圆头精子的冷冻保存

目的:探讨圆头精子的冷冻保存效果及其精子功能改变。方法:6例圆头精子症患者各提供1-3份精液标本,应用不含卵黄冷冻保护剂和二步降温方法冷冻保存圆头精子症标本,检测冷冻精子活动率、头-尾膜完整率、存活时间和顶体蛋白酶活性。结果:圆头精子标本冷冻保存后,精子活动率显著减少,膜完整率显著下降,头膜损伤-尾膜完整的精子率显著升高(n=13,P＜0.01)。体外存活时间缩短。精子冷冻前后无顶体蛋白酶活性。不含卵黄冷冻保护剂与含卵黄冷冻保护剂,以及两步冷冻与多步冷冻的效果比较未见显著差异(n=7,P＞0.05)。结论:圆头精子可经历冷冻保存后存活,但冷冻复苏率低。冷冻-解冻过程显著降低精子功能。不含卵黄冷冻保护剂和两步降温可应用于冷冻保存圆头精子症标本。     

冷冻保存对正常和不育症精子PH-20表达和凋亡的影响

目的探讨冷冻保存对人精子膜蛋白PH-20表达和精子凋亡的影响。方法 14例正常生育力精液标本(A组)和20例不育症精液标本(B组)行冷冻保存。Western blotting检测PH-20蛋白在人精子中的表达,免疫荧光用来观察PH-20蛋白在人精子上的定位,应用末端脱氧核苷酸转移酶(Td T)介导的原位末端标记(TUNEL)法检测精子凋亡情况。结果解冻后正常生育组和不育组的PH-20/β-actin平均吸光度与冷冻前比较均有显著性下降(P0.05);解冻后正常生育组和不育组的PH-20阳性率与冷冻前比较均有显著性下降(P0.05)。解冻后正常生育组的精子凋亡率与冷冻前的比较差异无显著性意义(P0.05)。而解冻后不育组的精子凋亡率与冷冻前的比较均显著性下降(P0.05),且不育组的降低程度大于正常生育组。结论冷冻-解冻过程可引起精子PH-20蛋白表达减少和精子PH-20阳性率降低,但冷冻保存对正常生育者的精子凋亡率无显著影响。     

冷冻保存对人类精子顶体完整性及超微结构的影响

目的探讨冷冻保存对人类精子顶体完整性及超微结构的影响.方法 2 0例正常生育力精液标本(A组)和27例不育症精液标本(B组)行冷冻保存.应用荧光标记豌豆凝集素法检测冷冻前、后精子顶体完整率.采用透射电镜观察冷冻精子头部超微结构的改变 (n=3).结果解冻后A和B组的顶体完整率与冷冻前的比较均有显著性下降(P<0.01) ,且B组的降低程度明显大于A组.透射电镜观察到冷冻精子头部超微结构发生不同程度的损伤,浆膜和顶体膜出现肿胀、破损,顶体结构异常改变显著增多,顶体内容物丢失,甚至顶体帽缺失.结论冷冻-解冻过程对精子顶体造成了损伤,引起顶体完整率降低和超微结构改变.     

冷冻保存对人精子膜蛋白PH-20和透明质酸酶活性的影响

目的:探讨冷冻保存对人精子膜蛋白PH-20和顶体内透明质酸酶活性的影响。方法:24例正常生育力精液标本行冷冻保存。免疫印迹检测PH-20蛋白在人精子中的表达,免疫荧光观察PH-20蛋白在人精子上的定位,采用改良透明质酸钠-明胶底物膜法检测精子顶体内透明质酸酶(HYD)活性(HYD阳性反应率、HYD活性强度)。结果:免疫印迹显示正常标本冷冻前、后精子均有PH-20蛋白的表达,其PH-20/β-actin平均光密度在正常组冷冻前为0.41±0.15;正常组解冻后为0.24±0.11,两者的平均光密度差异有统计学意义;间接免疫荧光染色后在荧光显微镜下观察冷冻前PH-20阳性率达72.88%±8.04%;解冻后PH-20阳性率降至46.97%±8.38%,差异有统计学意义;解冻后HYD活性与冷冻前比较均有显著性下降。结论:冷冻-解冻过程可引起精子PH-20蛋白表达减少和精子PH-20阳性率降低,以及受精过程中的关键酶-HYD活性减弱。     

精子形态分析在男性不育诊断中的应用价值

目的探讨精子形态染色技术在男性不育诊断中的应用价值。方法已生育健康男性精液标本20例(对照组)、不孕组男性患者精液86例(A组),不良孕产史组男性患者精液标本65例(B组),精液经处理后涂片,用WHO推荐的D iff-Qu ik快速染色方法对精子进行形态染色,计数正常及异常形态精子,并计算精子正常形态的百分率。结果对照组精液标本中精子正常形态率为49.63%,不孕组精子正常形态率为19.34%,显著低于对照组,两组相比有极显著性差异(P<0.01)。不良孕产史组精液标本中精子正常形态率为42.84%。与对照组相比有显著性差异(P<0.05),与不孕组相比有极显著性差异(P<0.01)。结论精子形态染色在临床男性不育的诊断中占有重要地位。在进行常规精液分析时,增加形态学染色项目,对男性不育患者的诊治有很大帮助,有助于临床为不孕患者选择助孕方式,为有不良孕产史夫妇查找病因,作为临床诊治的依据。     

人精液一氧化氮含量与男性不育的相关性

目的:研究精液中一氧化氮(NO)含量对精子凋亡的影响及其与男性不育之间的相关性.方法:采用精子质量检测系统对精液标本进行常规检查;应用硝酸还原酶法测定精液中NO的含量;应用瑞-姬染色和TdT介导的原位末端标记(TUNEL)法检测凋亡精子;观察凋亡精子的形态结构改变.结果:不育组精液中NO(58 37±14 14)μmol/l比正常生育组精液中NO(35 20±8 23)μmol/l含量高;正常生育组精子凋亡率9 67%±2 54%比不育组精子凋亡率33 98%±10 54%低.结论:不育组精液中精子的凋亡率随着NO含量的增高而增加.高浓度的NO导致精子凋亡率增加而致使男性生育力下降.     

弓形虫感染对男性精液质量的影响

目的探讨弓形虫感染对男性精液质量的影响。方法应用伟力彩色精子质量分析系统对86例弓形虫感染的不育男性和64例正常生育男性精液的精液量、液化时间、精子总数、精子密度、精子活力、精子活率等参数进行分析。结果正常男性组与弓形虫感染男性组精液的精液量、液化时间、精子总数两组无明显差异（P〉0.05）;精子密度、a级精子百分率、a＋b级精子百分率、精子活率两组差异有显著性（P〈0.05-0.01）。结论弓形虫感染可引起男性精液质量异常,为阐明弓形虫感染引起男性不育的机制提供了科学依据。     

精子形态学分析在男性不育症诊断中的应用

目的探讨进行精子形态学分析在诊断男性不育症中的应用。方法严格按照WHO技术规范,对来我院男科门诊就诊的3553例男性患者精液进行常规分析,同时进行精子形态学分析。结果若以精子密度≥20×106/ml,精子活动率:a级≥25%或a+b级≥50%为标准,精液质量正常者1861例,但其中却有462例精液正常形态精子百分率<15%。结论精子形态异常是影响男性生育力的重要因素,将精液常规与精子形态学分析相结合,有助于提高男性不育症的诊断率。     

精液白细胞与精子形态的相关性研究

目的探讨精液白细胞与精子形态之间的关系。方法按WHO精液检测方法对192例不育患者进行精子形态分析和精液白细胞检测。结果白细胞精子症组正常精子形态百分率低于非白细胞精子症组（P〈0．05）,且白细胞精子症组头部缺陷数和尾部缺陷数较非白细胞精子症组增加,差异有统计学意义。结论精液中过多的白细胞可导致精子形态异常,是男性不育重要的原因之一。     

精子形态与顶体酶活性关系的研究

目的探讨精子形态与精子顶体酶活性的关系。方法对395例男性不育患者采用精子形态检测系统下人工修正方法分析精子形态,用酶标法检测精子中的顶体酶和顶体酶活力指数（AAI）。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,两组间较差异极显著（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=0.169,P〈0.001）。结论精子形态可影响精子顶体酶活性。     

Predictive value of classical and automated sperm analysis for in-vitro fertilization.

The fertilization rates observed in 122 attempts at in-vitro fertilization were examined in relation to sperm characteristics assessed by visual and automated screening. Using linear regression analysis, a significant correlation was found between the fertilization rate and (i) evaluations in fresh semen sperm concentration, percentages of sperm motility, vitality and normal morphology and velocity, (ii) measurements in swim-up preparations of percentages of sperm motility, vitality and morphology, velocity and amplitude of lateral head displacement. No significant correlation was found between the fertilization rate and any of the parameters studied in 24-h-old swim-up suspensions. Analysis by multiple variable stepwise linear regression showed an optimal correlation (R6 = 0.62) between the observed fertilization rate and theoretical calculation obtained from the following predictive function: fertilization rate = -0.3 + (0.008 x swim-up motility) + (0.004 x normal sperm morphology in fresh semen). Introduction of kinematic characteristics studied by automated screening improved the multiple correlation between the calculated and observed fertilization rate in cases of normal or mildly defective semen. Because of the limited availability of motile spermatozoa, automated analysis could not supersede classical sperm analysis in cases of more severe sperm defects.     

精子DNA碎片与活性氧的关系研究及对IVF结局的影响

目的探讨不育男性的精子DNA损伤程度与精液常规分析各项指标及活性氧的关系,以及对常规体外受精-胚胎移植(IVF-ET)结局的影响。方法接受常规IVF受精的夫妇135例,男性患者作为研究对象,按照精子DNA损伤率将患者分为3组:正常组(精子DNA损伤率0%～15%)79例、轻度损伤组(精子DNA损伤率15%～30%)49例、重度损伤组(DNA损伤率>30%)7例。分析精子DNA损伤对精液常规指标(体积、浓度、活动力和形态)的影响,观察三组患者精浆中过氧化氢(H2O2)、过氧化氢酶(CTA)浓度的变化及三组间受精率、卵裂率、优胚率、种植率、临床妊娠率的差异性。结果重度损伤组的精子活力(a+b)明显低于正常组和轻度损伤组(P<0.05),重度损伤组及轻度损伤组的精子畸形率明显高于正常组(P<0.05)。重度损伤组的优胚率明显低于正常组和轻度损伤组(P<0.05),三组间受精率、卵裂率、植入率和临床妊娠率无统计学差异。重度损伤组精浆中的H2O2浓度明显高于正常组和轻度损伤组(P<0.05),而CTA浓度无统计学差异。结论精子DNA损伤与高H2O2水平有关。精子DNA损伤可导致精液的活动力下降,精子畸形率增高,降低优胚率,不利于IVF-ET的结局。     

中山地区应用体外受精技术治疗不育症的初步报告

目的：初步报告中山市应用体外受精技术（IVF）治疗不育症的效果。方法：28对不育夫妇的男方在精液常规检查的基础上,进行精子形态学和顶体状态分析。28例受术妇女采用Buserelin/FSH/HCG黄体期长方案控制超排卵,并根据不孕病因、患者年龄及卵巢基础FSH／LH比值指导控制性超排卵用药,比较周期取卵率、受精率、卵裂率及临床妊娠率。结果：基础FSH／LH比值正常反应组问收卵率高于低反应组（P＜0．05）。取卵291个,平均每周期取卵9．6个;受精率81．8％;卵裂率79．8％。9例临床妊娠,5例生化妊娠,妊娠率32％。结论：IVF技术是治疗不育症的有效手段,本文只做丈夫精子IVF,其正常精子形态率和顶体完整率对预测受精有指导意义,根据基础FSH／LH比值判断卵巢储备功能及指导超排卵用药,可避免过大、过少剂量对患者的不利影响。     

Optimal management of extreme oligozoospermia by an appropriate cryopreservation programme

BACKGROUND: Severe oligozoospermia is characterized by sperm count fluctuations that may result in insufficient quantities of motile sperm for ICSI on the day of oocyte retrieval, thus necessitating testicular biopsy. To avoid this, we proposed that patients, with transient azoospermia or repeatedly low sperm counts, make a safety pool of frozen spermatozoa before ICSI attempts. METHODS: Seventy cryptozoospermic (<10(3) spermatozoa/ml) and 46 oligozoospermic patients (10(3)-10(5)/ml) were included. Although all oligozoospermic patients succeeded in sperm banking, only 44 of 70 cryptozoospermic patients were successful. Others underwent testicular extraction of spermatozoa. The ICSI results for frozen sperm from cryptozoospermic patients were compared with those obtained with fresh sperm from a group of normal patients (>10(5) spermatozoa/ml). RESULTS: In this prospective matched, controlled study, five cryptozoospermic, but no oligozoospermic, patients failed to produce sperm on the ICSI day, and frozen sperm was used instead. Although fertilization and pregnancy rates (per attempt) using fresh (49% and 5/44, respectively) and frozen sperm (54% and one-fifth, respectively) were similar for this cryptozoospermic group, the results for fresh sperm were significantly lower when compared with the control group (66% and 16/43, P < 0.0001, P < 0.001, respectively). In contrast, results for the oligospermic and control groups were similar. CONCLUSIONS: Banking of ejaculated sperm is helpful for cryptozoospermic patients.     

Predictive value of normal sperm morphology in intrauterine insemination (IUI): a structured literature review

The aim of the study was to conduct a structured review of the literature published on the use of normal sperm morphology, as an indicator of male fertility potential in intrauterine insemination (IUI) programmes. Published literature in which normal sperm morphology was used to predict pregnancy outcome in IUI during the period 1984-1998 was reviewed. In total, 421 articles were identified via Medline searches. Eighteen provided data that could be tabulated and analysed. Eight of the analysed studies provided sufficient data for statistical analysis, six studies used the Tygerberg 'strict' criteria, and two the WHO guidelines (1987, 1992). A meta-analysis of the six studies in the strict morphology group yielded a risk difference (RD) between the pregnancy rates achieved in the patients below and above the 4% strict criteria threshold of -0.07 (95% CI: -0.11 to 4.03; P<0.001). The WHO criteria group (1987, 1992) had insufficient data to be analysed. Meta-analysis showed a significant improvement in pregnancy rate above 4% threshold for strict criteria. Accurate evaluation of normal sperm morphology results should be an integral part of evaluating the male factor.     

Relationship between human sperm-hyaluronan binding assay and fertilization rate in conventional in vitro fertilization

BACKGROUND: Sperm-hyaluronan-binding assay (HBA) is one of the commercial kits being marketed for routine testing of sperm maturity and fertility. However, there is no report of whether the HBA can provide additional information over standard semen analysis for sperm-fertilizing ability. The objective of this study was to investigate the relationship between HBA and fertilization rate in conventional IVF. METHODS: A total of 175 IVF patients with > or = 3 mature oocytes inseminated were included in the study. Both the standard semen analysis and the HBA were performed on the same ejaculated sperm samples used for IVF treatments. Relationships between the semen analysis and the HBA results and fertilization rate were analysed by both the Spearman test and the multivariate logistic regression analysis. RESULTS: Both total and progressive sperm motility and normal morphology were highly correlated with HBA scores. While both normal sperm morphology and HBA scores were statistically significantly related to fertilization rates, the HBA was less significant than normal sperm morphology. The HBA does not provide additional information for identifying patients with a poor fertilization rate. CONCLUSION: HBA is highly significantly correlated with sperm motility and morphology but is less significant than sperm morphology in relation to the fertilization rate in IVF. Thus, the clinical predictive value of HBA for sperm-fertilizing ability in vitro is limited.     

What constitutes a normal seminal analysis? Semen parameters of 243 fertile men

A cross-sectional study was conducted to determine the semen parameters (i.e. volume, concentration, motility, viability and normal morphology) of proven fertile males in Singapore and compare it with the World Health Organization (WHO) recommended normal values and to examine some factors that may affect spermatogenesis. A total of 243 men, whose wives were pregnant at the time of collection of semen, provided a semen sample each after sexual abstinence for 3 days. A questionnaire was used to elicit occupational exposure, alcoholic consumption, smoking history and past significant medical history. Most subjects had normal sperm volume (56.4%), concentration (79.8%), motility (69.5%) and viability (53.5%) based on WHO criteria. However, fertile men had a low mean percentage of normal sperm morphology (20.0%), although they were normally distributed. Cigarette smoking was associated with significantly lower semen volumes even after adjusting for alcohol consumption. The sperm parameters (i.e. volume, density, motility, viability and normal morphology) were not significantly associated with ethnic differences. The WHO criterion for normal sperm morphology is too stringent, and should be adopted with caution. Normal sperm morphology is but one of many parameters for assessment of fertility. Social alcohol consumption, cigarette smoking, and 'recent fever' did not appear to affect sperm quality in this group of fertile men.      

A new test for the assessment of sperm- zona pellucida penetration: relationship with results of other sperm tests and fertilization in vitro

The spermatozoa of some patients attending for in-vitro fertilization(IVF) fail to penetrate the zona pellucida in vitro. A testhas been devised to identify these cases. It is based on thenumber of spermatozoa penetrating into the zona pellucida, whichwere counted after removing spermatozoa bound to the zona surfaceby vigorous aspiration of each oocyte through a narrow gauge(120 µm) glass pipette. The oocytes were collected from197 patients undergoing IVF treatment with their own gametes;79 with no oocytes fertilized and 118 with some oocytes fertilized.Sperm motility, morphology and DNA normality (acridine orangestain) were also measured. The relationships between sperm testresults and IVF rate were examined by logistic regression. Theproportions of penetrated zonae, normal sperm morphology andnormal DNA were the most significant factors related to IVFrate in the whole group. Also, in patients with 30 spermatozoabound per zona pellucida or with normal sperm morphology 30%,the proportion of penetrated zonae and normal DNA were mostsignificant. Oocytes from 42 patients who had zero fertilizationand low sperm-zona binding (average, 2.2 spermatozoa/zona pellucida)were re-incubated with normal donor spermatozoa: large numbersof spermatozoa bound (average, 88 spermatozoa/zona pellucida)and each zona was penetrated by at least one spermatozoon. Inconclusion, the percentage of zonae penetrated was the variablemost significantly correlated with IVF rate. Penetration ofthe zona was also strongly related to fertilization rates inpatients without defects of sperm morphology and sperm-zonabinding. In patients where all zonae were penetrated, poor fertilizationmay be due to sperm morphology and DNA abnormalities. Failureof sperm-zona binding and penetration in vitro in patients withfailure of fertilization was mainly due to sperm defects andnot oocyte defects