结核分枝杆菌药物敏感性表型检测的研究进展

全球耐药结核病疫情依旧严峻,我国尤甚,结核分枝杆菌药物敏感性表型检测在耐药结核病的诊断中仍占有重要地位,本综述论述了近年来多种表型药敏检测方法及国内外研究进展,讨论汇总了各种药敏试验方法的敏感度、特异度及各方法的优缺点,为临床及实验室根据不同需求选择药敏方法提供依据。     

无创中心动脉压和血管弹性功能检测的合理应用

本文介绍了目前无创中心动脉压和动脉弹性功能检测方法的基本原理及优缺点,提出如何合理选择及应用这些检测方法的建议,并指出其中有些检测方法目前仍限于研究阶段,但随着研究资料充实,有可能在未来几年成为心血管危险分层及监测药物疗效的有用指标。     

血吸虫病抽样调查居民感染率及感染者例数推算方法的探讨

目的：探讨血吸虫病抽样调查居民感染率及感染者例数的推算方法。方法：结合多重分层整群随机抽样方法的特点，应用血吸虫病流行病学和医学统计学原理，对居民感染率及感染者例数的统计方法进行理论推导。结果：得出各流行省及其不同类型未控制流行区的居民感染率及感染者例数的１套理论计算公式。结论：推算方法简明、合理，有实际应用价值。     

分子生物学方法用于研究血吸虫种株遗传变异及分类

分子生物学方法研究血吸虫种株遗传变异及分类是最直接而可靠的方法。它弥补了以往方法的不足,使得对血吸虫种株的研究更加科学精确。本文就DNA为基础的各种分子生物学方法简要原理及其在血吸虫种株遗传变异及分类研究中的应用,作一概述。     

双PAP法显示胃肠道促胃液素释放肽细胞

目的 研究胃窦及十二指肠粘膜中促胃液素释放肽(GRP)细胞的免疫细胞化学染色方法,并评价其效果。 方法 PAP法及双PAP法。 结果 GRP细胞主要位于胃腺及十二指肠腺管中下部。双PAP法可大大提高染色阳性强度。 结论 双PAP法是显示胃肠道GRP细胞的较好方法。     

Uhl畸形诊断与治疗研究进展

Uhl畸形是一种罕见的先天性心脏病，Uhl畸形患儿预后较差，手术治疗为目前主要治疗方法。现回顾既往相关文献，总结Uhl畸形患儿临床表现、诊断方法及手术治疗的方法及疗效，探讨Uhl畸形患儿的最佳诊治方法。     

用微量全血彗星试验观察砷中毒病区人群DNA损伤

目的 将微量全血彗星实验方法及微量血样保存方法在地方性砷中毒病区进行实践，以验证该方法的可行性。方法 采集病区人群末梢微量血样及相应的水样及尿样，观察人群DNA损伤程度与总砷摄入量及尿砷之间的相关关系以及不同人群DNA损伤程度之间的差别。结果 没有发现总砷摄入量及尿砷与DNA损伤程度之间的相关关系，但砷中毒病人的DNA损伤程度明显高于非病人及可疑病人。结论 将微量血样保存方法及微量全血彗星实验用于地方性砷中毒病区的人群监测可行。     

裂隙灯下邮票式剔除法剔除角膜铁屑异物278例临床体会

目的探讨裂隙灯下剔除角膜铁屑异物及锈环的方法及疗效。方法对278例(296眼)角膜铁屑异物及形成锈环的患者表麻后在裂隙灯下用邮票式剔除法一次性剔除异物及锈环,观察临床疗效。结果本组病例均一次性将异物及锈环完整取出,全部治愈,未出现感染及并发症,无锈斑残留,遗留角膜薄翳32例。结论裂隙灯下邮票式剔除角膜铁屑异物是一种对角膜损伤小,安全有效的剔除方法。     

起搏器囊袋破溃9例分析

目的：探讨起搏器囊袋破溃的常见原因及合理处理方法。方法：回顾分析890例心脏永久性起搏器植入术患者，包括植入心脏复律除颤器及三腔起搏器患者的资料，其中9例患者术后发生起搏器囊袋破溃。分析上述患者的临床情况及处理过程，寻找引起起搏器囊袋破溃的常见原因及合理处理方法。结果：9例患者均为亚急性或慢性囊袋破溃，其中6例患者囊袋分泌物细菌培养阳性，大部分为表皮葡萄球菌。经在将起搏器移至对侧及抗感染治疗后破溃囊袋良好愈合。结论：永久起搏器术后囊袋破溃患者的常见原因是囊袋感染。合理的处理方法是抗感染及将原起搏器及电极取出，并植入对侧相应部位。     

偏瘫型偏头痛30例的临床分析

目的分析偏瘫型偏头痛主要临床症状及诊断、治疗方法。方法回顾性分析30例偏瘫型偏头痛患者的临床资料，对其临床症状表现及治疗方法进行探讨。结果30例患者经镇痛药、镇静药及血管扩张药治疗后痊愈且至今未复发。结论偏瘫型偏头痛症状表现特殊，可给予镇静药、镇痛药及血管扩张剂进行治疗和预防复发。     

河南省某一淡水养殖环节中非O1/O139群霍乱弧菌毒力基因及分子分型分析

目的 了解河南省淡水养殖环节中非O1/O139群霍乱弧菌毒力基因分布及分子分型情况。方法 对河南省淡水养殖环节中50株非O1/O139群霍乱弧菌和3株病人来源菌株进行全基因测序,利用PubMLST-Vc数据库分析其序列分型(ST),利用最小生成树关系图分析进化关系,通过VFDB数据库获得其毒力基因分布。结果 来源于淡水养殖环节和来源于病人的非O1/O139群霍乱弧菌53株菌株均具有黏附、趋化运动、抗吞噬、毒素及酶类等功能的8类毒力相关因子基因,缺失辅助定植、毒素共调、分泌等功能的毒力相关因子基因和副霍乱肠毒素等4个毒素基因。部分毒力相关因子或部分菌株的毒力相关因子毒力基因不全。与3株来源于病人的菌株相比,二者毒力因子基因携带情况相近,除MSHA Ⅳ型菌毛毒力因子mshA基因、荚膜多糖wbuB基因、wbfY基因、rmlB基因、血红素受体hutA基因及Ⅲ型分泌系统vscC2和vcrD2基因外,部分菌株二者携带相同的毒力因子基因。53株非O1/O139群霍乱弧菌分属19个ST型,ST4和ST5是优势ST型,养殖环节来源的菌株与病人来源的菌株分属不同的ST型。19个ST型等位基因位点变异差异数在1~7个,其中分离自淡水养殖环节的非O1/O139群霍乱弧菌菌株17个ST型等位基因位点变异差异数在1~7个,病人来源的非O1/O139群霍乱弧菌菌株2个ST型与分离自淡水养殖环节的非O1/O139群霍乱弧菌菌株ST1、ST2、ST6和ST10属同一簇,与ST1有6个等位基因位点存在差异。结论 河南省淡水养殖环节非O1/O139群霍乱弧菌菌株MLST分型多样化,携带多种毒力相关因子,不同来源菌株携带的毒力基因相同,虽然分属不同的ST型,但还是存在食品安全风险,提醒有关部门采取措施进行防控。     

云南省非O1/O139群霍乱弧菌分子特征分析

目的非O1/O139群霍乱弧菌能够引起人类急性腹泻性疾病,但与O1群和O139群相比,人们往往容易忽视其危害性。因此,我们对分离于云南省的31株非O1/O139群霍乱弧菌开展了分子特征分析。方法采用纸片法(K-B)开展抗生素敏感试验;多聚酶链反应(PCR)扩增检测毒力基因;同时,我们对菌株进行了脉冲场凝胶电泳(PFGE)和多位点序列分析(MLST)分子分型研究。结果药敏实验结果显示,67.74%的菌株对利福平耐药,对萘啶酸和复方新诺明的耐药率为29.03%,所有的菌株对庆大霉素和环丙沙星敏感;PCR结果显示,所有菌株的ompW基因为阳性,87.10%的菌株hly基因为阳性,25.81%的菌株rstREl tor为阳性,16.13%的菌株rstRClassical和tcpAEl tor为阳性,而CT、rfbO1和rfbO139基因全为阴性;PFGE结果显示,31株非O1/O139群霍乱弧菌呈现出高度的离散趋势,几乎没有相同带型的菌株出现;在MLST结果分析中,发现了1个gyrB新的等位基因,mdh基因发现了4个新的等位基因,metE基因发现了6个新的等位基因,pntA中发现了2个新的等位基因,purM中发现了3个新等位基因,pyrC中发现了4个新等位基因。经过排列组合后,共发现了17个新的霍乱弧菌ST型别(ST273-ST289)。结论非O1/O139群霍乱弧菌呈现出高度的多样性特征,同时,云南省的非O1/O139群霍乱弧菌具有一定的地域特点,丰富了现有的霍乱弧菌分子分型数据库。     

Genomic characterization of non-O1, non-O139 Vibrio cholerae reveals genes for a type III secretion system

Non-O1, non-O139 Vibrio cholerae can cause gastroenteritis and extraintestinal infections, but, unlike O1 and O139 strains of V. cholerae, little is known about the virulence gene content of non-O1, non-O139 strains and their phylogenetic relationship to other pathogenic V. cholerae. Comparative genomic microarray analysis of four pathogenic non-O1, non-O139 strains indicates that these strains are quite divergent from O1 and O139 strains. Genomic sequence analysis of a non-O1, non-O139 strain (AM-19226) that appeared particularly pathogenic in experimental animals suggests that this strain carries a type III secretion system (TTSS) that is related to the TTSS2 gene cluster found in a pandemic clone of Vibrio parahaemolyticus. The genes for this V. cholerae TTSS system appear to be present in many clinical and environmental non-O1, non-O139 strains, including at least one clone that is globally distributed. We hypothesize that the TTSS present in some pathogenic strains of non-O1, non-O139 V. cholerae may be involved in the virulence and environmental fitness of these strains.     

Genomic diversity of 2010 Haitian cholera outbreak strains

The millions of deaths from cholera during the past 200 y, coupled with the morbidity and mortality of cholera in Haiti since October 2010, are grim reminders that Vibrio cholerae, the etiologic agent of cholera, remains a scourge. We report the isolation of both V. cholerae O1 and non-O1/O139 early in the Haiti cholera epidemic from samples collected from victims in 18 towns across eight Arrondissements of Haiti. The results showed two distinct populations of V. cholerae coexisted in Haiti early in the epidemic. As non-O1/O139 V. cholerae was the sole pathogen isolated from 21% of the clinical specimens, its role in this epidemic, either alone or in concert with V. cholerae O1, cannot be dismissed. A genomic approach was used to examine similarities and differences among the Haitian V. cholerae O1 and V. cholerae non-O1/O139 strains. A total of 47 V. cholerae O1 and 29 V. cholerae non-O1/O139 isolates from patients and the environment were sequenced. Comparative genome analyses of the 76 genomes and eight reference strains of V. cholerae isolated in concurrent epidemics outside Haiti and 27 V. cholerae genomes available in the public database demonstrated substantial diversity of V. cholerae and ongoing flux within its genome.     

非O157产志贺毒素大肠杆菌分离株的多位点序列分型研究

目的 对国内部分地区非O157产志贺毒素大肠杆菌分离株进行分子分型分析,了解菌株间的遗传进化关系。 方法 根据mlst.ucc.ie数据库提供的大肠杆菌多位点序列分型（Multilocus Sequence Typing, MLST）方案,对来自我国河南、黑龙江2省的29株产志贺毒素大肠杆菌分离株的7个管家基因进行PCR扩增并测序,通过序列比对确定其等位基因谱及菌株序列型（Sequence type, ST）,使用MEGA、eBURST生物信息软件分析不同ST序列群及菌株间的进化关系。结果 29株非O157产志贺毒素大肠杆菌呈现较大的遗传多态性,可分为13个ST型别,其中ST155为优势型别（34.48%）。同时研究发现2个新等位基因型（fumC376、recA214）和3个新序列型（ST2460、ST2467、ST2468）。结论 29株非O157产志贺毒素大肠杆菌菌株之间具有分子多态性,与国际流行菌株具有一定的亲缘关系。加强我国对这一类菌株的检测和监测具有重要的公共卫生意义。     

Increasing drug resistance in Vibrio cholerae O1 and non-O1 strains isolated from diarrheal cases in Japan

Drug resistance trends were investigated for 271 Vibrio cholerae O1 (V.c O1) and 401 V. cholerae non-O1 (V.c non-O1) strains isolated from mainly imported diarrheal cases during 1981-2001 in Japan. The results of drug resistance test using 8 drugs (CP, TC, SM, KM, ABPC, ST, NA, and NFLX) showed that 34.7% of the V. c O1 strains and 15.7% of V.c non-O1 strains were multi-drug or mono-drug resistant. The incidence of drug resistant strains has increased since 1991, and it has been remarkable in V.c O1 strains that increased from 1.2% in 1981-1985 to 70.8% in 1996-2001. The drug resistance patterns of the resistant strains classified into 6 types in V.c O1 and 21 types in V.c non-O1. The prevalent patterns recognized were SM (75.5%), CP.TC.SM.ST (10.6%) and CP.SM.ST (8.5%) in V.c O1, and SM (25.4%) and ABPC (25.4%) in V.c non-O1. Ten V.c O1 strains (3.7%) and 10 V.c non-O1 strains (2.5%) were multi-drug resistant including TC. Among those, 13 strains were isolated from travelers who returned to Japan from Thailand. One V.c O1 strain (0.4%) and 6 V.c non-O1 strains (1.5%) were NA high-resistant and fluoroquinolones low-sensitive. Among those, 4 strains were isolated from travelers who returned to Japan from India.     

Genetic diversity and virulence potential of environmental Vibrio cholerae population in a cholera-endemic area

To understand the evolutionary events and possible selection mechanisms involved in the emergence of pathogenic Vibrio cholerae, we analyzed diverse strains of V. cholerae isolated from environmental waters in Bangladesh by direct enrichment in the intestines of adult rabbits and by conventional laboratory culture. Strains isolated by conventional culture were mostly (99.2%) negative for the major virulence gene clusters encoding toxin-coregulated pilus (TCP) and cholera toxin (CT) and were nonpathogenic in animal models. In contrast, all strains selected in rabbits were competent for colonizing infant mice, and 56.8% of these strains carried genes encoding TCP alone or both TCP and CT. Ribotypes of toxigenic O1 and O139 strains from the environment were similar to pandemic strains, whereas ribotypes of non-O1 non-O139 strains and TCP(-) nontoxigenic O1 strains diverged widely from the seventh pandemic O1 and the O139 strains. Results of this study suggest that (i) the environmental V. cholerae population in a cholera-endemic area is highly heterogeneous, (ii) selection in the mammalian intestine can cause enrichment of environmental strains with virulence potential, (iii) pathogenicity of V. cholerae involves more virulence genes than currently appreciated, and (iv) most environmental V. cholerae strains are unlikely to attain a pandemic potential by acquisition of TCP and CT genes alone. Because most of the recorded cholera pandemics originated in the Ganges Delta region, this ecological setting presumably favors extensive genetic exchange among V. cholerae strains and thus promotes the rare, multiple-gene transfer events needed to assemble the critical combination of genes required for pandemic spread.     

Seasonality and antimicrobial resistance pattern of Vibrio cholerae in a tertiary care hospital of North India

We retrospectively analysed the seasonal distribution of cholera and the antimicrobial resistance pattern of Vibrio cholerae isolates over a 5-year period from January 1998 to December 2002. Of 3213 stool specimens processed from 3213 patients with acute watery diarrhoea during this period, 431 samples (13.4%) were found positive for V. cholerae. There were 423 V. cholerae O1 biotype E1 Tor, 2 V. cholerae O139 and six isolates of non-O1 non-O139. The highest number of cholera cases occurred in May-June followed by July-August. Cases started appearing in April for all years except in the year 2002 when three cases occurred in the first week of March. A large number (90.25% strains) were resistant to at least one antibiotic.     

Distribution of virulence markers in clinical and environmental Vibrio cholerae non-O1/non-O139 strains isolated in Brazil from 1991 to 2000

One hundred seventy nine Vibrio cholerae non-O1/non-O139 strains from clinical and different environmental sources isolated in Brazil from 1991 to 2000 were serogrouped and screened for the presence of four different virulence factors. The Random Amplification of Polymorphic DNA (RAPD) technique was used to evaluate the genetic relatedness among strains. Fifty-four different serogroups were identified and V. cholerae O26 was the most common (7.8%). PCR analysis for three genes (ctxA, zot, ace) located of the CTX genetic element and one gene (tcpA) located on the VPI pathogenicity island showed that 27 strains harbored one or more of these genes. Eight (4.5%) strains possessed the complete set of CTX element genes and all but one of these belonged to the O26 serogroup suggesting that V. cholerae O26 has the potential to be an epidemic strain. The RAPD profiles revealed a wide variability among strains and no genetic correlation was observed.     

两株布鲁菌MLST新序列型(ST)的鉴定

目的 对北京地区分离自布病患者血液的2株布鲁菌进行多位点序列分型(Multilocus sequence typing,MLST)及其遗传学特征进行研究,为布病的预防和控制提供科学依据。方法 应用布鲁菌属特异性PCR(BCSP31-PCR)和种/型特异性PCR(AMOS-PCR)对分离的2株布鲁菌进行鉴定,采用MLST技术对2株布鲁菌分离株的19个管家基因、1个外膜蛋白基因及1个基因间区的序列进行测定,与MLST数据库中的等位基因序列进行比对,确定菌株的等位基因谱及9位点和21位点序列型(sequence type,ST)。采用聚类分析法研究2株菌ST型与各种布鲁菌已知ST型的遗传进化关系。结果 2株分离株经BCSP31-PCR鉴定为布鲁菌属细菌,AMOS-PCR鉴定为非羊种、非牛种1、2、4型、非猪种1型、非绵羊附睾种布鲁菌;MLST分析显示,2株菌的等位基因编号(gap/aroA/glk/dnaK/gyrB/trpE/cobQ/int-hyp/omp25/prpE/caiA/csdB/soxA/leuA/ mviM/fumC/fbaA/ddlA/putA/mutL/acnA)分别为2、1、2、2、1、3、1、4、1、13、2、2、2、2、1、1、3、7、6、2、3和2、1、2、2、1、3、1、4、29、13、2、2、2、2、1、1、3、7、6、2、3,比对MLST数据库,结果显示这2株细菌的等位基因编号组合未见报道,为新ST型。聚类分析显示这2种新的ST型与牛种布鲁菌ST型处于同一分支。结论 北京地区分离的2株布鲁菌为新ST型牛种布鲁菌,已被MLST数据库确认,分别命名为ST74和ST75(9位点序列型)与ST108和ST109(21位点序列型)。与同属牛种布鲁菌的ST型遗传关系最近,该结果为北京地区布病的预防和控制提供了科学依据。