淋病奈瑟菌感染动物模型研究进展

淋病奈瑟菌容易感染人,却难以感染所有常用实验动物,因此长期以来一直缺少合适的实验动物模型,严重限制了淋病奈瑟菌的研究进展。雌激素有一定的抗炎作用,利用雌激素处理小鼠,建立淋病奈瑟菌实验感染模型,有助于淋病奈瑟菌感染机制的研究、疫苗的开发和研制、以及抗淋病药物的筛选与评价。     

淋病奈瑟菌感染动物实验模型的探讨

淋病奈瑟菌是人类淋病的病原菌,人类是自然界淋病奈瑟菌的唯一宿主,人类极易被感染,可却难以感染常用的实验动物,而淋病奈瑟菌的研究必须做大量的动物实验。由于合适的实验动物的缺乏,淋病奈瑟茵的研究受到严重阻碍。雌激素有抗炎作用,能使阴道黏膜增厚并且促使上皮细胞增生。如果将雌鼠用雌激素处理,然后用其建立奈瑟淋球菌实验动物感染模型,相信对今后研究奈瑟淋球菌将起到积极的作用,这其中包括奈瑟淋球菌的感染机制,疫苗的开发以及抗菌药物的使用。本文对此作一综述。     

解脲支原体感染动物模型优化方案研究

目的探索不同造模方法的优劣性,选择最合适的解脲支原体感染动物模型方法。方法选择SPF级BALB/c雌性小鼠、Wistar大鼠,分别以小剂量UU3菌液多次感染、大剂量UU3菌液一次感染、苯甲酸雌二醇预处理为干预措施,建立解脲支原体感染动物模型,分别于首次接种后14 d、28 d、42 d、56 d取实验动物宫颈分泌物行解脲支原体培养,取生殖器做形态学分析。结果 1BALB/c小鼠与Wistar大鼠相比,BALB/c小鼠总体一般情况较差,阴道分泌物增加,外阴溃烂明显,而Wistar大鼠外阴无明显变化。2大鼠各组阴道UU定植率比较,差异均具有统计学意义(P0.05);小剂量雌激素组与小剂量非雌激素组比较,差异有统计学意义(P0.05);大剂量雌激素组与大剂量非雌激素组比较,差异有统计学意义(P0.05);小剂量雌激素组与大剂量雌激素组比较,差异有统计学意义(P0.05);小剂量非雌激素组与大剂量非雌激素组比较,差异有统计学意义(P0.05)。小鼠各组结果同大鼠。3同样干预措施,在小剂量雌激素处理下,大、小鼠比较,差异有统计学意义(P0.05);在大剂量雌激素处理下,大、小鼠比较,差异有统计学意义(P0.05),说明同等处理措施下小鼠阴道UU定植率更高。4肉眼观:小剂量雌激素组个别大鼠生殖道组织有水肿,其余各组大鼠生殖道组织柔软,活动度好;空白组小鼠生殖道组织柔软,活动度好;小剂量雌激素组小鼠生殖道组织充血明显,输卵管积水,增粗,僵硬,弹性差;大剂量雌激素组小鼠以疏松水肿为主;小剂量非雌激素组和大剂量非雌激素组仅见阴道、宫颈轻度充血水肿。5镜下观:大鼠各干预组阴道组织无明显病理损伤,部分可见疏松组织水肿。小剂量雌激素组小鼠病理损伤严重;小剂量非雌激素小鼠组、大剂量雌激素小鼠组充血水肿程度较轻,腺体轻度扩张,炎性分泌物较多。结论 BALB/c小鼠与Wistar大鼠相比,外阴损伤明显;雌激素预处理能增加实验动物解脲支原体的定植率;小剂量多次感染比大剂量一次感染更能增加UU定值率;UU在BALB/c小鼠阴道UU定植率更高;BALB/c小鼠病理损伤严重,对动物实验研究价值更大。     

172株淋病奈瑟菌药敏试验结果分析

淋病奈瑟菌是性传播疾病的主要病原之一 ,为了给临床提供合理的用药依据 ,我们从附属医院门诊病人中分离出172株淋病奈瑟菌 ,用琼脂扩散法进行药物敏感试验 ,现报告如下。1　材料与方法1.1　标本来源　收集附属医院门诊疑为淋病奈瑟菌感染的标本 473份 ,其中男性尿道分泌物 317份 ,女性阴道分泌物15 6份。1.2　材料　淋病奈瑟菌培养基 (上海市医学化学试剂厂 ) ,药敏纸片均为标准品 (浙江天和微生物试剂有限公司 )。1.3　方法　无菌采集尿道或阴道分泌物立即接种淋病奈瑟菌培养基上 ,在 5 %～ 10 %ＣＯ2 下孵育 2 4～ 48ｈ ,挑取凸起、圆形…     

复发性念珠菌性阴道炎及其他性传播疾病的检测

目的 探讨复发性外阴阴道假丝酵母菌病患者合并滴虫、加德纳菌、淋病奈瑟球菌（NG）、解脲脲原体（UU）感染情况。为临床治疗复发性念珠菌阴道炎提供依据。方法 用涂片和培养法对108例复发性外阴阴道假丝酵母菌病患者和120例妇教人员的阴道分泌物进行病原学检测及酵母菌的培养鉴定。结果 108例复发性外阴阴道假丝酵母菌病患者中合并解脲脲原体感染的35例,对照组12例（P〈0．01）,合并滴虫感染的5例,对照组0例（P〉0．05）,合并淋病奈瑟球菌的2例,对照组0例（P〉0．05）,合并加德纳菌感染的6例,对照组10例（P〉0．05）。结论 复发性外阴阴道假丝酵母菌病患者常合并UU感染,在复发性外阴阴道假丝酵母菌病患者的诊治中有必要进行UU的检测和相应的治疗。     

淋病奈瑟菌的耐药性及产β-内酰胺酶研究

目的：了解淋病奈瑟菌的耐药性及β－内酰胺酶的产生情况。方法：对门诊1998－2001年分离的602株淋病奈瑟菌进行14种抗菌药物的耐药性分析，同时检测β－内酰胺酶。结果：药敏试验结果显示：头孢噻肟（Cefotaxime,CTX)和原始霉素（Pristinomycin,PRI)的耐药率分别为4．65％和4．98％，而甲氧苄氨嘧啶（Trimethoprim,TMP)的耐率94．52％。602株淋病奈瑟菌中β－内酰胺酶淋病奈瑟菌（PPNG）占38．87％，抗四环素淋病奈瑟菌（TRNG）占16．44％。结论：头孢噻肟和原始霉素应作为本地区目前临床治疗淋病奈瑟菌感染的首选药物。     

微波处理标本用于聚合酶链反应检测淋病奈瑟氏菌

微波处理标本用于聚合酶链反应检测淋病奈瑟氏菌阮月芹韩兆东高乐俊（附属医院检验科，滨州市２５６６０３）关键词微波；聚合酶链反应；淋病奈瑟氏菌聚合酶链反应（ＰＣＲ）检测泌尿、生殖道炎患者分泌物中淋病奈瑟氏菌ＤＮＡ（ＮＧ－ＤＮＡ）是诊断淋球菌感染的一种快速...     

淋球菌外膜蛋白Porin I多抗的黏附抑制作用研究

目的:研究兔抗淋球菌外膜蛋白Porin I(P I)的多克隆抗体阻断淋病奈瑟菌对泌尿生殖道上皮的黏附作用。方法:将自行构建表达的淋球菌GST-P I融合蛋白作为抗原免疫新西兰兔,获得兔抗P I蛋白的多抗血清,纯化后得到P I-IgG抗体。建立淋球菌感染BALB/c小鼠模型,并通过观察阴道黏膜改变、分泌物涂片、冲洗液培养及阴道组织病理变化评价兔抗P I-IgG抗体对淋球菌黏附的影响。结果:经1 m g/m l兔抗P I-IgG处理后3 h再接种淋球菌的BALB/c小鼠生殖道黏膜未见红肿及脓液,分泌物涂片及阴道冲洗液未检及淋球菌,阴道组织病理检查也未见炎症细胞浸润。而低于浓度1 m g/m l及长于3 h兔抗P I-IgG处理的小鼠阴道组织病理可检及炎症细胞,但其它检查结果阴性。结论:纯化的抗淋球菌GST-P I融合蛋白的多克隆抗体,能有效抑制淋病奈瑟菌对小鼠泌尿生殖道上皮的黏附与感染,阻断作用及持续时间与抗体浓度有关。     

苯甲酸雌二醇对腹腔镜下腹膜阴道成形术后阴道上皮的影响

目的观察苯甲酸雌二醇对腹腔镜下腹膜阴道成形术后阴道上皮的影响。方法将2003年1月至2011年3月在邢台市人民医院行腹腔镜腹膜阴道成形术的40例患者随机分成实验组与对照组,每组各20例。实验组于人工成形的阴道内涂抹苯甲酸雌二醇软膏,每日1．5g,连用3个月;对照组不用任何雌激素类药物。结果术后3个月阴道长度实验组为（9．1±0．4）cm,对照组为（9．0±0．6）cm,两组比较差异无统计学意义（P〉0．05）;但实验组阴道弹性好,并发症发生率低。实验组术后3个月阴道涂片结果为：可见大量复层鳞状上皮细胞,少量炎症细胞,阴道细胞成熟指数[Vaginal exfoliate cell maturationindex,MI（％）]为（50．2±10．5）。对照组为：可见中等炎症细胞,少许鳞状上皮细胞。结论苯甲酸雌二醇应用于腹腔镜腹膜阴道成形术后可增加阴道上皮弹性,促进人工阴道鳞状上皮化,减少并发症。     

解脲支原体血清8型感染小鼠盆腔炎症性疾病模型的建立

目的：建立解脲支原体阴道感染小鼠盆腔炎症性疾病的动物模型。方法：采用苯甲酸雌二醇预处理,给予Balb/c小鼠阴道接种解脲支原体血清8型感染液制成盆腔炎症性疾病模型,接种后1周及3周检测支原体后处死,取小鼠阴道、宫颈、子宫、输卵管组织常规病理包埋切片观察及检测支原体,并采用流式细胞术对外周血T淋巴细胞亚群进行了分析。结果：造模后小鼠阴道发红,有的分泌物增多,宫颈肥大,子宫及输卵管水肿。阴拭子培养支原体均为阳性。病理切片显示生殖器官4个部位均有明显中性粒细胞浸润。流式细胞术检测显示外周血CD4＋/CD8＋比值下降,具有统计学意义。结论：雌激素预处理下,解脲支原体阴道接种小鼠可逆行性感染引起宫颈炎、子宫炎、输卵管炎,机体免疫功能下降。此方法可成为盆腔炎症性疾病的成功造模方法之一。     

"皮肤物理抗菌膜"专利技术在小鼠生殖道淋病奈瑟球菌感染中的预防作用

目的 探讨"皮肤物理抗菌膜"专利技术在小鼠淋病奈瑟球菌(简称淋球菌)感染中的预防作用,以寻求淋病预防新方法.方法 (1)模拟雄鼠向雌鼠传播淋球菌:将淋球菌WHO-L菌株接种于经过苯甲酸雌二醇预处理的雌性BALB/c小鼠阴道.实验组小鼠阴道使用"皮肤物理抗菌膜"长效抗菌材料,对照组小鼠阴道使用去离子水.接种后第2、4、5天取阴道分泌物做淋球菌培养,比较两组检出率;接种后2～10 d监测小鼠阴道分泌物涂片染色中多形核白细胞(PMN)平均数占总细胞百分数.(2)模拟雌鼠向雄鼠传播淋球菌:用玻璃棒代替小鼠阴茎,与已感染淋球菌的雌鼠模拟性交.实验组玻璃棒用长效抗菌材料浸泡,对照组用去离子水.洗脱玻璃棒头部分泌物,做淋球菌分离培养,比较两组检出率.结果 (1)模拟雄鼠向雌鼠传播淋球菌实验中,实验组淋球菌培养阳性率低于对照组(第4天:10.0%比57.5%;第5天:7.5%比45.0%,均P＜0.01);实验组小鼠在接种后5～8 d PMN百分比显著低于对照组(均P＜0.05).(2)模拟雌鼠向雄鼠传播淋球菌实验中,第1、2天实验组玻璃棒头部分泌物淋球菌培养阳性率分别为8.3%和0,显著低于对照组(72.7%和45.5%,均P＜0.01).结论 长效抗菌材料通过形成物理抗菌膜,可以预防雌性小鼠生殖道淋球菌的感染或定植,同时能对雄性小鼠能起到液态安全套的作用.

Abstract：

Objective To explore a new method of preventing Neisseria gonorrhea (N.gonorrhoeae) by simulating intercourse in mice and verify the effect of a long-acting antibacterial material in the gonococcal infection in mice.Methods (1) Simulated male-female gonococcal transmission: the estradio-treated BALB/c female mice were inoculated with N.gonorrhoeae WHO-L via a vaginal route.In treated group,the mice received the long-acting antibacterial material via vaginal route while deionized water was administered in the control group.At Days 2-10 post-inoculation,the average number of polymorphonuclear cells (PMNs) was monitored in stained smears of vaginal cells from test mice.And the vaginal fluid was cultured for N.gonorrhoeae at Days 2,4,5 post-inoculation.N.gonorrhoeae genital tract infection was determined to compare the difference between the two groups.(2) Simulated female-male gonococcal transmission: a treated or sham glass rod was used instead of male mice's penis to simulate intercourse in gonococcus-infected female mice.The vaginal fluid on top of glass top was eluted and cultured for N.gonorrhoeae and the difference of relevance ratios compared.Results ( 1 ) In the experiment of simulated male-female gonococcal transmission,At Days 4 and 5,the positive rates of culture for N.gonorrhoeae of the tested group were lower than those of the control group,and there was significant difference between both groups ( 10.0% vs 57.5%,7.5% vs 45.0%,both P ＜0.01 ).The percentage of PMNs in the treated group was significantly lower than that of the control group at Days 5-8 post-inoculation (all P＜0.05 ).(2) In the experiment of simulated female-male gonococcal transmission,the positive rates of culture for N.gonorrhoeae from the vaginal fluid on top of glass top were 8.3% and 0 at Days 1 and 2 in the tested group respectively.And they were significantly lower than those of the control group (72.7% and 45.5% respectively,both P＜0.01 ).Conclusion After topical application in murine vagina,the longacting antibacterial material forms a layer of physically antibacterial molecular film to prevent the occurrence of N.gonorrhoeae genital tract infection or colonization in mice.It may be used as a liquid condom for male mice.     

Study of Candida Albicans Vaginitis Model in Kunming Mice

The model of vaginal candidiasis in Kunming mice was constructed in order to search for the optima construction conditions and provide an economic animal model of Candida albicans (C. albicans) vaginitis. Estrogen benzoate (E2) was given to mice at different concentrations ranging from 0.0 to 0.05 mg/mouse (4 levels) beginning 72 h prior to vaginal inoculation, then mice were in- oculated intravaginally with various concentrations of stationary-phase C. albicans blastoconidia (ATCC90028) (5 levels) in 20 μL of phosphate-buffered saline (PBS) in each E2 level. General state, scores of genital pathology, the hyphae and vaginal fungal burden (CFU) in vaginal lavage fluid, the hydrops rate of uterus and vaginal tissues for pathological section in mice were observed and ob- tained at day 2, 4, 7, 14 and 21 after inoculation. The results showed the infection rate in mice was related to the dosage of E2 and concentration of C. albicans blastoconidia. Additionally there was better cross-effect between the two treated factors. The infection rate was about 80% on the day 4, and could reach 100% on the day 7 until the end of experiment after inoculated intravaginally in groups of E2I3, E2 0.025 mg/mouse injected hypodermically and inoculated intravaginally with 5×104 C. albicans blastoconidia, and large amount of hyphae and blastoconidia could be observe in superfi- cial layer tissue and canal of vaginal by PAS. From the results in our experiment it was concluded that E2I3 was the optima construction condition in kunming mice.     

雌孕激素在构建小鼠生殖器疱疹动物模型中的比较研究

目的探讨用17β-雌二醇和孕酮构建小鼠生殖器疱疹动物模型的方法。方法实验前14d摘除雌性BALB/c小鼠卵巢,并经皮下注射17β-雌二醇、醋酸甲羟孕酮(长效孕酮)、孕酮或雌孕激素联合预处理,以生殖器单纯疱疹病毒(HSV-2)攻击小鼠生殖道,监测小鼠生殖周期、外阴病变和病毒滴度。结果孕酮预处理后小鼠进入动情间期并对HSV-2易感,长效孕酮可延长小鼠的动情间期至30d;雌二醇处理后小鼠进入动情期并对HSV-2不易感染;雌孕激素联合处理后小鼠进入动情期-动情间期,并对HSV-2易感染,表现明显的生殖器疱疹症状;雌孕激素剂量不影响易感性,也不影响小鼠感染程度和存活率。结论孕酮联合雌激素预处理,用低滴度HSV-2(333)感染,可以构建易感稳定的小鼠生殖器疱疹模型。     

淋球菌外膜蛋白NspA基因重组子的构建与表达

目的构建淋球菌标准株外膜蛋白NspA基因重组子,在大肠杆菌BL21（DE3）中诱导表达外膜蛋白NspA。方法提取淋球菌标准株基因组DNA,PCR扩增其NspA基因,插入表达质粒载体pET-30c（＋）中,构建pET-NspA重组子,转化表达宿主大肠杆菌BL21（DE3）,异丙基-β-D-硫代半乳糖苷（IPTG）诱导重组蛋白表达,SDS-PAGE和Western Blot检测表达蛋白。结果成功构建了淋球菌标准株的pET-NspA大肠杆菌表达重组子,经IPTG诱导表达后,该蛋白在大肠杆菌中高效表达。结论淋球菌外膜蛋白NspA在大肠杆菌中的成功表达,为研究该蛋白的免疫学性能、制备抗体和研制预防淋病的疫苗奠定了基础。     

Study of <Emphasis Type="Italic">Candida albicans</Emphasis> vaginitis model in Kunming mice

Summary The model of vaginal candidiasis in Kunming mice was constructed in order to search for the optima construction conditions and provide an economic animal model of Candida albicans (C. albicans) vaginitis. Estrogen benzoate (E₂) was given to mice at different concentrations ranging from 0.0 to 0.05 mg/mouse (4 levels) beginning 72 h prior to vaginal inoculation, then mice were inoculated intravaginally with various concentrations of stationary-phase C. albicans blastoconidia (ATCC90028) (5 levels) in 20 μL of phosphate-buffered saline (PBS) in each E₂ level. General state, scores of genital pathology, the hyphae and vaginal fungal burden (CFU) in vaginal lavage fluid, the hydrops rate of uterus and vaginal tissues for pathological section in mice were observed and obtained at day 2, 4, 7, 14 and 21 after inoculation. The results showed the infection rate in mice was related to the dosage of E₂ and concentration of C. albicans blastoconidia. Additionally there was better cross-effect between the two treated factors. The infection rate was about 80% on the day 4, and could reach 100% on the day 7 until the end of experiment after inoculated intravaginally in groups of E2I3, E₂ 0.025 mg/mouse injected hypodermically and inoculated intravaginally with 5 × 10⁴ C. albicans blastoconidia, and large amount of hyphae and blastoconidia could be observe in superficial layer tissue and canal of vaginal by PAS. From the results in our experiment it was concluded that E2I3 was the optima construction condition in kunming mice. CHEN Zhuo, female, born in 1965, Associate Professor This project was supported by the National “10th Five-Year” Key Technologies R&D (No.2004BA709B13-02)     

沙眼衣原体T细胞表位融合蛋白对小鼠生殖道沙眼衣原体感染的保护作用

目的：评价抗沙眼衣原体(Ct）感染的T细胞表位融合蛋白疫苗（H-ctm1）对小鼠生殖道感染的保护作用。方法：6～8周龄雌性C57BL/6小鼠分为3组：H-ctm1、热灭活Ct（HK-EBs）和磷酸盐缓冲液（PBS）免疫组（每组28只，其中9只用于衣原体感染包涵体数量的检测，10只用于组织病理学观察，9只用于输卵管积水情况分析）。3组分别用H-ctm1、HK-EBs和PBS免疫。通过阴道接种Ct感染小鼠，建立Ct感染小鼠生殖道的动物模型。接种前7 d皮下注射黄体酮以增加小鼠对Ct感染的敏感性。并通过该动物模型比较3组小鼠阴道分泌物中的Ct数量、阴道组织炎症病理积分及输卵管积水情况，评价H-ctm1抗沙眼衣原体感染的能力。结果：在阴道接种Ct后第3和6天，H-ctm1和HK-EBs两免疫组小鼠阴道分泌物中Ct数量比较差异均无显著性，但两组均明显少于PBS免疫组（P＜0.01）；在接种Ct后第9和18天，H-ctm1组小鼠阴道分泌物中Ct数量明显少于HK-EBs组（P＜0.01或P＜0.05）。在接种Ct后第6天，H-ctm1和HK-EBs组的炎症积分均明显低于PBS组（P＜0.01或P＜0.05）；在感染Ct后第12天， H-ctm1组的炎症积分明显低于HK-EBs组和PBS组（P＜0.05或P＜0.01）。在感染Ct后第40天，H-ctm1 和HK-EB_S组小鼠均未发生输卵管积水，而PBS组9只小鼠均发生单侧或双侧输卵管积水。结论：注射H-ctm1诱导小鼠产生较好的抗Ct感染的保护性免疫，且H-ctm1的免疫原性优于Ct灭活疫苗。     

荧光定量—聚合酶链反应检测性传播性疾病病原体结果分析

用荧光定量—聚合酶链反应 (FQ -PCR)检测 2 2 0 2份泌尿生殖道分泌物中的UU、CT、NG。了解性传播性疾病中解脲脲原体 (UU)、沙眼衣原体 (CT)以及淋病奈瑟菌 (NG)的感染情况。三种病原体中 ,UU的阳性率居于首位 ,明显高于NG的阳性率 ,而CT与NG的阳性率差异无显著性。混合感染阳性率不高 ,由UU或CT引起的混合感染率与由NG引起的混合感染率差异无显著性。     

Construtcion of Neisseria Gonorrhoeae Porin B Plasmid Recombinant and Its Expression in E.coli

Neisseria gonorrhoeaeis a common pathogenicmicroorganism which causes sex transmitted dis-ease . The porins ,the predominant proteins on thesurface of pathogenicNeisseria,form a family ofstructurally related proteins whose physiologicalrole is to allownutrients access into the cell . Theyalso play ani mportant role in pathogenesis ,gener-atingi mmunological specificity andinteracting withthe host cell .Neisseria gonorrhoeaeexpresses oneof the two alternative classes of porins PIA andPIB[1].…     

克淋灵经药粉吹入器给药治疗女性下生殖道淋病

目的研究克淋灵局部用药治疗女性下生殖道淋病的疗效。方法女性下生殖道淋病患者５６例，用阴道药粉吹入器将克淋灵药粉直接涂布于宫颈、穹窿和阴道内进行治疗（治疗组），并作体外细菌培养和药物敏感试验，同时随机选５０例女性下生殖道淋病患者肌注壮观霉素（淋必治），每次２ｇ，每日一次，共２次作为对照（对照组）。结果克淋灵经阴道药粉吹入器给药治疗女性下生殖道淋病治愈率１００％，明显高于对照组治愈率８０％（Ｐ＜０．０１）；克淋灵治疗组毒副反应发生率为０，明显低于对照组５０％（Ｐ＜０．０１）。克淋灵体外药物敏感试验１００％敏感，高于壮观霉素８０％敏感。结论克淋灵经阴道药粉吹入器给药治疗女性下生殖道淋病是一种疗效显著、毒副反应小、经济、方法简单易行、值行推广的治疗女性下生殖道淋病的新方法