大鼠睾丸组织uPA基因启动子区的克隆与分析

目的:克隆大鼠睾丸组织uPA基因的启动子区并对该序列进行分析。方法:从大鼠睾丸组织中提取基因组DNA,以其为模板设计uPA基因引物,运用降落PCR法扩增uPA基因5'端上游的真核转录调控序列。测序得到的PCR产物用启动子区的分析软件分析,并与其DNA序列进行比对。结果:得到的uPA基因长度为1572bp(登录号X65651)。经软件分析:该序列包含uPA基因完整的开放阅读框(ORF)、21bp的外显子部分,1551bp区域为转录起始的上游部分在其5'端UTR区的-30bp位置有一个不典型的TATA盒,其上游有非常明显的GC盒及启动子区常见的AP1(activeprotein1)、SP1等结合位点。结论:成功克隆了大鼠睾丸组织uPA基因启动子区。分析表明,该片段包含真核转录调控区域、不典型的TATA盒、典型的GC盒及启动子区所常见的AP1、SP1等结合区域。     

Urgent need for contraceptive education and services in Chinese unmarried undergraduates: A multi-campus survey

In order to ascertain prevalence rate of premarital sexual intercourse,unintended pregnancy and abortion,and evaluate associated factors of unintended pregnancy among undergraduates from all over China,the representative sample of unmarried undergraduates was obtained by using a multi-stage,stratified,probability cluster design,and data were collected by using a survey questionnaire.62 326 available responders were gained.11.6% of them acknowledged having experiences of premarital sexual intercourse(standardized prevalence rate of sexual intercourse was 13.8%).31.5% of students active in premarital sex acknowledged undergoing unintended pregnancy.76.2% of pregnant students selected abortion to end it.Of students active in premarital sex,46.2% used contraception at the first sexual intercourse,28.2% replied "always" using contraception in sexual intercourse.The rate of using condoms,oral contraceptives(OCs),and withdrawal among students who had used contraception was 52.0%,31.0%,and 27.2% respectively."No preparation for sex"(40.3%),"pleasure decrement"(32.1%),"won’t-be-pregnancy in occasional sexual intercourse"(30.2%) were their common excuses for using no contraception.The identified risk factors for unintended pregnancy among students active in premarital sex by multivariate analysis were as follows:having no steady lover [having no steady lover vs having a steady lover:odds ratio(OR),1.875;95% confidence interval(CI),1.629-2.158],unaware of the course of conception(unaware vs aware:OR,2.023;95% CI,1.811-2.260),considering abortion not endanger women’s physical and mental health(no endangerment vs endangerment:OR,2.659;95% CI,2.265-3.121),nonuse of contraception(never use vs always use:OR,1.682;95% CI,1.295-2.185).Medical students were not less likely to experience an unintended pregnancy than nonmedical students(OR,1.111;95% CI,0.906-1.287).The substantial proportion of unintended pregnancy among undergraduates indicates a need for convenient and targeted contraceptive education and services.     

金叶败毒治疗孕早期豚鼠巨细胞病毒感染的研究

目的：探讨清热解毒中药金叶败毒抗孕早期豚鼠巨细胞病毒(guinea pig cytomegalovirus，GPCMV)感染的效果及孕期用药的安全性。方法：①随机选择妊娠早期雌豚鼠75只分成3组，每组25只。空白对照组：不接种病毒。阳性对照组：每只腹腔接种GPCMV悬液1 mL(107 TCID50)。治疗组：按阳性对照组接种病毒，同时灌胃给予中药金叶败毒(tid，连用14 d)。20 d后处死豚鼠，采集血液及各脏器组织，检测感染情况。②随机选取受孕后第1～18天(小鼠器官发生期)的雌鼠90只，随机分成3组，每组30只。空白对照组不给药。阳性对照组给予致畸阳性对照药维生素A，100 mg·kg 1·d 1，每日分3次灌胃给药。治疗组根据小鼠灌胃最大容积给予金叶败毒，将0.5 g药溶于37℃温开水0.5 mL，tid，连用18 d。追踪母鼠一般情况及胎仔生长发育状况。结果：①阳性对照组和治疗组的血液GPCMV DNA阳性率分别为100.0%(25/25)和44.0%(11/25)，差异有极显著性(χ2＝3.94，P＜0.01)，阳性对照组出现被毛耸立等感染征象96.0%(24/25)，死亡4.0%(1/25)，治疗组无任何感染征象。②小鼠器官发生期给药不增加胎仔生长发育异常的风险。结论：中药金叶败毒具有抗孕早期GPCMV感染的效果，孕期用药比较安全。     

巨细胞病毒感染与男性生殖

生殖道的巨细胞病毒(cytomegalovirus,CMV)属于疱疹病毒家族成员,为线性双链DNA,在人群中普遍存在,男女感染比例一致[1].研究表明,男性生殖器官是CMV的攻击靶器官之一,精液亦为CMV感染的重要传染源;精液中的CMV不仅可通过性交活动传播至女性生殖道造成宫内感染,还能在男性生殖道的任何部位感染,对男性生殖功能造成影响[2-5].     

中药金叶败毒防治巨细胞病毒宫内感染的动物实验初探

人巨细胞病毒 (HCMV)宫内感染是导致出生缺陷的重要原因之一[1] ,但目前缺乏安全有效的治疗药物。本研究应用豚鼠巨细胞病毒 (guineapigcytomegalovirus,GPCMV)宫内感染模型 ,研究中药金叶败毒防治GPCMV宫内感染的效果。报道如下。1.细胞和病毒 :豚鼠胚肺细胞株和GPCMV标准株(ATCC 2 2 12 2 ) ,均经湖北省预防医学科学院病毒研究所培养传代 ,病毒滴度 10 6～ 10 7TCID50 / 0 1ml。2 .药物 :金叶败毒由华中科技大学同济医学院附属同济医院提供 ,制成流浸膏状 ,含生药 3g/ml。致畸阳性对照剂维生素A由南京市鱼肝油厂生产 ,含维生…     

Effects of Murine Cytomegalovirus Infection on Sperm Viability in Mice

In order to explore the effects of testicular infection of murine cytomegalovirus （MCMV） on mature sperm viability at different periods following MCMV inoculation in mice, 91 BALB/c mice without MCMV infection were randomly divided into two groups： an experimental group （n= 56） and a control group （n=35）. The mice in the experimental group were treated by inoculating MCMV intratesticularly, while those in the controlled group were directly inoculated with DMEM without MCMV. The mice in both groups were sacrificed separately on the day 1,1.5, 2, 4, 6, 9 and 14 post-inoculation （D1, 1.5, 2, 4, 6, 9 and 14 PI）. The MCMV M83 mRNA gene was detected in the testis by in situ hybridization （ISH） with MCMV late-mRNA probe labeled with digoxin. Sperm viability of mature sperm in the epididymis cauda was measured. The results demonstrated the positive signal of ISH of MCMV was found mainly in the cytoplasm of the testicular interstitial cells and spermatogenic cells in the experimental group. Compared with that in the controlled group, the sperm viability in the experimental group was decreased significantly on D1 PI and D1.5 PI （P〈 0.05）. No statistically significant difference in the sperm viability was found after D2 PI between two groups （P〉0.05）. This suggested that sperm viability in mice might be descended significantly shortly after MCMV infection and might return to normal with time, indicating that MCMV acute infection might temporarily degrade sperm quality and influence procreation transiently.     

睾丸巨细胞病毒感染对精子存活率影响的实验研究

目的:探讨小鼠睾丸感染鼠巨细胞病毒(MCMV)对附睾尾部成熟精子存活率的影响。方法:运用酶联免疫吸附试验(ELISA)筛选出无MCMV感染史的BALB/c小鼠分为2组:对照组40只,睾丸直接接种不含MCMV的细胞培养液;实验组64只,睾丸直接接种MCMV悬液;分别于接种后第1、2、4、6、9、14、21、38d处死小鼠,行睾丸组织病理学检查,同时取附睾尾部成熟精子进行精子存活率的检测。结果:实验组小鼠睾丸间质细胞中出现特异性MCMV嗜碱性包涵体,生精细胞空泡变性、排列紊乱。与对照组比较,实验组精子存活率在接种病毒后的第1d(D1PI)显著降低,存活率由71.42%下降至56.04%(P<0.05);D2PI至D38PI时两组小鼠精子的存活率比较无统计学意义(P>0.05)。结论:小鼠睾丸MCMV早期感染可造成精子存活率的显著下降,但随感染时间延长,精子存活率恢复正常;提示CMV感染可能短暂性地影响生育。     

胎儿宫内发育迟缓的病理生理变化

胎儿宫内发育迟缓 (ｉｎｔｒａｕｔｅｒｉｎｅｇｒｏｗｔｈｒｅｔａｒｄａｔｉｏｎ ,ＩＵ ＧＲ) ,是围生期主要并发症之一 ,我国的发病率平均为6 39%,其围生儿病死率为正常儿的 46倍。ＩＵＧＲ不仅近期会影响胎儿宫内的生长发育 ,而且远期还能影响儿童和青春期的体能和智能发育 ,因此 ,是高危妊娠中的重大问题 ,并越来越受到产科工作者的高度重视。导致ＩＵＧＲ是多方面的 ,与孕妇、胎盘和胎儿三方面的功能密切相关 ,但多数能在胎盘及其附属的病理变化上反映出来。1　第二次绒毛滋养细胞迁移未完成　　保证胎儿宫内生长发育的最佳环境 ,主…     

异常妊娠及自然流产次数与风疹病毒感染关系的初步探讨

目的初步探讨异常妊娠及自然流产次数与风疹病毒(RV)感染的关系。方法用ELISA法检测749例有异常妊娠史的育龄妇女和305例正常对照组妇女RV抗体RV-IgM、IgG,同时检测其中361例自然流产少于3次和52例自然流产3次以上的育龄妇女,并结合RT-PCR技术对12例IgM阳性的自然流产组织进行检测,数据行卡方统计。结果有异常妊娠史妇女与正常对照组妇女RV的原发感染率分别为2.267%、0.656%(P<0.001),复发感染率分别为8.545%、1.693%(P<0.001)。自然流产少于3次组妇女与3次以上组妇女RV的原发感染率分别为2.214%、7.692%(P<0.05);复发感染率分别为6.648%、15.38%(P<0.05);12例IgM阳性自然流产组织RT-PCR检测RV-RNA阳性为9例,感染率为75.0%(9/12)。结论异常妊娠与风疹病毒感染有相关性,自然流产次数与风疹病毒感染呈正相关。     

巨细胞病毒体外感染对人精子存活率及其运动参数与超微结构的影响

 目的 探讨人巨细胞病毒(HCMV)感染对人精子存活率、精子各运动参数及其超微结构的影响.方法 将HCMV与10例经过上游法优化的正常人精子体外共同孵育,于0、1、2、3、4 h分 别取样,检测精子存活率,并应用计算机辅助精子分析系统(CASA)测定精子直线运动速度(VSL)、曲线运动速度(VCL)、平均路径速度(VAP)、头部侧摆幅度(ALH)和前向运动百分率(a+b 级精子百分率);同时,电镜观察HCMV感染不同时间人精子超微病理结构的改变.结果 与空白、阴性对照组比较,人精子存活率在高浓度HCMV感染后1、3 h时明显降低,差异显著(P＜0.05),4 h时差异非常显著(P＜0.001).a+b级精子百分率仅在高浓度HCMV感染4 h时明显降低,差异显著(P＜0.05).精子VSL在各时间点各组比较差异均无统计学意义(P＞0.05).精子VCL在高浓度HCMV感染3 h时明显下降,差异显著(P ＜0.05),4 h时显著下降,差异非常显著(P＜0.001).而中浓度HCMV感染4 h时,精子VCL才下降,差异显著(P＜0.05).精子VAP在高、中浓度HCMV感染2、3、4 h时明显降低,差异显著(P＜0.05).精子ALH在高浓度HCMV感染3、4 h时明显下降,差异显著(P＜0. 05).电镜结果显示:人精子超微结构的异常改变在高浓度HCMV感染时常见,4 h时严重;主要表现有精子核解聚、空泡增多、顶体破裂、质膜破损、线粒体肿胀及其排列紊乱.结论 HCMV体外感染3～4 h可造成人精子存活率与运动参数及精子超微结构的异常,且这种改变与HCMV浓度存有一定的量效关系,高浓度HCMV感染的影响结果显著.