Pituitary Suppression before Frozen Embryo Transfer Is Beneficial for Patients Suffering from Idiopathic Repeated Implantation Failure

Long-term gonadotropin-releasing hormone agonist（Gn RHa） administration before in vitro fertilization（IVF）/intracytoplasmic sperm injection（ICSI） in infertile women with endometriosis or adenomyosis significantly enhanced the chances of pregnancy in both fresh and frozen embryo transfer cycles. We hypothesized that long-term Gn RHa treatment might also be beneficial for the idiopathic repeated implantation failure（RIF） patients. In the 21 patients receiving Gn RHa and hormone replacement therapy（G-HRT） protocols for frozen embryo transfer, their data were compared with those of the 56 of frozen/fresh cycles they had previously undergone（previous protocols）. Comparison showed that the finial results were significantly better with G-HRT protocols than with their previous protocols, with pregnancy rate, clinical pregnancy rate, implantation rate and on-going pregnancy rate being 70%, 60%, 40% and 38% respectively with G-HRT protocols, against 17%, 11%, 6.3% and 5% with previous protocols. The results showed that hormonally controlled endometrial preparation with prior Gn RHa suppression could be used for patients who had experienced repeated failures of IVF treatment despite having morphologically optimal embryos, and the treatment may help increase the receptivity of the endometrium in these patients.

Clinical application of the endometrial microbiota test combined with personalized treatment in patients with repeated implantation failure

Objective:The objective of this study was to investigate the clinical value of the endometrial microbiota test(EMT)combined with personalized treatment in patients with repeated implantation failure(RIF).Methods:From March 2019 to August 2022,the clinical data of patients with RIF at the Reproductive Medicine Research Center of the Sixth Affiliated Hospital of Sun Yat-sen University were retrospectively analyzed.A total of 356 patients underwent the EMT using 16S rRNA sequencing and were included in the EMT group.The SPSS statistical software propensity score was set at 1:1 to match the age,anti-Müllerian hormone(AMH),antral follicle count(AFC),years of infertility,and body mass index(BMI)of the women who had not undergone the EMT;subsequently,354 patients were classified in the non-EMT group.We compared the clinical characteristics of the two groups and analyzed the clinical pregnancy outcomes after personalized treatment.Results:The clinical pregnancy rate(CPR)of the EMT group was significantly higher than that of the non-EMT group(47.81%vs.13.38%,P<0.05).Among 356 patients who underwent the EMT,55 patients(15.45%)in group A with normal endometrial microbiota composition did not test positive for the pathogenic bacteria causing chronic endometritis,34 patients(9.55%)in group B with abnormal endometrial microbiota composition did not test positive for the pathogenic bacteria causing chronic endometritis,35 patients(9.83%)in group C with normal endometrial microbiota composition tested positive for the pathogenic bacteria causing chronic endometritis,and 232 patients(65.17%)in group D with abnormal endometrial microbiota composition tested positive for the pathogenic bacteria causing chronic endometritis.After administration of probiotics or antibiotic plus probiotics treatment,the pregnancy rates of groups C and D were higher than those of group A(44.00%vs.20.00%and 55.03%vs.20.00%,respectively;bothP<0.05).Conclusion:The EMT combined with personalized treatment in patients with RIF has solid clinical value as a new treatment strategy.

Acupuncture improves the live birth of patients with repeated implantation failure: a retrospective cohort study

OBJECTIVE:To explore the therapeutic efficacy of acupuncture treatment on repeated implantation failure(RIF)patients with cryo-thawed embryo transfer(CET).METHODS:In a retrospective cohort study,all eligible women undergoing RIF were recruited in our center from January 1,2018 to December 31,2021.The patients were grouped by whether an acceptance of acupuncture treatment before CET,including the acupuncture group(Acu-group,55 cycles)and control group(Con-group,244 cycles).Data were analyzed by using binary logistic regression to explore the relationship of acupuncture treatment with pregnancy outcomes.RESULTS:The Acu-group had higher live-birth rate(LBR)[54.5%vs 41.0%,respectively;odds ratio(OR)=1.105,95%confidence interval(CI)(1.029,1.187),P=0.006]and ongoing pregnancy rate(OPR)[56.4%vs 43.0%,respectively;OR=1.100,95%CI(1.025,1.181),P=0.008]than the Con-group.There were no significant between-group differences in the rates of implantation[OR=1.070,95%CI(0.996,1.149),P=0.064],clinical pregnancy[OR=1.065,95%CI(0.997,1.138),P=0.061],biochemical pregnancy[OR=1.002,95%CI(0.903,1.112),P=0.967],or miscarriage[OR=0.778,95%CI(0.551,1.099),P=0.155].Perinatal outcomes did not differ significantly between the two groups.CONCLUSIONS:Acupuncture treatment could improve the LBR and OPR in RIF patients with CET cycles,suggesting a potential adjuvant therapy of acupuncture to improve the pregnancy outcomes in RIF patients.

Preimplantation genetic testing for aneuploidy improves clinical outcomes in patients with repeated implantation failure

Objective:The objective of this study is to study whether preimplantation genetic testing for aneuploidy(PGT-A)improves the clinical outcomes of infertile patients with repeated implantation failure(RIF)undergoing frozen-thawed embryo transfer.Methods:This is a retrospective analysis of clinical pregnancy,live birth,miscarriage rates,and obstetric and perinatal outcomes of women with RIF with or without PGT-A.Statistical analyses of categorical data were performed using propensity score matching(PSM),χ^(2)test,and Student’s t test.Results:We enrolled 466 patients with RIF,of which,209 were in the RIF-PGT-A group.The rate of euploid blastocysts was significantly associated with age and day 5 or 6 blastocysts.There were significant differences between the RIF-PGT-A group and the RIF-non-PGT-A group across several parameters.After PSM,positive serum human chorionic gonadotropin(56.9%and 33.9%,P<0.01),clinical pregnancy(49.5%and 31.2%,P<0.01),live birth(43.1%and 25.7%,P<0.01),and fetal heart rates(50.0%and 29.8%,P<0.01)per transfer were significantly higher in the RIF-PGT-A group.Conclusion:Elective single-embryo transfer PGT-A can minimize the risk of obstetric and perinatal outcomes,especially fetal body weight,in women with RIF.Additionally,PGT-A can significantly improve pregnancy and live birth rates.

The number and cytotoxicity and the expression of cytotoxicity-related molecules in peripheral natural killer(NK)cells do not predict the repeated implantation failure(RIF)for the in vitro fertilization patients

Natural killer(NK)cells are thought to play a key role in the successful establishment of a pregnancy by facilitating immunological adaptation of the semi-allogeneic developing embryo.The aim of this study was to explore the cell number,immunophenotypic characteristics,and activities of peripheral blood NK cells in women with repeated implantation failure(RIF).Peripheral blood was obtained from 27 women with RIF and 11 healthy,fertile controls during the middle luteal phase of the menstrual cycle.CD3^-CD56^+NK cells were quantified and analyzed by flow cytometry for the expression of cytolytic molecules(granzyme B,granulysin,and perforin)as well as cell surface receptors responsible for NK cell activation or inhibition(NKG2D,NKp30,NKp46,CD158a,CD158b).NK cytotoxicity was measured at three effector-to-target cell ratios.Women with RIF and fertile controls did not differ significantly in the percentage of circulating CD3CD56t NK cells,or in the proportions of these cells that expressed granzyme B,granulysin,or perforin.The two groups also did not differ significantly in the proportions of NK cells expressing the receptors NKG2D,NKp30,NKp46,CD158a or CD158b.General linear model analysis showed that NK cytotoxicity increased with effector-to-target cell ratio.However,NK cytotoxicity did not differ significantly between patients with RIF and fertile controls.These results suggest that RIF is not associated with significant alterations in the number or function of peripheral blood NK cells.

Endometrial MicroRNA Signature during the Window of Implantation Changed in Patients with Repeated Implantation Failure

Background： At present, a diagnostic tool with high specificity for impaired endometrial receptivity, which may lead to implantation failure, remains to be developed. We aimed to assess the different endometrial microRNA （miRNA） signatures for impaired endometrial receptivity by microarray analysis. Methods： A total of 12 repeated implantation failure （RIF） patients and I0 infertile patients, who conceived and delivered after one embryo transfer attempt, were recruited as RIF and control groups, respectively. Endometrial specimens from the window of implantation （WOI） were collected from these two groups. MiRNA microarray was conducted on seven and five samples from the RIF and control groups, respectively. Comparative, functional, and network analyses were performed for the microarray results. Quantitative real-time polymerase chain reaction （PCR） was performed on other samples to validate the expression of specific miRNAs. Results： Compared with those in the control group, the expression levels of 105 miRNAs in the RIF group were found to be significantly up- or down-regulated （at least 2-fold） by microarray analysis. The most relevant miRNA functional sets of these dysregulated miRNAs were miR-30 family, human embryonic stern cell regulation, epithelial-mesenchymal transition, and miRNA tumor suppressors by tool for annotations ofmicroRNA analysis. Network regulatory analysis found 176 miRNA-mRNA interactions, and the top 3 core miRNAs were has-miR-4668-5p, has-miR-429, and has-miR-5088. Expression levels of the 18 selected miRNAs in new samples by real-time PCR were found to be regulated with the same trend, as the result ofmicroarray analysis. Conclusions： There is a significant different expression of certain miRNAs in the WOI endometrium for RIF patients. These miRNAs may contribute to impaired endometrial receptivity.

Clinical Observations of Sequential Therapy with Chinese Medicine and Hysteroscopic Mechanical Stimulation of the Endometrium in Infertile Patients with Repeated Implantation Failure Undergoing Frozen-Thawed Embryo Transfer

Objectives： To investigate the clinical effect of sequential therapeutic intervention Yupei Qisun [compensating for weakness by invigorating Kidney （Shen） and Spleen （Pi） in advance] in Chinese medicine （CM） and hysteroscopic endometrial mechanical stimulation on the treatment of infertile patients with repeated implantation failure （RIF）; and to study the differences in patients＇ endometrial thickness and type on the day of embryo transfer, serum hormone levels on embryo transfer day and clinical pregnancy outcomes. Methods： In the clinical study, 168 frozen-thawed embryo transfer （FET） cycles for couples with RIF conforming to the research protocol were randomly divided into three groups： a CM group with 56 cycles （CM combined with FET）, a hysteroscopy group with 55 cycles （hysteroscopic endometfial mechanical stimulation）, and a control group with 57 cycles （conventional FET）. Differences in endometrial thickness on the embryo transfer day, levels of serum estradiol （E2） and progesterone （P） on the embryo transfer day, the E2/P ratio on the embryo transfer day, biochemical and clinical pregnancy rates, implantation rate, abnormal pregnancy rate and other indices were compared among the three groups. Results： Endometrial thickness, E2 and P levels, and the EJP ratio on embryo transfer day and other factors had no significant differences among groups. The biochemical pregnancy, clinical pregnancy, and implantation rates of the CM and hysteroscopy groups were significantly higher than the control group （P〈0.05）, and there were no significant differences between these two groups. The abnormal pregnancy rate had no significant difference among the three groups. Conclusions： Sequential therapy of Yupei Qisun could significantly improve the clinical outcomes of RIF-FET cycles, being equivalent to hysteroscopic endometrial mechanical stimulation, and provided a reliable method to treat such infertile couples.

抗核抗体阳性及干预治疗与反复种植失败辅助生殖结局相关性研究

目的探讨抗核抗体(ANA)阳性及干预治疗对反复种植失败(RIF)患者辅助生殖结局的影响。方法回顾性研究344例RIF患者,根据ANA检测结果分为ANA阳性组(294例)和阴性对照组(50例),抗核抗体阳性组分为ANA阳性低滴度组(214例)和ANA阳性高滴度组(80例)。比较阳性组和阴性组患者的一般临床资料、胚胎相关数据以及妊娠结局。采用Wilcoxon秩和检验、Mann-Whitney U检验、Kruskal-Wallis检验和卡方检验的统计学方法来分析ANA对RIF患者辅助生殖结局的影响,并分析了ANA阳性患者进行干预治疗后的临床妊娠情况。结果ANA阳性低滴度组和高滴度组患者临床妊娠率均显著低于阴性对照组(P<0.001);ANA阳性患者卵细胞受精率和卵裂率也均显著低于阴性对照组(P<0.05);因ANA阳性胚胎移植后未妊娠患者,经免疫调节剂治疗后的单周期临床妊娠率和累积临床妊娠率均明显提升(P<0.05)。结论与阴性对照组相比,ANA低滴度阳性和高滴度阳性患者的临床妊娠率均下降。经过临床干预治疗后,ANA阳性患者的单周期临床妊娠率和累积妊娠率得到了改善。这表明ANA阳性是RIF的重要原因,而免疫调节剂治疗是改善ANA阳性患者辅助生殖结局的有效措施。

反复种植失败患者子宫内膜CD138阳性对体外受精-胚胎移植妊娠结局的影响

目的:研究不明原因反复种植失败(RIF)患者子宫内膜CD138阳性对体外受精-胚胎移植周期临床妊娠结局的影响。方法:选取2019年7月至2023年5月温州医科大学附属第一医院收治的不明原因RIF患者332例(352个周期),行子宫内膜活检,免疫组化法查CD138阳性细胞。根据检测结果分成CD138阳性组和CD138阴性组,并按照每20个高倍镜视野(20 HPFs)下CD138阳性的数量分成1~5个/20 HPFs组,6~20个/20 HPFs组和>20个/20 HPFs组。对部分CD138阳性患者进行抗生素治疗,并随访妊娠结局。结果:在352个周期中,181个周期CD138检测阳性,阳性率为51.42%。其中1~5个/20 HPFs组有72个周期(39.78%),6~20个/20 HPFs组有63个周期(34.81%),>20个/20 HPFs组有46个周期(25.41%)。多因素Logistic回归分析结果显示,CD138阳性是影响临床妊娠的因素之一(OR=-0.474,95%CI=0.395~0.98,P=0.041)。CD138阳性组的临床妊娠率、胚胎种植率显著低于CD138阴性组(P<0.05)。当CD138阳性细胞数>20个/20 HPFs时,临床妊娠率和胚胎种植率显著低于CD138阴性组(P<0.05)。当C138个数少于5个/20 HPFs时,抗生素治疗和未治疗组妊娠结局差异无统计学意义(P>0.05)。CD138阳性患者抗生素治疗后持续阳性时,其临床妊娠率、胚胎种植率和活产率显著低于初检阴性和治疗后转阴患者(P<0.05);治疗后转阴患者和初检阴性患者的临床妊娠率、胚胎种植率和活产率差异无统计学意义(P>0.05)。结论:子宫内膜的CD138阳性细胞数>20个/20 HPFs降低了胚胎种植率、临床妊娠率,抗生素治疗后转阴可以改善妊娠结局,但持续阳性将影响妊娠结局。少量的CD138阳性(<5个/20 HPFs)对妊娠结局无明显影响。

子宫内膜种植窗检测结合个体化胚胎移植对反复种植失败患者的临床应用分析

目的:探讨子宫内膜种植窗检测(ERT)结合个体化胚胎移植(ET)改善反复种植失败患者的妊娠结局有效性。方法:回顾性分析2019年3月至2022年8月于中山大学附属第六医院生殖医学中心诊断为反复种植失败患者的临床资料,行ERT的428例作为ERT组,运用SPSS 26.0软件的倾向性评分匹配分析选择不行ERT的396例为对照组。比较两组的临床特征和妊娠结局,并比较子宫内膜种植窗(WOI)正常、延迟1 d、延迟2 d患者个体化ET后的妊娠结局。结果:ERT组临床妊娠率(47.73%vs.12.79%)和活产率(38.97%vs.9.76%)显著高于对照组,差异有统计学意义(P<0.05);并且ERT组的不孕因素中子宫内膜异位症(EMT)和多囊卵巢综合征(PCOS)因素占比较对照组高(P<0.05)。428例行ERT患者中发现有316例(73.83%)为WOI正常,有112例(26.17%)为WOI移位。WOI正常、WOI延迟1 d和WOI延迟2 d患者妊娠结局间差异无统计学意义(P>0.05)。结论:ERT技术结合个体化移植方案可以改善反复种植失败患者助孕结局,尤其是WOI移位患者,ERT技术具有应用前景和临床应用价值。

封闭抗体阴性的反复种植失败患者淋巴细胞主动免疫治疗助孕结局的影响因素分析

目的探究淋巴细胞主动免疫治疗(LIT)对反复种植失败的封闭抗体(BA)阴性患者助孕结局的影响及相关因素分析。方法选取2021年1月至2023年2月在我院自愿进行辅助生殖的BA阴性且反复种植失败患者84例作为研究对象。所有患者均接受LIT治疗,根据治疗后是否助孕成功划分为助孕成功组(n=58)和助孕失败组(n=26)。对比两组患者一般临床资料[年龄、配偶年龄、体质量指数(BMI)、流产次数、不孕持续时间、窦卵泡数]、LIT治疗相关特征(LIT制剂浓度、治疗次数、BA阳性率),以及治疗前性激素及抗苗勒管激素(AMH)水平、免疫细胞水平(CD4^(+)T细胞比例、CD8^(+)T细胞比例、CD4^(+)/CD8^(+)比值)及炎症因子水平[干扰素γ(IFN-γ)、白细胞介素4(IL-4)、白细胞介素10(IL-10)、白细胞介素17(IL-17)、转化生长因子β(TGF-β)]。通过Spearman相关性分析、多因素Logistic回归分析法筛选接受LIT治疗的反复种植失败BA阴性患者助孕成功的影响因素。结果两组患者一般临床资料及基础激素水平比较无统计学差异(P>0.05)。助孕成功组患者平均LIT制剂浓度及BA阳性率均显著高于助孕失败组患者(P<0.05);助孕成功组患者治疗前CD4^(+)T细胞比例及CD4^(+)/CD8^(+)比值均显著高于助孕失败组患者(P<0.05);助孕成功组患者治疗前血清IL-4、IL-10和TGF-β水平显著高于助孕失败组患者(P<0.05),而血清IL-17水平显著低于助孕失败组患者(P<0.05)。Spearman相关性分析及多因素Logistic回归分析表明,BA阳性、LIT制剂浓度较高、CD4^(+)T细胞比例较高、CD4^(+)/CD8^(+)比值较高、血清IL-4、IL-10、TGF-β水平较高、IL-17水平较低均是接受LIT治疗的反复种植失败BA阴性患者助孕成功的重要影响因素(P<0.05)。结论反复种植失败的BA阴性患者接受LIT可有效使BA转阳,其中使用LIT制剂浓度较高、促炎性细胞因子水平较低、抗炎性细胞因子水平及免疫细胞水平较高且BA转阳率较高的患者更容易在辅助生殖策略下助孕成功。

益气血补肝肾方对胚胎反复种植失败患者冻融胚胎移植周期妊娠结局的影响

目的探讨益气血补肝肾方对胚胎反复种植失败(RIF)患者冻融胚胎移植(FET)周期妊娠结局的影响。方法回顾性分析2017年1月至2023年6月在广州市第一人民医院生殖健康与不孕症专科行FET的120例RIF患者临床资料。患者月经周期规律,年龄25~35岁,按照FET前是否接受益气血补肝肾方治疗分为对照组和研究组,各60例;另选取同时期57例既往无胚胎移植史的FET患者作为正常组。3组均采用激素替代(HRT)方案准备内膜。研究组在准备内膜前先给予益气血补肝肾方3个月经周期(经后增殖方:煎成20 ml,制成颗粒剂10 g,1包/d,早晚分2次服用,月经干净后开始服用至排卵期;促黄体方:煎成20 ml,制成颗粒剂10 g,1包/d,早晚分2次服用,自排卵后开始服用至月经第1~3天停药)。比较3组患者内膜转化日子宫内膜厚度、血流搏动指数(PI)、动脉阻力指数(RI)、胚胎着床率和临床妊娠率。统计学方法采用单因素方差分析和χ^(2)检验。结果正常组和研究组子宫内膜厚度均高于对照组,PI、RI均低于对照组,3组子宫内膜厚度、PI、RI比较[(10.33±2.27)mm比(10.24±1.81)mm比(9.03±1.41)mm、(1.86±0.52)比(1.96±0.21)比(1.97±0.20)、(0.56±0.10)比(0.58±0.90)比(0.62±0.09)],差异均有统计学意义(F=8.40、6.08、6.89,均P<0.05)。正常组和研究组胚胎着床率、临床妊娠率均高于对照组,3组比较差异均有统计学意义[43.9%(43/98)比38.8%(40/103)比25.2%(26/103)、57.9%(33/57)比55.0%(33/60)比35.0%(21/60);χ^(2)=8.18、7.37,均P<0.05]。结论益气血补肝肾方可通过降低子宫内膜血流阻力指数和搏动指数,增加子宫内膜厚度,提高胚胎着床率和临床妊娠率,改善RIF患者FET周期妊娠结局。

宫腔灌注富血小板血浆对反复种植失败患者妊娠结局影响的Meta分析

目的评估宫腔灌注富血小板血浆(PRP)对反复种植失败(RIF)妇女妊娠结局的影响。方法使用PubMed、Embase、Scopus、Web of Science、CochraneLibrary、中国知识基础设施工程、维普数据库、万方数据库,检索PRP宫腔灌注治疗RIF患者的临床研究。提取数据后使用固定效应模型或随机效应模型进行Meta分析。结果共纳入7项研究。接受PRP宫腔灌注女性的生化妊娠率[OR=2.18,95%CI:[1.34,3.52],P<0.05]、临床妊娠率(OR=2.47,95%CI:[1.66,3.68],P<0.05)、活产率(OR=5.00,95%CI:[1.09,22.99],P<0.05)和胚胎种植率(OR=1.99,95%CI:[1.36,2.93],P<0.05)高于对照组,流产率低于对照组(OR=0.28,95%CI:[0.10,0.77],P<0.05)。亚组分析显示宫腔灌注0.5 mL PRP组的临床妊娠率高于宫腔灌注1.0 mL PRP组(P<0.05),流产率低于宫腔灌注1.0 mL PRP组(P<0.05)。结论宫腔灌注PRP可提高RIF患者的胚胎种植率、活产率、生化妊娠率和临床妊娠率,且显著降低流产的风险,宫腔灌注0.5 mL PRP具有更好的临床效果。

不孕症患者IVF-ET后不明原因反复种植失败的预测因素分析

目的分析不孕症患者体外受精-胚胎移植(IVF-ET)后不明原因反复种植失败(RIF)的预测因素。方法前瞻性纳入2020年6月至2022年6月无锡市妇幼保健院接诊的82例IVF-ET的不孕症患者作为研究对象,根据RIF发生原因将患者分为RIF组与不明原因RIF组。比较两组患者的基线资料、实验室指标,分析IVF-ET后不明原因RIF的影响因素;分析各项影响因素对不孕症患者IVF-ET后不明原因RIF的预测价值。结果82例患者中不明原因RIF 22例,占比为26.83%。Logistic回归分析结果显示,高水平CD4^(+)/CD8^(+)、纤维蛋白原(FIB)是不孕症患者IVF-ET后不明原因RIF的独立危险因素(OR>1,P<0.05),高水平抗米勒管激素(AMH)、血管内皮生长因子(VEGF)、粒细胞集落刺激因子(G-CSF)是其保护因素(OR<1,P<0.05)。绘制受试者工作特征(ROC)曲线结果显示,AMH、CD4^(+)/CD8^(+)、FIB、VEGF、G-CSF预测不孕症患者IVF-ET后不明原因RIF的曲线下面积(AUC)>0.70,均具有一定预测价值。结论AMH、CD4^(+)/CD8^(+)、FIB、VEGF、G-CSF对不孕症患者IVF-ET后不明原因RIF有一定预测价值。

配偶同步认知疗法对反复胚胎种植失败病人受孕结局及其心理状况的影响

目的:探讨配偶同步认知疗法对反复胚胎种植失败(RIF)病人的影响。方法:选取医院2020年7月—2022年7月收治的136例RIF病人为研究对象,按入院建档顺序分为对照组和观察组,每组68例。对照组采用常规护理,观察组采用配偶同步认知疗法,比较两组受孕结局、干预前后心理状况[正性情绪(PA)、负性情绪(NA)]、自我效能感[一般自我效能感评分(GSES)]、生育生活质量[生育生活质量评分(FertiQoL)]。结果:观察组胚胎种植率、临床妊娠率高于对照组(P<0.05),自然流产率低于对照组(P<0.05);干预后观察组PA评分、GSES评分高于对照组(P<0.05),NA评分低于对照组(P<0.05);干预后观察组FertiQoL评分高于对照组(P<0.05)。结论:配偶同步认知疗法能提高RIF病人的受孕率,改善其不良心理状态,提升病人的自我效能感,提高病人的生活质量。

不同子宫内膜种植窗检测方法对反复种植失败患者妊娠结局的影响

目的:采用子宫内膜容受性阵列(ERA)测序或子宫内膜组织学检测方法评估反复种植失败(RIF)患者子宫内膜种植窗,探讨两种技术对于改善此类患者临床结局的有效性及成本效益分析。方法:回顾性队列研究2018年1月至2022年12月在甘肃省妇幼保健院生殖医学中心诊断为RIF的125例患者的临床资料。根据是否接受子宫内膜容受性检测和使用检测技术不同,分为对照组(n=36)、基因组(n=35)和组织学组(n=54),比较3组临床资料和妊娠结局。结果:①单因素方差分析结果显示:基因组和组织学组患者的胚胎植入率显著高于对照组,差异有统计学意义(P<0.05),基因组与组织学组患者的胚胎植入率差异无统计学意义(P=0.48);②3组患者的临床妊娠率及活产率差异均无统计学意义(P>0.05);③Log-rank检验显示:基因组和组织学组患者50%达活产时间显著小于对照组,差异有统计学意义(P<0.05),基因组与组织学组50%患者达活产时间差异无统计学意义(P>0.05);④对照组患者平均移植胚胎数显著高于基因组和组织学组,差异有统计学意义(P<0.05),基因组患者花费成本显著高于组织学组,差异有统计学意义(P<0.05)。结论:①对于RIF的患者可行子宫内膜种植窗检测,可有效缩短达活产时间,减少移植胚胎数目;②ERA测序和子宫内膜组织学检测作为评估子宫内膜种植窗的方法均具有局限性,尚不能明确哪种检测方法在准确性和实用性方面更具有优势。

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、接受优质囊胚移植、女方年龄4个变量是获得活产的独立影响因素(P<0.05)。结论联用ERT与PGT是治疗RIF的一次成功尝试,可在单独应用ERT、PGT的基础上进一步改善RIF患者的FET结局。

反复种植失败患者阴道菌群特征研究

目的:探讨反复种植失败(repeated implantation failure,RIF)患者的阴道菌群特征。方法:纳入49例接受冻融胚胎移植(frozen-thawed embryo transfer,FET)患者。其中观察组患者(n=29)有RIF病史且本次FET周期妊娠失败;对照组患者(n=20)首次行FET且成功妊娠。2组均采取人工周期准备内膜、移植第5天优质囊胚。于FET日收集患者阴道分泌物,通过16S rRNA测序对2组阴道菌群多样性、群落组成以及差异菌属进行分析。用受试者工作特征(receiver operating characteristic,ROC)曲线评估阴道乳杆菌属丰度诊断RIF的效能。结果:α多样性分析显示,观察组阴道菌群多样性(Shannon指数:P=0.0498)和丰富度(Chao指数:P=0.0471)均显著高于对照组。β多样性分析显示,2组阴道菌群结构差异有统计学意义(P=0.016)。在属水平上,2组阴道的优势菌均为乳杆菌属,与对照组相比,观察组乳杆菌属丰度显著降低,而不动杆菌属和假单胞菌属丰度显著增加(LDA>3,P<0.05)。阴道乳杆菌属丰度诊断RIF的最佳截断值为70%,曲线下面积(area under curve,AUC)为0.77。结论:RIF患者阴道菌群显示差异,以乳杆菌属丰度降低为主要特征。阴道乳杆菌属低丰度可能与RIF相关。

反复种植失败患者卵裂胚解冻培养囊胚移植的效果分析

目的将反复种植失败(repeated implantation failure,RIF)患者中剩余卵裂胚解冻并培养囊胚移植作为一种潜在的治疗策略,并分析其妊娠结局。方法本研究纳入295名RIF患者进行冻融胚胎移植,根据囊胚来源分为卵裂胚解冻培养囊胚移植组(n=141)和传统解冻囊胚移植组(n=154)。研究两组患者临床妊娠率、种植率和活产率的区别。结果卵裂胚解冻培养囊胚移植组的临床妊娠率、种植率均明显高于传统解冻囊胚移植组(P<0.05),两组之间的流产率、多胎率和活产率差异无统计学意义(P>0.05)。RIF≤4次和RIF=5次的患者,卵裂胚解冻培养囊胚移植组的种植率高于传统的解冻囊胚移植组(P<0.05),而RIF≤4次,RIF=5次和RIF≥6次的患者,两组临床妊娠率、流产率、多胎率和活产率差异均无统计学意义(P均>0.05)。结论当RIF患者囊胚为零时,解冻剩余的卵裂胚并培养囊胚进行移植可以提高临床妊娠率和种植率,是一种有效治疗策略。

不孕症子宫内膜类器官建立与验证

目的 构建非免疫因素致不孕症和免疫因素致不孕症子宫内膜类器官培养体系,并比较两者免疫细胞因子的差异。方法 标本采自海军军医大学(第二军医大学)第一附属医院生殖医学中心行宫腔镜检查的不孕症患者,获取非免疫因素致不孕症患者(n=3)与反复着床失败(RIF)患者(n=5)子宫内膜组织,通过洗涤、消化、重悬及接板,将组织嵌入基质胶中进行3D培养;在倒置显微镜下观察两组子宫内膜类器官生长情况;利用免疫荧光染色技术检测子宫内膜特异性标志物雌激素受体(ER)、角蛋白和上皮钙黏蛋白表达情况;利用ELISA法测定两组细胞因子水平,并比较两组细胞因子的差异。结果 在子宫内膜类器官体外培养过程中,类器官体积逐渐增大、细胞数量逐渐增多,培养7~10 d的类器官体积达到稳定状态且形状逐渐趋于圆形。在相同时间内,非免疫因素致不孕症子宫内膜类器官数量较RIF子宫内膜类器官多。免疫荧光染色显示类器官中子宫内膜相关标志蛋白ER、角蛋白和上皮钙黏蛋白均表达,提示子宫内膜类器官构建成功。ELISA检测结果显示,两组间干扰素γ、IL-10、TNF-α、IL-4水平和TNF-α/IL-4比值的差异均有统计学意义(P<0.05或P<0.01),TGF-β1、IL-17水平及IL-17/TGF-β1、干扰素γ/IL-10、TNF-α/IL-10比值差异均无统计学意义(均P>0.05)。结论 在体外成功培养获得了具有增殖能力的非免疫因素致不孕症与RIF子宫内膜类器官,为免疫性不孕症的基础研究提供了新模型。