肿瘤转移抑制因子CD63/ME491基因在小鼠子宫内膜表达的动态观察

目的 了解小鼠动情周期和早孕子宫中CD63/ME491 mRNA和蛋白的表达规律,探讨CD63/ME491在胚胎着床过程中的作用.方法 应用RT-PCR和免疫组化技术分别观察CD63/ME491 mRNA和蛋白的表达.结果 在整个动情周期中,CD63/ME491 mRNA在动情间期表达最多,而在动情期表达最少,但蛋白质却是在动情前期和间期表达最丰富,子宫内膜上皮和基质细胞均呈阳性.早孕的小鼠子宫组织均有CD63/ME491 mRNA表达,且在胚胎开始植入的第4天表达最多,以后维持在较高的表达水平.CD63/ME491蛋白在妊娠第1～6天子宫内膜腔上皮细胞和腺上皮细胞呈阳性表达.但在基质细胞表达的量和范围却不同:妊娠第1天,无CD63/ME491蛋白的表达;妊娠第2天,子宫内膜基质细胞出现微弱阳性表达;以后CD63/ME491蛋白在基质细胞的表达量和表达范围逐渐增强.结论 在胚胎着床过程中,CD63/ME491 在小鼠子宫中呈动态表达,提示它可能参与了子宫内膜对滋养层细胞有限侵袭的调控.     

瘦素对小鼠着床前胚胎发育影响的体外研究

目的　探讨瘦素在体外对小鼠着床前胚胎发育的影响。方法　 1收集小鼠 2 -细胞胚胎 ,在不同剂量瘦素的 CZB培养液中进行体外培养 ,观察胚胎发育情况 ,并进行胚胎移植 ,观察着床率 ;2收集小鼠 2 -细胞胚胎在空白 CZB培养液中体外培养至桑葚胚期 ,再分别置入不同剂量瘦素的 CZB培养液中进行体外培养 ,观察胚胎进一步发育情况。结果　瘦素能提高 2 -细胞胚胎体外发育的质量、发育率和胚胎着床率。当瘦素剂量为 10 ng/ ml时平均胚胎形态学评分 (AES)为 16 .0 8± 0 .37,剂量为 5 0 ng/ ml时 AES为 17.5 7± 0 .4 2 ,两者间差别有统计学意义(P<0 .0 5 )。但剂量进一步增加 ,促进作用不再递增。瘦素对桑葚胚体外进一步发育无显著影响。结论　瘦素参与了小鼠着床前胚胎的发育 ,能提高胚胎发育质量、发育率 ,有利于胚胎的进一步着床。     

肿瘤转移抑制因子CD82/KAI1基因在小鼠子宫内膜的动态表达

0 引言 原癌基因/癌基因的激活和/或抑癌基因(包括肿瘤侵袭转移抑制基因)的失活、基因突变等是胚胎植入这一正常生理过程信号转导通路所必需的[1]. 本实验我们观察小鼠动情周期和妊娠早期,CD82/KAI1 mRNA和蛋白在子宫组织中的表达情况,探讨它们在胚胎植入信号转导通路中的作用.     

针刺对胚胎着床障碍大鼠子宫CD68+巨噬细胞的影响

目的 观察针刺对胚胎着床障碍大鼠子宫CD68+ 巨噬细胞的影响。方法 将48只妊娠大鼠随机分为正常组(N)、模型组(M)、针刺组(A)和黄体酮组(P)。其中M、A 和P组于妊娠第1天(D1)给予米非司酮-麻油溶液造模,N 组作为对照仅给予等量麻油溶液。同时,A 组用自制布袋固定并于D1开始每天针刺三阴交、后三里,N 组和M 组仅固定,P组则肌肉注射黄体酮,直至处死。于D8、D10分别收集各组子宫样本,采用免疫组织化学法、Westernblot和Real-timePCR等方法测定子宫CD68的表达。结果 与N 组相比,M 组大鼠D8、D10子宫CD68蛋白和mRNA 的表达均明显降低(P <0.05);与M 组相比,A 组和P组大鼠D8、D10子宫CD68蛋白的表达明显增高(P<0.05);与M 组相比,A 组大鼠D8和P组大鼠D8、D10子宫CD68mRNA 的表达明显增高(P<0.05)。结论 针刺可能通过调节大鼠子宫CD68+ 巨噬细胞的表达而改善胚胎着床障碍,从而促进胚胎着床。     

小鼠子宫内膜胚泡着床点及旁组织白血病抑制因子基因表达研究

目的:探讨白血病抑制因子(LIF)基因在小鼠胚泡着床窗口期了宫内膜表达的特定部位.方法:采用半定量逆转录聚合酶链反应(RT-PCR),对20只孕期第4天的小鼠子宫内膜着床点和旁组织的LIF mRNA进行检测.结果:20只孕期第4天的小鼠着床点LIF mRNA表达平均水平为0.5316,旁组织LIF mRNA表达平均水平为0.3711.统计处理t检验差异具有显著性(P＜0.05).结论:在胚泡着床窗口期,小鼠子宫内膜胚泡着床点和旁组织LIF基因表达存在差异.     

白血病抑制因子调节胚胎发育与着床的研究进展

白血病抑制因子(Leukemia inhibitory factor,LIF)是参与调节胚胎着床过程的重要的细胞因子之一,尽管如此,LIF及其受体在不同种属动物中的表达又不完全相同,孕酮和雌激素对其分泌的调节作用在不同种属动物之间、体内外之间,甚至于同种动物也都不完全一样。LIF调节着床机理的研究正日益受到广泛的重视。     

早孕小鼠胚胎着床窗口期子宫内膜mmu-miR-106b表达规律及其作用

目的：探讨mmu-miR-106b在早孕小鼠胚胎着床过程中的表达规律,阐明其对胚胎着床的调控作用。方法：应用real-time PCR和原位杂交检测mmu-miR-106b在早孕小鼠孕4 d、孕5 d及孕6 d子宫内膜中的表达;子宫内膜基质细胞转染mmu-miR-106b的模拟物和抑制剂后,MTT和流式细胞仪检测细胞的增殖凋亡情况;结合靶基因预测数据库,利用Western blot确定mmu-miR-106b在子宫内膜中的靶基因。结果：mmu-miR-106b在小鼠孕6 d子宫内膜的表达较孕4 d明显下调（P=0.039）,孕5 d着床点与着床旁组织表达无明显差异（P=0.606）,定位于子宫内膜基质细胞;上调mmu-miR-106b会促进子宫内膜基质细胞的增殖,下调其表达会促进细胞凋亡;通过miRGen、Targetscan、Pictar数据库筛查获得与胚胎着床相关的靶基因核磷蛋白1、肿瘤易感基因１０１（tumor susceptibility gene 101,Tsg101）和10号染色体缺失的同源性磷酸酶张力蛋白等,其中Tsg101的表达受到mmu-miR-106b负调控（P=0.042）。结论：mmu-miR-106b可能通过靶向Tsg101,影响胚胎着床过程中子宫内膜基质细胞增殖,对胚胎着床发挥调控作用。     

LIF对体外培养小鼠孕早期胚胎发育及ICAM-1表达的影响

目的研究白血病抑制因子（ＬＩＦ）在胚胎发育中的作用并探讨其参与胚泡着床的机制。方法收集昆明种小鼠的单细胞受精卵,连续培养１２０ｈ,观察不同浓度ＬＩＦ对胚胎发育的影响;用免疫印迹法测定培养液中细胞间粘附分子（ＩＣＡＭ－１的表达水平。结果ＬＩＦ在０．１～１０ｎｇ／ｍｌ浓度之间对胚胎发育有促进作用,其作用在桑椹胚及胚泡期最明显（Ｐ＜０．０５）;ＬＩＦ增加胚胎ＩＣＡＭ－１的表达,其作用在０．１ｎｇ／ｍｌ浓度时最强,并随着浓度的增加而减弱。结论适应浓度的ＬＩＦ可促进胚胎发育,增加ＩＣＡＭ－１的表达,从而参与胚胎着床过程。     

应用组织芯片技术检测结直肠癌组织中KAI1/CD82和ME491/CD63的表达

目的 研究结直肠癌组织中KAI1/CD82和ME491/CD63的表达水平及其临床病理意义.方法 应用组织芯片和免疫组织化学技术检测232例结直肠癌组织中KAI1/CD82和ME491/CD63的表达,并运用统计学方法分析KAI1/CD82和ME491/CD63的表达与结直肠癌各种临床病理特征的关系及两者的相关性.结果 KAI1/CD82在结直肠癌组织中的表达与肿瘤的分化程度和Dukes分期有关(P＜0.05),与患者的性别、年龄、浸润深度、淋巴结转移无关(P＞0.05).未发现ME491/CD63表达与结直肠癌病理资料之间差异有统计学意义(P＞0.05).KAI1/CD82和ME491/CD63的表达差异有统计学意义(P＜0.05).结论 KAI1/CD82在结直肠癌组织中的表达与肿瘤临床病理特征密切相关,可作为结直肠癌的预后因子.KAI1/CD82和ME491/CD63的表达相关.     

Effect of Bushen Yiqi Hexue recipe on pinopodes expression on endometrial surface in embryo implantation dysfunctional mice

Objective: To explore the therapeutic mechanism of Bushen Yiqi Hexue recipe (BYHR), and observe its effect on pinopodes expression on endometrial surface in mifepristone induced embryo implantation dysfunctional mice.Methods: Pregnant Kunming mice were randomly divided into the normal group, the control group and the treated group. Mice in the treated group were administered with BYHR, and those in the normal group and the control group were administered with normal saline, starting from the very first day of pregnancy (Pd1). On the Pd4, mifepristone (RU486) was subcutaneously injected into mice in the control group and the treated group. Pinopodes expression on endometrial surface at 09:30–10:00 pm on Pd4 (regarded as time point 1, T1) and at 09:30–10:00 am on Pd5 (as time point 2, T2) was determined by scanning electron microscopy. Pregnancy rate and average implanted embryos were observed on Pd7.Results: Pregnancy rate and average implanted embryos in the control group were obviously lower than those in the normal group (allP<0.01), while the two parameters were significantly higher in the treated group (P<0.05,P<0.01, respectively) than those in the control group. In the normal group, abundant developing pinopodes were distributed over the whole endometrial surface in T1, and they were altered to a great deal of fully developed pinopodes in T2. But in the control group, only a few pinopodes were expressed locally on endometrial surface in T1, showing a nonsynchronous figure of development, and they completely disappeared in T2. In the treated group, a lot of developing pinopodes were expressed like those in the normal group but somewhat lagged behind, whereas many fully developed pinopodes were expressed in T2.Conclusion: The decrease of the pinopodes in Tl and the cleaning up of them in T2 is possibly the partial mechanism of mifepristone in inducing embryo implantation dysfunction. It is indicated that BYHR could improve pinopodes expression on endometrial surface, eventually better the uterine receptivity and improve the embryo implantation. Project supported by the National Natural Science Foundation of China (No. 30171193)     

Down-regulation of coxsakie and adenovirus receptor during embryo implantation

In this study,real-time PCR and immunohistochemistry were used to detect coxsakie and adenovirus receptor (CAR) expression.Both localization and quantity were evaluated in the uteri ob-tained at days post coitus (dpc) 2.5,4.5,6.5,8.5.Outcome of PCR was assessed by 2-ΔΔCt method.Im-age Pro-Plus 6.0 software was used for quantifying mean density of CAR expression in immunohisto-chemical sections.We found relatively weak CAR expression in the mouse uteri during implantation window.PCR and immunohistochemistry revealed highest CAR expression was detected on dpc 2.5 followed by down-regulation of CAR at dpc 4.5 and 6.5 (with significant difference).At dpc 8.5,CAR expression was increased slightly again.It is concluded that during implantation,the expression of CAR mRNA and protein is declined,resulting in the impairment of tight junction between cavity epithelium cells.After implantation window closure,CAR appears again to maintain epithelium stability.CAR might play an important role during embryo implantation procedure.     

Effect of Acupuncture on CXCL8 Receptors in Rats Suffering from Embryo Implantation Failure简

To observe the effect of acupuncture on CXCL8 receptors（CXCR1 and CXCR2） in rat endometrium experiencing embryo implantation failure, 72 pregnant rats were randomly divided into four groups： normal group（N）, embryo implantation failure group（M）, acupuncture treatment group（A）, and progestin treatment group（W）. Then the rats in each group were equally randomized into a day-6（D6） group, a day-8（D8） group, and a day-10（D10） group. The rats in group M, group A, and group W were treated with mifepristone-sesame oil solution on day 1, while the rats in group N were injected with the same amount of sesame oil. Meanwhile, ＂Housanli＂ and ＂Sanyinjiao＂ were selected for acupuncture. From day 1 to the time of death, the rats in group A were fastened up and then acupuncture was administered while the rats in group N and group M were only fixed, and the rats in group W were given progestin. The number of implanted embryos was calculated. The expression of CXCR1 and CXCR2 in rat endometrium was detected by immunohistochemistry, Western blotting and real-time PCR. Compared to group N, the average number of implanted embryos, the protein and mRNA expression of CXCR1（D6, D8 and D10）, and the protein and mRNA expression of CXCR2（D8 and D10） in rat endometrium were significantly decreased in group M. Compared to group M, there was significant elevation in the average number of implanted embryos, the protein expression（D6, D8 and D10） and mRNA expression（D8） of CXCR1 in rat endometrium of group A, and the protein expression（D8 and D10） and mRNA expression（D8） of CXCR2 in rat endometrium of group W. These findings indicated that acupuncture can increase the number of implanted embryos in rats of embryo implantation failure, which may be relevant with up-regulation the expression of CXCR1 and CXCR2 at maternal-fetal interface of rats with embryo implantation failure.     

巨噬细胞移动抑制因子在围植入期小鼠子宫内膜中的表达

目的探讨巨噬细胞移动抑制因子(MIF)在围植入期小鼠子宫内膜中的表达分布状况及其意义。方法应用免疫组化SP法检测妊娠1～8d小鼠子宫内膜MIF的表达情况,并以动情期非妊娠小鼠子宫做对照。结果在小鼠子宫内膜的所有组织均可检测到MIF的表达,主要表达于腺上皮、腔上皮及基质中的细胞团。MIF在妊娠1～3d表达较强,妊娠4～5d表达较弱,主要表达于子宫内膜的腔上皮和腺上皮;妊娠6～8d,MIF表达再次增强,但主要表达于基蜕膜和次级蜕膜区。结论妊娠1～8d小鼠子宫持续表达MIF,提示MIF可能参与小鼠胚泡植入及胚泡植入后的发育过程。     

苯妥英钠对动脉粥样硬化性脑梗死病人HDL-c、CD62p/CD63的影响

目的:观察苯妥英钠对动脉粥样硬化性脑梗死病人HDL-c、CD62p/CD63的影响,为动脉粥样硬化性脑血管病的预防提供理论依据。方法:选择门诊或住院动脉粥样硬化的脑血管病病人32例作为治疗组,以苯妥英钠0.2g/d进行治疗,另选择32例相似病例作为对照组,两组病人除苯妥英钠治疗外,其余治疗均相同,治疗前及治疗后30d、90d分别抽取晨空腹血检查HDL-c、CD62p/CD63水平,统计分析治疗前后HDL-c、CD62p/CD63变化百分比评价疗效。结果:治疗组病人在治疗后30d,血清HDL-c水平即开始升高,90d后升高更明显(P<0.01),两组病人血清CD62p阳性的血小板比例在治疗后30d即见明显减少,与治疗前相比均有极显著差异(P<0.01),治疗组与对照组比较差异不明显(P>0.05),治疗90d后进一步减少,治疗90d后治疗组CD62p阳性的血小板比例较对照组下降幅度更明显(P<0.05);治疗组CD63阳性的血小板比例呈逐渐下降趋势,而对照组仅在病后90d才略有下降,治疗组在治疗后90dCD63阳性的血小板比例下降幅度明显大于对照组(P<0.001)。结论:苯妥英钠对血清HDL-c水平有明显的提升作用,对急性脑梗死后血清CD62p及CD63阳性的血小板比例的改变有一定的调节作用。     

血必净注射液对脓毒症患者血小板活化因子CD62P和CD63的影响

目的 探讨血必净注射液对脓毒症患者血小板活化水平的影响。方法 选取2019年6月—2020年1月潍坊医学院附属医院抢救监护室（EICU）、重症监护治疗病房（ICU）收治的脓毒症患者42例进行前瞻性队列研究。通过随机分配原则将其分为常规治疗组与血必净组。收集患者入院24 h的基础资料及临床数据，同时比较两组的基础资料、CD62P和CD63的阳性表达率、序贯性器官功能衰竭评分（SOFA评分）和急性生理与慢性健康状况评分（APACHE Ⅱ评分）；采用Spearman法分析CD62P和CD63与凝血相关指标、SOFA评分及APACHE Ⅱ评分之间的相关性；采用多因素Logistic回归分析脓毒症患者14 d全因死亡的独立危险因素。结果 两组患者的年龄、性别、高血压、糖尿病、肾病、心脑血管病、感染部位比较，差异无统计学意义（P >0.05）。常规治疗组与血必净组治疗第8天临床资料比较，血必净组血小板计数水平升高（P <0.05），降钙素原（PCT）水平、中性粒细胞百分比（NEUT%）水平、纤维蛋白原（FIB）水平、凝血酶原时间（PT）水平、部分活化凝血酶时间（APTT）水平、APACHE Ⅱ评分、SOFA评分、病死率、住院时间、CD62P阳性表达率、CD63阳性表达率下降（P <0.05）。Spearman相关性分析结果显示，血小板活化因子CD62P和CD63的阳性表达率与血小板计数水平呈负相关（P <0.05），CD62P和CD63阳性表达与PT水平、APTT水平、SOFA评分、APACHE Ⅱ评分呈正相关（P <0.05）。多因素Logistic回归分析发现，CD62P和CD63阳性表达是脓毒症患者14 d死亡的独立危险因素（P <0.05），血小板计数水平是其独立保护因素（P <0.05）。结论 血必净注射液可能通过抑制脓毒症患者血小板活化因子CD62P、CD63的阳性表达率，降低血小板活化水平，从而降低14 d全因死亡率，改善脓毒症患者的近期预后。     

甲氧滴滴涕染毒对小鼠着床期HOXA-10基因表达的影响

目的：观察甲氧滴滴涕慢性染毒对雌鼠HOXA-10基因表达的影响．方法：使用5wk龄雌性昆明小鼠作为实验动物,按体质量随机分为4组,每组10只,经2wk适应性饲养后腹腔注射染毒,染毒剂量分别为0（对照）、16（低剂量）、32（中剂量）、64（高剂量）mg/（kg·d）,对照组给予等剂量芝麻油,连续15d后合笼,继续给药4d,于孕5d处死．结果：①染毒后小鼠一般情况未见明显异常;②免疫荧光显示：对照组荧光明显亮于染毒组;③实时荧光PCR测得的HOXA-10 mRNA表达：与对照组比较,中、高剂量组均显著降低HOXA-10 mRNA的表达水平（P〈0．05）．结论：MXC染毒可降低孕鼠子宫组织中HOXA-10基因的表达从而影响胚胎着床．