早孕绒毛滋养细胞FasL表达异常与自然流产的关系

通过比较正常早孕与自然流产者滋养细胞表面FasL表达,进一步从分子免疫学角度探讨自然流产的发病机理。方法：用特异性的FasL抗体进行免疫组化染色,并通过高清晰彩色病理免疫组化测量系统对其定量分析,SSPS对两组进行比较。结果：自然流产组滋养细胞表面FasL表达面积及强度均明显低于正常早孕组,两者间差异有显著性意义。结论：滋养细胞表面FasL表达减少,引起母胎间的免疫耐受的破坏,是导致自然流产的一重量免疫病因,诱导FasL的产生或调节母胎间的免疫耐受将为临床治疗自然流产提供新的方向。     

IL-10对早孕滋养细胞MMP-9表达的影响

人类滋养细胞的侵入仅限于子宫内膜及肌层的浅层 1/ 3,并于妊娠中期停止[1] 。这一过程受到多种因素的控制 ,ＩＬ 10在整个妊娠过程中都可由胎盘滋养细胞分泌 ,且α6整合素表达阳性的滋养细胞表面也存有ＩＬ 10的受体 ,故ＩＬ 10也是参与该调节的因子之一[2 ] 。侵入并非一个由细胞增殖造成的被动事件 ,而是一个主动的生化过程。滋养细胞通过分泌蛋白酶类获得侵袭能力 ,其中基质金属蛋白酶 9(ＭＭＰ 9)是参与滋养细胞侵入子宫内膜的主要蛋白酶[3 ] 。因此 ,为探讨ＩＬ 10对滋养细胞侵袭力的影响 ,我们采用ＲＴ ＰＣＲ方法 ,观察ＩＬ 10对早…     

研究滋养细胞侵袭的常用模型

正常妊娠绒毛滋养细胞(extravillous trophoblast,EⅥ)侵入子宫蜕膜、肌层内1/3及螺旋动脉,破坏螺旋动脉血管平滑肌、弹力纤维及神经组织,致螺旋动脉发生"血管重塑".目前,滋养细胞侵袭过程的调控机制是一个研究热点,现对滋养细胞侵袭研究的细胞模型及细胞外基质模型做一综述.     

人早孕滋养细胞与蜕膜基质细胞共培养对外周NK细胞的影响

为了分析人早孕母-胎界面功能细胞对外周NK细胞表型及功能的调节作用,我们模拟母-胎界面微环境,建立滋养细胞和蜕膜基质细胞共培养系统,并将此共培养体系与外周NK细胞共培养,采用流式细胞术分析NK细胞的表型和细胞因子的表达,用细胞毒检测试剂盒检测NK细胞的细胞毒活性。研究发现,母-胎界面滋养细胞与蜕膜基质细胞共培养体系能够显著上调外周NK细胞抑制型受体KIR2DL1的表达,下调外周NK细胞杀伤性受体NKp44的表达;显著上调Th2型细胞因子IL-10的表达水平,下调外周NK细胞穿孔素perforin表达及其细胞毒活性。结果表明,母-胎界面功能细胞能够诱导外周NK细胞的耐受表型及Th2型优势,下调外周及蜕膜NK细胞的细胞毒活性,从而有利于形成母-胎界面免疫耐受微环境。     

CXCR7分子对人早孕滋养细胞的调节作用

目的:已有研究表明CXCL12/CXCR4信号途径以自分泌的方式在人早孕滋养细胞增殖和信袭中发挥重要作用,探讨CXCL12的第二受体CXCR7信号对人早孕滋养细胞活力和侵袭功能的调节作用。方法:采用RT-PCR、免疫组化分析CXCR7在人早孕滋养细胞的表达;采用MTT和Transwell侵袭试验分别分析CXCL12/CXCR7信号途径对人早孕滋养细胞活力和侵袭功能的作用。结果:人早孕滋养细胞表达CXCR7分子,外源性给滋养细胞株HTR-8/Svneo添加anti-CXCR4(20μg/ml)和anti-CXCR7(10μg/ml)的中和性抗体后,均能显著抑制滋养细胞的增殖和侵袭能力。结论:人早孕滋养细胞表达CXCR7,通过CXCR7信号升调节滋养细胞的增殖和侵袭,其原因可能是与CXCR4形成二聚体共同参与CXCL12的信号传递。     

阿司匹林对TNF-α诱导人早孕滋养细胞增殖和侵袭的影响及机制

目的:采用体外细胞模型探讨阿司匹林促进人早孕滋养细胞增殖和侵袭在预防子痫前期(preeclampsia,PE)发生中的作用及机制。方法:收集PE病例20例和正常孕妇作为对照20例;HE染色观察胎盘组织病理改变;ELISA法检测基质金属蛋白酶2(MMP-2)及MMP-9的含量。体外培养人早孕滋养细胞HTR-8/SVneo,分别加入不同浓度的阿司匹林和肿瘤坏死因子α(TNF-α);CCK-8法检测细胞活力;Transwell实验检测细胞侵袭能力;Western blot检测细胞周期蛋白D1(cyclin D1)和增殖细胞核抗原(PCNA)的蛋白表达水平。结果:PE组胎盘组织存在子宫螺旋动脉血管重铸障碍,且孕妇血清中MMP-2及MMP-9浓度明显低于对照组(P0.05)。体外早孕滋养细胞系实验表明,与对照组比较,0.1 mmol/L阿司匹林明显增加细胞活力,且可缓解TNF-α引起的细胞活力降低(P0.05);TNF-α处理滋养细胞后,细胞cyclin D1及PCNA蛋白表达水平显著下降,侵袭能力减弱,培养上清液中MMP-2和MMP-9表达水平及活性均明显下降(P0.05);同时给予0.1 mmol/L阿司匹林和TNF-α处理后,滋养细胞cyclin D1及PCNA蛋白表达水平显著增高,侵袭能力增强,培养上清液中MMP-2和MMP-9表达水平及活性均明显上升(P0.05)。结论:小剂量阿司匹林可以促进绒毛滋养细胞增殖和侵袭,可能有利于改善PE胎盘浅着床和子宫螺旋动脉重铸异常情况。这为阿司匹林预防PE的发生提供实验依据。     

妊娠滋养细胞病变中FHIT和PTEN蛋白的表达

目的　探讨抑癌基因FHIT、PTEN蛋白表达在妊娠滋养细胞疾病发生、发展中的作用及临床病理意义。方法　采用免疫组化S P法分别检测了 30例完全性葡萄胎、15例侵袭性葡萄胎、15例绒毛膜细胞癌中FHIT、PTEN的蛋白表达。结果　完全性葡萄胎中FHIT蛋白表达低于正常早孕绒毛 (P <0 0 1) ,FHIT蛋白表达与葡萄胎滋养细胞增生呈负相关关系。侵袭性葡萄胎和绒毛膜细胞癌中FHIT蛋白表达均低于完全性葡萄胎 (P <0 0 5 ,P <0 0 1)。侵袭性葡萄胎和绒毛膜细胞癌中核PTEN蛋白表达均低于完全性葡萄胎 (P <0 0 1,P <0 0 1)。绒毛膜细胞癌中核PTEN蛋白表达与临床分期呈负相关关系。结论　FHIT和PTEN蛋白表达下降或缺失可能参与了妊娠滋养细胞疾病的发生、发展过程。     

Fas、Bcl-2在RAS胎盘滋养细胞中表达的相关性研究

目的探讨凋亡蛋白Fas、抑制凋亡蛋白Bcl-2在胎盘滋养细胞与RAS的相关性。方法选择20例RAS者作为研究组,同期正常妊娠20例作为对照组。采用免疫组织化学方法测定各组胎盘滋养细胞Fas、Bcl-2的表达。根据阳性表达率和表达强度进行量化评分,并进行相关性分析。结果(1)RAS组胎盘滋养细胞Fas、Bcl-2表达的免疫组化积分分别为4.61±0.57,1.69±0.53;对照组分别为1.344±0.61,4.79±0.48。其中Fas在RAS组的表达明显高于对照组(P<0.05),而Bcl-2则明显降低(P<0.05)。(2)Fas与Bcl-2在RAS组胎盘滋养细胞中的表达呈负相关(r=-0.63,P<0.05)。结论(1)胎盘滋养细胞Fas高表达、Bcl-2低表达可能与RAS的发生有关。(2)Fas与Bcl-2在胎盘滋养细胞中的表达失衡可能是RAS发生的原因之一。     

正常足月胎盘和胎儿宫内生长迟缓胎盘中滋养细胞的凋亡及相关基因表达

目的 比较正常足月胎盘与胎儿宫内生长迟缓胎盘（IUGR）中的滋养细胞凋亡指数,以及bcl-2和fas的表达,以探讨滋养细胞的凋亡与bcl-2和fas表达的关系。方法 取正常足月胎盘组织和I－UGR胎盘组织各5例,固定、石蜡包埋、切片,每组分别TUNEL法以及ABC法抗bcl－2和抗fas免疫组织化学染色。镜下计数滋养细胞的凋亡指数,并对bcl-2和fas的表达量进行显微图像分析。结果 与正常足月胎盘相比,IUGR胎盘的凋亡指数显著增多（P＜0．01）,bcl－2表达量显著减低（P＜0．01）,而fas表达量无显著差异（P＞0．05）。结论 IUGR胎盘滋养细胞的凋亡指数明显增多可能与细胞bcl-2表达量减少有关,而不受fas表达量的影响。     

早孕滋养细胞膜型及可溶性趋化因子CXCL16表达调控

目的 探讨趋化因子CXCL16在人早孕滋养细胞表达和释放的调控机制.方法 原代培养人早孕滋养细胞,实时定量RT-PCR、免疫化学和ELISA方法分析CXCL16的表达和分泌;ELISA方法分析细胞因子IFN-γ、TNF-α、IL-4刺激前后滋养细胞CXCL16的表达和分泌水平;ELISA方法分析ADAM10治疗前后滋养细胞CXCL16的表达及分泌水平.结果 滋养细胞表达并分泌趋化因子CXCL16;IFN-γ治疗后滋养细胞表达和分泌CXCL16水平均显著上升(P<0.01),但TNF-α和IL-4并不影响CXCL16的表达或分泌;ADAM10可以加速CXCL16自滋养细胞的脱落,但并不上调CXCL16的合成.结论 IFN-γ和ADAM10参与调控母胎界面滋养细胞趋化因子CXCL16的合成和分泌.     

抗人滋养细胞单克隆抗体的制备及其免疫生物学特性鉴定

研制抗人滋养细胞单克隆抗体并分析其免疫生物学特性。方法采用滋养细胞免疫Ｂａｌｂ／ｃ小鼠;用细胞ＥＬＩＳＡ法鉴定ｍＡｂ的特异性;用补体依赖的溶细胞作用测定ｍＡｂ的细胞毒性。结论利用细胞免疫法所获该组ｍＡｂ所针对的抗原表位,可能是诱发免疫性流产的滋养细胞膜特异性抗原。     

Diagnostic value of the plasmatic ADM level for early ectopic pregnancy

Objective: To analyze the plasmatic ADM level in early pregnancy and to investigate the diagnostic value of ADM in early ectopic pregnancy (EP). Methods: 70 patients with EP who had menopause for 5~8 weeks were included as study group, while 155 women with normal intrauterine pregnancy were also included as control group. The correlation between ADM level and menopause weeks was statistically analyzed and ROC curve was used to identify the diagnostic value of ADM. Results: (1) In 155 cases of normal intrauterine pregnancy, the plasmatic ADM level was increased with menopause weeks in linear relationship, and the correlation coefficient (R) was 0.991 (P<0.05). In 70 patients with EP, no significant increase was found with menopause weeks and no linear relationship can be found between ADM level and menopause weeks in EP group. The correlation coefficient (R) was 0.744 (P>0.05). (2) The multiple of median of plasmatic ADM level in EP group of menopause for 8 weeks was obviously lower than the intrauterine control group (P<0.01). (3) ROC curve was used to analyze the cut-off value of ADM level in the diagnosis of EP, and the area under the ROC curve was 0.523 (P>0.05) regardless of menopause weeks, however, the area under the ROC curve was 0.702 (P<0.05) at 8 weeks after menopause with sensitivity of 53.50% and specificity of 85.00%. Conclusions: Different from normal intrauterine pregnancy, plasmatic ADM level in early EP was relatively lower and no significant increase was found with menopause weeks; further studies are still needed for plasmatic ADM level as an indicator in the early diagnosis of EP.     

Chaperonin 10 in the rat oocytes and early embryos: its expression and activity for early pregnancy factor

PROBLEM: The aim of this study was to investigate the expression of chaperonin (cpn) 10 and cpn 60 mRNA in oocytes or embryos, and to further explore the possibility that early pregnancy factor (EPF) is identical with cpn 10. METHOD OF STUDY: The expressions of cpn 10 and cpn 60 mRNA in oocytes and embryos at the different stages (1-cell, 2-cell, 8-cell, and morula) were examined by polymerase chain reaction techniques. The EPF activity of native rat cpn 10 isolated from rat livers was evaluated by the rosette inhibition test. RESULTS: Similar levels of mRNA of cpn 10 and cpn 60 were detected in oocytes and embryos at every stage. There were no detectable EPF activities in the native cpn 10. Immunoprecipitation using polyclonal antibodies against cpn 10 did not affect the activity of EPF in the pregnant rat serum. CONCLUSION: Our results do not support the hypothesis that cpn 10 is identical with EPF.     

Endometrial expression of immunomodulatory cytokines and their regulators during early pregnancy in bonnet monkeys (Macaca radiata)

BACKGROUND: It is well established that endometrium undergoes extensive histological changes during implantation and subsequent stages of pregnancy in rodents as well as primates. Our previous investigation using a non-human primate model has demonstrated that morphological alterations are initiated even before the embryo invades the endometrium. The present study was undertaken to determine whether the embryo-induced morphological changes are accompanied by any alteration in the protein levels of the immunomodulatory cytokines and their regulators in the preimplantation stage endometrium. METHODS: The endometrial expression of immunosuppressive factors such as transforming growth factor beta2 (TGFbeta2), glycodelin (PP14), leukaemia inhibitory factor (LIF) and interleukin-6 (IL-6) were analysed on day 6 post-ovulation in pregnant and non-pregnant bonnet monkeys (Macaca radiata) using immunohistochemical methods. RESULTS: The endometrial expression of TGFbeta2, TGFbeta2 receptor, PP14 and IL-6 were significantly up-regulated (p < 0.05) in pregnant animals as compared to non-pregnant animals, whereas the expression of LIF and its receptor remained unaltered in pregnant animals. CONCLUSIONS: Expression levels of some immunomodulatory cytokines in endometrium are significantly increased even before the embryo invades the endometrium. The altered cytokine expression profile in endometrium probably contributes towards generating a conducive environment for the embryo survival, growth and development in the uterus.     

B超在早期宫角妊娠诊治中的临床应用价值

目的 探讨B超在早期宫角妊娠诊治中的临床应用价值.方法 选择川北医学院附属医院2005年6月至2010年6月临床拟诊早孕经B超诊断的早期官角妊娠自愿终止妊娠256例患者,年龄21～42岁.做二维或三维超声诊断,并在B超监测引导下施行人工流产或清官术,全部病例术后10d左右返院复诊.结果 全部病例经腹或经阴道B超(包括二...     

Predictive value of human chorionic gonadotrophin in the outcome of early pregnancy after in-vitro fertilization and spontaneous conception

This prospective study analyses the value of the -subunit ofhuman chorionic gonadotrophin (-HCG) in 120 pregnancies obtainedafter in-vitro fertilization (IVF)-embryo transfer. Spontaneousconception cycles (n = 16) were also analysed allowing a comparisonbetween these two forms of conception. Of the 120 clinical pregnancies,48 started as single gestations and 50 started with two or moresacs. There were 14 clinical abortions and eight ectopic pregnancies.All subjects had blood samples taken under a fixed protocolon days 11, 14, 17, 20 and 23 after follicular aspiration. Weeklysamples were obtained thereafter until day 60 from ovum retrieval.Transvaginal ultrasounds were performed at weekly intervals,starting on day 23 after follicular aspiration. In spontaneousconception cycles blood samples were obtained daily, startingon the day of follicular rupture. In spontaneous conceptioncycles and in IVF– embryo transfer conceptions, the doublingtime (DT) of ;-HCG was 1.4 ± 0.3 and 1.6 ± 0.4days respectively. This difference was not significant. In multigesta–tions,the DT was 1.5 ± 0.3 days. The absolute values of -HCGin early spontaneous gestations were significantly higher thanin IVF—embryo transfer cycles, suggesting that the blastocystimplants with less cellular mass when initiated in vitro ascompared with the in-vivo condition. The early prediction ofectopic pregnancy and spontaneous clinical abortion was analysedby the -HCG profile as well as the absolute values in comparisonto normal pregnancies. Both parameters showed significant differencesas early as the interval between days 11 and 23 from follicularaspiration. This study provides a comprehensive approach tothe evaluation of the outcome of early gestation in terms ofthe predictability of single and multigestation, ectopic pregnancyand early abortion.     

Study of early pregnancy factor (EPF) in equine (Equus caballus)

PROBLEM: Early pregnancy factor (EPF) is an immunosuppressive protein detected in the early pregnancy serum. We have already reported that we developed the rosette inhibition test for mare EPF and detected EPF in thoroughbreds. The aim of this study was to determine whether or not our method could be used clinically. METHODS OF STUDY: The rosette inhibition test for equine EPF was carried out on serum from six nonpregnant and six pregnant Shetland ponies, a female and a male Chinese pony, and four nonpregnant and 13 pregnant thoroughbred mares. In the thoroughbreds sera were collected during the pregnancy period. Furthermore, we measured progesterone and detected pregnant mare serum gonadotrophin (PMSG) in order to confirm pregnancy of the Chinese pony 3 and 6 months after mating. RESULTS: In the nonpregnant Shetland ponies, the rosette inhibition titre (RIT) was 6.0+/-1.0 and EPF was negative. In contrast, in the pregnant ponies, the RIT was 9.2+/-0.4 and EPF was positive. Based on these results, we diagnosed pregnancy of the Chinese pony. The RIT of the female Chinese pony (3 months after mating) was above 10 and EPF was positive. Furthermore, we detected PMSG and progesterone in the serum of this pony. EPF appeared in the maternal blood circulation at 24-72 hr after mating, it was detected until the second trimester, and after that it disappeared from the maternal serum. CONCLUSIONS: The pony's EPF was detected by using the same rosette inhibition test as in the thoroughbred and was present from 24 to 72 hr after mating until the second trimester. The results indicated that our method was useful for pregnancy diagnosis of Equine.     

Endocrinology: Outcome of inadvertent administration of a gonadotrophin-releasing hormone agonist (buserelin) in early pregnancy

All women undergoing pituitary down-regulation before plannedin-vitro fertilization (IVF) treatment in two IVF units werestudied to assess the risks of and to pregnancies occurringinadvertently when gonadotrophin-releasing hormone agonists(GnRHa) were used to achieve pituitary desensitization duringthe luteal phase prior to planned IVF treatment. In 2670 cycles,25 women conceived (0.9% of cycles). Of these, three resultedin pre-clinical abortions (12%) but there were no clinical abortions,and 22 have ended with live births at term of apparently normalinfants. Collation of these and other published data suggestthat pregnancy outcome is not adversely affected by GnRHa administrationin the luteal phase of the conception cycle.     

Bleeding patterns after vaginal misoprostol for treatment of early pregnancy failure

BACKGROUND: Dilatation and curettage (D&C) has been the usual treatment for early pregnancy failure (EPF). Medical management with misoprostol may be an effective alternative. Bleeding patterns during and after medical management of EPF are unknown. METHODS: A prospective cohort study was conducted at University-based clinics and physician offices. Eighty women <11 weeks estimated gestational age with a diagnosis of missed abortion or fetal demise were enrolled. Treatment consisted of either 800 micro g of moistened (2 ml of saline) or dry vaginal misoprostol. Self-reported bleeding and sanitary product usage were recorded in a daily 2 week diary. Haemoglobin was assessed at enrollment and 2 weeks later. RESULTS: After misoprostol treatment, patients reported bleeding or spotting every day for the 14 days observed. Self-assessed heavy bleeding days were few (median 3) and usually occurred immediately after treatment. Sanitary pad use was highly variable (mean 30.5, range 2-125 pads over the 2 week period) and not related to changes in haemoglobin. The mean decrease in haemoglobin was 0.5 g/dl (SD 1.2). Complete expulsion without D&C occurred in 85% of subjects. CONCLUSIONS: Bleeding for at least 2 weeks after vaginal misoprostol for EPF is common. Heavy bleeding is usually limited to a few days after treatment. Clinically important changes in haemoglobin are rare.