UU1、UU3和UU4致昆明小鼠下生殖道致病性的研究

目的:研究解脲脲原体(Ureaplasma Urealyticum,UU)的第1血清型(UU1)、第3血清型(UU3)和第4血清型(UU4)对昆明小鼠下生殖道的致病性。方法:将100只昆明小鼠随机分为5组:空白组、雌二醇组、UU1组、UU3组和UU4组,每组20只;空白组和雌二醇组为对照组,UU1组、UU3组和UU4组为实验组;雌二醇组和实验组雌二醇化后1周,将UU1、UU3和UU4分别接种至UU1、UU3和UU4组小鼠泌尿生殖道,将液体培养基接种至空白组和雌激素组。2周后检测各组小鼠泌尿生殖道的IL-8、SIgA、TNF-α并取小鼠子宫组织行病理检查。结果:实验组小鼠UU接种都获得成功。IL-8、SIgA、TNF-α检测结果显示:实验组和对照组比较有显著性差异;UU4组和UU1组及UU3组比较有显著性差异;空白组和雌二醇组比较无显著性差异;UU1组和UU3组比较无显著性差异。病理显示:UU1、UU3和UU4组昆明小鼠子宫组织有炎细胞浸润,UU4组小鼠宫颈病变更明显。对照组小鼠子宫未见明显改变。结论:UU1、UU3和UU4可能导致昆明小鼠下生殖道产生病理反应;UU4可能短期致病力更强。     

泌尿生殖道支原体培养及耐药性差异分析

目的检测本地区人群泌尿生殖道溶脲脲原体（Uu）和人型支原体（Mh）感染的发生率;分析其对9种抗生素的药物敏感性差异,用以指导临床的抗生素治疗。方法采用法国生物梅里埃Mycoplasma IST2试剂盒对支原体进行分离培养鉴定和药物敏感性试验;分析2008—2013年本地区支原体感染患者耐药性的差异。结果在本地区8168例疑似NGU的患者中检出支原体2564例,阳性检出率为31．39％,其中仅uu感染的患者1833例（22．44％）,Uu和Mh混合感染的患者623例（7．63％）,仅Mh感染的患者108例（1．32％）。仅uu感染的患者对PRI、JOS、DOT、TET、CLA、AZI、ERY、OFL和CIP九种抗生素的敏感率分别为99．8％、99．4％、96．3％、91．9％、90．O％、89．7％、69．7％、4．O％和0．5％;Uu和Mh混合感染的患者对9种抗生素的敏感率分别为96．3％、96．1％、94．8％、79．5％、12．0％、11．7％、1．4％、1．0％和0．3％;仅Mh感染的患者对9种抗生素的敏感率分别为100％、100％、100％、90．7％、0％、0％、O％、14．8％和13．9％。结论本地区泌尿生殖道支原体感染以Uu感染最为常见,Uu和Mh混合感染次之,Mh感染最为少见;对支原体的抗生素治疗首选PRI、JOS、DOT;TET、AZI、CLA可适当使用,而ERY、OFL、CIP则不宜选用。     

男性泌尿生殖道支原体感染及药敏分析

探讨男性尿道炎患者支原体感染率及其敏感性,为临床治疗支原体感染导致的疾病提供合理用药的参考依据。采用海泰生物的史原体分离鉴定试剂盒,取前列腺液或尿道分泌物,对1116份疑为泌尿生殖系统感染的男性患者进行了支原体检测和抗生素敏感性检测。1116份男性患者标本中共分离出274株支原体,242株Uu对美满霉素、强力霉素、环雨沙星、司帕沙星、氧氟沙星、阿齐霉素、罗红霉素、交沙霉素、克拉霉素、壮观霉素的敏感率分别为95．1％、94．6％、17％、70．3％、31．1％、76％、77．7％、92．6％、83．5％、43．8％。30椿Uu＋Mh对美满霉素、强力霉素、环丙沙星、司帕沙星、氧氟沙星、阿齐霉素、罗红霉素、交沙霉素、克拉霉素、壮观霉素的敏感率分别为70％、66．7％、20％、40．1％、26．8％、23．4％、23．4％、50％、23．3％、20％。泌尿生殖道支原体的耐药性监测对指导临床治疗具有重要意义,可以根据药敏结果指导临床合理使用抗菌药物。     

解脲脲原体药物敏感性变迁研究

目的:探讨上海地区2008与2012年泌尿生殖道解脲脲原体(Ureaplasma urealyticum,UU)耐药性变化,为临床合理用药提供参考。方法:采用自制UU药物敏感检测试剂对2008年450例和2012年459例患者的标本进行检测,观察UU阳性情况及UU对交沙霉素、氧氟沙星、阿奇霉素和强力霉素的药物敏感性。结果:2008年分离到150例UU阳性标本和2012年分离到134例UU阳性标本分别对交沙霉素、强力霉素、氧氟沙星和阿奇霉素等4种抗菌素的敏感率有显著性差异;2008年46例和2012年38例男性患者阳性标本中分离的UU分别对交沙霉素和阿奇霉素敏感率有显著性差异;对强力霉素和氧氟沙星敏感率无显著性差异。2008年104例和2012年96例女性患者阳性标本中分离的UU分别对交沙霉素、强力霉素、氧氟沙星和阿奇霉素等4中抗菌素的敏感率都有显著性差异。结论:5年间UU药物敏感性发生了变迁;临床医师应该关注本地区药物敏感性变迁,合理选用抗生素。     

Emerging role for Ureaplasma parvum serovar 3: Active infection in women with silent high-risk human papillomavirus and in women with idiopathic infertility

Recently, there are controversial opinions on the presence of Mycoplasmas/Ureaplasmas as colonizers or pathogens, and on the use of a targeted therapy. This study aimed to characterize Mycoplasmas/Ureaplasmas infections in reproductive age women, including the acquisition of sexually transmitted (ST) pathogens and poor birth outcomes. A total of 646 healthy Italian women fulfilled the inclusion criteria including 521 infertile women, 65 pregnant women, and 60 fertile women with identified risk factors and symptomatic for vaginitis/cervicitis. Multiplex and quantitative molecular techniques and direct automatic DNA sequencing were performed to assess the genome structure of Mycoplasma/Ureaplasma species and ST infected pathogens. Ureaplasma parvum serovar 3 represented the predominant colonizer of the urogenital tract of this series and the unique species significantly associated with ST pathogens coinfection (p < 0.01). U. parvum load >10⁴ bacteria/ml, suggestive of active infection, has been measured only in asymptomatic high-risk human papillomavirus infected women (24.3%) and in 40% of women with idiopathic infertility. To note, 16% of the follicular fluid from these idiopathic women resulted infected with U. parvum. In conclusion, the present study focused the attention on U. parvum serovar 3 as emerging microorganism in sexually active women that may have the benefit of targeted therapy.     

生殖道衣原体、支原体感染与盆腔炎症的相关性分析

目的:探讨女性生殖道衣原体、支原体感染发生与盆腔炎症的相关性,并为相应人群提出相应的预防和治疗措施。方法:对我院2012年1月到2013年5月诊治的盆腔炎患者280例及60名健康妇女进行了衣原体、支原体的培养,采用试剂盒进行检查,并进行药敏实验,分析比较两组生殖道衣原体和支原体感染情况。结果:盆腔炎症组中衣原体感染的检出率为58.5%,支原体感染率为26.2%,衣原体合并支原体感染率为12.4%。健康妇女组衣原体感染、支原体感染及衣原体合并衣原体感染的检出率分别为:9.1%,6.2%和4.9%。两组的差异有统计学意义(P0.05)。在盆腔炎症患者组中,30岁人群中单纯衣原体感染和单纯支原体感染检出率分别为51.3%和26.4%,均要明显高于30岁以上的人群,差异有统计学意义(P0.05)。结论:盆腔炎症与生殖道衣原体,支原体感染有密切的相关性,盆腔炎的发病可能与生殖道衣原体、支原体感染有关,针对临床上盆腔炎患者应密切关注生殖道支原体、衣原体的感染问题。     

Vaginal Secretion Epithelium Count As a Prognostic Indicator of High Abundance of Ureaplasmas in Women with a Normal Nugent Score

Genital tract ureaplasma infections are associated with numerous complications, ranging from inflammation, through infertility, to problematic pregnancy. In the course of ureaplasma infection, the risk of human papillomavirus infection increases. Diagnostic tests for urea-plasma infections are not always carried out, especially in women with the normal Nugent test results. The study attempts to check whether it is possible to find a prognostic indicator that could suggest a high abundance of ureaplasmas (≥ 10⁴ CFU/ml) at the stage of the initial examination of vaginal discharge. Such a prognostic factor could qualify women for further tests to detect infections with these atypical bacteria. Six hundred twenty-seven white women with a score of 0–3 on the Nugent scale were tested, including 322 patients with a high abundance of ureaplasmas (≥ 10⁴ CFU/ml) and 305 who tested negative for these bacteria. Ureaplasma infections were detected statistically significant in women who had few or no epithelial cells in the genital swab specimens compared to the results obtained for women with numerous or very numerous epithelial cells (p < 0.001). The risk of the high density of ureaplasmas was 38.7% higher with fewer or no epithelial cells than with high numbers. In patients aged 18–40 years with few or no epithelial cells, a high density of ureaplasmas (≥ 10⁴ CFU/ml) was observed significantly more frequently (p = 0.003). Determining the number of epithelial cells in Gram-stained slides may be the prognostic indicator of ureaplasma infection. Testing for genital ureaplasma infection should be considered, especially in women of childbearing age (18–40 years), even if the Nugent test value is normal and pH ≤ 4.6.     

酶联免疫吸附试验夹心法检测解脲脲原体方法的建立

目的为了提高解脲脲原体(Ureaplasma urealyticum,UU)检测的快速性。方法用酶联免疫吸附试验(ELISA)夹心法检测UU抗原并与传统的培养法相比较。结果ELISA夹心法敏感度为92.6%,特异度为97.4%,最低能够检测出蛋白含量为5～10ng/ml的UU抗原。结论ELISA夹心法是一种敏感、方便、快捷、适合大规模标本检测解脲脲原体的方法。     

TaqMan探针双重荧光定量PCR同时检测解脲支原体和巨细胞病毒

目的探讨双重荧光定量PCR技术的优化条件,建立基于TaqMan探针技术荧光定量法检测同时检测解脲支原体和巨细胞病毒的新方法。方法分别采用普通定性PCR扩增母婴垂直传播常见的病原体（解脲支原体和巨细胞病毒）测序鉴定,然后分别采用TaqMan探针的单重和双重定量PCR技术对解脲支原体和巨细胞病毒同时定性定量检测。结果解脲支原体和巨细胞病毒单种定性PCR检测均为阳性,TaqMan探针单重和双重定量PCR检测解脲支原体和巨细胞病毒阳性率和特异性均为100％,相同样品TaqMan探针单重、双重定量PCR分别检测的结果符合率100％。结论TaqMan探针双重荧光定量PCR技术可同时检测两种靶分子,结果可靠,应用前景广阔。     

解脲脲原体脂质相关膜蛋白通过激活核因子κB调控诱导性一氧化氮合酶在小鼠巨噬细胞中的表达

为探讨解脲脲原体(Uu)的脂质相关膜蛋白(LAMPs)诱导小鼠巨噬细胞表达诱导性一氧化氮合酶(iNOS)的分子机制,从解脲脲原体提取的脂质相关膜蛋白,刺激小鼠巨噬细胞,以RT_PCR、Western blot等方法分析iNOS的表达及NO的产生;用细胞免疫化学、间接免疫荧光及Western blot等方法检测核因子κB(NF_κB)的激活,另外检测了NF_κB的特异性抑制剂二硫代氨基甲酸吡咯烷(PDTC)和蛋白酶抑制剂放线菌酮(CHX)对iNOS的表达及NF_κB激活的影响。结果表明,解脲脲原体的LAMPs通过激活NF_κB诱导小鼠巨噬细胞表达iNOS的mRNA和蛋白,且能以时间和剂量依赖方式刺激小鼠巨噬细胞产生NO,NF_κB的抑制剂PDTC或蛋白酶抑制剂放线菌酮(CHX),可抑制NF_κB的激活及iNOS的表达。由于解脲脲原体的脂质相关膜蛋白通过激活NF_κB诱导小鼠巨噬细胞表达iNOS和产生NO,因而可能是一个重要的致病因素。