MUC16 对子宫内膜容受性的影响及其意义

目的:多囊卵巢综合征(Polycystic Ovary Syndrome,PCOS)系由于体内复杂的内分泌和代谢环境影响了子宫内膜稳态,导致子宫内膜容受性下降,造成患者生育力减弱和不良妊娠结局.通过测定多囊卵巢综合征患者子宫内膜与正常生育力妇女增生期、分泌早、中、晚期子宫内膜中MUC16的相对表达量,本文探讨MUC16与PCOS患者子宫内膜容受性下降的关系,为临床上改善PCOS患者子宫内膜容受性,提高PCOS患者的妊娠率,降低流产率提供一条新的可能途径.方法:选择PCOS患者子宫内膜、正常生育力妇女增生期、分泌早、中、晚期子宫内膜各20例,用免疫组化SP法检测MUC16在各组的表达情况.结果:(1)MUC16在月经周期各期均有表达,在分泌中期表达最强.(2)PCOS组MUC16的表达较分泌中期组弱,差异有显著性(P＜0.05).结论:(1)在子宫内膜中的表达呈周期性变化.(2)PCOS患者子宫内膜中MUC16表达异常可能使子宫内膜容受性下降,推测其与胚胎不能正常着床或着床后发育不良、导致流产有关.     

PCOS易感基因Hmga2在小鼠子宫蜕膜化中的表达与调节

目的研究PCOS易感基因Hmga2在子宫内膜容受性和蜕膜化中的表达与调节。方法通过早期妊娠、延期着床与激活、人工蜕膜化、卵巢类固醇激素处理等实验,利用q PCR、Western blot技术,阐述Hmga2在子宫内膜容受性中的作用。结果 Hmga2随着妊娠表达量逐渐增加,着床点与非着床点相比表达量显著升高,胚胎激活组比延迟着床表达量显著增高,人工诱导蜕膜化与非蜕膜化比较表达显著升高,Hmga2的表达与雌激素和孕激素呈正相关,体内受雌孕激素调节。结论表明Hmga2的表达与小鼠早期妊娠胚胎着床过程密切相关,参与子宫内膜蜕膜化过程,受活化胚泡和类固醇激素的影响。     

麒麟丸联合炔雌醇环丙孕酮对多囊卵巢综合征致不孕症患者子宫内膜容受性、血清性激素和氧化应激水平的影响

摘要 目的：探讨麒麟丸联合炔雌醇环丙孕酮对多囊卵巢综合征（PCOS）致不孕症患者子宫内膜容受性、血清性激素和氧化应激水平的影响。方法：选取我院于2018年1月~2020年1月期间收治的PCOS致不孕症患者460例,符合要求的患者按照随机数字表法分为对照组和研究组,各为230例。对照组患者予以炔雌醇环丙孕酮治疗,研究组予以麒麟丸联合炔雌醇环丙孕酮治疗,对比两组疗效、子宫内膜容受性、性激素、氧化应激、排卵率、妊娠率和不良反应。结果：研究组的临床总有效率较对照组更高（P<0.05）。研究组治疗后子宫动脉搏动指数（PI）、子宫动脉血流阻力指数（RI）低于对照组,子宫内膜厚度大于对照组（P<0.05）。研究组治疗后促卵泡激素（FSH）和黄体生成素（LH）低于对照组,雌二醇（E2）高于对照组（P<0.05）。研究组治疗后丙二醛（MDA）、活性氧物质（ROS）低于对照组,过氧化物歧化酶（SOD）高于对照组（P<0.05）。随访6个月后,研究组的妊娠率、排卵率明显高于对照组（P<0.05）。两组不良反应发生率组间对比无差异（P>0.05）。结论：麒麟丸联合炔雌醇环丙孕酮治疗PCOS致不孕症患者,可以有效改善患者子宫内膜容受性、血清性激素和氧化应激状态,提高妊娠率、排卵率,安全有效。     

钙网质蛋白在多囊卵巢综合征大鼠子宫内膜的表达及意义

目的：研究钙网质蛋白（CRT）在PCOS大鼠子宫内膜中的表达及生物学意义。方法：60只雌性SD大鼠随机分为PCOS组和对照组,每组各30只。给模型组24日龄大鼠皮下埋植左旋甲基炔诺酮硅胶棒3mm／只,3d后BID皮下注射人绒毛膜促性腺激素1．5IU。给对照组皮下注射等体积生理盐水。注射9d后观察大鼠卵巢形态学（HE染色）,化学发光法测定性激素水平。结果：模型组大鼠卵巢重量和体积均显著高于对照组（P〈0．01）。模型组大鼠卵巢出现类多囊卵巢综合征的改变。模型组卵巢各级发育期卵泡及黄体少见,卵泡多呈囊性扩张。模型组大鼠血清孕激素、睾酮、空腹胰岛素、空腹血糖水平均显著高于对照组（P〈0．05）;卵泡刺激素（FSH）水平显著低于对照组（P〈0．05）。LH／FSH比值显著高于对照组（P〈0．05）。采用免疫组织化学方法及灰度值测定,定量分析CRT在PCOS组和对照组的子宫内膜中表达。CRT在两组中的子宫内膜中均有表达。PCOS组子宫内膜上皮CRT表达显著低于对照组（P〈0．01）。结论：避孕硅胶棒联合hCG诱导SD大鼠多囊卵巢综合征模型是较好的PCOS模型。CRT与PCOS的发病密切相关．     

加味补中益气汤治疗肥胖型多囊卵巢综合征患者的疗效及对子宫内膜容受性和糖脂代谢的影响

摘要 目的：探讨加味补中益气汤治疗肥胖型多囊卵巢综合征（PCOS）患者的疗效及对子宫内膜容受性和糖脂代谢的影响。方法：选择2018年3月~2020年5月期间我院收治的肥胖型PCOS患者280例，采用抽签法分为观察组140例和对照组140例。对照组给予二甲双胍片治疗，观察组给予加味补中益气汤治疗，对比两组疗效、体质量指数（BMI）、腰臀比、性激素指标[雌二醇（E2），促黄体生成激素（LH）、卵泡刺激素（FSH）]、子宫内膜容受性指标[子宫内膜厚度、子宫内膜螺旋动脉搏动指数（PI）与阻力指数（RI）]、糖脂代谢指标[总胆固醇(TC)、低密度脂蛋白 (LDL-C)、甘油三酯(TG)、高密度脂蛋白 (HDL-C)、空腹血糖（FPG）]。结果：观察组的临床总有效率为92.14%（129/140），高于对照组的80.71%（113/140）（P<0.05）。治疗3个月经周期后，两组BMI、腰臀比、PI、RI、LH、FSH、TC、LDL-C、TG、FPG均下降，且观察组低于对照组（P<0.05）。治疗3个月经周期后，两组子宫内膜厚度、HDL-C、E2升高，且观察组高于对照组（P<0.05）。结论：加味补中益气汤治疗肥胖型PCOS患者，可降低患者体质量，调节性激素、子宫内膜容受性和糖脂代谢水平，具有较好的疗效。     

苍附导痰汤联合通元针法对脾虚痰湿型多囊卵巢综合征患者脂代谢、性激素和子宫内膜容受性的影响

摘要 目的：探讨苍附导痰汤联合通元针法对脾虚痰湿型多囊卵巢综合征（PCOS）患者脂代谢、性激素和子宫内膜容受性的影响。方法：选取2019年3月~2021年11月期间我院收治的PCOS患者87例，均辨证分型为脾虚痰湿。根据随机数字表法分为对照组（常规药物治疗，43例）和研究组（对照组基础上接受苍附导痰汤联合通元针法，44例）。对比两组临床疗效、中医证候积分、脂代谢指标、性激素指标和子宫内膜容受性变化情况。结果：研究组的临床总有效率为86.36%，高于对照组的62.79%，差异有统计学意义（P<0.05）。治疗后研究组中医证候积分较对照组低（P<0.05）。研究组治疗后的腰臀比（WHR）和体质量指数（BMI）、胰岛素抵抗指数（HOMA-IR）低于对照组（P<0.05）。研究组治疗后的卵巢体积小于对照组，内膜厚度厚于对照组（P<0.05）。研究组治疗后的黄体生成素（LH）、雌二醇（E₂）、卵泡刺激素（FSH）和抗苗勒管激素（AMH）低于对照组（P<0.05）。研究组治疗后的高密度脂蛋白胆固醇（HDL-C）高于对照组，总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）低于对照组（P<0.05）。结论：苍附导痰汤联合通元针法治疗脾虚痰湿型PCOS患者，可改善患者的脂代谢、性激素和子宫内膜容受性。     

GnRHR-Ⅱ在子宫内膜的分布及表达变化规律研究

目的:研究促性腺激素释放激素Ⅱ型受体(GnRHR-Ⅱ)在子宫内膜的分布及表达变化规律。方法:选择2015年1月~2016年7月期间我院收治的100例女性不孕患者,经诊断性刮宫技术获取50例增生期子宫内膜组织与50例分泌期子宫内膜组织,分别作为研究组与对照组;采用免疫组织化学染色法检测两组子宫内膜基质细胞与腺上皮GnRHR-Ⅱ的表达。结果:增生期、分泌期子宫内膜均有GnRHR-Ⅱ分布;研究组子宫内膜基质细胞的GnRHR-Ⅱ表达明显高于对照组,而腺上皮则明显低于对照组,差异有统计学意义(P0.05)。结论:GnRHR-Ⅱ在子宫内膜增生期与分泌期均有表达,且在膜基质细胞与腺上皮均有分布,其分布于表达变化规律在一定程度上与子宫内膜容受性有关,有望成为评估子宫内膜容受性的标记物。     

钙结合蛋白calbindin-D28k在人输卵管组织的表达

本研究旨在探讨育龄妇女输卵管中钙结合蛋白calbindin-D28k（CaBP-D28k）的表达和变化。33例月经规律、有正常生育史的育龄妇女,因子宫肌瘤、子宫腺肌症行子宫切除术（一并切除输卵管）,输卵管取材均包括峡部、壶腹部和伞部,按月经周期分为增生早期组6例,增生中期组5例、增生晚期组5例,分泌早期组7例,分泌中期组5例和分泌晚期组5例。采用免疫组织化学法和逆转录聚合酶链反应法检测CaBP-D28k在人输卵管组织中的表达。结果显示,人输卵管组织中存在CaBP—D28k蛋白及mRNA表达。CaBP-D28k蛋白表达于输卵管上皮细胞的胞浆中,在月经周期同一时期,输卵管峡部、壶腹部、伞部的表达无明显差别（P〉0．05）。在月经周期中,CaBP-D28k蛋白表达在增生早、中期最低,而在增生晚期和分泌早期明显增高（P〈0．05）,分泌中、晚期显著下降（P〈0．05）。CaBP-D28k蛋白阳性表达分布在月经周期中出现再分布,在增生早、中期呈灶状或片状弥散分布在胞浆中,增生晚期和分泌早、中期CaBP-D28k蛋白呈聚集的颗粒状集中在细胞顶部,至分泌晚期则仍呈灶状或弥散状分布;CaBP-D28k mRNA表达也随月经周期发生变化,在增生晚期和分泌早期明显增高（P〈0．05）。结果表明,人输卵管组织中存在CaBP-D28k蛋白及mRNA表达,且具有周期性变化。     

孕素1号引起的假孕家兔子宫内膜早熟

受精卵与子宫内膜发育的同步化是卵子着床的重要条件,这一原理已得到证实与公认。前文表明,复方孕素1号避孕药可引起人体子宫内膜提早转化:正常妇女子宫内膜一般于月经周期第16天由增生期转化为分泌早期,第21天发育到分泌晚期,第26天发育到行经前期;而服药对象内膜则早在周期第12、17和20天便发育到相应各期,较正常提早数日转化。作者推测,药物引起的子宫内膜提早转化,很可能干扰了内膜与卵龄的同步化,从而造成胚泡不能着床。并认为这可能是该药抗着床机制中的重要一环。为了探讨孕素1号的抗着床原理,验证上述设想,在家兔上建立了孕素1号抗着床模型。本工作即按照模型采用的给药时间与剂量,研究药物对假孕家兔引起的子宫内膜组织学变化,观察孕素1号是否同样可引起家兔内膜早熟?同时,利用卵子移植实验,观察早熟的内膜是否对移植卵的着床有影响,从而对孕素1号的抗着床原理提供一定的实验数据。     

补脾滋肾汤联合针灸治疗多囊卵巢综合征不孕症的临床疗效观察

摘要 目的：观察补脾滋肾汤联合针灸治疗多囊卵巢综合征（PCOS）不孕症的临床疗效。方法：选择2018年9月~2020年9月就诊的120例PCOS不孕症患者,随机分为观察组和对照组各60例。均给予口服枸橼酸氯米芬基础治疗,观察组在基础治疗上增加补脾滋肾汤联合针灸治疗,28 d为一个治疗周期,对比两组患者3个疗程后总有效率、中医证候量表评分、BMI（体质量指数）、月经情况、排卵情况、妊娠情况、血清性激素指标[T（睾酮）、LH（促黄体生成激素）、FSH（促卵泡生成激素）]、B超测定子宫内膜容受性指标[子宫内膜厚度、PI（子宫内膜螺旋动脉搏动指数）与RI（阻力指数）]。结果：治疗3个疗程后观察组的临床总有效率为90%（54/60）,高于对照组的83%（50/60）（P<0.05）。经过3个疗程治疗后,两组PCOS不孕症患者中医症候量表评分改善,月经情况改善,两组排卵率、妊娠率提高,且观察组高于对照组（P<0.05）。治疗3个疗程后,两组BMI、RI、PI、LH、T均下降,且观察组低于对照组（P<0.05）;两组子宫内膜厚度、FSH均增高,且观察组高于对照组（P<0.05）。结论：补脾滋肾汤联合针灸治疗多囊卵巢综合征不孕患者通过中药滋补脾肾、针灸治疗后,有效缓解了临床症状,性激素水平得到调节、体重减轻,提高调经促排卵助孕率,疗效确切。     

YKL-40和NF—KB在子宫内膜异位症中的表达及相关性研究

目的：通过检测人类软骨蛋白39（YKL-40）和核转录因子KappaB（NF—KB）在子宫内膜异位症组织中的表达,探讨二者与子宫内膜异位症的关系及二者的相关性。方法：用免疫组化二步法检测40例子宫内膜异位症（EMS）患者的异位内膜和在位内膜及40例子宫肌瘤患者子宫内膜（对照组）YKL-40和NF—KB的表达,并对检测结果进行统计学分析。结果：YKL-40、NF—KB在子宫内膜异位症患者的异位和在位内膜及对照组内膜中的表达率分别为67．5％、62．5％、35％及88．5％、57．5％、32．5％,差异有统计学意义（P〈0．01）;在不同月经周期YKL-40表达无统计学意义;在位内膜和正常内膜NF-KB的表达分泌期高于增殖期,差异有统计学意义（P〈0．05）;YKL-40和NF-KB在三种内膜中的表达具有正相关性,相关系数分别是0．305,0．267和0．457（P〈0．01）。结论：YKL-40和NF—KB在EMS发生发展中起重要的作用。     

Aquaporin-2 expression in human endometrium correlates with serum ovarian steroid hormones

The aim of the present study was to examine the expression of aquaporin-2 (AQP2), a member of the water channel family aquaporins (AQPs), in human uterine endometrium and its modulation of ovarian steroid hormone at the proliferative and secretory phases. Western blot, immunohistochemistry, and RT-PCR were employed in the present study. Western blot revealed a 29-kDa band that represented AQP2 in human endometrium. The expression of AQP2 in endometrium was confirmed by RT-PCR and immunohistochemical results. The immunohistochemical analysis demonstrated that AQP2 was prominent in luminal and glandular epithelial cells of endometrium. The levels of endometrial AQP2 expression changed during the menstrual cycle and were higher in the secretory endometrium than in the proliferative endometrium. A significantly high level of AQP2 was detected at the mid-secretory phase. There was a positive correlation between the levels of the endometrial AQP2 expression and the concentrations of the serum 17beta-estradiol (E2) or/and progesterone (P4). These data for the first time corroborate that AQP2 is expressed in human endometrium and that the expression of AQP2 in human endometrium might be regulated by E2 or/and P4. The changed expression of AQP2 at different phases of the menstrual cycle may be essential to reproductive physiology in human. The high level of endometrial AQP2 expression was observed at the mid-secretory phase, the time of embryo implantation, suggesting that AQP2 might play physiological roles in the uterine receptivity.     

The differences in RCAS1 and DFF45 endometrial expression between late proliferative, early secretory, and mid-secretory cycle phases

RCAS1 expression is related to the regulation of activated immune cells and to connective tissue remodeling within the endometrium. DFF45 seems to play an important role in the apoptotic process, most likely by acting through the regulation of DNA fragmentation. Its expression changes within the endometrium seem to be related to the resistance of endometrial cells to apoptosis. The aim of the present study was to evaluate RCAS1 and DFF45 endometrial expressions during ovulation and the implantation period. RCAS1 and DFF45 expression was assessed by the Western-blot method in endometrial tissue samples obtained from 20 patients. The tissue samples were classified according to the menstrual cycle phases in which they were collected, with a division into three phases: late proliferative, early secretory, and mid-secretory. The lowest level of RCAS1 and the highest level of DFF45 endometrial expression was found during the early secretory cycle phase. Statistically significantly higher RCAS1 and statistically significantly lower DFF45 endometrial expression was identified in the endometrium during the late proliferative as compared to the early secretory cycle phase. Moreover, statistically significantly higher RCAS1 and statistically significantly lower DFF45 expression was found in the endometrium during the mid-secretory as compared to the early secretory cycle phase. The preparation for implantation process in the endometrium is preceded by dynamic changes in endometrial ECM and results from the proper interaction between endometrial and immune cells. The course of this process is conditioned by the immunomodulating activity of endometrial cells and their resistance to immune-mediated apoptosis. These dynamic changes are closely related to RCAS1 and DFF45 expression alterations.     

Quantitative analysis of estrogen receptor mRNA in human endometrium throughout the menstrual cycle using a real-time reverse transcription-polymerase chain reaction assay

We conducted a quantitative analysis of ERalpha and ERbeta mRNA expression in normal human endometrium throughout the menstrual cycle in regular menstruating premenopausal women, taking advantage of this real-time PCR assay. Endometrial dating was determined from the histology of the endometrium and classified into: proliferative endometrium and secretory endometrium. Both ERalpha and ERbeta mRNA expression were detected in all endometrial samples at both proliferative and secretion phase. However ERalpha mRNA expression level was higher than that of ERbeta specially during proliferative phase. These results suggest that estrogenic effects occur predominantly through ERalpha than ERbeta.     

Concentration of plasminogen activators and inhibitor in the human endometrium at different phases of the menstrual cycle.

The concentrations of tissue plasminogen activator (t-PA), urokinase plasminogen activator (u-PA) and plasminogen activator inhibitor (PAI-1) have been determined in endometrial curettings obtained from 46 subfertile women during proliferative, early or late secretory phases of the menstrual cycle. t-PA activity and antigen concentrations was significantly higher (P < 0.001) in late secretory endometrium than in proliferative or early secretory endometrium. Higher concentrations of PAI-1 antigen (P < 0.05) were also noted in late secretory phase than in proliferative and early secretory endometrium. However, u-PA concentration was not significantly different and no PAI activity could be demonstrated in the menstrual phases studied. Zymography studies confirmed the presence of both t-PA and u-PA in the endometrium. Ovarian hormonal patterns may therefore influence the activity of plasminogen activators especially of t-PA in the endometrium during various phases of the menstrual cycle.     

The Human Endometrium Expresses the Glycoprotein Mucin-1 and Shows Positive Correlation for Thomsen-Friedenreich Epitope Expression and Galectin-1 Binding

Mucin 1 (MUC1) is a glycoprotein in human endometrium and is abundant at the luminal epithelial surface in the receptive phase. It has a highly glycosylated ecto-domain that contains keratan sulfate chains, that disappears at the time of implantation. In addition, the glycoforms on MUC1 differ in fertile and infertile women. Therefore the aims of this study were investigations on glycosylation of MUC1 with the Thomsen-Friedenreich (TF) epitope on normal human endometrium throughout the menstrual cycle and binding of galectin-1 on the TF epitope in the endometrium and the expression of galectin-1 on the human oocyte. Human endometrial tissue was obtained from 54 premenopausal patients and was immunohistochemically analyzed with monoclonal antibodies against MUC1, TF epitope, galectin-1, and biotinylated galectin-1. In addition, human oocytes were analyzed for TF, galectin-1 expression, and galectin-1 binding. We identified a significant upregulation of MUC1 and TF epitope and, in addition, galectin-1 binding in glandular epithelium and epithelial apical surface tissue from proliferative to secretory phase. With double staining experiments, we identified a coexpression of TF and MUC1 in the early secretory phase and galectin-1 binding to TF during the same period of time. In addition we identified TF epitope and galectin-1 expression plus binding on the human oocyte and irregularly fertilized oocytes. Upregulation of TF epitope on the glandular epithelium and epithelial apical surface tissue in the secretory phase and binding of galectin-1 at the same time show the possibility of galectin-1–mediated trophectoderm binding to the endometrium within the window of implantation. (J Histochem Cytochem 57:871–881, 2009)     

Characterization of periostin expression in human endometrium and endometriotic lesions

Objective(s): To investigate the expression of periostin in the eutopic and ectopic endometrium of women diagnosed as endometriosis and evaluate the role of periostin in the pathogenesis of endometriosis. Study design: In this study, the expression of periostin was evaluated in the endometrial specimens from 35 women diagnosed as endometriosis and from 30 healthy women. To assess the presence and localization of periostin throughout the menstrual cycle in both eutopic and ectopic endometrium of women with endometriosis, microscopic evaluation was conducted. It was also subsequently compared with normal endometrium. Results: In the eutopic and ectopic endometrium of women with endometriosis, immunoreactivities of periostin increased compared with those of normal endometrium. We also observed a cyclic variation in the eutopic stromal periostin immunoreactivity throughout their menstrual cycle because higher H score values were observed in the proliferative phase than those in the secretory phase. Conclusion(s): These findings indicated that periostin may be involved in the pathophysiology of endometriosis.     

Phenotypic analysis and proliferative responses of human endometrial granulated lymphocytes during the menstrual cycle

The in vivo function of the unusual population of CD56+ CD16- endometrial granulated lymphocytes (eGLs) in human endometrium is unknown; their increased numbers in the secretory phase of the menstrual cycle suggests that they may play a role in the immunobiology of nonpregnant endometrium. In the present study, the phenotype and proliferative responses of eGLs at various phases of the menstrual cycle were compared with those in early pregnancy. Endometrial GLs were highly purified (> 98% CD56+) using immunomagnetic separation, and the expression of cell surface antigens was examined in smears using a double immunohistochemical labeling technique. Proliferative responses to mitogens and interleukin 2 (IL-2) were assessed in hanging drops in 60-well Terasaki plates. There was low to no expression of CD3, CD8, CD16, HML-1, L-selectin, and CD25 (IL-2 receptor alpha) on CD56+ cells isolated from nonpregnant and pregnant endometrium. The expression of CD2, CD49a, and CD122 (IL-2 receptor beta, IL-2Rbeta), however, increased from the proliferative to the late secretory phase of the menstrual cycle. In contrast, CD11a, CD69, and CD49d expression was high and did not vary with menstrual cycle phase; CD49d levels were significantly reduced in early pregnancy. Unlike early-pregnancy eGLs, none of the CD56+ eGL cultures throughout the menstrual cycle displayed phytohaemagglutinin (PHA)-induced lymphoproliferation. In contrast, eGLs from nonpregnant endometrium in the presence of 5 or 100 U/ml IL-2 after 48- and 120-h incubation showed significant proliferative responses, as did eGL cultures from early pregnancy. A significantly reduced number of proliferative phase eGL cultures proliferated in response to IL-2 compared to secretory phase and early-pregnancy eGL cultures. The IL-2-induced proliferative responses of CD56+ eGLs were associated with increased IL-2Rbeta (CD122) expression. These findings demonstrate 1) differential eGL expression of CD2, CD49a, and CD122 during the menstrual cycle; 2) differential IL-2-induced eGL proliferative responses during the menstrual cycle; and 3) differences between eGLs from nonpregnant and pregnant endometrium in CD49d expression and their ability to respond to PHA.