不同参数精子顶体酶活性与精子在IVF治疗中受精能力的关系研究

目的探讨不同参数精子的顶体酶活性与精子在体外受精(IVF)治疗中受精能力的关系。方法通过对75例接受IVF或短时IVF治疗的少弱精子或正常精子病例进行术前精子顶体酶活性测定,不明原因和有受精不良史病例优先入组。比较顶体酶正常值组和低值组的精子参数差异和IVF受精结果;进一步比较精子高浓度组(35×10~6/m L)和低浓度组[(10～35)×10~6/m L]的顶体酶活性值差异和IVF受精结果。结果顶体酶活性正常值组和低值组的精子浓度分别为(61.88±35.95)×10~6/m L和(32.89±12.94)×10~6/m L,两者差异具统计学意义(P0.01),前向运动精子百分率分别为(48.76±12.60)%和(36.35±10.30)%,差异无统计学意义(P0.05),快速前向运动精子百分率分别为(22.41±7.02)%和(16.35±5.23)%,差异无统计学意义(P0.05), IVF或短时IVF总受精率分别为67.65%和41.74%,两者差异具统计学意义(P0.01),不受精率分别为19.51%和38.24%,差异无统计学意义(P0.05),低受精率0%VS 14.71%(P0.05)。精子高浓度组和低浓度组的精子顶体酶活性值分别为(76.55±32.65) VS (43.17±19.68)(P0.01), IVF或短时IVF总受精率70.65%VS 41.20%(P0.01),不受精比例17.95%(7/39) VS 38.89%(14/36)(P0.05),低受精比例0%(0/39)VS 13.89%(5/36)(P0.05)。精子高浓度组完全不受精病例卵子数均大于6枚,没有低受精病例,顶体酶异常占12.8%(5/39),其中只有1例不受精;而精子低浓度组完全不受精病例大部分卵子数少于6枚,顶体酶异常占80.56%(29/36),其中不受精和低受精率为58.62%(17/29)。结论在IVF治疗中,粗子浓度与受精能力的相关性比活动力和顶体酶活性更强;在低浓度组通常伴有顶体酶活性的低下,提示低受精或不受精可能性增大;而高浓度精子顶体酶大多正常,呈现"全"或"无"的受精模式,其不受精原因可能与顶体酶活性无关。     

精子CMA3阳性率与IVF受精率的相关性研究

目的探讨精子色素酶A3(CMA3)阳性率与IVF受精率之间的相关性。方法收集2015年4月至2016年7月因单纯输卵管性不孕在我院生殖医学科行常规IVF助孕的156个周期,以受精率<25%为低受精,根据受精率不同分为正常受精组(134个周期)和低受精组(22个周期)。取卵日,将授精后剩余的洗涤精液行精子浓度和活动率分析,并将精子进行CMA3染色。比较两组优选后的精子参数,分析IVF受精率与精子参数及CMA3阳性率的关系。结果与正常受精组比较,低受精组精子CMA3阳性率显著升高[(20.0±4.2)%vs.(30.7±2.3)%],优选后前向运动精子的百分率显著降低[(90.4±4.8)%vs.(74.3±3.4)%](P<0.05);两组精子浓度比较无显著性差异(P>0.05)。IVF受精率与优选后的前向运动精子百分率呈正相关(r=0.76,P<0.01)。精子CMA3阳性率与优选后前向运动精子百分率(r=-0.82,P<0.01)及IVF受精率(r=-0.83,P<0.01)呈显著负相关,与精子浓度则无显著相关性(P>0.05)。结论精子CMA3阳性率与IVF受精率负相关,但其具体机制尚需进一步研究探讨。     

精子顶体酶检测对不明原因不孕夫妇助孕治疗方案选择的临床意义

目的:探讨精子顶体酶检测对不明原因不孕(UI)夫妇选择助孕治疗方案的临床意义。方法:回顾性分析2013年1月至2015年12月诊断为UI,并经3次宫腔内人工授精(IUI)治疗失败改行体外受精-胚胎移植(IVF-ET)助孕的49对夫妇共49个周期的临床资料,并对比分析同期因输卵管阻塞因素行常规IVF治疗的95对夫妇的131个周期的临床资料,比较两组实验室数据、临床结局和顶体酶活性差异;进一步根据UI夫妇男方精子顶体酶活性将UI组分成2个亚组:36 IU/106精子组共20个周期,≥36 IU/106精子组共29个周期,比较两亚组之间受精率的差异。结果:1行IVF-ET治疗的UI夫妇,比同期因输卵管阻塞行常规IVF的夫妇受精率显著下降(67.0%vs 76.4%,P0.05),补救性卵细胞胞质内单精子注射(ICSI)率高于输卵管因素组(20.4%vs 6.1%,P0.05);成熟卵母细胞比例(MII卵率)、可利用胚胎率、优胚率、种植率、临床妊娠率两组差异无统计学意义(P0.05)。2UI组中位精子顶体酶活性低于输卵管因素组(36.03 IU/106精子vs 61.98 IU/106精子,P0.01),UI组中顶体酶活性36 IU/106精子亚组的受精率明显低于顶体酶活性≥36 IU/106精子亚组的受精率(47.7%vs80.3%,P0.01)。结论:1UI夫妇男方精子顶体酶活性低下导致的受精率低,可能是造成女性不孕的主要原因和潜在因素。2鉴于UI夫妇男方精子顶体酶活性36 IU/106精子时受精率较低,建议不采用IUI治疗,可选择IVF+短时受精联合早期补救ICSI治疗。     

顶体酶活性对体外受精-胚胎移植结局的影响

目的:探讨精子顶体酶活性对体外受精-胚胎移植(IVF-ET)结局的影响。方法:选择909对在本院生殖中心就诊的不孕夫妇,其中女方检查为正常或仅为输卵管因素不孕。采用改良巴氏法染色和Na-苯甲酰-DL-精胺酸-P-硝酰基苯胺(BAPNA)法分别对丈夫的精液进行精子形态学分析和精子顶体酶活性测定。结果:顶体酶异常组中各项精液参数,包括正常形态精子百分率、精子活动率、活动力、快速前向运动精子百分率及精子密度均显著低于顶体酶正常组(P<0.01);顶体酶活性与精液常规分析上述各参数之间存在非常显著的正相关(P<0.01);两组间取卵数、卵裂率、优质胚胎率、胚胎冷冻率、无可移植胚胎周期百分率、胚胎移植数和流产率均无显著性差异(P>0.05);顶体酶正常组的受精率、仅一个胚胎移植的周期百分率、着床率及临床妊娠率均明显高于顶体酶异常组(P<0.01)。结论:顶体酶活性异常与精液常规分析各主要参数的异常密切相关,可导致体外受精率显著下降。顶体酶活性检测在IVF结局预测中起着重要作用。     

精子形态率、顶体酶活性对IVF受精失败的影响

目的 探讨精液中精子形态率及顶体酶活性与IVF受精失败的关系.方法 回顾性分析2014年在本中心行IVF新鲜周期的982个周期患者,根据精子形态分析结果,将正常精子形态百分率(x)分为5组,包括:1组:x≥4％,2组:x＜1％,3组:1％≤x＜2％,4组:2％≤x＜3％,5组:3％≤x＜4％.根据顶体酶活性分为正常组及异常组.结果 IVF受精失败组(受精率＜30％)与正常组比较,正常精子形态率及顶体酶活性差异有统计学意义(P＜0.01).精子形态率比较,2、3、4组与1组比较,差异有统计学意义(P＜0.01).5组与1组比较,差异无明显统计学意义(P＞0.05).顶体酶活性正常组与异常组比较,受精失败率差异有统计学意义(P＜0.01).结论 精子形态率及顶体酶活性异常可导致IVF受精失败率升高.     

Percoll优选对形态正常精子百分率和顶体酶活性的影响

目的 :了解形态正常精子百分率和顶体酶活性在Percoll优选前后的变化。　方法 :30例不育男性精液标本 ,应用计算机精液分析仪进行精子形态分析 ,分光光度比色法检测顶体酶活性。　结果 :Percoll优选后形态正常精子百分率和精子顶体酶活性均明显高于Percoll优选前 (P均 <0 .0 0 1)。　结论 :Percoll法优选精子后形态正常精子百分率和顶体酶活性增加。     

精索静脉曲张患者精子顶体酶活性研究

目的　探讨精索静脉曲张患者在精索静脉高位结扎术前、后的精液常规和精子顶体酶活性 (SAA)的变化 ,以及它们之间的相关性。方法　分别用常规方法和改良的Kennedy’s法测定精液常规和精子顶体酶活性 ,再分析它们之间的相关性。结果　精索静脉高位结扎术后患者的精子密度、形态正常率、存活率、活动力以及精子顶体酶活性都较术前明显增高 (P <0 .0 5 ) ;术前、后精子顶体酶活性的变化与精子密度的变化和与精子形态正常率的变化呈正相关关系 (P <0 .0 5 )。结论　精索静脉高位结扎术后精索静脉曲张患者精液质量如精子密度、形态正常率、存活率、活动力和精子顶体酶活性得到明显的提高 ,而且术后精子顶体酶活性的提高与精子密度和形态正常率的提高具有正相关关系     

精子功能指标在辅助生殖助孕方式选择中的作用

目的评估精子功能参数对精子常规体外受精能力的预测价值,探寻有效选择受精方式的新指标。方法回顾性分析2017年1月至2019年12月于我院首次行体外受精-胚胎移植(IVF-ET)短时受精的429个周期的临床资料,根据受精结局不同分为2组:受精失败或受精率<30%的患者为早补救卵胞浆内单精子注射(R-ICSI)组(n=98);受精率≥30%的为短时IVF组(n=331)。比较两组患者精子前向运动(PR)、精子浓度、精子正常形态率、顶体酶活性、顶体反应(AR)、酪氨酸磷酸化(TP)及透明质酸结合试验(HBA)等指标;采用Logistic回归分析影响受精的独立危险因素,并绘制受试者工作曲线(ROC曲线)评估各项危险因素对IVF受精率的预测价值。结果两组患者精子的正常形态率、顶体酶活性、自发AR和TP比较均无显著性差异(P>0.05);R-ICSI组患者的精子浓度、PR、诱发顶体反应(IAR)和HBA均显著低于短时IVF组(P<0.05)。Logistic回归分析显示,精子PR、IAR和HBA为IVF受精率的独立危险因素;ROC曲线显示精子PR、IAR、HBA和联合预测新变量(Pre-1)的曲线下面积(AUC)分别为0.66、0.62、0.64和0.70。结论精子功能指标中IAR和HBA是影响IVF受精率的独立危险因素,其对辅助生殖助孕方式的选择有一定指导意义。     

精液白细胞对精子质量及IVF受精率的影响

目的探讨精液中白细胞对精子质量以及对常规体外受精率的影响。方法分析35例门诊白细胞精子症患者及对照组的精子密度、活力、前向运动精子比例及精子畸形率。分析28例常规体外受精(IVF)白细胞精子症及其对照组的受精率。结果白细胞精子症患者精子密度、活力、前向运动精子比例均低于对照组,精子畸形率高于对照组,IVF受精率低于对照组。结论精液白细胞是可能造成精子质量下降,IVF受精率降低的原因之一,纠正白细胞精子症对于治疗男性不育,提高IVF受精率有重要意义。     

精子形态可否预测精子其他参数和受精结局

目的探讨精子形态在辅助生殖治疗中对选择授精方法时有否预测价值和意义。方法回顾性分析2012年6月至2015年2月接受辅助生殖治疗的1 951周期患者精液资料,根据《WHO人类精液检查与处理实验室手册》(第五版)精子正常形态率参考值下限为4%,将患者的精子正常形态率分成3组:A组1.0%470周期,B组1%~4%562周期,C组≥4%919周期,分别采用常规体外受精(IVF)或卵胞浆内单精子注射(ICSI)以及IVF受精失败后补救ICSI(RICSI)授精方式,比较3组精子正常形态与其他精液参数相关性,与授精方式、受精率、优质胚胎率的关系。结果除了顶体酶外,精子浓度、活力、前向运动率、DNA完整率在3组形态间都差异极显著(P0.01),而且它们与精子正常形态呈显著的正相关;授精方式上3组的IVF比例分别是72.3%、86.7%、96.8%,ICSI组27.7%、13.3%、3.2%,3组间授精方式差异极显著(P0.01),RICSI比例也随着精子正常形态率升高而降低;受精结局上,只有IVF组3组形态间受精率(67.0%、70.0%和79.6%)差异极显著(P0.01),且和精子正常形态率趋势一致;胚胎质量反映在不同授精方式上趋势各有不同。结论精子正常形态与其他大部分的精液参数有正相关性,影响IVF的受精率,故在辅助生殖治疗中对授精方式选择有一定指导意义。     

Correlation between sperm morphology, acrosin, and fertilization in an IVF program

Acrosin, a neutral proteinase, is located within the acrosome. The aim of this study was to evaluate acrosin concentrations in patients with severe damage of the sperm head and to determine whether acrosin concentration could predict the chances of fertilization in an IVF program. Sixty patients were accepted into this study, prospectively. The patients were divided into two groups, those with a normal morphology of less than 14% (group I, n = 33) and those with normal morphology less than 14% (group II, n = 27). All patients had a sperm concentration of less than 20 million sperm/ml and less than 30% progressively motile sperm. The acrosin assays were performed on the semen sample obtained on the day of IVF. Routine IVF insemination procedures were used, and only mature oocytes were considered. The only factor that showed a significant correlation of fertilization was normal morphology (p less that 0.01). The mean acrosin level was 73.4 /+- 38.6 mED/10 million sperm in group I and 70.9 /+- 42.7 mIU/10 million sperm in group II (no significant difference). The fertilization rate in group I was 45.4% and in group II, 77.7% p less than 0.002). Acrosin levels were not significantly different in patients with and without fertilization (72.0 /+- 42.1 and 73.6 /+- mIU/10 million sperm, respectively).     

Changes in human sperm chromatin stability during preparation for in-vitro fertilization

This study was designed to define the effects of sperm preparation on sperm chromatin stability in relation to in-vitro fertilization (IVF). Semen samples used for IVF-embryo transfer (ET) in the treatment of infertility due to tubal factors were studied. Cases with semen variables below reference limits in previous samples were excluded. Sperm were prepared by a swim-up technique employing either of two different tissue culture media, Ham's F-10 or Earle's balanced salt solution. Sperm chromatin stability was tested by exposure both to sodium dodecyl sulphate (SDS) only and SDS together with a zinc-chelating agent, disodium ethylene diamine tetraacetate (SDS-EDTA). Sperm head swell scores were defined under different experimental conditions and the relationship to sperm motility, morphology, fertilization rate and pregnancy occurrence was tested. No differences were seen between the chromatin stability of sperm from the original sample and that after swim-up preparation, neither immediately after completion of the swim-up procedure, nor at the time of insemination of ova. With time, the chromatin became more stable, which occurred to a similar extent both in the original sample and in swim-up preparations using Ham's F-10. Otherwise, sperm chromatin stability was unaffected by either of the two media used for swim-up. At higher incubation temperatures, decondensation in SDS was enhanced. Altogether, no correlation was found between sperm chromatin stability or enhancement of decondensation by temperature and the success of IVF treatment expressed in fertilization rates or pregnancies. The results are reassuring in that only small changes in sperm chromatin stability occurred during the preparation for IVF. As long as semen of presumably good quality is used, these changes in chromatin stability do not seem to be of clinical importance.     

Seminal reactive oxygen species as predictors of fertilization, embryo quality and pregnancy rates after conventional in vitro fertilization and intracytoplasmic sperm injection

High seminal reactive oxygen species (ROS) are related to poor semen quality and impaired fertilization. We aimed at finding whether there is an association between ROS and fertilization, embryo quality and pregnancy rates after conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). In prepared semen of 147 male partners of infertile couples, ROS were assessed with luminol chemiluminescence. Spermiogram was assessed in native semen. ROS were negatively correlated with standard sperm characteristics and testicular volume, and positively with abnormal sperm head morphology. Fertilization rate and embryo morphology on day 2 and on day 4 were assessed in 41 IVF and 106 ICSI cycles. The influence of maternal (female age and number of oocytes) and paternal (sperm motility, morphology and ROS) factors on fertilization and embryo quality were assessed by means of regression analyses. After IVF, fertilization and pregnancy rates were negatively associated with ROS level (p = 0.031 and 0.041, respectively). In case of higher ROS, significantly fewer ICSI-derived embryos (p = 0.036) reached the morula-blastocyst stage on day 4. High seminal ROS levels are associated with impaired sperm fertilizing ability and lower pregnancy rates after IVF. In ICSI, a negative association of ROS with embryo development to the blastocyst stage has been observed.     

精子形态对体外受精的影响作用

目的　探讨精子形态在体外受精-胚胎移植（IVF-ET）中对受精率及胚胎卵裂速度、胚胎形态的影响作用。　方法　观察89个IVF-ET周期精子处理前后各项参数及成熟卵的受精率、卵裂胚胎的细胞数及形态,分析不同受精密度下精子形态对受精率及胚胎发育的影响。　结果　受精密度≤1.2×10     

Human sperm morphology and in vitro fertilization: sperm tail defects are prognostic for fertilization failure

Summary. The aim of this study was to determine the relationship between sperm morphology and fertilization rates in vitro. Semen samples were obtained from 50 couples undergoing IVF treatment. Sperm morphology was classified by strict criteria (Tygerberg) according to head, midpiece and tail defects in neat semen and after sperm selection by Percoll gradient centrifugation. Percoll preparation significantly increased the percentage of sperm with normal morphology from 13 to 20%. However, the greatest single regression coefficient was observed with the percentage of sperm with tail defects and correlated negatively with fertilization rates in vitro both before and after Percoll preparation. Therefore, tail morphology may be of value as a prognostic factor in assisted conception both before and after Percoll preparation.—     

Does computerized image analysis of sperm movement enhance the predictive value of semen analysis for in-vitro fertilization results?

The influence of semen quality on fertilization rates in an in-vitro fertilization (IVF) programme was studied by analysing both conventional semen parameters and computerized movement characteristics. The study was based on 407 inseminated oocytes which were obtained from 50 patients in 113 laparoscopies. Sperm concentration did not correlate strongly with the fertilization rate. Sperm motility and morphology were the most meaningful parameters in predicting fertilization success. A drop in fertilization rate was found when sperm motility or normal morphology were below 40%. Sperm velocity measured in semen was the only sperm movement parameter which correlated with the fertilization rate, albeit weakly. The latter was reduced when average sperm velocity in semen was less than 50 microns/sec. Conventional semen parameters seem to be more predictive of the fertilizing potential of an ejaculate than movement characteristics obtained by computerized image analysis.