从非阻塞性无精症患者睾丸组织中分离出生殖干细胞并在体外分化成单倍体生殖细胞

非阻塞性无精症（NOA）患者能够通过胞浆内单精子注射（ICSI）或圆形精细胞卵浆内注射（ROSI）成功受孕，大多数成熟障碍（MA）或睾丸支持细胞综合征（SCOS）患者的精子在初级精母细胞阶段培养生殖细胞获得单倍体细胞而受孕。生殖干细胞（GSC）在成熟精子生成和自我更新间保持平衡，GSC自我更新生成的精子细胞移至不育男性的输精管。起源于胚胎干细胞单倍体生殖细胞在卵圆精子阶段在体外自发分化，引导生殖细胞发育、改变和生殖腺基因的改变。     

烷化剂类药物对睾丸生殖功能影响

烷化剂类药物是常用抗肿瘤药物和免疫抑制剂,通过诱导生殖细胞凋亡,干扰精原干细胞(SSCs)自我更新增殖分化功能,最终导致无精子症或少精子症.同时也损伤生殖细胞遗传物质,导致子代遗传缺陷或畸形.烷化剂对睾丸生精功能损伤存在量效和时效关系,并与睾丸生殖细胞成熟阶段有关.烷化剂对生殖细胞损伤也表现出相应的内分泌变化.了解烷化剂损伤男性生殖功能的机制、影响因素以及临床评价或监测的内分泌指标,有利于临床早期监测并干预.     

烷化剂类药物对睾丸生殖功能影响

烷化剂类药物是常用抗肿瘤药物和免疫抑制剂，通过诱导生殖细胞凋亡，干扰精原干细胞（SSCs）自我更新增殖分化功能，最终导致无精子症或少精子症。同时也损伤生殖细胞遗传物质，导致子代遗传缺陷或畸形。烷化剂对睾丸生精功能损伤存在量效和时效关系，并与睾丸生殖细胞成熟阶段有关。烷化剂对生殖细胞损伤也表现出相应的内分泌变化。了解烷化剂损伤男性生殖功能的机制、影响因素以及临床评价或监测的内分泌指标，有利于临床早期监测并干预。     

干细胞向雌性生殖细胞分化的研究进展

干细胞是一类具有自我更新和多向分化潜能的细胞。根据其来源不同,干细胞可分为胚胎干细胞、成体干细胞以及诱导性多能干细胞。生殖细胞系负责跨代传递遗传和表观遗传信息,以确保新的正常个体产生。人类辅助生殖技术(ART)虽可解决部分难治性不孕不育患者的生育问题,但尚不能解决由于卵巢早衰生殖细胞缺乏导致的不孕,如果在体外可以将干细胞定向诱导分化为生殖细胞,则可能通过ART帮助卵巢早衰患者获得健康后代。雌性配子发育经历了多个严格、复杂的过程,包括原始生殖细胞(PGC)特化、增殖、迁移到生殖嵴并最终分化为成熟的卵母细胞。然而具体过程尚不明确。近年来学者已建立了干细胞向雌性生殖细胞分化的体外模型,并取得了长足进步。     

干细胞诱导分化生殖细胞的研究进展

干细胞是一类具有自我更新和分化潜能的细胞,包括胚胎干细胞和成体干细胞。胚胎干细胞是从囊胚内细胞团中获得的具有全能性的细胞,具备了分化成机体所有细胞类型的潜能,包括生殖细胞。近年来人类胚胎干细胞的研究取得了突破性进展,能在体外培养成精子细胞及成熟的卵母细胞,但存在免疫排斥、致瘤性和伦理学等问题,使其应用在临床治疗方面受限。成体干细胞是一种多能干细胞,理论上成体干细胞在一定条件下也可分化为生殖细胞。成体干细胞则可克服这些缺点,应用前景日益被看好。若将成体干细胞向生殖细胞的分化应用于临床,使分化成的生殖细胞能发育成具有生命的子代,那将为不育提供一条新的治疗途径。综述了胚胎干细胞及成体干细胞向生殖细胞分化方面的研究进展。     

胚胎干细胞体外分化为原始生殖细胞与精子

胚胎干细胞(ESC)是全能干细胞,理论上讲可分化为全部生殖层的所有组织细胞包括生殖母细胞,进而分化出成熟精子.结合最近研究进展对胚胎干细胞体外分化为原始生殖细胞(PGC),进而分化出可使成熟卵母细胞受精的精子,以及胚胎干细胞和由胚胎干细胞体外分化的原始生殖细胞(PGC)和精子的鉴定,以及促进ESC体外分化为PGC的影响因素和ESC体外分化为精子的意义和存在的问题做一综述.     

胚胎干细胞体外分化为原始生殖细胞与精子

胚胎干细胞（ESC）是全能干细胞，理论上讲可分化为全部生殖层的所有组织细胞包括生殖母细胞．进而分化出成熟精子。结合最近研究进展对胚胎干细胞体外分化为原始生殖细胞（PGC）。进而分化出可使成熟卵母细胞受精的精子，以及胚胎干细胞和由胚胎干细胞体外分化的原始生殖细胞（PGC）和精子的鉴定，以及促进ESC体外分化为PGC的影响因素和ESC体外分化为精子的意义和存在的问题做一综述。     

干细胞向生殖细胞分化的研究进展

胚胎干细胞（ESC）是一种多能干细胞,具有向生殖细胞分化的能力,由于具有致瘤性和伦理问题,限制了ESC向临床应用发展的前景。成体干细胞（ASC）也是一类多能干细胞,理论上也能分化为生殖细胞,但诱导分化过程往往受到阻滞。诱导性多能干细胞（iPS）是一种具有多能性的重编程体细胞,一定程度上可替代ESC用于基础和临床研究。目前,全球不孕不育的发生率呈逐年递增的趋势,充分认识干细胞向生殖细胞分化的机制是解决该难题的重要理论前提。就各种干细胞向生殖细胞分化的研究进展作一综述。     

氧化应激对男性（雄性）生殖系统的影响

氧化应激可产生活性氧簇（ROS）。正常状况下，低水平ROS具有调节细胞生理功能的作用，而异常高水平的ROS对组织和细胞产生相应的危害，如信号通路异常、能量代谢失调、基因突变和蛋白质结构改变等，进而影响细胞、组织、器官乃至系统的功能。男性（雄性）生殖系统中，高水平的ROS对生殖器官和生殖细胞同样产生严重的危害，如对睾丸的危害可影响其形态学结构及类固醇激素的合成；对生殖细胞可直接损伤精子的DNA、膜脂质和蛋白的结构，使精子的畸形率大幅度上升，最终导致男性（雄性）不育。综述男性（雄性）生殖系统ROS的产生及其对生殖的影响，对了解临床男性不育症的发病机制及治疗具有重要的意义。     

精子DNA损伤的研究进展

随着男性不育症发病率的升高,精液常规检测已不足以精确评估男性生育力。近年来,精子DNA损伤检测作为一项评估精子质量及男性生育力更精准的指标,逐步成为生殖医学领域的研究热点。精子DNA损伤的机制包括精子发生异常、氧化应激损伤、精子凋亡异常等。目前,精子染色质结构分析法(SCSA)已经成为检测精子DNA完整性的金标准。精子DNA损伤可能与男性不育、辅助生殖结局及后代生长、发育相关。笔者拟就精子DNA损伤的机制、检测方法、对男性生育力的影响及其与辅助生殖技术的关系进行综述,旨在为男性不育症的临床诊断提供依据。     

Cell based vaccination using transplantation of iPSC-derived memory B cells

The recently developed induced pluripotent stem cell (iPSC) technique provides new direction for vaccination: somatic cells can be induced into iPSCs and expanded, then the cells are genetically or chemically promoted to a immune cell fate, followed with in vitro antigen presenting and processing processes to produce memory B cells that can secret functional antibodies to different pathogens; finally these cells are transplanted back to human.     

精子顶体发育相关蛋白的研究进展

受精是新生命诞生的第一步,在此过程中,精子将所携带的单倍体遗传物质与卵子的单倍体遗传物质融合成双倍体合子。哺乳动物的受精过程涉及到一系列复杂而精细的活动,如精子的超活化与获能、顶体反应以及精卵结合等。精子顶体是一种特殊的膜细胞器,具有帽状结构并覆盖在精子的细胞核前部,在受精过程中发挥着关键作用。顶体的形成主要包括囊泡形成、囊泡运输、囊泡融合和顶体与细胞核结合等过程,其独特的蛋白运输机制需要内质网、高尔基体等细胞器以及某些特殊结构之间的相互配合。顶体形成是精子细胞分化为精子的关键步骤之一,受多基因精细调控,其发育缺陷与包括圆头精子症在内的多种男性不育症有关。综述近年精子顶体发育过程中主要生物学事件的发生顺序中相关蛋白的研究,并介绍了自噬相关蛋白对顶体发育过程作用的新近研究,以期为男性不育症的诊疗提供新思路。     

Effect of temperature incubation and glycerol addition on the acrosome preservation in human spermatozoa

Cooling of mammalian sperms to 5 degrees C and subsequent heating to 30 degrees C causes alterations in the integrity of the plasma membrane producing acrosome disruption. This capacity of induction of acrosome reaction (AR) has allowed to evaluate this process in vitro, and has been correlated with induction of AR by follicular fluid and the percentage of in vitro fertilization. In this study, the time necessary to induce AR at 4 degrees C, the need of the increment in temperature to 37 degrees C, and the effect of glycerol to prevent the AR were evaluated. The sperms were selected by the Migration-Sedimentation technique, incubated in HTF and 1M glycerol at 4 degrees C and 20 degrees C for 1, 3, and 18 hours and then for 3 hours at 37 degrees C. The AR was determined by triple stain technique. Spermatozoa incubated by 18 hours at 4 degrees C increased the AR from 4.0% in the control (TO, post sperm selection) to 14.0% (p < 0.05). The subsequent incubation for 3 hours at 37 degrees C increased AR to 37.5% (p < 0.01 compared with 4 degrees C). The addition of 1M glycerol did not revert this effect. Our study confirms that the induction of AR with low temperatures requires long periods of incubation and a temperature increment to 37 degrees C. In addition, it is proposed that glycerol is not adequate for preservation of human spermatozoa at low temperature.     

High incidence of sperm dysfunction in a varicocele infertile man: case report

Studies indicate abnormal semen indicators among varicocele infertile men can be reversed to normal status after surgical repair. While semen indicators and DNA damage of sperms are reported frequently, sperm function tests are rarely performed to assess the functional status of sperms among these individuals. We report a 35-year-old male with 4 years of primary infertility who otherwise has a normal sexual life. Various analyses performed revealed the interplay of multiple abnormalities leading to the observed phenotype. The individual was diagnosed with severe sperm defects, bilateral varicocele (grade II) and endocrinopathy. The percentage of functionally normal sperms were found to be 24% for hypo-osmotic swelling, 28% for acrosome reaction and 21% for nuclear chromatin decondensation test. Cytogenetic analyses showed normal karyotype and sequence-tagged-site markers based PCR showed no deletions involving key candidate genes of the Y chromosome. A thorough investigation of infertile subjects and simple diagnostic tests are essential to detect the treatable defects, in general as well as severe infertile cases, which can improve the chances of normal conception or the success rates of in-vitro fertilization and intracytoplasmic sperm injection.     

Gender effects on the incidence of aneuploidy in mammalian germ cells

Aneuploidy occurs in 0.3% of newborns, 4% of stillbirths, and more than 35% of all human spontaneous abortions. Human gametogenesis is uniquely and gender-specific susceptible to errors in chromosome segregation. Overall, between 1% and 4% of sperm and as many as 20% of human oocytes have been estimated by molecular cytogenetic analysis to be aneuploid. Maternal age remains the paramount aetiological factor associated with human aneuploidy. The majority of extra chromosomes in trisomic offspring appears to be of maternal origin resulting from nondisjunction of homologous chromosomes during the first meiotic division. Differences in the recombination patterns between male and female meiosis may partly account for the striking gender- and chromosome-specific differences in the genesis of human aneuploidy, especially in aged oocytes. Nondisjunction of entire chromosomes during meiosis I as well as premature separation of sister chromatids or homologues prior to meiotic anaphase can contribute to aneuploidy. During meiosis, checkpoints at meiotic prophase and the spindle checkpoint at M-phase can induce meiotic arrest and/or cell death in case of disturbances in pairing/recombination or spindle attachment of chromosomes. It has been suggested that gender differences in aneuploidy may result from more permissive checkpoints in females than males. Furthermore, age-related loss of chromosome cohesion in oocytes as a cause of aneuploidy may be female-specific. Comparative data about the susceptibility of human male and female germ cells to aneuploidy-causing chemicals is lacking. Increases of aneuploidy frequency in sperm have been shown after exposure to therapeutic drugs, occupational agents and lifestyle factors. Conversely, data on oocyte aneuploidy caused by exogenous agents is limited because of the small numbers of oocytes available for analysis combined with potential maternal age effects. The vast majority of animal studies on aneuploidy induction in germ cells represent cause and effect data. Specific studies designed to evaluate possible gender differences in induction of germ cell aneuploidy have not been found. However, the comparison of rodent data available from different laboratories suggests that oocytes are more sensitive than male germ cells when exposed to chemicals that effect the meiotic spindle. Only recently, in vitro experiments, analyses of transgenic animals and knockdown of expression of meiotic genes have started to address the molecular mechanisms underlying chromosome missegregation in mammalian germ cells whereby striking differences between genders could be shown. Such information is needed to clarify the extent and the mechanisms of gender effects, including possible differential susceptibility to environmental agents.     

SperMagic在自然周期夫精宫腔内人工授精中的临床应用

目的：探讨SperMagic在自然周期夫精宫腔内人工授精中对精子活力及妊娠率的影响。方法：选择2012年3月—2012年6月进行自然周期夫精宫腔内人工授精的患者70例,随机分为2组：30例精子处理后添加SperMagic者为A组;40例精子处理后未添加SperMagic者为B组,比较2组精子活力和妊娠率差异。结果：A、B组年龄、不孕时间、体质量指数（BMI）以及精子处理前后密度、活力差异均无统计学意义（P＞0.05）;2组妊娠率差异亦无统计学意义（P＞0.05）,但A组有增高趋势。添加SperMagic后精子活力比精子处理后活力提高,差异有统计学意义（P＜0.05）。结论：在夫精宫腔内人工授精助孕技术中,精子处理后添加SperMagic可以明显改善精子的活力。     

甲醛对雄性小鼠生殖细胞的影响

为研究甲醛对雄性小鼠各阶段生殖细胞的一般毒性及遗传物质的损伤 ,探讨睾丸组织中丙二醛(MDA)、山梨醇脱氢酶 (SDH)及Cu、Zn与生殖细胞损伤的关系 ,揭示甲醛致生殖细胞损伤后的效应生物标志物及判断指标 ,给雄性昆明种小鼠甲醛染毒 ,剂量分别为 0 2 0、2 0 0和 2 0 0 0mg kg。采用腹腔注射染毒 5天 ,第 6、14日处死。用HE染色法观察小鼠睾丸组织的病理改变 ;显微镜下观察精子数及精子头畸形 ;采用SCE和微核试验 ,判断生殖细胞遗传物质的损伤 ;用MDA测试盒测定小鼠睾丸组织的MDA含量 ;用火焰原子吸收分光光度法测定生物材料中的铜、锌 ;用紫外分光光度法测定血液和睾丸组织中的SDH的活性。结果表明甲醛导致小鼠睾丸组织的病理改变以生殖细胞变性为主 ;甲醛各剂量组均引起精子数减少 ,精子头畸形率升高 ;甲醛的中、高剂量组能诱致早期精细胞微核率和精原细胞SCE频率显著升高 ;SDH的活性在甲醛三个剂量组均显著降低 ;甲醛高剂量组小鼠睾丸组织中的微量元素Cu和Zn的含量显著降低。结论提示 ,甲醛可导致雄性小鼠各阶段生殖细胞的一般毒性及遗传物质的损伤 ;SDH是甲醛致生殖细胞毒性的效应生物标志物 ;精子头畸形率是判断甲醛对生殖细胞一般毒性及遗传物质损伤的有效检测指标     

MSOME及IMSI技术在辅助生殖领域中的应用进展

精子形态与功能密切相关,精子形态异常是导致男性不育的重要原因之一。自1992年卵母细胞胞浆内单精子注射（ICSI）技术首次获得妊娠以来,许多男性因素不育夫妇通过该技术获得较为满意的妊娠结局。但在×400放大倍数下常规ICSI挑选的看似正常的精子仍可能存在超微结构的异常,这可能是导致ICSI周期失败的原因之一。活精子细胞器形态学检测（MSOME）技术是Bartoov在2001年创立的一项新技术,能在高放大倍数下更深入地观察精子细微结构,有助于挑选出形态正常的精子。精子形态选择性胞浆内单精子注射（IMSI）技术是在MSOME基础上结合传统ICSI技术发展而来,为卵母细胞ICSI提供了一个新方法。通过对MSOME和IMSI的文献进行综述,探讨其临床应用价值。     

酒精致雄性大鼠生殖细胞损伤的实验研究

目的:探讨酒精对雄性哺乳动物生殖细胞损伤的机理研究。方法:实验组用不同剂量44°白酒对成年雄性大鼠灌胃,造成饮酒后生殖细胞损伤模型,饮酒2个生精周期后断头取血并做内脏和生殖系统病理切片,光镜观察及体视学图像分析,并做交配后子鼠畸形的观察。结果:睾丸与附睾相对重量随饮酒量增大和周期延长而降低。精子活力显著低于对照组(P<0.01),精子活动率较正常对照组明显降低(P<0.05),精子畸形率明显高于对照组(P<0.01),血清LH、FSH及T均较正常水平低,病理及体视学显示高剂量组曲精小管萎缩变细,间隙增宽,心、肝、脑、肾无异常。结论:酒精对雄性哺乳动物生殖细胞有明显的损伤作用,减少饮酒对优生有重要意义。