不育相关抗精子抗体的免疫印迹分析

目的；分析与不育相关的抗精子抗体。方法：用精子膜抗的Ｗｅｓｔｅｒｎ ｂｌｏｔ法，对７８例生育、１９８例不孕、２２例流产及４４ 晚期妊娠孕妇血清抗精子抗体进行研究，分析与不育相关的ＡｓＡｂ。结果：抗分子量８５０００、５１０００、３００００精子抗胡的ＡｓＡｂ与不孕相关；抗分子量８５０００、７９０００、６９０００、５１０００精子抗原的ＡｓＡｂ与女性不孕相关；不孕女性ＡｓＡｂ种类明显多于生育女性，抗分子     

不育不孕症生殖道衣原体及支原体的感染

目的：了解不育不孕症生殖道衣原体及支原体的感染及其与年龄、性别、原发性、继发性、不洁性接触史的关系;解脲支原体（ＵＵ）感染、生殖道炎症与精子活动率、抗精子抗体（ＡｓＡｂ）的关系,进一步探讨不育不孕的致病因素,提高诊断准确率和治愈率。方法：选用衣原体检测及支原体培养技术、抗精子抗体（ＡｓＡｂ）检测技术、明胶凝集试验（Ｇｅｌａｔｉｎｇ Ａｇｇｌｕｔｉｎａｔｉｏｎ Ｔｅｓｔ,ＧＡＴ）,试管玻片凝集试验（     

抗精子抗体对阴道黏膜的刺激性研究

研究抗精子抗体的体外杀精效果及其对金黄地鼠阴道粘膜的刺激性,以初步评价该药作为外用避孕药应用的可能性.将抗精子抗体与新鲜精液直接混合后镜检;用导管将抗精子抗体凝胶涂布于地鼠阴道内,每天一次,两周后解剖做病理切片观察并评分.抗精子抗体的的最低杀精浓度为0.31 mg/mL;抗精子抗体凝胶对地鼠阴道粘膜刺激性评分值为5.0±0.9,在可接受的范围内.抗精子抗体具有体外杀精作用,但同时也有轻度、可接受的阴道刺激性,作为外用避孕药是可行的.     

棉酚——值得开发的避孕药

棉酚具有抗生育作用。这是我国科学家首次发现的。上世纪70年代山东中草药避孕研究协作组对山东产棉区的集中不育现象进行了调查和体检。证实棉籽食品对男性生育影响较大．男性棉籽食用者．精液中无精子的占80．5％．精子数及精子活力低于正常者占16．5％．精液正常者仅占3％。     

不明原因不育症的病因学研究

应用定时曝光显微摄影法(TEP)对32例有生育力的正常男性与50例不明原因不育症患者进行精子运动特征的客观评价与分析,并同时进行精子形态学检查、精子膜 WGA 受体值测定和血清、精浆、精子膜抗精子抗体的免疫学检测。病因学研究表明,单纯由免疫因素所致不育的患者所占比例很小;不育组在精子膜 WGA 受体值上与正常对照组无显著差异;而在精子形态与运动功能方面有显著差异;导致某些不明原因不育症患者不育的因素可能是单一因素,也可能是二种以上对生育有影响的因素。TEP 是客观.定量评价精子运幼功能的一种较理想的方法,可检测出用常规精子活力评价方法难以查出的精子运动功能缺陷。在14项精子运动特征指标中,有6项指标在对照与不育组间存在显著差异,其中2项与精子畸形率的增加存在显著的负相关性(P<0.01)。     

不明原因不育症的病因学研究

应用定时曝光摄影法（TEP）对32例有生育力的正常男性与50例不明原因不育症患者进行精子运动特征的客观评价与分析,并同时进行精子形态学检查,精子膜WGA受体值测定和血清、精浆、精子膜抗精子抗体的免疫学检测。病因学研究表明,单纯由免疫因素所致不育的患者所占比例很缩小;不育组在精子膜WGA受体值上与正常对照组无显著差异;而在精子形态与运动功能方面有显著差异;导致某些不明原因不育症患者的不育因素可能是单一因素,也可能是二种以上对生育有影响的因素。应用TEP进行精子活力评价,可获得许多精子运动特征方面的信息。这种…     

重组IL-5质粒pVAX1-mIL-5增强卵透明带DNA避孕疫苗黏膜免疫效能

目的:探讨利用白细胞介素5作为DNA疫苗佐剂增强黏膜免疫反应,提高卵透明带DNA避孕疫苗抗生育效能的可行性。方法:用pVAX1-m IL-5和卵透明带避孕疫苗pVAX1-pZP3α壳聚糖纳米微粒口服免疫雌性小鼠,共设3组,每组6只。A组单独用50μg pVAX1-pZP3α免疫;B组用疫苗加等量的pVAX1-m IL-5质粒为佐剂共免疫;C组为空白对照组。每组动物接受3次免疫(第0、3、6周)。采用ELISA法检测小鼠血清及阴道冲洗液中抗pZP3 IgA抗体。第10周雌雄交配,统计雌鼠怀孕情况;第16周取双侧卵巢作石蜡连续切片,光镜观察卵巢病理学变化。结果:B组全部小鼠血清抗体呈阳性反应,A组只有3只小鼠血清抗体呈阳性反应,且B组小鼠的抗体滴度水平均高于A组小鼠;抗生育率B组小鼠为100%,A组小鼠为50%,C组小鼠为0%,2检验分析结果表明各组间的差异有统计学意义,抗生育效能与交配时抗pZP3 IgA抗体的强度有关;卵巢切片的组织病理学观察结果表明,有抗生育效能的小鼠卵巢组织可观察到各级形态正常的卵泡,无淋巴细胞浸润,与对照组及无抗生育效能的小鼠卵巢结构无差异。结论:利用重组质粒pVAX1-m IL-5与卵...     

HBV包膜大蛋白、中蛋白和HBsAg疫苗免疫原性的比较

目的:检测HBV包膜大蛋白(L蛋白)、中蛋白(M蛋白)和HBsAg疫苗的免疫原性.方法:采用本实验室表达纯化得到的HBV包膜大蛋白、中蛋白与市售疫苗(主要成分为HBsAg),以2、10g剂量免疫家兔.ELISA检测抗HBsAg抗体滴度水平变化.结果:注射了L蛋白、M蛋白和HBsAg疫苗的家兔实验组产生的抗HBsAg抗体滴度水平有着非常明显的差别.注射了L蛋白、M蛋白的家兔在14d即可检测出抗HBsAg抗体,而注射HBsAg的家兔分别在21d和28d才能检测到抗HBsAg抗体.同时注射L蛋白实验组产生的抗HBsAg抗体滴度水平要比注射同剂量HBsAg实验组高出50%,并且注射2gL蛋白的实验组比注射10gHBsAg蛋白的实验组更快出现了抗HBsAg抗体,且抗体滴度水平更高.结论:L蛋白比M蛋白及市售疫苗的免疫原性强,不但产生抗体时间提前,而且滴度高,是新一代高效乙肝疫苗的优选蛋白.     

HCV感染与多种自身抗体高检出率的关系

为探讨丙型肝炎病毒（ＨＣＶ）感染后体内出现的自身抗体的病理意义及其引起的自身免疫的发生机制。方法：对１６１例ＨＣＶ感染者进行了八种自身抗体（抗核抗体、类风湿因子、抗甲状腺球蛋白抗体、抗甲状腺微粒体抗体、抗双链ＤＮＡ抗体、抗ＲＮＰ抗体、抗Ｓｍ抗体、抗精子抗体）的检测。结果：有５２例检出６９项次自身抗体,自身抗体检出率为３２．３％,显著高于健康人对照组（Ｐ＜０．００５）（未计抗精子抗体）。结论：ＨＣＶ感染可能是诱发自身免疫反应的一个重要因素     

盐酸川芎嗪对雄性小鼠精子数量、活动率及其恢复的影响

借鉴文章实验证明川芎嗪对于小鼠的早期妊娠有一定的影响,对雄性小鼠也具有一定的生殖毒性,较长时间用药可以杀死精子,造成不育(与健康雌性小鼠自由交配不生育),但不影响雄性激素分泌,作为雄性避孕药使用有一定的优势,因此,本实验设计短期用药对小鼠精子活动率的影响及停药后的恢复情况,以观察川芎嗪对雄性的可能避孕作用.     

Fully effective contraception in male and female guinea pigs immunized with the sperm protein PH-20

Immunization of male and female animals with extracts of whole sperm cells is known to cause infertility. Also, men and women who spontaneously produce antisperm antibodies are infertile but otherwise healthy. Although the critical sperm antigens are unknown, these observations have led to the proposal that sperm proteins might be useful in the development of a contraceptive vaccine. The guinea pig sperm surface protein PH-20 is essential in sperm adhesion to the extracellular coat (zona pellucida) of the egg, a necessary initial step in fertilization. Here, we report that 100% effective contraception was obtained in male and female guinea pigs immunized with PH-20. Antisera from immunized females had high titres, specifically recognized PH-20 in sperm extracts, and blocked sperm adhesion to the egg zona pellucida in vitro. The contraceptive effect was long-lasting and reversible: immunized females, mated at intervals of six to fifteen months after immunization, progressively regained fertility.     

Abnormal expression of centrosome protein (centrin) in spermatozoa of male human infertility

To study the relations between male infertility and centrosome protein (centrin) and the functions of centrin in spermatogenesis, the matured spermatozoa of 10 normal male people and 18 male infertility patients were stained by immunofluorescence labeling antibody against centrin. The results showed that two fluorescence signal dots appeared in the normal male spermatozoa and were located at the base of flagellum. They are proximal centriole and distal centriole. However, in some spermatozoa of the male infertility, centrin protein was located abnormally at the base of flagellum and its staining signals were spread, the normal proximal and distal centrioles were confused and could not be recognized separately. These results suggest that abnormality of centrosome protein may be related to male infertility. This discovery may be used as a marker of abnormal sperm and male infertility.     

RNA结合蛋白与人类男性不育

本文概括了控制精子正常产生的诸多候选基因在染色体上的定位、演化及其编码的 RNA 结合蛋白与人类男性精子发生之间关系研究方面的最新进展,为人类男性不育症的治疗和计划生育的顺利推行奠定了坚实的理论基础并展示出光明的前景。     

Germ cell transplantation in infertility mouse

This work investigated the spermatogenesis in an infertility BALB/c-nu mouse model by reinfusing germline stem cells into seminiferous tubules. Donor germ cells were isolated from male FVB/NJ-GFP trensgenic mice. Seminiferous tubule microinjection was applied to achieve intratubular germ cell transfer. The germ cells were injected into exposed testes of the infertility mice. We used green fluorescence and DNA analysis of donor cells from GFP transgenic mice as genetic marker. The natural mating and Southern blot methods were applied to analyze the effect of sperm cell transplantation and the sperm function after seminiferous tubule microinjection. The spermatogenesis was morphologically observed from the seminiferous tubules in 41/60 （68.33%） of the injected recipient mice using allogeneic donor cells. In the colonized testes, matured spermatozoa were seen in the lumen of the seminiferous tubules. In this research, BALB/c-nu infertility mouse model, the recipient animal, was used to avoid immunological rejection of donor cells, and germ cell transplantation was applied to overcome infertility caused by busulfan treatment. These results demonstrate that this technique of germ cell transplantation is of great use. Germ cell transplantation could be potentially valuable to oncological patients.     

540例不孕症病因分析

目的:探讨不孕症的原因。方法:通过对540例不孕症患者不孕因素的回顾性分析。结果:排卵功能障碍、输卵管因素、男性因素、子宫内膜异位症为引起不孕症的主要因素。结论:不孕症的病因分类复杂,明确诊断针对病因治疗可以提高妊振率。     

抗心磷脂抗体与不明原因复发性自然流产的相关性研究

目的 探讨抗心磷脂抗体在不同孕育状态下妇女人群中的表达情况,研究抗心磷脂抗体与复发性自然流产的相关性.方法 应用前瞻对照的研究方法,收集正常生育组、早初孕组、复发性自然流产组、继发不孕组、偶发流产组5组病例.采用ELISA法检测各组病例中的抗心磷脂抗体IgG、IgM、IgA.运用统计软件,以x2检验分析各组抗心磷脂抗体IgG、IgM、IgA阳性率的差别.结果 得到抗心磷脂抗体IgG、IgM、IgA的阳性率,正常生育组：0.00％、3.70％、3.70％;早初孕组：0.94％、4.72％、0.94％;复发性自然流产组：9.76％、7.32％、2.44％;继发不孕组：3.57％、1.79％、3.57％;偶发流产组：0.00％、3.70％、7.41％.RSA组与早孕组均有统计学意义,p＜0.05;余组间比较无统计学意义.结论 抗心磷脂抗体阳性率在RSA最高19.52％、早初孕组6.60％、继发不孕组8.93％、偶发流产组11.11％、正常生育组7.40％.不明原因复发性自然流产的发生和抗心磷脂抗体的产生密切相关.     

Antibodies induced by multi?epitope vaccine showed inhibitory activity against heterologous influenza A virus (H3N2)

In this study, recognition of 4 recombinant viral proteins (GST?NHA1) by the antibodies induced by multi?epitope vaccine was testified. Inhibitory activities of these antibodies were also investigated in vitro against four heterologous influenza A viruses (H3N2). Three epitope?specific antibodies purified by affinity chromatography could reduce the plaque formation. Interestingly, the three neutralizing antibodies in combination showed obvious enhancement of inhibitory activity, suggesting that the development of recombinant multi?epitope vaccine might be an effective way against viral mutation.     

Prevention of HIV-2 and SIVsm infection by passive immunization in cynomolgus monkeys.

Infection of macaques with simian immunodeficiency virus (SIV) and human immunodeficiency virus type 2 (HIV-2) are useful models for studies of immunotherapy and vaccination against HIV as well as for testing of antiviral drugs. Vaccine research showing protective immunity in immunized monkeys has indicated that it will be possible to develop a vaccine for prevention of human HIV infection, although many hurdles remain. The design of an HIV vaccine would be helped if the basis of the protective immunity could be elucidated. Passive immune prophylaxis offers a means to determine the relative role of antibodies in protection against infection. We have studied whether a transfer of antibodies can prevent HIV-2 and SIVsm (SIV of sooty mangabey origin) infection in cynomolgus monkeys. Sera with high antibody titres were collected, heat-treated and injected into naive animals 6 h before challenge with 10-100 monkey-infectious doses of live homologous virus. All control animals treated with normal monkey serum (n = 6) or no serum (n = 39) became infected by the challenge virus, whereas five out of seven animals pretreated with antibody-containing serum at a dose of 9 ml kg-1 resisted infection. Thus passively transferred antibodies can protect against a low-dose lentivirus challenge in a nonhuman primate.     

云南大理白族和汉族弱精子症患者精子mtDNA 4977 bp缺失相关性研究

探讨云南大理白族和汉族弱精子症患者精子线粒体DNA(mtDNA) 4977 bp缺失的相关性。收集云南大理白族弱精子症患者137例,汉族弱精子症患者121例,正常对照组大理白族和白族精子活力正常人各120例,提取精液基因组DNA,采用聚合酶链式反应(PCR)技术进行精子mtDNA4977bp缺失的研究。结果显示,120例正常对照的大理白族精子活力正常人中有10例(8.33%)存在mtDNA4977 bp缺失;而137例大理白族弱精子症患者中有89例(64.96%)存在mtDNA 4977 bp缺失。两组4977 bp缺失频率比较有极显著性差异(P 0.01)。120例正常对照的汉族精子活力正常人中有7例(5.83%)存在mtDNA 4977 bp缺失;121例大理白族弱精子症患者中有85例(70.25%)存在mtDNA 4977 bp缺失。两组4977 bp缺失频率比较差异有极显著性(P 0.01)。同时发现大理白族弱精子症患者与汉族弱精子症患者mtDNA 4977 bp缺失频率无显著性差异(P 0.05)。大理白族和汉族人群mt DNA 4977 bp缺失与弱精子症的发病有明显的关联,提示mtDNA 4977 bp缺失在弱精子症的发生中可能起重要作用。     

Sperm chromatin proteomics identifies evolutionarily conserved fertility factors

Male infertility is a long-standing enigma of significant medical concern. The integrity of sperm chromatin is a clinical indicator of male fertility and in vitro fertilization potential: chromosome aneuploidy and DNA decondensation or damage are correlated with reproductive failure. Identifying conserved proteins important for sperm chromatin structure and packaging can reveal universal causes of infertility. Here we combine proteomics, cytology and functional analysis in Caenorhabditis elegans to identify spermatogenic chromatin-associated proteins that are important for fertility. Our strategy employed multiple steps: purification of chromatin from comparable meiotic cell types, namely those undergoing spermatogenesis or oogenesis; proteomic analysis by multidimensional protein identification technology (MudPIT) of factors that co-purify with chromatin; prioritization of sperm proteins based on abundance; and subtraction of common proteins to eliminate general chromatin and meiotic factors. Our approach reduced 1,099 proteins co-purified with spermatogenic chromatin, currently the most extensive catalogue, to 132 proteins for functional analysis. Reduction of gene function through RNA interference coupled with protein localization studies revealed conserved spermatogenesis-specific proteins vital for DNA compaction, chromosome segregation, and fertility. Unexpected roles in spermatogenesis were also detected for factors involved in other processes. Our strategy to find fertility factors conserved from C. elegans to mammals achieved its goal: of mouse gene knockouts corresponding to nematode proteins, 37% (7/19) cause male sterility. Our list therefore provides significant opportunity to identify causes of male infertility and targets for male contraceptives.