Objective To explore the relationship between the changes of serum malondialdehyde(MDA),superoxidation dismutase (SOD) and the level of serum-estriol (E 3), ?-hCG, and expression of estrogen receptors(ER), HCG on placenta of intrahepatic cholestasis of pregnancy(ICP). Methods Thirty-three women with ICP and thirty normal pregnant women were enrolled in study. All elbow vein blood were drawn within half an hour before delivery. The level of SOD, MDA, E 3 and?-hCG were detected. Twenty cases of ICP group and control group were selected randomly and placental tissues were measured by using immunohistochemical method to assess the level of ER and hCG. Results The level of MDA and free E 3 in ICP group were [(6.4?2.1) mol/L],and were [(19?9 ng/ml)] higher than that in control group [(5.2?1.4 mol/L)], and [(14?6 ng/ml)] (P

Objective To study the role of gene Fas and FasL expression in apoptosis of plcenta with ICP. Methods TUNEL (TdT-mediated dUTP nick end labeling) and immuno-hisochemistry methods were used for 31 placental samples with ICP and 31 normal placental samples to detect apoptosis index (AI) and the expression of Fas and FasL in placental tissues. Results AI of cytotrophoblast, syncytotrophoblast, decidual cells and mediate cell(49.09?9.13,t=13.41;46.64?9.77,t=12.16;35.09?9.49,t=8.43;38.74?9.70,t=11.28) in ICP group was significantly higher than that of control group (P

BACKGROUND:In vitro isolation and purity technique of stem cells mostly depends on the identification of cell surface marker,such as monoclonal antibody adherent spreading method,flow cell sorting method and immunomagnetic beads sorting method,but the operation was complicated and the price was high.OBJECTIVE:To observe the biological characteristics of human amniotic fluid-derived embryonic mesenchymal stem cells,which were isolated and cultured using the two-step method.DESIGN,TIME AND SETTING:The opening study was conducted at the Stem Cell Research Room of Xinjiang Medical University from March 2008 to March 2009.MATERIALS:Totally 10 amniotic fluid specimens were obtained from pregnant women who underwent prenatal diagnosis following 16-22 weeks of gestation or voluntarily induced abortion.With ultrasonic guidance,amniocentesis was performed to collect 20-40 mL amniotic fluid.METHODS:Human amniotic fluid-derived embryonic mesenchymal stem cells were isolated and cultured using the two-step method.Amniotic fluid was first centrifuged and incubated till spindle-shape cells were seen,with the presence of flbroblast-tike cell colonies.Supematant was moved to a new 25 cm~2 culture flask for further culture till spindle-shape fibroblast-like mesenchymal stem cell colonies.When 70% confluence,cells were digested,and incubated in α-MEM,supplemented with basic fibroblast growth factor,served as the first passage.MAIN OUTCOME MEASURES:Morphological changes in human amniotic fluid-derived embryonic mesenchymal stem cells of primary culture and subculture were measured.Karyotype,cycle,growth curve and colony formation ability of human amniotic fluid-derived embryonic mesenchymal stem cells were measured.Surface antigen and cytokine were examined using flow cytometry,immunofluorescence and RT-PCR.RESULTS:Human amniotic fluid-derived embryonic mesenchymal stem cells were successfully isolated and subcultured.During metaphase,primarily cultured amniotic fluid cells presented scattered spindle cells and flbroblast-like mesenchymal stem cell colonies every 7 days.Passaged cells completely adhered in 12 hours.Following 1 or 2 days of latent period,cells proliferated rapidly.About 90% confluence was observed following 6 or 7 days of culture.Cell arranged regularly,showing whirlpool-shape,radiated shape.Cells were spindle-shape,with unclear boundary.Chromosome karyotype of human amniotic fluid-derived embryonic mesenchymal stem cells was normal diploid.Growth curve showed "S" shape,but the two-step method reached a peak at (6.1±0.5) days,which was significantly rapid compared with the one-step method (7.2±0.6) days (P=0.035).Flow cytometry analyses showed that P3 cells at S phase took up (14±2.3)% using the two-step method,which was more than the one-step method (9.0±1.4)% (P=0.031).Low-density human amniotic fluid-derived embryonic mesenchymal stem cells were incubated for 7 days prior to cells formed scattered cell colonies.However,colony forming efficiency using the two-step method (15.0±2.3)% were significantly more than the one-step method (10.0±1.8)% (P=0.021).Flow cytometry results showed that human amniotic fluid-derived embryonic mesenchymal stem cells expressed CD44,CD29 and CD105,but were negatively for CD45,CD34,HLA-DR.Immunofluorescence suggested that Oct-4-positive cells were observed in amniotic fluid.However,the proportion of Oct-4-positive cells using two-step method (1.2±0.3)% was significantly greater than the one-step method (0.9±0.2)% (P=0.041).RT-PCR suggested that human amniotic fluid-derived embryonic mesenchymal stem cells obtained using the two methods expressed Oct-4.CONCLUSION:Human multipotent mesenchymal stem cells are present in human amniotic fluid.The two-step culture protocol could be a kind of high performance and simple protocol which may not interfere with the normal prenatal diagnosis procedure.

Objective To investigate the regulatory effects of Zishen Yutai Pills on the expression levels of homeboxA10 (HOXA10) and its downstream target gene empty spiracles homebox 2 (EMX2) in the endometria of ovulation-inducing mice at different implantation stages. Methods Seventy-five estrous female Kunming mice were randomly divided into 5 groups, namely normal group, model group 1, model group 2, treatment group 1, treatment group 2, 15 mice in each group. The model group 1 was given short-term protocol for ovulation induction; the model group 2 was given long-term protocol for ovulation induction; the treatment group 1 was given Zishen Yutai pills (at the dose of 0.4 g/mL) on the basis of the protocol for the model group 1; the treatment group 2 was given Zishen Yutai Pills (at the dose of 0.4 g/mL) on the basis of the protocol for the model group 2; the normal group was given intragastric administration or intraperitoneal injection of the same volume of normal saline. The mRNA and protein expression levels of HOXA10 and EMX2 in mouse uterus were detected by real-time fluorescent quantitative polymerase chain reaction (qPCR) and Western blot method, respectively. Results Compared with the normal group, the mRNA and protein expression levels of HOXA10 were decreased, and the mRNA and protein expression levels of EMX2 were increased in model group 1 and model group 2(P< 0.01). Compared with the corresponding model group 1 and 2, the mRNA and protein expression levels of HOXA10 were significantly up-regulated (P < 0.01) , and the mRNA and protein expression levels of EMX2 were decreased in the treatment group 1 and 2 (P < 0.01), respectively. Conclusion Zishen Yutai Pills may improve mouse endometrial receptivity by up-regulating HOXA10 expression and inhibiting EMX2 expression.

Through summarizing and analyzing the characteristics of elderly infertile people,the problems emer-ging in the assisted reproduction process and the possible ethical problems emerging in the assisted reproduction treatment of elderly patients,this paper explored how to build the ethical path which aimed at elderly pregnancy -assisted people and suitable for Reproductive Center in the First Affiliated Hospital of Xinjiang Medical University. And aiming at the possible ethical problems emerging in the process of assisted reproduction treatment of elderly pa-tients,this paper put forward that it should establish normative ethical working path,to be more convenient to fully conduct ethical supervision and examination in the process of assisted reproduction treatment of elderly patients.

Objective : To investigate the expression of macrophage migration inhibitory factor (MIF) and its effect on autophagy and insulin resistance of ovarian granulosa cells in polycystic ovary syndrome ( PCOS) . Methods: Follicular fluid and ovarian granulosa cells were collected from 40 PCOS patients ( n = 40) and non PCOS patients (control group, n = 20) . PCOS included patients with IR (PCOS with IR, n = 20) and patients without IR (PCOS without IR, n = 20) . The expression of MIF in follicular fluid was detected by ELISA . The ratio of autophagy vacuoles to microtubule⁃associated protein light chain Ⅱ( LC3 Ⅱ) /microtubule⁃associated protein light chain Ⅰ( LC3 Ⅰ)in granulosa cells was observed by transmission electron microscopy and Western blot . Human ovarian granule cell line KNG was cultured in vitro and CCK⁃8 was used to detect the effects of different concentrations of MIF on granule cell activity . KNG cells were divided into four groups: normal culture group ( NC group), MIF group, chloroquine group (CQ group) and MIF + CQ group . The effects of MIF on the expression of autophagy related proteins LC3, autophagy related genes 7(Atg7), ubiquitin binding protein(p62), and insulin signaling pathway related proteins insulin receptor substrate⁃1(ISR⁃1), glucose transporter 4(GLUT4), protein kinase B(Akt) phosphorylation were observed by Western blot, and the effect of MIF on glucose uptake ability of granulosa cells was detected by glucose uptake test . Results : Compared with the control group or PCOS without IR group, MIF expression in follicular fluid and autophagy level of granulosa cells in PCOS with IR group increased (P < 0. 05 or P < 0. 01); In vitro experiments showed that MIF could significantly inhibit the cellular activity of KNG in granulocytes in a concentration⁃dependent manner, in which 100 ng/ml MIF was selected for subsequent relevant experiments; Compared with NC group, LC3Ⅱ/LC3I ratio and Atg7 protein in MIF group and MIF + CQ group increased (P < 0. 05), while p62 protein, IRS⁃1 protein, Akt phosphorylation level, GLUT4 protein expression level and glucose uptake ability decreased ( P < 0. 05 ), while the above autophagy markers in MIF + CQ group were significantly higher than those in MIF group (P < 0. 05), and the protein related to insulin signal transduction and glucose uptake increased (P < 0. 05) . Conclusion MIF may promote IR development in PCOS patients by up⁃regulating the autophagy level of granulosa cells, while inhibiting the autophagy of granulosa cells can improve MIF⁃induced IR.