ICSI中不同来源精子对临床结局的影响

目的:分析不同来源精子在卵细胞胞质内单精子注射(ICSI)后的临床结局。方法:将682个ICSI治疗周期分为:射出精子组(598例)、经皮附睾精子抽吸术(PESA)得到精子组(58例)、睾丸精子获取术(TSE)得到精子组(26例),比较三组的受精率、临床妊娠率、种植率、流产率、异位妊娠率以及分娩率之间的差别。结果:TSE组受精率明显低于射出精子组和PESA组(81.06%vs87.95%,87.82%,P<0.05);射出精子组、PESA组和TSE组的妊娠率(39.46%,48.28%,34.62%)、种植率(19.80%,23.80%,18.34%)、流产率(13.13%,17.86%,11.11%)、异位妊娠率(5.51%,7.14%,11.11%)、分娩率(32.11%,36.21%,26.92%),均没有显著差异(P>0.05)。结论:虽然TSE来源精子对ICSI受精率有所影响,但射出精子和手术(TSE和PESA)来源精子对妊娠结局没有影响。     

卵胞浆内单精子注射治疗严重男性因素不育症428个周期的临床分析

目的 分析卵胞浆内单精子显微注射技术(intracytoplasmic sperm injection,ICSI)在治疗严重男性因素不育症时的有效性和安全性. 方法 回顾分析2009年1月至2010年12月间在本院因严重男性不育症行ICSI助孕的428例患者的数据,其中245例梗阻性无精子症组采用经皮附睾穿刺抽吸术取得的附睾精子;183例严重少弱畸精子症组采用的是射出精液中的精子.并且收集患者的年龄、体重指数、基础卵泡刺激素(FSH)、移植胚胎数目、受精率、优胚率、取消率、妊娠率、着床率、流产率、宫外孕发生率、妊娠出生率、移植出生率、出生异常率等资料. 结果 梗阻性无精子症组受精率(84.1±1 7.1)％,着床率45.2％,移植出生率59.1％,流产率10.3％,出生异常率3.5％.严重少弱畸精子症组受精率(79.9±20.9)％,着床率37.8％,移植出生率48.2％,流产率15.2％,出生异常率3.1％.两组均获得了满意的临床结局,但两组的受精率、着床率、移植出生率及流产率均存在统计学差异. 结论 本研究结果提示梗阻性无精子组的受精率,着床率及移植出生率较严重少弱畸精子症组均明显升高,且术后流产率较低,提示梗阻性无精子症患者经附睾穿刺行ICSI较严重少弱畸精子症患者可以获得更好的临床结局.但此技术的安全性,依然需要更多的研究.     

体外受精-胚胎移植后三胎妊娠减胎术结局分析

目的分析早期选择性减胎术对妊娠并发症和新生儿的影响,评价其安全性。方法收集本中心2000年1月至2003年2月经体外受精胚胎移植(IVF ET)和卵胞浆内单精子注射(ICSI)受孕的三胎妊娠减灭为双胎妊娠35例(A组)及同期双胎妊娠166例(B组)。比较两组妊娠期并发症、剖宫产率、围产期死亡率、新生儿畸形率、平均出生体重及孕周。结果两组自然流产率、妊娠高血压综合征发生率、分娩率、剖宫产率、早产率、围产期死亡率、畸形率、平均出生孕周差异均无显著性;胎膜早破率和先兆早产率A组分别为30%、7.7%,B组分别为13%、56%,两组比较差异有显著性(P<0.05);平均出生体重A组(2410.8±570.0)g,明显低于B组(2560.9±530.6)g(P<0.05)。结论三胎妊娠早期选择性减胎术后胎膜早破和先兆早产率明显升高,胎儿存在低出生体重可能性。因此,预防多胎妊娠,限制胚胎移植数目,仍然是辅助生育技术实施过程中不可忽视的问题。     

经皮附睾穿刺取精精子冷冻复苏后卵细胞胞质内单精子注射治疗无精子症的初步研究

目的:回顾性分析27例无精子症患者经皮附睾穿刺取精术(PESA)所获精子冷冻复苏后行卵细胞胞质内单精子注射(ICSI)治疗后的效果及妊娠结局。方法:将诊断性附睾穿刺以及PESA治疗周期ICSI后所剩余活精子以常规方法加以冷冻,将复苏后找到了足量活精子并行ICSI的病例归为冻精组,而采用新鲜PESA活精子ICSI的病例则归为对照组。比较冻精组与对照组的受精率、种植率、临床妊娠率,同时分析两组间的妊娠并发症、新生儿出生及畸形等情况。结果:冻精组15个周期、对照组100个周期分别注射MⅡ期成熟卵子163、1 157个,受精率冻精组显著高于对照组(84.05%vs73.29%,P<0.05),种植率、临床妊娠率则两组间差异无显著性(23.07%vs15.73%;53.33%vs37.00%,P>0.05),新生儿出生体重差异亦无显著性(P>0.05)。冻精组共妊娠8例,已分娩5例,继续妊娠3例。对照组妊娠37例,已分娩30例,1例死胎;继续妊娠3例;流产4例。两组均未出现重大的妊娠并发症及新生儿畸形。结论:采用PESA冷冻精子ICSI是治疗男性无精子症的一种经济、有效、安全的方法;但PESA冻精复苏率有待于进一步提高。     

睾丸精子行ICSI改善严重畸形精子症患者治疗结局5例报告

目的:探讨利用睾丸精子行卵细胞胞质内单精子注射(ICSI)治疗严重畸形精子症患者(精液或附睾液精子畸形率≥99%)的可行性,改善辅助生殖技术治疗结局。方法:回顾性分析5例严重畸形症精子患者(附睾液精子,n=4;精液精子,n=1)利用不同来源精子行ICSI治疗的临床资料,并比较睾丸精子组与非睾丸精子组(附睾液精子和精液精子)之间受精率、卵裂率、优质胚胎率、妊娠率以及种植率的差异。结果:5例严重畸形精子症患者取精液精子或附睾液精子行ICSI治疗后无1例妊娠,而改用睾丸精子行ICSI治疗后4例成功妊娠。睾丸精子组与非睾丸精子组之间受精率、卵裂率及优质胚胎率均无显著差异(P>0.05),而睾丸精子组妊娠率和种植率均显著高于非睾丸精子组(P<0.01)。结论:对应用附睾精子或精液精子行ICSI治疗失败的严重畸形精子症患者改用睾丸精子治疗可有效改善其治疗结局。     

不同源性精子经ICSI-ET助孕术后的临床结局及子代安全性研究

目的探讨附睾/睾丸穿刺取精精子及射出精子经ICSI-ET助孕术后临床妊娠结局及子代安全性有无差别。方法采用回顾性队列研究,选择2017年1月至2019年5月行ICSI-ET助孕并新鲜胚胎移植的1248个周期为研究对象,根据男方取精方式的不同分为两组,即附睾/睾丸穿刺取精组(n=230)和对照组(n=1018),比较分析两组患者的一般情况、促排卵实验室指标、临床结局及围产期结局指标。结果手术取精组患者平均获卵数[(12.62±4.19)(11.74±4.03)]及MⅡ卵数[(10.70±4.17)(9.79±3.83)]相比对照组较多,但是ICSI后的受精率(77.02%79.28%)及正常受精率(72.16%74.56%)低于对照组,P值均0.05。手术取精组平均临床妊娠率高于对照组(65.22%57.86%),有统计学差异(P0.05);但是两组间的多胎率、早期流产率、胎儿畸形引产率、活产率、剖宫产率、37周早产率及双胎活产的构成比均无统计学差异(P0.05),单胎活产或双胎活产的子代出生孕周和出生体重均无统计学差异(P0.05)。结论不同源性的精子经ICSI-ET助孕后主要影响受精率,手术取精精子经ICSI-ET后临床妊娠率优于射精精子,围产期结局及子代安全性并无差异。     

睾丸精子冷冻复苏对ICSI助孕结局的影响

目的探讨睾丸精子冷冻复苏对ICSI助孕结局的影响。方法回顾性分析2010年1月至2014年9月因梗阻性无精子症在本院采用睾丸精子行ICSI助孕的214个周期的临床资料,其中107个周期采用冷冻复苏的睾丸精子(冻精复苏组),另外107个周期采用新鲜睾丸精子(新鲜精子组)。比较两组中女方的一般资料、受精率、卵裂率、可利用胚胎数、优质胚胎率,以及临床妊娠率。结果本研究中,107个周期冻存睾丸精子复苏均获成功;冻精复苏组与新鲜精子组行ICSI助孕后的受精率[(76.91±18.24)%vs.(75.35±20.62)%]、卵裂率[(94.69±5.29)%vs.(95.37±4.48)%]、可利用胚胎数[(4.67±2.52)vs.(4.20±2.75)个]、优质胚胎率[(64.47±26.08)%vs.(60.34±27.39)%]及临床妊娠率(52.24%vs.50.63%)比较均无显著性差异(P0.05)。结论睾丸精子冷冻复苏对ICSI助孕结局并无显著影响。     

精子DNA完整性对冻融胚胎移植周期妊娠结局的影响

目的探讨男性精子DNA完整性对行ART助孕患者冻融胚胎移植(FET)周期妊娠结局的影响。方法回顾性分析2016年8月至2017年12月在江苏省苏北人民医院生殖中心接受IVF/ICSI助孕的667对不孕夫妇FET周期的临床资料。男性患者进入周期前均采用精子染色质结构分析法(SCSA)染色后通过流式细胞仪来检测男性精子DNA完整性,并观察精子DNA碎片率(DFI)对妊娠结局的影响。根据不同精子DFI水平将研究对象分为两组:低DFI水平组(低DFI组:DFI≤30%,439个周期)和高DFI水平组(高DFI组:DFI30%,228个周期);再根据不同受精方式分为两个亚组:常规IVF组及ICSI组,比较不同水平的精子DFI及不同受精方式对FET周期妊娠结局的影响。结果纳入研究的667对不孕夫妇中临床妊娠334例,总妊娠率50.07%。低DFI组、高DFI组患者的一般资料比较均无统计学差异(P0.05),但低DFI组的受精率显著高于高DFI组(P0.05)。低DFI组的临床妊娠率显著高于高DFI组(54.21%vs.42.11%)(P0.05),流产率则显著低于高DFI组(16.81%vs.27.08%)(P0.05)。低DFI组中IVF组和ICSI组的临床妊娠率(52.63%vs.58.62%)、流产率(15.88%vs.19.12%)比较均无显著性差异(P0.05);高DFI组中IVF组的临床妊娠率显著低于ICSI组(33.33%vs.48.48%),流产率则显著高于ICSI组(40.63%vs.20.31%)(P均0.05)。结论精子DFI水平影响不孕患者的受精率;随着精子DFI水平增高,其整体临床妊娠率下降,流产率增加。低DFI水平的不孕夫妇可选择常规IVF或ICSI助孕;但对于高DFI水平的不孕夫妇,选择ICSI助孕方式可能对改善妊娠结局有益。     

不同受精方式对畸形精子症患者体外受精结局的影响

目的依据畸形精子症患者精子浓度来选择采用IVF或ICSI受精方式,探讨其对IVF-ET助孕结局的影响。方法回顾性分析2014年1月至2017年1月于徐州市妇幼保健院生殖医学中心行IVF/ICSI-ET治疗周期的畸形精子症患者,共计82个周期。根据畸形精子症患者精子浓度选择不同的受精方式,分为IVF畸形精子症组和ICSI畸形精子症组,其中精子浓度≥5×106/ml患者行常规IVF治疗(IVF畸形精子症组,44个周期),精子浓度5×106/ml的患者行ICSI治疗(ICSI畸形精子症组,38个周期),对照组为同期行IVF/ICSI-ET助孕治疗精子形态正常的患者,共605个周期(其中IVF对照组494个周期,ICSI对照组111个周期)。分别比较IVF组内、ICSI组内和畸形精子症组内在年龄、不孕年限、子宫内膜厚度及妊娠结局(临床妊娠率、种植率和流产率)等方面的差异。结果 IVF对照组与IVF畸形精子症组,ICSI对照组与ICSI畸形精子症组分别在年龄、不孕年限和移植日子宫内膜厚度均差异不显著(P均0.05);IVF对照组的种植率略高于IVF畸形精子症组,临床妊娠率和流产率均略低于IVF畸形精子症组,但均无统计学差异(P均0.05);ICSI畸形精子症组的临床妊娠率、种植率和流产率高于ICSI对照组(57.9%vs.48.6%;33.8%vs.29.5%;9.1%vs.1.9%),但均无统计学差异(P均0.05);ICSI畸形精子症组的临床妊娠率、种植率高于IVF畸形精子症组(57.9%vs.52.3%;33.8%vs.27.3%),流产率低于IVF畸形精子症组(9.1%vs.13.0%)但均无统计学差异(P均0.05)。结论畸形精子症患者精子浓度≥5×106/ml时,选择常规IVF是可行的。     

不同来源精子对卵胞浆内单精子注射治疗结局的影响

目的:分析不同来源精子对卵胞浆内单精子注射(ICSI)临床结局的影响.方法:接受ICSI治疗的不育夫妇共进行了286个周期,分为3组,A组(射出精子组):射出精液少弱畸(包括严重少弱畸)精子症186个周期;B组(PESA组):经皮附睾抽吸术(PESA)80个周期;C组(TESE组):睾丸精子获取术(TESE)20个周期,比较其妊娠结局.结果:PESA组受精率明显高于射出精子组和TESE组(88%VS 84%,77%,P<0.01),PESA组的临床妊娠率显著高于射出精子组(47%,33%,P<0.05),射出精子组临床妊娠率高于TESE组,但差异无统计学意义;3组的种植率差异极显著(20.7%,31.4%,13.2%,P<0.01);卵裂率(99%,98%,98%)、流产率(7%,15%,0%)均没有显著差异.结论:PESA来源精子比射出精子组及TESE组的受精率及临床妊娠率有所增高,但TESE和PESA来源精子对临床妊娠率没有影响,射出精子组和TESE组来源精子对临床妊娠率没有影响.对于重度少弱精子症患者和梗阻性无精子症患者,进行ICSI治疗,均有机会获得妊娠.     

Zeta potential of human X- and Y-bearing sperm

The zeta potential of human X- and Y-bearing sperm was measured by two different methods: (i) using an electrophoretic light scattering spectrophotometer, and (ii) using a laser-rotating prism. The X- and Y-bearing sperm were separated by free-flow electrophoresis, and their purities were determined by staining for the F-body using quinacrine mustard. The zeta potential of the sperm in the fraction containing more than 80% Y-bearing sperm was approximately -16 mV, whereas that of sperm in the fraction containing more than 95% X-bearing sperm was approximately -20 mV. In other words, the net negative-charge on the cell surface of human X-bearing sperm is higher than that of Y-bearing sperm.     

Role of the epididymis in sperm competition

Although it is generally understood that the testes recruited kidney ducts for reproductive function during the evolution of vertebrates, little is understood of the biological significance of the adaptation. In the context of the evolution of the mammalian epididymis, this report provides evidence that a major role of the epididymis is to enhance a male＇s chance of achieving paternity in a competitive mating system. A unique example of sperm cooperation in monotremes is used as evidence that the epididymis produces sperm competition proteins to form groups of 100 sperm into bundles that have a forward motility nearly thrice that of individual spermatozoa. As it required 3-h incubation in vitro under capacitation conditions to release motile sperm from the bundles, it is suggested that the monotremes provide an example of capacitation that is quite different from capacitation in higher mammals. It is suggested that variation between species in the intensity of sperm competition could explain the variation that occurs between species in the amount of post-testicular sperm maturation and storage in the epididymis, an explanation of why the human epididymis does not play as important a role in reproduction as the epididymis of most mammals. （Asian J Androl 2007 July; 9： 493-499）     

精子尾部卷曲试验及影响因素的探讨

本文介绍了精子尾部卷曲试验及影响因素（时间、温度、ｐＨ）。３０例正常生育男性和４３例不育男性的精液分析表明，精子尾部卷曲率与精子活率具有高度的相关性（ｒ＝０．９６２５），与精子密度无相关性（ｒ＝０．１６２）；男性生育组与不育组之间的精子尾部卷曲率具有显著性差异（ｘ±ｓ，百分率分别为７５．２１±２０．０１和５９．４３±２０．５４，Ｐ＜０．０５）。     

Activation of Spermatozoal Forward Migration in vitro by Hydrocortisone

The effects of hydrocortisone and cortisone on spermatozoal motility in vitro were tested. Hydrocortisone at concentrations of 50, 100 and 1000 nmoles/ml was effective in activating in vitro the forward migration and the motility of boar spermatozoa recovered from the cauda epididymidis. Where boar epididymal spermatozoa were incubated with hydrocortisone at concentrations of 50, 100 and 1000 nmoles/ml for between 0 and 24 h at 25°C in vitro , the spermatozoal motility was significantly higher than where no hydrocortisone was added. With ejaculated boar spermatozoa, hydrocortisone at concentrations of 100 and 1000 nmoles/ml increased the spermatozoal motility for between 0 and 2 h and at a concentration of 50 nmoles/ml increased spermatozoal motility for between 2 and 24 h at 25°C in vitro. After 4 h incubation, the effect of hydrocortisone at a concentration of 100 nmoles/ml on boar ejaculated sperm motility was not significantly different from the control. But, hydrocortisone at a concentration of 1000 nmoles/ml inhibited the forward migration of boar ejaculated sperm after it had been incubated with the sperm for 6 h. Cortisone, although structurally similar to hydrocortisone, had no significant effect in improving the motility of boar spermatozoa.
Both hydrocortisone and cortisone had no demonstrable effect on the forward migration and the motility of human spermatozoa in vitro. ,     

Characterisation of a subpopulation of sperm with massive nuclear damage,as recognised with the sperm chromatin dispersion test

Assessment of human sperm DNA fragmentation by the sperm chromatin dispersion (SCD) test is based on the detection of haloes of spreading DNA loops after sequential DNA denaturing and protamine removal. After the SCD test, sperm without DNA fragmentation show chromatin haloes emerging from the central nuclear core, while sperm containing fragmented DNA present small or no haloes. The nuclear degraded sperm are recognised as a differentiated category within the sperm with fragmented DNA, whose cores appear irregularly and/or faintly stained. This subpopulation is more prevalent in patients with varicocele. Protein staining with 2.7‐dibrom‐4‐hydroxy‐mercury‐fluorescein demonstrated that degraded sperm intensely lose nuclear core proteins after the SCD processing. Moreover, degraded sperm are 65% more faintly labelled for DNA breaks after in situ nick translation (ISNT) on average, due to extensive DNA loss. A two‐dimensional comet assay under sequential neutral and alkaline conditions demonstrated that degraded sperm contain both massive double‐ and single‐strand DNA breaks. The degraded sperm appear as a subpopulation with stronger nuclear damage, affecting both DNA and protein fractions, possibly due to intense intratesticular oxidative stress, what could explain its higher proportion in patients with varicocele.     

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.     

The effect of serum on motility of human spermatozoa in culture

The motility of spermatozoa was higher at 6-22 h in Ham's F10 culture medium supplemented with 20-30% human serum than with lower proportions of serum or with 1% human serum albumin. Heat treatment (56 degrees C, 1 h), charcoal extraction and dialysis (18,000 molecular weight cut-off) of the serum did not reduce sperm motility suggesting that high molecular weight components are responsible for maintenance of motility. Renewing the medium (Ham's F10 with 30% serum) at 7 h resulted in better sperm motility and velocity at 22 h. At 22 h the pO2, pCO2, pH and sodium concentrations were not different in replenished and control cultures, but the concentration of glucose was higher and that of potassium lower if the medium was changed. These results suggest that addition of 20-30% human serum and renewal of medium at intervals is beneficial for sperm culture and may be of use in in vitro fertilization.     

Two Plasminogen Activators, Streptokinase and Urokinase, Stimulate Human Sperm Motility

The stimulatory effects of two plasminogen activators, namely streptokinase and urokinase, were measured with a trans-membrane migration method. Both drugs induced maximal motility increase at a concentration of 200 international unit/ml; the amplitude of maximal motility increase ranged from 17% to 19% of control. Although their stimulatory effects were much less than those of calcium regulating agents, the clinical application of these two drugs for improving the successful rate of artificial insemination deserves further investigation because the action site is seminal plasma rather than sperm.     

Evaluation of different doses of mashua (Tropaeolum tuberosum) on the reduction of sperm production, motility and morphology in adult male rats

Mashua is an edible-tuber crop that grows in the Andean region. Folk medicine describes the use of mashua to reduce reproductive function in men. The present study aimed: (i) to determine whether different doses of mashua (0.01, 0.1, 1 and 2 g kg(-1)) produced a dose-response reduction on sperm production and quality; and, (ii) to determine whether these anti-reproductive effects of mashua can be reversible after cessation of treatment (12 and 24 days of recovery time). Mashua-treated rats showed lower values of daily sperm production, epididymal and vas deferens sperm count and sperm motility; meanwhile, mashua increased the percentage of abnormal sperm morphology and epididymal sperm transit rate. The following variables follow a dose-response effect: sperm number in vas deferens, sperm motility and sperm transit rate. In addition, it was demonstrated that the reduction in reproduction function in male rats treated with mashua was reversible after 24 days of recovery time. Finally, lower doses mashua reduces sperm number and quality (motility and morphology), and these adverse effects on male reproductive system may be reversible after 24 days after cessation of the treatment.     

Sperm selection based on motility in polyvinylpyrrolidone is associated with successful pregnancy and embryo development

The aim of this study was to investigate whether spermatozoon motility in polyvinylpyrrolidone (PVP) is associated with better embryo development and pregnancy rates in ICSI cycles. A total of 123 primary ICSI treatment cycles were included in this study. Semen samples were tested for motility before ICSI procedure in PVP. Within 3 min, the presence or absence of motility was recorded. Sperm functions were examined by the aniline blue (AB) chromatin condensation test and the hypoosmotic swelling test, and the chromatin stability was evaluated by inducing its decondensation with sodium dodecyl sulphate and ethylenediaminetetraacetic acid (EDTA). Fertilisation and embryo scoring were evaluated. Fifty (64%) of 78 women conceived in the PVP (+) group; and 12 (26%) of 45 women conceived in the PVP (?) group; the pregnancy rate was significantly higher in the PVP (+) group (P = 0.003). Semen parameters were observed to be similar in both groups. The mean number of total embryos obtained in ICSI procedure and transferred grade 1 embryos were significantly higher in PVP (+) group (P = 0.01 and P = 0.003 respectively). The presence of sperm motility in PVP is associated with increased pregnancy rate, higher percentage of good quality embryos, sperm chromatin condensation and decondensation.