甲醛慢性吸入暴露致小鼠精子畸形

目的研究甲醛慢性吸入染毒致雄性小鼠生殖细胞遗传毒性。方法 SPF级雄性ICR小鼠40只,随机分为4组:低剂量组、高剂量组、阴性对照组和阳性对照组,每组10只,低、高剂量组染毒剂量分别为1、10 mg/m~3,用静式吸入的方法进行染毒,每天一次,每次2 h,连续染毒20周,阳性对照组给予环磷酰胺。小鼠染毒后继续饲养5周。观察并计算精子畸形率。结果阴性对照组、低剂量组和高剂量组的睾丸脏器系数分别为(0.81±0.13)%、(0.82±0.12)%和(0.78±0.12)%,各组之间差异无统计学意义(P0.05);阴性对照组、阳性对照组、低剂量组和高剂量组的精子畸形率分别为4.1%、6.4%、11.5%和11.3%,与阴性对照组比较,各染毒组精子畸形率明显增加,差异有统计学意义(P0.01);且染毒剂量与精子畸形率之间存在剂量-反应关系(r_s=0.888,P0.01)。结论甲醛慢性吸入染毒可引起小鼠精子畸形率升高,且染毒剂量与精子畸形率之间存在剂量-反应关系。     

三氯化铝暴露致雄性小鼠生殖细胞的遗传毒性

目的研究三氯化铝(AlCl3)对雄性小鼠生殖细胞的遗传毒性。方法将20只清洁级健康昆明种雄性小鼠按体重随机分为4组,分别为低、中、高剂量AlCl3染毒组(染毒剂量分别为50、75、100mg/kg)和阴性对照组(0.9%生理盐水),每组5只。采用腹腔注射方式进行染毒,连续染毒2d,间隔1天,持续2周。观察染毒前后体重变化,并计算睾丸系数。采用彗星试验测定Olive尾矩,采用精子核荧光染色试验测定未成熟精子率。结果染毒前后高、中、低剂量AlCl3染毒组体重与阴性对照组比较,差异均无统计学意义(P>0.05)。高、中、低剂量AlCl3染毒组睾丸系数均小于阴性对照组,差异有统计学意义(P<0.05);且体重增长值和睾丸系数随着AlCl3染毒剂量的升高而减少。彗星试验中,高、中、低剂量AlCl3染毒组Olive尾矩均长于阴性对照组,差异有统计学意义(P<0.01);且Olive尾矩随着AlCl3染毒剂量的升高而增加。精子核荧光染色试验中,高、中、低剂量AlCl3染毒组未成熟精子率均高于阴性对照组,差异有统计学意义(P<0.01);且未成熟精子率随着AlCl3染毒剂量的升高而增加。结论AlCl3暴露可对雄性小鼠生殖细胞...     

丙烯酰胺对小鼠精子的毒性作用

目的研究丙烯酰胺(AA)对小鼠精子的毒性作用,为AA生殖毒性研究提供科学依据。方法将25只7～8周龄体重为25～30g的清洁级雄性昆明小鼠随机分为阴性对照组(双蒸水),AA染毒组(染毒剂量分别为20、40、60mg/kg)和阳性对照组(环磷酰胺),每组5只。除阳性对照组采用腹腔注射染毒外,其余各组均采用经口灌胃染毒,1次/d,连续5d,于首次染毒后14d处死小鼠,观察精子数、活动度与精子形态的变化。结果各剂量AA染毒组精子数均显著低于阴性对照组,差异有统计学意义(P<0.05);中、高剂量AA染毒组精子活动率显著低于阴性对照组,精子畸形率显著高于阴性对照组,差异均有统计学意义(P<0.01)。AA染毒后精子畸形类型主要表现在头部,且以无钩和无定形为主。结论AA对小鼠精子有毒性作用,且存在明显的剂量-效应关系。     

氯化锰对雄性小鼠生殖系统的损伤作用

[目的]探讨氯化锰对雄性小鼠生殖系统的损伤作用.[方法]将60只雄性昆明系小鼠随机分为3组:高剂量、低剂量染锰组和对照组各20只,每日固定时同腹腔注射染毒,连续染6d停染1d.于染毒d60颈椎脱臼处死.每只小鼠先剥出睾丸组织以及附睾组织称重测定脏器系数变化.后分别用睾丸组织、附睾组织进行脂质过氧化损伤相关指标以及精子运动参数的测定.[结果]随着染毒剂量的加大动物体重逐渐减轻,且睾丸胜器系数的降低高剂量染毒组与对照组相比差异有统计学意义(P<0.05);T-AOC在高剂量组与对照组相比差异有统计学意义(P<0.05),MDA含量在高、低剂量组与对照组相比差异均有统计学意义(P<0.05).高剂量染毒组VCL、VSL与对照组相比有明显降低,且差异有统计学意义(P<0.05).[结论]氧化锰引起雄性小鼠的睾丸组织脂质过氧化,影响精子运动能力.     

芹菜汁对小鼠精液参数及毒性作用的影响

目的:探讨芹菜汁对小鼠精液参数以及毒性作用的影响,为芹菜的进一步开发利用提供一定依据。方法:将小鼠按体重随机分为7d、14d和28d3个时间组,再将时间组按体重随机分为4组(对照组、低剂量组、中剂量组和高剂量组),每天定时给对照组0.3ml生理盐水灌胃,剂量组给0.3ml不同浓度的芹菜汁灌胃,分别于各时间段结束后处死动物。应用WLJY-9000型伟力彩色精子质量检测系统测定精子运动参数。结果:7d组、14d组中高剂量组和中剂量组精子密度与相应对照组比较差异均有统计学意义(P<0.05)。中剂量组:VCL14d组和28d组分别和7d组比较有统计学意义(P<0.01),VSL7d组和14d组比较有统计学意义(P<0.05);高剂量组:VCL、MAD、WOB7d组和14d组比较均有统计学意义(P<0.05)。结论:芹菜汁能够影响小鼠精子运动参数,降低精子密度。     

壳聚糖对镉致小鼠睾丸损伤的干预作用

目的探讨壳聚糖对镉致小鼠睾丸损伤的干预作用。方法50只健康昆明种雄性小鼠,随机分为5组,即对照组,单纯镉染毒组,壳聚糖50、150、450mg/kg 3个剂量干预组。壳聚糖干预组分别灌胃50、150、450mg/kg壳聚糖,对照组和单纯镉染毒组灌胃蒸馏水。2h后单纯镉染毒组和各剂量壳聚糖干预组腹腔注射氯化镉0.8mg/kg,对照组腹腔注射蒸馏水。连续14d。末次染毒后处死动物,剖取附睾和睾丸,分别测定精子畸形率、睾丸超氧化物歧化酶(SOD)活力和丙二醛(MDA)含量。结果小鼠染毒14d后,各处理组小鼠精子畸形率明显升高,与对照组比较,差异有统计学意义(P<0.05);睾丸SOD活力明显下降,其中单纯镉染毒组与对照组比较,差异有统计学意义(P<0.05);其余各剂量壳聚糖干预组与对照组比较,差异均无统计学意义(P>0.05)。MDA含量明显升高,与对照组比较,单纯镉染毒组、50和150mg/kg壳聚糖干预组差异有统计学意义(P<0.05)。随着壳聚糖摄入剂量的逐渐升高,小鼠精子畸形率有所降低,睾丸SOD活力有所升高,MDA含量有所下降,与单纯镉染毒组比较,差异均有统计学意义(P<0.05)。结论壳聚糖对镉致小鼠睾丸损伤有一定的干预作用。     

芹菜对小鼠精子运动参数的亚急性影响

目的探讨芹菜汁对成年雄性小鼠精子运动参数的影响。方法将40只昆明种雄性小鼠，按体重随机分为4组，阴性对照组、低剂量组、中剂量组、高剂量组。每天定时给阴性对照组0．3ml生理盐水灌胃，各剂量组以0．3ml不同浓度芹菜汁灌胃，于灌胃56d处死动物。采用伟力精子检测系统检测精子运动参数。结果高剂量组b级精子、精子活力及精子活率增加，d级精子降低，与对照组比较差异均有统计学意义（P〈0．05）；精子密度有降低的趋势，但与对照组比较，差异无统计学意义（P〉0．05）；高剂量组精子平均路径速度（VAP）、曲线速度（VCL）明显升高，与对照组比较，差异均有统计学意义（P〈0．05）；中、高剂量组精子侧摆幅度（ALH）增大，与对照组比较，差异有统计学意义（P〈0．05）；高剂量组精子鞭打频率（SCF）降低，与对照组比较，差异有统计学意义（P〈0．05）。结论本次实验浓度的芹菜汁有降低精子密度的趋势，但可增加精子“动能”和提高精子运动能力。     

低剂量氯化镉亚慢性染毒对大鼠精子运动功能的影响

[目的]应用计算机辅助精子分析(computer assisted sperm analysis,CASA)技术研究低剂量氯化镉(Cd-Cl2)亚慢性染毒对大鼠精子运动能力的影响. [方法]40只健康雄性SD大鼠分成4组,剂量分别为0(生理盐水)、0.1、0.2、0.4 mg/kg·bw,腹腔注射染毒,每周染毒5 d(次),共染毒8周,实验结束颈椎脱臼处死,用扩散法收集大鼠附睾尾精子制成精子悬液.1:9稀释液后甩CASA仪测定反映精子活率、精子运动能力及运动方式的各项参数. [结果]各剂量组的精子活率间差异无统计学意义(P>0.05).高剂量组的快速运动精子、中速运动精子、低速运动精子百分比与对照组比较,差异无统计学意义(P>0.05).高剂量组的VAP、VSL值与对照组比较均降低,差异有统计学意(P<0.05).高剂量组的BCF与对照组比较差异有统计学意义(P<0.05).LIN随染毒剂量增加而降低(r=-0.399,P<0.05).STR、ALH、ELON在各组间的差异无统计学意义(P>0.05). [结论]0.4 mg/kg·bw的氯化镉染毒可以引起大鼠精子运动功能的下降.     

漂白剂亚硫酸钠对小鼠生殖毒性的影响

[目的]研究亚硫酸钠(Na2SO3)对小鼠精子畸形及胚胎生长发育的影响。[方法]①取25只雄性昆明种小鼠随机分成阴性对照组、阳性对照组和3个剂量组,每组5只。分别按高、中、低剂量(130,65,32．5mg/kg)Na2SO3灌胃,阴性对照组用等量生理盐水灌胃,阳性对照组用20mg／kg的环磷酰胺腹腔注射。每天灌胃(注射)4次,连续5d,第35天处死动物,取两侧附睾液涂片,观察小鼠精子畸形。②将成年昆明种小鼠按雌雄2:1同笼交配。检出受精鼠,随机分成4组,在孕期第6天分别经口灌服高、中、低剂量Na2SO3(162．5,40．63,10．16mg/kg),连续9d,每天1次;阴性对照组用等量生理盐水;每3天称重一次,记录孕鼠体重变化。③在分娩前1天取出胎鼠,记录其体重、活胎数、吸收胎数、身长、尾长、胎鼠外观畸形和内脏畸形等指标。[结果]与对照组相比,各剂量组小鼠精子畸形率明增加(P<0．05),并有剂量-反应关系;孕鼠体重和胎鼠身长、体重均明显减少(P<0．05)。[结论]亚硫酸钠对小鼠精子畸形、胎鼠身长和体重具有明显影响,表明亚硫酸钠具有一定生殖毒性。     

β-胡萝卜素遗传毒性研究

目的探讨β-胡萝卜素的遗传毒性,为其安全使用提供科学依据。方法 Ames试验,采用平板掺入法,分加和不加代谢激活系统S9 2组平行试验,受试物设5个剂量组,计数各组回变菌落数;骨髓嗜多染红细胞微核试验,受试物设3个剂量组,检测骨髓嗜多染红细胞微核率;小鼠精子畸形试验,受试物设3个剂量组,观察不同剂量的β-胡萝卜素致小鼠精子畸形的数目。上述试验均设阴性对照组和阳性对照组。Ames试验另设一组空白对照组。结果 Ames试验,β-胡萝卜素各剂量组引起的回变菌落数未超过空白对照组自发回变菌落数的1倍以上;骨髓嗜多染红细胞微核试验,受试物各剂量组微核率与阴性对照组之间差异无统计学意义(P0.05),而阳性对照组与阴性对照组之间差异有统计学意义(P0.01);小鼠精子畸形试验,受试物各剂量组小鼠精子畸形率与阴性对照组之间差异无统计学意义(P0.05),而阳性对照组与阴性对照组之间差异有统计学意义(P0.01)。结论β-胡萝卜素对所试菌株、小鼠体细胞及生殖细胞无诱变性。     

计算机辅助精液质量分析系统参数调校方法

目的：调校精子质量分析系统的体积参数和速度参数。方法：按WHO规定的标准和方法自制精液分析的标准品，分别进行人3-．分析、CASA分析精子的浓度和活力，以人工分析结果来调校精子质量分析系统的体积、速度参数。结果：自制标准品的人工分析精子浓度（×106／mL）、总活力（％）、前向运动精子（％）分别为47．6±3．3、56．6±4．9、36．4±3．8：精子质量分析系统调校前、后的精子浓度（×106／mL）、总活力（％）、前向运动精子（％）分31]为74．8±7．5、53．3±3．6、31．9±2．8和51．3±5．1、55．1±4．5、37．5±3．4。精子质量分析系统体积参数调校前计算出的精子浓度与人工分析结果间存在显著性差异（P〈0．01）。结论：虽然精子质量分析系统经过厂家的定标，但在临床使用前仍有必要进行体积和速度参数设置。     

辛硫磷对大鼠精子生成量和精子运动能力的影响

为探讨有机磷杀虫剂辛硫的雄性生殖毒性,运用动物实验研究了不同剂量辛硫磷径口染毒大鼠６０天对精子生成量和精子运动能力的影响。采用计算机辅助精子分析仪分析精了轨迹。结果显示：辛硫磷２４．５和７３．５ｍｇ／ｋｂＢＷ染毒组,大鼠睾丸精子生成量低于对照组。     

Effect of Sample Collection Site on Semen Parameters of Healthy Young Volunteers

The effect of sample collection site on semen parameters in ten men aged between 22 and 24 years was investigated. Sperm was collected at two sites: in a university hospital restroom for general use and in a one-person hospital room. Samples were collected from the same individual twice, with an interval of two weeks between collections. Semen parameters for the two sites were compared. Samples were collected after a minimum of three days and not longer than seven days of sexual abstinence. Sperm concentration did not differ significantly between the university hospital restroom location (86.8 ± 25.4 × 10⁶/ml; mean ± standard deviation) and the private hospital room (97.1 ± 72.0 × 10⁶/ml). There was no difference in the total motile sperm count or daily sperm production between the collection sites. These results suggest that the collection site has little effect on semen parameters.     

Freezing-Free Preservation of Human Spermatozoa – A Pilot Study

This study tested a method for maintaining human spermatozoa without freezing for subsequent use in intracytoplasmic sperm injection (ICSI). We demonstrated that human sperm stored in electrolyte-free solution maintain their motility and viability for at least 4 and 6 weeks, respectively. We also have shown that preserved spermatozoa are fully functional in ICSI. Sperm chromosome analysis after injection of human sperm into mouse oocytes revealed that two weeks of storage does not negatively affect sperm DNA integrity. A mouse model was used to analyze the ability of preserved sperm to participate in normal embryogenesis. Mouse sperm preserved in electrolyte-free solution in a similar manner as human sperm maintained motility for up to 3 weeks. When mouse spermatozoa stored for 1 week were injected into the oocytes by ICSI, they yielded normal blastoctysts and normal viable fetuses. The results of the study bear significance for human assisted reproduction technologies (ART) and provide clinicians and infertile patients with a new method that can simplify sperm preparation for ICSI, assisting men who are unable to provide semen on the day of assisted fertilization.     

蜱类精子的形态

作者从蜱总科的两科5属中共观察蜱精子8种,并对科、属和种之间精子的差别做了比较。硬蜱和软蜱两科精子的主要区别是:前者纤细,后者粗大,后者长度为前者的1.5～6.9倍。除血蜱属外,头尾分明是两科精子的共同特征。在硬蜱科,不同属的精子,其长度不同,其中以血蜱属的长角蜱为最长,以硬蜱属的全沟硬蜱为最短。同属不同种的精子,其类型相同,但长度各异。     

三种不同来源精子行卵胞浆内单精子注射临床结局的比较

目的：比较三种方法获得的精子行卵胞浆内单精子注射的临床结局。方法：回顾性分析笔者所在医院2011年1月-2012年12月374个卵胞浆内单精子显微注射（ICSI）治疗周期的妊娠结局,依据精子来源不同分为三组：射出精液组（A组）,经皮附睾穿刺和经皮睾丸穿刺取精子组（B组）,供精组（C组）,比较三组的受精率、卵裂率、优质胚胎率和临床妊娠率。结果：A组的优胚率高于C组（P〈0.017）,三组患者的受精率、卵裂率、临床妊娠率比较差异无统计学意义（P〉0.05）。结论：精液、附睾或睾丸精子、供精行ICSI助孕的结局相近。精子的成熟度、精子的冷冻可能并不影响最终的妊娠率。     

Analysis of Sperm Karyotypes in a Patient Treated with Griseofulvin

Griseofulvin is known to interfere with chromosome segregation by binding to microtubule-associated proteins. Studies in mouse germ cells have demonstrated that griseofulvin can induce aneuploidy (numerical chromosome abnormalities) at therapeutic concentrations. The aim of this study was to determine if chronic griseofulvin treatment led to an increased frequency of sperm chromosome abnormalities in one male subject. We analyzed 290 full sperm karyotypes using the human sperm-hamster oocyte fusion system. The frequency of X- and Y-bearing sperm was equal. There was no increase in the frequency of numerical (1.7%) or structural (9.3%) abnormalities in the subject compared to unexposed controls. Although reassuring, this is the first report on this subject and future studies are needed to assess the risk of griseofulvin.     

精子运动能力评价方法研究进展

精子运动能力评价对男性不孕症临床检验与治疗、试管婴儿临床应用、濒危动物的生殖研究有重要意义。伴随电子技术与光学技术发展,评价方法经历了最初的人眼显微镜观察方法、光散射测量到精子动态特性计算机自动分析方法,操作越来越简便、评价越来越准确和客观。本文对上述评价方法进行简单介绍。     

Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model

Abstract

The effect of different sperm washing-selection methods on sperm morphometric characteristics as a study to detect differences in the subpopulational structure has been carried out in detail in a bovine model. Cryopreserved sperm samples from 5 bulls were thawed, pooled, and processed by TALP-washing centrifugation method (TWCM), selective Percoll discontinuous density-gradient centrifugation method (PDGM), and self-migration swim-up separation method (SUMM). Live-dead assay (SYBR-14/ethidium homodimer-1), chlortetracycline assay (CTC), and sperm motility were assessed, and aliquots of sperm were processed for automated sperm morphometry analysis (ASMA) simultaneously before (raw thawed sperm used as control, RTS) and after different sperm washing-selection techniques. Deleterious effects of different methods were evident, particularly on sperm membrane integrity (p?<?0.05) and capacitation status (p?<?0.05). Moreover, each cell was measured for four primary dimensional parameters, and three shape parameters. All sperm morphometric parameters evaluated were analyzed by principal component analysis (PCA) and multivariate clustering analyses. PCA revealed two principal components for each sperm washing or separation method explaining more than the 91% of the variance. The number of subpopulations found was the same for all methods (four) except for PDGM (three). However, irrespective of the number of subpopulations defined by PCA and clustering analyses, the sperm subpopulational structure was found to be different and strongly influenced by the sperm selection procedure due to statistical differences found regarding the sperm biophysical changes induced by each method used (p?<?0.001). It is concluded that different sperm washing-selection methods commonly used during IVF process, may lead to alterations in sperm morphometric characteristics, which might explain the different results seen after IVF, since an important influence of these methods on sperm subpopulational structure has been demonstrated.     

The importance of transmission electron microscopy analysis of spermatozoa: Diagnostic applications and basic research

This review is aimed at discussing the role of ultrastructural studies on human spermatozoa and evaluating transmission electron microscopy as a diagnostic tool that can complete andrology protocols. It is clear that morphological sperm defects may explain decreased fertilizing potential and acquire particular value in the field of male infertility. Electron microscopy is the best method to identify systematic or monomorphic and non-systematic or polymorphic sperm defects. The systematic defects are characterized by a particular anomaly that affects the vast majority of spermatozoa in a semen sample, whereas a heterogeneous combination of head and tail defects found in variable percentages are typically non-systematic or polymorphic sperm defects. A correct diagnosis of these specific sperm alterations is important for choosing the male infertility’s therapy and for deciding to turn to assisted reproduction techniques. Transmission electron microscopy (TEM) also represents a valuable method to explore the in vitro effects of different compounds (for example drugs with potential spermicidal activity) on the morphology of human spermatozoa. Finally, TEM used in combination with immunohistochemical techniques, integrates structural and functional aspects that provide a wide horizon in the understanding of sperm physiology and pathology.Abbreviations: transmission electron microscopy: TEM; World Health Organization: WHO; light microscopy: LM; motile sperm organelle morphology examination: MSOME; intracytoplasmic morphologically selected sperm injection: IMSI; intracytoplasmic sperm injection: ICSI; dysplasia of fibrous sheath: DFS; primary ciliary dyskinesia: PCD; outer dense fibers: ODF; assisted reproduction technologies: ART; scanning electron microscopy: SEM; polyvinylpirrolidone: PVP; tert-butylhydroperoxide: TBHP