目的应用T-RFLP分析胆固醇结石中微生物群落结构。方法应用末端标记限制性片段长度多态性（Terminal Restriction Fragment Length Polymorphism，T-RFLP）和克隆文库分析，以微生物群落16S rRNA基因（16S rDNA）为目标，对8例常规细菌培养阴性的纯胆固醇患者胆囊结石和胆汁中的微生物群落结构进行解析和比较。结果发现8例纯胆固醇结石患者胆汁中未找到细菌存在的证据，结石中细菌16S rDNA阳性率为37.5%（3/8），细菌16S rDNA片段测序表明细菌群落与肠杆菌科和微球菌科的微生物有较高的相似性，同时细菌群落中检出了大量的未培养微生物（Uncultured bacterium clone）。结论运用T-RFLP方法分析16S rDNA克隆片段能够有效评估纯胆固醇结石中的细菌群落结构的多样性。

Objective To analysis of microbial community structure in cholesterol calculus by T-RFLP (Terminal Restriction Fragment Length Polymorphism). Methods Small subunit rRNA gene (16S rDNA) was retrieved from 8 patients'' gallstone and bile with cholesterol calculus. The microbial community structure of these cholesterol calculus patients'' gallstone and bile were investigated by T-RFLP and clone libraries approaches. Results There was no bacterium found in bile from 8 patients with cholesterol calculus. The positive rate of bacterial 16S rDNA in stone was 37.5%(3/8). Bacterial 16S rDNA sequencing fragments showed that microbial community and Enterobacteriaceae and Micrococcaceae microorganisms had high similarity. Meanwhile, there were detected a number of uncultured bacterium from microbial community. Conclusion Analysis of 16S rDNA cloned fragment could effectively evaluate the diversity of bacterial community structure by T-RFLP method in cholesterol stone.

目的：检测肠球菌的主要毒力基因，并测定其生物膜形成情况。方法收集血液来源标本中的粪肠球菌28株，屎肠球菌54株，采用多重PCR检测肠球菌的5种主要毒力基因：asa1、esp、hyl、cylA及gelE；利用微孔板法检测生物膜的形成。结果粪肠球菌中asa1、esp、cylA、gelE的同时检出率为50％，且28株菌均检测到了至少一种毒力基因，仅在粪肠球菌中检测到了asa1、cylA和gelE基因；屎肠球菌对esp的检出率分别为50％，hyl和esp的同时检出率为22．2％，hyl基因仅在屎肠球菌中存在，18．5％的屎肠球菌没有检测到5种毒力基因中的任何一种。粪肠球菌和屎肠球菌形成生物膜阳性菌株的比率分别为85．7％和63．0％。结论致血液感染粪肠球菌在主要毒力基因的种类、毒力基因检出率及生物膜形成阳性菌株数均高于屎肠球菌。

Objective To detect the main virulence genes and biofilm formation of Enterococci strains isolated from blood samples .Methods Twenty-eight strains of Enterococcus faecalis ( E.faecalis) and 54 strains of Enterococcus faecium ( E.faecium) were collected from blood samples .Five main virulence genes (asa1, esp, hyl, cylA and gelE) were detected by multiplex PCR.Biofilm formation was investigated by using microtiter dish biofilm formation assay .Results All E.faecalis strains were positive for at least one kind of virulence genes , of which 14 strains were concurrently positive for asa1, esp, cylA and gelE.asa1, cylA and gelE were only detected in E.faecalis strains, while hyl gene only existed in E.faecium strains. Twenty-seven strains of E.faecium were esp positive, of which 12 strains were both hyl and esp positive. None of the 5 virulence genes were identified in 10 strains of E.faecium.85.7% of E.faecalis strains and 63.0%of E.faecium strains could form biofilm.Conclusion Compared w

目的：了解2009～2013年该院临床分离粪肠球菌和屎肠球菌对各类抗菌药物的耐药性。方法临床分离菌采用最低抑菌浓度法进行细菌的药敏试验，结果按临床实验室标准化协会2013年版的标准进行判断。结果共分离得到非重复粪肠球菌819株，屎肠球菌726株，这两种菌株对利奈唑胺和万古霉素仍很敏感，耐药率均低于1．5％。粪肠球菌和屎肠球菌对万古霉素的耐药率分别为0．1％和1．5％。粪肠球菌对青霉素、氨苄西林和呋喃妥因的耐药率较低，分别为4．3％、1．6％和1．6％；屎肠球菌对青霉素和氨苄西林的耐药率都在87．0％以上，明显高于粪肠球菌。结论该院的肠球菌感染中以粪肠球菌为主，屎肠球菌感染次之，二者的耐药性差异明显，持续监测其耐药情况有助于指导临床合理用药。

Objective To investigate the antimicrobial resistance of E.faecalis and E.faecium isolated from the Hospital from 2009 to 2013.Methods Minimum inhibitory concentration method was used for antimicrobial susceptibility test of the clinical iso-lates.Results were judged according to Clinical and Laboratory Standards Institute′s 2013 version of criterias.Results A total of 819 nonduplicated E.Faecalis and 726 nonduplicate E.faecium isolates were collected.The strains were still highly susceptible to linezolid and vancomycin,to which the drug-resistance rates were both less than 1.5%.The prevalence of vancomycin-resistant E. faecalis and E.faecium was 0.1% and 1.5%,respectively.The ratio of E.faecalis resistant to ampicillin,penicilin and nitrofuran-toin was 4.3%,1.6% and 1.6%,respectively.More than 87% E.faecium strains were resistant to ampicillin and penicillin,and the E.faecium strains were more resistant to most drugs tested than E.faecalis .Conclusion There was an obvious d

肺隐球菌病是由新型隐球菌感染引起的肺深部真菌病。若肺隐球菌病得不到及时诊断治疗,可致全身感染,严重者可引起隐球菌脑膜炎。其好发于艾滋病患者,然而在免疫力正常的个体中报道也逐渐增多。本文回顾性分析1例原发性肺隐球菌病患者的诊断和治疗过程,总结治疗经验,并结合相关文献复习肺隐球菌病的临床表现、诊断与治疗要点,以提高临床医师对肺隐球菌病的认识。

Pulmonarycryptococcosis,causedbycryptococcusneoformansinfectiondeepinlungs,cancausesystemic infection or even cryptococcal meningitis if not diagnosed or treated timely. It often appears in AIDS patients,but increasingly in individuals with normal immunity. This paper analyzes retrospectively the diagnosis and treatment process of pulmonary cryptococ-cosis,summarizes the experience of treatment and reviews the literatures on its clinical manifestations,the key points for its diag-nosis and treatment,to improve clinicians′awareness about pulmonary cryptococcosis.

了解肠球菌的临床分离率及其对常用抗生素的耐药性,指导临床合理使用抗生素。方法:用Vitek 2 Compact法进行细菌鉴定及常规药敏试验。结果:临床分离到的342株肠球菌中,粪肠球菌有140例,阳性率40.9%,屎肠球菌有186例,阳性率54.4%,其它肠球菌有16例,阳性率4.7%。342株肠球菌主要分离自尿液254例,阳性率74.3%;血液28例,阳性率8.2%;脑脊液21例,阳性率6.1%;胆汁15例,阳性率5.3%。屎肠球菌对青霉素、氨苄西林、环丙沙星、左氧氟沙星、红霉素和四环素的耐药率已接近或达到90%,而粪肠球菌对青霉素和氨苄西林仍保持100%敏感性。未发现肠球菌对万古霉素、利奈唑胺和替加环素耐药。结论:革兰阳性球菌中肠球菌是引起医院感染的重要病原菌之一,肠球菌引起的感染性疾病的治疗一定要以药敏结果为指导合理选用抗生素。

Objective:To understand the distribution and drug resistance of commonly used antibiotics of enterococcus, in order to provide the basis for clinical rational drug use. Methods:Vitek 2 Compact method was used for routine bacteria identification and drug susceptibility test. Results: Among clinical isolated 342 strains of enterococcus, 140 cases were 40.9% in enterococcus, excrement enterococcus 186 rate of 54.4%;other enterococcus 16 cases were 4.7%. 342 strains of enterococcus were mainly isolated from urine of 254 cases (74.3%), 28 cases (8.2%) and blood cerebrospinal fluid 21 cases (6.1%), 15 cases (5.3%) of bile. Excrement enterococcus to penicillin, ampicillin, ciprofloxacin, levofloxacin, erythromycin and tetracycline resistance was close to or reached 90%, and dung bowel aureus to penicillin and ampicillin remained 100%sensitivity. There were no resistance to enterococcus among vancomycin, rina thiazole amine and Tigecycline. Conclusion: Enterococcus was one of the imp