精浆中雌激素浓度与精子生成的关系

目的　探讨精浆中雌激素 (E2 )浓度与精子生成的关系及临床意义。方法　选择不育门诊中 64例男性患者精子密度 <2 0× 10 6 ·ml- 1 为少精症组 ,另选择 5 2例有生育史的健康男性精子密度≥ 2 0× 10 6 ·ml- 1 正常精子密度组 ,同时分析精子活动力和活率。应用磁分离酶联免疫分析法测定精浆中的E2 的浓度。结果　正常精子密度组精浆中E2 浓度 (2 0 .10± 2 .42 pg·ml- 1 )明显高于少精症组 (5 .62± 2 .65 pg·ml- 1 ) (P <0 .0 1) ,精浆中E2 浓度与精子密度 (r=0 .3 9,P <0 .0 1)、活动力 (r=0 .42 ,P <0 .0 1)呈正相关。结论　精浆中的E2 浓度的高低影响精子密度、活动力 ,可作为评价Sertoli细胞功能的指标之一     

特发性弱精子症患者精子DNA氧化损伤

目的:探讨特发性弱精子症患者精浆的氧化应激及其精子DNA的氧化损伤。方法:选择2010年12月至2011年3月就诊于中南大学湘雅医院生殖中心门诊的不孕不育夫妇。以28名特发性弱精子症患者为实验组,选取24名精液分析正常且有生育史的男性为对照组,收集新鲜精液。应用鲁米诺化学发光法检测原始精浆活性氧(reactive oxygen species,ROS)水平;酶联免疫分析法检测精子DNA氧化产物8-羟基脱氧鸟苷(8-hydroxy-2'-deoxyguanosine,8-OHdG)浓度。结果:1) 实验组精浆ROS水平增高,与对照组比较差异有统计学意义(P<0.01);两组前向运动精子活动率与精浆ROS水平呈负相关(r=-0.72,P<0.01)。2) 实验组精子8-OHdG浓度高于对照组,差异有统计学意义(P<0.01);两组前向运动精子活动率与精子8-OHdG浓度也呈负相关(r=-0.73,P<0.01)。3) 两组患者精浆ROS水平与精子8-OHdG浓度呈正相关(r=0.77,P<0.01)。结论:特发性弱精子症患者的精子具有DNA氧化损伤,可能与氧化应激有关。过量活性氧产生可能是特发性弱精子症患者精子活力低下的原因之一。     

精浆α-糖苷酶、果糖测定及其临床意义

目的探讨精浆ɑ-糖苷酶（ɑ-Glu）、果糖（Fru）对精子的影响及临床意义。方法采集60例正常生育男性及92例男性不育症精液,精子参数分析后分离精浆,分别测定ɑ-糖苷酶、果糖。结果正常组、不育症组精浆ɑ-Glu分别为（45.2±19.8）U/ml和（31.6±17.3）U/ml,两组间差异有统计学意义（P〈0.05）,Fru分别为（3.46±1.54）mg/ml和（3.22±2.17）mg/ml,两组间差异无统计学意义（P〉0.05）。男性不育组中少精组ɑ-Glu、Fru分别为（29.4±15.2）U/ml和（4.28±1.67）mg/ml,弱精组ɑ-Glu、Fru分别为（27.5±13.8）U/ml和（1.92±1.34）mg/ml。与正常组比较差异均有统计学意义（均P〈0.01）。结论精浆ɑ-糖苷酶、果糖测定对男性不育症原因分析有重要意义。     

不同密度绿脓杆菌体外对人精子前向运动百分率的影响

为了阐明绿脓杆菌体外对人精子的运动有无影响,先制备不同密度的绿脓杆菌悬液,再将其在体外与10例健康成年生育男性的经上游优化处理精子一起孵化不同时间,采用德国Mika计算机辅助的精子分析系统测定人精于前向运动百分率。结果显示:细菌与精子的比例分别为10:10、100、1000或10000孵化0、2、4和6h后,人精子前向运动百分率无明显变化。这说明当绿脓杆菌与人精子比例≤1:1孵化6h以内时,绿脓杆菌体外对人精子前向运动百分率无明显影响。     

男性不育者精浆及精子乳酸脱氢酶活性的研究

目的探讨男性不育症患者精浆及精子中乳酸脱氢酶同工酶X（LDH-X）活性,分析乳酸脱氢酶同工酶活性与男性不育的关系。方法采用聚丙烯酰胺凝胶电泳、全谱酶联染色等方法检测精浆及精子中的LDH-X活性并计算其比值;按照WHO人类精液实验室手册要求对精液进行常规分析。结果 124例不育组精浆LDH-X活性（827.3±46.8）U/L与60例生育组LDH-X活性（616.1±22.4）U/L比较,差异有统计学意义（P〈0.01）,不育组精子LDH-X的活性（8.45±3.62）mU/106小于生育组（16.1±10.6）mU/106,差异有统计学意义（P〈0.01）。两组的精浆及精子中LDH-X的比值比较,差异有统计学意义（P〈0.01）。两组的精液密度、精子活动率及精子活力无统计学差异。结论精浆及精子LDH-X活性检测能够反应精子的质量、生育功能以及对受精过程的预期,对选择辅助生育的方法有指导意义。     

精浆锌和前列腺特异性抗原含量与精液液化及精子活力的关系

目的 探讨精浆锌(Zn2+)和前列腺特异性抗原(PSA)与精液液化及精子活力的关系.方法 筛选精液液化异常患者(异常组)和精液液化正常男性(正常组)各30例,精液常规分析后提取精浆,-20℃保存.采用原子吸收光谱法测定精浆Zn2+含量,ELISA法测定精浆PSA含量,对比分析两组测定结果.结果 异常组精浆Zn2+、PSA含量均明显低于正常组.异常组精浆Zn2+平均含量为(82.50±0.72)μg/ml,正常组为(120.43±0.52)μg/ml,两者差异有统计学意义(P<0.01).异常组精浆PSA平均含量为(0.68±0.14)mg/ml,正常组为(1.21±0.21)mg/ml,两者差异有统计学意义(P<0.05).异常组精子活力较正常组低(P<0.05).结论 精浆中Zn2+及PSA含量均偏低致精液液化异常并影响精子活力.     

不育男性精浆抗流行性腮腺炎病毒抗体测定结果分析

目的观察流行性腮腺炎病毒抗体在男性生殖道分泌液中的分布状况,探讨其对男性生殖功能的影响。方法采用微孔板酶联免疫吸附法(ELISA)对30例生育男性和110例不育症患者精浆中抗流行性腮腺炎病毒抗体免疫球蛋白G(IgG)、免疫球蛋白M(IgM)进行了检测。结果不育症患者精浆中腮腺炎病毒IgG阳性者61例(55.5%),抗体平均水平为367 U/ml;IgM阳性者9例(8.2%),抗体平均水平为56U/ml;IgG和IgM同时阳性者4例(3.6%);生育组仅6例(20.0%)IgG呈现阳性,抗体平均水平为148 U/ml,明显低于不育症者。不育症组IgG阳性率以无精子症组最高,IgM阳性率以少精子症组最高。精浆中IgG在精子活动力正常组和不良组、精浆AsAb阳性组和AsAb阴性组之间存在显著性差异(P<0.05)。IgM在WBC精液组和非WBC精液组、精浆AsAb阳性组和AsAb阴性组之间存在显著性差异(P<0.05或P<0.01)。IgG和IgM同时阳性在WBC精液组和非WBC精液组之间存在显著性差异(P<0.05)。结论精浆中腮腺炎病毒IgG阳性率与精子密度呈正相关。男性感染腮腺炎病毒所致的睾丸炎,可以引起精子数量严重减少或无精子症,并且可以影响精子的活动力,对以后生育功能可能带来不良的影响。     

Effect of seminal redox status on lipid peroxidation,apoptosis and DNA fragmentation in spermatozoa of infertile Saudi males

Objectives:To assess the effect of seminal redox status on lipid peroxidation (LPO), apoptosis and integrity of sperm DNA in infertile males.Methods:In this case-control study, the total antioxidant status (TAS) and reactive oxygen species (ROS) levels were analyzed within the seminal plasma of fertile normozoospermic, n=40 and infertile (asthenozoospermic, n=30; oligoasthenoteratozoospermic, n=30) males. Additionally, the level of 4-hydroxynonenal (4-HNE), DNA fragmentation, and caspase-3 activity were estimated in the spermatozoa.Results:Significantly (p<0.001) increased seminal ROS level with decreased TAS scores was observed in the infertile groups compared to normozoospermics. The infertile males showed marked elevated (p<0.001) levels of 4-HNE, DNA fragmentation and caspase-3 activity compared to normozoospermics, which was positively correlated to increased seminal ROS levels and negatively to the TAS score in the studied groups. Seminal ROS level was significantly inverse correlated to the semen parameters. Additionally, a strong negative correlation between DNA fragmentation, LPO, caspase-3activity and seminal parameters were observed.Conclusion:Seminal oxidative stress is a potential risk factor for LPO, DNA damage, and apoptosis in spermatozoa, which can affect semen quality and male fertility. Thus, in addition to conventional seminological parameters, measurement of seminal oxidative stress and sperm DNA integrity may also be employed to investigate the functional integrity of spermatozoa at the molecular level.     

Pharmacological Investigation of Voltage-dependent Ca^2＋ Channels in Human Ejaculatory Sperm in vitro

The types of the voltage-dependent calcium channels (VDCCs) in human ejaculatory sperm and the effects of calcium channel blocker (CCB) on human sperm motility parameters in vitro were investigated. The human sperm motility parameters in vitro in response to the pharmacological agents nifedipine (NIF, inhibitor of L-type VDCC) andω-conotoxin (GVIA, inhibitor of N-type VDCC) were compared and analyzed statistically. The results showed that NIF (1, 5, 10μmol/L) could not only significantly affect human sperm's shape but also spermatozoa motility after incubated at least 10 min in vitro (P<0.001). GVIA (0.1, 0.5 and 1μmol/L) could just only significantly affect human sperm's progressive motility (a % b %) after incubated for 20 min in vitro (P<0.01), but they both could not significantly affect spermic abnormality rate. It is suggested that L-type VDCC, non L-type VDCCs and isoform of L-type VDCC exist in the cell membrane of human sperm solely or together, and they participate in the spermic physiological processes especially the spermic motility.     

精索静脉曲张患者精子DNA碎片率、精浆丙二醛含量及精子活力的变化

目的：探讨精索静脉曲张对精子DNA碎片率（DFI）、精浆丙二醛（MDA）含量及精子活力的影响。方法：精索静脉曲张的不育患者按精索静脉曲张临床分度分为Ⅰ度、Ⅱ度和Ⅲ度组,采用染色质扩散实验（SCD）对85例精索静脉曲张所致男性不育患者和22例健康生育男性进行精子DFI检测;采用硫代巴比妥酸法（TBA）测定精浆MDA的含量;采用计算机辅助精液分析仪按照WHO标准测定精子活力。结果：和正常生育男性相比,Ⅰ度、Ⅱ度和Ⅲ度组的精子DFI均升高（P〈0．05）,但各组间差别无统计学意义（P〉0．05）;和正常生育男性相比,Ⅰ度、Ⅱ度和Ⅲ度组的精浆MDA含量均升高（P〈0．05）,各组间差别有统计学意义（P〈0．05）;和正常生育男性相比,Ⅰ度、Ⅱ度和Ⅲ度组的前向精子活动百分率均下降（P〈0．05）,各组间差异有统计学意义（P〈0．05）。结论：精索静脉曲张可导致精子DFI和精浆MDA含量升高,而前向活动精子百分率下降;随着精索静脉曲张临床分度的增加,精浆MDA含量明显升高、前向活动精子百分率明显下降,而精子DFI升高不明显。     

左卡尼汀在特发性弱精子症患者中的水平测定

目的 探讨特发性弱精了症患者中左卡尼汀水平与精子活力的关系.方法 实验组:特发性弱精子症50例,对照组:有致孕史男性10例.采用高效液相色谱法测定精浆左卡尼汀水平,并检测精液参数(精液量、精子密度、精子前向运动百分率).结果 两组精子密度、精子前向运动百分率差异有统计学意义(P＜0.01);左卡尼汀水平有显著性差异.结论 特发性弱精子症患者的左卡尼汀水平明显降低,与精了活力低密切相关,提示左卡尼汀治疗特发性弱精子症为有效的治疗方法.     

男性不育不同精液参数患者精浆α-糖苷酶活性测定

目的：测定男性不育不同精液参数患者精浆α-糖苷酶活性.方法：对480例不育男性及45例正常对照受试者的精液标本,进行精液常规分析及α-糖苷酶活性测定.比较不同精液液化程度、精子密度、活动率和活力患者α-糖苷酶活性.结果：①按液化程度分组,不育各组患者精浆α-糖苷酶活性与正常对照组差异无统计学意义（P＞0.05）;②按精子密度分组,梗阻性无精子症组精浆α-糖苷酶活性明显低于其他组（P＜0.05）;③按精子活动率或活力分组,低活动率和低活力组精浆α-糖苷酶活性明显低于正常对照组、正常活动率和正常活力组（P＜0.05）.结论：精浆α-糖苷酶活性可以作为判断男性生育力的参考指标.     

精浆弹性蛋白酶水平、白细胞定量与男性不育的关系

目的：探讨男性不育患者精浆弹性蛋白酶水平、白细胞定量与精液质量的关系。方法：对137例男性不育患者采用酶联免疫吸附试验（ELISA）进行精浆弹性蛋白酶水平检测,采用过氧化物酶染色法进行精液白细胞定量测定,采用清华同方CASAS-QH-Ⅲ计算机辅助精液分析仪进行精液质量分析。结果：精浆弹性蛋白酶水平异常组与对照组精液pH、精子密度、a级精子百分率、精子活力比较,差异有统计学意义（P〈0.05）;精浆弹性蛋白酶水平异常的隐性感染组与确证感染组精液pH、b级精子百分率、精子活力比较,差异有统计学意义（P〈0.05）;精浆弹性蛋白酶水平与白细胞定量呈正相关（rs=0.561,P〈0.01）;正常组不育患者精液白细胞含量（〈1×10^6/ml）与异常组（≥1×10^6/ml）不育患者精液pH、a级精子百分率、精子活力比较,差异有统计学意义（P〈0.05）。结论：精浆弹性蛋白酶及白细胞含量可能与精液质量及不育关系密切。     

人精浆EGF GM-CSF NPY ET-1测定的临床实验研究

目的观察生育者与不育症者精浆中表皮生长因子(EGF)、粒-巨噬细胞集落刺激因子(GM-CSF)、神经肽Y(NPY)和内皮素-1(ET-1)等多肽类物质水平及其对精子数量和功能的影响。方法应用放射免疫分析技术(RIA)对110例不同原因引起的不育男性精浆中EGF、GM-CSF、NPY、ET-1等进行综合性测定和分析,同时与精液中精子的功能指标进行相关分析。结果不育症组精浆EGF、GM-CSF、NPY、ET-1含量均明显高于生育组(P<0.01)。在不育症组中,除EGF外,随精子密度的减少,GM-CSF、NPY、ET-1浓度均依次呈不断增高的趋势;在精子活动力与活动率及毛细管穿透力测定正常组、非WBC精液组精浆EGF含量均低于不正常(或下降)组(P<0.05);精子活动力和毛细管穿透力测定正常组、非WBC精液组精浆GM-CSF含量均低于不正常(或下降)组(P<0.05或P<0.01);NPY含量在精子活动率正常组、非WBC精液组均低于不正常(或下降)组(P<0.05);ET-1含量在非WBC精液组明显低于WBC精液组(P<0.05)。结论精浆中EGF、GM-CSF、NPY、ET-1含量增高或降低反映生殖系统局部的免疫状态和感染情况,其增高产生的毒性作用可影响精子的产生和精子的正常功能。检测精浆中EGF、GM-CSF、NPY、ET-1等物质的含量可以反映男性不育症患者的状态,有利于帮助临床进行有针对性的治疗。     

苦参总生物碱及其单体在Caco-2细胞模型的吸收特征研究

目的 建立同时测定Caco-2细胞模型中苦参碱、氧化槐果碱和氧化苦参碱的HPLC方法,探讨苦参总生物碱在Caco-2细胞模型的吸收机制。方法 利用人源结肠腺癌细胞系Caco-2细胞单层模型,研究苦参碱、氧化槐果碱和氧化苦参碱由细胞绒毛膜面供给侧（AP）→基底外侧（BL）和BL→AP侧两个方向的转运过程;HPLC-UV法测定上述3个生物碱的量;计算转运参数和表观渗透系数（P_app）。结果 苦参总碱给药后,苦参碱、氧化槐果碱和氧化苦参碱由AP→BL侧的P_app分别为（1.098±0.092）×10^?5、（1.434±0.098）×10^?5、（3.87±0.64）×10^?6cm/s,由BL→AP侧的P_app分别为（1.104±0.098）×10^?5、（1.034±0.079）×10^?5、（2.75±0.33）×10^?6 cm/s,与文献报道的单体化合物给药相比,氧化槐果碱和氧化苦参碱双向转运的P_app明显增大。苦参碱、氧化槐果碱和氧化苦参碱的表观渗透率值分别为1.01、0.72、0.71。结论 苦参总生物碱中苦参碱、氧化槐果碱和氧化苦参碱仍主要以被动吸收方式进入体内,但比各单体给药吸收更好。     

中性α－糖苷酶果糖和其它精液参数的关系

目的研究中性α－糖苷酶、果糖和其它精液参数之间的联系。方法６０３例不育病人，４７例正常生育者，按ＷＨＯ（１９９２）手册中的方法检测中性。α－糖苷酶、果糖和其它精液参数。结果在精子计数和中性α－糖苷酶活性（ｒ＝０．４５）、总活性（ｒ＝０．３８）之间，在精液体积和果糖水平之间里正相关．在精子计数和果糖浓度（ｒ＝－０．３９）　、果糖总量（ｒ＝－０．４６）之间呈负相关．前向精子活力和糖苷酶、果糖之间不相关．在ｐＨ和糖苷酶或果糖之间有抛物线的关系．结论在少精子症个体，中性α－糖苷酶和精子数减少更可能反映附睾的阻塞或附睾小管损害的变化程度。在低精液量者，精液量和果糖水平的减少更可能反映了精囊分泌功能失调的变化程度。中图分类号Ｒ６９８２     

精液粘度对精液质量的影响及相关因素分析

目的：观察精液粘度增高对精液质量的影响及相关因素分析。方法：308名男性不育门诊患者,按精液粘度正常与否分为粘度增高组和正常组,进行精液检查。结果：精液粘度正常组和增高组患者的精液量、精子密度、a级精子百分比、b级精子百分比、c级精子百分比、正常形态精子百分比、精子运动轨迹各项参数之间无统计学差异,P＞0.05;粘度增高组精液pH值、液化时间、d级精子百分比分别为7.3±0.2、22.0±4.3（min）和（59.5±15.5）%,明显高于粘度正常组,分别为7.1±0.2、20.7±5.1（min）和（54.5±19.3）%,P＜0.05;粘度增高组精子存活率为（65.2±17.0）％,明显低于粘度正常组,后者为（70.7±15.2）％,P＜0.05。精液粘度增高与精液白细胞数量无关（rs=-0.005,P值=0.927）,与精浆抗精子抗体存在正相关（rs=0.184,P值=0.001）。结论：精液粘度增高可以导致精液液化时间延长,pH值增高,降低精子存活率和活动力,与精浆抗精子抗体相关,应当引起临床医生的高度重视。     

Influence of Acupuncture on Idiopathic Male Infertility in Assisted Reproductive Technology

From Oct. 1 999to Aug. 2 0 0 0 ,2 2 cases ofidiopathic male infertility treated by acupuncturehad been observed before and after intracyto-plasmic sperm injection ( ICSI) . The purpose ofthis study was to investigate the changes in semenmarkers,fertilization rate and embryo quality be-fore and after acupuncture.1　 MATERIAL AND METHODS1 .1　 Clinical DataFrom Oct. 1 999to Aug. 2 0 0 0 ,2 2 cases ofidiopathic male infertility at Reproductive MedicalCenter in Ulm,Germany were treate…     

Mechanism of lead induced effects on human spermatozoa after occupational exposure

Objectives: Occupational lead exposure caused several types of male reproductive impairments in different working populations. In the present study we examined the paint factory workers of active reproductive age and compared the data with the non-occupationally exposed desk job holders taken as control from Bangalore, India. Materials and methods: In the above perspective, sperm cell morphology, morphometery and motile activity were assessed. Routine seminal biochemistry, cell cycle phase analysis of sperm head DNA, estimation of serum reproductive hormones and metal levels in blood and semen were also taken into account. Result: Low sperm velocity, ATPase activity, gross and forward progressive motility with high stationary motile spermatozoa revealed lowering of cellular activity after lead exposure (p<0.001), which was supported by high seminal plasma fructose level (p<0.001). Lowering of seminal plasma total protein with concomitant rise in free amino acid level was prevalent as the exposure increased (p<0.001), suggesting disturbance in cellular nutritional support essential for cellular motility. Prolonged liquefaction time, reduced semen volume and viscosity as well as altered seminal plasma protein, fructose and cholesterol level among the workers indicated dysfunction of accessory sex glands viz. prostate and seminal vesicle after occupational lead exposure (p<0.001). Deterioration of sperm count, structural abnormality of spermatozoa and sperm head DNA hyploidy was also associated with high blood and semen lead levels in the paint factory workers (p<0.001) without interfering serum FSH, LH and testosterone level (non-significant at p<0.05). Conclusion: Therefore, the present study suggested that at the present exposure level lead might cross blood-testis-barrier and increased its value in semen of the occupationally exposed paint factory workers in Bangalore, India, thereby producing detrimental effects on semen quality and sperm characteristics. Key words: Blood/semen lead level, cell cycle, paint factory workers, semen biochemistry, sperm morphology.     

Role of Urokinase-type Plasminogen Activator in the Precontact Sperm-egg Communication and Fertility of Mice in vitro

Objective To explore the role of urokinase-type plasminogen activator（uPA） in precontact sperm-egg communication and fertility of mice in vitro. Methods Firstly, sperm chemotaxis （SC） induced by uPA was assayed by measuring the sperm densities in capillaries with a descending gradient or no gradient of uPA respectively. Secondly, the role of uPAR that exists in sperm plasma membrane in SC was studied by examining the change of sperm density in capillary after incubating spermatozoa with anti-uPAR antibody. Thirdly, SC induced by eggs, which had been treated with uPA, PAl-1 and anti-uPAR beforehand respectively, was assayed to study the role of uPA in PSEC. Lastly, the fertilization capability of spermatozoa treated with uPA was examined by counting the number of fertilized eggs. Results 1）The density of spermatozoa that migrated down the gradient of uPA into the capillary was significantly lower than that into the capillary containing no-gradient uPA. 2） When uPAR of spermatozoa was inhibited by anti-uPAR antibody, the density of spermatozoa that migrated into the capillary with ascending gradient of uPA decreased correspondingly. 3） The density of spermatozoa attracted by eggs, which were treated with uPA beforehand, increased significantly than that of attracted by non-treated eggs. On the contrary, the sperm density decreased correspondingly when the egg was treated with PAI-1. 4） The number of fertilized eggs increased significantly after the spermatozoa used here was treated with uPA beforehand. Conclusion uPA could induce SC of mice sperm in vitro through the uPAR on its membrane, enhance the capability of egg inducing SC, and promote spermatozoa to fertilize eggs. Thus, uPA may act as an attractant in PSEC, increase the chance encounter of spermatozoa and eggs, therefore, enhance the fertility success correspondingly. This study, in some degree, provides an evidence that uPA may be used as a new medicine and diagnostic reagent for male infertility.