反复胚胎种植失败母体因素分析及治疗

反复胚胎种植失败(RIF)是人工辅助生殖技术中的难题,影响RIF的原因主要分为胚胎因素及母体因素,其中母体因素主要包括宫腔内占位性病变、输卵管积水、子宫腺肌症、子宫内膜炎症等母体病理因素,以及免疫因素、血液高凝状态、子宫收缩过频及子宫内膜容受性不良等。本文从引起RIF的母体因素角度进行综述,针对不同母体因素总结目前的治疗方法进展。     

反复种植失败的治疗新进展

反复种植失败(RIF)是指患者连续经过多次胚胎移植,仍无法正常妊娠.RIF可以归因于母体因素和胚胎因素,母体因素也可分为局部子宫内膜容受性和全身因素.根据病因,RIF的临床治疗主要包括子宫内膜容受性的改善、全身因素的纠正包括高凝状态的改善和免疫治疗,以及提高胚胎种植率等.目前RIF的治疗较多是围绕子宫内膜容受性的提高,...     

Th17及Treg相关因子在反复种植失败患者种植窗口期子宫内膜中的表达

目的通过研究种植窗口期子宫内膜中Treg及Th17特异性转录因子及相关细胞因子表达情况,探讨反复种植失败患者子宫内膜中Treg/Th17平衡调节的可能机制。方法建立种植窗口期子宫内膜标本库。从标本库选择2013年10月至2014年12月收集的符合实验要求的反复种植失败患者(实验组)及首次移植妊娠患者(对照组)各20例,采用定量RT-PCR测量内膜标本中的Treg、Th17特异性表达因子Foxp3和ROR-γt及相关细胞因子IL-10、IL-17表达情况。结果在实验组和对照组子宫内膜上均可见Foxp3、ROR-γt及细胞因子IL-10、IL-17表达;与对照组比较,实验组Foxp3mRNA的相对表达量[(0.77±0.15)vs.(1.57±1.22)]及IL-10mRNA的相对表达量[(1.14±1.03)vs.(1.54±1.77)]均显著降低(P0.05),而ROR-γt mRNA的相对表达量[(0.92±0.39)vs.(0.55±0.15)]及IL-17mRNA的相对表达量[(0.95±0.31)vs.(0.49±0.21)]均显著升高(P0.05)。结论 IL-17、IL-10可能参与内膜中Th17/Treg平衡调节。     

宫腔灌注粒细胞集落刺激因子在不明原因反复种植失败患者中的应用评价

目的 研究宫腔灌注粒细胞集落刺激因子(G-CSF)对不明原因反复种植失败(URIF)患者的子宫内膜容受性及妊娠结局的影响.方法 采用前瞻性随机对照的研究方法,选择2019年1月至2020年9月期间于大连市妇女儿童医疗中心就诊的,拟行人工周期-冻融胚胎移植的U RIF患者120例,将患者随机分为试验组和对照组(每组各60...     

FKBP52在反复体外受精种植失败患者种植窗期子宫内膜的表达研究

目的探讨FKBP52在反复体外受精种植失败患者种植窗期子宫内膜的表达情况。方法采用实时荧光定量聚合酶链式反应(RT-PCR)技术和免疫组织化学方法分别测定20例反复体外受精种植失败妇女(组1)种植窗期子宫内膜组织中FKBP52蛋白及其mRNA的表达,并与15例正常生育能力妇女卵泡晚期(组2)和种植窗期(组3)相比较。结果组1的FKBP52mRNA表达水平低于组2,组2低于组3,但差异均无统计学意义(P0.05)。FKBP52蛋白表达于腺上皮的胞核及胞浆,组1在腺上皮的胞核表达明显,组2在腺上皮的胞浆表达明显,组3在腺上皮的胞核及胞浆均有明显表达;组1和组2的FKBP52蛋白表达量分别为(2 125.9±1 729.9)和(1 360.2±853.7),并显著低于组3(8 319.1±5 873.8)(P0.05,P0.01)。结论 FKBP52蛋白在种植窗期的表达和定位异常可能是反复体外受精种植失败患者子宫内膜容受性的影响因素。     

反复种植失败妇女“种植窗”时期子宫内膜组织的细胞骨架蛋白质的差异表达

目的探讨体外受精-胚胎移植治疗周期中反复种植失败(RIF)和已生育正常妇女"种植窗"时期(LH+6~LH+9d)子宫内膜组织蛋白质表达的差异。方法选择RIF妇女和正常对照妇女各4名分为实验组和对照组。采用胶内差异双向电泳、(2D-DIGE)和基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)技术鉴定子宫内膜组织的差异表达蛋白质。结果两组之间获得16个差异表达蛋白质(其中6个下调表达,10个上调表达),其中10个参与细胞骨架构成,分别是:波形蛋白、埃兹蛋白、Septin 2蛋白、radixin蛋白、纽蛋白、角蛋白8、角蛋白10、角蛋白18、角蛋白19及肌动蛋白。结论本研究结果证实"种植窗"时期RIF妇女子宫内膜组织存在多个细胞骨架蛋白质表达的异常,可能是子宫内膜细胞"质膜转换"缺陷和胚胎种植失败的重要原因。     

子宫内膜容受性检测联用胚胎植入前遗传学检测在反复种植失败患者中的效果研究

目的 评估联用子宫内膜容受性检测(ERT)与胚胎植入前遗传学检测(PGT)技术在反复种植失败(RIF)患者冻融胚胎移植(FET)周期中的应用效果。方法 选取2018年1月至2023年1月在徐州市妇幼保健院生殖医学中心接受FET的RIF患者,根据患者接受的评估检测不同分为4组：接受ERT评估(基于转录组测序的ERT技术模型)后移植经PGT检测筛选的整倍体囊胚RIF患者(联用组,n=138)、均未接受ERT评估和PGT检测移植RIF患者(RIF组,n=324)、仅接受ERT评估后移植RIF患者(ERT组,n=147)和仅接受PGT检测筛选整倍体囊胚移植RIF患者(PGT组,n=121),比较4组患者的基本情况及FET结局,并对RIF患者活产率的影响因素进行Logistic回归分析。结果 95.79%(273/285)接受ERT评估的RIF患者的内膜容受期会推迟1～2 d,根据ERT评估结果相应调整了移植时间。各组患者的FET结果显示,联用组的胚胎种植率、宫内妊娠率、活产率均显著高于PGT组、ERT组与RIF组(P<0.05)。Logistic回归分析结果表明：实施PGT、接受ERT、...     

子宫内膜容受性检测在反复种植失败患者体外受精-胚胎移植中的应用

目的 评估子宫内膜容受性检测(ERT)技术在反复种植失败(RIF)患者冻融胚胎移植(FET)周期中的应用效果。方法 将接受ERT评估(基于转录组测序的ERT技术模型)的RIF患者(ERT组,n=60)与未接受ERT评估的RIF患者(非ERT组,n=151)、非RIF患者(非RIF组,n=503)的FET结局进行比较。结果 接受ERT评估的RIF患者中绝大部分(58/60,96.67%)的内膜容受期推迟了1～2 d,根据ERT评估结果相应调整了移植时间。FET结果显示,与非ERT组比较,ERT组胚胎种植率(53.25%vs. 35.07%)、宫内妊娠率(66.67%vs. 45.70%)、活产率(55.00%vs. 29.80%)均显著升高(P<0.05),且与非RIF组相当(P>0.05)。不同移植策略(1～2枚卵裂期胚胎、1枚囊胚移植、1枚卵裂期胚+1枚囊胚序贯移植)下,ERT组宫内妊娠率、活产率均显著高于非ERT组(P<0.05)。结论 对RIF患者进行ERT,调整移植时间进行移植,可显著改善RIF患者的FET结局。     

反复种植失败患者微刺激方案IVF-ET助孕成功一例及文献复习

反复种植失败是体外受精-胚胎移植失败的主要原因之一,尽管其定义及诊疗尚存在争议,但对于反复种植失败的患者应当尽可能找到原因,针对原因进行相应的处理已成为共识。导致反复种植失败的原因可概括为包括卵母细胞质量、精子质量、胚胎遗传物质异常等在内的胚胎质量因素和包括母体解剖结构异常、内膜局部或全身免疫失调、代谢紊乱等在内的母体因素,此外,还存在部分原因不明的反复种植失败,其临床处理办法仍在探索和尝试。本文报道微刺激方案帮助1例2次控制性促排卵IVF、8次移植的反复种植失败患者成功生育的过程,并结合其病史、IVF-ET助孕经过及妊娠结局,对反复种植失败的定义、病因及临床治疗等相关文献进行整理和总结,探讨该患者成功妊娠的可能原因,为反复种植失败患者的诊疗提供新思路。     

反复种植失败患者冻融胚胎移植的三种内膜准备方案的分析

目的使用GnRH-a 3.75mg降调节激素替代、单纯激素替代及自然周期内膜准备方案对提高反复种植失败患者冻融胚胎移植(FET)妊娠率的利弊分析。方法选取160例(周期)行FET的反复种植失败患者,根据内膜准备方式分为自然周期组(A组)82例、激素替代周期组(B组)36例和降调节激素替代组(C组)42例。比较三组患者的种植率、妊娠率、流产率和生殖激素水平。结果A、B和C三组患者的种植率分别为23.61%、21.25%和38.78%,临床妊娠率分别为42.68%、36.10%和61.90%;C组与A组、B组的种植率和临床妊娠率比较差异均有统计学意义(P<0.05)。A组与B组间胚胎移植日内膜厚度差异有统计学意义[(10.40±1.81)mm vs.(9.12±1.61)mm](P<0.05)。三组患者的流产率和异位妊娠率比较,差异无统计学意义(P>0.05)。结论与自然周期内膜准备方案及传统激素替代内膜准备方案比较,3.75mg GnRH-a降调节激素替代内膜准备方式可提高反复种植失败患者冻融胚胎移植妊娠率而不增加流产率。     

易栓症与反复胚胎种植失败的相关性研究

目的通过对反复胚胎种植失败(RIF)患者易栓症标志物进行筛查,评估其发病规律及高危因子,提供有效的预防及治疗方案。方法应用测序技术、血液凝固法、发色底物法及酶联免疫吸附法检测90例RIF患者(病例组)及90例首次胚胎移植即成功妊娠的不孕症患者(对照组)血浆中亚甲基四氢叶酸还原酶多态性(MTHFR C677T)、蛋白S活性、蛋白C活性、抗凝血酶(AT-Ⅲ)活性、抗心磷脂抗体(ACL-IgG/IgM)、抗β2糖蛋白I抗体(抗β2-GPI)的变化。结果 MTHFR C677T位点T/T基因型在病例组显著高于对照组(43.3%vs.23.3%,P0.05),Cramer V相关系数为0.574(P0.05);病例组与对照组ACL-IgG/IgM阳性率、抗β2-GPI阳性率、蛋白S活性、蛋白C活性、抗凝血酶活性均无显著差异(P0.05)。结论 MTHFR C677T位点T/T基因型可能与RIF的发生有关,可作为本地区IVF患者的易栓症筛查指标。     

Downregulation of T helper type 1 immune response and altered pro-inflammatory and anti-inflammatory T cell cytokine balance following conventional but not laparoscopic surgery

BACKGROUND: The clinical advantages of laparoscopic procedures result from a minimized surgical trauma. The present study was performed to investigate immunosupression following laparoscopic operations as compared with open surgery. Our analysis focused on the T cell secretion of cytokines that regulate the critical balance of either T helper type-1 (Th1)- and Th2-mediated immune responses on pro- and antiinflammatory activities. METHODS: In a prospective study, immunological data of 26 patients submitted to laparoscopic cholecystectomy (LCE) and 17 patients undergoing conventional cholecystectomy (CCE) for symptomatic cholecystolithiasis were compared. Patients with acute cholecystitis and patients developing postoperative complications or receiving immunosuppressive medication were excluded. Production of interferon (IFN)-gamma, interleukin (IL)-2, IL-4, tumor necrosis factor (TNF)-alpha, and IL-10 by isolated T cells stimulated by cross-linking of CD3 and CD28 was evaluated preoperatively as well as on postoperative days 1 and 6 or 7. Cytokines were measured by immunoenzymometric assay. RESULTS: IFN-gamma, TNF-alpha, and IL-2 production by T cells decreased significantly by 48.3%, 36.6%, and 36.8%, respectively, on postoperative day 1 after CCE, but not after LCE. These results indicate severe suppression of Th1-type and proinflammatory cytokines after the open operation. In contrast, IL-4 and IL-10 did not show significant changes in either group suggesting that Th2 cell response and anti-inflammatory activity remained normal. CONCLUSIONS: The present study shows that open, but not laparoscopic cholecystectomy is associated with a marked suppression of T lymphocytes functions as indicated by deregulation of both the Th1/Th2 and the pro-/anti-inflammatory cytokine balance. The results therefore suggest that downregulation of Th1 cell-mediated immune response and pro-inflammatory activity of T cells is a hallmark of open, but not laparoscopic surgery.     

Sperm DNA fragmentation,recurrent implantation failure and recurrent miscarriage

Evidence is increasing that the integrity of sperm DNA may also be related to implantation failure and recurrent miscarriage (RM). To investigate this, the sperm DNA fragmentation in partners of 35 women with recurrent implantation failure (RIF) following in vitro fertilization, 16 women diagnosed with RM and seven recent fathers (control) were examined. Sperm were examined pre- and post-density centrifugation by the sperm chromatin dispersion (SCD) test and the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. There were no significant differences in the age of either partner or sperm concentration, motility or morphology between three groups. Moreover, there were no obvious differences in sperm DNA fragmentation measured by either test. However, whilst on average sperm DNA fragmentation in all groups was statistically lower in prepared sperm when measured by the SCD test, this was not seen with the results from the TUNEL assay. These results do not support the hypothesis that sperm DNA fragmentation is an important cause of RIF or RM, or that sperm DNA integrity testing has value in such patients. It also highlights significant differences between test methodologies and sperm preparation methods in interpreting the data from sperm DNA fragmentation tests.     

反复植入失败原因及处理对策

体外受精-胚胎移植（IVF-ET）的成功率受到胚胎质量、宫腔内环境、免疫等多种因素的影响,任何环节出现异常都有可能导致整个治疗周期的失败.反复植入失败（RIF）不但加重了患者的经济负担,而且对其身心亦造成了双重打击.为此,积极探寻RIF原因,采取有效的治疗措施降低IVF失败率,从而改善IVF结局成为辅助生殖技术中亟待解决的问题.     

淋巴细胞免疫治疗在反复种植失败和复发性自然流产中的作用

<正>现代免疫学观点认为,正常妊娠时母一胎界面表现为一种特殊类型的外周免疫耐受机制,这种耐受状态形成复杂,各种免疫因素通过有机协调形成网络,达到母胎间免疫平衡,使妊娠得以维持。成功的妊娠本身就是一个免疫矛盾所在。一方面胎儿抗原不能作为"半同种移植物"被母体识别,另一方面为了在母体中生存,胎儿抗原又必须呈递给母体,被母体接受。因此,胎儿抗原的免疫识别对于维持妊娠     

Regulation of pro-inflammatory and anti-inflammatory cytokine responses by Kupffer cells in endotoxin-enhanced reperfusion injury after total hepatic ischemia

The effects of Kupffer cells on cytokine responses in endotoxin-enhanced reperfusion injury after total hepatic ischemia were investigated in this study. Male rats pretreated with either normal saline solution (NS group) or gadolinium chloride (GdCl(3)) to inhibit Kupffer cell function (GC group) were subjected to 60 min of hepatic ischemia. These animals received either normal saline solution or sublethal doses of endotoxin (1 mg/kg) at reperfusion. In the NS group, endotoxin administration induced an enhanced tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 production 1 h after reperfusion with a subsequent peak of macrophage inflammatory protein-2 (MIP-2) levels, which resulted in a 7-day survival rate of 30%. Despite endotoxin administration, GdCl(3) pretreatment significantly suppressed TNF-alpha and increased interleukin-10 production 1 h after reperfusion, which led to a decline in MIP-2 production and amelioration of functional and structural liver damage with a 7-day survival rate of 80%. Augmented pro-inflammatory and anti-inflammatory cytokine responses by Kupffer cells were associated with endotoxin-enhanced reperfusion injury after hepatic ischemia. Kupffer cell blockade has a potential to attenuate the insult via modulation of cytokine responses.     

反复种植失败患者的个体化胚胎移植

反复种植失败(recurrent implantation failure,RIF)是目前辅助生殖技术(assisted reproductive technology,ART)临床工作中的难题,也是研究和讨论的热点,同时也给患者带来了巨大的经济负担及精神压力。其定义目前尚未统一。结合胚胎移植次数、移植胚胎数目和质量、子宫内膜容受性等来定义RIF可能更准确和全面。由于种植窗不是恒定不变的,统一的胚胎移植时间可能导致子宫内膜与胚胎发育不同步,从而出现RIF。因此个体化胚胎移植是改善RIF患者ART结局的重要手段。     

Elevated sperm DNA fragmentation does not predict recurrent implantation failure

In this study, we sought to determine whether sperm DNA fragmentation (DFI%) and high DNA stainability (HDS%) evaluated by sperm chromatin structure assay (SCSA) predict recurrent implantation failure (RIF) or pregnancy rate. A retrospective study was performed of consecutive cycles of ICSI treatment from 2009 to 2018. A total of 386 couples that underwent 1,216 frozen embryo transfer (FET) cycles were analysed. Mean female and male age was 34 ± 3.6 years and 37.3 ± 6.6 years, respectively, and a median total motile sperm count (TMSC) was 43.5 [9.9–105.5] million. Overall median DFI% and HDS% was 12 [7.1–18.9] and 9.6 [6.5–14.4] respectively. On multivariable analysis, DFI% and HDS% were not associated with RIF (DFI%: OR = 1.01, 95% CI: 0.98–1.04, p = .414; HDS%: OR = 0.97, 95% CI: 0.94–1.01, p = .107) or IVF success, defined as clinical pregnancy (DFI%: OR = 1.00, 95% CI: 0.99–1.01, p = .641; HDS%: OR = 1.01, 95% CI: 0.99–1.02, p = .565). We found that neither DFI% or HDS%, as assessed by SCSA, were predictive of RIF or pregnancy rate. This finding suggests that sperm DNA fragmentation does not predict RIF or pregnancy rate.