精子DNA损伤对IVF/ICSI-ET结局影响的回顾性资料分析

目的:探讨精子DNA损伤对体外受精/卵胞浆内单精子注射-胚胎移植(IVF/ICSI-ET)结局的影响。方法:回顾性分析了初次行常规IVF和ICSI治疗的192例不孕夫妇的临床资料。分别比较行IVF和ICSI治疗时,精子DNA碎片指数(DFI)≤15%和DFI15%的受精率、优质胚胎率、种植率、临床妊娠率和流产率的差异;并比较DFI15%时,不同受精方式(IVF或ICSI)对受精率、优质胚胎率和临床妊娠率的影响。结果:在IVF或ICSI治疗周期中,精子DNA损伤对受精率、优质胚胎率、种植率、临床妊娠率和流产率均无影响(P0.05)。在DFI15%情况下,行IVF治疗的受精率、优质胚胎率、种植率和临床妊娠率与行ICSI治疗比较,无统计学差异(P0.05)。结论:精子DNA损伤不影响IVF/ICSI-ET的结局。对于DFI15%的患者ICSI-ET与IVF-ET的治疗结局相似。     

同源卵子行分半IVF/ICSI比较

目的:比较同源卵子行分半IVF/ICSI时的受精率、卵裂率、可用胚胎数及优胚数。方法:回顾性分析2007年8月～2011年8月在石家庄市第四医院生殖医学中心接受辅助治疗第一周期的患者,行分半IVF/ICSI 106周期,根据IVF受精情况分为4组:①组IVF均未受精;②组IVF受精率低下(受精率<25%);③组IVF受精率为25%～50%;④组IVF受精率>50%。比较4组同源卵子IVF及ICSI受精率、卵裂率、可用胚胎数及优胚数。结果:共行分半IVF/ICSI 106周期,常规IVF 392周期,均为第一周期。分半IVF/ICSI和常规IVF 4组率的比较,差异有统计学意义。①组和②组因IVF受精率低,ICSI可用胚胎数及优胚数均高于IVF;④组IVF可用胚胎数、优胚数均高于ICSI组;③组IVF、ICSI可用胚胎数及优胚数比较差异无统计学意义。结论:分半IVF/ICSI可避免IVF受精失败或受精低下导致无可用胚胎,但同源卵子IVF、ICSI均受精时,ICSI对精子、卵子的操作导致其优胚率低于IVF受精的优胚率。     

精子DNA损伤对IVF-ET中胚胎质量及临床结局影响

目的:探讨精子DNA损伤对体外受精-胚胎移植(IVF-ET)中胚胎及临床结局影响。方法:选择2015年1月—2018年7月本院行IVF治疗、女方≤35岁的夫妇2008对进行临床分析。所有男性患者均行精液常规检查以及精子DNA检查。按照DNA受损精子所占百分比(DFI)分为DFI 15%组90例、15%～30%组1831例、≥30%组87例,比较3组基本情况、精液常规、胚胎培养和临床结局。结果:DFI 15%组、DFI 15%～30%组男性精子密度、精子活率、精子正常形态率均高于DFI≥30%组(P0.05);3组女方年龄、男方年龄、不孕时间、女方基础卵泡刺激素、基础黄体生成素、用药Gn总量和总天数无差异,平均获卵数、成熟卵率、受精率、正常受精率、卵裂率、优胚率和囊胚形成率无差异(均P0.05)。DFI15%组90例中55例、DFI 15%～30%组1831例中1154例和DFI≥30%组87例中54例进行了新鲜胚胎移植,3组平均移植胚胎数、移植卵裂期胚胎率、移植囊胚率、临床妊娠率、种植率和流产率等无差异(P0.05)。Pearson分析显示,DFI与体质指数(r=0.314,P=0.722)及年龄(r=0.159,P=0.873)均无相关性。结论:精子DNA损伤会影响精子质量,但对IVF-ET培养的胚胎质量及临床结局无影响。     

隐匿精子症和无精子症卵胞浆内单精子注射的临床结局比较

目的:比较隐匿精子症和无精子症卵胞浆内单精子注射(ICSI)的临床结局.方法:选择1999 ～ 2012年在唐山市妇幼保健院生殖遗传科的156例隐匿性精子症和288例无精子症患者,无精子症患者分别通过附睾或睾丸穿刺取精后进行ICSI治疗,比较受精率、卵裂率、优胚率、生化妊娠率、临床妊娠率、流产率、出生缺陷率等.结果:三组的受精率、卵裂率差异有统计学意义(P＜0.05),优胚率、生化妊娠率、临床妊娠率、流产率、出生缺陷率差异无统计学意义(P＞0.05).结论:隐匿精子症和无精子症ICSI后虽然受精率、卵裂率不同,但临床结局相同.     

精子DNA损伤对常规体外受精-胚胎移植结局的影响

目的探讨精子DNA损伤对精液参数及常规体外受精-胚胎移植(in vitro fertilization and embryo transfer,IVF-ET)结局的影响。方法选择2012年10月至2014年11月在四川省妇幼保健院行IVF-ET的169对不孕夫妇的资料,用精子染色质扩散实验(sperm chromatin dispersion,SCD)测定男性患者精子DNA碎片率(DNA fragmentation index,DFI),同时行常规精液分析;按DFI值将男性患者分为A组(DFI30%)148例和B组(DFI≥30%)21例,比较两组精液参数、受精率、正常受精率、卵裂率、可利用胚胎率、优质胚胎率、临床妊娠率和着床率。结果 B组男性平均年龄高于A组(P0.05);B组精子浓度、前向运动精子百分比、正常形态率、受精率、正常受精率均显著低于A组(P0.05);卵裂率、可利用胚胎率、优质胚胎率、临床妊娠率、着床率两组比较差异无统计学意义(P0.05)。结论精子DNA损伤随男性年龄增加而增加,与精子浓度、前向运动精子百分比、正常形态率有关,对IVF-ET中的受精情况有负面影响。     

精子DNA碎片指数对体外受精-胚胎移植结局的影响

目的:探究精子DNA碎片指数(DFI)对体外受精-胚胎移植(IVF-ET)结局的影响。方法:选取2015年9月-2018年9月本院生殖医学中心IVF-ET助孕初筛患者486对,精子染色质结构分析法联合流式细胞术检测患者精子DAN碎片,根据DFI分为DFI≤15%组、15%DFI≤30%组、DFI30%组,观察IVF-ET过程并行胚胎培养。结果:DFI不同的各组患者禁欲时间、精液体积、精子浓度、正常受精率、卵裂率、生化妊娠率、临床妊娠率比较无差异(P0.05)。向前精子活力、正常精子形态率、优质胚胎率、优质胚囊率、囊胚形成率、生化妊娠率、临床妊娠率DFI30%组低于另外两组(P0.05),DFI30%组生化妊娠率、临床妊娠率低于DFI≤15%组(P0.05)。结论:DFI升高会导致精子前向活动力降低,影响精液质量,降低优质胚胎率、优质胚囊率、囊胚形成率,影响妊娠结局。     

三种不同来源精子行卵胞浆内单精子注射临床结局的比较

目的：比较三种方法获得的精子行卵胞浆内单精子注射的临床结局。方法：回顾性分析笔者所在医院2011年1月-2012年12月374个卵胞浆内单精子显微注射（ICSI）治疗周期的妊娠结局,依据精子来源不同分为三组：射出精液组（A组）,经皮附睾穿刺和经皮睾丸穿刺取精子组（B组）,供精组（C组）,比较三组的受精率、卵裂率、优质胚胎率和临床妊娠率。结果：A组的优胚率高于C组（P〈0.017）,三组患者的受精率、卵裂率、临床妊娠率比较差异无统计学意义（P〉0.05）。结论：精液、附睾或睾丸精子、供精行ICSI助孕的结局相近。精子的成熟度、精子的冷冻可能并不影响最终的妊娠率。     

人精子顶体反应检测临床应用价值探讨

[目的]检测Ca2 载体A23187诱导精子顶体反应(AR)率,探讨其临床应用价值。[方法]对51例患者检测A23187诱导精子AR率,分析其与精液常规分析、宫腔内人工授精(IUI)、体外受精(IVF)、单精子卵泡浆内注射(ICSI)结果的关系。[结果]①精液常规分析对诱导AR率的相关性不显著;②诱导AR率低于18.1%理论值时,IVF受精率理论值为0;③3次IUI失败者AR率较受精正常者降低(P<0.05)。④不明原因不育者精子AR率与IVF受精率、卵裂率关系呈正相关(r=0.89,P<0.05),与ICSI受精率、卵裂率无显著关系;用ICSI治疗,其受精率、妊娠率高于IVF治疗组。⑤IVF受精不良病例AR率低,用ICSI治疗后受精率、优质胚胎率升高(P<0.05)。[结论]Ca2 载体A23187诱导顶体反应能预测精子的受精能力,有助于男性不育的临床诊断与治疗。     

SCSA法检测精子DNA碎片指数与辅助生育结局关系的meta分析

目的:探究精子染色质结构分析法(SCSA)检测精子DNA碎片指数(DFI)与体外受精(IVF)、卵胞浆内单精子注射(ICSI)妊娠结局的关系。方法:计算机检索Pubmed、Cochrane library、EMBASE、Springer Link、CNKI、万方和维普等数据库,收集使用SCSA法检测DFI(DFI阈值27%或30%)与IVF、ICSI妊娠结局关系的文献,由两位检索人员根据纳入、排除标准独立筛选文献,提取数据后采用Stata 12.0软件进行meta分析。妊娠结局包括生化妊娠(BP)、临床妊娠(CP)、流产(PL)。相对危险度(RR)和95%可信区间(95%CI)用于评估DFI与妊娠结局的关系。结果:meta分析中纳入9篇有效文献(共2008个周期)。meta分析结果显示,DFI升高显著增加ICSI组生化妊娠率(RR=1.24,95%CI 1.00~1.52)、流产率(RR=2.21,95%CI 1.29~3.78),而与临床妊娠率无显著相关;DFI升高与IVF组的生化妊娠率、临床妊娠率、流产率均无显著相关。结论:SCSA法检测DFI对ICSI的妊娠结局有预测价值,而对IVF的妊娠结局无预测作用,仍需要纳入更多的大样本优质研究以进一步验证。     

不同质量精子对ICSI治疗结局的影响

目的:分析不同质量精子对卵胞浆内单精子注射(ICSI)治疗结局的影响。方法:2000年8月～2007年12月在柳州市妇幼保健院生殖健康助孕中心进行ICSI治疗的352例患者根据精子质量分为睾丸细针穿刺抽吸精子(TESE)组(134例)、严重少弱畸精子组(148例)、精液正常组(70例)。比较3组的受精率、种植率、优胚率、临床妊娠率、流产率、异位妊娠率、分娩率、单胎分娩率、双胎分娩率、早产率和出生缺陷率。结果:3组的受精率、种植率、临床妊娠率、流产率、异位妊娠率、分娩率、单胎分娩率、双胎分娩率、早产率和出生缺陷率比较差异无统计学意义(P>0.05),但在优胚率方面比较差异有统计学意义(P<0.05)。结论:不同质量精子不影响ICSI治疗结局。     

Donor sperm insemination after failed intra-couple intracytoplasmic sperm injection

The purpose of the present multicenter study was to investigate whether an artificial insemination with donor sperm (AID) procedure after intra-couple intracytoplasmic sperm injection (ICSI) failure offers a significant chance of pregnancy and to identify prognostic factors for pregnancy after an AID procedure. An eleven-year retrospective multicenter study was conducted among 13 Centre d’Etude et de Conservation des Oeufs et du Sperme (CECOS) centers. A total of 319 couples having undergone an AID procedure after intra-conjugal ICSI failure were included in this study; a total of 1,159 AID and 1,011 intra-conjugal ICSI cycles were performed. Among the prognostics parameters, the parity and the embryo quality could not be adequately addressed, therefore the parity was not included in the statistical analysis and the embryo quality has been presented as preliminary observations. The pregnancy rate per cycle was 12.0% (139/1,159) and the overall AID pregnancy rate per couple was 43.6% (139/319). Normal or oligoasthenoteratozoospermia (OAT) semen and women aged 34 years or above at the time of AID procedure obtained the lowest AID clinical pregnancy rate. Azoospermia or cryptozoospermia semen and women aged below 34 years obtained the highest AID clinical pregnancy rate. In conclusion, the transition to the AID procedure after intra-conjugal ICSI failure allows such couples to obtain a pregnancy, however after each ART failure AID transition should be proposed according to the woman’s age and sperm characteristics.

Abbreviations: AID: artificial insemination with donor sperm; ICSI: intracytoplasmic sperm injection; CECOS: Centre d’Etude et de Conservation des Oeufs et du Sperme; OAT: oligoasthenoteratozoospermia; IVF: in vitro fertilization; ART: artificial reproductive technology; β hCG: beta human chorionic gonadotrophin; SD: standard deviation; OR: Odds ratio     

积水输卵管液对精子运动及形态学的影响

目的:通过体外研究积水输卵管液对精子运动及形态学的影响,探讨输卵管积水引起不孕的原因,并为单侧输卵管积水(对侧输卵管通畅)患者的处理提供依据。方法:优化处理后的高质量的精子体外与输卵管积水患者的输卵管液(积水组n=18)及正常输卵管液(对照组n=18)孵育2h、7h和24h后,精子分析仪(CASA)测量精子的各项运动参数,应用精子Diff-Quick涂片染色法分析精子形态。结果:①培养2h、7h和24h后,各时间点积水组精子的前向性运动百分率、平均曲线运动速度(VCL)、平均直线运动速度(VSL)、平均路径运动速度(VAP)、快速直线运动精子活率(MOT)、直线性(LIN)、摆动性(WOB)及前向性(STR)各项精子运动参数均低于对照组(P<0.05),且输卵管积水加快精子的运动速度参数(VCL、VSL、VAP)随时间延长而降低的速度(P<0.05)。②培养2h、7h和24h后,积水组与对照组精子的正常形态率对比无统计学差异,且不随时间延长而改变(P>0.05)。结论:输卵管积水在体外能降低精子的前向性运动百分率、精子运动速度参数(VSL、VCL、VAP)及精子运动方式参数(MOT、LIN、WOB、STR)各项运动指标,并提示输卵管积水不仅干扰了精子活动力,并可能对精子的获能产生不良影响,推测体内输卵管积水对精子产生一定的损害作用,从而影响受孕,但对精子正常形态精子率未有明显影响。     

Numerical chromosome anomalies in human sperm at low sperm concentration

INTRODUCTION: With the assisted reproduction techniques the natural selection of sperm is bypassed on different levels. AIM: The aim of the present work is to determine the frequencies of sperm numerical chromosome aberrations in infertile men with low sperm count and to examine the relationship between the spermatogram parameters (sperm count and motility) and the aneuploidy and diploidy frequencies. METHODS: 32 men with low sperm count were investigated. Semen analysis was performed according to the WHO criteria. Disomy and diploidy frequencies were detected with fluorescence in situ hibridization using 17, X and Y centromeric probes. 200,969 sperm were scored, with the mean of 6272 cells in each subjects. The rate of numerical chromosome anomalies were also estimated using the detected disomy and the diploidy frequencies. RESULTS: Mean sperm concentration of the 32 men was 18.2 million/ml (SD: +/- 8.43, range: 8.0-45.5), mean motility 49.4% (+/- 9.32, 30-69.2). The X/Y ratio was 1.07. The mean frequencies of sex chromosome disomy, chromosome 17 disomy and diploidy were 0.36%, 0.16% and 0.56%, respectively. The most frequent disomy was XY disomy (0.14). Sex chromosome disomy frequencies were higher in oligospermic samples (0.37% vs. 0.32%, OR = 1.18, 95% CI = 1.01-1.39, p < 0.001). With special regard to XY disomy (0.08% vs. 0.17%, OR = 1.99, 95% CI = 1.48-2.67, p < 0.001). In subjects with oligoasthenozoospermia the diploidy frequency increased (0.96%, OR = 2.39, 95% CI = 2.13-2.69, p < 0.001), mostly due to the elevated rate of diploid sperm of meiosis II. origin (XX or YY diploids). Estimated frequency of the numerical chromosome anomalies was 8.3 +/- 5.3 in the study population. Neither the sperm count, nor the sperm motility showed correlation with the detected frequency of the chromosome aberrations. CONCLUSIONS: In oligospermic patients there is a risk for elevated frequency of sperm with sex chromosome aneuploidy, especially the XY disomy. Furthermore, the diploidy frequency is increased in oligozooastenospermic samples. Nevertheless, classical parameters of semen analysis (sperm count and motility) do not correlate with the frequency of numerical chromosomal anomalies. The risk can be determined using fluorescens in situ hybridization on sperm.     

形态正常精子百分率与顶体酶活性的关系

目的:探讨精子形态与精子顶体酶活性的关系。方法:对737例男性不育患者采用分光光度比色法测定顶体酶活性,采用精子形态检测系统下人工修正方法分析精子形态。结果:精子形态正常组的顶体酶活性明显高于精子形态异常组,两组间比较差异显著(P<0.05);顶体酶活性正常组形态正常精子百分率明显高于顶体酶活性异常组形态正常精子百分率,两组间比较差异亦具有显著性(P<0.001);顶体酶活性与形态正常精子百分率间呈显著正相关关系(r=0.172,P<0.001)。结论:精子形态可影响精子顶体酶活性。     

精子核成熟度与精子形态的关系

目的:研究精子核成熟度与精子形态关系,探讨精子形态异常机制。方法:收集55例精液样本,其中生育组19例,畸形精子症组36例。应用CASA分析精子密度、活力,伊红染色检测精子活率,联苯胺染色分析精液白细胞浓度,改良巴氏染色分析精子形态,苯胺蓝染色法评价精子核成熟度。结果:畸形精子症组苯胺蓝染色阳性率显著高于生育组(P<0.05)。头部异常、颈部异常、尾部异常、无定型、其它畸形精子组苯胺蓝染色阳性率显著高于形态正常精子组苯胺蓝染色阳性率(P<0.05)。结论:精子核成熟异常可导致精子形态异常。     

不同来源精子行卵胞浆内单精子显微注射结局分析

目的 探讨不同来源精子行卵胞浆内单精子显微注射后的妊娠结局.方法 回顾分析218个卵胞浆内单精子显微注射周期,按精子来源分为射出精子组190个周期,附睾穿刺取精组12个周期,睾丸穿刺取精组16个周期,比较3组的受精率、卵裂率、优质胚胎率和临床妊娠率等指标.结果 睾丸穿刺取精组较射出精子组及附睾穿刺取精组的受精率、卵裂率高,经比较差异均有统计学意义(χ2值分别为15.27、10.83、7.87、4.25,均P<0.05);睾丸穿刺取精组与射出精子组及附睾穿刺取精组临床妊娠率比较差异均无统计学意义(χ2值分别为1.67、0.94,均P>0.05);3组间的流产率、畸形率均为0,差异均无统计学意义(均P>0.05).结论 附睾穿刺取精组及睾丸穿刺取精组来源精子行卵胞浆内单精子显微注射助孕可获得与射出精子组相似的临床妊娠率.     

Effects of bull, sperm type and sperm pretreatment on male pronuclear formation after intracytoplasmic sperm injection in cattle

This study investigated the effects of the bull, sperm type (dead, immotile or motile) and sperm pretreatment (i.e. mechanical (tail-cutting or tail-scoring) or chemical (heparin, heparin + caffeine, calcium ionophore A23187 or dithiothreitol)) on male pronuclear formation after intracytoplasmic sperm injection (ICSI) in cattle. Three experiments were conducted. In Experiment 1, spermatozoa from three bulls (A, B and C) were used for both ICSI and in vitro fertilization (IVF). The results were that sperm from bull B yielded a higher penetration/male pronuclear formation rate than that of bull C when used for IVF (89.6% v 25.6%, P<0.01). However, when injected into oocytes by ICSI, sperm from bull C had a higher male pronuclear formation rate than that of bull B (34.6% v. 16.1%, P<0.05). The effects of sperm type and mechanical pretreatment were examined in Experiment 2. No significant difference was found in the male pronuclear formation rate when the three types of sperm were injected into oocytes. Tail-scored sperm achieved a higher male pronuclear rate than that of non-mechanically treated ones (38.2% v. 13.2%, P<0.005). In Experiment 3, chemical pretreatments were tested and compared. Higher male pronuclear rates, compared with the control, were obtained when sperm were pretreated with heparin + caffeine, calcium ionophore A23187 and dithiothreitol (48.2%, 62.5% and 64.5% v. 25.0%, P<0.05, 0.005 and 0.005, respectively). These results indicate that (1) there is a bull variation in male pronuclear formation with ICSI, and the male pronuclear rate by ICSI is not coincident with the results by IVF, (2) immobilization of a spermatozoon by tail-scoring before ICSI can improve the formation of the male pronucleus, and (3) an appropriate chemical pretreatment of spermatozoa is necessary to achieve a higher rate of male pronuclear formation.     

睾丸新鲜精子与冷冻复苏精子临床妊娠结局的比较

目的 比较无精子症患者睾丸穿刺取精(testicular aspiration, TESA)与睾丸显微取精(microdissection testicular speron extraction, m-TESE)新鲜与冷冻复苏精子进行卵胞质内单精子注射(intracytoplasmic sperm injection, ICSI)治疗的结局,以及新鲜、冷冻复苏胚胎移植的妊娠结局。方法 选取2019年6月至2021年3月在华中科技大学同济医学院生殖医学中心接受ICSI助孕的1 116个周期,常规超排卵和经阴道B超引导下取成熟卵子进行ICSI,梗阻性无精子症(obstructive azoospermia, OA)患者行TESA,非梗阻性无精子症(nonobstructive azoospermia, NOA)患者行m-TESE,比较TESA新鲜精子组和冷冻复苏精子组,m-TESE新鲜精子组和冷冻复苏精子组的受精率、卵裂率、优质胚胎率、临床妊娠率、流产率和活产率,以及OA、NOA患者新鲜和复苏胚胎移植的临床妊娠率、流产率和活产率。结果 在受精率方面,TESA新鲜精子组明显高于TESA冷冻...     

吸烟对精子DNA完整性、精子参数的影响

目的:探讨吸烟对精子DNA完整性、精子参数的影响。方法:调查191例男性不育患者的吸烟情况,采用吖啶橙荧光染色后检测精子DNA完整性,计算机自动分析精子密度与活力,精子形态检测系统下人工修正方法进行精子形态分析。结果:吸烟组精子活力显著低于不吸烟组(P<0.05),精子密度和形态低于不吸烟组,但差异无统计学意义(P>0.05)。吸烟组精子DNA完整率异常例数显著高于不吸烟组(χ2=5.393,P<0.05),精子DNA完整率异常组精子密度显著低于精子DNA完整率正常组(P<0.05),精子DNA完整率异常组的精子活力、形态与正常组相比差异无统计学意义(P>0.05)。结论:吸烟影响精子DNA完整率;精子DNA完整率异常与精子参数异常相关;吸烟可能通过影响精子DNA完整性影响精子参数。