产ESBLs革兰阴性菌的TEM、SHV、CTX-M基因分型研究

目的:检测本院临床产超广谱β-内酰胺酶(ESBLs)革兰阴性菌的TEM、SHV、CTX-M基因型特征。方法:双纸片法确定产ESBLs的临床分离菌,提取质粒DNA,PCR法扩增ESBLs的TEM、SHV、CTX-M基因片段,对临床分离的12株肺炎克雷伯菌,17株大肠埃希氏菌和16株铜绿假单胞菌,共45株革兰阴性菌进行TEM、SHV、CTX-M基因分型研究。结果:本院临床分离出的45株产ESBLs临床分离菌的质粒中,有36株扩增出TEM基因片段,33株扩增出SHV基因片段,31株扩增出CTX-M基因片段,TEM、SHV、CTX-M基因片段的检出率分别为80%、74%和68%。结论:TEM、SHV、CTX-M已成为本院临床分离产ESBLs菌的主要耐药基因型,有必要进一步加强临床耐药基因的监测,为临床用药和医院感染监控提供参考。     

产CTX-M型超广谱β-内酰胺酶大肠埃希菌的研究

目的 :了解四川大学华西医院产超广谱 β 内酰胺酶 (ESBLs)大肠埃希菌中CTX M基因的检出情况及产CTX M酶大肠埃希菌的耐药性和同源性。方法 :对 2 0 0 1年 1 0月～ 2 0 0 2年 5月我院临床分离的 30株产ESBLs大肠埃希菌采用琼脂平皿对倍稀释法和PCR方法进行耐药表型和 β 内酰胺酶基因型分析 ;对扩增的CTX M基因进行DNA序列分析 ;用脉冲场凝胶电泳 (PFGE)分型技术了解产CTX M酶大肠埃希菌同源性。结果 :1 3株菌携带CTX M基因 ,CTX M酶亚型为CTX M 3和CTX M 1 4。产CTX M酶菌株对第三代头孢菌素耐药率高 ,对头孢噻肟…     

杭州和湖州鲍曼不动杆菌β内酰胺酶和氨基糖苷类修饰酶基因研究

目的了解浙江省杭州和湖州自临床分离的鲍曼不动杆菌中β内酰胺酶(β-lactamases,BLA)基因及氨基糖苷类修饰酶(aminoglycoside-modifying enzymes,AMEs)基因存在状况.方法在2000年7月～2004年2月间从省立同德医院和解放军第98医院各分离20株鲍曼不动杆菌,采用微量稀释法测定其对13种抗菌药物的敏感性,采用聚合酶链反应(PCR)及序列分析的方法分析BLA基因(TEM和SHV)及AMEs基因[aac(3)-Ⅰ、aac(3)- Ⅱ、aac(6′)-Ⅰ和ant(3″)-Ⅰ]类型.结果两地分离株多重耐药严重,但对亚胺培南和美洛培南均无耐药.杭州分离株中TEM、SHV、aac(3)-Ⅰ、aac(3)- Ⅱ、aac(6′)-Ⅰ和ant(3″)-Ⅰ基因阳性率分别为45.0%、0.0%、55.0%、15.0%、35.0%和60.0%,湖州分离株分别为100.0%、30.0%、50.0%、10.0%、55.0%和65.0%.序列分析确认为TEM-1亚型广谱BLA和SHV-12亚型超广谱β内酰胺酶基因.其中98医院HZ40株的TEM-1序列及HZO2株的SHV-12序列均已登录GenBank(GenBank注册号分别为:AY263331和AY259163).结论浙江湖州临床分离的鲍曼不动杆菌中TEM、SHV基因阳性率均高于杭州分离株(Ρ分别＜0.01和＜0.05),但4种AMEs基因阳性率差别不大(Ρ均＞0.05).在鲍曼不动杆菌中发现SHV-12 及TEM-1基因分别为国际及国内首次报道.     

吉林地区大肠埃希菌和肺炎克雷伯菌ESBLs的基因分型

目的对产超广谱β-内酰胺酶(ESBLs)的大肠埃希菌和肺炎克雷伯菌携带的ESBLs耐药基因进行分型,为临床合理应用抗生素提供理论依据。方法收集住院患者标本中分离出的51株大肠埃希菌和32株肺炎克雷伯菌,经PCR对上述菌株所携带的ESBLs耐药基因进行分型。结果产ESBLs大肠埃希菌中耐药质粒编码TEM型、SHV型和非TEM非SHV型超广谱β-内酰胺酶基因的百分率依次为80.4%,7.8%和11.8%;而在肺炎克雷伯菌中的百分率依次为78.1%,71.9%和25.0%。多数产ESBLs肺炎克雷伯菌同时产生一种以上的β-内酰胺酶。结论获得了吉林地区大肠埃希菌和肺炎克雷伯菌ESBLs的不同基因型,ESBLs基因型具有地区性差异。     

肠杆菌科产CTX-M酶细菌耐药性与β-内酰胺酶基因型的关系

目的 :了解我院肠杆菌科产CTX M酶细菌耐药性与 β 内酰胺酶基因型的关系。方法 :对 2 0 0 1年 1 0月～ 2 0 0 2年 5月我院临床分离的 4 0株产ESBLs肠杆菌科细菌琼脂对倍稀释法测定 1 0种抗菌药物的最低抑菌浓度 :用针对SHV、TEM、CTX N基因的特异性引物进行PCR扩增确定 β 内酰胺酶基因型 :对扩增的CTX M基因进行DNA序列分析。结果 :产CTX M酶菌中有 93～ 75 %产 2种或 2种以上 β 内酰胺酶。产CTX M酶菌株多重耐药率为 1 0 0 % ,对头孢噻肟、头孢吡肟、头孢他啶、头孢哌酮 /舒巴坦的耐药率分别为 81 .2 5 %、75 %、31 .2 5…     

产质粒介导AmpC酶和ESBLs细菌的耐药性及β-内酰胺酶基因型研究

目的　了解我院同时产质粒介导AmpC酶和超广谱 β 内酰胺酶 (ESBLs)肺炎克雷伯菌与大肠埃希菌的耐药性及其 β 内酰胺酶的基因型特征。方法　 110株临床分离无重复肺炎克雷伯菌与大肠埃希菌耐药株 ,采用酶提取物三维实验检测AmpC酶 ,美国临床实验室标准委员会 (NCCLS)表型筛选和确认实验检测ESBLs ;琼脂二倍稀释法测定抗生素对同时产AmpC酶和ESBLs菌株的最低抑菌浓度 (MICs) ;等电聚焦实验测定 β 内酰胺酶等电点 (pIs) ;质粒接合实验定位耐药基因 ;PCR通用引物扩增AmpC酶与ESBLs基因及其序列测定以确定其基因亚型。结果　同时产AmpC酶和ESBLs菌株在肺炎克雷伯菌与大肠埃希菌中的检出率分别为 7.7% (5 6 5 )、8.9% (4 4 5 )。该类产酶菌株产 2～ 3种pI5 .4～ 9.0 β 内酰胺酶 ,对第三代头孢菌素、氨曲南、头孢美唑和含酶抑制剂复合制剂的敏感性极低 ,但对亚胺培南均敏感。 9株质粒中均检出AmpC酶基因DHA 1亚型 ,其中 1株同时检出ACT 1亚型 ;ESBLs基因亚型分别为CTX M 14、CTX M 3、CTX M 9,各有 4、3、2株 ;有 3株还携带广谱酶TEM 1基因。结论　我院存在同时产质粒介导DHA 1、ACT 1型AmpC酶和CTX M型ESBLs肺炎克雷伯菌、大肠埃希菌流行株 ,药敏检测结果显示对大多数新型广谱 β 内酰胺类抗生     

肺炎链球菌青霉素结合蛋白基因pbp2x新的变异与青霉素及头孢噻肟耐药

目的 调查研究肺炎链球菌临床分离株的pbp2x基因和氨基酸序列的变异特点,探讨本地区的肺炎链球菌对青霉素及头孢噻肟的耐药机制.方法 2006年1月-2007年2月收集肺炎链球菌临床分离株34株,进行青霉素及头孢噻肟药敏试验,对青霉素不敏感的肺炎链球菌(PNSP)的青霉素结合蛋白pbp2x基因进行PCR扩增和测序,并进行BLAST分析.结果 有12株PNSP(青霉素及头孢噻肟MIC≥0.5 mg/L)发生了2个重要位点的氨基酸的替换:第一个保守基序STMK内Thr338→Ala及第三个保守基序KSG之前的Leu546→Val氨基酸替换.另外,菌株15发生了第二个保守基序SSN之前的His394→Leu氨基酸替换,而且本研究首次发现了紧邻第一个保守基序STMK后,Met342→Ile位点的氨基酸替换.有17个菌株的pbp2x基因的核苷酸及氨基酸序列出现了新的变异,已向GenBank提交,获得序列号:EU044831、EU089706-EU089709、EU106881-EU106884、EU124672.结论 本地区大多数PNSP的pbp2x核苷酸及氨基酸变异序列高度相似,提示肺炎链球菌对青霉素及头孢噻肟的耐药与pbp2x基因变异相关.     

病毒性肝炎后肝硬化住院患者血液感染大肠埃希菌超广谱β-内酰胺酶基因型调查CSCD

目的研究病毒性肝炎后肝硬化住院患者血液感染的大肠埃希菌产超广谱β内酰胺酶基因型。方法连续收集2011年1月至12月分离自病毒性肝炎后肝硬化住院患者血培养阳性的大肠埃希菌,VITEK-II鉴定细菌,K-β药敏试验检测细菌的药敏,多重PCR、PCR、测序及比对等对大肠埃希菌超广谱β内酰胺酶基因型进行分析。结果肝硬化主要原因为病毒感染,共收集79株大肠埃希菌,PCR扩增产物测序、比对：20株大肠埃希菌产TEM-1型β-内酰胺酶;仅有的-株产SHV-1型β-内酰胺酶,未检测出TEM型和SHV型超广谱β-内酰胺酶;40株产CTX-M型超广谱B-内酰胺酶,CTX-M-1群20株,CTX-M-9群26株,其中有6株菌同时产两群CTX-M型超广谱β-内酰胺酶,测序比对出CTX-M-3、CTX-M-15、CTX-M-14等八种基因型。结论病毒性肝硬化患者血液感染的大肠埃希菌超广谱B内酰胺酶最常见为CTX-M型,CTX-M-14是最常见的基因型。     

儿科产超广谱β-内酰胺酶志贺菌的基因型和耐药性研究

目的 探讨儿科临床分离志贺菌产超广谱β-内酰胺酶(ESBLs)的基因型及其耐药特点.方法 收集2004年1月至2008年12月北京儿童医院细菌性痢疾住院患儿粪便标本中分离出志贺菌共59株,按照美国临床和实验室标准协会推荐的表型确证试验检测ESBLs,用琼脂稀释法进行最低抑菌浓度(MIC)测定,对产ESBLs菌株进行PCR扩增明确其基因型,对扩增产物进行DNA序列分析确定基因亚型.结果 59株志贺菌中共检出产ESBLs者21株,占35.6%.21株产ESBLs志贺菌PCR均扩增到CTX-M型ESBLs,包括CTX-M-1型6株,CTX-M-9型15株,其中有4株同时伴有TEM型酶,6株伴有OXA型酶.DNA序列分析证实6株CTX-M-1型分别为CTX-M-3亚型(1株)、CTX-M-15亚型(2株)、CTX-M-57亚型(3株),15株CTX-M-9型均为CTX-M-14亚型,伴随存在的TEM型及OXA型耐药基因分别为TEM-1型广谱酶、OXA-1型广谱酶.药敏结果显示对产ESBLs菌株敏感性较好的抗生素有亚胺培南、美罗培南、哌拉西林/他唑巴坦、头孢哌酮/舒巴坦和头孢西丁,其耐药率均小于15%.产不同CTX-M基因亚型的菌株对头孢他啶的耐药性不同.结论 本地区儿科分离志贺菌产ESBLs阳性率高,均为CTX-M型,其中以CTX-M-14亚型为主,少部分为CTX-M-3、CTX-M-15和CTX-M-57亚型.大部分产ESBLs菌株呈多重耐药,碳青霉烯类抗生素应作为治疗产ESBLs志贺菌的首选.     

微生物耐药的分子检测现状

因临床工作和畜牧业过度使用抗菌素、抗生素 ,耐药微生物逐年增加。使得一些临床常见的感染性疾病成为难治性疾病 ,甚至一些耐药菌在医院内或局部区域内造成流行。2 0世纪 80年代以来 ,国外学者首先推出微生物耐药快速分子检测新技术 ,为患者能在第一时间明确诊断、合理治疗和疫情控制提供了有效手段。以下就微生物耐药分子检测的现状扼要简述。1.产超广谱 β-内酰胺酶菌产超广谱 β -内酰胺酶菌主要表现为对青霉素 ,头孢菌素Ｉ至ＩＩＩ类抗菌素的广泛耐药。常见为肺炎克雷伯氏菌和大肠埃希氏菌以及变形杆菌。由于头孢菌素已占医疗单位处方…     

Schizophrenia in a North Swedish geographical isolate, 1900–1977. Epidemiology, genetics and biochemistry

Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc² (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.     

Renal dysplasia and asplenia in two sibs

A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.     

The dominant diseases of modernized societies as omega-3 essential fatty acid deficiency syndrome: Substrate beriberi

About 1900, modern food selection and processing caused widespread $\text{epidemics}$ of the B vitamin deficiency diseases of beriberi and pellagra which, for genetic reasons, often expressed as different diseases ranging from bowel and heart disease to dermatoses and psychoses. But the B vitamins merely help convert essential fatty acids (EFA) into the prostaglandin (PG) tissue regulators and it now turns out that, through hydrogenation, milling and selection of w3-poor southern foods, we have also been systematically depleting, by as much as 90%, a newly discovered trace Nordic EFA (w3) of special importance to primates and sole precursor of the PG3(4) series, even as a concurrent fiber deficiency increases body demand for EFA. Since substrate EFA is processed by many B vitamin catalysts, an EFA deficiency will mimic a $\text{panh}$ypovitaminosis B, i.e., a mixture of $\text{substrate}$ beriberi and $\text{substrate}$ pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse $\text{endemic}$ disease. This would consitute a second stage of the Modern Malnutrition and explain why some workers now hold the dominant diseases of modermized societies to be new, nutritionally based, pellagraform yet lipid-related and to range, once again, from heart disease to psychosis. It is an assumption that our dominant diseases are unrelated to each other or are merely revealed by our diagnostic acumen and therapeutic success; and that hydrogenating millions of tons of food oils annually, to destroy the rancidity producing w3-EFA, is safe for $\text{primates}$. Extensive beriberiform disease is reported here in 32 typical cases taken from medical practice which responds strikingly to linseed oil supplements (60% w3-EFA) in confirmation of identical results in Capuchins.     

'Very sore nights and days': the child's experience of illness in early modern England, c.1580-1720

Sick children were ubiquitous in early modern England, and yet they have received very little attention from historians. Taking the elusive perspective of the child, this article explores the physical, emotional, and spiritual experience of illness in England between approximately 1580 and 1720. What was it like being ill and suffering pain? How did the young respond emotionally to the anticipation of death? It is argued that children’s experiences were characterised by profound ambivalence: illness could be terrifying and distressing, but also a source of emotional and spiritual fulfilment and joy. This interpretation challenges the common assumption amongst medical historians that the experiences of early modern patients were utterly miserable. It also sheds light on children’s emotional feelings for their parents, a subject often overlooked in the historiography of childhood. The primary sources used in this article include diaries, autobiographies, letters, the biographies of pious children, printed possession cases, doctors’ casebooks, and theological treatises concerning the afterlife.     

Guidelines for the Validation and Use of Immunoassays for Determination of Introduced Proteins in Biotechnology Enhanced Crops and Derived Food Ingredients

Recent advancements in agricultural biotechnology have created a need for analytical techniques to determine introduced proteins in crops enhanced through modern biotechnology techniques. These proteins are expressed in plant tissues and may be present in food ingredients. Immunoassays are ideally suited for protein detection and may be used as both quantitative and threshold methods. Microplate ELISA and lateral flow devices are two of the most commonly used immunoassay formats for agricultural biotechnology applications. This paper provides general background information and a discussion of criteria for the validation and application of immunochemical methods to the analysis of proteins introduced into plants and food ingredients using biotechnology methods. It is the result of a collaborative effort of members of the Analytical Environmental Immunochemical Consortium. This collaborative effort represents the combined expertise of several organizations to reach consensus on establishing guidelines for the validation and use of immunoassays. Further, the paper offers developers and users a consistent approach to adopting the technology as well as aid in producing accurate and meaningful results.     

Ultracryotomy: development and application (with examples from the yeast cytology) (author's transl)

The preparation steps usually necessary for obtaining ultrathin frozen sections of biological material (chemical prefixation, enclosing, cryoprotective treatment, freezing, sectioning, and post-staining the sections for transmission electron microscopy) are submitted to a critical analysis. The application of cryo-ultramicrotomy, in particularly for cytochemical purposes, is reviewed. Fundamental considerations of chemical prefixation and poststaining are supported by examples from yeast cytology. Furthermore, the efficiency of the cryo-ultramicrotomy (electron optical resolution of ultrastructural details) is demonstrated on yeast cells and protoplasts.     

HLA in North Colombia Chimila Amerindians

HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.     

The case for allele‐specific recognition by the TCR

There is a sharp difference in how one views TCR structure–function–behaviour dependent on whether its recognition of major histocompatibility complex‐encoded restriction elements (R) is germline selected or somatically generated. The generally accepted or Standard model is built on the assumption that recognition of R is by the V regions of the αβ TCR, which is not driven by allele specificity, whereas the competing model posits that recognition of R is allele‐specific. The establishing of allele‐specific recognition of R by the TCR would rule out the Standard model and clear the road to a consideration of a competing construct, the Tritope model. Here, the case for allele‐specific recognition (germline selected) is detailed making it obvious that the Standard model is untenable.     

The universal muscular chamber. A basic unit of the genitourinary, cardiovascular, gastro-intestinal, skeletal, cutaneous, respiratory and other systems, endocameral hypertension; and spasm, the key to their idiopathic diseases and arthropathies

Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.