ObjectiveTo identify the differential expressed proteins,and to investigate the relationship between altered expression of annexin A4 during window of implantation [ WOI ( at day-6 after ovulatory day )] in infertile patients with endometriosis and endometrial receptivity.MethodsTwo-dimensional fluorescence differential in-gel electrophoresis (2D-DIGE) and matrix-assist laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS) were used to detect protein expression in endometrial WOI in 10 infertile cases with endometriosis as endometriosis group and 10 infertile cases with tubal factors as control group.The semi-quantitative validation of annexin A4 in the eutopic endometrial tissue during WOI was analyzed by western blot.Results By comparing protein profiles,there were 7 meaningful differential proteins during WOI in infertile patients with endometriosis.One protein with an isoelectric point of 5.84 and relative molecular weight of 36 100 were down regulated 348％ in samples of endometriosis group.It was identified as annexin A4 by mass spectrometry.By western blot,relative intensity of annexin A4 in endometriosis group was 7.2 ±0.9,which was lower than 17.8 ± 2.6 in control group significantly (t =7.654,P =0.002 ).ConclusionLower expresssion of annexin A4 during WOI in infertile patients with endometriosis might be associated with the decrease of endometrial receptivity.

Objective To investigate the value of ratio of luteinizing hormone (LH) to folliclestimulating hormone (FSH) in diagnosis of polycystic ovarian syndrome (PCOS) among women with ploycystic ovary (PCO) and to compare the difference of the diagnostic criteria between the Rotterdam Consensus and the Committee for Reproductive and Endocrine in Japan Society of Obstetrics and Gynecology.Methods By means of transvaginal Doppler ultrasound, 195 women with PCO were diagnosed in Nanfang Hospital of Reproductive Medicine Center and compare difference of multiple clinical indexes according to Rotterdam consensus and Japan consensus respectively. In the mean time, the ratio of LH/FSH, the level of LH, testosterone (T) and recevier operating characteristic (ROC) curve were explored to on the value of diagnosis of PCOS. Results By Rotterdam consensus, 144 women were diagnosed with PCOS and 51 women were non-PCOS, while 111 were identified as PCOS and 84 were non-PCOS according to Japan consensus. LH/FSH in PCOS and non-PCOS were 1.59 ±0. 84 and 0. 85 ±0. 47 respectively when based on Rotterdam consensus, and this ratio were 1.87 ± 0. 76 in PCOS and 0. 78 ± 0. 39 in non-PCOS based on Japan consensus. When using LH/FSH to diagnosis PCOS by Rotterdam consensus and Japan consensus,areas under ROC curve are 0. 786 and 0. 942, respectively. Conclusions The ratio of LH/FSH ≥ 1 provide the significant value in the diagnosis of PCOS. The criteria of the Committee for Reproductive and Endocrine in Japan Society of Obstetrics and Gynecology is more suitable for Chinese patients.

OBJECTIVE: To investigate the role of focal adhesion kinase (FAK) in extracellular signal-regulated kinase (ERK) signaling pathway mediated invadsion of trophoblasts. METHODS: We established a human extravillous cytotrophoblasts in vitro invasion model. Different concentrations of herbimycin A(FAK inhibitor)and PD98059 (ERK inhibitor) were given to observe the influence on the growth of trophoblast cells, FAK, ERK phosphorylation, and trophoblast invasion abilities. RESULTS: The expression of phosphorylated FAK in the extravillous cytotrophoblasts (EVCT) was inhibited by herbimycin A in a concentration-dependent manner and expression of phosphorylated ERK1/2 was also partially reduced. PD98059 had no effect on the expression of phosphorylated FAK. Herbimycin A and PD98059 suppressed the in vitro invasion of EVCT to various degrees. CONCLUSION ERK signaling pathway may be the common pathway for many invasive signals,and play a key role in the regulation of trophoblast invasion.
Benzoquinones ; pharmacology ; Cell Division ; physiology ; Cell Movement ; physiology ; Extracellular Signal-Regulated MAP Kinases ; antagonists & inhibitors ; metabolism ; Flavonoids ; pharmacology ; Focal Adhesion Protein-Tyrosine Kinases ; antagonists & inhibitors ; metabolism ; Humans ; Lactams, Macrocyclic ; pharmacology ; Phosphorylation ; Rifabutin ; analogs & derivatives ; Signal Transduction ; physiology ; Trophoblasts ; cytology ; physiology

Objective To establish an animal model of PCOS and to detect the DNA methylation states of LHR, INSR and AR genes. Methods 24-days-old female Sprague-Dawley (SD) rats were randomly divided into two groups. The rats in the experimental group were given subcutaneous implanting of levonorgestrel silica gel staff and injected 1.5 IU hCG twice daily for 9 days from the 4th day. The rats in the control group were injected with normal saline at the same time. Ovarian morphologic changes, sex hormone levels, fasting serum insulin and glucose were detected. The LHR, INSR and AR genes' DNA methylation patterns were checked by methylation specific PCR in modeling group and control group. Results The ovarian weight and volume in modeling group were higher than those in control group. The ovaries in modeling group showed multiple follicular cysts, and the number of theca cells and interstitial cells increased. Less developed follicles and corpus lutea were seem. The serum level of progesterone, testosterone, luteinizing hormone, fasting insulin and glucose were significantly higher in experimental group than those in control group, so as the LH/FSH ratio and HOMA index. No methylation of LHR and AR genes were found in both groups. The methylation frequency of INSR gene (76.7%) was significantly higher in modeling group than that in control group (P<0. 001). Conclusion The depression of INSR gene's transcriptional induced by DNA methylation is important in the development of insulin resistance in PCOS.

Objective To establish an animal model of PCOS and to detect the DNA methylation states of LHR,INSR and AR genes. Methods 24-days-old female Sprague-Dawley (SD) rats were randomly divided into two groups. The rats in the experimental group were given subcutaneous implanting of levonorgestrel silica gel staff and injected 1.5 IU hCG twice daily for 9 days from the 4th day. The rats in the control group were injected with normal saline at the same time. Ovarian morphologic changes,sex hormone levels,fasting serum insulin and glucose were detected. The LHR,INSR and AR genes' DNA methylation patterns were checked by methylation specific PCR in modeling group and control group.Results The ovarian weight and volume in modeling group were higher than those in control group. The ovaries in modeling group showed multiple follicular cysts,and the number of theca cells and interstitial cells increased. Less developed follicles and corpus lutea were seem. The serum level of progesterone,testosterone,luteinizing hormone,fasting insulin and glucose were significantly higher in experimental group than those in control group,so as the LH/FSH ratio and HOMA index. No methylation of LHR and AR genes were found in both groups. The methylation frequency of INSR gene (76.7%) was significantly higher in modelinggroup than that in control group (P

8 oocytes) according to the number of oocytes collected in one ovary. On day of human chorionic gonadotrophin injection, PSV, EDV, PI, RI, S/D of ovarian stromal artery in the ovarian were detected and perifollicular vascularity were graded with color Doppler ultrasonography. Results Not only PSV and EDV of stromal artery but also perifollicular vascularity in low-response group was significantly lower than that of normal-response and high-response groups. PSV and EDV of ovarian stromal artery and perifollicular vascularity were highly interrelated with ovarian response. Conclusions The increase of PSV and EDV of ovarian stromal artery and perifollicular vascilarity indicate the improvement of perfusion in ovary and ovarian response.

Objective　To investigate the effects of leptin on steroidogenesis of human luteinized granulosa cell in vitro. Methods　Human luteinized granulosa cells were isolated from follicular fluid obtained during oocyte retrieval of in vitro fertilization-embryo transfer program and were cultured with M199 medium plus various concentration of leptin (0, 10, 30, 100, 300 ng/ml)，human menopausal gonadotropin (hMG, 0, 0.1, 0.2, 0.5, 1, 2, 5, 10 IU/ml) and testosterone 100 μg/ml. For 2 days the media were collected for estradiol and progesterone measurements. Results　Addition of leptin alone did not alter estradiol and progesterone production (P>0.05) by human luteinized granulosa cells. Leptin of 10～30 ng/ml concentrations caused a dose-dependent inhibition of estradiol production (P<0.05) while greater than 0.5 IU/ml of hMG were added. There was no effect of leptin on hMG -stimulated progesterone production (P>0.05). Conclusion　Leptin can directly inhibit hMG-stimulated estradiol production by human luteinized granulosa cells in vitro, but has no effect on progesterone production. Leptin may play an important role in follicle development and luteinization.

To show the protective effect of macrophage colony-stimulating factor (M-CSF) on macrophages under oxidative stress,we investigated the effect of M-CSF on RAW264.7 cells incubated with tert-butyl hydroperoxide (tbOOH) using L929 cell conditioned medium (L929-CM) as the source of M-CSF.The results showed that M-CSF could alleviate the tbOOH- induced oxidative injury to RAW264.7 cells.We also found that M-CSF could improve superoxide dismutase (SOD) activity in the cells.MnSOD mRNA expression was also shown to be increased by RT-PCR technique,and the induction could be blocked by actinomycin D.So,we concluded that M-CSF could protect RAW264.7 cells from oxidative injury through inducing MnSOD expression.

Objective To study the expression of HOXA10 gene in the endometrium of normal fertile women and patients with unexplained infertility during different phases of menstrual cycle. Methods Endometrium samples were obtained by curettage in 52 normal fertile women and 38 patients with unexplained infertility during different phases of menstrual cycle, HOXA10 mRNA expression were detected by in situ hybridization and reverse polymerase chain reaction (RT-PCR). Results (1) HOXA10 mRNA were detected in the glandular and stromal cells of endometrium of fertile women during the menstrual cycle. By in situ hybridization (positive unite, PU), HOXA10 mRNA levels were significantly higher in the mid-secretory phase [glandular cells (G) 5.69?0.57,stromal cells (S) 7.48?0.67] and late-secretory phase(G 5.99?0.40,S 7.98?1.08) than those in late proliferative phase (G 3.35?0.20,S 3.20?0.37) and early secretory phase (G 3.07?0.26,S 3.18?0.27)(P

Objective To observe the role of calcitonin (CT) during the implantation. Methods Human endometrial epithelial cells were cultured. After stimulated with various concentrations of CT, intracellular calcium(Ca 2+)in the epithelial and stroma cells and pre-embryos were measured by the laser scanning confocal microscope. Resutls When stimulated with different concentrations of CT, mean fluorescence levels in the epithelial cells were similar to that of the control. However CT can improve intracellar Ca 2+ of preimplantation embryos and stroma cells in a dose-dependent manner and significantly higher than those of controls. When 10 nmol/L CT was added to the culture medium, the intracellular Ca 2+ concentration in 8-cell embryos rose immediately. Embryo exposure to CT was followed by a series of Ca 2+ bursts that persisted for at least 2 hours. No change in Ca 2+ was observed when culture medium alone was added to the embryos. Pre-loading embryos and stoma cells with the Ca 2+ chelator,prevented the increased fluorescence after CT addition. Conclusions CT play an important role during the procesess of implantation. It maybe improve intracellar of preembryos and accerate the development of preembryos.