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。     

附睾及睾丸精子行ICSI治疗无精子症妊娠结局

目的 :回顾性分析 5 0例无精子症患者利用附睾或睾丸精子行卵细胞胞质内单精子注射 (ICSI)的治疗结局。　方法 :经皮附睾精子抽吸术 (PESA)或睾丸切开取精术 (TESE)获得精子行ICSI,评估取精的成功率 ,ICSI后的受精率、种植率及临床妊娠率 ,以精液精子ICSI组作为对照。　结果 :PESA、TESE与精液精子组分别注射MⅡ期成熟卵子 2 86、36 0、15 6 9个 ,受精率 3组差异无显著性 (74 .8% ,75 .2 %vs 77.5 % ,P >0 .0 5 )。种植率、妊娠率TESE与精液精子组差异无显著性 (2 9.87%vs 2 9.5 4 % ;4 8.15 %vs 5 2 .6 0 % ,P >0 .0 5 ) ,PESA组显著高于TESE组及精液精子组 (5 0 .85 %vs 2 9.87% ,2 9.5 4 % ;6 8%vs 4 8.15 % ,5 2 .6 0 % ,P <0 .0 5 )。PESA组共妊娠 17例 ,已分娩 6例 ,继续妊娠 9例 ,流产 2例 ;TESE组共妊娠 13例 ,已分娩 7例 ,继续妊娠 4例 ,流产 2例。　结论 :采用附睾或睾丸精子行ICSI是治疗男性无精子症的有效方法。     

无精子症病人100例取精方法及妊娠结局

目的 :回顾性分析 2 0 0 1年 1月～ 2 0 0 2年 1月在生殖中心行卵胞质内单精子注射 (ICSI)治疗的 10 0例无精子症男性的治疗结果。　方法 :经皮附睾精子抽吸术 (PESA)或睾丸精子抽提术 (TESE)获得精子 ,女方进行常规超排卵。分析激素水平 ,行睾丸组织学检查 ,评估取精的成功率、受精率、种植率和临床妊娠率。　结果 :76例(76 % )经PESA获得精子 ,2 3例 (2 3% )通过TESE获得精子。PESA和TESE组的受精率、种植率和临床妊娠率分别为 71.3%和 75 .18% ,2 0 .35 %和 2 2 .0 5 % ,4 2 .11%和 4 1.6 0 %。PESA组有 32例临床妊娠 ,其中 15例继续妊娠 ,15例已分娩 ,2例流产。TESE组有 10例临床妊娠 ,其中 6例继续妊娠 ,2例已分娩 ,2例流产。两组的受精率、种植率和临床妊娠率差异无显著性。在TESE组有 1例取精失败而放弃治疗。　结论 :激素水平和睾丸组织学检查不能预测附睾或睾丸取精的成功 ,PESA和TESE获得精子进行单精子注射是治疗男性无精子症的有效方法 ,两组的受精率 ,种植率和临床妊娠率差异无显著性 (P >0 .0 5 )。     

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。     

冻融复苏微量精子行卵细胞胞质内单精子注射术的疗效及临床妊娠结局分析

目的:回顾性分析123例无精子症患者经皮附睾精子抽吸术(PESA)或经皮睾丸精子抽吸术(TESA)后冻融复苏微量精子行卵细胞胞质内单精子注射术(ICSI)的疗效及临床妊娠结局情况。方法:将采用微量冻融PESA、TESA精子行ICSI的病例归为冻融精子组,采用新鲜PESA、TESA精子行ICSI的病例归为对照组。比较冻融精子组与新鲜精子组组间及组内的双原核(2PN)受精率、优质胚胎率、临床妊娠率、流产率、宫外孕率、多胎妊娠率有无统计学差异。结果:PESA精子冻融组与新鲜组受精率、优质胚胎率、临床妊娠率、流产率、宫外孕率及多胎妊娠率分别为75.67%vs76.49%,64.96%vs66.19%,55.21%vs57.22%,13.21%vs12.61%,3.77%vs5.41%,37.74%vs37.84%(P>0.05),TESA精子冻融组与新鲜组受精率、优质胚胎率、临床妊娠率、流产率、宫外孕率及多胎妊娠率分别为74.41%vs76.43%,64.63%vs66.35%,46.81%vs53.39%,18.18%vs14.55%,4.55%vs1.82%,37.74%vs37.84%,组间及组内均无统计学差异(P>0.05)。PESA精子与TESA精子冻融复苏成功率为70.07%vs62.67%,无统计学差异(P>0.05)。结论:微量PESA及TESA精子冻融技术对无精子症患者来说是一种安全、经济、有效的治疗方法;精子冷冻复苏技术有待于进一步提高;该技术是否会增加子代远期遗传风险仍有待于进一步探讨和研究。     

不同方式获取精子对ICSI治疗后妊娠结局分析

目的 分析和比较3种不同获取精子方式行ICSI后的妊娠结局.方法 回顾性分析本中心2009年2月至12月进行ICSI治疗的207 例患者,其中严重少弱精子患者108例、当天受精失败81例和经皮附睾穿刺术(PESA) 18例,比较3组的受精率、卵裂率、优质胚胎率和妊娠率.结果 3组的受精率、卵裂率、优质胚胎率无明显差异,其中受精失败组行补救ICSI妊娠率较高,但与其他两组比较没有统计学意义.结论严重少弱精子症、附睾穿刺和补救ICSI获得的精子,受精率和优质胚胎率,妊娠率相似,不同来源和生理功能的精子不影响ICSI的治疗效果.     

146例炎症梗阻性无精子症的临床评估和ICSI治疗结局分析

目的分析炎症梗阻性无精子症的临床评估和单精子卵胞浆内注射(ICSI)的治疗结局。方法前瞻性研究近5年间接受ICSI治疗的炎症性梗阻性无精子症的临床特征、精液和超声特点,经皮附睾穿刺精子抽吸术(PESA)或经皮睾丸穿刺取精术(TEFNA)结合ICSI治疗后观察受精、临床妊娠等结果。结果146例患者体检附睾均有增粗变硬或伴头尾部结节。82例患者曾有生育史、附睾炎症史或输精管附睾吻合手术史,其中72例PESA找到附睾精子;53例无上述病史者49例PESA找到附睾精子:另有精道远端梗阻11例。ICSI治疗146例167周期炎症性梗阻性无精子症的受精率、每周期临床妊娠率分别为81.1%和42.1%。结论炎症梗阻性无精子症具备典型的临床和超声特征,PESA附睾精子获取率高,ICSI治疗获得较高受精率和临床妊娠率。     

中、重度少弱精子症患者冻融精子与新鲜射出精子的ICSI临床结局比较

目的研究中、重度少弱精子症患者冻融精子与新鲜射出精子对ICSI结局的影响。方法回顾性分析227对不育夫妇240个ICSI治疗周期,分为3组:新鲜重度少弱精子组(A组,107例,111个周期);新鲜中度少弱精子组(B组,71例,76个周期);冻融中、重度少弱精子组(C组,49例,53个周期)。比较各组的临床结果。结果新鲜重度、中度少弱精子组的可用胚胎率(分别为78.1%、75.0%)、优质胚胎率(分别为46.1%、46.2%)显著高于冻融组(分别为66.7%和37.4%)(P0.05),但3组间受精率、正常受精率、卵裂率、临床妊娠率、种植率均无显著性差异(P0.05)。新鲜中度少弱精子组与新鲜重度少弱精子组在可用胚胎率、优质胚胎率、受精率、正常受精率、卵裂率、临床妊娠率及种植率方面均无统计学差异(P0.05)。结论中、重度少弱精子症患者冻融精子行ICSI影响可用胚胎率和优质胚胎率,但不影响临床妊娠率和胚胎种植率。     

经皮附睾精子抽吸术和睾丸精子获取术在无精子症诊断和治疗中的应用

目的 :研究附睾和睾丸精子抽吸术对无精子症患者的诊断和治疗价值。　方法 :应用经皮附睾精子抽吸术(PESA)和睾丸精子获取术 (TESE)两种方法对 385例无精子症患者进行穿刺检查。　结果 :其中 6 4例附睾中存在精子 (1 6 .6 2 %) ;4 5例患者睾丸中存在精子 (1 1 .6 9%) ;对其中 6 4例睾丸或附睾中发现精子的患者采取PESA或TESE取精后行卵细胞胞质内单精子注射 (ICSI)治疗。胚胎移植后妊娠率为 39.0 7%。　结论 :PESA和TESE为部分无精子症患者提供了生育的机会 ,也是针对无精子症的有效的治疗手段。     

不同来源精子行卵胞浆内单精子注射临床妊娠结局分析

目的探讨不同来源精子对卵胞浆内单精子注射(ICSI)治疗后妊娠结局的影响。方法回顾性分析因男性因素行ICSI的不育患者835例的859个助孕周期,根据精子来源不同分A组即射精精子组699周期,B组即经皮附睾精子抽吸术(PESA)组114周期,C组即睾丸精子抽吸术(TESA)组46周期。结果 (1)A、B、C三组正常受精(2PN)率分别为:82.4%、76.2%、71.8%,A组最高,即射精组PESA组TESA组,B、C组与A组比较有显著性差异(P0.05);A、B、C三组卵裂率分别为:98.6%、97.6%、98.3%,B组与A组比较有显著性差异(P0.05),而C组与A组比较无显著性差异(P0.05)。(2)A、B、C三组可利用胚胎率、优质胚胎率比较无显著性差异(P0.05)。(3)A、B、C三组着床率分别为32.5%、37.7%、41.7%,三组比较无显著性差异(P0.05)。(4)A、B、C三组妊娠率分别为50.8%、56.1%、60.9%,三组比较无显著性差异(P0.05)。结论不同来源精子行ICSI,对受精率有影响,但三组都可获得相似的可利用胚胎、优质胚胎,应用射精精子以及PESA、TESA精子行ICSI都能获得较好的妊娠结局。     

Computerized sperm motility and application of sperm cryopreservation

An objective method for measuring sperm motion characteristics was developed on an Intellect 100 Quantel Image Analysis System suitable for various image cytometric applications. It provided overall analysis of percent motility (% MS) as well as individual and mean measurements of motion characteristics, including vigor characteristics such as curvilinear velocity (Vc), straight line velocity (Vsl), and trajectory pattern characteristics, that is, progressiveness ratio (PR) and amplitude of lateral head displacement (Alh). Evaluation of the method for reproducibility and accuracy showed reliable measurements of these parameters measured on a minimum of 70 motile sperm, sufficient to describe adequately the sperm population. A study was performed comparing motion characteristics of 30 semen samples falling in a normal range before and after cryopreservation in cryoprotector medium (CM). A mean motility rate of recovery (MRR) of 45% was obtained. Only sperm count and concentration in motile forms among initial semen variables correlated weakly with MRR. Velocity recovery rate (VRR) approached 1 with a marked variability among ejaculates. Distribution profile of Vc was highly modified by freezing in CM: spermatozoa that were initially fast and progressive were the most resistant to cryoaggression. PR and Alh values were little affected by freezing in CM. The tolerance of various samples from a given patient was highly variable for % MS and Alh and less variable for Vc and PR. This illustrates the difficulty in predicting the effect of freezing on motility characteristics and, therefore, of extrapolating from semen variables the ability of frozen-thawed samples to fertilize.     

Accurate sperm morphology assessment predicts sperm function

Sperm morphology has been associated with in vitro as well as in vivo fertilisation. The study aimed to evaluate the possible relation between the percentage of spermatozoa with normal morphology and the following sperm functional assays: (i) zona-induced acrosome reaction (ZIAR); (ii) DNA integrity; (iii) chromatin condensation; (iv) sperm apoptosis; and (v) fertilisation rates. Regression analysis was employed to calculate the association between morphology and different functional tests. Normal sperm morphology correlated significantly with the percentages of live acrosome-reacted spermatozoa in the ZIAR (r = 0.518; P < 0.0001; n = 92), DNA integrity (r = -0.515; P = 0.0018; n = 34), CMA(3) -positive spermatozoa (r = -0.745; P < 0.0001; n = 92), sperm apoptosis (r = -0.395; P = 0.0206; n = 34) and necrosis (r = -0.545; P = 0.0009; n = 34). Negative correlations existed between for the acrosome reaction, and DNA integrity, while negative associations were recorded with the percentages of CMA(3) -positive spermatozoa, apoptotic and necrotic spermatozoa. Sperm morphology is related to sperm dysfunction such as poor chromatin condensation, acrosome reaction and DNA integrity. Negative and significant correlations existed between normal sperm morphology and chromatin condensation, the percentage of spermatozoa with abnormal DNA and spermatozoa with apoptotic activity. The authors do not regard sperm morphology as the only test for the diagnosis of male fertility, but sperm morphology can serve as a valuable indicator of underlying dysfunction.     

Efficacy of sperm wash media in improving sperm motility

Spermatozoa from 15 fertile men were washed with Ham's F10 and incubated with two commercially available sperm nutrient media for 2, 4, 6, and 24 h. Both sperm capacitation medium (Irvine Scientific Co., Santa Ana, CA) and Pro-ception (Milex Products, Inc., Chicago, IL) proved to be capable of improving sperm motion characteristics. These media may be used for incubating sperm for intrauterine insemination or for in vitro fertilization.     

附睾、睾丸精子卵浆内单精子注射治疗阻塞性无精症引起不育

为评价附睾或睾丸精子卵浆内单精子注射（ＩＣＳＩ）治疗阻塞性无精症引起不育的疗效，对３１例无精子男性不育患者配偶进行超排卵治疗３７个周期，获卵当日从患者附睾取精，其中２４例获得活动精子，７例失败，改用钳取睾丸曲细精管从中分离精子以供ＩＣＳＩ。结果：共获卵４５３个，附睾、睾丸精子ＩＣＳＩ受精率分别为５５７％、６１７％，平均每周期移植胚胎３７个，总妊娠率每周期２９７％。结论：只要获得活动精子ＩＣＳＩ，阻塞性无精症患者也有机会生育     

Effects of altered epididymal sperm transit time on sperm quality

The epididymal sperm transit time seems to have an important role in the process of sperm maturation, and it seems that alterations to the transit can harm the process. The aim of the present work was to evaluate the influence of altered sperm transit time through the epididymis on sperm parameters and fertility of rats, as well as the role of testosterone in the alterations. Sprague–Dawley adult male rats were randomly assigned to four different groups and were treated for 12 days: (i) 10 μg/rat/day DES, to accelerate the transit; (ii) 6.25 mg/kg/day guanethidine sulphate, to delay the transit; (iii) same treatment as group 1, plus androgen supplementation; (iv) control animals received the vehicles. Guanethidine treatment delayed the sperm transit time through the epididymal cauda, provoking increased sperm reserves in this region. Animals exposed to DES showed an acceleration of sperm transit time in the epididymis, and consequently decreased sperm density in both epididymal regions, the caput-corpus and cauda, and diminished sperm motility. In both cases sperm production was not altered. Testosterone supplementation was able to restore the transit time to values close to normality, as they were higher than in the control rats. The same occurred in relation to sperm motility. Rats exposed to DES presented lower fertility after in utero artificial insemination using sperm collected from the proximal cauda epididymis. Therefore, it was concluded that the acceleration of rat sperm transit time appeared to harm normal sperm maturation, thus decreasing sperm quality and fertility capacity, in an androgen-dependent way.     

Efficiency of percutaneous testicular sperm aspiration as a mode of sperm collection for intracytoplasmic sperm injection in nonobstructive azoospermia

PURPOSE: We determined the feasibility of obtaining mature spermatozoa for intracytoplasmic sperm injection (ICSI) by percutaneous testicular sperm aspiration in men with nonobstructive azoospermia. We also compared the results of ICSI using spermatozoa recovered by open excisional biopsy versus percutaneous testicular sperm aspiration. MATERIALS AND METHODS: A total of 84 men with nonobstructive azoospermia underwent percutaneous testicular sperm aspiration to recover testicular spermatozoa for ICSI on the day of ova retrieval from the wife. Percutaneous testicular sperm aspiration was performed with the patient under general anesthesia in the upper and lower poles of each testis. It was followed by immediate microscopic search of the aspirate to confirm the presence of spermatozoa. In the absence of spermatozoa open excisional biopsy was performed in the same setting. RESULTS: Percutaneous testicular sperm aspiration resulted in the recovery of mature spermatozoa in 45 men (53.6%). Of the remaining 39 men (46.4%) requiring open biopsy adequate spermatozoa were recovered in 28 (71.8%). Although the fertilization rate was significantly higher in the sperm aspiration group, the cleavage and pregnancy rates were similar in the 2 groups. CONCLUSIONS: Percutaneous testicular sperm aspiration was a successful initial approach to collect mature spermatozoa in a high proportion of men with nonobstructive azoospermia. It is safe, minimally invasive and well tolerated by all patients.     

Temperature-induced sperm nuclear vacuolisation is dependent on sperm preparation

The influence of temperature during incubation on the degree of sperm nuclear vacuolisation was assessed by two different experiments. In a first experiment, motile spermatozoa from 24 patients were prepared by the swim-up technique and incubated either at room temperature or at 37 °C for up to 4 h. The presence of sperm nuclear vacuoles was determined by contrast-enhanced high magnification microscopy. No statistically significant difference was found in the degree of sperm nuclear vacuoles in both groups (RT: 45.6 ± 17.6%; 37 °C: 48.4 ± 17.0%) following 4 h of incubation. In a second experiment, spermatozoa from six patients were either prepared by swim-up or washed and incubated at 37 °C. After 4 h of incubation, a significant increase in sperm nuclear vacuolisation was found in washed sperm (from 51.5 ± 15.4% to 68.6 ± 9.0%; P < 0.05) but not in swim-up sperm (from 51.5 ± 15.4% to 48.2 ± 17.1%; n.s.). Our data show that the mode of sperm preparation does influence sperm nuclear vacuolisation at 37 °C (Experiment II). However, sperm nuclear vacuolisation is unaffected by temperature in motile sperm after preparation and isolation by swim-up.     

The effect of sperm DNA fragmentation on intracytoplasmic sperm injection outcome

Our study objective was to assess the effect of various sperm DNA fragmentation levels on clinical intracytoplasmic sperm injection outcome. This retrospective study included 392 patients who underwent ICSI and performed sperm DNA fragmentation testing before the procedure. Based on sperm DNA fragmentation cut-off values, the patients were differentiated into 3 groups as <20%, 20%–30% and >30%. According to the female status, patients were differentiated into favourable group (n = 259) with female age <35 years and anti-Mullerian hormone level ≥7.1 pmol/L; and unfavourable group (n = 133) with female age ≥35 years and anti-Mullerian hormone level ≤7.1 pmol/L. The patient's medical records were reviewed, and patient's demographic, laboratory data including semen analysis, sperm DNA fragmentation determined by means of sperm chromatin dispersion, hormonal profile and data regarding intracytoplasmic sperm injection cycle were collected. This cohort reported that the clinical reproductive outcomes of intracytoplasmic sperm injection showed no statistical significance with increase sperm DNA fragmentation levels. In sperm DNA fragmentation above 30%, favourable females had significantly higher clinical pregnancy rate and live birth rate than unfavourable females, while fertilisation rate and miscarriage rate showed no significance between the subgroups. High sperm DNA fragmentation is linked to poor semen parameters.     

Stability of sperm characteristics in men with disturbances in sperm quality

Sixty-one men referred to our laboratory for semen analysis, and subsequently judged to exhibit some form of sperm pathology, were asked to return for a second analysis, not less than 2 months after the first, in order to assess the stability of the pathological changes observed. In almost half of the cases, the referring physician had, on his own initiative, started hormone or antibiotic treatment. The sperm parameters studied included sperm count, sperm motility judged by laser-Doppler spectroscopy, and sperm morphology and viability. The motility characteristics included percentage motile, their average velocity, and percentage swimming in a progressive manner, and their progressive velocity. In untreated subjects, there was no significant difference between the first and second analysis in any of the sperm parameters measured. This was also true for both oligozoospermic individuals (less than 20 x 10(6) sperm/ml) and the group with higher sperm concentrations. All parameters were highly correlated on the two occasions. The average coefficients of variation of the paired observations were highest for sperm count (approximately 25%) and lowest for sperm velocities and the proportion of abnormal and viable cells in the ejaculate (1-9%). No major differences in the extent of variation could be detected between the low and high sperm density groups. In general, the unsystematic antibiotic and hormone regimens (clomiphene or androgen) used by the referring physicians had no discernable effect on any aspect of sperm quality, indicating the need for more controlled and standardized programmes of treatment.     

精子计数池深度对精子活力影响的实验研究

目的:调查精子计数池深度对精子活力检测结果的影响。方法:利用Filmetrics间隙测量仪精确测量4种不同深度的Geoffrey精子计数池,然后按照WHO手册第5版的要求利用瑞祺CFT-9201型精子质量检测分析系统分析36例精液标本的前向运动精子百分率(PR)、非前向运动精子百分率(NP)和总活动率(PR+NP),并进行比较分析。结果:4种精子计数池的深度分别为9.8、12.7、15.7和19.9μm,测得的PR分别为(44.00±11.63)%、(41.96±12.62)%、(40.86±11.71)%和(37.78±11.38)%,NP分别为(13.54±3.01)%、(14.13±2.94)%、(14.91±3.02)%和(16.53±2.77)%,总活动率分别为(57.53±11.06)%、(56.08±11.97)%、(55.78±11.55)%和(54.31±12.11)%。精子计数板深度与PR呈显著负相关(r=-0.993,P<0.05),与NP呈显著正相关(r=0.989,P<0.05),与活动率呈显著负相关(r=-0.978,P<0.05)。尽管精子总活动率在4种不同深度的计数池之间没有显著差异,但PR和NP在9.8μm深的计数池和19.9μm深的计数池之间均有显著性差异(P<0.05)。结论:精子计数池的深度对精子活力的影响不容忽视,不同深度计数池获得的结果差异将会给临床医生对患者做出正确的诊断(如弱精子症)和采取合适的治疗措施带来一定的负面影响。不同深度的精子计数池应有相应的精子活力正常参考值范围。