Trifluoperazine inhibits the infectivity of Chlamydia trachomatis for McCoy cells

Abstract Trifluoperazine (TFP), an inhibitor of the Ca²⁺-binding protein calmodulin, was used to study the infectivity of Chlamydia trachomatis for McCoy cells. TFP inhibited the number of chlamydial inclusions and the chlamydia-dependent amino acid incorporation when added within 9 h after inoculation with chlamydiae. However, TFP did not affect the attachment of chlamydiae to the cells or the protease-removable fraction of cell-bound chlamydiae.
These results suggest that an early step in the intracellular development of chlamydiae, partly coinciding with the elementary body-reticulate body conversion, is sensitive to TFP and that clathrin coats are not crucial in the ingestion of chlamydiae by McCoy cells.     

生殖道感染与宫颈癌及宫颈癌前病变的相关性研究

目的:探讨生殖道感染与宫颈癌、宫颈癌前病变的相关性及危险因素分析。方法:选取我院收治的185例宫颈癌及宫颈癌前病变患者和同期206例健康体检者分为两组,对人乳头瘤病毒(HPV)、沙眼衣原体(CT)、细菌性阴道病(BV)、阴道滴虫进行检测,观察分析相应病原微生物导致的生殖道感染与宫颈癌及宫颈癌前病变的关系,同时根据HPV不同基因型在宫颈癌及宫颈癌前病变中的致癌作用和程度,判定危险程度。结果:宫颈癌及宫颈癌前病变患者上述指标感染检出率明显高于对照组,差异具有统计学意义(P=0.000、0.001、0.000、0.037),其中高危HPV感染率随宫颈上皮内瘤变级别的升高而呈明显上升趋势,CINⅠ～Ⅲ级感染率分别为57.1%、78.6%和82.9%,宫颈癌感染率最高达91.1%,高危HPV为高危因素。结论:生殖道HPV、CT、BV、滴虫感染与宫颈癌及CIN存在必然或一定相关性,加强对妇女生殖道病原体感染的重视和检测对防治宫颈癌及CIN具有积极意义。     

沙眼衣原体感染诱导小鼠生殖道局部促炎性细胞因子的表达情况

建立小鼠生殖道沙眼衣原体感染模型,观察小鼠生殖道局部促炎性细胞因子的表达。将小鼠生物型沙眼衣原体C. muridarum 1＆#215;104 IFU阴道接种于C57B6背景雌性小鼠,取感染后阴道拭子做沙眼衣原体培养,计算IFU,监测小鼠感染和病原体清除情况;80 d后处死小鼠,检测子宫输卵管病理改变;ELISA检测感染过程中小鼠生殖道促炎性细胞因子IL-1α、IL-6、MIP-2和TNF-α产生情况。小鼠感染在第3至第15天维持较高水平,然后病原体被逐渐清除,整个病程约3~5周;病理检测显示子宫输卵有严重炎症、管腔扩张积水,狭窄等;于感染后第3天检测到局部IL-1α、IL-6、MIP-2分泌,第7天达高峰,然后逐渐下降至正常水平（ IL-6于11 d恢复正常,IL-1α和 MIP-2于15 d恢复正常）。 TNF-α仅在第7天检测到高水平表达。相对于TNF-α和IL-6,IL-1α和MIP-2维持时间较长。成功建立沙眼衣原体感染小鼠生殖道模型,沙眼衣原体急性感染可诱导小鼠生殖道局部分泌IL-1α、IL-6、MIP-2和TNF-α。     

肺炎支原体和沙眼衣原体致儿童急性下呼吸道感染的研究

本文旨在研究儿童社区获得性急性下呼吸道感染(ALRTI)中肺炎支原体(MP)和沙眼衣原体(CT)的感染特征。采用实时荧光定量聚合酶链反应(PCR)检测2006年10月～2008年2月因ALRTI收入复旦大学附属儿科医院的患儿呼吸道标本中MP和CT的DNA,并分析2种病原体感染患儿的临床特征与实验室检查结果。在1312份深部鼻咽分泌物标本中,MP和CT检出率分别为7.85%(103/1312)和2.97%(39/1312)。MP在5岁以上患儿中的检出率为33.33%(30/90),而CT在3个月以内患儿中的检出率为6.28%(31/494)。MP感染后易出现40℃以上高热,较少发生发绀与重症呼吸道感染;白细胞计数常无明显升高,但C反应蛋白升高较多见。CT感染后40℃以上高热和重症呼吸道感染少见,C反应蛋白升高也较少见。结果提示,在5岁以上儿童的社区获得性ALRTI中,MP是重要的病原体;而在3个月以内儿童中,CT为常见病原体之一。     

孕晚期胎膜早破孕妇产道无乳链球菌定植与新生儿感染的相关性分析

目的 分析孕晚期胎膜早破妇女产道无乳链球菌（SA）定植与新生儿感染之间的相关性。方法 筛选2014年1月—2017年3月我院产科收治的孕晚期胎膜早破孕妇589例作为观察组,无胎膜早破的正常孕晚期孕妇261例作为对照组,对两组孕妇宫颈拭子进行细菌培养,并对其分娩的新生儿咽拭子进行细菌培养,记录两组孕妇胎膜早破合并SA阳性结果孕妇数与新生儿感染人数。结果 胎膜早破孕妇宫颈拭子SA阳性率为16.47%（97/589）,无胎膜早破的正常妊娠晚期妇女SA阳性率为6.13%（16/261）（P＜0.01）;观察组SA阳性孕妇新生儿感染发生率6.18%（6/97）,显著高于本组SA阴性孕妇4.67%（23/492）（P＜0.05）;观察组破膜时间≥24 h孕妇其新生儿感染率为62.07%（18/29）,高于本组破膜时间＜24 h孕妇的新生儿感染率27.94%（19/68）（P＜0.01）;观察组产程≥24 h孕妇其新生儿感染率42.42%（14/33）,高于本组破膜时间＜24 h孕妇的新生儿感染率17.19%（11/64）（P＜0.05）。结论 孕晚期孕妇产道SA感染导致胎膜早破风险增加,新生儿感染率升高,有必要对SA阳性孕妇及早干预治疗,尽量减少胎膜早破,缩短产程。     

Strategies for optimizing DNA hybridization on surfaces

Specific and predictable hybridization of the polynucleotide sequences to their complementary counterparts plays a fundamental role in the rational design of new nucleic acid nanodevices. Generally, nucleic acid hybridization can be performed using two major strategies, namely hybridization of DNA or RNA targets to surface-tethered oligonucleotide probes (solid-phase hybridization) and hybridization of the target nucleic acids to randomly distributed probes in solution (solution-phase hybridization). Investigations into thermodynamic and kinetic parameters of these two strategies showed that hybridization on surfaces is less favorable than that of the same sequence in solution. Indeed, the efficiency of DNA hybridization on surfaces suffers from three constraints: (1) electrostatic repulsion between DNA strands on the surface, (2) steric hindrance between tethered DNA probes, and (3) nonspecific adsorption of the attached oligonucleotides to the solid surface. During recent years, several strategies have been developed to overcome the problems associated with DNA hybridization on surfaces. Optimizing the probe surface density, application of a linker between the solid surface and the DNA-recognizing sequence, optimizing the pH of DNA hybridization solutions, application of thiol reagents, and incorporation of a polyadenine block into the terminal end of the recognizing sequence are among the most important strategies for enhancing DNA hybridization on surfaces.     

性病患者泌尿生殖系病原体感染分析

目的 结合临床分析医院性病门诊具有泌尿生殖道刺激症状及异常分泌物患者的淋球菌(NG)、沙眼衣原体(CT)和解脲脲原体(UU)的感染状况,为临床治疗提供依据.方法 对门诊283例患者标本作NG、CT、UU检测,其中UU、CT采用新型快速试剂盒检测;NG检测采用分泌物涂片、染色、镜检,可疑者再用培养法确证.结果 三种病原体总感染率为62.19％(176/283),单一感染占39.22％(111/283),混合感染占22.97％(65/283),混合感染中以CT+ UU感染(13.07％)为主;男、女感染率分别为59.65％和66.07％,患者年龄集中在21 ～40岁,以31 ～40岁组阳性病例数最多(35.34％);职业分布:病例数最多的是无业人员(45.58％),其次为工人、干部.非淋菌性尿道(宫颈)炎(NGU)和淋病(GU)的阳性检出率分别为46.64％和15.54％.结论 NGU和GU的的感染以性成熟期患者为主,NGU发病率高,应引起高度重视.     

用核酸杂交法检查北京地区幼儿园正常儿童尿中人巨细胞病毒DNA

以克隆的人巨细胞病毒(Human Cytomegalovirus,简称HCMV)DNA片段做探针,用核酸杂交方法检查了50例北京地区幼儿园正常儿童的尿标本。查出HCMV DMA阳性者24例,占48％。在两个群体和两个年龄组之间,阳性检出率有显著性差异。利用探针可查出同源DNA5～10pg,而不与其它病毒或细胞DNA杂交。标记不同的探针,使用不同的固定方法,其敏感性不同。此法可在分子水平上查出HCMV基因,比分离病毒更迅速敏感。     

基因扩增产物的固相杂交-酶联显色方法的建立

建立基于基因扩增技术的简便、快速的病毒核酸定量检测方法．将标记有生物素的寡核苷酸引物所扩增的病毒基因产物,与通过共价键结合在微孔反应板上的特异性探针进行快速杂交,然后通过辣根过氧化物酶标记的抗生物素进行酶联显色,读取光密度值．应用本方法对血清中乙型、丙型肝炎病毒核酸定量检测,灵敏度分别可达１－５拷贝／反应．此方法简便、快速、特异性好、敏感性高、半定量指标客观,可广泛应用于肝炎病毒感染的临床诊断和疗效评价．     

Stress-induced protein synthesis in Chlamydomonas reinhardtii

Deoxyribonucleic acid base composition, deoxyribonucleic acid-deoxyribonucleic acid hybridization, and biochemical studies were performed on some enterococci from clinical sources of uncertain taxonomic position. Our results indicate that 6 human strains, a single clinical isolate and a strain from bovine mastitis are genetically distinct from each other and all other previously described Enterococcus species and constitute three new species, for which the names Enterococcus raffinosus, Enterococcus solitarius and Enterococcus pseudoavium are proposed.     

实时荧光聚合酶链反应检测性病患者泌尿生殖道沙眼衣原体的临床分析

采用实时荧光聚合酶链反应（PCR ）检测性病患者泌尿生殖道沙眼衣原体,进行临床和实验室分析。采集380例男女性病患者泌尿生殖道分泌物,进行多形核白细胞数检测和荧光PCR。结果显示,216例男性患者中,尿道多形核白细胞数≥5个者占92．59％,荧光PCR检测沙眼衣原体阳性62例,阳性率为28．70％;沙眼衣原体合并淋病奈瑟菌感染30例,合并感染率为13．89％。164例女性患者中,87例宫颈管内多形核白细胞数≥10个（占53．05％）,荧光PCR检测沙眼衣原体阳性33例（占20．12％）。结果提示,性病门诊开展实时荧光PCR检测沙眼衣原体可提高检测阳性率和控制沙眼衣原体传播。     

沙眼衣原体感染活化NOD1信号通路的研究

目的:探索沙眼衣原体(Chlamydia trachomatis,Ct)持续感染状态下,NOD1、IL-6及STAT3分子的表达情况和相互关系。方法:利用HeLa229细胞或STAT3基因沉默的HeLa229细胞,分别建立沙眼衣原体的急性感染和持续感染模型;应用Western Blot及ELISA等方法检测不同感染状态下STAT3及NOD1蛋白表达水平以及细胞因子IL-6的分泌水平。结果:HeLa229细胞在Ct感染状态下,STAT3和NOD1以及IL-6表达水平均升高,且于持续感染状态下的升高较急性感染状态下的升高更明显;沉默STAT3基因后,Ct感染的细胞NOD1及IL-6的表达水平下降明显。结论:HeLa229细胞在Ct持续感染状态下,STAT3能上调NOD1及IL-6表达水平,上述分子间存在NOD1-IL-6-STAT3正反馈信号通路。     

肺炎衣原体HEP—2培养分离及其抗体的MIF研究

肺炎衣原体是一种引起肺炎及呼吸道感染等的新型病原体。本研究采用ＨＥＰ－２培养从８例呼吸道感染者新鲜痰液中初步分离出肺炎衣原体。此外,采用微量免疫光试验（ＭＩＦ）对９６例呼吸道感染者与４８例健康献血员的血清学分析表明：８３．４％的患者肺炎衣原体ＩｇＧ抗体阳性,与对照组比较,统计学上有非常显著差异。提示这些患者呼吸道感染中,肺炎衣原体可能起较大作用。     

Rapid detection of human rotavirus using colorimetric nucleic acid sequence-based amplification (NASBA)-enzyme-linked immunosorbent assay in sewage treatment effluent

A colorimetric nucleic acid sequence-based amplification-enzyme-linked immunosorbent assay (NASBA-ELISA) was developed for rapid detection and identification of human rotavirus. Oligonucleotide primers targeting gene 9 encoding a serotype-specific antigen VP7 were selected and used for the amplification of viral RNA by the isothermal NASBA process, resulting in the accumulation of biotinylated RNA amplicons. Amplicons were hybridized with a specific amino-linked oligonucleotide probe covalently immobilized on microtiter plates. The DNA-RNA hybrids were colorimetrically detected by the addition of streptavidin-peroxidase conjugate and tetramethylbenzidine substrate. Using the NASBA-ELISA system, as little as 0.2 PFU (4 x 10(1) PFU ml(-1)) and 15 PFU (3 x 10(3) PFU ml(-1)) of rotavirus were detected within 6 h in spiked MQ water and sewage treatment effluent respectively. No interference was encountered in the amplification and detection of rotavirus in the presence of non-target RNA or DNA. Moreover, the presence of non-target bacteria and virus does not generate any non-specific signal, confirming the specificity of the developed NASBA-ELISA system and its effectiveness in specifically detecting rotavirus. The NASBA-ELISA system offers several advantages in terms of sensitivity, rapidity and simplicity. This technique should be readily adaptable for detection of other RNA viruses in both foods and clinical samples.     

儿童泌尿系感染的病原学分析

探讨目前儿童泌尿系感染病原体的变化趋势,为临床治疗提供实验依据。分析2008年1月至2009年10月住院治疗的357例尿细菌培养、支原体体外培养、衣原体检测阳性的泌尿系感染患儿病原体的分布情况。结果显示,尿细菌培养和支原体体外培养、衣原体检测前未应用过抗生素的患儿其阳性率为82.2%,而应用过抗生素的患儿其阳性率为31.8%,两者相比具有统计学意义(P0.01)。在检测的357例阳性标本中,革兰阴性杆菌占74.7%,其中以大肠埃希菌为主,占46.2%;革兰阳性球菌占14.8%,其中肠球菌占10.9%;真菌占3.1%,支原体占4.8%,衣原体占2.5%。临床要密切关注儿童泌尿系感染病原体的分布变迁情况,以便于为临床的诊断和治疗提供可靠的实验依据。