二甲双胍对多囊卵巢综合征患者卵巢形态的影响

目的探讨对二甲双胍具有不同排卵反应的多囊性卵巢综合征（PCOS）患者的卵巢形态学变化。方法年轻、体重指数正常的PCOS患者接受6个月二甲双胍治疗中,选取20例治疗后仍无排卵者为无排卵组,20例有排卵但是未能成功妊娠者为排卵组。另选取20例年龄和体重指数与无排卵组、排卵组匹配的PCOS患者（未做任何治疗）作为对照组。检测3组患者的睾酮（T）、性激素结合球蛋白（SHBG）、卵泡刺激素（FSH）、黄体生成素（LH）、雌激素（E2）、催乳素（PRL）、空腹血糖、空腹胰岛素、动态平衡模型分析胰岛素抵抗指数（HOMA）、空腹血糖-胰岛素比值（G IR）,B超观察卵巢形态、体积的变化。结果二甲双胍治疗6个月后,无排卵组有3例（15%）、排卵组有17例（85%）出现卵巢形态学改变,对照组无一例出现卵巢形态变化。与治疗前比较,无排卵组治疗6个月后卵巢体积无明显变化[（12.2±1.2）cm3],而排卵组则明显下降[（11.5±1.6）cm3]。结论 PCOS患者使用二甲双胍治疗后可出现与排卵反应密切相关的卵巢形态学改变。     

少见部位异位妊娠31例临床分析

目的：探讨少见部位异位妊娠的临床特征及误诊原因。方法：对2001年2月～2011年2月我院收治的31例少见部位异位妊娠进行回顾性分析。结果：少见部位异位妊娠占同期异位妊娠的6.39%,误诊率为74.2%。除宫颈妊娠及宫角妊娠保守治疗成功者外,均需手术探查。结论：少见部位异位妊娠早期诊断较困难,对疑诊病例应及早清除病灶或手术探查,治疗效果好。     

稳定表达烟酸受体GPR109A的中国仓鼠卵巢细胞株的建立

目的 将GPR109A基因插入pEGFP-N3载体中构建重组质粒pEGFP-GPR109A,并建立稳定表达烟酸受体GPR109A的中国仓鼠卵巢(Chinese hamster ovary,CHO)细胞株.方法 构建重组质粒pEGFP-GPR109A,重组质粒转化大肠杆茵DH5a,重组质粒经PCR、酶切、测序验证正确后,应用脂质体转染技术将该质粒导入CHO细胞,用抗生素G418筛选稳定表达的细胞.倒置荧光显微镜观察克隆细胞株荧光信号,RT-PCR检测GPR109A基因mRNA表达,Westem Blotting检测绿色荧光蛋白与烟酸受体GPR109A的融合蛋白(GFP-GPR109A)的表达,激光共聚焦显微镜观察融合蛋白的细胞定位.结果 pEGFP-GPR109A真核表达质粒构建正确,融合蛋白GFP-GPR109A在CHO细胞中稳定表达,表达的融合蛋白主要定位于细胞膜.结论 成功构建pEGFP-GPR109A真核表达载体并建立其稳定表达的CHO细胞株;该细胞株的建立有助于GPR109A的更多生理病理功能研究,也为下一步抗动脉粥样硬化的药物筛选奠定了基础.     

卵巢成熟畸胎瘤患者术后发热相关因素研究

目的探讨卵巢成熟畸胎瘤患者术后发热相关因素。方法对2010年9月-2013年8月经手术治疗且病理确诊为卵巢成熟畸胎瘤患者进行研究,将88例术后发热患者纳入发热组,100例术后无发热的患者纳入对照组,对两组病历资料进行回顾性病例对照研究。结果发热组和对照组住院时间分别为（5.68±1.53）、（3.28±1.18）d,开腹手术构成比分别为38.64%（34/88）、20.00%（20/100）,瘤体最大径线分别为（6.65±3.07）、（5.33±1.87）cm,两组比较差异均有统计学意义（P〈0.05）;两组在年龄、体质量指数、术前体温、术前CA199、术前瘤体扭转、术前使用抗生素、瘤体特征（有无骨质、是否多房、是否双侧）、手术时间、术中出血量、盆腔粘连、安放腹腔引流管等方面差异均无统计学意义（P〉0.05）。结论卵巢成熟畸胎瘤患者术后发热与手术方式、瘤体大小相关,且影响住院时间;应当加强妇科检查,对于成熟畸胎瘤的早期发现和手术方式的合理选择均可减少术后发热的发生,从而缩短住院时间,节省医疗资源。     

Seasonal changes in plasma steroid levels in Indian major carp Labeo rohita: influence of homologous pituitary extract on steroid production and development of oocyte maturational competence

Circannual variations in plasma levels of testosterone (T), 17beta-estradiol (E(2)), and 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one (DHP) as well as seasonal fluctuations in ovarian steroid synthetic potential were observed in Indian major carp, Labeo rohita. A study was also conducted to examine the mechanism of the development of gonadotropin-induced maturational competence in oocytes of this fish. The present study recorded the lowest values of plasma E(2) and T in L. rohita during the period from October to January. A mild increase in the levels of these steroids observed in February was followed by their rapid rise reaching peak values in April, when the ovary contained mostly the vitellogenic follicles. In the month of May, as the postvitellogenic follicles predominated in the ovary, there was a decline in plasma concentrations of both T and E(2). Low levels of these steroids in plasma remained until January, except a small elevation detectable during June and July (spawning stage). DHP was not detected in the plasma of this fish collected during the period from August to March. Existence of DHP was first recorded in blood in the month of April (vitellogenic stage) and it quickly reached the peak value in May (postvitellogenic stage), followed by a sudden decline in the month of June. Under stimulation of fish pituitary extract (FPE), as a source of gonadotropin, in vitro production of E(2) and T by the vitellogenic follicles was shown to be highest compared to their production rate in other stages, while the postvitellogenic follicles recorded the highest rate of DHP synthesis. Acquisition of oocyte maturational competence (OMC) was shown to develop either by priming the vitellogenic stage fish with a single dose of FPE or by in vitro addition of FPE in culture. In vitro treatment of trilostane, an inhibitor of 3beta-HSD, blocked the FPE-stimulated steroid production but not the development of OMC. Presence of cycloheximide and actinomycin D in the incubation was shown to inhibit FPE-induced development of OMC, indicating the requirement of de novo protein synthesis for this process.     

Expression of P450 aromatase protein in developing and in sex-reversed gonads of the XX/XY type of the frog Rana rugosa

Gonadal differentiation in some species of amphibians is sensitive to steroids. The phenotypic sex of XX/XY-type frogs such as Rana rugosa can be reversed from female to male by injection of testosterone into tadpoles, but little is known about the molecular mechanism of this sex reversal. To elucidate the mechanism of the sex differentiation, we examined the role of P450 aromatase (P450arom), an enzyme that converts testosterone to estrogen, during gonadal differentiation of amphibians. In this study, we first cloned a P450arom cDNA homolog of the frog R. rugosa and analyzed by RT-PCR its expression profile in developing and in female-to-male sex-reversed gonads. P450arom expression was observed in the gonad of tadpoles during ovarian differentiation and became much stronger in the developing ovary in which only immature oocytes were observed. However, its expression declined significantly in the ovary of frogs 2 months after metamorphosis, when oocytes were growing; and it was no longer seen in adult ovaries. By RT-PCR, we also examined the expression of P450arom and SF-1 (steroidogenic factor-1; the orphan nuclear receptor) in the female-to-male sex-reversed gonad. The level of P450arom mRNA was high in the ovary, but it declined rapidly after the injection of testosterone. In contrast, no change in the SF-1 (also known as Ad4BP) expression was observed. Moreover, to identify the type(s) of cells expressing P450arom protein, we performed immunostaining with an antibody against frog P450arom protein. Cells giving positive signals were observed around oocytes in the ovary of frogs 1 month after metamorphosis. They were identified as follicle cells by both light and electron microscopy. The results, taken together, indicate that P450arom protein is synthesized in follicle cells and that P450arom is very much involved in ovarian differentiation in R. rugosa.     

Activation of protein kinase Czeta mediates luteinizing hormone- or forskolin-induced NGFI-B expression in preovulatory granulosa cells of rat ovary

We have previously demonstrated that luteinizing hormone (LH) induces a rapid and transient expression of NGFI-B in the ovary. In this report, we investigated the signaling pathway for LH- and forskolin-induced NGFI-B expression in cultured rat granulosa cells of preovulatory follicles. LH- or forskolin-induced NGFI-B expression was suppressed by high dose of protein kinase C (PKC) inhibitor RO 31-8220 (10 μM), but not by low doses RO 31-8220 (0.1–1.0 μM) or adenylate cyclase inhibitor MDL-12,300A, implicating the involvement of atypical PKCs. Kinase assay revealed that LH treatment of granulosa cells resulted in a rapid stimulation of atypical PKCζ activity. Interestingly, like LH, forskolin was also able to activate PKCζ. Treatment with the cell-permeable PKCζ-specific inhibitor pseudosubstrate peptide inhibited LH-or forskolin-induced NGFI-B expression, indicating the essential role of PKCζ. Consistent with this promise, in granulosa cells depleted of diacylglycerol sensitive PKCs by prolonged treatment with tetradecanoylphobol-13-acetate, LH or forskolin could still induce NGFI-B expression, and RO 31-8220 or the PKCζ pseudosubstrate peptide inhibited LH- or forskolin-induced NGFI-B expression. Furthermore, overexpression of dominant-negative PKCζ in primary granulosa cells using a replication-defective adenovirus vector resulted in the suppression of LH- or forskolin-induced NGFI-B expression. Our findings demonstrate that PKCζ, which is activated by LH or forskolin, contributes to the induction of NGFI-B in granulosa cells of preovulatory follicles.     

Tibolone: a compound with tissue specific inhibitory effects on sulfatase

While many transforming growth factor-β (TGFβ) superfamily ligands such as TGFβ, activin, and the bone morphogenic proteins (BMPs) are critical to the control of growth, differentiation, and cell fate, inhibin has a more limited role and is primarily responsible for the regulation of one hormone from one cell-type in the anterior pituitary. Inhibin is an endocrine hormone, produced by the gonads, that inhibits follicle stimulating hormone (FSH) release from the pituitary gonadotrope. The other hormones in the superfamily do not appear to act in an endocrine fashion, but rather control cell function in a paracrine or autocrine manner. Many components of the TGFβ/activin/BMP signal transduction pathway have been elegantly defined; however, the mechanism of inhibin action has not been completely dissected. Several cell surface proteins that associate with inhibin have been identified recently, and these molecules may provide the clues necessary to understand how inhibin regulates reproductive function.     

Involvement of calcium and calmodulin in the regulation of ovarian steroidogenesis in Atlantic croaker (Micropogonias undulatus) and modulation by Aroclor 1254

The involvement of calcium-dependent signal transduction pathways in the regulation of ovarian steroidogenesis was investigated in Atlantic croaker. Treatment with the calcium ionophores A23187 and ionomycin caused a 2- to 5-fold increase in basal steroid accumulation by croaker ovarian tissue in vitro. A23187 potentiated human chorionic gonadotropin (hCG)-induced testosterone (T) accumulation, whereas it inhibited accumulation of estradiol-17beta (E(2)) and the conversion of T to E(2), suggesting that intracellular calcium modulates aromatase enzyme activity. Gonadotropin stimulation of ovarian steroidogenesis was decreased in the presence of EGTA and inhibitors of voltage-sensitive calcium channels (VSCCs) and inositol-1,4,5-triphosphate-receptors (IP(3)Rs), indicating that releases of calcium from both intracellular and extracellular stores are components of the signal transduction pathways initiated by gonadotropin. Calmodulin is also involved in the regulation of ovarian steroidogenesis in croaker, since the calmodulin inhibitors W-7 and trifluoperazine (TFP) attenuated hCG-stimulated T and E(2) accumulation. These results are broadly similar to those reported previously in goldfish and suggest that the major calcium-dependent signaling pathways involved in gonadotropin stimulation of ovarian steroidogenesis in tetrapods are also present in teleosts. In addition, the involvement of calcium in the regulation of aromatase activity was demonstrated for the first time in a vertebrate ovary. Finally, acute exposure to 0.001-1 ppm Aroclor 1254 induced up to a 5-fold increase in hCG-stimulated E(2) accumulation, and this effect was attenuated by co-treatment with inhibitors of VSCCs and calmodulin, suggesting the existence of a novel mechanism of endocrine disruption by an environmental contaminant involving alteration of calcium-dependent signaling pathways regulating steroidogenesis.