荧光定量PCR检测传染性单核细胞增多症不同血样本EBV的研究

[目的]寻找荧光定量PCR检测传染性单核细胞增多症（IM）EBV DNA拷贝量的最佳样本。[方法]采用荧光定量PCR分别检测了IM病人的外周血单个核细胞、全血基因组DNA及血浆的EBV DNA拷贝量,并比较各种方法的阳性率及拷贝量。[结果]三种不同来源的样品荧光定量PCR方法的阳性率分别为76％、69％、23％。全基因组及单个核细胞组阳性率比较经x^2检验,差异无显著性,且两组拷贝量比较差异亦无显著性。血浆组阳性率及拷贝量与前两种方法比较差异均有显著性。[结论]外周血单个核细胞是荧光定量PCR检测IM的EBV DNA量的最佳样本。     

肝硬化患者外周血单个核细胞中白蛋白mRNA的表达与病变进展的关系

目的：研究肝硬化患者外周血单个核细胞中白蛋白mRNA表达与病变进展的关系。方法：采用实时荧光定量PCR检测肝硬化患者血清中HBVDNA含量。采用逆转录巢式PCR分别检测代偿性肝硬化、失代偿性肝硬化和健康人外周血单个核细胞中白蛋白mRNA的表达。结果：37例肝硬化患者中21例为代偿期，16例为失代偿期。代偿期肝硬化患者血清HBVDNA阳性率为67％（14／21），HBVDNA含量为（4．3±2．1）×10^6 copies／mL：外周血单个核细胞白蛋白mRNA阳性率为14％（3／21）。失代偿期肝硬化患者血清HBVDNA阳性率为88％（14／16）．HBVDNA含量为（5．1±3．4）×10^7 copies／mL：外周血单个核细胞白蛋白mRNA阳性率为56％（9／16）。20例健康人外周血单个核细胞中的白蛋白mRNA阳性率为5％（1／20）。肝硬化代偿期和失代偿期患者外周血单个核细胞中的白蛋白mRNA阳性率差异有显著性（P〈0．1）。结论：HBVDNA与肝硬化患者肝脏病变进程密切相关，外周血单个核细胞中的白蛋白mRNA表达与肝脏损害相关。二者可作为肝硬化病情进展的评价指标。[著者文摘]     

巢式RT—PCR检测肺癌患者外周血微量转移癌细胞

[目的]建立检测肺癌患者外周血微转移癌细胞的敏感的分子检测方法，探讨其临床应用的意义。[方法]从肺癌患者外周血中分离有核细胞，用TRzol提取RNA，采用巢式反转录聚合酶链反应(RT—PCR)技术特异引物扩增CK19mRNA，凝胶电泳观察结果，以检测肺癌患者外周血CK19-mRNA的表达情况。分析CK19mRNA扩增结果与患者临床指标的关系。[结果]76例肺癌患者外周血标本中39例CK19mRNA阳性，阳性率为51．3％(39／76)，27例肺良性病变中仅1例阳性，阳性率为3．7％，15例健康对照者为阴性。[结论]巢式RT—PCR检测肺癌患者外周血微量转移癌细胞具有较高的敏感性和特异性，CK19-mRNA可作为一个标志物来检测肺癌患者外周血微转移。     

γδT细胞对系统性红斑狼疮患者外周血单个核细胞凋亡和CD69表达的影响

目的:研究γδT细胞对系统性红斑狼疮(SLE)患者外周血单个核细胞凋亡和CD69表达的影响,探索其在SLE发病中的调控作用。方法:采用单克隆抗体固相法,对8例正常人外周血γδT细胞进行体外扩增建系,并进行细胞纯度和表型鉴定;利用不同数量比的γδT细胞与15例SLE患者外周血单个核细胞进行体外共同培养3d,流式细胞仪检测单个核细胞凋亡和CD69表达的情况。结果:建立了正常人外周血γδT细胞系,其平均纯度为(80.3±9.2)%,细胞表型以γδ2[(86.1±8.0)%]和γγ9[(83.3±6.3)%]为主;SLE患者外周血单个核细胞的凋亡率在1∶100组[(2.48±3.86)%]和1∶50组[(1.17±1.38)%]均低于对照组[(3.77±3.96)%](P<0.05、P<0.01);γδT细胞对活动组和稳定组SLE患者外周血单个核细胞的凋亡均有明显的抑制作用(均P<0.05),但对两组之间的抑制强度无明显差异(P>0.05);CD69阳性率对照组、1∶100组和1∶50组之间差异无统计学意义(P<0.05)。结论:γδT细胞体外能够抑制SLE患者外周血单个核细胞的凋亡,且这种抑制作用与狼疮的活动性无关,与外周血细胞的活化无关。     

T-SPOT.TB在肺部疾病结核感染筛查中的应用

目的 评价结核分枝杆菌(后简称结核杆菌)感染T细胞斑点试验(T-SPOT.TB)诊断肺部疾病感染结核的价值。方法回顾性分析405例肺部疾病患者资料,所有患者均行外周血T-SPOT.TB,血清结核抗体(TB-Ab)、痰液涂片抗酸菌检测,结核DNA检测,并对其检测结果进行分析,比较各指标的阳性率。结果T-SPOT.TB阳性率为39.26%,远高于TB-Ab(10.86%)、痰液涂片(1.23%)、TB-DNA(3.95%)的阳性率。结论 T-SPOT.TB在结核病中具有较好的辅助诊断价值,其敏感性显著高于其他检测方法,可有效检出结核潜伏感染,帮助临床鉴别诊治疾病,具有较高推广应用价值。     

PCR-微板杂交-ELISA检测结核杆菌DNA

[目的]探讨PCR-微板杂交-ELISA方法检测结核杆菌DNA的临床价值。[方法]选择PCR扩增区域内的保守序列设计捕获探针和显色探针,分别用于DNA杂交和ELISA分析,建立PCR-微板杂交-ELISA方法并运用该方法对328份结核病人痰标本进行了检测。[结果]328份结核病人痰标本中有196例结核杆菌DNA阳性,阳性率为59．7％。检出率明显高于痰培养和痰涂片抗酸染色。[结论]PCR-DNA微板杂交-ELISA方法综合运用了PCR、DNA杂交和-ELISA技术,大大增加了DNA检测的特异性和灵敏度,在临床上具有良好的应用前景。     

一种简单快速扩增和获取外周血γδT细胞的方法

本研究目的是建立一种简单快速的扩增人外周血γδT细胞的方法，以便用于γδT细胞的生物学特性和临床过继性免疫治疗的研究。取人外周血5-10ml，分离获取单个核细胞，用结核杆菌低分子多肽抗原(MTb-Ag)刺激细胞增殖，通过荧光单克隆抗体TCR γδ-PE染色及流式细胞仪检测γδT细胞所占比例，用MTT试验检测细胞毒活性。结果表明：MTb-Ag能特异地刺激外周血单个核细胞中的γδT细胞增殖，γδT细胞可达69.2％，对K562细胞具有较高的杀伤活性。结论：本方法是一种简单、快速、特异的体外扩增和获取人外周血γδT细胞的方法。     

单细胞人白细胞抗原多基因位点检测的配型预选性研究及意义

目的　探索单细胞扩增前引物延伸法 (PEP)全基因组扩增 ,后续多重巢式聚合酶链反应 (MN PCR)同步扩增人类白细胞抗原 (HLA) A、B和DR基因位点检测技术 ,进行配型预选研究的价值。方法　采用PEP MN PCR方法测定单个淋巴细胞和单个胚胎细胞的HLA A、B和DR基因型。结果　PEP MN PCR对单个淋巴细胞的扩增率、扩增准确率、假阴性率和假阳性率分别为 91 7%、95 7%、3 0 %和 1 7% ,而单个胚胎细胞则分别为 97 7%、97 5 %、2 3%和 2 6 % ,两种细胞检测结果比较差异均无显著性意义 (P >0 0 5 )。结论　单细胞PEP MN PCR检测HLA多基因位点有较高的扩增效率和扩增准确率 ,具有应用于植入前胚胎遗传学分析 ,筛选HLA匹配胚胎 ,提供脐血造血干细胞移植治疗同胞疾病的应用价值。     

荧光定量PCR技术在结核杆菌检测中的应用

目的：建立结核病荧光定量PCR快速检测方法。方法：建立检测结核杆菌的荧光定量PCR方法并进行特异性、敏感性、重复性实验，对临床样品进行检测分析。结果：该方法的敏感性达到了1Pg，且特异性高．重复性好。应用荧光定量PCR法及抗酸染色、结核杆菌培养法，对332例活动性肺结核患者晨痰、215例活动性肺结核患者外周血、187例结核性胸膜炎患者胸水进行检测．荧光定量PCR技术阳性率分别为53．0％（176／332）、34．4％（74／215）、36．9％（69／187），经统计学比较分析，3种方法检测率差异有统计学意义（P〈0．01）。结论：荧光定量PCR技术具有简便、快速、防污染、敏感性与特异性高等优点，是结核病诊断的有效方法之一。[著者文摘]     

380例全血细胞减少患者血片与骨髓象检查结果分析

[目的]了解全血细胞减少患者的血片幼稚细胞检出与骨髓象分析情况。[方法]对380例全血细胞减少患者进行外周血涂片及骨髓象分析。[结果]380例全血细胞减少患者中，血片检出幼稚细胞81例，其中急性白血病（AL）71例，骨髓异常增生综合征（MDS）患者8例，巨幼红细胞性贫血（MA）3例。未检出幼稚细胞患者为299例。骨髓检查主要以再障（AA）为主，其次为巨幼红细胞性贫血（MA）、慢性肝病、恶性肿瘤、感染及其他疾病。其中急性白血病5例，MDS患者54例。[结论]全血细胞减少患者外周血片幼稚细胞检出率主要与血液系统疾病有关，患者宜早行骨髓象分析确诊。骨髓象分析是明确全血细胞减少病因的重要方法。     

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.