雷达作业人员精子染色质结构分析

目的 :评价雷达辐射对作业人员精子DNA的损伤。　方法 :对高暴露组 (n =88)、低暴露组 (n =14 3)、对照组 (n =39)进行精液常规分析和精子染色质结构分析 (SCSA)。　结果 :高暴露组较低暴露组、对照组精子活力和存活率下降 ,畸形精子率增加 ;高暴露组主群以外细胞数值 (COMPα)较低暴露组、对照组升高 ,且其具有中生育力潜能。变量的多因素分析表明 ,日工作时间为畸形精子率的危险因素 ,禁欲时间为SCSA参数SDαt、COMPα的危险因素。　结论 :雷达辐射对男性生殖系统有不良影响 ,采取防护措施很重要。     

少弱畸精子综合征治疗前后精子染色质流式细胞术分析

目的:观察少弱畸精子综合征(OAT)患者采用中西医结合治疗前后精子染色质的变化。方法:对60例OAT患者给予中西医结合治疗3个月,留取治疗前后精液标本,进行吖啶橙(AO)标记,流式细胞术分析以及激光共聚焦显微镜观察绿、黄和红色荧光精子百分率,同时进行常规分析。结果:治疗前后流式细胞术提示主群精子变化存在显著差异(P<0.01)、αt和COMPαt变化存在显著差异(P<0.05)。激光共聚焦显微镜观察时,绿色与红色两种荧光有显著差异(P<0.05),而黄色荧光无明显差异(P>0.05)。计算机辅助精子分析结果:精子密度、活动力、活动率和畸形率等均有变化。除直线运动精子密度无显著差异外(P>0.05),密度、活动率、活动力均有显著差异(P<0.05)。结论:中西医结合治疗OAT综合征可以改善精子染色质。综合流式细胞术、CASA、精子AO染色激光共聚焦显微镜观察等3种检测手段更有助于疗效判断。     

精子染色质结构分析与精液参数的相关性研究

目的 :研究精子染色质结构分析 (SCSA)与精液参数的相关性 ,探讨评估精液质量的可靠方法。　方法 :将5 11份精液标本流式细胞术SCSA结果与精液常规分析结果进行多参数的相关性分析。　结果 :与变性精子百分数(COMPαt)呈低度正相关 (r在 0 .10～ 0 .3 0 )的参数是粘度、本次射精间隔时间、精子畸形率、c级精子密度 ;呈低度负相关的参数 (r在 - 0 .3 0～ - 0 .10 )是曲线速度、直线速度、平均路径速度 ;呈中度正相关 (r在 0 .3 0～ 0 .70 )的参数是精子密度、d级精子率和d级精子密度 ;呈中度负相关 (r在 - 0 .70～ - 0 .3 0 )的参数是平均移动角度、a级精子密度和精子存活率。　结论 :流式细胞术能简单、快速、准确地评估损伤和变性精子的百分数 ,其结果 (COMPαt)与精液参数部分相关 ,并不是完全独立于精液常规检查的指标。SCSA对评估生育力具有重要意义     

精索静脉曲张对精子染色质结构及运动能力的影响

目的:探讨精索静脉曲张对精子染色质结构及运动能力的影响。方法:对精索静脉曲张组(无慢性疾病及生殖系统病史者,n=74)和对照组(n=89)进行精子染色质结构分析(SCSA)和精液常规分析(精液量、粘稠度、pH值、精子顶体完整率、精子存活率、畸形率、精子密度、活动率及各运动参数)。结果:精索静脉曲张组精子密度、a+b级精子百分率和精子存活率[分别为(41.4±38.7)×106/ml,(31.7±16.9)%,(62.8±22.2)%]显著低于对照组[分别为(80.9±63.1)×106/ml,(46.8±20.5)%,(77.2±17.5)%],P均<0.05;精子运动参数VCL、VSL、VAP精索静脉曲张组分别为(37.4±12.5)、(23.4±7.8)、(26.5±8.2)μm/s,对照组分别为(42.4±10.7)、(27.3±7.3)、(30.7±7.8)μm/s,精索静脉曲张组与对照组相比显著降低(P均<0.05),而MAD显著增加(P<0.01),SCSACOMPαt值精索静脉曲张组(为23.2±16.2)明显高于对照组(为14.1±11.8)(P<0.05)。结论:精索静脉曲张引起精子DNA损伤和精子运动能力的改变,可能是导致男性不育的主要原因之一。     

精子染色质扩散实验检测精索静脉曲张及不明原因不育患者精子DNA碎片

目的了解精索静脉曲张(VC)及不明原因不育患者精子DNA碎片的发生比例。方法改进的精子染色质扩散(SCD)实验分析精子DNA碎片。检测VC不育患者39例,不明原因不育患者57例。以生育健康成年男性32例为对照组。结果VC不育患者SCD小光晕和无光晕精子(精子DNA碎片)比值平均为(36.6±18.9)%,VC不育组明显高于对照组(12.1±5.2)%(P<0.001),而大光晕和中光晕精子比值VC不育组明显低于对照组(P<0.01);不明原因不育患者精子DNA碎片比值平均为(26.8±10.2)%,与对照组[(12.1±5.2)%]比较有显著性差异(P<0.001)。结论SCD实验表明,VC及不明原因不育患者精子DNA碎片比值增高。     

吸烟对精子染色质结构完整性影响的探讨

目的:探讨吸烟对精子染色质结构完整性的影响。方法:784例男性不育患者从病例库中选取,根据吸烟与否及每日吸烟支数(≤10、11~19、≥20)和吸烟年限(≤5、6~9、≥10)进行分组,比较各组患者精液常规检测参数及精子染色质结构受损和精子核成熟情况。精子染色质结构完整性采用流式细胞仪检测,DNA损伤程度和不成熟精子指数分别以DNA损伤指数(DFI)和高DNA着色性(HDS)来表示。结果:吸烟≥20支/日和吸烟年限≥10年的患者组精液量、前向运动精子比例低于其他组而精子畸形率高于其他组(P<0.05);吸烟组男性的DFI和HDS值均升高(P<0.05);HDS与前向运动精子比例呈负相关(r=-0.18,P<0.05),DFI与HDS均与精子畸形率呈正相关(r=0.31与r=0.39,P均<0.05)。结论:日吸烟量≥20支或吸烟年限≥10年对男性的精液量、前向运动精子比例和精子畸形率都有影响,吸烟影响男性精子DNA完整性和精子核成熟。     

精子碎片实验室检测与临床应用专家共识——基于流式细胞精子染色质结构分析

在过去的几十年里,精子DNA碎片(SDF)检测是一项经受住时间考验的精子功能研究技术,在全世界的男科实验室中日益普及.各种体外和体内研究都支持精子DNA完整性在受精、早期胚胎发育、植入和妊娠中的重要性[1-2].对于接受辅助生殖技术的患者,SDF已经成为一种新的风险分层生物标志物[3].1980年,第1次SDF分析,引...     

精子染色质结构分析预测夫精宫腔内人工授精结局的临床研究

目的:分析精子DNA损伤与宫腔内人工授精(intrauterine insemination,IUI)妊娠率的关系。方法:482例行IUI治疗的患者,常规精液分析,并采用精子染色质结构分析法(SCSA)计算DNA碎片指数(DNA frag-mentation index,DFI),观察DFI与IUI妊娠率的关系。结果:482例患者中DFI>25%的患者为95例,5例临床妊娠,妊娠率为5.26%;DFI≤25%的患者为387例,59例临床妊娠,妊娠率为15.25%。DFI>25%的患者,其生化妊娠率和临床妊娠率只有DFI≤25%的患者的0.37(95%CI:0.14～0.96)和0.38(95%CI:0.16～0.97)倍。54例患者前后2个周期DFI值分别为(15.05±7.98)%与(17.25±12.18)%,差异无统计学意义(P>0.05)。精子DFI与精子浓度、精子活力、前向运动精子总数呈显著负相关(P<0.01)。结论:SCSA检测的DFI参数是较稳定的指标,DFI和精子浓度、精子活力具有一定相关性。DFI>25%的患者IUI妊娠率明显低于DFI≤25%的患者。SCSA检测的DFI参数是一种较为理想的预测IUI临床妊娠率的指标。     

精液体外处理对精子染色质完整性的影响

<正>精子染色质完整性损伤可以增加不育和遗传缺陷风险,精子染色质完整性损伤的因素及其保护机制已成为辅助生殖领域研究的热点之一。精液体外处理技术是辅助生殖技术(ART)常规操作,研究表明,经体外优选处理后,精子浓度、活力、形态等可很大程度改善[1],然而其对精子染色质完整性的影响目前尚存在争议。体外处理精液,去除精浆的同时     

精子染色质完整性对IVF/ICSI临床结局的预测价值

目的:通过分析精子染色质完整性与体外受精胚胎移植(IVF-ET)、卵细胞胞质内单精子注射(ICSI)结局之间的关系,探讨精子染色质完整性检测在辅助生殖技术(ART)中的预测价值。方法:采用精子染色质结构分析试验(SCSA)方法检测187个ART周期中精子的染色质完整性,以精子DNA损伤指数(DFI)为参数,分为高DFI组(DFI≥30%)和低DFI组(DFI<30%),两组中根据采用不同的体外受精方式分为IVF亚组和ICSI亚组。分别比较高DFI组、低DFI组中IVF亚组和ICSI亚组间受精率、卵裂率、D2天胚胎质量、D3天胚胎质量、临床妊娠率差异性。结果:在高DFI组中,行ICSI治疗的夫妇临床妊娠率显著高于行IVF治疗的夫妇,具有统计学意义;在行IVF治疗的夫妇中,比较高DFI组与低DFI组的各临床结局,未见统计学差异;在行ICSI夫妇中,比较高DFI组与低DFI组的各临床结局,未见统计学差异。结论:精子染色质完整性影响辅助生育技术的结局,在选择辅助生殖技术方案时,可作为常规精液检查的一个补充。     

Sperm chromatin structure assay （SCSA）： a tool in diagnosis and treatment of infertility

Diagnosis of male infertility has mainly been based on the World Health Organization (WHO) manual-based semen parameter's concentration, motility and morphology. It has, however, become apparent that none of these parameters are reliable markers for evaluation of the fertility potential of a couple. A search for better markers has led to an increased focus on sperm chromatin integrity testing in fertility work-up and assisted reproductive techniques. During the last couple of decades, numerous sperm DNA integrity tests have been developed. These are claimed to be characterized by a lower intraindividual variation, less intralaboratory and interlaboratory variation and thus less subjective than the conventional sperm analysis. However, not all the sperm chromatin integrity tests have yet been shown to be of clinical value. So far, the test that has been found to have the most stable clinical threshold values in relation to fertility is the sperm chromatin structure assay (SCSA), a flow cytometric test that measures the susceptibility of sperm DNA to acid-induced DNA denaturation in situ. Sperm DNA fragmentation as measured by SCSA has shown to be an independent predictor of successful pregnancy in first pregnancy planners as well as in couples undergoing intrauterine insemination, and can be used as a tool in investigation, counseling and treatment of involuntary childlessness. More conflicting data exist regarding the role of sperm DNA fragmentation in relation to fertilization, pre-embryo development and pregnancy outcome in in vitro fertilization and intracytoplasmic sperm injection (ICSI).     

Relationship between sperm quality and chromatin condensation measured by sperm DNA fluorescence using flow cytometry

DNA flow cytometry of sperm from 100 randomly chosen men undergoing fertility investigation revealed a general association between reduced sperm quality, as judged by conventional parameters, and the appearance of sperm with lower degrees of chromatin condensation in the ejaculate as measured by DNA fluorescence intensity. Chromatin hypocondensation, as measured by increased fluorescence, was manifested to different degrees in different samples. In many cases of more extreme sperm pathology, such as oligoasthenoteratozoospermia (OAT), the whole population of spermatozoa appeared to be affected. Significant numbers of hypercondensed spermatozoa were present in both normozoospermic men and men with different degrees of disturbance in sperm quality. All of the different parameters of sperm quality could be correlated significantly with certain of the flow parameters, although not one in particular could be used to predict deviations from the normal flow profile. In several asthenoteratozoospermic men and a small proportion of men with OAT, the DNA profiles were normal, implying that in these cases the disturbance may not be so fundamental. The presence of leucocytes in the ejaculate was associated with a general increase in the preponderance of hypocondensed subpopulations of spermatozoa in men with OAT as well as in normozoospermic subjects, emphasizing the effect of inflammatory conditions in the reproductive tract on sperm quality.     

Sperm chromatin structure assay as an independent predictor of fertility in vivo: a case–control study

Standard sperm parameters have a limited power for prediction of the chance of natural conception. Recent studies have indicated that the sperm chromatin structure assay (SCSA) DNA fragmentation index (DFI), a measure for the fraction of sperms with DNA damage, is associated with fertility in vivo . The aim of this study was to evaluate the value of this parameter for prediction of infertility. One hundred and twenty-seven men from infertile couples with no known female factor and 137 men with proven fertility were included. Semen analysis was performed as recommended by the WHO. DFI was assessed using SCSA. Logistic binary regression was used to compute the odds ratios (OR) for infertility. As compared with men with a DFI <10%, men with a DFI between 10% and 20% had an increased risk for infertility (OR 2.5, 95% CI: 1.0–6.1). This was also true for men with a DFI >20% (OR 8.4; 95% CI: 3.0–23). In men with normal standard semen parameters (sperm concentration, motility and morphology) the OR for infertility was increased with DFI >20% (OR 5.1, 95% CI: 1.2–23), whereas if one of the standard semen parameters was abnormal, the OR for infertility was increased already at DFI above 10% (OR 16, 95% CI: 4.2–60). We conclude that SCSA DFI adds to the value of semen analysis in prediction of the chance of natural conception.     

Sperm chromatin structure assay results after swim-up are related only to embryo quality but not to fertilization and pregnancy rates following IVF

The aim of this study was to investigate whether the sperm chromatin structure assay (SCSA) results after swim-up are related to fertilization rates, embryo quality and pregnancy rates following in vitro fertilization (IVF). A total of 223 couples undergoing IVF in our hospital from October 2008 to September 2009 were included in this study. Data on the IVF process and sperm chromatin structure assay results were collected. Fertilization rate, embryo quality and IVF success rates of different DNA fragmentation index (DFI) subgroups and high DNA stainability (HDS) subgroups were compared. There were no significant differences in fertilization rate, clinical pregnancy or delivery rates between the DFI and HDS subgroups. However, the group with abnormal DFI had a lower good embryo rate. So, we concluded that the SCSA variables, either DFI or HDS after swim-up preparation, were not valuable in predicting fertilization failure or pregnancy rate, but an abnormal DFI meant a lower good embryo rate following IVF.     

Clinical correlates of the biological variation of sperm DNA fragmentation in infertile men attending an andrology outpatient clinic

Determination of sperm DNA fragmentation, as assessed by the sperm chromatin structure assay (SCSA), has become an important tool for the evaluation of semen quality. The aim of the present study was to describe the biological variation of sperm DNA fragmentation in men attending an andrology clinic and to identify clinical correlates of the biological variation of sperm DNA fragmentation. For this study, two consecutive semen samples from 100 patients attending our andrology outpatient clinic were subjected to semen analysis, performed in parallel according to WHO guidelines and by SCSA. A good agreement between pairs of samples was found for SCSA-derived variables, as indicated by a significantly lower median coefficient of variation (CV) of the DNA Fragmentation Index (DFI) and the high DNA stainability (HDS) compared with WHO semen parameters. In half of the men attending our andrology clinic, however, the individual biological variation of DFI and HDS, expressed as CV of two samples, exceeded 10%. Dysregulation of spermatogenesis, as seen as testicular insufficiency or varicocele, was not associated with increased variability of DFI or HDS. A backward multiple linear regression analysis, however, indicated that the biological variation of DFI may be more profound in men with characteristics of normal spermatogenesis. In conclusion, we confirm previous reports that sperm DNA fragmentation has a lower biological variability than classical semen parameters. We hypothesize that the sperm chromatin structure may be more influenced in patients with normal spermatogenesis, whereas in men with disturbed spermatogenesis, the chromatin structure may be already so impaired that the effect of unidentified factors leading to variability of sperm DNA fragmentation in time may not be as profound.     

The effect of cigarette smoking on human seminal parameters,sperm chromatin structure and condensation

Considerable debate still exists regarding the effects of cigarette smoking on male fertility. This work aimed to explore effects of cigarette smoking on semen parameters and DNA fragmentation on 95 infertile patients who were divided into infertile male nonsmokers (45) and infertile male smokers (50). Smokers were subdivided according to a number of cigarettes smoked per day into mild (≤10), moderate (11‐20) and heavy smokers (≥21). Semen analysis, sperm chromatin condensation integrity with aniline blue staining and sperm viability were compared between the study groups. A significant decrease has been shown in sperm count (p = .006), progressive motility (p = <.001), percentage of normal forms (p = <.001) and viability (p = .002) between infertile nonsmoker and infertile smokers. The percentage of abnormal sperm chromatin condensation was significantly higher in smokers compared to nonsmokers (p = <.001). A linear correlation was detected between the extent of cigarette smoking and the degree of worsening in progressive motility (p = .001), total motility (p < .001), viability (p < .001) and normal morphology (p < .001). These results indicate that cigarette smoking has detrimental effects on semen parameters. It negatively affected all conventional semen parameters in addition to sperm chromatin condensation and sperm viability. These abnormalities were also proportional to the number of cigarettes smoked per day and to the duration of smoking.     

人精子染色质损伤及其检测的研究进展

人精子染色质损伤是常见的影响男性生殖能力的重要原因之一,受到遗传、环境和生活习惯等多因素的影响,且与男性不育、复发性流产等有着密切的联系。随着人们对精子染色质结构和功能认识的加深,对精子染色质结构分析技术的不断改进和推广,以及辅助生殖技术越来越广泛的应用,精子DNA损伤逐渐被认识到是一个新的评价精子质量的重要检测指标,对男性生殖力的评估和辅助生殖技术的选择具有重要的临床意义。     

Sperm chromatin integrity in young men with no experiences of infertility and men from idiopathic infertility couples

Damage to the genetic component of spermatozoa seems to play the main role in a majority of cases where current approaches fail to reveal the specific cause of male infertility. In this study, we compared semen quality in men assigned to two defined groups: men from couples with unexplained infertility – idiopathic infertility (A) and young men with no experiences of infertility (B). All samples were examined by standard ejaculate analysis and sperm chromatin structure assay (SCSA). Sperm chromatin damage was significantly higher in men from group A than in those from group B. Similar results were obtained by comparison of men from group A (all men were normozoospermic) with normozoospermic men from group B. According to these results, we can suppose that chromatin disorders may be the causal factor of subfertility or infertility in some of these men. No evidence for a strong association between chromatin disorders and standard parameters of ejaculates was found. We failed to confirm a relationship between smoking and sperm quality in men from any of the investigated groups. SCSA is a method that facilitates the identification of infertile men who otherwise show normal semen variables.