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。     

胚胎着床的子宫内膜免疫学研究现状与展望

胚胎着床的过程极为复杂,包括定位、黏附和侵入三个阶段.着床仅发生在一个极其有限的时间与空间范围内(即所谓的"着床窗"),需要子宫内膜和胚胎准确的同步化、相互作用和配合.     

S100A2在围着床期小鼠子宫内膜表达的动态观察

目的:了解S100A2 mRNA及蛋白在围着床期小鼠子宫的表达情况,为阐明S100A2在胚胎着床过程中的作用积累实验依据。方法:应用荧光定量PCR(FQ-PCR)和免疫组化技术分别检测S100A2 mRNA和蛋白的表达情况。结果:①S100A2mRNA在妊娠第1～6天子宫中均有表达,且表达量逐渐降低;同时,在第5天,着床点的含量明显低于非着床点。②S100A2蛋白主要表达于小鼠子宫内膜基质细胞的胞浆中,其表达量在第1、2天很丰富,第3、4、5天很低。③子宫内膜的腔上皮和腺上皮细胞未见S100A2表达。结论:在胚胎着床过程中,S100A2在小鼠子宫中的表达呈动态变化,提示可能参与了子宫内膜对滋养层细胞侵袭的调控。     

S100A2在围着床期小鼠子宫内膜表达的动态观察

目的：了解S100A2 mRNA及蛋白在围着床期小鼠子宫的表达情况,为阐明S100A2在胚胎着床过程中的作用积累实验依据。方法：应用荧光定量PCR（FQ-PCR）和免疫组化技术分别检测S100A2 mRNA和蛋白的表达情况。结果：①S100A2 mRNA在妊娠第1～6天子宫中均有表达,且表达量逐渐降低;同时,在第5天,着床点的含量明显低于非着床点。②S100A2蛋白主要表达于小鼠子宫内膜基质细胞的胞浆中,其表达量在第1、2天很丰富,第3、4、5天很低。③子宫内膜的腔上皮和腺上皮细胞未见S100A2表达。结论：在胚胎着床过程中,S100A2在小鼠子宫中的表达呈动态变化,提示可能参与了子宫内膜对滋养层细胞侵袭的调控。     

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。     

子宫内膜离子通道对子宫内膜容受性的影响

子宫内膜容受性是自然妊娠和辅助生殖技术成功妊娠的关键因素之一.子宫内膜离子通道是调节子宫内膜接受性和胚胎植入的重要方法.子宫内膜接受性是指在一定的时间内,胚囊能够定位、黏附并穿透子宫内膜,从而引起子宫内膜间质的一系列变化,即胚胎植入的能力.离子通道的异常表达可导致子宫内膜接受损伤和(或)植入失败.进一步研究探讨子宫内膜...     

二补助育汤对胚胎着床障碍模型小鼠子宫内膜自噬基因 Beclin-1、LC3 B表达的影响

目的探讨二补助育汤对胚胎着床障碍模型小鼠胚胎着床率及子宫内膜自噬基因Beclin-1、微管相关蛋白轻链3(LC3B)mRNA及蛋白表达的影响。方法将50只妊娠ICR雌性小鼠随机分为空白组、模型组、二补助育汤低剂量组、二补助育汤高剂量组、阿司匹林组,每组10只。用米非司酮造成胚胎着床障碍模型,每组每日以相应药物灌胃,于妊娠第5天下午脱颈处死,观察各组子宫形态,计算各组妊娠率、平均着床位点数,检测子宫内膜Beclin-1、LC3B mRNA及蛋白的表达量。结果空白组小鼠子宫色泽红润,胚胎着床部位膨大,分布均匀;模型组小鼠子宫色白、较细;用药各组子宫较细,色泽淡红。模型组平均着床位点数较空白组减少(P0.001);与模型组相比,二补助育汤高剂量组与阿司匹林组平均胚胎着床位点数均增多(P0.05)。模型组小鼠子宫内膜Beclin-1、LC3B mRNA及蛋白表达水平均低于空白组(P0.05);与模型组相比,二补助育汤低、高剂量组与阿司匹林组Beclin-1、LC3B mRNA及蛋白表达水平均增加(P0.05)。结论二补助育汤可增加Beclin-1、LC3B mRNA及蛋白的表达,提高子宫内膜细胞自噬水平,从而改善子宫内膜容受性增加平均着床位点数。     

雌三醇对大鼠的抗着床作用及其对卵巢黄体和子宫内膜的影响

The anti-implantation effect of estriol on rats was studied by enzyme histochemical methods. The activities of 3beta-hydroxy-steroid dehydrogenase (3beta-HSD), AcP in corpus luteum, A1P in endometrium of treatment groups were compared with those of control groups. In order to know the effect of estriol on endometrium during preparing for blastocyst implanting, we counted the number of mitosis cell in endometrium of 4th day pregnant rat under microscope. Serum progesterone level of early pregnant rats was measured by radioimmunoassay the results were as follows: 1. Estriol administrated postcoitally could prevent the early pregnancy of rats by the way of anti-implantation of blastocyst; 2. Activity of ovarian corpus luteal 3beta-HSD was depressed, whereas activity of AcP increased in the treatment group; 3. The number of mitotic cell in endometrial stroma was: in the treatment group 0.2 +/- 0.2, in the control group 45.7 +/- 16.2 (P less than 0.001); A1P reaction of endometrial stroma was negative in the treatment group, whereas it was positive in the control group; and 4. The serum progesterone level was: in the treatment group 17.4 +/- 12.6, in the control group 39.3 +/- 11.1 ng/ml (P less than 0.01). The results suggest that estriol can disturb the biosynthesis of progesterone of corpus luteum and reduce the serum progesterone level. Mitosis and decidulization of stroma cell can be disturbed owing to the low serum progesterone level, so the blastocyst cannot be implanted successfully.     

多氯联苯暴露对着床期子宫内膜组织特定基因甲基化的影响

目的:探讨多氯联苯(PCB118)慢性暴露对小鼠着床期子宫内膜特定基因启动子区甲基化水平及基因表达的影响。方法:将40只3周大CD-1小鼠随机分为4组:空白对照组,1μg/kg PCB118剂量组,10μg/kg PCB118剂量组,100μg/kg PCB118剂量组。各组灌胃1个月后,行促排卵合笼,于妊娠第4.5天断椎处死小鼠,留取子宫内膜组织。采用甲基化DNA免疫共沉淀-实时定量PCR(MeDIP-qPCR)检测特定基因启动子区甲基化水平;实时定量RT-PCR检测特定基因mRNA表达水平。结果:MeDIP-qPCR显示,相对于空白对照组,剂量组中PCDH17、ITGB8、FREM2及FGF4这4个基因启动子呈现高甲基化,与我们前期DNA甲基化芯片结果一致,有显著统计学差异(P<0.05)。Real-time PCR显示,PCDH17、ITGB8、FREM2及FGF4基因的mRNA水平在各剂量组均显著降低(P<0.05),与其启动子区DNA甲基化水平呈现负相关。结论:慢性PCB118暴露诱导着床期特定基因启动子区发生高甲基化,进而引起这些基因表达沉默,这可能是PCB118生殖毒性产生的分子机制之一。     

Effects of Acupuncture on Progesterone and Prolactin in Rats of Embryo Implantation Dysfunction

Objective:To investigate the effect of acupuncture on progesterone(P4) and prolactin(PRL) in rats of embryo implantation dysfunction(EID).Methods:On the first day of pregnancy,72 female Wistar rats were randomly allocated into the normal group,the EID model group,the acupuncture group and the P4 group(18 in each group).The normal group was injected sesame oil,while the other three groups were given mifepristone to establish the EID model.The acupuncture group and the P4 group were given treatment of acupuncture and P4 injection,respectively.The serum of P4 and PRL were detected by radioimmunoassay,and the mRNA and protein expressions of P4 receptor(PR) and PRL receptor(PRLR) were detected using real-time polymerase chain reaction and immunohistochemical method,respectively.Results:Compared with the normal group,the serum levels of P4 and PRL as well as the mRNA and protein expression levels of PR and PRLR in the EID model group were significantly lowered(P<0.01 or P<0.05).The above indices in the acupuncture group and the P4 group were significantly elevated compared with the EID model group(P<0.01 or P<0.05).Conclusion:Acupuncture can promote embryo implantation effectively,which might be related to the effects of acupuncture on upregulating the P4 and PRL levels in serum and the PR and PRLR expression levels in rats.     

Expression of TRAIL in Mouse Uterine Endometrium during Embryo Implantation

Tumor necrosis factor (TNF), which is primarily produced from activated macrophages, is one of the important cytokines that can induce cells to undergo apoptosis[1]. TNF-relatedapoptosis-inducing ligand (TRAIL), also called Apo-2L, is a new member of TNF …     

吲哚美辛对小鼠胚胎着床的影响

目的观察吲哚美辛对胚胎着床的影响,探讨吲哚美辛在胚胎着床中可能发挥的作用及其机制。方法小鼠妊娠第3天和第4天皮下注射吲哚美辛,观察其对胚胎着床的影响,经组织学切片,采用苏木精-伊红染色和透射电镜,观察用药后子宫内膜组织结构和细胞超微结构变化。结果①小鼠妊娠第3天和第4天皮下注射吲哚美辛50mg/kg可完全抗着床;②苏木精-伊红染色提示,给药后第5天和第8天小鼠子宫内膜基质致密,不见透明样改变;腺体增生,与基质发育不同步;血管明显减少;子宫内膜可见大量炎性细胞浸润;③超微结构示,给药后第5天小鼠子宫内膜全层脂滴,空泡变性呈不同程度增多,腔上皮细胞受损;第8天子宫内膜腺体少量细胞出现坏死。结论吲哚美辛通过多因素协同作用对小鼠具有较强抗着床作用。     

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.     

米非司酮对小鼠围着床期子宫内膜Dickkopf-1表达的影响

目的 研究孕酮抑制剂米非司酮对妊娠小鼠围着床期子宫内膜Dickkopf-1（DKK1）表达的影响，探讨DKK1在胚胎着床过程中的作用。方法 用半定量逆转录聚合酶链反应法（RT—PCR）及免疫组织化学SP法检测米非司酮组、载剂对照组和空白对照组小鼠在妊娠第3～7天DKK1 mRNA及蛋白的动态表达。结果载剂对照组和空白对照组小鼠在妊娠第4天出现DKK1 mRNA及蛋白的表达峰值，且两组在第3～7天各时间点DKK1的表达差异无显著性意义；米非司酮组DKK1 mRNA及蛋白的表达较两对照组显著降低（P〈0．01），在妊娠第4天亦无表达高峰。结论 DKK1是介导胚胎着床的重要因子之一，其表达可能受到孕激素的正向调节。     

不同针灸预处理对反复体外受精联合胚胎移植失败者子宫内膜容受性的影响

[目的]观察反复体外受精联合胚胎移植(IVF-ET)失败者接受3种不同针灸预处理后子宫内膜容受性(ER)的变化情况,寻找调整ER最佳的针灸预处理方法.[方法]将90例反复IVF-ET失败后低ER、未行胚胎移植(ET)患者随机分为针刺组、电针组和温针组,每组各30例.针刺组在月经干净后卵泡期开始行普通针刺治疗,至排卵期停...     

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)     

针灸对腹腔镜术后胃肠功能紊乱疗效的Meta分析

目的 评价针灸治疗对腹腔镜术后胃肠功能紊乱的疗效。方法 通过计算机检索中国期刊全文数据库（CNKI）、万方数据库（ Wanfang Data）、中国生物医学文献数据库（CBM）、重庆维普全文数据库（VIP）、PubMed、Embase等国内外主要数据库,纳入有关针灸治疗腹腔镜术后胃肠功能紊乱的随机对照试验（randomized clinical trials, RCT）,对纳入的文献进行资料提取和质量评价后采用RevMan 5.3统计软件进行Meta分析。结果 最终纳入13个RCT,共985例患者。Meta分析结果显示：与对照组比较（常规处理）,处理组（针灸治疗+常规处理）对腹腔镜术后胃肠功能恢复的有效率更高［RR=1.27,95%CI（1.14,1.42）,P＜0.000 1］;首次肛门排气时间更短［MD= －11.48,95%CI（－16.66,－6.31）,P＜0.000 1］;肠鸣音恢复时间更短［SMD=－2.36,95%CI（－3.02,－1.69）,P＜0.000 01］;首次肛门排便时间更短［SMD=－2.37,95%CI（－3.34,－1.40）,P＜0.000 01］。结论 针灸可以有效促进腹腔镜术后胃肠功能紊乱的恢复。但是,由于所纳入的文献证据质量等级不高,因此仍需要通过临床大样本、多中心、高质量的随机双盲试验来进一步补充验证。     

分期针刺规范化治疗中风后肢体功能障碍临床研究

目的：观察分期针刺配合康复训练治疗中风肢体功能障碍的临床疗效。方法：中风恢复期以肢体功能障碍为主的患者180名,随机分组为分期针刺＋康复组（A组）、传统针刺＋康复组（B组）、康复组（C组）。3组分别接受传统针刺的辨证取穴治疗和现代康复理论指导下的分期针刺取穴治疗,并同时接受系统现代康复治疗。分别应用美国国立卫生院卒中量表（NIHSS）、运动功能评定（FMA）、改良巴氏指数（MBI）以及偏瘫肢体Brunnstrom分期等量表进行疗效评定。结果：3组在总的临床疗效分期针刺＋康复组（A组）明显优于传统针刺＋康复组（B组）和康复组（C组）;在神经功能缺损量表、MBI量表、偏瘫肢体Brunnstrom分期量表积分治疗后经完全随机方差分析检验,分期针刺＋康复组（A组）明显优于传统针刺＋康复组和康复组（P〈0.001）,具有显著性差异。结论：分期针刺疗法配合康复治疗中风后偏瘫患者肢体功能运动及日常生活能力方面疗效显著。