影响人类精子耐冻性的蛋白质分析

目的:利用比较蛋白质组学技术研究影响人类精子耐冻性的蛋白质。方法:将10例供精志愿者的31份精液标本依据冷冻复苏率的不同分为高冷冻复苏率组和低冷冻复苏率组,应用二维电泳技术和质谱分析鉴定技术寻找两组标本的精子蛋白和精浆蛋白在表达量上的差异。结果:在精子蛋白和精浆蛋白中共发现了22个差异表达蛋白。其中精浆蛋白12个,分别为细胞角蛋白1、神经元导向因子3、Rho2样蛋白、Krueppel样因子10、FERM域蛋白8、GMP合成酶、鸟嘌呤结合蛋白G、溶质载体家族12、ATP结合盒亚家族D1、6-磷酸果糖-2-激酶、肌钙蛋白C、肌球蛋白轻链;精子蛋白9个,分别为卷曲螺旋结构域蛋白113、RAF原癌基因丝氨酸/苏氨酸蛋白激酶、Ras相关蛋白Rab-31、α微管蛋白3C/D、锌指蛋白879、胆固醇26-羟化酶、DNA修复和重组蛋白、细胞角蛋白73、钙磷蛋白;精浆和精子共有蛋白1个,为细胞周期依赖性蛋白激酶1。文献调研发现这些差异蛋白主要与精子的能量代谢、成熟、运动、DNA修复等有关。结论:鉴定了多种可能影响人类精子耐冻性的蛋白质,为以后从分子水平上揭示精子冷冻复苏原理提供了依据。     

人类精液冷冻前后精子运动变化的计算机辅助分析

目的 :利用计算机辅助精液分析 (CASA)系统分析冷冻前后人类活动精子的运动参数 ,并观察相关参数的变化规律。　方法 :2 38份精液样本在冷冻前和复苏后均采用CASA分析其运动参数。　结果 :复苏后精子活动率下降 ;除精子鞭打频率 (BCF)外 ,冷冻前与复苏后全部精子运动参数呈显著正相关且差异存在显著性 ;平均路径速度 (VAP)、直线速度 (VSL)、曲线速度 (VCL)下降 ,精子头侧摆幅 (ALH)下降 ,但直线性 (LIN)、前向性 (STR)升高。　结论 :CASA系统可对冻融前后精子运动参数进行客观细致的分析 ,有一定的临床应用价值。     

部分型圆头精子症患者精液常规参数及精子形态分析

目的:分析部分型圆头精子症患者精液常规参数及精子形态。方法:选取2013年1月至2016年5月100例部分型圆头精子症患者作为病例组,对照组为非圆头精子症的不育患者180例。根据精液中圆头精子所占比例不同将病例组分成5组:1组(25%~40%)、2组(41%~55%)、3组(56%~70%)、4组(71%~85%)、5组(86%~99%),根据WHO《人类精液检查与处理实验室手册》第5版标准对其进行精液常规分析,包括精子浓度、活动率、前向运动精子百分率,及对不同畸形精子进行计数并计算畸形精子指数(TZI)及精子畸形指数(SDI)。结果:病例组与对照组的精子活动率[(35.76±24.88)%vs(62.03±10.20)%]、前向运动精子百分率[(26.11±20.39)%vs(45.62±6.87)%]、正常形态精子百分率[(1.45±1.45)%vs(5.98±2.21)%]和SDI(1.33±0.11 vs1.27±0.57)相比差异均有统计学意义(P均0.01),两组年龄[(29.82±4.90)岁vs(30.33±3.59)岁]、精子浓度[(46.01±40.38)×10~6/ml vs(54.00±25.15)×10~6/ml]和TZI(1.35±0.11 vs 1.34±0.54)比较差异均无统计学意义(P均0.05);病例组各组精子活动率、前向运动精子百分率、正常形态精子百分率、TZI和SDI均有统计学差异(P均0.01),而年龄和精子浓度均无统计学差异(P均0.05)。随着圆头精子比例的升高,精子头部形态由异质性偏向于均一性。结论:精液中不同比例的圆头精子与精液常规参数和形态学指标密切相关,可对不育患者辅助生殖技术的选择和受精率的预测起一定的提示作用。     

Macro精子计算盘测定精子运动速度及与精液常规参数的比较

Ｍａｃｒｏ精子计算盘测定精子运动速度及与精液常规参数的比较高正洪关键词Ｍａｃｒｏ精子计算盘，精子运动速度，精液常规参数中图法分类号Ｒ４４６．１９精液常规分析是评价男性生育力的一个重要指标。用Ｍａｋｌｅｒ计算盘检查精液已被ＷＨＯ确定为精液检查的标准方法...     

人类精子库自体精液冻存失败原因分析及应对策略

目的:分析人类精子库意愿自精保存者保存精液失败的原因,针对性提出应对策略,尽可能提高自体精液冻存成功率,同时也为人类精子库自体精液冻存业务拓展提供建议.方法:回顾性分析广东省人类精子库2016年1月1日至2019年12月31日1156例意愿自精保存者的基本资料、存精理由和存精失败的原因等信息.结果:1156例意愿自精保...     

213例不育男性抗精子抗体与精液有关参数的分析

我们检测了９００多例男性不育症患者的血清，精浆抗精子抗体和精液各项指标，对其中２１３例ＡｓＡｂ阳性患者的精液有关参数与ＡｓＡｂ的关系进行了分析，并与生育男性组作了比较。结果表明，ＡｓＡｂ阳性患者的精子密度精子活率，精子顶体完整率，精浆免疫抑制物及α－糖苷酶均明显人氏于生育组，不育男性ＡｓＡｂ阳性组ＳＰＩＭ，阳性率也明显高天ＡｓＡｂ阳性率也明显高于ＡｓＡｂ阴性组。此外，血清与精浆ＡｓＡｂ同为阳性组的     

计算机辅助精液分析男性不育患者精子运动参数与精子活力的相关性

目的探讨计算机辅助精液分析精子运动参数在评价男性不育患者精子活力中的价值。方法按《WHO人类精液及精子-宫颈黏液相互作用实验检验手册》标准,采用国产WLJY-9000伟力彩色精子质量检测系统对276例男性不育患者的精液进行平均直线运动速度、平均曲线运动速度、运动的前向性、运动的直线性、运动的摆动性、平均路径速度、精子活力及分级等进行检测并分析其相关性。结果276名男性不育患者的平均精子活力为（48.93±19.10）%,分级为A级（32.11±17.25）%、B级（17.03±8.91）%、C级（10.14±5.99）%。平均直线运动速度、平均曲线运动速度、运动的前向性、运动的直线性、运动的摆动性、平均路径速度与精子活力的相关系数分别为0.60（P〈0.01）、0.59（P〈0.01）、0.51（P〈0.01）、0.55（P〈0.01）、0.52（P〈0.01）、0.67（P〈0.01）。结论计算机辅助精液分析精子运动参数平均直线运动速度、平均曲线运动速度、平均路径速度是反映精子活力的有效指标,精子运动参数对男性不育的诊断和生育能力的评估具有实用意义。     

精液粘度与精液分析参数、解脲支原体感染和抗精子抗体关系分析

目的探讨精液粘度增高导致男性不育的机理。方法共4337例不育门诊就诊者,分为精液粘度增高和正常组,观察粘度与主要精液常规参数、UU感染率和AsAb阳性率关系。结果精液粘度增高率为65.02%。粘度增高组精子活动率、a,b级活力精子率显著低于粘度正常组(P<0.05,P<0.001);畸形精子率、液化时间均明显高于粘度正常组(P<0.001);两组精液量、精子密度和精液pH比较无显著性差异(P>0.05)。精液白细胞>5个/HP组粘度增高率明显高于白细胞<5个/HP组(P<0.001)。粘度增高组精浆AsAb阳性率和精液UU阳性率均明显高于粘度正常组(P<0.001)。结论精液粘度可影响精液参数,并与精液白细胞数、精浆AsAb和UU感染有关。     

精液中凋亡精子与精液分析参数的相关性研究

凋亡是许多哺乳类物种正常生精过程中调节生殖细胞增殖的一种机制。但发生在精子水平的凋亡是否与精液中精子的数量和质量有关尚不清楚。磷脂酰丝氨酸（PS）是丝氨酸以酯键与磷脂酸的磷酸基相连的一种磷脂，在细胞凋亡的早期阶段，细胞膜的脂质不对称性丧失，位于细胞质膜内的PS转移到膜外。     

213例不育男性抗精子抗体与精液有关参数的分析

我们检测了９００多例男性不育症患者的血清、精浆抗精子抗体（ＡｓＡｂ）和精液各项指标，对其中２１３例ＡｓＡｂ阳性患者的精液有关参数与ＡｓＡｂ的关系进行了分析，并与生育男性组作了比较。结果表明，ＡｓＡｂ阳性患者的精子密度、精子活率、精子顶体完整率、精浆免疫抑制物（ＳＰＩＭ）及α－糖苷酶均明显低于生育组（Ｐ＜０．０１），不育男性ＡｓＡｂ阳性组ＳＰＩＭ阳性率也明显高于ＡｓＡｂ阴性组（Ｐ＜０．０１）。此外，血清与精浆ＡｓＡｂ同为阳性组的精子活率明显低于单纯血清或精浆ＡｓＡｂ阳性组（Ｐ＜０．０１）。     

2343例男科就诊患者精子动态参数分析

目的探讨精子动态参数在评价男性生育能力方面的应用价值。方法严格按照WHO技术规范，用计算机辅助精液分析（CASA）系统对本院2343例男科门诊就诊的患者精液进行常规指标和精子动态参数分析。结果①正常精液组和异常精液组各项精子动态参数比较，均存在统计学差异；②随着精子存活率的下降，曲线速度（VCL）、直线速度（VSL）、平均路径速度（VAP）、平均移动角度（MAD）、侧摆幅度（ALH）、前向性（STR）下降明显，鞭打频率（BCF）呈上升趋势，而直线性（LIN）、摆动性（WOB）差异无显著性；③与密度活力正常组比较：少精组除LIN、STR有所增加，MAD、BCF有所下降，差别有统计学意义外，其余五项参数差别无统计学意义；弱精组除MAD无统计学差异外，其余八项参数均有统计学差异；少弱精组除MAD和ALH无统计学差异外，其余七项参数均有统计学差异。结论精子动态参数是评估精液质量的重要指标，为男性不育的诊断、治疗和研究提供了充分的依据。     

Human sperm DNA integrity in normal and abnormal semen samples and its correlation with sperm characteristics

Reports indicate an increase in the incidence of DNA fragmentation in male factor infertility and its role in the outcome of assisted reproductive techniques (ART). However, reports are conflicting between the relationships of sperm DNA integrity with conventional semen parameters. We examined the relationship between conventional sperm parameters and DNA integrity using acridine orange (AO) test. The study included 373 patients and 28 fertile volunteers. DNA normality was compared with semen parameters between the patient and donor populations. Significant correlations were noted between DNA normality and sperm concentration ( r  = 0.18, P  = 0.000), motility ( r =  0.21, P  = 0.0001), rapid motility (0.19, P  = 0.000), normal morphology by World Health Organization ( r =  0.15, P  = 0.019) and head defects ( r =  −0.15, P  = 0.023). A significant difference was noted in AO levels between donors and patients with asthenozoospermia ( P  = 0.002) and oligoasthenozoospermia ( P  = 0.001). A significant difference in DNA integrity was noted in samples having <30% and >30% normal morphology. A wide range of % DNA normality was observed in the patient group. Sperm assessment for DNA status using AO is reliable and shows good correlation with sperm count, motility and morphology. Assessment of sperm DNA status with AO staining may be helpful prior to ART.     

Relationship between age and semen parameters in men with normal sperm concentration: analysis of 6022 semen samples

This study evaluates retrospectively the relationship between age and semen parameters among men with normal sperm concentration. It was based on computerized data and performed in an Academic Fertility and IVF Unit. Six thousand and twenty-two semen samples with sperm concentrations of >or=20 x 10(6) ml(-1) were examined according to WHO criteria and analysed in relation to patients' age. For each age group, mean values +/- SD of semen volume, sperm concentration, percentage of motile spermatozoa, normal morphology, acrosome index, total sperm count/ejaculate, total motile sperm count/ejaculate and sexual abstinence duration were examined. A peak semen volume of 3.51 +/- 1.76 ml(-1) was observed at age >or=30 to <35 years and a lowest volume of 2.21 +/- 1.23 ml(-1) was observed at age >or=55 years (P<0.05). Sperm motility was found to be inversely related to age with peak motility of 44.39 +/- 20.69% at age <25 years and lowest motility of 24.76 +/- 18.27% at age >or=55 years (P<0.05). A reduction of 54% was observed for total motile sperm, between values of 103.34 +/- 107 x 10(6) at age >or=30 to <35 years and 46.68 +/- 53.73 x 10(6) (P<0.05) at age >55 years. A statistically significant and inverse relationship was observed between semen volume, sperm quality and patient age, in spite of prolonged sexual abstinence duration. Top sperm parameters were observed at age >or=30 to <35 years, while the most significant reduction in sperm parameters occurred after the age of 55 years.     

The mean of sperm parameters in semen donations from the same donor. An important prognostic factor in insemination

We analysed 12,100 consecutive cycles of artificial insemination by donor spermatozoa in 1901 infertile couples. In our analysis, particular attention was given to finding an appropriate way of taking into account the respective effects of female and male factors on the pregnancy success rate and the level at which these factors act (cycle vs. woman and donation vs. donor). A total of 1213 pregnancies occurred. The pregnancy rate per cycle was lower as the age of the woman increased (p < 0.0001) and varied with the type of infertility: fecundity was higher (p = 0.03) in the case of azoospermia than of severe oligozoospermia. After taking into account these factors, significant unexplained variation in likelihood to conceive remained. A part of this heterogeneity was shown to be due to variation in fecundability between semen donors. In order to explain this heterogeneity between donors, compositional covariates were used, particularly the mean of results of the semen analysis performed for donations from the same donor. For each semen characteristic, the overall mean of the different donations of a donor was an important predictive factor of successful insemination: after taking into account all of the other factors, the odds ratios for an increase of 50 x 10(6)/mL spermatozoa, of a 20% increase in sperm motility and of a 2 point increase in the post-thaw quality index, were, respectively, 1.13, 1.37 and 1.56. After adjustment for these factors, the specific characteristics of each semen donation were no longer significantly predictive of successful insemination. This observation has a biological interpretation: sperm with low parameters but produced by a normally fertile man can have a satisfactory success rate.     

上海市人类精子库捐精现状及临床应用回顾性分析

目的 探讨上海市人类精子库捐精现状与临床应用,为中国精子捐献制度的完善提供理论依据.方法 对上海市人类精子库自2003年3月至2010年12月间所有初次捐精者与合格捐精者的信息回顾性分析,研究内容包括:(1)捐精者筛查不合格原因;(2)合格捐精者脱失率及原因;(3)捐精动机;(4)精液标本冷冻前后参数;(5)捐献者精子用于辅助生殖治疗的结果.结果 1.研究期间,共有12 858名志愿者接受捐精筛查,只有2 083名(16.2％)入选.落选的主要原因为:(1)精液参数未达到标准;(2)性传播疾病检测阳性结果.另外,约4.9％的合格捐精者脱失,其主要原因为(1)捐精期间多次精液检测未达到标准,丧失信心;(2)因免费医学检查而加入;(3)迁居外地.2.我库共冻存精液3 1 056份,冷冻前后平均密度为(98.7±25.3)×106和(57.2±26.5)×106,前向运动精子比率为(62.1±15.3)％和(42.6±13.5)％.3.对外供精14 181份,其中AID周期数为7 341,妊娠率为20.13％,新生儿出生数为723人;体外受精-胚胎移植(IVF-ET)周期数为1521,妊娠率为38.66％,新生儿出生数为410人,未出现同一供精者使5名以上妇女受孕的情况.结论 我国精子库已进入成熟发展阶段,遵循国家政策法规的严格管理.目前存在的问题是供精严重不足,这有赖于政策进一步完善.     

SCSA和SCD检测精子DNA完整性结果比较及与精子质量参数的相关性分析

目的:近年来精子DNA碎片预测男性生育受到广泛关注,其检测方法较多,但方法学的比较研究很少,本文旨在比较精子DNA完整性常用检测方法之间的差异,并分析DNA碎片与精子质量的相关性。方法:选择108例精液样本,采用精子染色质结构分析试验(SCSA)和精子染色体扩散实验(SCD)对样本分别进行精子DNA碎片检测分析,对两种方法的结果进行比较,并与精液常规、精子形态以及患者年龄进行相关性分析。结果:两种方法检测的精子DNA碎片化指数(DFI)存在显著的一致性(P0.01),DFI与精子活动率、前向运动精子百分率以及正常形态精子百分率呈显著负相关(P0.01),与畸形精子指数呈显著正相关(P_(SCSA)0.01,P_(SCD)0.05),SCSA得出的DFI与年龄呈显著正相关(P0.01),而SCD法的DFI并未发现类似的现象。结论:两种方法检测的结果对精子质量均有较好的预测价值,精子DFI作为一个检测指标,对男性生育力预测有一定的临床价值。但不同方法之间的检测结果差异较大,检测方法的标准化有待进一步研究。     

Sperm indexes obtained using computer-assisted morphometry provide a forecast of the freezability of canine sperm

Sperm morphometric indexes obtained after principal component analysis were used to investigate its value as diagnostic tests for freezability. Semen from six dogs was frozen-thawed following a standard protocol. Before freezing, computer-assisted analysis of sperm morphometry (CASMA) was performed. The principal component analysis (a statistical technique for simplifying a dataset, by reducing multidimensional datasets to lower dimensions for analysis) performed in the data obtained after the CASMA analysis gave four morphometric indexes. After thawing, ejaculates were evaluated for early changes in sperm membranes using the combination of two fluorescent probes, YO-PRO-1 and ethidium homodimer and flow cytometry. Significant differences in the percentages of intact membranes post-thaw were observed among dogs (p < 0.01). Significant non-parametric correlations were found between index 3 and the percentage of intact membranes after thawing (R = 0.432 p < 0.05). Receiving operating system curves demonstrate a good diagnostic value for indexes 2 and 3 in the prediction of freezability, with areas under the curve of 0.798 and 0.786, respectively.     

Human sperm acrosin activity with relation to semen parameters and acrosomal ultrastructure

Summary. It was suggested that although not related to the standard semen parameters the level of the acrosin enzyme system is related to the fertility potential in men. Recently a simple clinical assay for total acrosin level was recommended for routine semen analysis. The improved clinical assay was analysed on the freshly liquified semen of 198 Astheno-teratozoospermic men and compared with the routine semen parameters including biochemical data and the ultrastructure of the acrosome. Only the sum of the per cent of live, motile, and normal-shaped spermatozoa had positive significant and reasonably high correlation with the acrosin level ( r = 0.382, P < 0.0001). Each characteristic exhibits significant but low (< 0.35) correlation. Similarly negative significant and reasonably high correlation was obtained between the acrosin level and the sum of the principle acrosomal malformations observed by TEM ( r = 0.396, P < 0.0001) while lower negative correlation was found only with agenesis or loss of the acrosome. Acrosin levels below 8.1 μIU 10⁻⁶ cells were obtained in 4 specimens with above 80% round-form associated with more than 95% of agenesis of the acrosome. The possible significance of the low correlation obtained between the acrosin levels and seminal plasma zinc levels, malformations in the acrosomal equatorial region, and the presence of white blood cells is also discussed. We concluded that the acrosin activity reflects an aspect of male fertility which is not diagnosed by the routine semen analysis or by the ultrastructure of the acrosome, and is therefore a useful diagnostic sperm parameter.     

Sperm decondensation and semen parameters: utilization of a simple staining technique for the evaluation of human sperm decondensation

Summary. The ability of spermatozoa to fertilize an oocyte depends on a sequence of events ending ultimately in the decondensation of the sperm chromatin on penetration of the oocyte. Knowledge of what percentage of sperm decondenses is useful, especially in patients where other functional tests and sperm quality fail to explain the reported poor in vitro fertilization (IVF) rates. The objective of this study was (1) to compare sperm decondensation induced by either SDS/EDTA or heparin with semen parameters (volume, concentration, motility and morphology), and (2) to evaluate the use of a simplified staining technique (Diff Quik^R [DQ]) in comparison with the standard phase contrast method (Rose Bengal-[RB]). Randomly selected semen samples from 31 men attending an assisted reproductive programme were analysed for basic semen parameters and decondensation with SDS/EDTA and heparin. Two staining methods for the evaluation of decondensation were compared (phase contrast microscopy after Rose Bengal [RB] staining and light microscopy after Diff Quik^R (DQ) staining). Moderate and grossly swollen sperm heads were recorded. Semen samples included both fertile and unfertile semen parameters. Sperm decondensation results showed poor to moderate correlations with semen parameters. The SDS/EDTA (DQ) (moderate forms) showed a significant negative correlation (r = -0.46) with seminal volume and and a significant positive correlation (r = 0.41) with normal sperm morphology. The heparin (DQ) (moderate forms) decondensation showed a significant positive correlation with motility (r = 0.61) and sperm concentration (r = 0.43). The DQ method was preferred over the RB method due to its optical and storage advantage. Sperm decondensation by SDS/EDTA and heparin have limited use in the IVF laboratory as they correlate poorly with semen parameters. Future studies should investigate the use of an ooplasmic factor similar to nucleoplasmin in Xenopus laevis egg.     

深圳地区与其他地区男科就诊患者精液质量比较分析

目的 了解深圳地区就诊男科患者的精液质量特征.方法 对北京大学深圳医院泌尿男科及辅助生育科室就诊病人的6 964份精液标本检测结果进行统计分析,并将结果与成都、遵义及西班牙男科门诊就诊病人精液质量进行比较.分析指标包括精液量、密度、活动率、活力(A+B)精子百分比等.结果 在6964份精液中,正常的精液占19.74%,弱精子症占50.72%,少精子症占24.11%,无精子症占2.77%,其他指标异常占2.66%.各项指标均数:精液量(3.04±1.05)ml,精子密度(50.73±42.11)×106/ml,精子总数(148.03±127.05)×106/ml,精子活动率(50.06±17.90)%,a级精子(17.98±11.75)%,(a+b)级精子(32.80±15.14)%.对比成都、遵义地区的相关数据,精子密度下降、精子活动率下降、精子活力下降等精液异常的百分比较高,差异有统计学意义,而无精子症百分比差异有统计学意义.对比西班牙相关数据,(a+b)级精子百分比差异无统计学意义(P>0.05);精子密度、(a+b)级精子数目高于西班牙(P<0.01);精液量、禁欲时间、精子总数低于西班牙指标,差异有统计学意义(P<0.01).结论 深圳地区就诊男科患者精液质量低于成都地区及遵义地区水平,与西班牙比较精液质量无明显下降.