G protein­coupled estrogen receptor/miR­148a/human leukocyte antigen­G signaling pathway mediates cell apoptosis of ovarian endometriosis.

The focus of the current study was a G protein­coupled estrogen receptor (GPER)/microRNA (miR)­148a/human leukocyte antigen­G (HLA­G) signaling pathway in ovarian endometriosis. Reverse transcription­quantitative polymerase chain reaction was performed to analyze the changes in miR­148a expression. A MTT assay, flow cytometry and caspase­3/9 activity assays were performed to analyze cell proliferation, apoptosis and caspase­3/9 activity levels, respectively. Protein expression was measured using western blot analysis. In tissue samples from healthy controls, and patients with endometriosis and endometriosis­associated ovarian cancer, the expression of miR­148a was lower in in endometriosis and EAOC samples compared with healthy controls. Overexpression of miR­148a using miR mimics significantly decreased proliferation, promoted apoptosis, increased the Bcl­2 associated X apoptosis regulator (Bax)/Bcl­2 apoptosis regulator (Bcl­2) ratio and caspase3/9 activity, and suppressed HLA­G protein expression in Hs 832(C).T cells. miR­148a downregulation using miR inhibitor significantly increased cell viability, inhibited apoptosis, and reduced the Bax/Bcl­2 ratio and caspase3/9 activity, and induced HLA­G protein expression in Hs 832(C).T cells. The GPER inhibitor, G15, suppressed GPER protein expression, upregulated miR­148a expression, decreased cell proliferation, promoted apoptosis, increased the Bax/Bcl­2 ratio and caspase3 activity, and suppressed HLA­G protein expression in Hs 832(C).T cells. The findings indicate that GPER/miR­148a/HLA­G signaling pathway may mediates the development of ovarian endometriosis and may become a potential therapeutic target for the treatment of endometriosis.

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.

[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.

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.