of intramembranous transport of water and solutes across the amnion into underlying fetal blood vessels. In order to gain insight into intramembranous transport mechanisms, we developed a computer model that allows simulation of experimentally measured changes in AF volume and composition over time. The model included fetal urine excretion and lung liquid secretion as inflows into the amniotic compartment plus fetal swallowing and intramembranous absorption as outflows. By using experimental flows and solute concentrations for urine, lung liquid, and swallowed fluid in combination with the passive and active transport mechanisms of the intramembranous pathway, we simulated AF responses to basal conditions, intra-amniotic fluid infusions, fetal intravascular infusions, urine replacement, and tracheoesophageal occlusion. The experimental data are consistent with four intramembranous transport mechanisms acting in concert: 1) an active unidirectional bulk transport of AF with all dissolved solutes out of AF into fetal blood presumably by vesicles; 2) passive bidirectional diffusion of solutes, such as sodium and chloride, between fetal blood and AF; 3) passive bidirectional water movement between AF and fetal blood; and 4) unidirectional transport of lactate into the AF. Further, only unidirectional bulk transport is dynamically regulated. The simulations also identified areas for future study: 1) identifying intramembranous stimulators and inhibitors, 2) determining the semipermeability characteristics of the intramembranous pathway, and 3) characterizing the vesicles that are the primary mediators of intramembranous transport.

  15. Antioxidative Defense Enzymes in Placenta Protect Placenta and Fetus in Inherited Thrombophilia from Hydrogen Peroxide

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    Jelena Bogdanovic Pristov


    Full Text Available Our aim was to investigate the activities of antioxidative defense enzymes in the placenta, fetal blood and amnion fluid in inherited thrombophilia. Thrombophilia was associated with nearly threefold increase of activity (p < 0.001 of the placental catalase (81.1 ± 20.6 U/mg of proteins in controls and 270.0 ± 69.9 U/mg in thrombophilic subjects, glutathione (GSH peroxidase (C: 20.2 ± 10.1 U/mg; T: 60.0 ± 15.5 U/mg, and GSH reductase (C: 28.9 ± 5.6 U/mg; T: 72.7 ± 23.0 U/mg. The placental activities of superoxide dismutating enzymes—MnSOD and CuZnSOD, did not differ in controls and thrombophilia. Likewise, the activities of catalase and SOD in the fetal blood, and the level of ascorbyl radical which represents a marker of oxidative status of amniotic fluid, were similar in controls and thrombophilic subjects. From this we concluded that in thrombophilia, placental tissue is exposed to H2O2-mediated oxidative stress, which could be initiated by pro-thrombic conditions in maternal blood. Increased activity of placental H2O2-removing enzymes protects fetus and mother during pregnancy, but may increase the risk of postpartum thrombosis.

  16. Regulation of intramembranous absorption and amniotic fluid volume by constituents in fetal sheep urine. (United States)

    Anderson, Debra F; Jonker, Sonnet S; Louey, Samantha; Cheung, Cecilia Y; Brace, Robert A


    Our objective was to test the hypothesis that fetal urine contains a substance(s) that regulates amniotic fluid volume by altering the rate of intramembranous absorption of amniotic fluid. In late gestation ovine fetuses, amniotic fluid volumes, urine, and lung liquid production rates, swallowed volumes and intramembranous volume and solute absorption rates were measured over 2-day periods under control conditions and when urine was removed and continuously replaced at an equal rate with exogenous fluid. Intramembranous volume absorption rate decreased by 40% when urine was replaced with lactated Ringer solution or lactated Ringer solution diluted 50% with water. Amniotic fluid volume doubled under both conditions. Analysis of the intramembranous sodium and chloride fluxes suggests that the active but not passive component of intramembranous volume absorption was altered by urine replacement, whereas both active and passive components of solute fluxes were altered. We conclude that fetal urine contains an unidentified substance(s) that stimulates active intramembranous transport of amniotic fluid across the amnion into the underlying fetal vasculature and thereby functions as a regulator of amniotic fluid volume.

  17. Challenges in validating the sterilisation dose for processed human amniotic membranes

    Energy Technology Data Exchange (ETDEWEB)

    Yusof, Norimah [Malaysian Nuclear Agency, Bangi, 43000 Kajang, Selangor (Malaysia)], E-mail:; Hassan, Asnah [Malaysian Nuclear Agency, Bangi, 43000 Kajang, Selangor (Malaysia); Firdaus Abd Rahman, M.N.; Hamid, Suzina A. [National Tissue Bank, Hospital Universiti Sains Malaysia, Kubang Kerian, 16130 Kelantan (Malaysia)


    Most of the tissue banks in the Asia Pacific region have been using ionising radiation at 25 kGy to sterilise human tissues for save clinical usage. Under tissue banking quality system, any dose employed for sterilisation has to be validated and the validation exercise has to be a part of quality document. Tissue grafts, unlike medical items, are not produced in large number per each processing batch and tissues relatively have a different microbial population. A Code of Practice established by the International Atomic Energy Agency (IAEA) in 2004 offers several validation methods using smaller number of samples compared to ISO 11137 (1995), which is meant for medical products. The methods emphasise on bioburden determination, followed by sterility test on samples after they were exposed to verification dose for attaining of sterility assurance level (SAL) of 10{sup -1}. This paper describes our experience in using the IAEA Code of Practice in conducting the validation exercise for substantiating 25 kGy as sterilisation dose for both air-dried amnion and those preserved in 99% glycerol.

  18. Cryopreserved Amniotic Suspension for the Treatment of Knee Osteoarthritis. (United States)

    Vines, Jeremy B; Aliprantis, Antonios O; Gomoll, Andreas H; Farr, Jack


    There are few treatment options for symptomatic knee osteoarthritis (OA). Human amniotic suspension allografts (ASA) have anti-inflammatory and chondroregenerative potential and thus represent a promising treatment strategy. In anticipation of a large, placebo-controlled trial of intra-articular ASA for symptomatic knee OA, an open-label prospective feasibility study was performed. Six patients with Kellgren-Lawrence grades 3 and 4 tibiofemoral knee OA were administered a single intra-articular ASA injection containing cryopreserved particulated human amnion and amniotic fluid cells. Patients were followed for 12 months after treatment. No significant injection reactions were noted. Compared with baseline there were (1) no significant effect of the ASA injection on blood cell counts, lymphocyte subsets, or inflammatory markers and (2) a small, but statistically significant increase in serum IgG and IgE levels. Patient-reported outcomes including International Knee Documentation Committee, Knee Injury and Osteoarthritis Outcome, and Single Assessment Numeric Evaluation scores were collected throughout the study and evaluated for up to 12 months. Overall, this study demonstrates the feasibility of a single intra-articular injection of ASA for the treatment of knee OA and provides the foundation for a large placebo-controlled trial of intra-articular ASA for symptomatic knee OA. PMID:26683979

  19. DNA methylation dynamics in human induced pluripotent stem cells over time.

    Directory of Open Access Journals (Sweden)

    Koichiro Nishino


    Full Text Available Epigenetic reprogramming is a critical event in the generation of induced pluripotent stem cells (iPSCs. Here, we determined the DNA methylation profiles of 22 human iPSC lines derived from five different cell types (human endometrium, placental artery endothelium, amnion, fetal lung fibroblast, and menstrual blood cell and five human embryonic stem cell (ESC lines, and we followed the aberrant methylation sites in iPSCs for up to 42 weeks. The iPSCs exhibited distinct epigenetic differences from ESCs, which were caused by aberrant methylation at early passages. Multiple appearances and then disappearances of random aberrant methylation were detected throughout iPSC reprogramming. Continuous passaging of the iPSCs diminished the differences between iPSCs and ESCs, implying that iPSCs lose the characteristics inherited from the parent cells and adapt to very closely resemble ESCs over time. Human iPSCs were gradually reprogrammed through the "convergence" of aberrant hyper-methylation events that continuously appeared in a de novo manner. This iPS reprogramming consisted of stochastic de novo methylation and selection/fixation of methylation in an environment suitable for ESCs. Taken together, random methylation and convergence are driving forces for long-term reprogramming of iPSCs to ESCs.

  20. Evidence for increased SOX3 dosage as a risk factor for X-linked hypopituitarism and neural tube defects. (United States)

    Bauters, Marijke; Frints, Suzanna G; Van Esch, Hilde; Spruijt, Liesbeth; Baldewijns, Marcella M; de Die-Smulders, Christine E M; Fryns, Jean-Pierre; Marynen, Peter; Froyen, Guy


    Genomic duplications of varying lengths at Xq26-q27 involving SOX3 have been described in families with X-linked hypopituitarism. Using array-CGH we detected a 1.1 Mb microduplication at Xq27 in a large family with three males suffering from X-linked hypopituitarism. The duplication was mapped from 138.7 to 139.8 Mb, harboring only two annotated genes, SOX3 and ATP11C, and was shown to be a direct tandem copy number gain. Unexpectedly, the microduplication did not fully segregate with the disease in this family suggesting that SOX3 duplications have variable penetrance for X-linked hypopituitarism. In the same family, a female fetus presenting with a neural tube defect was also shown to carry the SOX3 copy number gain. Since we also demonstrated increased SOX3 mRNA levels in amnion cells derived from an unrelated t(X;22)(q27;q11) female fetus with spina bifida, we propose that increased levels of SOX3 could be a risk factor for neural tube defects.

  1. Expression of natural antimicrobials by human placenta and fetal membranes. (United States)

    King, A E; Paltoo, A; Kelly, R W; Sallenave, J-M; Bocking, A D; Challis, J R G


    Preterm birth associated with infection is a major clinical problem. We hypothesized that this condition is associated with altered expression of natural antimicrobial molecules (beta-defensins (HBD), elafin). Therefore, we examined expression of these molecules and their regulation by proinflammatory cytokines in placentae and fetal membranes from term pregnancy. HBD1-3 and elafin were localized by immunohistochemistry in fetal membranes and placenta. Real-time quantitative PCR was used to examine mRNA expression in primary trophoblast cells treated with inflammatory molecules. HBD1-3 and elafin were immunolocalized to placental and chorion trophoblast layers of fetal membranes and placenta. Immunoreactivity was also observed in amnion epithelium and decidua. No differences were noted between samples from women who were not in labour compared to those in active labour. In in vitro cultures of primary trophoblast cells, HBD2 and elafin mRNA expression was upregulated by the proinflammatory cytokine, IL-1beta. These results suggest that the chorion and placental trophoblast layers may be key barriers to the progression of infection in the pregnant uterus. Natural antimicrobial expression may be altered in response to inflammatory mediator expression associated with the onset of labour and/or uterine infection, providing increased protection when the uterus may be particularly susceptible to infection. PMID:16513165

  2. Target genes of Dpp/BMP signaling pathway revealed by transcriptome profiling in the early D.melanogaster embryo. (United States)

    Dominguez, Calixto; Zuñiga, Alejandro; Hanna, Patricia; Hodar, Christian; Gonzalez, Mauricio; Cambiazo, Verónica


    In the early Drosophila melanogaster embryo, the gene regulatory network controlled by Dpp signaling is involved in the subdivision of dorsal ectoderm into the presumptive dorsal epidermis and amnioserosa. In this work, we aimed to identify new Dpp downstream targets involved in dorsal ectoderm patterning. We used oligonucleotide D. melanogaster microarrays to identify the set of genes that are differential expressed between wild type embryos and embryos that overexpress Dpp (nos-Gal4>UAS-dpp) during early stages of embryo development. By using this approach, we identified 358 genes whose relative abundance significantly increased in response to Dpp overexpression. Among them, we found the entire set of known Dpp target genes that function in dorsal ectoderm patterning (zen, doc, hnt, pnr, ush, tup, and others) in addition to several up-regulated genes of unknown functions. Spatial expression pattern of up-regulated genes in response to Dpp overexpression as well as their opposing transcriptional responses to Dpp loss- and gain-of-function indicated that they are new candidate target genes of Dpp signaling pathway. We further analyse one of the candidate genes, CG13653, which is expressed at the dorsal-most cells of the embryo during a restricted period of time. CG13653 orthologs were not detected in basal lineages of Dipterans, which unlike D. melanogaster develop two extra-embryonic membranes, amnion and serosa. We characterized the enhancer region of CG13653 and revealed that CG13653 is directly regulated by Dpp signaling pathway. PMID:27397649

  3. Prenatal diagnosis and postpartal therapy of fetal obstructions related to the urinary tract. (United States)

    Meyer-Schwickerath, M; Bedow, W; Rascher, W


    114 fetal malformations were diagnosed by ultrasound in the years 1983-1987 at our institute. 32 of these malformations involved the urinary tract. Fetuses with multicystic dysplasic kidneys and oligohydramnios have only a poor prognosis. In obstructive uropathy, however, exact antenatal diagnosis makes early leads to specific postpartal urological therapy. Modern ultrasound equipment makes it possible to localize the site of obstruction and/or dilatation in fetuses. The amount of amnion fluid indicates the degree of obstruction and is an important factor the postpartal prognosis of the children. Relieve of obstruction should be the first step of urological therapy directly after birth. We prefer to perform reconstructive surgery as soon as possible in the first 2 to 3 weeks of life. In children with subpelvic stenosis pyeloplasty is performed in the first weeks of life. Our results of 13 pyeloplasties in newborn are favourable. In children with megaureters dynamic scintigraphy or pressure flow studies (Whitaker test) are performed to diagnose or to exclude obstruction as a cause of dilatation (n = 14). In case of obstruction we perform an ureterocutaneostomy (Ring- or Sober procedure) immediately. Urethral valves causing subvesical obstruction could be treated by transvesical antegrade valve ablation, performed in 9 newborn with good success.

  4. Multilineage Potential Research of Bovine Amniotic Fluid Mesenchymal Stem Cells

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    Yuhua Gao


    Full Text Available The use of amnion and amniotic fluid (AF are abundant sources of mesenchymal stem cells (MSCs that can be harvested at low cost and do not pose ethical conflicts. In human and veterinary research, stem cells derived from these tissues are promising candidates for disease treatment, specifically for their plasticity, their reduced immunogenicity, and high anti-inflammatory potential. This work aimed to obtain and characterize bovine amniotic fluid mesenchymal stem cells (AFMSC. The bovine AF from the amniotic cavity of pregnant gilts in the early stages of gestation (3- and 4-m-old bovine embryos was collected. AFMSCs exhibit a fibroblastic-like morphology only starting from the fourth passage, being heterogeneous during the primary culture. Immunofluorescence results showed that AFMSCs were positive for β-integrin, CD44, CD73 and CD166, but negative for CD34, CD45. Meanwhile, AFMSCs expressed ES cell markers, such as Oct4, and when appropriately induced, are capable of differentiating into ectodermal and mesodermal lineages. This study reinforces the emerging importance of these cells as ideal tools in veterinary medicine; future studies aimed at a deeper evaluation of their immunological properties will allow a better understanding of their role in cellular therapy.

  5. The urticating setae of Ochrogaster lunifer, an Australian processionary caterpillar of veterinary importance. (United States)

    Perkins, L E; Zalucki, M P; Perkins, N R; Cawdell-Smith, A J; Todhunter, K H; Bryden, W L; Cribb, B W


    The bag-shelter moth, Ochrogaster lunifer Herrich-Schaffer (Lepidoptera: Notodontidae), is associated with a condition called equine amnionitis and fetal loss (EAFL) on horse farms in Australia. Setal fragments from O. lunifer larvae have been identified in the placentas of experimentally aborted fetuses and their dams, and in clinical abortions. The gregarious larvae build silken nests in which large numbers cohabit over spring, summer and autumn. The final instars disperse to pupation sites in the ground where they overwinter. Field-collected O. lunifer larvae, their nests and nearby soil were examined using light and electron microscopy to identify setae likely to cause EAFL and to determine where and how many were present. Microtrichia, barbed hairs and true setae were found on the exoskeletons of the larvae. True setae matching the majority of setal fragments described from equine tissue were found on third to eighth instar larvae or exuviae. The number of true setae increased with the age of the larva; eighth instars carried around 2.0-2.5 million true setae. The exuvia of the pre-pupal instar was incorporated into the pupal chamber. The major sources of setae are likely to be nests, dispersing pre-pupal larvae and their exuviae, and pupal chambers. PMID:26669823

  6. Phenotypical Reversion and Differentiation of Breast Cancer in vitro by Sodium Phenylacetate

    Institute of Scientific and Technical Information of China (English)


    Objective To evaluate the effect of a nontoxic differentiation inducer, sodium phenylacetate (NaPA) on breast cancer MCF-7 and MDA-453 cells in vitro. Methods The anchorage growth assay, ELISA, immunofluorescence analysis and electron microscopy were used to study the effect of NaPA on MCF-7 and MDA-453 cells. Results NaPA treatment resulted in a dose-dependent inhibition of MCF-7 or MDA-453 cell growth, including anchorage-dependent and anchorage-independent growth, but il did not significantly inhibit the growth of normal amnion tissue Wish cells and adult hepatic cells L-02. In addition, NaPA could enhance the expressions of intercellular adhesion molecule-1(ICAM-1), human leukocyte antigen ( HLA ) class Ⅰ and Ⅱ molecules on the surface of MCF-7 and MDA-453 cells. Electron micrographs demonstrated richer rough endoplasmic reticulum,mitochondria in NaPA-treated MCF-7 and MDA-453 cells, in contrast to the scant before-mentwned structures but scattered cytoplasmic polyribosomes in the untreated ones. Conclusion NaPA is effective in inducing phenotype reversion and differentiation of breast cancer, and potential in cancer intervention.

  7. The timing of monozygotic twinning: a criticism of the common model. (United States)

    Herranz, Gonzalo


    In the dominant model, monozygotic (MZ) twinning is universally accepted as a post-fertilization event resulting from splitting of the embryo along its first 2 weeks of development. The stage at which splitting occurs determines chorionicity and amnionicity. A short history on how the model was built is presented, stressing the role played by some embryologists, in particular George Corner, in its completion and final success. Strikingly, for more than 60 years no deep criticisms have been raised against the model, which, in virtue of its rational and plausible character, enjoys the status of undisputed truth. At close examination, the embryological support of the model shows some important weak points, particularly when dealing with late splitting. In the author's view, the model not only has contributed to 'suspend' our knowledge on the timing of MZ twinning, but seems indefensible and claims to be substituted. That factor could imply relevant consequences for embryology and bioethics. As an alternative to the model, a new theory to explain the timing of MZ twinning is proposed. It is based on two premises. First, MZ twinning would be a fertilization event. In that case, due to an alteration of the zygote-blastomere transition, the first zygotic division, instead of producing two blastomeres, generates twin zygotes. Second, monochorionicity and monoamnionicity would not depend on embryo splitting, but on fusion of membranes. Some support for this theory can be found in recent embryological advances and also in some explanations of old.

  8. The Consequences of Chorioamnionitis: Preterm Birth and Effects on Development

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    Robert Galinsky


    Full Text Available Preterm birth is a major cause of perinatal mortality and long-term morbidity. Chorioamnionitis is a common cause of preterm birth. Clinical chorioamnionitis, characterised by maternal fever, leukocytosis, tachycardia, uterine tenderness, and preterm rupture of membranes, is less common than subclinical/histologic chorioamnionitis, which is asymptomatic and defined by inflammation of the chorion, amnion, and placenta. Chorioamnionitis is often associated with a fetal inflammatory response. The fetal inflammatory response syndrome (FIRS is defined by increased systemic inflammatory cytokine concentrations, funisitis, and fetal vasculitis. Clinical and epidemiological studies have demonstrated that FIRS leads to poor cardiorespiratory, neurological, and renal outcomes. These observations are further supported by experimental studies that have improved our understanding of the mechanisms responsible for these outcomes. This paper outlines clinical and experimental studies that have improved our current understanding of the mechanisms responsible for chorioamnionitis-induced preterm birth and explores the cellular and physiological mechanisms underlying poor cardiorespiratory, neural, retinal, and renal outcomes observed in preterm infants exposed to chorioamnionitis.

  9. Preparation of denatured protein bone sterilized with gamma radiation; Preparacion de hueso desproteinizado esterilizado con radiacion gamma

    Energy Technology Data Exchange (ETDEWEB)

    Luna Z, D. [ININ, 52045 Ocoyoacac, Estado de Mexico (Mexico)]. e-mail:


    The bone is one of the tissues more transplanted in the entire world by that the bone necessity for transplant every day becomes bigger. In the Bank of tissues Radio sterilized of the ININ the amnion and the pig skin are routinely processed. The tissue with which will be continued is with bone. Due to that in our country it doesn't have enough bone of human origin for the necessities required in the bone transplant, an option is the bone of bovine. Of this bone one can obtain denatured protein bone, with the same characteristics of the denatured protein human bone, the one which has been proven that it has good acceptance and incorporation in the human body when is transplanted. The method for the obtaining of the denatured protein bone of bovine, with the confirmation of the final product by means of X-ray diffraction is described. The radiosterilization of this bone with gamma rays and the determination of the lead content. (Author)

  10. Clinical effect of contact lens treated superficial fungal corneal ulcer after operation%表浅真菌性角膜溃疡术后应用治疗性接触镜临床疗效观察

    Institute of Scientific and Technical Information of China (English)

    于静; 高明宏; 杜春光; 刘乙楠; 刘英


    目的 观察表浅真菌性角膜溃疡行病灶切除联合冷凝及羊膜覆盖手术,术后辅助应用治疗性角膜接触镜的临床效果.方法 临床病例对照研究.对2012年5月至2014年1月在沈阳军区总医院眼科确诊的表浅真菌性角膜溃疡患者93例(93只眼),均采用角膜病灶切除联合冷凝及羊膜覆盖术,术后按数字单偶法随机分为戴镜组(48只眼),给予配戴角膜接触镜;对照组(45只眼)为不配戴角膜接触镜.戴镜时间1~3个月,角膜接触镜每21天更换一次.术后随访时间2~6个月.结果 戴镜组临床总有效率87.50% (42/48),显著高于对照组66.67% (30/45),差异有统计学意义(x2=5.77,P <0.05).戴镜组角膜溃疡治愈时间为(16±2)d,显著少于对照组(24±3)d,差异有统计学意义(t =15.03,P<0.05).结论 表浅真菌性角膜溃疡患者术后连续配戴软性角膜接触镜可以改善临床症状,促进角膜溃疡愈合,提高临床疗效.%Objective To observe the clinical effect of auxiliary application of contact lens in superficial fungal corneal ulcer treated with lesion keratectomy combined with cryotherapy and biological amnion covering.Methods The 93 patients(93 eyes)that had been diagnosed with fungal comeal ulcer were treated with lesion keratectomy combined with cryotherapy and biological amnion covering in our hospital from May 2012 to January 2013.Those were divided into contact lens group (48 eyes)and control group(without contact lens group)(45 eyes)at random.The contact lens group was treated with wearing contact lens for 1 month to 3 months and changing them every 21 days.All the cases were followed up for 2 to 6 months.Results The effective rate in the contact lens group(87.50%)was significantly higher than those in the control group (66.67%) (P <0.05).The cure time of corneal ulcer in the contact lens(16±2 days),was significantly shorter than control group(24± 3 days)(P <0.05).Conclusions Wearing soft contact lens


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    Full Text Available 3986 files have been examined in the Advisory Institute for a five year period in relation to the present risk factors in the pre, peri and postnatal period, the occurrence of asphyxia, I.H. (intracranial hemorrhages and brain edema and their outcome for the children. There were 958 or 32% risk children, out of them 206 or 22% were with asphyxia, 25 or 3% were with brain edema and 14 or 1,5% were with intracranial hemorrhages.The analysis for the risk factors shows that 119 of them were abortive , and from them 15% were born with asphyxia; 124 were SFD and 21% of them with asphyxia; 272 children weighed over 4500 gr., 7% of them with asphyxia and 0.4% with I.H., there were 68 twins, 12% of them with asphyxia. Out of the children with no risk registered, 6 were born with I.H., or 0,2%.Mothers under the age of 18 gave birth to 13% children with asphyxia; treated for sterility and anemia during pregnancy 15%; with increased blood pressure 14%; and 5% with maintained pregnancy.The highest delivery risk is present with children born with vacuum extraction (30% or every third child is with asphyxia and 3% with I.H. and with children delivered by caesarean section (14% with asphyxia.As for the position of the fetus-Citus pedalicus gave 55% children with asphyxia, and Situs pelvicus 12%.The worst damage is suffered by infants with premature amnion disruption (62% are with asphyxia; with the umbilical cord round the neck-56% with asphyxia and 6% with I.H.; and with muddled amniotic fluid and placenta pelvia-50%.The order of risk factors related to asphyxia, I.H. and brain edema is as follows: the first is premature amnion disruption, then follows the umbilical cord round the neck, the muddled amniotic fluid, and placenta previa and Citus pedalicus-which are obstetric problems. The next are the vacuum extraction and S.C. As for the gestatory period the order is as follows: first the abortive, then the twins and hypertrofic infants. The outcome of the

  12. Prostaglandin dehydrogenase and the initiation of labor. (United States)

    Challis, J R; Patel, F A; Pomini, F


    concentrations. Since glucocorticoids appear also to increase expression of prostaglandin synthesizing enzymes within the amnion and chorion, directly by upregulating PGHS-2, or indirectly through the intermediary action of a paracrine effector such as CRH, their role in coordinating processes of parturition remains central. Further understanding of the regulation of PGDH may be of therapeutic importance. For example, it is possible that PGDH activity in lower segment chorion may be reduced in those patients with premature cervical softening, or may be particularly high in those patients with an unfavorable cervix, presenting with a low Bishop score and poor progression at the time of labor. If the enzyme in this region crucially determines the passage and availability of biologically active prostaglandins from amnion and chorion to underlying cervix, then pharmacologic manipulation of PGDH activity may effectively regulate PG transfer in these clinical conditions. Glucocorticoids appear to have a central role in promoting production of agents that are uterotonic to myometrial activity. It is likely that these activities explain the transient increments in uterine contractility reported in patients receiving prenatal corticosteroids to promote fetal pulmonary maturity [11]. Recognition of this physiology suggests that careful monitoring of these patients is advised, and would argue further against repeated, indiscriminate, use of glucocorticoids in patients with an inappropriate diagnosis of threatened preterm labor. PMID:10343931

  13. 人脐血间充质干细胞修复大鼠坐骨神经损伤的实验研究%Experiment research of the function of human umbilical cord blood mesenchymal stem cells in the regeneration of rat's sciatic nerve

    Institute of Scientific and Technical Information of China (English)

    王忠仁; 杨波; 王利民; 李恩


    Objective To evaluate the efectiveness of using mesenchymal stem cells(MSCs)derived from HUCB(human umbilical cord blood)to a tissue engineered bioartificial nerve on bridging a 1 0 mm sciatic nerve gap.Methods The cord blood mononuclear cells were isolated by lymphocyte separation medium,purified and expanded with MesencultTM medium and acidic environment to produce adherent layer(MSCs).Thirty SD female rats were randomly divided into three groups.Group A:Human amnion tubes were seeded by HUCBMSCs together with fibrin sealant.Group B:Human amnion tubes were seeded only with fibrin C:autografts.9 weeks later,a series of examinations were performed which included morphological,sciatic nerve function index,weight of gastrocnemius,histological staining and immunostaining of S100.Results The HUCB-derived mononuclear cells,when seeded in specific medium,gave rise to adherent cells (MSCs).At 9 weeks after the operations,all the examinations of group A was better than group B(P<0.05).Conclusion HUCBMSCs can be isolated,purified, cultivated and expanded Mesencult~(TM) medium.HUCBMSCs can promote the nerve to regenerate in reparing the sciatic nerve gap.%目的 评价用人脐血间充质干细胞(HUCBMSCs)构建组织工程化人工神经修复大鼠坐骨神经10 mm缺损的治疗效果. 方法 用淋巴细胞分离液分离脐血的单个核细胞,以偏酸性的MesencultTM进行培养获得MSCs.30只SD大鼠随机分为3组,每组10只.A组:将HUCBMSCs与生物蛋白胶混合,种植于羊膜管中修复坐骨神经缺损;B组:仅将生物蛋白胶种植于羊膜管中:C组:坐骨神经切下后再将其缝合.9周后,行大体观察、坐骨神经功能指数、腓肠肌湿重测定、组织学染色,S100免疫组化染色等检查. 结果 32份脐血18份可培养出MSCs,但传代培养大量扩增只有4份.HUCBMSCs植人手术后9周检查结果显示,坐骨神经功能指数、腓肠肌湿重测定A组(-64.2234±2.9461、41.29524±3.88421)

  14. 组织工程上皮移植对碱烧伤角膜新生血管的影响%Tissue-engineered epithelium transplantation on corneal neovascularization in alkali-induced limbal stem cell deficiency

    Institute of Scientific and Technical Information of China (English)

    郭青; 皮裕琍


    目的:评价组织工程上皮移植在碱烧伤角膜缘干细胞缺乏症中对角膜新生血管的抑制作用.方法:回顾性非随机的病例研究.2006/2011年我院收治的19例(23眼)完全性角膜缘干细胞缺乏症的碱烧伤患者,10例13眼行组织工程上皮移植,9例10眼行羊膜移植.所有患者在手术前后均用裂隙灯观察角膜新生血管情况,在术后第21,60d对角膜新生血管进行评分比较.结果:术后第21d和术后第60d组织工程上皮移植组和羊膜移植组角膜新生血管均较术前明显减少( P<0.05),在术后两个评价时间点,组织工程上皮移植组平均角膜新生血管分数明显低于羊膜移植组.结论:对碱烧伤所致角膜缘干细胞缺乏的患者,组织工程上皮移植抑制角膜新生血管的作用明显好于羊膜移植.%AIM: To evaluate the clinical outcomes of tissue-engineered epithelium transplantation on decreasing corneal neovascularization in patients with limbal stem cell deficiencies caused by alkali burns.eyes of 19 cases with severe corneal alkali burns from 2006 to 2011 in our hospital. All of these eyes had complete limbal stem cell deficiencies. Ten cases (13 eyes) were performed with tissue-engineered epithelium transplantation, and 9 cases (10 eyes) were performed with amnion membrane transplantation. Vascularization was observed in all cases by slit-lamp microscope before and after surgery. A neovascularization scoring system was used to evaluate each eye at the 21th and 60th after surgery.transplantation or amniotic membrane transplantation was performed, corneal neovascularization decreased significantly (P<0.05). And averaged scores of corneal neovascularization were significantly lower in tissue-engineered epithelium transplantation group at the 21th day and 60th day after surgery than those of amniotic membrane transplantation group(P<0.05). inhibitting the growth of corneal neovascularization in patients with limbal stem cell deficiencies

  15. Prenatal diagnosis of thalassemia and hemoglobinopathies in Thailand: experience from 100 pregnancies. (United States)

    Fucharoen, S; Winichagoon, P; Thonglairoam, V; Siriboon, W; Siritanaratkul, N; Kanokpongsakdi, S; Vantanasiri, C


    In this review, we describe a simple strategy to detect the three severe thalassemic diseases commonly found in Thailand. Hb Bart's hydrops fetalis can be detected unambiguously by ultrasonography at 18-20 weeks of gestation or detected early in the first trimester by the gene amplification technique. Prenatal diagnosis for homozygous beta-thalassemia is better performed in the second trimester by in vitro protein synthesis. This is because the molecular defects of some beta-thalassemias are still unknown and homozygosity of the same mutation is low. In contrast, beta-thalassemia/Hb E is easily detected, in the first trimester, by direct visualization on electrophoresis or by dot blot analysis of enzymatically amplified DNA with a set of nonradioactively labeled oligonucleotide probes complementary to the most common mutations. We also found that the beta/gamma synthesis ratio in homozygous Hb E is similar to that of beta-thalassemia/Hb E and DNA analysis is the only method to distinguish these two conditions in the couple at risk of having either beta-thalassemia/Hb E or asymptomatic homozygous Hb E. In 100 pregnancies studied, the diagnoses were achieved in 96 pregnancies. Complications leading to fetal loss were found in 3 pregnancies: one woman developed amnionitis after fetal blood sampling; one had amniotic fluid leakage after the biopsy, and the third, carrying a normal fetus, aborted 10 days after fetal blood sampling with urinary tract infection and high fever. However, these figures are compatible with other reports and the risks are significantly lower than that of thalassemic disease the fetus is facing. One case of beta-thalassemia/Hb E was incorrectly diagnosed prenatally as being Hb E trait. In twenty-five pregnancies (25%) prenatally diagnosed to carry affected fetuses it was decided to have abortion. This study shows the feasibility of prenatal diagnosis for thalassemic diseases in Thailand which, in addition to screening and genetic counseling

  16. Evaluation of antiviral activity of fractionated extracts of sage Salvia officinalis L. (Lamiaceae

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    Šmidling Dragana


    Full Text Available In the present study, we examined cytotoxicity and extracellular and intracellular antiviral activity of frac­tionated extracts of wild and cultivated sage Salvia officinalis L. (Lamiaceae in vitro using the WISH-VSV model system. Extracts were obtained by fractionating depigmented ethanol extracts of sage plants with supercritical CO2 at different pressures. Cytotoxicity was determined by examining cellular morphology in situ with the aid of a colorimetric micromethod and by cell staining with trypan blue. The fraction of distilled cultivated sage obtained at CO2 pressure of 300 bars and temperature of 60°C (149/3 was the most cytotoxic, with CTD10 44 μg/ml. That of non-distilled cultivated sage obtained at CO2 pressure of 500 bars and temperature of 100°C (144/5 was the least toxic (CTD10 199 μg/ml. Moreover, 144/5 had an antiviral effect at the intracellular level: when added 5 hours before VSV infection, it caused 100% reduction of CPE at concentrations of 99.5 and 199.0 μg/ml; when added after virus penetration had occurred, the same concentrations caused 35 and 60% reduction, respectively. The obtained results indicate that antiviral activity of 144/5 involves inhibition of the early steps of the virus infective cycle without a direct virucidal effect. Abbreviations: WISH - human amnion epithelial cells, VSV - vesicular stomatitis virus, HSV - herpes simplex virus, CPE - cytopathic effect, IS - selectivity index, TCID50 - tissue culture infective dose, CTD10 - 10% cytotoxic concentrations.

  17. Expression of the prostaglandin F synthase AKR1B1 and the prostaglandin transporter SLCO2A1 in human fetal membranes in relation to spontaneous term and preterm labour

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    Hana A Alzamil


    Full Text Available Background: Human labour is a complex series of cellular and molecular events that occur at the materno-fetal and uterine levels. Many hypotheses have been proposed for the initiation of human labour, one hypothesis suggests that maturation of the fetus releases a signal in the amniotic fluid that will be transmitted to myometrium via the fetal membranes and initiate uterine contractions. There is strong evidence that prostaglandins (PGs play a central role in initiation and progression of human labour. Objectives: In this study we intended to investigate the expression of prostaglandin F synthase and the prostaglandin transporter in the human fetal membranes and to explore the relationship between cytokines and PGs in the mechanism of human labour. Methods: We used fetal membranes obtained before labour at term and after spontaneous labour at term or preterm to identify the changes in prostaglandin F synthase (AKR1B1 and human prostaglandin transporter (SLCO2A1 proteins in relation to parturition. Using fetal membranes explants we tested the effect of cytokines (interleukin-1 and tumour necrosis factor alpha on PG production and the concomitant changes in cyclooxygenase-2 (PTGS2, AKR1B1 and SLCO2A1 expression. Results: Expression of PTGS2 and AKR1B1 was upregulated in the fetal membranes in association with term labour while SLCO2A1 was downregulated with advancing gestation and during term labour. Before labour, IL-1 increased the expression of PTGS2, however during labour TNF upregulated PTGS2 and AKR1B1 proteins. Conclusions: The prostaglandin F synthase AKR1B1 is upregulated while prostaglandin transporter is downregulated during term labour. The amnion is more responsive than choriodecidua to stimulation with pro-inflammatory cytokines. The mechanisms of term and preterm labour are different.

  18. Mesenchymal stem cells from rat olfactory bulbs can differentiate into cells with cardiomyocyte characteristics. (United States)

    Huang, Yuahn-Sieh; Li, I-Hsun; Chueh, Sheau-Huei; Hueng, Dueng-Yuan; Tai, Ming-Cheng; Liang, Chang-Min; Lien, Shiu-Bii; Sytwu, Huey-Kang; Ma, Kuo-Hsing


    Mesenchymal stromal/stem cells (MSCs) are widely distributed in different tissues such as bone marrow, adipose tissues, peripheral blood, umbilical cord and amnionic fluid. Recently, MSC-like cells were also found to exist in rat olfactory bulb and are capable of inducing differentiation into mesenchymal lineages - osteocytes, chondrocytes and adipocytes. However, whether these cells can differentiate into myocardial cells is not known. In this study, we examined whether olfactory bulb-derived MSCs could differentiate into myocardial cells in vitro. Fibroblast-like cells isolated from the olfactory bulb of neonatal rats were grown under four conditions: no treatment; in the presence of growth factors (neuregulin-1, bFGF and forskolin); co-cultured with cardiomyocytes; and co-cultured with cardiomyocytes plus neuregulin-1, bFGF and forskolin. Cell differentiation into myocardial cells was monitored by RT-PCR, light microscopy immunofluorescence, western blot analysis and contractile response to pharmacological treatments. The isolated olfactory bulb-derived fibroblast-like cells expressed CD29, CD44, CD90, CD105, CD166 but not CD34 and CD45, consistent with the characteristics of MSCs. Long cylindical cells that spontaneously contracted were only observed following 7 days of co-culture of MSCs with rat cardiomyocytes plus neuregulin-1, bFGF and forskolin. RT-PCR and western blot analysis indicated that the cylindrical cells expressed myocardial markers, such as Nkx2.5, GATA4, sarcomeric α-actinin, cardiac troponin I, cardiac myosin heavy chain, atrial natriuretic peptide and connexin 43. They also contained sarcomeres and gap junction and were sensitive to pharmacological treatments (adrenal and cholinergic agonists and antagonists). These findings indicate that rat olfactory bulb-derived fibroblast-like cells with MSC characteristics can differentiate into myocardial-like cells.

  19. The effects of cryopreservation on angiogenesis modulation activity of human amniotic membrane. (United States)

    Yazdanpanah, Ghasem; Paeini-Vayghan, Ghodsieh; Asadi, Samira; Niknejad, Hassan


    Amniotic membrane (AM), as the innermost layer of placenta, has side dependent effects on the angiogenesis. Cryopreservation is a necessary process to avoid the challenging problems of fresh tissues; a procedure which makes the AM ready-to-use. Since the cryopreservation can influence the AM characteristics for experimental and clinical purposes, in this study the effects of cryopreservation were evaluated on angiogenesis modulation activity of the AM compared to fresh tissues in an animal model. The AM was implanted mesenchymal side up or epithelial side up in a rat dorsal skinfold chamber. The length and number of branches of formed capillaries were measured via intravital microscopy after 7 days. The amount of IL-8 (interleukin-8) and TIMP-2 (Tissue Inhibitor of Matrix Metalloproteinase-2) as two factors in amniotic cells which have great impacts on angiogenesis were evaluated using ELISA assay. The epithelial surface of cryopreserved AM had inhibitory effects on vessel formation. The cryopreserved amniotic mesenchymal side increased the vessel length and sprout. The result of cryopreserved AM on angiogenesis was similar to that of fresh tissues. The levels of IL-8 and TIMP-2 in cryopreserved samples were significantly less than fresh AMs which shows that angio-modulatory properties are not limited to the effects of amnion epithelial and mesenchymal stem cells and the other components such as extracellular matrix may contribute in angio-modulatory effects. These promising results show that inducing and inhibitory effects of the AM, which make it an appropriate candidate for different clinical situations, were maintained after cryopreservation.

  20. 成体干细胞向血管内皮细胞定向分化的研究现状%Oriented differentiation of adult stem cells into vascular endothelial cells

    Institute of Scientific and Technical Information of China (English)

    宋秀军; 江其生


    Adult stem cells identified in tissue organs possess obviously plasticity. They can differentiate into the cells in other systems and germ layers, which are known to support the continuous repair and regeneration of tissues. In the vascular tissue engineering, the research was more and more widespread and deep on adult stem cells as seed cells~ Research indicated that there are many kinds of adult stem cells which can differentiate into vascular endothelial cells, such as bone marrow mesenchymal stem cells, hematopoietic stem cells, human adipose derived stem cells, and amnion derived stem calls. Thus, this study reviewed the current situation of adult stem cells as seed cells in vascular tissue engineering.%成体干细胞存在于已分化的组织器官中,具有明显的可塑性,能跨系统、跨胚层分化,在组织工程和损伤修复领域具有重要的应用价值.在血管组织工程研究领域中,成体干细胞作为种子细胞得到越来越广泛而深入的研究.目前研究表明,具有向血管内皮细胞分化的成体干细胞主要有骨髓间充质干细胞、造血干细胞、脂肪来源的干细胞以及羊膜来源的干细胞等基于此,就成体干细胞作为种子细胞在血管组织工程领域研究现状进行综述.

  1. Upregulated expression of Ezrin and invasive phenotype in malignantly transformed esophageal epithelial cells

    Institute of Scientific and Technical Information of China (English)

    Zhong-Ying Shen; Li-Yan Xu; Ming-Hua Chen; En-Min Li; Jin-Tao Li; Xian-Ying Wu; Yi Zeng


    AIM: To investigate the correlation between ezrin expression and invasive phenotype formation in malignantly transformed esophageal epithelial cells. METHODS: The experimental cell line employed in the present study was originated form the progressive induction of a human embryonic esophageal epithelial cell line (SHEE)by the E6E7 genes of human papillomavirus (HPV) type 18.The cells at the 35th passage after induction called SHEEIMM were in a state of immortalized phase and used as the control,while that of the 85th passage denominated as SHEEMT represented the status of cells that were malignantly transformed. The expression changes of ezrin and its mRNA in both cell passages were respectively analyzed by RT-PCR and Western blot. Invasive phenotype was assessed in vivo by inoculating these cells into the severe combined immunodeficient (SCID) mice via subcutaneous and intraperitoneal injection, and in vitro by inoculating them on the surface of the amnion membranes, which then was determined by light microscopy and scanning electron microscopy. RESULTS: Upregulated expression of ezrin protein and its mRNA was observed in SHEEMT compared with that in SHEEIMM cells. The SHEEMT cells inoculated in SCID mice were observed forming tumor masses in both visceral organs and soft tissues in a period of 40 days with a special propensity to invading mesentery and pancreas, but did not exhibit hepatic metastases. Pathologically, these tumor cells harboring larger nucleus, nucleolus and less cytoplasm could infiltrate and destroy adjacent tissues. In the in vitro study,the inoculated SHEEMT cells could grow in cluster on the amniotic epithelial surface and intrude into the amniotic stroma. In contrast, unrestricted growth and invasiveness were not found in SHEEIMM cells in both in vivo and in vitroexperiment. CONCLUSION: The upregulated ezrin expression is one of the important factors that are possibly associated with the invasive phenotype formation in malignantly

  2. Photographic-Based Optical Evaluation of Tissues and Biomaterials Used for Corneal Surface Repair: A New Easy-Applied Method.

    Directory of Open Access Journals (Sweden)

    Miguel Gonzalez-Andrades

    Full Text Available Tissues and biomaterials used for corneal surface repair require fulfilling specific optical standards prior to implantation in the patient. However, there is not a feasible evaluation method to be applied in clinical or Good Manufacturing Practice settings. In this study, we describe and assess an innovative easy-applied photographic-based method (PBM for measuring functional optical blurring and transparency in corneal surface grafts.Plastic compressed collagen scaffolds (PCCS and multilayered amniotic membranes (AM samples were optically and histologically evaluated. Transparency and image blurring measures were obtained by PBM, analyzing photographic images of a standardized band pattern taken through the samples. These measures were compared and correlated to those obtained applying the Inverse Adding-Doubling (IAD technique, which is the gold standard method.All the samples used for optical evaluation by PBM or IAD were histological suitable. PCCS samples presented transmittance values higher than 60%, values that increased with increasing wavelength as determined by IAD. The PBM indicated that PCCS had a transparency ratio (TR value of 80.3 ± 2.8%, with a blurring index (BI of 50.6 ± 4.2%. TR and BI obtained from the PBM showed a high correlation (ρ>|0.6| with the diffuse transmittance and the diffuse reflectance, both determined using the IAD (p<0.005. The AM optical properties showed that there was a largely linear relationship between the blurring and the number of amnion layers, with more layers producing greater blurring.This innovative proposed method represents an easy-applied technique for evaluating transparency and blurriness of tissues and biomaterials used for corneal surface repair.

  3. Comparative Studies of the Chromosomal Arrangement in the C-Metaphase Between Normal Karyotype and Trisomy-21

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    D.D. Farhud


    Full Text Available Human chromosomes in amnion cells and lymphocytes with normal karyotype and in lymphocytes with pathological karyotype (2n=47, +21 were compared as to their position in the metaphase. None of the collectives showed sex differences. Measurement of the radial distances revealed more peripheral position of the majority of large chromosomes. The satellite-carrying chromosomes of the D group always had a central position in the mitosis. The chromosomes of the groups D, E, F and G were closest to the centre; with the exception of chromosome 18 which was peripheral in all three collectives. For the male probands, the y-chromosome was shown in all three collectives to have a smaller radial distance than the x-chromosome. A typical distribution was found for the radial and homologue distances for the trisomic cells, two of them had a very large radial distance, the third a value corresponding to its size. For the homolarger measurements hereby the distribution is quite independent of parental source. Comparison of the groups showed no differences either between normal and trisomy cells or between the different cell types. Examination of chromosomes 6 and 15 proved conclusively that the chromosomes are not particularly orientated in the c-metaphase regarding the position of short and long arm. A preferential combination of particular satellite carrying chromosomes leads to the frequent fusions of chromosomes 13 and 14, or 14 and 21. Equally, no preferential association could be demonstrated of the chromosome 21 and the chromosomes with large heterochromatin blocks in the centromere region (chromosomes 1 and 9. The distances were of the same order of magnitude as those between 21 and chromosome 6, a submetacentric chromosome without a marked heterochromatin region. Both latter observations are of specific importance for genetic councelling of couples after birth of a child with a de Novo chromosome aberration asking for the recurrence risk.

  4. Maternal Serum Serpin B7 Is Associated With Early Spontaneous Preterm Birth (United States)

    Parry, Samuel; Zhang, Heping; Biggio, Joseph; Bukowski, Radek; Varner, Michael; Xu, Yaji; Andrews, William W.; Saade, George R.; Esplin, M. Sean; Leite, Rita; Ilekis, John; Reddy, Uma M.; Sadovsky, Yoel; Blair, Ian A.


    Objective To identify serum biomarkers of early spontaneous preterm birth (SPTB) using semi-quantitative proteomic analyses. Study Design Nested case-control study of pregnant women with previous SPTB. Maternal serum was collected at 19 to 24 and 28 to 32 weeks gestation, and analyzed by liquid chromatography-multiple-reaction monitoring-mass spectrometry. Targeted and shotgun proteomics identified 31 candidate proteins that were differentially expressed in pooled serum samples from spontaneous preterm (<34 weeks - cases) and term deliveries (controls). Candidate protein expression was compared in individual serum samples between cases and controls matched by age and race groups, and clinical site. Protein expression was verified by Western blot in the placenta and fetal membranes from cases and controls. Results Serum samples were available for 35 cases and 35 controls at 19 to 24 weeks, and 16 cases and 16 controls at 28 to 32 weeks. One protein, serpin B7, yielded serum concentrations that differed between cases and controls. The mean concentration of serpin B7 at 28 to 32 weeks was 1.5-fold higher in women with subsequent preterm deliveries compared to controls; there was no difference at 19 to 24 weeks. Higher levels of serpin B7 at both gestational age windows were associated with a shorter interval to delivery, and higher levels of serpin B7 in samples from 28 to 32 weeks were associated with a lower gestational age at delivery. Western blotting identified serpin B7 protein in placenta, amnion, and chorion from cases and controls. Conclusion Targeted and shotgun serum proteomics analyses associated one protein, serpin B7, with early SPTB. Our results require validation in other cohorts and analysis of the possible mechanistic role of serpin B7 in parturition. PMID:24954659

  5. Chemical and thermal injuries of the eyes. Surgical and medical treatment based on clinical and pathophysiological findings. (United States)

    Reim, M; Redbrake, C; Schrage, N


    Light burns heal well within a few days. Severe chemical and thermal injuries of the eyes destroy surface epithelia and cause ischemic necroses of conjunctiva, cornea, sclera, iris, ciliary body, and lids. An inflammatory response follows with leucocyte infiltration and release of inflammatory mediators. Prostaglandins, lipoxygenase products, cytokines, superoxide radicals and Iysosomal enzymes are known to be active in eye burn disease. Their activities result in corneal, scleral and conjunctival ulceration, tissue proliferation and scarification, which develop within weeks, months and even years after the accident. Pathophysiological events produce defined clinical pictures. Some agents take special actions, e.g. alkali penetrates within seconds into the anterior chamber, sulfuric acid burns as well as quick lime burns forming slaked lime produce considerable heat. Hydrofluoric acid is highly toxic and induces early necroses. Heat causes deep ischemic necroses and lateron strongly shrinking scars. Onset and intensity of first aid decided on the outcome. Immediate rinsing is essential. Cool water, saline, Ringers lactate solution and BSS are good rinsing media. For first aid, buffered Previn seems suitable. Major chemical and thermal injuries need a variety of medical and surgical treatments: Necroses must be excised surgically. Tenon plasty is performed to reconstruct conjunctiva. Amnion-, limbus- and early keratoplasty or artificial epithelium are applied, initially to save the cornea from melting, and later to restore vision. Conjunctical, lid and intraocular surgery may be necessary. The aim of medical treatment is to suppress the inflammatory response and to prevent infection. Corticosteroids, antibiotics, ascorbate and inhibitors of proteolytic enzymes are used. Secondary glaucoma must not be forgotten. Extensive therapy is sometimes rewarding, results are presented. PMID:11228610

  6. Comparative Characterization of Cells from the Various Compartments of the Human Umbilical Cord Shows that the Wharton's Jelly Compartment Provides the Best Source of Clinically Utilizable Mesenchymal Stem Cells.

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    Arjunan Subramanian

    Full Text Available The human umbilical cord (UC is an attractive source of mesenchymal stem cells (MSCs with unique advantages over other MSC sources. They have been isolated from different compartments of the UC but there has been no rigorous comparison to identify the compartment with the best clinical utility. We compared the histology, fresh and cultured cell numbers, morphology, proliferation, viability, stemness characteristics and differentiation potential of cells from the amnion (AM, subamnion (SA, perivascular (PV, Wharton's jelly (WJ and mixed cord (MC of five UCs. The WJ occupied the largest area in the UC from which 4.61 ± 0.57 x 106 /cm fresh cells could be isolated without culture compared to AM, SA, PV and MC that required culture. The WJ and PV had significantly lesser CD40+ non-stem cell contaminants (26-27% compared to SA, AM and MC (51-70%. Cells from all compartments were proliferative, expressed the typical MSC-CD, HLA, and ESC markers, telomerase, had normal karyotypes and differentiated into adipocyte, chondrocyte and osteocyte lineages. The cells from WJ showed significantly greater CD24+ and CD108+ numbers and fluorescence intensities that discriminate between MSCs and non-stem cell mesenchymal cells, were negative for the fibroblast-specific and activating-proteins (FSP, FAP and showed greater osteogenic and chondrogenic differentiation potential compared to AM, SA, PV and MC. Cells from the WJ offer the best clinical utility as (i they have less non-stem cell contaminants (ii can be generated in large numbers with minimal culture avoiding changes in phenotype, (iii their derivation is quick and easy to standardize, (iv they are rich in stemness characteristics and (v have high differentiation potential. Our results show that when isolating MSCs from the UC, the WJ should be the preferred compartment, and a standardized method of derivation must be used so as to make meaningful comparisons of data between research groups.

  7. Effect of Intravenous Dexamethasone on Preparing the Cervix and Labor Induction

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    Fatemeh Laloha


    Full Text Available The use of corticosteroids is one of the methods put forward for the strengthening and speeding up the process of labor. After identification of glucocorticoid receptors in human amnion, the role of corticosteroids in starting the process of labor has been studied in numerous studies. The purpose of this study was to determine the effect of intravenous Dexamethasone on preparing the cervix and on labor induction. A randomized, clinical, and double – blind trial was conducted on 172 women divided into a control and an experimental group. The inclusion criteria were that they had to be primparous, in or before the 40th week of pregnancy, and with Bishop scores (B.S.s of 4 or lower. The exclusion criteria were diabetes, preeclampsia, macrosomia, twin pregnancy, rupture of the membrane (ROM, breech, and women suffering from background diseases. The B.S.s of the women was measured in charge of the study, and each woman was intravenously injected with eight milligrams of Dexamethasone or eight milligrams of distilled water. Four hours after the injections, the B.S.s of the participants was measured, and they were put under the conditions of labor induction using oxytocin. Information was collected in checklists A and B. The patients were compared with respect to B.S., the time the induction started, the average interval between the start of induction and the beginning of the active phase of childbirth, and the average length of time between the start of the active phase and the second stage of childbirth. The first and five minutes Apgar scores of the two groups of women were compared. The frequencies, the means,  and the standard deviations were calculated using the SPSS – 16 software, and analysis of the results was performed with the Student’s t- test and the chi-square test with PPPP

  8. Decidual transformation in adenomyosis during pregnancy as an indication for hysterectomy

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    Jovanović Božidar


    Full Text Available Introduction. Incidence of adenomyosis is estimated on 20% in all of extracted uteruses. Clinically, it usually exists asymptomatically, but could be followed with dysmenorrhoea, menorrhagia and pelvic pain. One third of patients with adenomyosis are sterile, however in other two thirds, conception and pregnancy could have normal development, with delivery without complications. One of possible complications related to adenomyosis is rupture of uterus during delivery. Case report. A pregnant woman at the end of the 10th lunar month of pregnancy, showed minimal labour contractions, following amnion rupture and the delivery was terminated by section ceasar. Subtotal hysterectomy was performed because of concominant profuse bleeding. Myometrium was occupied with decidual cells in trabecular arrangement, among which elongated endometrial glands were placed. Surrounding muscle fascicles showed atrophic, apoptic, and occasionally necrotic changes. Myometrial stroma was edematous, and infiltrated with mesenchymal cells, as well as with adipose cells, which were extending toward serosa. Discussion. The patient had been receiving treatment, for a long period, because of sterility without clear etiological factor. A small number of similar cases ends with complications. Here, it is evident that abundant decidual transformation of stromal cells in adenomiosis leads to atrophy and necrosis of muscle cells. Teh reduction of uterine muscle mass causes atony and threating rupture, through separation of muscle cells, and therefore the absence of their synchronized contractions. Conclusion. Atony of uterine muscle could be caused by decidual transformation in adenomyosal fields, atrophy of muscle fascicles, edema of the stroma, mesenchymal transformation, and fibrosis. This leads to a decrease in myometrial contractions, and prolonged postpartal bleeding.

  9. Limb body wall complex or body stalk complex or cyllosomas: a case report

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    S. Saritha


    Full Text Available Limb body wall complex (LBWC is also called Body stalk complex and Cyllosomas. We present this rare congenital malformation complex highlighting the importance of early sonographic imaging findings in LBWC along with differentiation from other anterior abdominal wall defects. Limb body wall complex / Body stalk anomaly refers to a rare complicated polymalformative fetal malformation syndrome of uncertain etiology and results in head, heart, lung, diaphragm, kidney or gonadal abnormalities. LBWC was first described by Van Allen et al; in (1987. The two of the three following anomalies must be present to establish the diagnosis: 1. Exencephaly / Encephalocele with facial clefts, 2. Thoraco-Abdominoschisis / ventral body wall defects and 3. Limb defects. LBWC arises as a result of early amnion disruptions or error in embryonic development. If all components of the syndrome are present, the condition is lethal. LBWC is invariably fatal and incompatible with life. No case of postnatal survival is reported so far. Serum alpha-fetoprotein measurement and ultrasonography examination is the key to the prenatal diagnosis and followed by medical termination of pregnancy. It presents two distinct phenotypes described by Russo et al (1993 and later Cusi et al in (1996, according to the foetoplacental relationships: 1. Placento-cranial and 2.Placento-abdominal types. Among the 168 live births at S.V.S. Medical College & hospital Mahabubnagar (INDIA during the period of 2010-2011 we came across an aborted female fetus. It was weighing 1800gms, 30 weeks of gestation diagnosed by antenatal ultrasonography as ventral body wall defect. It was associated with ompholocele, severe scoliosis and limb defects. Its confirmation of the diagnosis of Limb body wall complex with Placento-abdominal type was done by postmortem fetography. [Int J Res Med Sci 2013; 1(2.000: 132-137

  10. Correlates of vaginal colonization with group B streptococci among pregnant women

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    Tsering Chomu Dechen


    Full Text Available Background: A study of genital colonization by group B streptococcus (GBS was conducted in pregnant women in their third trimester, which is a known risk factor of morbidity and mortality among newborns. Aims: The present study was undertaken to study the prevalence and the correlates of vaginal colonization by GBS among pregnant women. Setting and Design: This observational cross-sectional study was conducted during September 2002 to March 2004 on 524 pregnant women. Materials and Methods: Three high vaginal swabs were obtained from all the pregnant women admitted at term and in preterm labor. Two swabs were used for aerobic culture and the third one for gram staining. The first set of swabs was cultured on 5% Sheep blood agar plates. The second set of swabs were inoculated into Todd-Hewitt broth and then subcultured in 5% Sheep blood agar plates. The main outcome measures were the presence of GBS infection in comparison to the age group, gravida, gestational age, premature rupture of membrane (PROM, preterm labor and association with febrile spells of the present pregnancy. Results: The culture positivity rate of GBS was 4.77% and coexistent organisms isolated were Candida species (36%, Staphylococcus aureus (8% and Enterococcus species (8%. Culture positivity in the age group of 18-25 years was 5.71%, of which 5.74% were in their first pregnancy. The correlation between age group and gravida with GBS culture positivity was statistically insignificant. The culture positivity in <36 weeks of gestational age was 6.93%. This relation was statistically significant. Twenty-eight percent developed PROM. Sixty-four percent of culture positives had preterm labor. Conclusion: GBS infection among pregnant women was significantly correlated with the gestational age, PROM and preterm labor. In pregnancy GBS colonization causes asymptomatic bacteriuria or UTI. It is a well known cause of puerperal infections with amnionitis,endometritis and sepsis being

  11. Surgical tissue banking - the role of nuclear technology

    International Nuclear Information System (INIS)

    Before 1980s amniotic membranes were mainly used clinically either in fresh forms or preserved in glycerol without sterilization. There were problems of graft infections and difficulty in maintaining standard of graft quality and uniformity of the amnion when applied on the recipient patients. Nuclear technology has helped to sterilize these membranes such that they can be processed, dried and then double packed before finally irradiated with gamma rays emitted from the radionuclide cobalt-60. The irradiated amniotic membranes can be kept at room temperature and are readily available for used by doctors without relying on glycerol for preservation or freezers for storage. Similarly allograft bones can be processed and be stored in similar manner otherwise they need to be frozen at -80 to -160 degree C. Radiation technology has benefited many patients in many countries in the Asia Pacific region through sterilization of tissues. Bone such as femoral heads can be processed into radiation sterilized bone chips or grounded bones and has been successfully used for filling up of cavities left by benign bone tumors, bone cysts etc. These materials are safe and cheap and save the recipients in-term of money and a second surgery and on the extreme case, patients may avoid amputation if an irradiated long bone is available. Orthopedic, neuro- and dental surgeons have reported many such successful outcomes. Also bum patients have been shown to benefit from amniotic membranes or porcine skin which were processed and irradiated prior to treatment. In some countries in this region, amniotic membranes have been used as en effective wound dressing for leprosy patients as well. Reports of these cases are to be presented. (Author)

  12. Association of coexisting morphological umbilical cord abnormality and clinical cord compromise with hypoxic and thrombotic placental histology. (United States)

    Stanek, Jerzy


    To assess the usefulness and limitations of placental histology when morphological umbilical cord (UC) abnormality coexists with clinical UC compromise, 5634 consecutive placentas were divided into four groups and statistically compared: group 1-182 placentas from pregnancies with clinical features of UC compromise (variable decelerations, UC entanglement, prolapse, or true knot at delivery); group 2-1355 placentas with abnormal UC morphology or insertion; group 3-152 placentas with at least one phenotype from group 1 and one from group 2; group 4-3945 placentas with no clinical or morphological UC-related phenotypes (control group).Differences were analyzed by ANOVA or χ (2). Of 68 phenotypes studied, 13 clinical and 18 placental phenotypes were statistically significant. In group 1, 2 phenotypes were most common (oligohydramnios and abnormal fetal heart rate tracing). In group 2, 6 phenotypes were most common, including 4 clinical (abnormal umbilical artery Dopplers, nonmacerated stillbirth, multiple pregnancy, and fetal growth restriction) and 2 placental. In group 3, 23 phenotypes were most common, including 7 clinical (gestational hypertension, polyhydramnios, induction of labor, cesarean section, macerated stillbirth, congenital malformations, and abnormal 3rd stage of labor) and 16 placental. The existence of clinical signs of UC compromise alone was associated with the absence of pathomorphological placental abnormalities. However, the coexistence of clinical and abnormal morphological UC phenotypes was statistically significantly associated with placental histological signs of decreased fetal blood flow, hypoxia (acute and chronic post uterine), shallow placental implantation, and/or amnion nodosum. Thus, confirmation of clinical UC compromise should not be expected on placental examination if no morphological UC abnormality or abnormal UC insertion has been found. PMID:26983702

  13. Expression of aquaporin 3 and aquaporin 9 in placenta and fetal membrane with idiopathic polyhydramnios%水通道蛋白3、9在特发性羊水过多产妇胎盘和胎膜中的表达变化及意义

    Institute of Scientific and Technical Information of China (English)

    朱雪琼; 蒋珊珊; 邹双微; 胡迎春; 王玉环


    Objective To investigate the pathogenesis role of aquaporin 3 and aquaporin 9 in idiopathic polyhydramnios by detecting their expression and distribution in fetal membranes and placenta.Methods Twenty-one of term pregnancy women with idiopathic polyhydramnios were enrolled as patient group matched with 30 women with normal term pregnancy as control group.The expression and localization of aquaporin 3 and aquaporin 9 in fetal membranes and placenta were detected by real-time polymerase chain reaction and streptavidin peroxidase immunohistochemiscal staining.Results (1)The mRNA expressions of aquaporin 3 and aquaporin 9 were detected in amnion,chorion and placental tissue in both patient group and control group.Both aquaporin 3 and aquaporin 9 were demonstrated positive staining in the amnion epithelia,chorion cytotrophoblasts and placental trophoblast.(2)The ratio of aquaporin 3 and aquaporin 9 mRNA expressions in amnion in patient group comparing to those in control group were 5.00 and 3.25,while in chorion they were 2.03 and 2.08.When compared with those in amnion and chorion of control group,there was a significant difference(P<0.01).However,the relative change fold of aquaporin 3 and aquaporin 9 in placental trophoblast in patient group were decreased in comparison of those in control group,which also showed statistical difference(P<0.01).(3)The expression of aquaporin 3 and aquaporin 9 protein in anmion were 7.5 ±2. 0 and 11.1 ± 1.8 in patient group, while they were 5.3 ± 1. 6 and 5.6 ± 2. 3 in control group. In chorion, the expression of aquaporin 3 and aquaporin 9 protein was 7.5±2. 0 and 10. 0 ±1.6 in patient group, respectively, while in control group, they were 5.4 ±2.2 and 5.6±2. 1. When compared with those proteins in control group, it exhibited statistical difference (P<0.05). However, in placental trophoblast of patient greup,the expression of aquaporin 3 and aquaporin 9 protein were 3.5±1.4and 4. 0±2. 5, respectively, which were

  14. Expression of aquaporin 8 in human fetal membrane and placenta of idiopathic polyhydramnios%原发性羊水过多产妇胎膜和胎盘组织中水通道蛋白8的表达

    Institute of Scientific and Technical Information of China (English)

    黄锦; 漆洪波


    目的 探讨水通道蛋白8(AQP8)在原发性羊水过多产妇胎膜和胎盘组织中的表达.方法 收集2005年10月-2007年5月重庆医科大学附属第一医院和重庆市妇幼保健院行足月剖宫产分娩(孕37~40周)的原发性羊水过多产妇12例为羊水过多组,同期因社会因素行剖宫产分娩的正常产妇12例为对照组.采用RT-PCR技术检测两组产妇胎膜和胎盘组织中的AQP8 mRNA表达水平;采用免疫组化技术检测两组产妇胎膜和胎盘组织中的AQP8蛋白表达水平.结果 (1)羊水过多组羊膜、绒毛膜和胎盘组织中的AQP8 mRNA表达水平分别为0.78±0.13、0.58±0.10、0.86±0.15,对照组分别为0.39±0.07、0.45±0.09、0.34±0.09,羊水过多组各组织中AQPS mRNA表达水平均高于对照组,两组分别比较,差异均有统计学意义(P0.05).结论 原发性羊水过多产妇羊膜和胎盘组织中AQP8mRNA及蛋白的表达水平均显著高于正常产妇,提示AQP8在产妇羊水量的调节中发挥重要作用.%Objective To determine the expression of Aquaporin 8 (AQP8) in the fetal membrane and placenta of idiopathic polyhydramnios. Methods The amnion, chorion and placenta were collected from 12 term pregnancies with idiopathic polyhydramnios(polyhydramnios group) and 12 term pregnancies who were normal (control group). The expression of AQP8 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). The expression of AQP8 protein was detected by immunohistochemistry. Results The expression of AQP8 mRNA in amnion, chorion and placenta of polyhydramnios group was (0.78±0.13), (0.58±0.10), and (0.86±0.15) respectively, and that of control group was (0.39±0.07 ), (0.45±0.09),and (0.34±0.09) respectively. The expression of AQP8 protein in amnion, chorion and placenta of polyhydramnios group was (0.195±0.024), (0. 170±0.028), and (0. 193±0.024) respectively, and that of control group was (0. 151±0.018), (0.156±0.024), and (0. 152±0

  15. Multicentre epidemiological survey on the incidence of Streptococcus agalactiae in pregnancy

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    Leonardo Lodolo


    Full Text Available ß-hemolytic streptococci group B (GBS cause infections of the urinary tract, endometrium, amnion, and superinfection of wounds, with complications in 2% -4% of cases in pregnant woman.The infection transmitted to newborns is a cause of sepsis and meningitis with high incidence of mortality.A recent report on a multicentric study of the ISS, showed that detection of S. agalactiae in pregnant women in different realities of healthcare, ranging from 3.9% to 19.4%. This survey covers a sample of 29607 women screened in a total of 36852 childbirths, with a positivity of 4183 subjects equal to 14.1%. Thanks to the GBS Study Group of Piemonte and Valle d’Aosta microbiology laboratories, with the support of 42 study centres have been involved. Collection techniques, types of medium, environmental of incubation of the culture, possible enrichment in broth, of the swabs of 28491 women during 2006/2007 have been considered. 24.36% were vaginal swab while 75.64% vagino/rectal. According to data collected at 31 centers with eligible data (22,175 pregnant GBS positivity was 12.7% (CI [Confidence Interval] + / -95%: 13.1% -12.2% with wide variability between individual centers (2% -22.6%. The enrichment broth is used by 6 out of 9 centers with positivity> 15% (M: 20.05%, from 6 to 14 with positivity between 10-15% (M: 12.61%, from none to 8 centers with less than 10% positivity (M: 4.95%.This multicentre survey underlined the preference for vagino/rectal swab, as recommended by the literature. The frequency of GBS positivity of pregnant women on the whole samples is similar to that found in other national surveys, even if with large differences between the different centers.The sensitivity tends to increase with the use of enrichment techniques, whose use is uneven between the centers, perhaps indicating the need for greater uniformity of protocols based on the results of microbiological analysis EBM (Evidence Based Medicine.


    Sirakov, M; Tomova, E


    Mayer-Rokitansky-Küster-Hauser syndrome is a part of the group of abnormalities known as "Anomalies of the fusion of the Müllerian ducts". It is characterized by normal development of breasts and hairs, normal appearance of external genitalia, normal feminine genotype (46XX), normal FSH, LH, E₂ and Test. levels, normal ovaries and congenital absence of uterus and the whole or the upper two thirds of the vagina. It is observed in about 15% of the cases with primary amenorrhea and the incidence is about 1:4500-6000. Etiologycal factors such as lack of estrogen/gestagen receptors, deletions or mutations of genes that stop the fusion, as well as the activation of anti-Müllerian hormone (AMH), are considered. The etiology is being explored but there is no consensus yet. The diagnosis is confirmed during a clinical examination, which takes place because of a primary amenorrhea (often happening at the age of 16), and the absence of a uterus and vagina is proved. The therapy should be handled by a multidisciplinary team including obstetrician, trained midwife, psychologist, specialist in imaging and psycho-sexual counseling. The idea for surgical creation of vagina (neovagina) dates many years ago. The first known documents date back to 1817 and over the years a variety of methods are offered usage of amnion, dura mater, peritoneum, skin grafts, different parts of the intestine, cellulose, etc. The first method of non-surgical treatment is offered by the Czech gynecologist Frank. His ambition was to build a vagina by gradual dilatation of the tissue while applying dilatators with successively increasing length and thickness. The method was further developed by Ingram (1981) and nowadays by Edmonds (2012). He reported about 245 patients treated during the last 12 years by his team. 232 of them had a success in anatomic aspect (95%), 13 did not complete the treatment due to psychological or cultural problems. The experts from American College of Obstetricians and

  17. Under-attending free antenatal care is associated with adverse pregnancy outcomes

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    Heinonen Seppo


    Full Text Available Abstract Background Most pertinent studies of inadequate antenatal care concentrate on the risk profile of women booking late or not booking at all to antenatal care. The objective of this study was to assess the outcome of pregnancies when free and easily accessible antenatal care has been either totally lacking or low in number of visits. Methods This is a hospital register based cohort study of pregnancies treated in Kuopio University Hospital, Finland, in 1989 – 2001. Pregnancy outcomes of women having low numbers (1–5 of antenatal care visits (n = 207 and no antenatal care visits (n = 270 were compared with women having 6–18 antenatal visits (n = 23137. Main outcome measures were: Low birth weight, fetal death, neonatal death. Adverse pregnancy outcomes were controlled for confounding factors (adjusted odds ratios, OR: s in multiple logistic regression models. Results Of the analyzed pregnant population, 1.0% had no antenatal care visits and 0.77% had 1–5 visits. Under- or non-attendance associated with social and health behavioral risk factors: unmarried status, lower educational level, young maternal age, smoking and alcohol use. Chorio-amnionitis or placental abruptions were more common complications of pregnancies of women avoiding antenatal care, and pregnancy outcome was impaired. After logistic regression analyses, controlling for confounding, there were significantly more low birth weight infants in under- and non-attenders (OR:s with 95% CI:s: 9.18 (6.65–12.68 and 5.46 (3.90–7.65, respectively more fetal deaths (OR:s 12.05 (5.95–24.40 and 5.19 (2.04–13.22, respectively and more neonatal deaths (OR:s 10.03 (3.85–26.13 and 8.66 (3.59–20.86, respectively. Conclusion Even when birth takes place in hospital, non- or under-attendance at antenatal care carries a substantially elevated risk of severe adverse pregnancy outcome. Underlying adverse health behavior and possible abuse indicate close surveillance of the

  18. Diagnostic value of ultrasonography for amniotic fluid belt-shaped echo%超声对羊水内带状回声的诊断价值

    Institute of Scientific and Technical Information of China (English)

    马芬; 田荣华; 李燕


    Objective To explore the clinical value of ultrasonography in the diagnosis of amniotic fluid belt-shaped echo. Methods Fifty four mid and late pregnant women with belt-shaped echo in amniotic fluid underwent ultrasonography. Ultrasonographic characteristic of belt-shaped echo in amniotic fluid, the condition of fetuses and placenta appendages were observed. Medical history was inquired and the patients were followed up. Results Among 54 cases with belt-shaped echo in amniotic fluid, 18 of them showed septum for twin pregnancy, 16 of them presented amniotic sheets. The rest included 9 cases of incomplete mediastinum uterus, 7 cases of infused amnion and chorion, 2 cases of lumbar placenta and amniotic band syndrome. Conclusion There are many kinds of reasons for the formation of amniotic fluid belt-shaped echo, some are the normal physiological phenomenon or the benign process, some are the serious pathology phenomena affecting fetal prognosis directly. Correct diagnosis has important value for clinical treatment.%目的:探讨超声诊断羊水内带状回声的临床价值。方法54例伴有羊水内带状回声的中晚期妊娠的孕妇,超声观察带状回声的超声特点及胎儿、附属物情况,询问病史并追踪随访。结果54例羊水内带状回声中,双胎间隔膜18例,羊膜片16例,不全纵隔子宫9例,羊膜、绒毛膜未融合7例,轮状胎盘2例,羊膜带综合征2例。结论羊水内带状回声的形成有多种原因,有的为正常生理现象或良性过程,有的为严重病理现象,直接影响胎儿预后,做出正确诊断对指导临床有重要的价值。

  19. 羊水葡萄糖及白介素-6在胎膜早破中的意义%Determination of Amniotic Fluid Glucose and Interleukin 6in Premature Rupture of Membranes

    Institute of Scientific and Technical Information of China (English)

    陈丽红; 胡继芬; 游彩玲; 马炎辉


    Objective To study the relationship between amniotic cavity infection and the content of glucose and interleukin 6(IL 6) in amniotic fluid in premature rupture of membranes(PMOR) group. Methods To chose randomly 70 examples of PMOR patients without pregnant complication in 1994~1999. Leading by Brightness mode of ultrasound, amniotic fluid was obtained by puncturing the amnion through abdomen. Amniotic fluid glucose was determined quantitatively by "Glucose oxidizing enzyme", IL 6 was measured by enzyme linked immunosorbent assay. Results The content of glucose in amniotic fluid decreased obviously while the quantity of IL 6 increased distinctively in the group with PMOR and amniotic cavity infection. When the content of glucose in amniotic fluid was lower than 0.55 mmol/L while the quantity of IL 6 higher than 315 ng/L, the sensitivity of amniotic cavity infection was expected to be 100%, the specificity over 80%. Conclusion In the PMOR group, to determine the content of glucose and IL 6 in amniotic fluid can predict amniotic cavity infection rapidly and conveniently.%目的探讨胎膜早破患者羊水中葡萄糖及白介素 6(IL 6)含量与宫内感染的关系。方法随机选择1994~1999年70例无妊娠并发症的胎膜早破的患者,在B超引导下,经腹行羊膜腔内穿刺,抽取羊水,用葡萄糖氧化酶方法测定羊水中葡萄糖,用酶联免疫方法测定IL 6含量。结果胎膜早破并宫内感染者,羊水葡萄糖含量明显下降,IL 6含量则明显增高,当羊水葡萄糖含量≤0.55 mmol/L,IL 6含量≥315 ng/L时,预测宫内感染的敏感性100%,特异性>80.00%。结论胎膜早破患者,测定羊水葡萄糖及IL 6含量,能早期快速预测宫内感染。

  20. Study on mechanisms of hypertension in rat adult offspring following prenatal exposure to immuno-inflammatory stimulants

    Institute of Scientific and Technical Information of China (English)

    ZHOU Jian-zhi; LI Xiao-hui


    Objective Essential hypertension (EH) is one of the most common cardiovascular disease and the main causes of human fatility. Recently significant progress has been made in our lab, it was found that exterior stimulation during pregnancy may play a key role in chronicle adult disease. However, what factors affect the growth of fetus after those exterior stimulation and why has not been reported. Based on our previous finding, this study intends to investigate how immuno-inflammatory stimulation affect the development of embryo. Methods 1. Sprague-Dawley (SD) rats, dams in each group received i.p. injections of 0.79 mg· kg-1 LPS, 8 mg·kg-1 zymosan or sterile saline respectively on their gestational days 8, 10, and 12.2. The serums were collected in tail nick at 2 h after the last injection, and the amniotic fluid was mixed at 2, 12, 24,48 h after the last injection. TNF-α and IL-6 levels of serum and amniotic fluid were measured by RIA method. 3. TNF-α and IL-6 mRNA levels were quantitated in amnion, placenta, amniotic fluid, Embryo and maerophage by real-time fluorescent quantitative-PCR. Results 1. The serum level of TNF-α and IL-6 in LPS group and zymosan group was higher than that in control group (P<0.01). It showed that there was immuno-imflammatory response after LPS or zymosan injection in rats. The mRNA levels of TNF-α and IL-6 was very higher in macrophage than in other organization. 2. In embryo, the mRNA level of IL-6 was more than other organization, but the mRNA level of TNF-α was lower than other organization. However, the IL-6 mRNA level of LPS group and zymosan group was higher several dozens times than control group on 24 h and 48 h. Conclusions It suggested that IL-6 was important in the model that prenatalexposure to immuno-inflammatory stimulant results in increases of blood pressure and body weight in rats.

  1. Fetal alcohol exposure and development of the integument

    Directory of Open Access Journals (Sweden)

    Longhurst WD


    Full Text Available William D Longhurst,1 Jordan Ernst,2 Larry Burd3 1Center for Emergency Medicine, University Hospitals Case Medical Center, Case Western Reserve University, Cleveland, OH, USA; 2University of North Dakota School of Medicine and Health Sciences, Grand Forks, ND, USA; 3Department of Pediatrics, North Dakota Fetal Alcohol Syndrome Center, University of North Dakota School of Medicine and Health Sciences, Grand Forks, ND, USA Background: The physiology of fetal alcohol exposure changes across gestation. Early in pregnancy placental, fetal, and amniotic fluid concentrations of alcohol exposure are equivalent. Beginning in mid-pregnancy, the maturing fetal epidermis adds keratins which decrease permeability resulting in development of a barrier between fetal circulation and the amniotic fluid. Barrier function development is essential for viability in late pregnancy and in the extra-uterine environment. In this paper we provide a selected review of the effects of barrier function on fetal alcohol exposure. Methods: We utilized a search of PubMed and Google for all years in all languages for MeSH on Demand terms: alcohol drinking, amnion, amniotic fluid, epidermis, ethanol, female, fetal development, fetus, humans, keratins, permeability, and pregnancy. We also reviewed the reference lists of relevant papers and hand-searched reference lists of textbooks for additional references. Results: By 30 gestational weeks, development of barrier function alters the pathophysiology of ethanol dispersion between the fetus and amniotic fluid. Firstly, increases in the effectiveness of barrier function decreases the rate of diffusion of alcohol from fetal circulation across fetal skin into the amniotic fluid. This reduces the volume of alcohol entering the amniotic fluid. Secondly, barrier function increases the duration of fetal exposure by decreasing the rate of alcohol diffusion from amniotic fluid back into fetal circulation. Ethanol is then transported into

  2. Genetic and biochemical evidence that gastrulation defects in Pofut2 mutants result from defects in ADAMTS9 secretion. (United States)

    Benz, Brian A; Nandadasa, Sumeda; Takeuchi, Megumi; Grady, Richard C; Takeuchi, Hideyuki; LoPilato, Rachel K; Kakuda, Shinako; Somerville, Robert P T; Apte, Suneel S; Haltiwanger, Robert S; Holdener, Bernadette C


    Protein O-fucosyltransferase 2 (POFUT2) adds O-linked fucose to Thrombospondin Type 1 Repeats (TSR) in 49 potential target proteins. Nearly half the POFUT2 targets belong to the A Disintegrin and Metalloprotease with ThromboSpondin type-1 motifs (ADAMTS) or ADAMTS-like family of proteins. Both the mouse Pofut2 RST434 gene trap allele and the Adamts9 knockout were reported to result in early embryonic lethality, suggesting that defects in Pofut2 mutant embryos could result from loss of O-fucosylation on ADAMTS9. To address this question, we compared the Pofut2 and Adamts9 knockout phenotypes and used Cre-mediated deletion of Pofut2 and Adamts9 to dissect the tissue-specific role of O-fucosylated ADAMTS9 during gastrulation. Disruption of Pofut2 using the knockout (LoxP) or gene trap (RST434) allele, as well as deletion of Adamts9, resulted in disorganized epithelia (epiblast, extraembryonic ectoderm, and visceral endoderm) and blocked mesoderm formation during gastrulation. The similarity between Pofut2 and Adamts9 mutants suggested that disruption of ADAMTS9 function could be responsible for the gastrulation defects observed in Pofut2 mutants. Consistent with this prediction, CRISPR/Cas9 knockout of POFUT2 in HEK293T cells blocked secretion of ADAMTS9. We determined that Adamts9 was dynamically expressed during mouse gastrulation by trophoblast giant cells, parietal endoderm, the most proximal visceral endoderm adjacent to the ectoplacental cone, extraembryonic mesoderm, and anterior primitive streak. Conditional deletion of either Pofut2 or Adamts9 in the epiblast rescues the gastrulation defects, and identified a new role for O-fucosylated ADAMTS9 during morphogenesis of the amnion and axial mesendoderm. Combined, these results suggested that loss of ADAMTS9 function in the extra embryonic tissue is responsible for gastrulation defects in the Pofut2 knockout. We hypothesize that loss of ADAMTS9 function in the most proximal visceral endoderm leads to slippage of

  3. Effect of arginine and threonine administered in ovo on digestive organ developments and subsequent growth performance of broiler chickens. (United States)

    Tahmasebi, S; Toghyani, M


    This trial was conducted to investigate the effect of arginine (Arg), threonine (Thr) and Arg + Thr administered in ovo on growth performance, digestive organs and intestinal morphology in broiler chickens. On day 14 of incubation, 400 fertile eggs were randomly allotted to five experimental treatments through injection in the amniotic fluid including: (i) control (none injected), (ii) sham (0.5 ml of 0.5% saline), (iii) Arg (35 mg/egg), (iv) Thr (25 mg/egg) and (v) Arg + Thr (35 + 25 mg/egg). After hatching, chicks were given a commercial corn-soya bean diet up to 42 days of age. Daily feed intake (FI) and body weight (BW) of chicks were measured during different periods of the trial. Digestive organs were measured for their relative weight and intestinal length on days 11 and 42 of age. Intestinal morphometric traits were evaluated on day 11 of the experiment. Supplementing amino acids affected the performance of broiler chicks as Thr significantly increased FI and BW across starter, grower and finisher periods compared with sham and control (p  0.05); however, amino acid supplemented groups had lower villus height than control in jejunum (p pancreas (r = -0.484; p = 0.030), duodenal (r = -0.577; p = 0.007) as well as ileal lengths (r = -0.471; p = 0.035) and FI of entire period. Otherwise, positive relationships were observed between duodenum (r = 0.580; p = 0.007) and ileum (r = 0.582; p = 0.007) weights on day 42 and FI of chickens across the entire phase. In conclusion, Arg and particularly Thr injection into amnion can improve FI and post-hatch growth performance of chickens which may be mediated by the development of digestive organs. PMID:26608576

  4. 血清学筛查与胎儿超声检查在18、13三体综合征产前诊断中的临床应用%Clinical applications of serological screening and fetal ultrasonography for prenatal diagnosis of trisomy 18 and trisomy 13

    Institute of Scientific and Technical Information of China (English)

    梁学清; 韦丽萍; 唐娟


    Objective: To evaluate the values of serological screening and fetal ultrasonography in prenatal diagnosis of trisomy 18 and trisomy 13. Methods:A total of 780 pregnant women received serological screening and fetal ultrasonography, the samples of amnion fluid were obtained by amniocentesis, the cell culture and chromosomal karyotype analysis were conducted for prenatal diagnosis. Results; Among 780 fetuses, 6 fetuses were found with trisomy 18 and trisomy 13, the incidence was 0.77% , including 3 fetuses with trisomy 18 and 3 fetuses with trisomy 13. Three fetuses with trisomy 18 were found with high risk of serological screening and abnormal ultrasonic structure; and the other three fetuses were found with abnormal ultrasonic structure and low risk of serological screening. Conclusion: Serological screening of pregnant women combined with fetal ultrasonography is an effective method to detect trisomy 18 and trisomy 13 before delivery.%目的:评价利用孕妇血清学筛查与胎儿超声检查在18、13三体综合征胎儿产前诊断的价值.方法:对780例孕妇进行孕妇血清学筛查与胎儿超声检查,羊膜腔穿刺取羊水进行细胞培养染色体核型分析进行产前诊断.结果:780例胎儿中共发现6例18、13三体综合征,发生率为0.77%.其中3例18三体综合征,3例13三体综合征.3例18三体综合征血清学筛查高风险和超声结构异常,其余3例超声检查发现结构异常但血清学筛查为低风险.结论:孕妇血清筛查结合胎儿超声检查是产前检出18、13三体综合征胎儿的有效检查方法.

  5. Liposome-mediated IL-28 and IL-29 expression in A549 cells and antiviral effect of IL-28 and IL-29 on WISH cells

    Institute of Scientific and Technical Information of China (English)

    Ming-cai LI; Hao-yang WANG; Hai-yan WANG; Tao LI; Shao-heng HE


    Aim:To construct the recombinant vectors carrying interleukin (IL) -28A,IL-28B and IL-29 cDNAs and express them in human A549 cells,and analyze their antiviral activity in vesicular stomatitis virus (VSV)-infected human immortalized amnion epithelial cell line (WISH cells).Methods:Total cell RNA was extracted from human peripheral blood mononuclear cells (PBMC) activated with poly I:C.The cDNAs encoding human IL-28A.IL-28B.and IL-29 were amplified by reversetranscription polymerase chain reaction (RT-PCR) and inserted into pcDNA3.1/V5-His-TOPO vectors.These recombinant vectors were transfected into human A549 cells by a liposome-mediated gene transfer method.Semiquantitative RT-PCR and Westem blotting were used to detect the mRNA and protein expression of IL-28A,IL-28B,and IL-29.The antiviral activity of IL-28A,IL.28B,and IL-29 was determined by a cytopathic eflfect reduction assay on WISH cells using VSV as a challenge virus.Results:The DNA sequences of the inserts were identical to the published sequences encoding IL-28A,IL-28B,and IL-29 in GenBank.It was transfected cells.Expression of all 3 interleukins in A549 cells was confirmed by Wlestem blot analysis.IL-28 and IL-29 expressed by A549 cells.1ike interferon (IFN)α-2b,were able to protect WISH cells against VSV infection.Conclusion:IL-28 and IL-29 cDNAs were successfully cloned and expressed in eukaryotic cells via transfection with pcDNA3.1/V5-His-TOPO-IL-28/IL-29.Transfection with this vector produced a specific antiviral activity similar to that of IFN-α.which will provide a new tool for the functional study of these cytokines in humans.

  6. Clinical analysis of occupational corneal burns%职业性角膜灼伤临床分析

    Institute of Scientific and Technical Information of China (English)

    杨水平; 曹国平; 窦裕平


    Objective To study the characteristics and treatment of occupational corneal burns.Methods 85 eyes of 75 patients with corneal burns were analyzed retrospectively.The treatment included topical eye drops,single layer amnion covering or amniotic tamponade according to the scope and depth of corneal injury.All the patients were followed up for 6 months after treatment.Results Corneal healing and visual recovery after injury had close relationship with corneal burns depth:the cornea and vision recovered well when burns involved superficial cornea only; Corneal healing and vision recovery were bad when burns involved deep cornea.In which corneal melt occurred and enucleation of eyeball were perfermed in some cases even after active operation had done.Conclusion Occupational corneal burns in factory is severe and the efficacy of treatment is poor; Prevention is important and it is necessary to supervise and manage all production links.%目的 探讨职业性角膜灼伤特点及治疗效果.方法 回顾性分析我院附近工厂因角膜灼伤就诊我科门诊及住院75例(85眼),根据角膜损害范围及深度,采取局部滴眼液、单层羊膜覆盖或羊膜填塞覆盖等不同治疗措施,治疗后随访6月.结果 伤后角膜愈合及视力恢复与角膜灼伤累及的深度关系密切:灼伤仅累及角膜上皮及基质浅层者,角膜及视力恢复良好;灼伤累及角膜基质深层,即使积极进行手术,术后部分患者仍出现角膜溶解,甚至眼球摘除,角膜愈合及视力恢复不理想.结论 工厂的角膜灼伤伤情严重,总体治疗效果欠佳;重在预防,有必要对生产各个环节进行监督及管理.

  7. 水通道蛋白9与羊水量异常的研究进展%Research progress on relationship between expression of aquaporin 9 and amniotic fluid volume abnormality

    Institute of Scientific and Technical Information of China (English)

    解雁飞; 李红梅


    水通道蛋白普遍存在于人体组织器官的细胞膜上,跨过脂质双分子层转运水及其它小分子物质,调节人体内的水平衡代谢。近年来研究发现水通道蛋白是羊水平衡的重要通道,参与母胎的液体交换。水通道蛋白9作为水通道蛋白家族中重要成员之一,目前被认为表达于人类的胎膜和胎盘,可能是羊水膜内调节中的一种重要的水通道。对水通道蛋白9在人胎盘胎膜的分布以及水通道蛋白9与羊水量异常相关性的深入研究,有助于探索羊水量异常的发生机制,从而为治疗羊水量异常提供新的思路和方向。为进一步明确羊水量异常的发病机制以指导临床的诊治,该文就国内外水通道蛋白9及其与羊水量异常关系的研究进展进行了综述。%Aquaporin ( AQP ) , which exists widely in cell membrane of human tissues and organs , passes through lipidic bilayer ( phospholipid bilayer ) to transport water and other small molecules and regulates water metabolic balance in body .Recent studies have found that AQP is an important passage for amnion fluid balance and maternal-fetal fluid exchange .As one of the important members of the AQPs family, AQP9 is now believed to be expressed in human fetal membrane and placenta , and it may be an important water passage for amniotic fluid intramembrane regulation .Deep investigationes on distribution of AQP 9 in fetal membranes and placenta and its relationship with amniotic fluid volume abnormality is contributive to explore mechanisms of amniotic fluid volume abnormality , so as to provide a new idea and direction for the treatment .In order to further clarify the pathogenesis of abnormal amniotic fluid volume and to guide the clinical diagnosis and treatment , the article reviewed recent advance both at home and abroad in AQP 9 and its relationship with amniotic fluid volume abnormality .

  8. Clinical exploration of the orbital Implant at its early rtage after retinoblastoma operation%视网膜母细胞瘤术后羟基磷灰石义眼座一期植入探讨

    Institute of Scientific and Technical Information of China (English)



    目的:探讨儿童视网膜母细胞瘤(retinoblastoma,RB)眼球摘除术后羟基磷灰石义眼座一期眶内植入的可行性.方法:对21例(21眼)RB患儿,施行眼球摘除联合无包裹羟基磷灰石义眼座一期眶内植入手术.结果:随访3个月至5年,义眼座活动度良好,外形满意,眼眶及面部发育正常.2例眼座暴露,其中1例暴露4 mm×5 mm,应用重组人表皮细胞生长因子滴眼液治疗,1例暴露9 mm ×9 mm,给予手术介入+羊膜覆盖,均治愈.结论:RB术后羟基磷灰石义眼座一期植入效果好,有助于儿童眼眶及面部发育,不影响放射治疗及CT检查,最常见并发症为眼座暴露.%Objective:To explore the possibility of the hydroxyapatite orbital implants after children-retinoblastoma-eyeball-excision operations. Methods: For 21 cases ( 21 eyes ) with retinoblastoma, enucleation was executed with the exposed hydroxyapatite orbital implant at its early stage. Results:Three-month to five-year subsequent visits showed that the orbitals acted well in good shape and developed desirably in the orbit and the face. For the two cases with orbitals exposure. One exposed 4 mm × 5 mm, cured with the treatment of restoratin-superficial-skin-gowth-factor eyeliquid, the other exposed 9 mm × 9 mm, cured with the operation of amnion cover. Conclusions:The hydroxyapatite orbital implants had satisfactory effects after retinoblastma operations, with good help for childrren's orbits and faces growth,with little influence on radiotherapy or CT examination. The most common complication is the eye exposure.

  9. Novel concepts on pregnancy clocks and alarms: redundancy and synergy in human parturition. (United States)

    Menon, Ramkumar; Bonney, Elizabeth A; Condon, Jennifer; Mesiano, Sam; Taylor, Robert N


    The signals and mechanisms that synchronize the timing of human parturition remain a mystery and a better understanding of these processes is essential to avert adverse pregnancy outcomes. Although our insights into human labor initiation have been informed by studies in animal models, the timing of parturition relative to fetal maturation varies among viviparous species, indicative of phylogenetically different clocks and alarms; but what is clear is that important common pathways must converge to control the birth process. For example, in all species, parturition involves the transition of the myometrium from a relaxed to a highly excitable state, where the muscle rhythmically and forcefully contracts, softening the cervical extracellular matrix to allow distensibility and dilatation and thus a shearing of the fetal membranes to facilitate their rupture. We review a number of theories promulgated to explain how a variety of different timing mechanisms, including fetal membrane cell senescence, circadian endocrine clocks, and inflammatory and mechanical factors, are coordinated as initiators and effectors of parturition. Many of these factors have been independently described with a focus on specific tissue compartments.In this review, we put forth the core hypothesis that fetal membrane (amnion and chorion) senescence is the initiator of a coordinated, redundant signal cascade leading to parturition. Whether modified by oxidative stress or other factors, this process constitutes a counting device, i.e. a clock, that measures maturation of the fetal organ systems and the production of hormones and other soluble mediators (including alarmins) and that promotes inflammation and orchestrates an immune cascade to propagate signals across different uterine compartments. This mechanism in turn sensitizes decidual responsiveness and eventually promotes functional progesterone withdrawal in the myometrium, leading to increased myometrial cell contraction and the

  10. Transplantation of human amniotic epithelial cells improves hindlimb function in rats with spinal cord injury

    Institute of Scientific and Technical Information of China (English)

    WU Zhi-yuan; HUI Guo-zhen; LU Yi; WU Xin; GUO Li-he


    Background Human amniotic epithelial cells (HAECs), which have several characteristics similar to stem cells,therefore could possibly be used in cell therapy without creating legal or ethical problems. In this study, we transplanted HEACs into the injured spinal cord of rats to investigate if the cells can improve the rats' hindlimb motor function.Methods HAECs were obtained from a piece of fresh amnion, labeled with Hoechst33342, and transplanted into the site of complete midthoracic spinal transections in adult rats. The rats (n=21) were randomly divided into three groups: Sham-operation group (n=7), cells-graft group (n=7), and PBS group (n=7). One rat of each group was killed for histological analysis at the second week after the transplantation. The other six rats of each group were killed for histological analysis after an 8-week behavioral testing. Hindlimb motor function was assessed by using the open-field BBB scoring system. Survival rate of the graft cells was observed at second and eighth weeks after the transplantation. We also detected the myelin sheath fibers around the lesions and the size of the axotomized red nucleus. A one-way ANOVA was used to compare the means among the groups. The significance level was set at P<0.05.Results The graft HAECs survived for a long time (8 weeks) and integrated into the host spinal cord without immune rejection. Compared with the control group, HAECs can promote the regeneration and sprouting of the axons, improve the hindlimb motor function of the rats (BBB score: cells-graft group 9.0± 0.89 vs PBS group 3.7± 1.03, P<0.01), and inhibit the atrophy of axotomized red nucleus [cells-graft group (526.47 ± 148.42) μm2 vs PBS group (473.69±164.73) μm2, P<0.01].Conclusion Transplantation of HAECs can improve the hindlimb motor function of rats with spinal cord injury.

  11. Feasibility and clinical significance of lamellar body count in amniotic fluid for prediction of fetal lung maturity%羊水板层小体计数预测胎儿肺成熟的可行性及其临床意义

    Institute of Scientific and Technical Information of China (English)

    张温麂; 温岩; 孙晓峰


    Objective: To explore the clinical application value of lamellar body count in amniotic fluid for prediction of fetal lung maturity. Methods: A total of 156 single pregnant women were selected as study objects, the amniotic fluid samples underwent LBC count, L/S detection, and foam test simultaneously during 28-38 gestational weeks. Results: There was a positive linear correlation between LBC count and L/S ratio (γ = 0.623, P 0.05); the predictive values of LBC count was significantly higher than those of foam test ( P < 0. 01) . Conclusion: Amnionic fluid LBC count is superior to routine foam test for prediction of fetal lung maturity; compared with L/S ratio, the method is time - saving and labor - saving, and it is the first choice to predict fetal lung maturity in clinic.%目的:探讨羊水板层小体计数预测胎肺成熟度的临床应用价值.方法:选择156例单胎孕妇作为研究对象,孕28 - 38周,每份羊水分3份分别进行LBC计数、L/S比值测定及泡沫试验.结果:羊水LBC与L/S比值成正线性相关,相关系数(r)为0.623,有极显著统计学意义(P<0.001);LBC与泡沫试验呈等级相关,相关系数为(r)为0.781,有极显著统计学意义(P<0.001).以LBC≤10×109/L作为判断胎儿肺不成熟的临界值,以LBC≥35×109/L作为判断胎儿肺成熟的临界值,比较羊水LBC计数各预测值率与L/S比值差异无统计学意义(P>0.05);与泡沫试验相比,羊水LBC计数的各预测值明显高于泡沫试验,差异有统计学意义(P<0.01).结论:羊水LBC计数在预测胎肺成熟度方面优于传统的泡沫试验,较L/S比值省时、省力,是临床中用于判断胎儿肺成熟方法的首选.

  12. The clinical significance of ultrasonography in the diagnosis of simple polyhydramnios%超声诊断单纯性羊水过多的临床意义

    Institute of Scientific and Technical Information of China (English)

    谭莉娜; 肖春梅; 冉素真; 林芸


    目的:探讨超声诊断单纯性羊水过多的临床意义。方法以二维半定量法将该院住院及门诊首次超声诊断单纯性羊水过多的186例单胎孕妇分为3组:单独羊水指数(AFI)≥20 cm(A组)、单独最大羊水池深度(AFV)≥8 cm(B组)、AFI≥20 cm且AFV≥8 cm(C组),并动态监测其变化。结果 C组中持续性羊水过多较其他两组发生率高,胎儿畸形发生率亦较其他两组发生率高,差异均有统计学意义(P<0.05);3组中未发现复发性羊水过多与胎儿畸形有相关性(P>0.05);84.4%的单纯性羊水过多未发现胎儿畸形。结论超声诊断单纯性羊水过多对评价胎儿畸形、宫内转归及预后具有一定的临床价值,利于产前咨询及处理。%Objective To investigate the clinical significance of ultrasonography in the diagnosis of simple polyhydramnios . Methods 186 singleton pregnancies cases of inpatient and outpatient diagnosed with simple polyhydramnios were divided into three groups with a two-dimensional semi-quantitative method ,separate amniotic fluid index (AFI)> 20cm (A group) ,separate maxi-mum amnionic fluid volume(AFV)>8 cm(B group) ,AFI≥20 cm and AFV≥8 cm(C group) ,and the changes were dynamically monitored .Results Occurrence rates of persistent polyhydramnios and fetal malformations in C group was higher than in the other two groups ,and the difference showed no statistical significance(P0 .05);Among the 84 .4% of simple polyhydramnios ,no fetal malfor-mation was found .Conclusion Ultrasound diagnosis of simple polyhydramnios shows clinical value in assessment of fetal malfor-mation ,fetal outcome ,perinatal morbidity and prognosis ,and is good for prenatal counseling and treatment .

  13. 长沙市2001-2010年孕产妇死亡分析%Analysis of Maternal Mortality in Changsha City from 2001 to 2010

    Institute of Scientific and Technical Information of China (English)

    祖月娥; 谢日学; 李爱林; 周红女; 徐扬; 林蓓蓓


    obstetric hemorrhage, pregnancy with internal and surgical diseases, hypertensive disorder complicating pregnancy, and amnionic fluid embolism. As the first factor affecting maternal death, obstetric hemorrhage had superseded pregnancy with internal and surgical diseases. Conclusions The effective countermeasures to reduce the maternal mortality are as follows: strengthening maternal care and management, perfecting the maternal and child health network, raising hospital delivery rate, enhancing the management of high-risk pregnancy, and improving the maternal - emergency ability of township and county health care centers.

  14. Membrana amniótica preservada em glicerina no reparo de feridas cutâneas de membros locomotores de eqüinos Equine amniotic membrane preserved in glycerin in reparing the equine limbs cütaneous wounds

    Directory of Open Access Journals (Sweden)

    Valdemir Alves de Oliveira


    treated with amniotic membrane, and the other, the control group, treated only with a damp gauze changed every 48 hours. In all the wounds the procedure was to evaluate the measurement of the área, degree of secretion, development of exuberant granulation tissue and the complete time of epitheliazation. Bacteriologic and histopatologic exams were made in a sample of the treated and contrai groups. The biochemical determination of proteins and the identification of fibroblastic growth factor were realized in the amnionic membrane. In all the treated wounds were observed shorter healing time, less formation of exuberant granulation tissue, smaller incidence of pathogenic bacterias and a smaller increase of the wound area.

  15. Zinc ferrite nanoparticles activate IL-1b, NFKB1, CCL21 and NOS2 signaling to induce mitochondrial dependent intrinsic apoptotic pathway in WISH cells

    Energy Technology Data Exchange (ETDEWEB)

    Saquib, Quaiser; Al-Khedhairy, Abdulaziz A.; Ahmad, Javed; Siddiqui, Maqsood A.; Dwivedi, Sourabh; Khan, Shams T. [Department of Zoology, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451 (Saudi Arabia); Chair for DNA Research, Department of Zoology, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451 (Saudi Arabia); Musarrat, Javed, E-mail: [Chair for DNA Research, Department of Zoology, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451 (Saudi Arabia); Department of Agricultural Microbiology, Faculty of Agricultural Sciences, Aligarh Muslim University, Aligarh 202002, U.P. (India)


    The present study has demonstrated the translocation of zinc ferrite nanoparticles (ZnFe{sub 2}O{sub 4}-NPs) into the cytoplasm of human amnion epithelial (WISH) cells, and the ensuing cytotoxicity and genetic damage. The results suggested that in situ NPs induced oxidative stress, alterations in cellular membrane and DNA strand breaks. The [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) and neutral red uptake (NRU) cytotoxicity assays indicated 64.48 ± 1.6% and 50.73 ± 2.1% reduction in cell viability with 100 μg/ml of ZnFe{sub 2}O{sub 4}-NPs exposure. The treated WISH cells exhibited 1.2-fold higher ROS level with 0.9-fold decline in membrane potential (ΔΨm) and 7.4-fold higher DNA damage after 48 h of ZnFe{sub 2}O{sub 4}-NPs treatment. Real-time PCR (qPCR) analysis of p53, CASP 3 (caspase-3), and bax genes revealed 5.3, 1.6, and 14.9-fold upregulation, and 0.18-fold down regulation of bcl 2 gene vis-à-vis untreated control. RT{sup 2} Profiler™ PCR array data elucidated differential up-regulation of mRNA transcripts of IL-1b, NFKB1, NOS2 and CCL21 genes in the range of 1.5 to 3.7-folds. The flow cytometry based cell cycle analysis suggested the transfer of 15.2 ± 2.1% (p < 0.01) population of ZnFe{sub 2}O{sub 4}-NPs (100 μg/ml) treated cells into apoptotic phase through intrinsic pathway. Over all, the data revealed the potential of ZnFe{sub 2}O{sub 4}-NPs to induce cellular and genetic toxicity in cells of placental origin. Thus, the significant ROS production, reduction in ΔΨm, DNA damage, and activation of genes linked to inflammation, oxidative stress, proliferation, DNA damage and repair could serve as the predictive toxicity and stress markers for ecotoxicological assessment of ZnFe{sub 2}O{sub 4}-NPs induced cellular and genetic damage. - Highlights: • First report on the molecular toxicity of ZnFe{sub 2}O{sub 4}-NPs in cells of placental origin • WISH cells treated with ZnFe{sub 2}O{sub 4}-NPs exhibited cytoplasmic

  16. Analysis of risk factors for brain injury in preterm infants after preterm premature rupture of membranes%未足月胎膜早破后早产儿脑损伤的相关危险因素分析

    Institute of Scientific and Technical Information of China (English)

    卢红艳; 常明; 吴丽华


    目的:分析未足月胎膜早破后早产儿脑损伤的高危因素.方法:对2008年1月~ 2010年10月在医院产科出生且生后即转入新生儿科的未足月胎膜早破后早产儿及母亲的临床资料进行回顾性分析,按早产儿生后有无脑损伤分为脑损伤组与无脑损伤组.对患儿胎龄、出生体质量、性别、母亲有无绒毛膜羊膜炎、羊水指数、母亲产前应用激素、破膜时间、分娩方式、胎儿宫内窘迫、窒息复苏10个项目进行分析.结果:137例早产儿符合研究标准,未足月胎膜早破后早产儿脑损伤发生率24.8%.早产儿脑损伤危险因素Logistic回归分析按OR值排序,依次为胎龄、出生体重、窒息复苏及母亲绒毛膜羊膜炎.破膜时间长短与脑损伤无关.结论:未足月胎膜早破后早产儿脑损伤的发生与多因素有关,胎龄越小、体重越低脑损伤发生率越高,母亲绒毛膜羊膜炎及患儿生后有窒息复苏史也为早产儿脑损伤高危因素.%Objective; To explore the high risk factors of brain injury in preterm infants after preterm premature rupture of membranes. Methods: Data were collected retrospectively among preterm infants after preterm premature rupture of membranes who were borne in obstetrics and admitted to neonatal intensive care unit in our hospital from Jan. 2008 to Oct. 2010. All subjects were divided into brain injury group and no brain injury group. Logistic regression was adopted to analyze 10 factors; gestational age, birth weigh, sex, mother chorio-amnionitis, amniotic fluid index, antenatal steroids therapy to mothers, latency after preterm premature rupture of membranes, delivery ways, fetal distress in uterus, asphyxiate resuscitation. Results: Among 137 preterm infants, 33 cases ( 24.8% ) were detected with brain injury. Risk factors for brain injury in preterm infants after preterm premature rupture of membranes were arranged as follows according to odds ratio (OR) value

  17. Clinical analysis 0f 83 cases plant ocular trauma cause fungal keratitis%83例植物性角膜外伤致真菌性角膜炎的临床探讨

    Institute of Scientific and Technical Information of China (English)



    Objectiveo To investigate the means and clinical efficacy of cornea plant trauma cause fungal keratitis. Methods Cornea for 83 cases (83 eyes) plant trauma , cause infectivity keratitis cases reported , Make use of directly microscopy or through a form of the corneal ulcer or through epiphyte culture in laboratory, the choice of treatment.83 cases among the amnion transplantation combined clean out ulcer 22 cases, Tyro transplantation of corneal 8 cases, All patients iodized to cauterize combined clean out ulcer. Partial and full body conventional use to resist fungi drug treatment. Results 83 cases of 81 cases of cure , To cure rate 97.5%, Eyesight are increased .2 cases of reatment when corneal has extensive damage, To close to punch. The front room is thick with pus, Finally illness got worse and had to choose their eyeballs removal. Conclusions In Fungal keratitis reasonable drug treatment , applications , the innovation of surgical techniques and surgical methods the correct choice , is the key to effective infection control and prevent recurrence of infection.%目的探讨植物性角膜外伤致真菌性角膜炎的治疗方法和效果.方法对83例(83只眼)有角膜植物伤史的感染性角膜炎病例,通过直接镜检或根据角膜溃疡的表现或通过实验室真菌培养等方式予以确诊后,选择治疗方案.83例中行溃疡面清除联合羊膜移植术的22例,行板层角膜移植术的8只眼,所有病人行溃疡面清除加碘酊烧灼,局部和全身常规使用抗真菌药物治疗.结果83例病例中81例治愈,治愈率97.5%,视力均较前增加.2例就诊时角膜已大面积毁坏,接近穿孔,前房大量积脓,最终病情恶化而不得不选择眼球内容摘除术.结论在真菌性角膜炎的治疗中,药物的合理应用,手术技巧的革新和手术方式的正确选择,是有效控制感染并防止感染复发的关键.

  18. 羊膜带综合征相关畸形的相关因素、诊断及处理结果分析%Amniotic band syndrome deformity related factors, diagnosis, and treatment results

    Institute of Scientific and Technical Information of China (English)

    梁莉; 黄桂芳; 黄贞; 陆启升


    , abdominal wall defect edge for amniotic membrane gradually to cutaneous squamous epithelial migration ( amniotic - ectoderm transition zone). Another example is a simple meningoencephalocele malformations, all cases of B ultrasound imaging in amniotic fluid can be seen floating band echo, the part attached to the fetus; amniotic band adhesions of fetal limb portion of visible deformity deformity, often to scan the amniotic band echo & the adhesion. Conclusion: ABS mainly for outer surface of tissue and organ defects; involvement of tissue and organ because of its occurrence in the critical period, because of the amniotic sac and amniotic membrane rupture with adhesions interference and teratogenicity j the severity of the deformity and range depends on the amnion rupture time and site. ABS prevention, pregnancy trimester routine ultrasound screening, early diagnosis, early intervention.

  19. Safflower injection-preserved human amniotic membrane is applied in skin wound%红花注射液保存人胎羊膜贴敷皮肤切口*★

    Institute of Scientific and Technical Information of China (English)

    罗秋燕; 李慧智; 边文玲; 王莉莉


    after wound (P < 0.05). The infection rate of skin wound in the rats treated with human amnion saved with safflower injection (5%) was less than that of the other groups at day 5 after wound (P < 0.05). Results showed that the human amniotic membrane preserved by safflower injection could not only enhance the expression of vascular endothelial growth factor around the area of injury in the initial stage, but also reduce the infection rate of skin wound, thereby promoting wound healing in rats.

  20. Optimization of In Vitro Culture Conditions for Human Amniotic Epithelial Cells and Expression of Stem Cell Markers%羊膜上皮细胞体外培养条件的优化及其干细胞标志的表达

    Institute of Scientific and Technical Information of China (English)

    陈宥艺; 陆琰; 王科; 王琰; 吴东颖; 刘兵; 杨瑛; 吕双红


    本研究优化人足月胎盘羊膜上皮细胞(human amniotic epithelium cells,hAEC)的体外培养方法并现察hAEC的干细胞标志的表达情况.取健康产妇足月剖宫产术后的羊膜,采用胰酶多次消化获取hAEC,分别应用10% FBS的DMEM、类似胚胎干细胞的培养条件及添加表皮细胞生长因子(epidermal growth factor,EGF)的类似胚胎干细胞的培养条件对其进行原代和传代培养,观察培养后细胞形态,并通过流式细胞术检测、细胞免疫荧光染色等方法对培养细胞的干细胞多能性标志进行鉴定.结果表明:在类似胚胎干细胞培养条件的基础上添加10 ng/ml EGF可提高hAEC原代细胞的贴壁率和增殖能力,与广泛应用的10% FBS的DMEM培养条件相比,在此培养条件下细胞传代能力显著增强,传至5代仍保持上皮细胞表型,细胞形态在传代过程中改变不明显,并且表达胚胎干细胞全能性的表面标志SSEA-4同时细胞免疫荧光染色显示培养后的hAEC表达波形蛋白(vimentin).结论:采用类似胚胎干细胞的培养条件有利于hAEC在体外的扩增和传代,EGF对其增殖和传代有促进作用,培养的hAEC表达胚胎干细胞多能性标志.%This study was purposed to optimize the culture conditions of the human amniotic epithelium cells (hAEC) in vitro, and detect the expression of hAEC pluripotent markers.Amnion tissues were separated from the underlying chorion through the spongy layer immediately after elective cesarean section of healthy pregnancy women at term.After the subsequent exposure to trypsin digestion, hAEC were cultured in DMEM with different supplements.The growth and proliferation potential of hAEC was evaluated, and the expression of cultured hAEC pluripotent markers was detected by using flow cytometry and immunohistochemistry methods.The results indicated that when being cultured in the mediums similar to that of embryonic stem cell culture supplemented with 10 ng/ml EGF, the h

  1. 体位引流在清理新生儿呼吸道中的作用%Roles of Postural Drainage in the Clearing of Respiratory Tract of Newborns

    Institute of Scientific and Technical Information of China (English)

    吴慧; 徐小珍


    Objective To discuss and analyze the use of postural drainage in the clearing of respiratory tract of newhorns. Methods 600 cases of mature newborns during June to December 2010 from vaginal delivery in Yixing People's Hospital were selected and distributed into research group or control group randomly,300 cases in each group. The research group were put in the foot-high horizontal position to carry out the postural drainage and manual press and massage for mouth and nose to clear the respiratory tract, while mucous extractor plus manual press and massage for mouth and nose were used for the control group. Results The clearing time for the research group was obviously shorter than for the control group. Mostly,the clearing of the respiratory tract can be done at one time to avoid the use of mucous extractor. The rate of asphyxia of newborns of the research group was significantly lower than the rate of the control group. The newborns of the research group suck earlier than the ones of the control group. Moreover, there was no case of amnionic fluid aspiration syndrome in the research group,with statistical significance( all P <0. 05 ). Conclusion Manural press and massage for mouth and nose combined with the foot-high horizontal position may benefit from gravity action and be more effective in clearing the respiratory tract of newborns, markedly reducing the anoxic damage fromor using mucous extractor, the amniotic fluid aspiration and the rate of asphyxia of newborns.%目的 探讨体位引流在刚分娩新生儿清理呼吸道中的作用.方法 选择2010年6 ~12月在宜兴市人民医院阴道分娩的足月新生儿600例.随机分为研究组和对照组,每组300例.研究组使用头低足高位体位引流的基础上配合手法挤压口鼻的方法清理新生儿呼吸道,对照组使用手法挤压加吸痰管吸引的方法.结果 研究组清理呼吸道时间明显短于对照组;且大部分能一次性清理干净呼吸道,而无需再使用

  2. Estudo por microscopia eletrônica do epitélio de neovaginas confeccionadas com membrana amniótica e pesquisa de receptores para estrogênios Electron microscope study of the epithelium of neovagina constructed with amniotic membrane: estrogen receptor evaluation

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    Mauri José Piazza


    cortes histológicos obtidos em biópsias de neovaginas revestidas com membrana amniótica.Purpose: to verify changes in amnion epithelia after neovaginoplasty and the presence of estrogen receptor in this tissue using electron microscopy (TEM. Methods: a group consisting of 33 young patients with vaginal agenesy, most of them amenorreic, either incapable or having difficulty to adequately perform sexual activity, underwent surgical correction of the anomaly by the McIndoe-Banister technique. In each woman, a cavity was created through an opening of the vesicorectal space. A mold made of artificial sponge with a condom recovered by amniotic membrane was introduced into the cavity. After eight days, the mold was removed, leaving a developing neoepithelium from the amniotic membrane. Results: The TEM conducted in samples of tissue obtained from neovaginas showed similar results in all three layers of these epithelia, either in the cytoplasm or the cellular microstructures in comparison with normal women. The intensity of staining of the estrogen receptors on the vaginal neoepithelium of the group of women who underwent the surgical procedure was similar to that of normal women. The estrogen receptors were heterogeneously distributed in the three layers of the tissue in all women. The analysis of the vaginal neoepithelium obtained from amniotic membrane revealed, through the TEM, all the characteristics of a regular vaginal epithelium.

  3. Relação entre a quantidade de AgNORS, atividade proliferativa e o estágio de desenvolvimento placentário em equinos Relationship between the amount of AgNORs, proliferative activity and stage of placental development in horses

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    Ana C.F. Mançanares


    silver nitrate and are related to the activity of rRNA synthesis and to the agility and speed of cell proliferation in the tissues studied. The objective of this study was to relate the amount of AgNORs, proliferative activity and stage of pregnancy in horses, using the coloring of Silver Nitrate. The embryonic attachments were collected, fixed in 10% buffered formaldehyde, embedded in paraplast and stained by silver nitrate. The groups were determined according to the gestational age. The amount of the corium NOR found in early pregnancy indicates the onset of cell activity, and in that the pregnancy progresses, the amount of NOR increases, suggesting higher activity and increased synthesis of their importance in maintaining the fetus. Contrary to what occurs in the corium, the quantification of NORs was higher in late pregnancy than in the beginning, suggesting the stabilization of these membranes in late pregnancy. The chorionic girdle and the yolk sac were found in early pregnancy and had lots of NORs, suggesting synthesis function and proliferation in early pregnancy, since their functions is maintenance of the embryo until the complete formation of the true placenta (chorio-allantoic membranes. We conclude that the membranes that develop in a progressive manner in accordance with the growing embryo/fetal (chorion, amnion and allantoic membranes have an increased number of NORs and the membranes that involute after the formation of the embryo/fetus (yolk sac and chorionic girdle have a decrease in number, suggesting a reduction in proliferative activity in these membranes.

  4. 妊娠合并肺动脉高压23例妊娠结局分析%Pregnancy outcome analysis of 23 pregnant women with pulmonary arterial hypertension

    Institute of Scientific and Technical Information of China (English)



    Objective To investigate the prevention method and mother and fetus outcomes of pregnant women with pulmonary arterial hypertension (PAH).Methods The clinical data of 23 pregnant women with PAH who treated in the first people' s hospital of Shangqiu from June 2003 to May 2013were retrospectively analyzed.System analysis was taken on their PAH,cardiac failure occmrence,pregnancy termination method and mother and fetus outcomes.Results Among the 23 cases of patients,there were 8 cases with mild PAH,9 cases with moderates PAH and 6 cases with serious PAH.Fourteen cases(60.9%) with cardiac failure,the cardiac failure occurrence rate of the patients with moderates and serious PAH was obviously higher than that of the patients with mild PAH (P < 0.05).Two cases of midtrimester pregnancy induced labor with rivanol injection in their amnion cavity,5 cases of midtrimester pregnancy with unmanageable cardiac failure induced labor with cesarean section,3 cases of patients with mild PAH were taken vaginal delivery method under the closely monitoring,13 cases were taken continuous epidural anesthesia cesarean section.Seven perinatal infants were iatrogenic induced labour,12 cases were premature delivery,4 cases were partus matures.Seven newborns occurred neonatal asphyxia and three of them died,in the total 23 cases of the pregnant and lying-in women,4 cases died,19cases left the hospital for their cardiac function improved.Conclusions Pregnant woman with PAH should take early diagnosis and exact risk assessment,improve the patients' cardiac function,strengthen antenatal examination and monitoring measures,terminate pregnancy in time,which can improve the patients' prognosis and get better pregnancy outcome.%目的 探讨妊娠合并肺动脉高压患者的防治方法及母婴结局.方法 回顾性分析2003年6月至2013年5月间收治的23例妊娠合并肺动脉高压患者的临床资料,分析肺动脉高压及心力衰竭发生、妊娠终止方法

  5. 不同方法在唐氏综合征产前筛查中的比较研究%Comparative study of different methods in Down's syndrome prenatal screening

    Institute of Scientific and Technical Information of China (English)

    欧秀月; 黄劲柏


    Objective:To comparison of different period of two CRC screening with the traditional two CRC screening in second trimester prenatal for Down's syndrome in effect. Methods:Using the Roche electrochemical detection technology,1289 cases accepted antenatal screening of pregnant women and5 patients who had been diagnosed with Down syndrome pregnancy of cryopreserved serum specimens in gestational weeks11 to14weeks of pregnancy determination of PAPP-A, Free beta-HCG, and then in 16~20 weeks pregnancy detection of AFP, HCG, concentration, were calculated for two screening and triple screening for risk value, the detection rate and false positive rate.Results:The study of early pregnancy detection of Down syndrome in 21 cases of high risk pregnant women, the second trimester detection of high risk pregnant women in 31 cases, including a period of high risk pregnant women in 20 cases, visible, there were32 cases of high risk pregnant women,31 cases underwent amnion cavity puncture examination, prenatal diagnosis in 1 cases, the positive rate is 3.2%. In 32 cases the screening case,30~40 years old accounted for 16 cases (50%), visible in women of advanced maternal age is down syndrome risk population.When the cut-off value of 1 to 270, with two triple screening for CRC screening for Down syndrome detection rates were65.2%;when the cut-off value of 1:380, different period two CRC screening detection rate was significantly higher than that of the traditional two CRC screening in( P<0.05) .Conclusions:In pregnant women during different periods of the traditional two CRC screening in second trimester prenatal is more effective for Down syndrome screening method, it is necessary to develop the region for the risk of cutting value, in order to improve prenatal screening efficiency.%目的:比较不同时期二联筛查与传统孕中期二联筛查在唐氏综合征产前筛查中的效果。方法:采用罗氏电化学检测技术,对1289例同意

  6. Urocortin在人类妊娠中的定性研究%Characterization of urocortin in human pregnancy

    Institute of Scientific and Technical Information of China (English)


    Objective To examine whether urocortin is produced locally to regulate utero-placental vascular tone during pregnancy. Methods We examined the distribution of urocortin in human placenta, fetal membranes and uterine tissue at term in the presence and absence of labor using a urocortin antibody produced in our laboratory and the immunoperoxidase staining method. Subsequently, we tested urocortin secretion from chorio-decidual cells in vitro using an immunoblot technique. Then, we tested whether urocortin is present in maternal plasma throughout gestation using a radioimmunoassay. A Sephadex G-50 column was used to examine whether immunoreactive urocortin (IR-urocortin) in maternal plasma is the same as synthetic urocortin. Results IR-urocortin was observed in vascular smooth muscle of myometrium decidual stromal cells, syncytiotrophoblast and amnion epithelium. No differences in staining intensity for urocortin were detected between tissues obtained in the absence or presence of labor. Staining intensity for IR-urocortin was greatest in the decidua, suggesting this may be the main site of urocortin production. Positive staining for urocortin was observed in 40% of chorio-decidual cells with 34% of these cells secreting urocortin under basal conditions. Urocortin was detectable in maternal plasma from 16 weeks gestation and concentrations did not change as gestation progressed. IR-urocortin in the maternal plasma eluted from a Sephadex G-50 column at the same site as synthetic urocortin and had a calculated retention co-efficient of 0.44. Conclusion This study indicates that urocortin is produced by the decidua during human pregnancy and is detectable in maternal plasma. These data are consistent with the hypothesis that urocortin is produced locally by the decidua and may act to regulate utero-placental blood flow.%目的 Urocortin是最新发现的促肾上腺皮质激素释放激素(corticotropin releasing hormone,CRH)肽类家族在人类的一

  7. Neural Differentiation of Human Umbilical Cord Mesenchymal Stem Cells by Cerebrospinal Fluid

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    Shirin FARIVAR*


    chick embryos. J Exp Zool A Comp Exp Biol 2004 Apr 1;301(4:280-9.Mitchell KE, Weiss ML. Matrix cells from Wharton’s jelly form neurons and glia. Stem Cells 2003;21(1:50-60.Marcus AJ, Woodbury D. Fetal stem cells from extra-embryonic tissues: do not discard. J Cell Mol Med 2008 Jun;12(3:730-42. doi: 10.1111/j.1582- 4934.2008.00221.x. Epub 2008 Jan 11.Miao Z, Jin J, Chen L, Zhu J, Huang W, Zhao J, Quian H, Zhang X. Isolation of mesenchymal stem cells from human placenta: comparison with human bone marrow mesenchymal stem cells. Cell Biol Int 2006 Sep;30(9:681-7. Epub 2006 Apr 22.In ‘tAnker PS, Scherjon SA, Kleijburg-van der Keur C, Noort WA, Claas FHJ, Willemze R, Fibbe WE, Kanhai HHH. Amniotic fluid as a novel source of mesenchymal stem cells for therapeutic transplantation. Blood 2003;102(4:1548-49.Magatti M, De Munari S, Vertua E, Gibelli L, Wengler GS, Parolini O. Human amnion mesenchyme harbors cells with allogeneic T-cell suppression and stimulation capabilities. Stem Cells 2008 Jan;26(1:182-92. Epub 2007 Sep 27.Kang XQ, Zang WJ, Bao LJ, Li DL, Xu XL, Yu XJ. Differentiating characterization of human umbilical cord blood-derived mesenchymal stem cells in vitro. Cell Biol Int 2006 Jul;30(7:569-75. Epub 2006 Mar 6.Kern S, Eichler H, Stoeve J, Kluter H, Bieback K. Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells 2006 May;24(5:1294-301. Epub 2006 Jan 12.Wagner W, Wein F, Seckinger A, Frankhauser M, Wirkner U, Krause U, et al. Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood. Exp Hematol 2005 Nov;33(11:1402-16.Jackson JS, Golding JP, Chapon C, Jones WA, Bhakoo KK: Homing of stem cells to sites of inflammatory brain injury after intracerebral and intravenous administration: a longitudinal imaging study. Stem Cell Res Ther 2010 Jun 15;1(2:17. doi: 10.1186/scrt17.Romanov YA, Svintsitskaya VA, Smirnov VN. Searching for alternative