精浆内脂质运载蛋白型前列腺素D合成酶与α-葡萄糖苷酶的相关性研究

目的分析脂质运载蛋白型前列腺素D合成酶(L-PGDS)与精浆其他参数之间的关系,探讨L-PGDS在男性生殖系统中的作用。方法分析92份精液中的精子密度、精子活力、L-PGDS浓度、酸性磷酸酶活力以及α-葡萄糖苷酶活力。依据精子密度,将标本分为3组:正常组(精子密度>20×106/ml)、寡精子组(精子密度<20×106/ml)及无精子组(精子密度为0)。彩色精子质量分析系统测定精子密度及活力,双抗体夹心酶联免疫吸附试验(ELISA)检测精浆内的L-PGDS浓度,分光光度计测定α-葡萄糖苷酶活力。结果正常组、寡精子组以及无精子组患者精浆L-PGDS浓度依次降低,差异显著(P<0.001)。L-PGDS的浓度与α-葡萄糖苷酶、精子密度及精子活力呈正相关,相关系数(r)分别为0.426、0.813和0.380。结论精浆L-PGDS浓度可作为少精子症的辅助诊断指标。     

精浆抗精子抗体阳性不育患者顶体酶、一氧化氮合酶及超氧化物歧化酶活力变化的研究

目的:研究精浆抗精子抗体(AsAb)阳性对精子顶体酶、精浆一氧化氮合酶(NOS)及超氧化物歧化酶(SOD)活力的影响。方法:精浆AsAb阳性不育者40例,对照组为40例正常生育男性。通过吸光度变化分别计算顶体酶活力(BAEE/ADH联合法)、NOS活力(氧化还原反应)、SOD活力(黄嘌呤氧化酶法)。结果:精浆As-Ab阳性组与正常生育组比较,精子顶体酶活力明显降低(P<0.01),NOS活力明显升高(P<0.01),精浆中SOD活力明显降低(P<0.01)。结论:精浆AsAb阳性引起不育可能与精子顶体酶、精浆中SOD及NOS活力改变有关。     

自身免疫性睾丸炎对精子特异性酶和生育功能的影响

目的 :研究自身免疫性睾丸炎对精子特异性酶和生育功能的影响。　方法 :复制豚鼠实验性变态反应性睾丸炎 (EAO)模型 ,采用酶动力学分光光度法和明胶固定底物薄膜法 ,观察EAO状态下精子顶体蛋白酶、透明质酸酶、精子胞质乳酸脱氢酶、附睾尾部精子和睾丸组织形态的变化。　结果 :EAO造成附睾精子顶体酶系中顶体蛋白酶、透明质酸酶和乳酸脱氢酶活性下降、附睾尾部精子质量下降、睾丸生精细胞发生退行性病变。　结论 :EAO明显影响雄性豚鼠生育力 ,睾丸生精细胞和附睾精子可能是主要作用环节。     

育精阴对精子特异性酶的影响

本实验通过复制实验性变态反应性睾丸炎（EAO）模型,采用酶动力学分光光度法以及明胶固定底物薄膜法,观察EAO状态下精子顶体蛋白酶、透明质酸酶和乳酸脱氢酶的变化,观察育精阴对精子特异性酶的影响。结果：EAO造成的附睾精子顶体酶系中顶体蛋白酶和透明质酸酶以及精子胞浆乳酸脱氢酶活性下降。在育精阴治疗后,顶体蛋白酶、透明质酸酶和乳酸胶氢酶活性显著恢复。揭示育精阴对EAO造成的免疫损伤有修复作用。     

微波辐射对雷达操作人员精子头部及尾部超微结构的影响

目的 研究微波辐射对精子超微结构的影响.方法 利用透射电镜对10例从事微波辐射工作2年以上男性新鲜精液样本中的精子头部及尾部超微结构进行观察.结果 在电镜下微波辐射后男性精子存在多种形态超微结构异常,有以下几种类型:(1)头部异常精子,包括尖头精子、头部含空泡精子.(2)顶体异常精子,包括顶体发育不良、缺失,顶体内形成包涵体精子.(3)尾部异常精子包括尾部形态异常精子(无尾精子、短尾精子)和尾部结构异常精子(线粒体缺失精子、尾部线粒体多种形态和结构异常).结论 微波辐射可导致男性精子项体、头部、尾部线粒体结构异常.     

微波辐射家兔阴囊对精子运动能力的影响

本实验用2,450MHz微波照射家兔阴囊,待睾内温度升至43(±0.1～0.2)℃时,维持0.5小时。每周人工阴道采精,以动态观察精子爬高力、精子cAMP含量及精子密度的变化。微波照射后,精子爬高力逐渐下降,至第31天降至最低水平(从照射前33.5±19.8降至12.1±17.0mm,P<0.05);精子cAMP含量明显下降(从照射前6.69±3.80降至3.61±3.28pmol/10~8精子,P<0.05);精子计数亦逐渐下降,约于照射后第4周降至最低水平(从照射前301×10~6降至63×10~6精子/ml,P<0.01)。本实验表明,微波照射后,尽管精子密度始终未降至零,但此时的精子运动机能已受损,从而会影响精子的受精能力。本文还探讨了微波照射后精子运动机能下降的可能机制。     

稀少精子体外培养液对人精子活力的研究

目的:探讨稀少精子体外培养液(以下提到处简称激活剂)对人精子活力是否有改善作用,以帮助临床医生、实验室及患者更好的选择辅助生殖方式。方法:本研究选取门诊进行精液常规检查的标本178例,其中弱精子症组151例,正常活力精子组27例。每份标本共取200μl,分成均等的两份,分别加入等体积的激活剂(实验组)和F10(1×)(对照组),然后在37℃、5%的CO_2恒温箱中共孵育30 min,观察并记录两组孵育前后精子的浓度、活力(前向运动、非前向运动及不活动精子百分比)和存活率的变化。结果:激活剂作用于弱精子症组后,精子存活率与作用前及对照组相比,无统计学差异(P>0.05);正常活力精子组呈现出相同的结果。激活剂作用于弱精子症组后,精子前向活力、非前向活力提高的幅度分别是:14.02%和4.86%,而存活型不活动精子比例降低的幅度是19.01%,差异均具有显著性意义(P<0.01);正常活力精子组结果与弱精子症组相似。不论是弱精子症组还是正常活力精子组,存活型不活动精子减少的百分比与存活型不活动精子百分比呈正相关,其r值分别为0.260(P<0.01)(相关程度较弱)和0.679(P<0.01)(相关程度较强)。结论:1激活剂不影响精子存活率;2激活剂提高了弱精子症和正常活力精子的活动力;3精液中存活型不活动精子百分比越高,激活剂作用后,其转变为前向、非前向的百分比越大;且正常活力组的这种相关性强于弱精子症组。     

微波辐射对精子运动参数的影响

微波辐射的脉冲波使用频率较广,主要用于雷达导航和通讯等。这些人员接触微波辐射后对人精子的影响已有相关报道。为了更详尽地了解精子运动异常的相关参数指标,我院生殖医学实验室自2000年1月至2002年9月间,借助计算机辅助精液分析技术(CASA)从精子运动活力和运动方式两个方面分别对随机抽取的90例非雷达作业人员和154例雷达作业人员进行精液检测及精子运动相关参数分析,以获取更有效的精子运动动力学结果。报告如下。     

巴戟天根两种提取物对微波损伤大鼠生精功能的影响

目的:探讨巴戟天根不同浓度提取物对微波损伤的雄性SD大鼠生精功能的影响。方法:40只健康雄性SD大鼠先分为正常对照组和辐射组,辐射后再将辐射组分为辐射模型组,巴戟天根水提物治疗组和巴戟天根醇提物治疗组,每组10只。辐射组应用微波信号发生器(900 HZ 1.0 W),功率密度为218μm/cm2,12 h/d,持续辐射2周。治疗组在辐射后分别给予巴戟天根水提物和巴戟天根醇提物20 g/(kg.d)持续灌胃2周。观察各组大鼠生长发育,扑捉潜伏期(CIP)和扑捉次数(CT),睾丸和附睾指数及精子形态学的差异,精子浓度、精子畸形率以及血清睾酮的浓度。结果:与正常对照组[(269.50±36.07)g]相比,辐射模型组[(254.77±20.38)g]大鼠体重稍降低,CIP延长及CT减少(P<0.05),精子浓度[(87.717±12.365)×106/ml]降低及精子畸形率[(0.126±0.100)×106/ml]明显升高(P<0.05);睾丸和附睾出现不同程度的病理损伤改变,睾丸指数降低,血清睾酮水平无明显变化。两治疗组较正常对照组体重显著下降(P<0.05),血清睾酮的水平显著升高,且与辐射模型组相比CT增加、CIP缩短、精子浓度显著升高、畸形率显著下降、血清睾酮的水平升高(P<0.05)。治疗组睾丸的病理性损伤显著修复,附睾管内除见大量精子外,还可见大量脱落的细胞。结论:巴戟天根的水提取物和醇提取物均可促进微波辐射损伤的生殖器官的修复以及精子的生成。     

微波辐射对雄性小鼠生殖系统的影响

目的观察微波辐射对雄性小鼠生殖系统的影响,并初步探讨其作用机制。方法将48只雄性昆明小鼠随机分成微波辐射低、中、高强度组(微波辐射功率密度分别为5、10、15mW/cm~2强度)和对照组,对小鼠辐射1h/d,连续30d。微波辐射结束后进行小鼠性行为能力的观察扑捉潜伏期(capture incubation period,CIP)、扑捉次数(capture times,CT)、精子相对计数、精子畸形数、超氧化物歧化酶(SOD)和丙二醛(MDA)等指标的检测。结果 15mW/cm~2强度辐射后15d小鼠的扑捉潜伏期显著延长、扑捉次数显著减少(P0.05),10mW/cm~2强度辐射后30d小鼠的扑捉潜伏期显著延长、扑捉次数显著减少(P0.05);精子相对计数明显减少(P0.05);精子畸形率明显增加(P0.05);血清和睾丸组织中SOD的活性显著降低(P0.05);血清和睾丸组织中MDA的含量显著增加(P0.05);5mW/cm~2强度辐射组上述各项指标与对照组相比均未见明显变化。结论低功率微波辐射可对雄性小鼠生殖系统产生影响,其作用机制可能与生殖细胞的氧化损伤有关。     

精子功能检测的研究——Ⅰ.底物膜技术对精子顶体酶的测定

本研究应用健康人和不育症患者的精液各30人份,用底物膜技术检测精子顶体酶,同时进行精子形态学和多次曝光摄片分析。结果表明,在精子头部周围,顶体蛋白酶和透明质酸酶活性区域呈现亮区或晕。不育症患者顶体蛋白酶反应率和反应区的平均直径显著降低(P<0.01),但是透明质酸酶无明显变化(P>0.05).不育症患者精子畸形率高于健康人.不育症患者和健康人精子运动平均速度相似,但直线运动方式的精子在前者明显减少.本文对精子的顶体酶、形态异常和运动等相互关系作了讨论.     

人精子透明质酸酶活性测定的临床意义

目的 :测定人精子透明质酸酶 (HYD)活性并分析其与精液常规参数之间的相关性。　方法 :用改良Singer法测定 14 6例男性精子HYD活性 ,常规检测精子密度、活动率和正常形态百分率。　结果 :精子HYD活性与密度、活动率及与正常形态百分率存在显著相关性 (r值分别为 0 .65、0 .63和 0 .72 ,P均 <0 .0 1)。不育男性精子密度在 4 0× 10 6 /ml以上组的HYD活性明显高于精子密度少于 2 0× 10 6 /ml以下组 (P <0 .0 1)。不育男性精子活动率 >60 %组的HYD活性明显高于 <3 0 %组 (P <0 .0 1)。　结论 :精子HYD活性测定是评价精子功能的有效指标之一     

Importance of glycosylation and disulfide bonds in hyaluronidase activity of macaque sperm surface PH-20

PH-20 is a glycoprotein located on the surface of the sperm plasma membrane and on the inner acrosomal membrane. The best understood function of sperm surface PH-20 is its hyaluronidase activity, which results in hydrolysis of the hyaluronic acid-rich cumulus matrix during sperm penetration of this extracellular oocyte investment. In this study, we investigated whether alterations in the secondary and tertiary structures of sperm surface PH-20 would affect its enzyme activity. Proteins were isolated from the sperm plasma membrane by treatment of living cells with phosphatidylinositol-specific phospholipase C (PI-PLC). PH-20 was purified from the PI-PLC released proteins by immunoaffinity chromatography. Two-dimensional electrophoresis of purified PH-20 revealed 6 isoforms with isoelectric points ranging from 5.1 to 6.0. Removal of the N-linked glycans from PH-20 with N-glycosidase F shifted the molecular weight from 64 kd to approximately 54 kd, its deduced molecular weight based on sequence analysis, suggesting that most if not all, of the potential N-glycosylation sites are linked to oligosaccharides. The lectins Con A and PSA recognized purified sperm surface PH-20 after Western blotting, suggesting that mannose is a major sugar within or at the terminal end of the linked glycan. The lectins UEA and LPA did not recognize PH-20 Western blot, suggesting that fucose and sialic acid are not terminal sugars of sperm surface PH-20. Deglycosylation of sperm surface PH-20 resulted in a complete loss of its hyaluronidase activity. The reduction of disulfide bonds with beta-mercaptoethanol or dithiothreitol also resulted in loss of enzyme activity. We conclude that the hyaluronidase activity of sperm surface PH-20 is dependent on structural features established by sulfhydryl linkages, as well as glycosylation.     

肝区微波照射所致肝脏损害的实验观察

目的 观察家兔肝区用不同时间与功率组合的2450 MHz微波照射后肝脏的损伤，为应用微波作肝脏缺血再灌注损伤保护的预处理摸索适宜条件。方法 参照临床理疗常用时间与功率，用2450 MHz微波的不同功率与照射时间对家兔肝区进行照射，24h后采血查肝功能变化并开腹肉眼观察肝脏改变。结果 采用临床常用的照射时间与功率对家兔肝区进行微波照射可造成肉眼可见的肝脏坏死。当功率为20w，照射时间为20min时．肝脏无明显肉眼损害．但血中ALT，AST及LDH有一定程度的升高。结论 功率为20W的2450MHz微波照射肝区20min无肉眼可见肝脏损害．但有一定程度的肝功能损害。     

不育男性精浆碳酸氢钠含量与精子活力等参数的关系

本文对80例不育症患者测定精浆N_aHCO_3含量,同时与粘液质量相比较。认为,精浆中N_aHCO_3含量与精子活力有着密切的关系,而与精液的PH值、粘稠度、精子密度无关,并就精浆N_aHCO_3含量测定在男子不育症诊治方面的意义进行探讨。     

The role of sperm-bound hyaluronidase in the dispersal of the cumulus oophorus surrounding rat ova

Only a very small portion (4-7%) of the hyaluronidase of rat sperm obtained from the caput or cauda epididymis was related during incubation in capacitation medium for up to 24 h at 37 degree C. A portion of the cell-associated hyaluronidase was accessible to external substrates, allowing sperm suspensions to lyse glycosaminoglycans of cumulus clots rapidly, even without capacitation. This process appears to represent the employment of an enzyme bound to the sperm cell, analogous to the use of solid-phase enzymes in current enzyme technology.     

Profile of hyaluronidase activity distinguishes carbon dioxide laser from scalpel wound healing.

Hyaluronic acid plays a key role in the process of wound repair. Deposition of this glycosaminoglycan polymer is in turn controlled by levels of the enzyme hyaluronidase. Hyaluronidase activity was examined in a rat incisional skin wound model comparing laser and scalpel wounds. A polyacrylamide gel electrophoresis (PAGE) hyaluronic acid substrate assay was used to detect differences in the rates of appearance, and level, of hyaluronidase activity in wound homogenates. The hyaluronidase activity in laser wounds appeared earlier, had a bimodal distribution, and increased to a higher level than that in scalpel wounds. The origin of hyaluronidase is not clear, but control of its appearance and modulation of its activity may be a more complex process than previously assumed.     

DNA damage in bovine sperm does not block fertilization and early embryonic development but induces apoptosis after the first cleavages

The main goal of this study was to investigate whether and at what level damage of paternal DNA influences fertilization of oocytes and early embryonic development. We hypothesized that posttesticular sperm DNA damage will only marginally affect sperm physiology due to the lack of gene expression, but that it will affect embryo development at the stage that embryo genome (including the paternal damaged DNA) expression is initiated. To test this, we artificially induced sperm DNA damage by irradiation with x- or gamma rays (doses of 0-300 Gy). Remarkably, sperm cells survived the irradiation quite well and, when compared with nonirradiated cells, sperm motility and integrity of plasma membrane, acrosome, and mitochondria were not altered by this irradiation treatment. In contrast, a highly significant logarithmic relation between irradiation dose and induced DNA damage to sperm cells was found by both terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) and the acridin orange assay. Despite the DNA damage, irradiated sperm cells did not show any sign of apoptosis (nuclear fragmentation, depolarization of inner mitochondrial membranes, or phospholipid scrambling) and were normally capable of fertilizing oocytes, as there was no reduction in cleavage rates when compared with nonirradiated sperm samples up to irradiation doses of less than 10 Gy. Further embryonic development was completely blocked as the blastocyst rates at days 7 and 9 dropped from 28% (nonirradiated sperm) to less than 3% by greater than 2.5-Gy-irradiated sperm. This block in embryonic development was accompanied with the initiation of apoptosis after the second or third cleavage. Specific signs of apoptosis, such as nuclear fragmentation and aberrations in spindle formation, were observed in all embryos resulting from in vitro fertilization with irradiated sperm (irradiation doses >1.25 Gy). The results show that sperm DNA damage does not impair fertilization of the oocyte or completion of the first 2-3 cleavages, but blocks blastocyst formation by inducing apoptosis. Embryos produced by assisted reproductive techniques (ART) could have incorporated aberrant paternal DNA (frequently detected in sperm of sub/infertile males). Analogously, in the present work, we discuss the possibility of following embryo development of oocytes fertilized by ART through the blastocyst stage before embryo transfer into the uterus in order to reduce risks of reproductive failure.