白假丝酵母临床分离株基因分型与药敏分析

目的分析上海交通大学医学院附属瑞金医院白假丝酵母的基因分型情况以及不同基因型白假丝酵母的药敏结果。方法采用聚合酶链反应（PCR）扩增白假丝酵母25S核糖体DNA基因的内含子区，可转座Ⅰ型内含子插入片段的数目及大小的不同使得扩增产物不同，根据产物片段大小和数目分型。采用微量稀释法分析白假丝酵母抗真菌药物敏感性。结果171株白假丝酵母分为3型：A型93株，B型47株，C型31株。B型、C型白假丝酵母对唑类抗真菌药物的耐药率显著高于A型菌株，对氟胞嘧啶的耐药率低于A型菌株。结论白假丝酵母的PCR分型方法简便、快速、重复性好、特异性强；不同型别的白假丝酵母耐药谱有差异，对治疗中抗真菌药物的选择有一定的指导意义。     

获得性免疫缺陷综合征患者白假丝酵母分离株基因型及耐药性分析

目的研究获得性免疫缺陷综合征(AIDS)患者白假丝酵母分离株的基因型及耐药性。方法对分离自上海市公共卫生中心AIDS住院患者的40株白假丝酵母,应用微卫星核心序列引物M13进行聚合酶链反应(PCR)指纹分型。用微量稀释法分析白假丝酵母抗真菌药物敏感性。结果PCR指纹分型将所有白假丝酵母分离株分为A、B、C、D 4种基因型,其中A型14株(35.0%),B型16株(40.0%),C型9株(22.5%),D型1株(2.5%)。白假丝酵母对抗真菌药物的耐药率为:两性霉素B 2.5%,氟康唑22.5%,伊曲康唑15.0%,氟胞嘧啶20.0%,;不同基因型白假丝酵母菌株间耐药性差异无统计学意义。结论上海市AIDS住院患者分离的白假丝酵母主要由3种克隆组成,但无明显优势流行株。     

获得性免疫缺陷综合征患者白假丝酵母分离株基因型及耐药性分析

目的研究获得性免疫缺陷综合征（AIDS）患者白假丝酵母分离株的基因型及耐药性。方法对分离自上海市公共卫生中心AIDS住院患者的40株白假丝酵母,应用微卫星核心序列引物M13进行聚合酶链反应（PCR）指纹分型。用微量稀释法分析白假丝酵母抗真菌药物敏感性。结果PCR指纹分型将所有白假丝酵母分离株分为A、B、C、D 4种基因型,其中A型14株（35.0%）,B型16株（40.0%）,C型9株（22.5%）,D型1株（2.5%）。白假丝酵母对抗真菌药物的耐药率为：两性霉素B 2.5%,氟康唑22.5%,伊曲康唑15.0%,氟胞嘧啶20.0%,;不同基因型白假丝酵母菌株间耐药性差异无统计学意义。结论上海市AIDS住院患者分离的白假丝酵母主要由3种克隆组成,但无明显优势流行株。     

耐甲氧西林金黄色葡萄球菌医院感染爆发流行的实验室调查

目的采用耐甲氧西林金黄色葡萄球菌(MRSA)mecA基因相关高变区(HVR)长度多态性分型了解MRSA的医院感染爆发流行情况。方法选取临床分离的38株MRSA菌株,采用聚合酶链反应(PCR)方法扩增MRSA mecA基因HVR,据扩增片段大小差异进行基因分型。结果根据PCR产物片段大小,38株MRSA被分为A、B、C、D 4个基因型,其中C型23株(60.53%)、A型7株(18.42%)、B型3株(7.89%)、D型2株(5.26%)、未分型3株(7.89%)。C型在临床各科室均有分布,但主要集中于老干部科。MRSA呈严重且多重耐药,故首选抗菌药物为万古霉素、替加环素、奎奴普汀/达福普汀和替考拉宁。结论医院内存在着MRSA菌株的克隆传播,应加强检测,控制医院感染爆发流行。     

白色假丝酵母菌ERG11基因突变与唑类抗真菌药物耐药的关系

目的探讨白色假丝酵母菌ERG11基因突变与唑类抗真菌药物耐药的关系。方法纸片扩散法初步筛选临床分离的耐唑类抗真菌药物的白色假丝酵母菌,测定初筛耐药株对氟康唑和伊曲康唑的最低抑菌浓度,随机选择10株白色假丝酵母耐药株,提取基因组DNA。利用DNASTAR软件辅助设计3对PCR引物,以提取的目的 DNA为模板,分别从前(P1)、中(P2)、后(P3)3段扩增ERG11基因全序列。扩增后的PCR产物经纯化后测序,将测序结果与GenBank中已知标准序列(X13296)进行比较分析。结果电泳结果显示,获得的3段PCR产物大小与预期结果一致。测序结果显示成功获得10株白色假丝酵母菌ERG11全基因序列。与GenBank中标准株序列(X13296)比较,耐药株均存在同义突变和错义突变,共27个碱基突变位点。15个位点是错义突变,其中F126LI、166N、H183Q、V437I、S453F和N490K为新发现的突变位点。结论耐唑类抗真菌药物的白色假丝酵母菌ERG11基因有多个发生错义突变的位点,且为多点突变,可能与其耐药有关。     

中国汉族人群中白色念珠菌的25S rDNA基因分型及耐药性分析

目的调查白色念珠菌25S核蛋白体脱氧核糖核酸(rDNA)基因型分布,并分析白色念珠菌各基因型与耐药性之间的相关性。方法以经API 20C AUX真菌鉴定板和科玛嘉念珠菌显色培养基鉴定到种的76株白色念珠菌为研究对象,转沙保罗氏培养基纯培养24 h,收集菌液,裂解法提取质粒DNA,用特异性引物扩增白色念珠菌25S rDNA基因,根据聚合酶链反应(PCR)产物电泳图谱进行基因分型;采用微量肉汤稀释法测定各基因型白色念珠菌对4种常用抗真菌药物5-氟胞嘧啶(5-FC)、两性霉素B(AMB)、氟康唑(FCZ)、伊曲康唑(ICZ)的最低抑菌浓度(MIC)。结果 76例白色念珠菌分为A、B、C 3个基因型(A型48例,B型8例,C型20例),按照有无长度为379 bp的Ⅰ型内含子的插入,将76例白色念珠菌分为有内含子组和无内含子组,分别进行4种抗真菌药物耐药率的比较,结果发现2组间对FCZ耐药率差异有统计学意义(P0.01),而对5-FC、AMB和ICZ耐药率差异无统计学意义(P值均大于0.05)。结论白色念珠菌25S rDNA基因内含子(379 bp)的缺失或者插入可能与其对FCZ的耐药性有关。     

不可分群脑膜炎奈瑟菌PCR基因分型研究

目的对自健康人群鼻咽拭子中分离的74株表型特征为不可分群的脑膜炎奈瑟菌,利用PCR基因分型技术进行基因特征分析。方法用PCR方法扩增脑膜炎奈瑟菌种属特异性基因crgA和A、B、C、W135、Y、Z、X以及29E不同群的特异性基因片段,确定不同菌株的基因型。结果以脑膜炎奈瑟菌crgA基因为靶基因进行PCR扩增,74株不可分群脑膜炎奈瑟菌中,66株脑膜炎奈瑟菌crgA基因扩增阳性,检出率89.19%。66株菌株进行不同血清群的PCR分群鉴定,40株菌株（60.61%）可检测为不同的血清群基因型,B群27株,29E群7株,X、Y群各2株,C、W135各1株。结论不可分群脑膜炎奈瑟菌进行PCR基因分型研究对脑膜炎奈瑟菌的研究具有重要的流行病学意义。     

变性高效液相色谱对CTX-M型超广谱β内酰胺酶基因分型研究

目的 探讨湖南省CTX-M型超广谱B内酰胺酶(ESBLs)基因型分布情况和变性高效液相色谱(DHPLC)方法检测CTX-M型ESBLs基因型的准确性.方法 用多重PCR扩增标准菌株和ESBLs表型阳性的临床菌株blaCTX-M基因,扩增产物经DHPLC分析得到标准菌株和临床菌株色谱峰图,通过比对标准色谱峰图对临床菌株进行基因分型,同时采用单纯随机抽样法选择25株多重PCR扩增阳性菌株进行特异PCR扩增,其产物冉进行基因测序来评估DHPLC法的准确性.结果 142株产ESBLs的肠杆菌科细菌经多重PCR扩增证实109株携带blaCTX-M基因,检出率为76.8%(109/142).109株扩增阳性的菌株经DHPLC分析后检出4种不同的blaCTX-M基因型:33株携带CTX-M-3、19株携带CTX-M-15、5株携带CTX-M-9、52株携带CTX-M-14.25株基因测序结果与DHPLC基因分型结果作比较显示:24株DHPLC的基因分型结果与基因测序结果完全吻合,但有1株DHPLC基因分型为CTX-M-15,而测序为CTX-M-82.结论 DHPLC可对耐药进行基因快速基因分型,具有准确、快速和经济等优点.     

比较重复序列PCR与多位点序列分型技术用于光滑假丝酵母菌的基因分型检测

目的 评价REP-PCR基因分型技术在临床光滑假丝酵母菌株分型中的应用价值。方法收集2009-2010年上海瑞金医院、上海仁济医院、上海华山医院、安徽医科大学附属医院、深圳人民医院38株光滑假丝酵母菌。采用MLST法扩增光滑假丝酵母菌的6个管家基因内片段,扩增片段测序后与数据库数据比对得出等位基因谱,从而得到相应的序列型（sequence type,ST）。采用REP-PCR法设计Ca21、Ca22、Com21引物扩增38株光滑假丝酵母菌重复序列,通过电泳比较分析,获得REP-PCR型。比较两种分型方法的结果和效率。结果 以Ca22-Com21为引物的REP-PCR分型效果最好。REP-PCR和MLST分型结果相同,38株光滑假丝酵母菌共产生5种REP-PCR型,分别为A、B、C、D、E型,对应于MLST的ST7、ST3、ST19、ST45和新型。REP-PCR比MLST耗时短。结论REP-PCR方法简单快速,且分辨率与MLST技术相同,可作为实验室大量菌株分型的首选方法。     

婴幼儿感染猪霍乱沙门菌耐药基因特点及同源性分析

目的研究致婴幼儿败血症猪霍乱沙门菌的耐药基因特点及同源性。方法16株猪霍乱沙门菌．采用K—B纸片扩散法检测抗菌药物的耐药性,同时用PCR法及DNA测序法对其进行β-内酰胺酶类耐药基因检测;PFGE法测定同源性。结果对氨苄西林耐药的13株猪霍乱沙门菌经PCR扩增后并测序为bl岫。16株猪霍乱沙门菌经PFGE分型,可分为5个PFGE型,其中A型是最主要的克隆（12／16）。13株blTEM-1．基因阳性菌株分别为A型11株,B型1株,D型1株。结论产TEM-1割β-内酰胺酶是本地区猪霍乱沙门菌对氨苄西林耐药的主要机制,PFGE分型方法对猪霍乱沙门菌分型能力较好。克隆性传播为猪霍乱沙门菌的主要传播途径．同一PFGE型菌株的耐药谱非常接近。     

Determination of creatine kinase isoenzyme MB activity in serum using immunological inhibition of creatine kinase M subunit activity. Activity kinetics and diagnostic significance in myocardial infarction.

This is a new method for the determination of creatine kinase isoenzyme MB activity in serum. The method uses direct activity measurement of creatine kinase B subunit activity after blocking of CK-M subunit activity by inhibiting antibodies. The test takes no longer than 15 min. The method yields an intra-serial C.V. of 2.0-12.9%, and a C.V. from day to day of 5.5%. The detection limit is 3.4 U/l creatine kinase MB. In the 95 cases with proven myocardial infarction several types of creatine kinase MB activity kinetics could be determined. The percentage of creatine kinase MB of peak CK-total is 6-25%, with a mean of 11.1%. The amount of creatine kinase MB with respect to total CK activity after reinfarction is higher than the amount after initial infarction.     

Dissociable correlates of two classes of retrieval processing in prefrontal cortex

Although substantial evidence suggests that the prefrontal cortex (PFC) implements processes that are critical for accurate episodic memory judgments, the specific roles of different PFC subregions remain unclear. Here, we used event-related functional magnetic resonance imaging to distinguish between prefrontal activity related to operations that (1) influence processing of retrieval cues based on current task demands, or (2) are involved in monitoring the outputs of retrieval. Fourteen participants studied auditory words spoken by a male or female speaker and completed memory tests in which the stimuli were unstudied foil words and studied words spoken by either the same speaker at study, or the alternate speaker. On "general" test trials, participants were to determine whether each word was studied, regardless of the voice of the speaker, whereas on "specific" test trials, participants were to additionally distinguish between studied words that were spoken in the same voice or a different voice at study. Thus, on specific test trials, participants were explicitly required to attend to voice information in order to evaluate each test item. Anterior (right BA 10), dorsolateral prefrontal (right BA 46), and inferior frontal (bilateral BA 47/12) regions were more active during specific than during general trials. Activation in anterior and dorsolateral PFC was enhanced during specific test trials even in response to unstudied items, suggesting that activation in these regions was related to the differential processing of retrieval cues in the two tasks. In contrast, differences between specific and general test trials in inferior frontal regions (bilateral BA 47/12) were seen only for studied items, suggesting a role for these regions in post-retrieval monitoring processes. Results from this study are consistent with the idea that different PFC subregions implement distinct, but complementary processes that collectively support accurate episodic memory judgments.     

俯卧位通气对高海拔地区肺复张治疗无效急性呼吸窘迫综合征患者氧合的影响

目的 探讨俯卧位通气对高海拔地区肺复张术(RM)治疗无效急性呼吸窘迫综合征(ARDS)患者的治疗作用.方法 从海拔2260m的地区医院筛选RM治疗无效的41例ARDS患者[平均氧合指数( PaO2/FiO2)较RM前升高＜20％视为RM无效],依不同病因分为肺内源性ARDS组(ARDSp组)和肺外源性ARDS组(ARDSexp组),每组再按信封法随机分为俯卧位组和仰卧位组,即ARDSp俯卧位组(11例)、ARDSp仰卧位组(9例)、ARDSexp俯卧位组(10例)、ARDSexp仰卧位组(11例).在通气前及通气1、2、3、4h监测动脉血氧分压( PaO2)、PaO2/FiO2、静态顺应性(Cst)、气道阻力(Raw)的变化.结果 通气lh时,ARDSexp俯卧位组PaO2/FiO2( mm Hg,l mm Hg=0.133 kPa)即较通气前显著升高(157.4±40.6比129.3±48.7,P＜0.05),并随通气时间延长呈持续增高趋势,4h达峰值(219.1 ±41.1);且ARDSexp俯卧位组通气3h内PaO2/FiO2较其他3组显著增高,另3组间则差异无统计学意义.ARDSp俯卧位组、ARDSexp俯卧位组通气4h时PaO2/FiO2均较相应仰卧位组显著增高(208.8±39.7比127.4±47.1,219.1±41.1比124.9±50.8,均P＜0.05).4组通气前后Cst无显著改变,各组间差异也无统计学意义.ARDSp俯卧位组通气4h时Raw(cmH2O·L-1·s-1)较通气前显著降低(6.8±1.7比10.7±1.8,P＜0.05),且明显低于其他3组;其他3组各时间点Raw组内及组间比较差异均无统计学意义.结论 俯卧位通气作为ARDS机械通气重要策略之一,可以改善RM无效高原ARDS患者的氧合,为抢救患者赢得宝贵的时间.     

Digital imaging among medical facilities

The Department of Veterans Affairs (VA) in the USA operates a network of 172 medical centres which all utilize a hospital information system (HIS) which has been developed and is currently maintained by the VA. During the past several years, an image management and communication module has been developed, installed and clinically utilized at the Washington DC and Maryland VA Medical Centres. This image management and communication system, referred to as the decentralized hospital computer program (DHCP) imaging system, is fully integrated with a commercial picture archiving and communication system (PACS). The system is utilized to capture, archive, and display all images generated within the hospital including radiology, nuclear medicine, pathology, endoscopy, bronchoscopy, and dermatology, intraoperative photographs, ECG data, and a limited number of paper documents. The ultimate goal of the project is to have all patient text and image data available at any clinical workstation to any authorized user anywhere within the network of medical centres. Clinical requirements for an imaging workstation include ease of use, rapid and reliable access to the complete set of patient information, and images which are of acceptable quality to meet the requirements of the user and the subspecialty. Patient confidentiality and data security must be safeguarded at all times. Integration of the images with the remainder of the patient's database was found to be critical to the success of the project. The experience at the Washington and Maryland facilities suggests that an imaging system that is successfully integrated with a hospital information system can provide substantial clinical and economic benefits both within and among medical centres. Clinical acceptance and utilization of the system has been excellent, particularly in diagnostic radiology where DHCP Imaging has been interfaced to a commercial PAC system. Based upon this initial experience, the VA has begun to deploy the system throughout its large network of medical centres.     

Myocardial elastography at both high temporal and spatial resolution for the detection of infarcts

Myocardial elastography is a novel method for noninvasively assessing regional myocardial function, with the advantages of high spatial and temporal resolution and high signal-to-noise ratio (SNR). In this paper, in-vivo experiments were performed in anesthetized normal and infarcted mice (one day after left anterior descending coronary artery [LAD] ligation) using a high-resolution (30 MHz) ultrasound system (Vevo 770, VisualSonics Inc., Toronto, ON, Canada). Radiofrequency (RF) signals of the left ventricle (LV) in longitudinal (long-axis) view and the associated electrocardiogram (ECG) were simultaneously acquired. Using a retrospective ECG gating technique, 2-D full field-of-view RF frames were acquired at an extremely high frame rate (8 kHz) that resulted in high-quality incremental displacement and strain estimation of the myocardium. The incremental results were further accumulated to obtain the cumulative displacements and strains. Two-dimensional and M-mode displacement images and strain images (elastograms), as well as displacement and strain profiles as a function of time, were compared between normal and infarcted mice. Incremental results clearly depicted cardiac events including LV contraction, LV relaxation and isovolumetric phases in both normal and infarcted mice, and also evidently indicated reduced motion and deformation in the infarcted myocardium. The elastograms indicated that the infarcted regions underwent thinning during systole rather than thickening, as in the normal case. The cumulative elastograms were found to have higher elastographic SNR (SNR(e)) than the incremental elastograms (e.g., 10.6 vs. 4.7 in a normal myocardium, and 6.0 vs. 2.4 in an infarcted myocardium). Finally, preliminary statistical results from nine normal (m = 9) and seven infarcted (n = 7) mice indicated the capability of the cumulative strain in differentiating infracted from normal myocardia. In conclusion, myocardial elastography could provide regional strain information at simultaneously high temporal (>/=0.125 ms) and spatial ( approximately 55 microm) resolution as well as high precision ( approximately 0.05 microm displacement). This technique was thus capable of accurately characterizing normal myocardial function throughout an entire cardiac cycle, at the same high resolution, and detecting and localizing myocardial infarction in vivo.     

手转胎头术失败的原因与分娩结局临床分析

目的 探讨手转胎头术失败的原因与分娩结局.方法 选择2008年1月至2010年12月于我院住院分娩的持续性枕横位、枕后位产妇198例,根据行手转胎头术后结果分为成功组126例、失败组72例.比较两组分娩结局,对比分析失败原因.结果 失败组胎儿体质量≥3500 g的发生率[76.4％(55/72)]明显高于成功组[31.7％(40/126)],差异有统计学意义(x2=30.177,P=0.001)、失败组宫缩乏力发生率[58.3％(42/72)]高于成功组[38.1％ (48/126)],差异有统计学意义(x2=7.569,P=0.006)、失败组骨盆临界或轻度狭窄发生率[38.9％ (28/72)]高于成功组[23.8％(30/126)],差异有统计学意义(x2 =5.030,P=0.002)、失败组手转胎头时机不当(宫口开大＜6 cm、胎头位于坐骨棘上及宫口开大8～10 cm、胎头位于坐骨棘下≥2 cm)发生率[61.1％(44/72)]高于成功组[38.9％(49/126)],差异有统计学意义(x2=9.084,P=0.003).失败组母儿并发症(产后出血、产褥病率、胎儿窘迫、新生儿窒息)发生率高于成功组(x2 =9.586,P=0.002、x2=9.334,P=0.002、x2=5.910,P=0.015、x2=5.240,P=0.022)、失败组剖宫产发生率[72.2％(52/72)]明显高于成功组[34.1 ％(43/126),x2=26.641,P=0.001)].结论 手转胎头术能使难产变顺产,降低剖宫产率,减少母儿并发症,但须积极预防、处理导致手转胎头术失败的原因,对矫正失败后继续矫正及试产应慎重.