肺炎克雷伯菌第1类整合子的鉴定和耐药性分析

目的 了解来源于不同临床标本的肺炎克雷伯菌的耐药性、携带第1类耐药整合子的特征及基因盒的种类.方法 使用常规方法分离肺炎克雷伯菌;运用法国梅里埃VITEK细菌分析系统进行鉴定;应用纸片扩散法对15种抗生素进行耐药性监测和分析;应用PCR法扩增第1类整合子;对PCR产物纯化、测序,并对结果进行分析.结果 来源于不同临床标本的64株肺炎克雷伯菌100%耐药,32株(50%)表现多重耐药,9株耐4种抗生素,耐药谱为氨苄西林-链霉素-复方新诺明-四环素;20株(31.2%)为第1类整合子阳性株,其中17株整合子大小为1.0 kb,2株为1.6 kb,1株含有1.0 kb和1.6 kb两个整合子;PCR产物测序显示1.0 kb整合子携带pse-1-aadA1基因盒,1.6 kb整合子插入了pse-1-aadA2,pse-1-aacA1基因盒,双重整合子株插入了pse-1-aadA1和pse-1-aacA1基因盒.结论 第1类整合子存在于各种临床标本分离的肺炎克雷伯菌中,且与多重耐药性相关,主要决定着对β内酰胺类和氨基糖苷类抗生素的耐药性.     

多重耐药鲍曼不动杆菌携带金属酶与整合酶基因特征及同源性分析

目的了解吉林地区多重耐药鲍曼不动杆菌(Multidrug-resistant Acinetobacter baumanni,MDRAB)金属酶与整合酶基因的类型,并对菌株进行同源性分析.方法收集吉林地区2所三级甲等医院临床标本分离鉴定的鲍曼不动杆菌38株.多重耐药株携带金属酶的初筛实验-双纸片协同试验:金属酶与整合酶基因的检测-聚合酶链反应(PCR),并对其进行同源性分析.结果在所有MDRAB菌株亚胺培南-EDTA协同试验中,15株(39.4%)阳性,16株(42.1%)携带整合酶基因Ⅰ,2株(5.2%)携带整合酶基因Ⅱ,2株(5.2%)携带blaVIM型金属酶基因,1株(2.6%)携带blaNDM-1型金属酶基因,未检测出金属酶基因blaIMP,blaSIM-1和整合酶基因Ⅲ.结论该地区临床分离的MDRAB耐药率高,可能与其携带金属酶基因和整合酶基因有关,A型MDRAB为医院感染主要流行株.     

肾移植术后感染致病菌的整合子分类及耐药分析

目的:探讨肾移植患者病原菌的整合子基因分类与耐药的相关性,以期控制和预防院内感染,合理使用抗生素。方法:收集我院19例肾移植患者的19株细菌培养结果和药敏试验的资料,用多重PCR方法进行病原菌整合子的基因检测,并用K-B法与M IC法对其进行药敏分析。结果:①19株致病菌在体外药敏实验中都对抗生素产生严重的多重耐药。②产整合酶基因的菌株共12株,占63%;其中革兰阳性球菌3株,占25%;革兰阴性杆菌9株,占75%;其中2株为耐甲氧西林凝固酶阴性葡萄球菌(MRSCON),4株为产超广谱β-内酰胺酶(ESBL)的大肠杆菌,1株为产ESBL的肺炎克雷伯菌,其余为肠球菌1株、肠杆菌3株和洋葱伯克霍尔德菌1株。③产整合酶均为Ⅰ类整合酶,其中检测出耐药基因盒为1 009 bp的5株,1 664 bp 1株。结论:肾移植患者的病原菌都存在严重的多重耐药性并与整合酶基因有密切相关。对肾移植患者病原菌进行整合酶基因与药敏分析是控制和预防病区交叉感染、合理使用抗生素治疗多重耐药细菌,降低病死率的关键因素。     

成都地区60株铜绿假单胞菌中第I类整合子的鉴定及特性分析

采用两种PCR法(整合酶PCR法和长片段整合子PCR法)检测60株成都地区4家医院分离的铜绿假单胞菌的第Ⅰ类整合子,并对扩增产物进行序列分析.对60株铜绿假单胞菌第Ⅰ类整合酶基因的检测,发现有28株扩增结果为阳性(46.7%),所扩增片段的大小为110 bp.测序结果与GenBank中已报道的ⅠntI1基因核酸序列同源性为100%.长片段PCR法扩增Ⅰ类整合子的可变区,60株铜绿假单胞菌中有18株检测到不同大小的Ⅰ类整合子基因盒,占标本总数的30.0%.插入的基因盒大小从300 bp到超过2000 bp不等,18株整合子阳性菌中,有两株菌同时含有2种大小不同的整合子.序列分析结果表明,菌株R03的1900 bp 整合子携带aacA4基因盒,菌株W13的1900 bp整合子中含dfr17和aadA5基因盒.     

肠杆菌科细菌耐药基因NDM和ESBLs的检测及白头翁汤增强其敏感性研究

采用美国临床实验室标准化协会(Clinical and Laboratory Standards Institute, CLSI)推荐的方法，检测了临床分离的44株肠杆菌科细菌产超广谱β-内酰胺酶(Extended-Spectrum Beta-Lactamases, ESBLs)的情况.利用PCR对临床分离菌进行新德里金属β-内酰胺酶(New Delhi metallo-β-lactamase, NDM)以及ESBLs耐药基因的检测.用微量肉汤稀释法测定了分离菌对阿莫西林等14种抗菌药的敏感性，并统计其耐药率.结果表明，临床分离的44株肠杆菌科细菌中，有31株产超广谱β-内酰胺酶，检出率为70.5%.耐药基因检测结果显示，耐药基因NDM-1携带率为13.6%.ESBLs耐药基因TEM,SHV,CTX-M,OXA的携带率分别为100%,43.2%,45.5%,6.8%.同时发现，有6株菌株同时检出了含NDM-1和ESBLs的基因，有9株菌株同时检出含有2种ESBLs基因，3株同时检出含有3种ESBLs基因.敏感性测定结果显示，分离菌大多呈现多重耐药，分离菌除对替加环素(18.2%)和阿米...     

规模化鸡场粪样和苍蝇产CMY-2大肠杆菌中耐药基因与毒力基因调查及其共转移研究

为了解全国20家规模化鸡场粪样和苍蝇产CMY-2大肠杆菌中耐药基因与毒力基因的流行现状及其共转移情况,对2013-2015年间分离自粪样和苍蝇中的549株大肠杆菌,采用聚合酶链反应（PCR）方法筛选出blaCMY-2阳性菌株;脉冲场凝胶电泳（PFGE）分析blaCMY-2阳性菌株的遗传进化关系;K-B纸片法检测阳性菌株对13种抗菌药物的耐药性;利用PCR技术检测19种相关耐药基因以及14种毒力基因;接合转移试验和质粒复制子分型研究耐药基因及毒力基因的传播方式。结果,33株大肠杆菌呈CMY-2阳性,阳性率为6.0 %,且均表现为多重耐药;检测到15种耐药基因（blaTEM-1、blaCTX-M-55、blaOXA-1、qnrBDS、aac(6’)-Ib-cr、oqxA、tetA、tetC、sul1、sul2、sul3、rmtB和flor）和4种毒力基因（iutA、traT、fyuA和VagC）;33株携带blaCMY-2基因菌株中有21株接合转移成功,blaCMY-2基因可通过lncA/C或lncI1质粒与耐药基因（blaTEM-1、blaCTX-M-55、tetA或sul2）和（或）毒力基因（traT或VagC）共同转移。本研究阐明了规模化鸡场产CMY-2大肠杆菌携带耐药基因与毒力基因的情况,证实了细菌中质粒介导的耐药与毒力的共转移,为耐药性传播及疾病防控提供了参考。     

耐碳青霉烯酶肺炎克雷伯菌的分离鉴定及生物学特性分析

以临床尿路感染患者尿液为研究对象,通过分离培养及16s rDNA进行肺炎克雷伯菌的分离鉴定;采用Kirby-Bauer纸片法进行药敏实验,并对分离株进行碳青霉烯酶表型筛选;通过PCR检测常见的碳青霉烯酶耐药基因、荚膜血清型和毒力基因分布情况;对分离的肺炎克雷伯菌株进行了生物被膜形成能力及其对小鼠致病力的分析。本研究从采取的86例尿液标本中分离检出11株耐碳青霉烯酶肺炎克雷伯菌,分离率为12.79%。耐碳青霉烯酶基因PCR检测结果显示,blaNDM、blaVIM、blaIMP和blaKPC基因在分离株中均呈现不同程度的分布。耐药性分析发现11株肺炎克雷伯菌对氨苄西林耐药率为90.91%,对头孢噻肟耐药率为63.64%,对链霉素、庆大霉素、氯霉素和诺氟沙星的耐药率较低。耐碳青霉烯类肺炎克雷伯菌荚膜分型结果显示,分离的11株细菌中10株均为强毒力型菌株（90.9%）,其中K57血清型4株（36.4%）,K1血清型1株（9.1%）,K2血清型1株（9.1%）,K5血清型1株（9.1%）,K20血清型3株（27.3%）,提示该批分离株具有较强的致病力。此外,毒力因子分布结果显示,其毒力因子rmpA（54.5%）、Aerobactin F（54.5%）在菌株中分布较为广泛。生物被膜形成能力检测及小鼠致病性试验结果显示,9株分离株均有较强的生物被膜形成能力,菌株致病力可能与荚膜血清型及生物被膜形成能力相关。综上所述,临床分离的致尿路感染病原肺炎克雷伯菌呈现多重耐药特征,强毒力菌株以K57荚膜型为主,K1、K2均有分布,提示对尿路感染病原应加强耐药监测,合理使用抗菌药物,有效防控多重耐药和强毒力菌株的感染与流行。     

重症监护病房多重耐药铜绿假单胞菌Ⅰ型整合子的检测及鉴定

 整合子是介导细胞多重耐药的重要机制之一,鉴定了2008年某医院重症监护病房分离的23株多重耐药铜绿假单胞菌1型整合子,并应用脉冲场电泳分析其同源性。1型整合子的阳性率达成60.9%。3种1型整合子基因盒被鉴定,其中整合子blaOXA 10 acc6 Ⅱ cmlA8为首次发现报道。基因盒主要编码氨基糖苷类耐药基因,包括 aacA4, aadA2, aadB, aac6 Ⅱ。脉冲电泳结果表明,23株多重耐药铜绿假单胞菌分为5个基因型,A 型(n=5)、B型 (n=6)、C型 (n=4)、D型(n=2)和E型(n=2)。研究表明编码1型整合子是多重耐药铜绿单胞菌较为普遍的特征,且1型整合子与多重耐药表型存相关。研究结果同时也表明防止多重耐药铜绿单胞菌交叉感染仍然是ICU的一项挑战性工作。     

养殖环境病原菌以及超级细菌气溶胶的发生与传播的监测

为了观察和评估养殖环境耐药细菌气溶胶的形成与扩散,采用Andersen-6级空气样品收集器在养鸡场鸡舍环境舍内、外(到下风方向800 m)收集空气样品;与此同时,采集鸡只粪便样品。对样品分离的大肠杆菌(E.coli)进行药物敏感性测试。结果显示,各个地点分离的E.coli对利福平和青霉素完全耐药,但他们对妥布霉素、庆大霉素敏感;有不同数量的菌株对氟哌酸、链霉素、头孢哌酮、氯霉素、复合磺胺和四环素有不同程度的耐药性。采用多重PCR方法,分别对从鸡、猪、牛舍(共21个)及其环境中分离到的480株大肠杆菌,进行了5种毒素基因即STa,STb,LTa,Stx1和Stx2/Stx2e的检测。结果表明,分离株都携带有一定数量的毒素基因,很多携带2种或2种以上。通过对11个动物舍(鸡、猪、牛、兔)空气和粪便样品分离的426株肠球菌对四环素类(TetM)、氨基糖甙类抗生素及万古霉素VanA和VanB主要耐药基因的检测表明,14.55%的细菌对β-内酰胺酶耐药;分离株都存在不同程度的对四环素类抗生素的耐药性;共检测出31株肠球菌携带vanA和vanB耐药基因;绝大多数肠球菌携带氨基糖苷修饰酶(AME)基因的一种或几种。     

产CTX-M型超广谱β-内酰胺酶肺炎克雷伯菌耐药性分析

目的了解吉林市区肺炎克雷伯菌CTX-M型超广谱β-内酰胺酶（ESBLs）的基因型分类及耐药情况,并对其分子流行病学特征进行分析.方法采用PCR扩增法检测118株肺炎克雷伯菌产CTX-M型ESBLs基因型情况,调查其阳性株对14种常见抗菌药物的耐药情况,对产CTX-M型肺炎克雷伯菌进行AP-PCR同源性分析,并绘制基因分析图谱.结果 118株肺炎克雷伯菌中共扩增出CTX-M型菌株25株,占21.19%;其中CTX-M-1组13株,占11.02%;CTX-M-9组16株,占13.56%;同时含CTX-M-1组和CTX-M-9组菌株4株,占3.39%;扩增未发现CTX-M-2组、CTX-M-8组和CTX-M-25组阳性株.25株产CTX-M型超广谱β-内酰胺酶均为多重耐药菌（耐3种以上抗菌药物）,对红霉素耐药率高达100%;其次耐药率依次为头孢噻肟92%,氨苄西林92%,四环素88%;对亚胺培南耐药率较低,对美罗培南均敏感.25株CTX-M型阳性菌株做随机扩增多态性分析,共发现13种RAPD.结论吉林市区产CTX-M型超广谱β-内酰胺酶菌株均为多重耐药,且对红霉素、头孢噻肟和氨苄西林类耐药率均很高.CTX-M酶在一定程度上存在克隆传播.     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

An Analysis of Catherine＇s Character——Wuthering Heights

Wuthering Heights, Emily Bronte＇s only novel, was published in December of 1847 under the pseudonym Ellis Bell. The book did not gain immediate success, but it is now thought one of the finest novels in the English language. Catherine is the key character of this masterpiece, because everybody and everything center on her though she had a short life. We can understand this masterpiece better if we know Catherine well.     

Lower Paleozoic oil relationships within Williston Basin, Canada

The Williston Basin is a significant petroleum province, containing oil production zones that include the Middle Cambrian to Lower Ordovician, Upper Ordovician, Middle Devonian, Upper Devonian and Mississippian and within the Jurassic and Cretaceous. The oils of the Williston Basin exhibit a wide range of geochemical characteristics defined as "oil families", although the geochemical signature of the Cambrian Deadwood Formation and Lower Ordovician Winnipeg reservoired oils does not match any "oil family". Despite their close stratigraphic proximity, it is evident that the oils of the Lower Palaeozoic within the Williston Basin are distinct. This suggests the presence of a new "oil family" within the Williston Basin. Diagnostic geochemical signatures occur in the gasoline range chromatograms, within saturate fraction gas chromatograms and biomarker fingerprints. However, some of the established criteria and cross-plots that are currently used to segregate oils into distinct genetic families within the basin do not always meet with success, particularly when applied to the Lower Palaeozoic oils of the Deadwood and Winnipeg Formation.     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.     

Design and analysis of generic LBS application developing platform

Location based services is promising due to its novel working style and contents.A software platform is proposed to provide application programs of typical location based services and support new applications developing efficiently. The analysis shows that this scheme is easy implemented, low cost and adapt to all kinds of mobile nework system.     

Achieving Unusual States of Matter Under High Pressure

正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-     

Exchange-Correlation and Electronic Excitation Energies from Pairing Matrix Fluctuations

We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-     

Density Functional Theory for the Response of Periodic Systems to Electric Fields Based on the Vector Potential Approach

正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that