Mesenchymal stem cells-derived exosomes improve pregnancy outcome through inducing maternal tolerance to the allogeneic fetus in abortion-prone mating mouse.

Recurrent pregnancy loss (RPL) is three or more times of consecutive spontaneous loss of pregnancy. The underlying cause is complicated and the etiology of over 50% of RPL patients is unclear. In the present study, bone marrow mesenchymal stem cells were isolated from CBA/J female mice and exosomes were isolated from cell culture medium by ultracentrifugation. CBA/J female mice were paired with male DBA/2 to generate abortion prone mouse model, and CBA/J females paired with male BALB/c mice were used as control. Exosomes were injected through uterine horns into pregnant CBA/J mice on day 4.5 of gestation in abortion-prone matting. On day 13.5 of pregnancy, abortion rates were calculated and the level of transforming growth factor-ß (TGF-ß), interleukin 10 (IL-10), interferon g (IFN-γ), and tumor necrosis factor a (TNF-α) in CD4+ T cells and macrophages in deciduas were evaluated by flow cytometry. Exosomes injection improved the pregnancy outcomes in abortion prone mice. The IL-4 and IL-10 levels on CD4+ T cells were upregulated in the maternal-fetal interface; meanwhile, the TNF-α and IFN-γ levels on CD4+ T cells were reduced. The IL-10 level was increased and IL-12 was reduced on the monocytes that separated from deciduas. miR-101 level was increased in the CD4+ T cells in the deciduas. In conclusion, the treatment of ESCs-derived exosomes modulates T cells' function and macrophages activities in the maternal-fetal interface that resulted in a decreased embryo resorption rate, and provides a therapeutic potential to treat RPL.

Effects of sperm proteins on fertilization in the female reproductive tract.

Mammalian fertilization that culminates by fusion of the male and female gametes is intricately regulated within the female reproductive tract. To become competent to fertilize an egg, the mammalian spermatozoa that enter the female reproductive tract must undergo a series of physiological changes, including hyperactivation, and capacitation. For reaching full competency, the acrosome, a specialized membrane-bound organelle that covers the anterior part of the sperm head, must undergo an acrosome reaction. For becoming competent to bind an ovum, and to penetrate the zona pellucida and cumulus, many sperm proteins are released in the course of the acrosome reaction. Ultimately, the acrosome binds to the oolemma and fusion of sperm and egg occurs. In this review, we outline current understanding of the roles and effects of some essential sperm proteins and their functions during fertilization in the female reproductive tract.

Recent advances in the regulation of testicular germ cell tumors by microRNAs.

Testicular germ cell tumors (TGCTs) are generally rare but represent the most common solid tumors in young men. They are classified broadly into seminoma, which resemble primordial germ cells (PGCs), and non-seminoma, which are either undifferentiated (embryonic carcinoma) or differentiated (teratoma, yolk sac tumor, choriocarcinomas) patterning. A widespread role for microRNAs (miRNAs), in diverse molecular processes driving initiation and progression of various types of TGCTs has been recently studied. We discuss the involvement of different miRNAs in the development and progression of different types of TGCTs. Moreover, we highlight the aberrant expression of miRNAs in TGCTs and several targets, which may define miRNAs as oncomiRs or tumor suppressors. A better understanding of miRNA biology may ultimately yield further insight into the molecular mechanisms of tumorigenesis and new therapeutic strategies against TGCTs.

Regulation of follicular development and differentiation by intra-ovarian factors and endocrine hormones.

Primordial germ cell migration and homing within the gonadal ridge during early embryo development requires oocyte-secreted polypeptide, growth factors, growth and differentiation factors (GDFs), bone morphogenetic proteins, stem cell factor (SCF), and basic fibroblast growth factor (bFGF). During embryogenesis, the germ cells migrate into developing gonads and undergo intra-ovarian development which involves the contact of primordial germ cells with other cells. Further follicular development and differentiation is tightly regulated by hormones and by intraovarian regulators. Maturation of cumulus-oocyte complexes and ovulation are directly controlled by FSH and LH and requires activation of mitogen-activated protein kinase in granulosa cells. The selection of dominant follicles is driven by a series of proliferation and apoptotic events. Together, the available data suggests that follicular development is regulated both by systemic and local factors.

Effect of Acupuncture and Clomiphene in Chinese Women With Polycystic Ovary Syndrome: A Randomized Clinical Trial.

Importance: Acupuncture is used to induce ovulation in some women with polycystic ovary syndrome, without supporting clinical evidence. Objective: To assess whether active acupuncture, either alone or combined with clomiphene, increases the likelihood of live births among women with polycystic ovary syndrome. Design, Setting, and Participants: A double-blind (clomiphene vs placebo), single-blind (active vs control acupuncture) factorial trial was conducted at 21 sites (27 hospitals) in mainland China between July 6, 2012, and November 18, 2014, with 10 months of pregnancy follow-up until October 7, 2015. Chinese women with polycystic ovary syndrome were randomized in a 1:1:1:1 ratio to 4 groups. Interventions: Active or control acupuncture administered twice a week for 30 minutes per treatment and clomiphene or placebo administered for 5 days per cycle, for up to 4 cycles. The active acupuncture group received deep needle insertion with combined manual and low-frequency electrical stimulation; the control acupuncture group received superficial needle insertion, no manual stimulation, and mock electricity. Main Outcomes and Measures: The primary outcome was live birth. Secondary outcomes included adverse events. Results: Among the 1000 randomized women (mean [SD] age, 27.9 [3.3] years; mean [SD] body mass index, 24.2 [4.3]), 250 were randomized to each group; a total of 926 women (92.6%) completed the trial. Live births occurred in 69 of 235 women (29.4%) in the active acupuncture plus clomiphene group, 66 of 236 (28.0%) in the control acupuncture plus clomiphene group, 31 of 223 (13.9%) in the active acupuncture plus placebo group, and 39 of 232 (16.8%) in the control acupuncture plus placebo group. There was no significant interaction between active acupuncture and clomiphene (P = .39), so main effects were evaluated. The live birth rate was significantly higher in the women treated with clomiphene than with placebo (135 of 471 [28.7%] vs 70 of 455 [15.4%], respectively; difference, 13.3%; 95% CI, 8.0% to 18.5%) and not significantly different between women treated with active vs control acupuncture (100 of 458 [21.8%] vs 105 of 468 [22.4%], respectively; difference, -0.6%; 95% CI, -5.9% to 4.7%). Diarrhea and bruising were more common in patients receiving active acupuncture than control acupuncture (diarrhea: 25 of 500 [5.0%] vs 8 of 500 [1.6%], respectively; difference, 3.4%; 95% CI, 1.2% to 5.6%; bruising: 37 of 500 [7.4%] vs 9 of 500 [1.8%], respectively; difference, 5.6%; 95% CI, 3.0% to 8.2%). Conclusions and Relevance: Among Chinese women with polycystic ovary syndrome, the use of acupuncture with or without clomiphene, compared with control acupuncture and placebo, did not increase live births. This finding does not support acupuncture as an infertility treatment in such women. Trial Registration: clinicaltrials.gov Identifier: NCT01573858.

Periodic elevation of regulatory T cells on the day of embryo transfer is associated with better in vitro fertilization outcome.

Treg cells have been shown to be important in maintaining maternofetal tolerance, but the expression of Tregs in assisted reproductive technology (ART) in women on the day of embryo transfer (D0), 5days (D5) and 14days after ET (D14); the related factors influencing the expression levels of Tregs; the proliferation ability and the relevant cytokine epression by Tregs on D14 have not been investigated. In this study, 124 women undergoing in vitro fertilization-intracytoplasmic sperm injection (IVF/ICSI) were enrolled. Early morning fasting blood samples were obtained for the measurement of Tregs and other relevant indicators on the D0, D5and D14days after ET. we showed that the Tregs were increased on D0 and D14 in pregnant women, while there was no obvious fluctuation in non-pregnant women. IL-10 and TGF-ß levels and the expansion of Tregs were significantly higher in successfully pregnant women than in non-pregnant women on D14. The levels of E2, P did not significantly differ between the groups. We suggest that periodic elevation of Tregs on the day of ET was associated with higher embryo implantation rate after ART.

Effect of PDGF-Rb antagonist imatinib on endometrial injury repairing in mouse model.

OBJECTIVE: To study the effects of PDGF-Rb antagonists imatinib on endometrial injury repairing in the mouse model. METHODS: The cultured MSCs cells from male mice were marked with BrdU in vitro, and then transplanted to the female mice which suffered from radiation injury through tail vein, PDGF-Rb antagonists imatinib was injected through abdominal cavity. Four groups were arranged, which were radiation transplantation group, normal control group, imatinib intervention group and radiation control group. BrdU incorporation, SRY expression and MVD status were detected in uterus of mice. RESULTS: SRY gene was negative expressed in normal control group and radiation control group. SRY gene presented positive in radiation transplantation group and imatinib intervention group; BrdU incorporation showed negative in radiation control group and normal control group which died in the early stage in mice; the incorporation of BrdU was higher in radiation transplantation group compared with imatinib intervention group; CD34 was positive on the uterus of all the four groups, which showed highest in radiation control group and lowest in radiation control group; The MVD in imatinib intervention group was lower than radiation control group; the difference of MVD was significantly compared with normal control group (P < 0.05). CONCLUSIONS: PDGF-Rb antagonists imatinib could inhibit the repairing function of MSCs in the endometrial lesions in mice.

Clinical application of operative hysteroscopy in treatment of complex hydrosalpinx prior to IVF.

BACKGROUND: In vitro fertilization and embryo transfer (IVF-ET) is the best option for patients with hydrosalpinx. However, if hydrosalpinges is not pre-treated, the therapeutic outcomes of IVF-ET would be compromised. OBJECTIVE: This study aims to investigate the safety and effects of operative hysteroscopy in the treatment of patients with hydrosalpinx prior to IVF-ET, who were not indicated for laparotomy due to extensive pelvic adhesion. MATERIALS AND METHODS: The study analyses retrospectively data from 10 women with hydrosalpinx, who were unable to undergo laparotomy due to extensive pelvic adhesion and treated by operative hysteroscopy prior to IVF-ET, and was assessed the effects and safety of the procedure. RESULTS: Postoperative Hystero-salpingography demonstrated complete tubal occlusion of the diseased side in all cases. Being applied with IVF-ET for fertility after their hysteroscopy operation, 5 out of 10 patients acquired clinical pregnancy. CONCLUSION: Hysteroscopic tubal occlusion of the proximal part of the hydrosalpinx can effectively prevent the hydrops backflow to endometrial cavity and benefit subsequent implantation in the course of assisted reproduction without significant complications.

Adoptive transfer of pregnancy-induced CD4+CD25+ regulatory T cells reverses the increase in abortion rate caused by interleukin 17 in the CBA/JxBALB/c mouse model.

STUDY QUESTION: Could adoptive transfer of pregnancy-induced CD4+CD25+ regulatory T cells (Tregs) reverse the increase in abortion rate caused by interleukin 17 (IL-17) in the CBA/J × BALB/c mouse model? SUMMARY ANSWER: The effects of exogenous IL-17 on increased abortion rate, as well as decreased transforming growth factor (TGF)-ß and IL-10 expression, are reversed by a pre-mating transfusion of Tregs in a mouse model of pregnancy. WHAT IS KNOWN ALREADY: IL-17 is a pro-inflammatory cytokine mainly expressed by T helper 17 cells, and plays a pivotal role in the pathogenesis of endometriosis, miscarriage, preterm labor and pre-eclampsia. The activity of Th17 cells is attenuated by the anti-inflammatory action of Tregs. STUDY DESIGN, SIZE, DURATION: Fifty microliters of phosphate-buffered saline (PBS) (Group 1,) or recombinant IL-17 (rIL) (10 µg/mouse) supernatant (Group 2) was administered in the vaginal vaults of anesthetized pregnant CBA/J mice on Day 1 of pregnancy. Tregs (2 × 10(5) cells) purified from pregnant CBA/J × BALB/c mice were given i.v. via the tail vein 2 days before mating (Group 3) or on Day 7 of pregnancy (Group 4). PARTICIPANTS/MATERIALS, SETTING, METHODS: Mice (n = 40) were randomly assigned to one of four experimental groups. The numbers of surviving and reabsorbed fetuses in each group were counted on Day 14 of pregnancy, and the expression of interferon (IFN)-γ, IL-4, TGF-ß and IL-10 in the decidual tissue was assessed by real-time RT-PCR and western blotting. MAIN RESULTS AND THE ROLE OF CHANCE: Normal pregnant CBA/J mice mated with BALB/c males which received transvaginal rIL-17 presented with a significantly increased abortion rate compared with the group which received PBS (27.7 versus 9.9%, respectively; P < 0.05). The transfusion of pregnancy-induced Tregs from 14-day normal pregnant mice 2 days before mating reduced the abortion rate caused by IL-17 (12.5 versus 27.7%, respectively; P < 0.05), while transfusion of Tregs on Day 7 of pregnancy had no effect. Transfusion of Tregs did not affect IFN-γ or IL-4 expression in the decidual tissue at either the mRNA or protein level. Administration of rIL-17 resulted in a decrease in production of TGF-ß and IL-10 at both mRNA and protein levels (P < 0.05). Transfusion of Tregs before mating increased TGF-ß and IL-10 mRNA and protein levels (P < 0.05), while Tregs transfusion at Day 7 of pregnancy had no effect on TGF-ß or IL-10 expression. LIMITATIONS, REASONS FOR CAUTION: These data derive from only a small number of mice. It is unclear whether the same effects would be seen in humans. WIDER IMPLICATIONS OF THE FINDINGS: Abnormally elevated expression of IL-17 in the feto-maternal interface may result in miscarriage. Transfer of antigen-specific Tregs before mating takes place may have potential applications in the prevention of recurrent spontaneous abortion. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by a grant from the National Natural Science Foundation of China (81370013, 81000277 and 81300533) and Shandong Provincial Natural Science Foundation, China (ZR2013HQ002). There were no conflicts of interest.

Regulation of the expression of Th17 cells and regulatory T cells by IL-27 in patients with unexplained early recurrent miscarriage.

In normal pregnancy, tolerance of the maternal immune system with regard to the genetically incompatible fetus depends on the interactions of an array of cytokines secreted by maternal and fetal cells at the site of implantation. Earlier research indicating that altered immunity exists in unexplained recurrent miscarriage (RM) has been dominated by the Th1/Th2 hypothesis. Recently, the Th1/Th2 paradigm has been expanded into the Th1/Th2/Th17 and regulatory T cells paradigm. We recently demonstrated a prevalence of Th17 cells, an inverse relationship between Th17 cells and regulatory T cells and deregulation of Th17 cells by regulatory T cells in early pregnancy in unexplained RM patients. In this study, we investigated the expression of IL-27 and the role of the cytokine IL-27 in the regulation of Th17/Treg expression. Quantitative real-time RT-PCR and Western blot analyses were performed to evaluate IL-27 expression in deciduas from unexplained RM patients, spontaneous miscarriage (SM) patients and healthy women following elective abortion in the early stages of normal pregnancy (control). Regulation of IL-17, TGF-ß and IL-10 expression in CD4(+) T cells in unexplained RM patients by IL-27 was assessed using enzyme-linked immunosorbent assay (ELISA). Expression of IL-27 was lower in deciduas of patients with unexplained RM compared with SM and control subjects. IL-27 inhibited IL-17 expression and enhanced IL-10 expression in a dose-dependent manner. IL-27 had no effect on TGF-ß expression. IL-27 regulates the expression of IL-17 and IL-10, which are predominantly secreted by Th17 cells and regulatory T cells in unexplained RM patients.

Prediction of IVF/ICSI outcome based on the follicular output rate.

This study assessed the true accuracy of follicular output rate (FORT) as a prognostic indicator of response to FSH and reproductive competence after IVF/intracytoplasmic sperm injection. A total of 1643 cycles, including 140 polycystic ovary syndrome (PCOS) patients who underwent ovarian stimulation, were studied. FORT was calculated as the ratio of preovulatory follicle count on the day of stimulation×100/small antral follicle count (3-10mm in diameter) at baseline. Low, medium and high FORT groups were defined according to tertile values. Among 1503 non-PCOS cycles, numbers of retrieved oocytes and of all embryos that could be transferred, as well as rates of good-quality embryos, embryo implantations and clinical pregnancies, progressively increased with FORT. In PCOS patients, FORT were significantly lower in patients who achieved clinical pregnancy compared with those who did not (0.56±0.21 versus 0.66±0.29, P=0.031). Fertilization and good-quality embryo rates were significantly higher with medium FORT than low and high FORT (P=0.001 and P=0.047, respectively). Medium FORT in PCOS patients and high FORT in non-PCOS patients may predict better outcomes for IVF/ICSI.

Dysregulation of macrophage activation by decidual regulatory T cells in unexplained recurrent miscarriage patients.

CD4(+)CD25(+) T cells (Treg cells) and macrophages play roles in the maintenance of maternal-fetal immunological tolerance. Treg cells suppress the function of macrophages via mechanisms mediated by cell-cell contact and production of soluble factors. The purpose of this study was to investigate regulation of macrophages by Treg cells within decidua from patients with unexplained recurrent miscarriage (RM) and normal control women during early pregnancy. Treg cells and macrophages were isolated from deciduas of unexplained RM (n=15) and control women (n=15) by magnetic cell separation and co-cultured for six days. Regulation of macrophages by Treg cells was assessed in the presence and absence of neutralizing anti-TGFß antibodies and in transwell experiments. Expression of CD80, CD86, IL10, and IFNγ by macrophages was measured by flow cytometry or ELISA. Macrophage expression of CD80 and CD86 was higher in deciduas of unexplained RM patients compared with controls whereas the expression of IL10 was lower. There was no difference in the expression of IFNγ by macrophages between the two groups. Treg cells inhibited macrophage expression of CD80, CD86 and IFNγ and increased the expression of IL10. The regulatory effects of Treg cells were abrogated in the presence of neutralizing anti-TGFß antibodies or by transwell culture. The phenotype of macrophages therefore differed in unexplained RM patients compared with normal early pregnant subjects. Macrophage regulation by Treg cells was shown to be mediated by cell-cell contact and TGFß and this capacity was decreased in unexplained RM patients.

[Clinical efficacy of Chinese herbal retention enema combined with intrauterine douching for patients with endometritis].

OBJECTIVE: To study the clinical efficacy of Chinese herbal retention enema combined with intrauterine douching for patients with endometritis diagnosed by hysteroscopy. They failed in in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI)-embryo transfer (ET) treatment. METHODS: 131 patients received hysteroscopy after they failed in routine IVF/ICSI-ET treatment. Of them, 66 patients diagnosed as endometritis were enrolled as the test group and 65 patients without endometritis were enrolled as the control group. Chinese herbal retention enema combined with intrauterine douching was performed on patients in the test group before the next IVF/ICSI, while direct IVF/ICSI was performed on those in the control group. The embryo implantation rate and the clinical pregnancy rate were compared between the two groups. RESULTS: The clinical pregnancy rate and the embryo implantation rate were 48.5% and 24.2% respectively, while they were 29.2% and 14.9% respectively in the control group, showing significant difference (P < 0.05). CONCLUSION: Chinese herbal retention enema combined with intrauterine douching could improve the embryo implantation rate and the clinical pregnancy in patients with endometritis.

The deregulation of regulatory T cells on interleukin-17-producing T helper cells in patients with unexplained early recurrent miscarriage.

BACKGROUND: CD4(+)CD25(+) regulatory T cells (Tregs) are important for the maintenance of immune homeostasis by virtue of their ability to control T-cell proliferation in the peripheral blood (PB). We recently demonstrated that the prevalence of Tregs is decreased, whereas that of Th17 cells is increased, in the PB and decidua samples of patients with unexplained recurrent miscarriage (RM). In this study, we investigated whether the cytokine production of Th17 cells can be suppressed by the Tregs and elucidated the mechanism by which Tregs exert this suppressive effect. METHODS: Flow cytometry was used to analyze the surface phenotype and cytokine production of Th17 cells in the PB of women with unexplained RM (n = 17) and healthy women in early stages of pregnancy who underwent elective abortion (n = 20). The suppressive ability of Tregs on Th17 cells was assessed in in vitro co-cultures and transwell experiments. The amount of secreted interleukin-17 (IL-17) in the supernatants was measured by enzyme-linked immunosorbent assay (ELISA). The inhibitory activity of transforming growth factor-ß (TGF-ß) and IL-10 on IL-17 expression in CD4(+) T cells was assessed using ELISA. RESULTS: The proportions of IL-17-positive CD4(+) T cells, CC chemokine receptor type 6 (CCR6)-positive CD4(+) T cells and CCR6 expression of IL-17-positive CD4(+) T cells were higher in the PB samples of patients with unexplained RM than in PB of healthy control subjects. In vitro, Tregs could inhibit the expression of IL-17; more Th17 cells were inhibited in the control group than in the unexplained RM group. High-dose TGF-ß inhibited the expression of IL-17, whereas IL-10 inhibited IL-17 expression in a dose-dependent manner. CONCLUSIONS: IL-17 expression can be inhibited by Tregs. The suppressive activity of Tregs on Th17 cells was decreased in patients with unexplained RM. The ability of Tregs to suppress cytokine secretion might be effected by a cell-cell contact. TGF-ß and IL-10 could inhibit the expression of IL-17.

Evaluation of the association between the CYP19 Tetranucleotide (TTTA)n polymorphism and polycystic ovarian syndrome(PCOS) in Han Chinese women.

OBJECTIVE: Evidence indicates that the CYP19 gene is a positional and functional candidate for genetic study in polycystic ovarian syndrome (PCOS). The present study aims to evaluate the association between tetranucleotide TTTA repeat polymorphism in the CYP19 gene and PCOS among Han Chinese women. METHODS: Clinical materials employed in this study consist of 123 patients with PCOS and 113 healthy controls. The CYP19 tetranucleotide TTTA repeat polymorphism was genotyped with a protocol of PCR and fluorescent capillary electrophoresis. RESULTS: Common allele of the CYP19 tetranucleotide TTTA repeat polymorphism in this population of Han Chinese women was 11R. The frequency of 11R in PCOS was lower than in the control subjects (34.55% vs 42.92%, p=0.046). The carriers with allele 11R in PCOS had decreased CHO (5.00+/-0.63 vs 6.14+/-0.85 mmol/L, p=0.012). The carriers with allele 7R-TCT in PCOS had increased CHO (5.96+/-0.83 vs 5.08+/-0.65 mmol/L, p=0.027) and LDL (5.11+/-0.77 vs 4.31+/-0.66 mmol/L, p=0.014) compared to the patients carrying other alleles. CONCLUSIONS: The most common allele of the tetranucleotide TTTA repeat polymorphism in the forth intron of CYP19 gene in Han Chinese women is 11R, which was different with the previous study in European Caucasians. Allele 11R may be associated with PCOS in the population of Han Chinese women, and it may refrain from the hypercholesteremia of PCOS. Allele 7R-TCT may be related to the lipid metabolism of PCOS. This CYP19 tetranucleotide TTTA repeat polymorphism is an ethnic and racial variant and moderately contributes to the pathogenesis of PCOS in the population of Han Chinese women.

Increased prevalence of T helper 17 (Th17) cells in peripheral blood and decidua in unexplained recurrent spontaneous abortion patients.

T helper 17 (Th17) cells and regulatory T (Treg) cells are two distinct subsets of CD4(+) T cells which have opposite effects on inflammation, autoimmunity and immunological rejection of foreign tissue. Treg cells have been shown to be important in maintaining materno-fetal tolerance, but the role of Th17 cells in human pregnancy and pathological pregnancy, especially in relation to unexplained recurrent spontaneous abortion (RSA), has not been investigated. In this study, we showed that the proportion of Th17 cells in the peripheral blood and decidua was significantly higher in unexplained RSA patients compared to normal, early pregnant women. Meanwhile, there was an inverse relationship between Th17 cells and Treg cells in the peripheral blood lymphocytes (PBL) and decidua in unexplained RSA. The expression of Th17 related factors, IL-17, IL-23 and retinoid orphan nuclear receptor (RORC), in PBL and decidua in unexplained RSA patients was significantly higher than normal, early pregnant women. This study is the first to define the occurrence of Th17 cells in unexplained RSA patients and in normal, early pregnant women. We suggest that these highly pro-inflammatory cells contribute to unexplained RSA, and the balance between Th17 cells and Treg cells may be critical to pregnancy outcomes.

Evaluation of association between the CYP11alpha promoter pentannucleotide (TTTTA)n polymorphism and polycystic ovarian syndrome among Han Chinese women.

Evidence indicates that CYP11alpha is a positional and functional candidate for genetic study in polycystic ovarian syndrome (PCOS). In the present study, we have evaluated the association between the CYP11alpha promoter pentannucleotide (tttta)n polymorphism and PCOS among Han Chinese women. Subjects include 125 patients with PCOS and 121 healthy controls, and all were Han Chinese women. Clinical characteristics of patients with PCOS and control subjects were examined according to the Rotterdam consensus criteria. The CYP11alpha promoter pentannucleotide (tttta)n polymorphism was genotyped with PCR and fluorescent capillary electrophoresis protocol. Results indicate that common alleles of the CYP11alpha promoter pentannucleotide (tttta)n polymorphism in this population of Han Chinese women were P4, P6 and P8, and allele P6 was the most common one. Frequencies of those three common alleles between PCOS cases and controls (24.8, 65.6, 6.8% and 23.6, 65.7, 9.9%, respectively) were similar. No significant allelic association of this polymorphism with PCOS was found. However, the carriers with allele P6 among patients with PCOS had increased WHR (0.85+/-0.05 vs 0.82+/-0.08, P=0.039) and decreased AUCG (9.5+/-2.1 vs 11.4+/-2.8, P=0.021) compared to the patients carrying other alleles. Therefore, the most common allele of the CYP11alpha promoter pentannucleotide (tttta)n polymorphism in the population of Han Chinese women is P6, while the most common allele in European Caucasians, as previously reported, is P4. This polymorphism is an ethnic and racial variant, and may have the risk susceptibility in abnormal metabolism of patients with PCOS in Han Chinese women.

[Different types of menstrual cycle and their significance in Chinese women diagnosed with polycystic ovary syndrome according to the Rotterdam consensus criteria].

OBJECTIVE: To analyze the clinical and metabolic characteristics of Chinese women with different menstrual types. All the women were diagnosed with polycystic ovary syndrome according to the Rotterdam consensus criteria. METHODS: A total of 2100 patients were divided into three groups: amenorrhea, oligomenorrhea and regular menstruation. The metabolism and endocrine indices were determined and compared among three groups. RESULTS: (1) The incidences of hirsutism and PCO were obviously higher in the amenorrhea group than in the oligomenorrhea group (P < 0.01). (2) The amenorrhea group had the highest level of serum testosterone while the oligomenorrhea group had the lowest. Moreover, the level of luteinizing hormone (LH) was higher in the amenorrhea group than in the other two groups (P < 0.01). However the ratio of LH/FSH increased significantly in the regular menstruation group (P < 0.01). (3) The value of 2 h blood glucose was the highest in the amenorrhea group (P < 0.01). CONCLUSION: PCOS patients with diverse types of menstrual cycle show different clinical manifestations and metabolic and endocrine characteristics so that the choice of treatment should be individualized.

Association of +45G15G(T/G) and +276(G/T) polymorphisms in the ADIPOQ gene with polycystic ovary syndrome among Han Chinese women.

OBJECTIVE: Polycystic ovary syndrome (PCOS) is frequently associated with insulin resistance (IR) and consequently with increased risk of metabolic disorders. Adiponectin is the most abundant adipocytokine and may play a role in the regulation of insulin sensitivity and IR in PCOS. The aim of the present study was to evaluate the genetic influence of the adiponectin (ADIPOQ) gene polymorphisms in the development of PCOS among Han Chinese women. METHODS: Two single nucleotide polymorphisms (SNPs),+45G15G(T/G) and +276(G/T), in the ADIPOQ gene were genotyped in 120 patients with PCOS and 120 healthy control subjects. All of them were Han Chinese women. RESULTS: Both SNPs were found to be significantly associated with PCOS (P=0.021, odds ratios=1.629, 95% confidence intervals: 1.074-2.469 and P=0.015, 1.576, 1.091-2.279 respectively). In SNP +276(G/T), the allele G was found to be significantly associated with increased fasting insulin levels, homeostasis model assessment to assess IR index, and area under the curve glucose levels, but decreased glucose and insulin ratio in the PCOS patients. Furthermore, the patients carrying genotypes G/G and G/T had significantly decreased levels of serum adiponectin (6.16+/-3.18 plus 5.93+/-3.23 vs 8.96+/-3.21 microg/ml, P=0.030) compared with the patients with genotype T/T. CONCLUSIONS: The present study provides evidence that SNPs +45G15G(T/G) and +276(G/T) in the ADIPOQ gene are associated with PCOS in Han Chinese women. SNP +276(G/T) may contribute to an impact of insulin levels and IR, which are implicated in the susceptibility for PCOS.