分离自腹泻成人志贺菌的血清分群及耐药性分析

目的 调查志贺菌在成人腹泻患者中的主要血清群和耐药性,为临床治疗和预防提供依据.方法 对2008年1月至2012年10月浙江萧山医院临床分离自腹泻成人志贺菌菌株,进行生化鉴定、血清学分群和抗生素敏感性试验,所有数据用WHONET 5.6软件进行回顾性分析.结果 共检出125株志贺菌,其中66(52.8％)株为宋内志贺菌,58(46.4％)株为福氏志贺菌;氨苄西林耐药率最高达95.9％,复方新诺明耐药率达79.8％,头孢噻肟为48.6％,未检出头孢哌酮/舒巴坦、哌拉西林/他唑巴坦、亚胺培南和美罗培南耐药株,福氏志贺菌对环丙沙星和左旋氧氟沙星耐药率为87.0％和57.1％,而宋内志贺菌未检出氟喹诺酮类耐药株;福氏志贺菌和宋内志贺菌耐药率在哌拉西林、头孢他啶、头孢噻肟、头孢吡肟、复方新诺明和氟喹诺酮间差异有统计学意义(P＜0.01).结论 萧山地区成人志贺菌感染以宋内和福氏两种血清群为主,青霉素类和复方新诺明耐药率太高已不适合临床使用,含酶抑制剂抗生素是治疗志贺菌感染的首选药物.     

苏州地区儿童感染志贺菌的耐药监测

目的调查苏州地区2000年至2006年儿童感染志贺菌的耐药情况,了解其耐药趋势,以指导临床合理用药。方法对2000年至2006年苏州大学附属儿童医院临床标本中分离的589株志贺菌（福氏志贺菌470株,宋内志贺菌119株）采用K-B法对氨苄西林、哌拉西林、头孢哌酮、头孢曲松、环丙沙星、SMZ＋TMP、亚胺培南进行药敏试验。结果于7年中470株福氏志贺菌的耐药率：氨苄西林：始终在90．0％以上;哌拉西林：从22．6％逐渐上升到63．2％;头孢哌酮：1．1％～56．1％;头孢曲松：2．2％～43．9％;环丙沙星：一直在10．0％左右;SMZ＋TMP：在70．0％～96．0％波动;对亚胺培南均敏感。119株宋内志贺菌的耐药率：氨苄西林：从7．1％逐渐上升到82．8％;哌拉西林：0～79．3％;头孢哌酮：0～55．2％;头孢曲松：0～51．7％;SMZ＋TMP：在62．5％～100．0％波动;对环丙沙星和亚胺培南均敏感。结论本地区儿童感染福氏志贺菌对氨苄西林严重耐药,同时伴有对SMZ＋TMP的高耐药率,对哌拉西林和3代头孢菌素的耐药率呈逐年上升趋势。宋内志贺菌的耐药率在2004年之前（除对复方新诺明外）,远低于福氏志贺菌,但在2005年其对氨苄西林和3代头孢菌素的耐药率突然上升,大有赶超福氏志贺菌之势。因此苏州地区儿童感染志贺菌的耐药情况不容乐观,对儿童细菌性痢疾的治疗将会面临困境。     

临床分离志贺菌中CRISPR/Cas系统的分布及其与毒力基因的关系

【目的】了解临床分离志贺菌中CRISPR/Cas系统的分布特征并分析其与毒力基因的关系。【方法】以聚合酶链式反应(PCR)方法,采用10对引物分别对57株临床分离志贺菌中CRISPR1、cas2-cas1、cas6e-cas5、cas7、cse2、cse1-cas3基因和毒力基因ipaH、ial、ipaBCD、virA进行检测。对CRISPR1的PCR结果进行测序,并用CRISPR finder在线软件对CRISPR1基因座进行分析。通过卡方检验初步分析CRISPR/Cas系统与毒力基因的关系。【结果】测序结果显示,CRISPR1基因座中间隔序列数目较少且在不同菌株间一致性较高;57株志贺菌中,84.2% (48/57)的志贺菌中可检测到CRISPR/Cas系统,其中68.8% (33/48)的志贺菌中cas6e-cas5基因或(和) cse2基因中发现插入序列;毒力基因ipaH、ial、virA、ipaBCD的检出率依次为100%、100%、98.2%和87.7%;毒力基因ipaBCD的阳性率与活性CRISPR/Cas系统的分布无关(P>0.05)。【结论】CRISPR/Cas系统广泛存在于临床分离志贺菌中;部分cas基因中有插入序列;并未发现志贺菌中活性CRISPR/Cas系统与毒力基因的分布有关。     

183株志贺菌的菌群分布及药敏分析

目的了解本地区细菌性痢疾菌群分布及药敏情况。方法对183株志贺菌进行菌群分型及药敏分析。结果183株志贺菌有福氏志贺菌168株（91．8％）;宋内志贺菌9株（4．9％）;痢疾志贺菌4株（2．2％）;鲍氏志贺菌2株（1．1％）。药敏试验显示志贺菌对四环素、氨苄西林、复方新诺明耐药严重;对头孢他啶、阿米卡星、诺氟沙星比较敏感。结论本地区志贺菌主要是福氏志贺菌Ⅱa,应根据药敏试验结果选择合适抗菌药物。     

产志贺毒素大肠埃希菌的分子生物学鉴定和耐药性分析

为了解产志贺毒素大肠埃希菌 (Shigatoxin producingEscherichiacoli ,STEC)stx1,stx2 ,eaeA ,hlyA 4种毒力基因的分布情况 ,以及分离株对 18种抗生素的敏感性 ,采用多重PCR(multiplexPCR ,mPCR)法对分离株进行毒力基因的分子生物学鉴定 ;用WHO推荐的K B法对分离株进行抗生素的敏感性测定。产志贺毒素的大肠埃希菌共有 4 6株 ,其中 2种毒素均产生的有 2 2株 (4 7.8% ) ;单纯产生stx1的有 16株 (36 .9% ) ,stx2 的有 8株 (17.4 % ) ;4种毒力基因均存在的有 19株 (4 1.3% ) ,血清型为O15 7∶H7,而非O15 7∶H7血清型的菌株 (2 3/46 )中 ,4种毒力基因同时存在的仅有 3株 (6 .6 % ) ,但有 13株 (5 6 .9% )hlyA基因阳性。全部STEC对复方新诺明耐药 ,对链霉素耐药率为 2 8.3% ,氨苄西林为 30 .4 % ,红霉素为 6 9.6 % ,而且有 5株对至少 4种以上抗生素多重耐药 ,耐药谱为复方新诺明 链霉素 红霉素 氨苄西林。非O15 7型STEC耐药菌次为 12 2 ,而O15 7型为 6 3。可见 ,mPCR法可以快速检测STEC特征性毒力基因 ,以判定其致病性能。非O15 7型STEC对抗生素较易形成耐药性。     

不同地区宋内志贺菌耐药性及脉冲场凝胶电泳分型分析

摘要:【目的】通过对不同地区的宋内志贺菌株进行药物敏感性检测、耐药基因的扩增以及基因分型,了解不同地区宋内志贺菌的耐药情况与流行趋势。【方法】使用微量肉汤稀释法测定了54 株宋内志贺菌对21种药物的敏感性,用PCR方法扩增相关耐药基因,利用脉冲场凝胶电泳(Pulsed Field Gel Electrophoresis,PFGE)技术进行宋内志贺菌的基因分型,最后采用BioNumerics分析软件对所有菌株进行聚类,分析其相似度。【结果】实验菌株对于甲氧苄氨嘧啶/磺胺甲噁唑、四环素、替卡西林/棒酸、氨苄西林、庆大霉素5种抗生素普遍耐药,对亚胺培南、头孢吡肟、左氟沙星、诺氟沙星、阿米卡星5种抗生素全部表现为敏感。共检测出包括blaTEM,blaCTX以及整合子在内的7 种不同的耐药基因。全部实验菌株可分为26个不同的PFGE带型,分型后表现出较高的基因同源性。宋内志贺菌的耐药性、携带基因与带型具有一定地域相关性。【结 论】目前各地区流行的宋内志贺菌株对甲氧苄氨嘧啶/磺胺甲噁唑与四环素已经普遍耐药,不同地区出现相同聚类分型的菌株表明存在跨区域流行的宋内志贺菌群。因此,加强对不同地区宋内志贺菌的监控对减少多重耐药菌株的产生具有重要意义。     

志贺氏菌福氏2a 301株ipaH4.5突变株的构建及生物学功能分析

利用λ-Red重组系统对福氏2a志贺氏菌301株ipaH4.5基因进行缺失突变,构建了福氏2a志贺氏菌301株ipaH4.5基因缺失突变株?ipaH4.5,利用低拷贝质粒构建ipaH4.5缺失突变株的回复突变株△ipaH4.5HF。PCR方法证实了ipaH4.5基因的缺失和回复。对野生株、突变株和回复突变株的生长代谢及细胞侵袭能力进行比较;ELISA方法检测3株菌侵袭鼠J774巨噬细胞后培养上清中炎性因子的水平。生长代谢实验表明缺失和回复ipaH4.5不影响志贺氏菌的生长速度,侵袭实验表明缺失和回复ipaH4.5也不影响志贺氏菌对HeLa细胞和鼠J774巨噬细胞的侵袭能力,表明ipaH4.5基因与志贺氏菌的生长代谢和侵袭能力无关;鼠J774巨噬细胞培养上清中细胞因子水平的改变提示该基因在志贺氏菌侵入细胞后抑制宿主细胞炎症反应。     

志贺菌的质粒图谱分析

目的：分析志贺菌的质粒图谱及其与细菌药物敏感性的相关性。方法：从菌痢患者粪便标本中分离6株福氏志贺菌和4株宋内志贺菌,分别对其质粒图谱及药物敏感性进行分析。结果：不同菌株的质粒图谱具有明显的差异,但福氏志贺菌的5株以及宋内志贺菌的3株具有分子量23Kb的质粒带。各菌株的质粒图谱与其对头孢三嗪,头孢唑啉,环丙沙星,诺氟沙星,氯霉素的耐药特性无明显相关性。结论：获自患者的福氏志贺菌和宋内志贺菌具有不同的质粒图谱以及抗菌药物敏感性,提示在我市引起菌痢的志贺菌具有不同的来源。     

福氏志贺菌CTX-M型产超广谱β-内酰胺酶的检测及耐药性

目的 产超广谱β-内酰胺酶( extended-spectrum β-lactamases,ESBLs)福氏志贺菌的耐药性及其基因型.方法 K-B法检测福氏志贺菌对抗菌药物的敏感性,改良三维试验检测菌株是否产生ESBLs,ESBLs的基因型通过PCR扩增、DNA测序的方法进行.结果 148株福氏志贺菌检测出6株产ESBLs,其基因型为CTX-M-3和CTX-M-14;产ESBLs福氏志贺菌对氨苄西林、头孢噻肟、复方新诺明、四环素均耐药,对头孢西丁、亚胺培南、环丙沙星敏感.结论 应加强对福氏志贺菌的耐药性监测,合理使用抗生素,防止产ESBLs福氏志贺菌的流行和传播.     

210株肠道致病菌的分类及耐药性分析

目的：探讨本地区肠道致病菌的分类及主要致病菌的耐药性,给临床治疗腹泻疾病提供指导。方法：药敏试验采用K-B法,用WHONET4软件进行数据分析。结果：210株肠道致病菌中,志贺菌属占70％,沙门菌属占3％,致病性大肠埃希菌占4．8％,副溶血弧菌占3．8％,阴沟肠杆菌占3．3％,鼠伤寒沙门菌占2．4％,霍乱弧菌占1．9％,其他占10．8％等;在志贺菌属中,福氏占87％,宋内占13％。福氏和宋内志贺菌对三代头孢和IMP较敏感,对复方新诺明耐药率较高。结论：福氏志贺菌是肠道的主要致病菌;临床怀疑菌痢时,应首选三代头孢、IMP。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.