DNA芯片检测肝组织及血清中乙型肝炎病毒DNA的临床研究

目的：研究DNA芯片检测乙型肝炎和肝硬化患者肝组织及血清中乙型肝炎病毒（HBV）DNA的应用价值。方法：用点样仪将聚合酶链反应（PCR）扩增的HBV DNA探针制成基因芯片。对15例慢性乙型肝炎病人的血清和肝活检组织，99例乙型肝炎后肝硬化肝组织，分别用基因芯片、原位分子杂交、免疫组织化学和雅培试剂检测HBV DNA、乙型肝炎核心抗原(HBcAg)、乙型肝炎表面抗原（HBsAg)和乙型肝炎e抗原（HBeAg)。结果：用基因芯片对15份HBsAg,HBeAg阳性的乙型肝炎患者血清进行检测，HBV DNA均阳性，阳性率符合率为100％；15份肝活检组织标本，免疫组化法检测HBcAg阳性15例，原位分子杂交法和基因芯片检测HBV DNA均阳性14例，阳性率93％。99份肝炎后肝硬化患者肝组织标本，HBcAg阳性67份，HBV DNA阳性53份，基因芯片检测HBV DNA阳性46份，阳性率分别为69％、88％。32例HBcAg、HBV DNA阳性的肝组织中，基因芯片检测HBV DNA均为阴性。结论：肝炎基因诊断芯片可检测肝组织及血清中HBV DNA，诊断准确率高，假阳性率低。     

乙型肝炎病毒基因型与临床关系的研究

[目的]研究乙型肝炎病毒（HBV）基因型与临床的关系。[方法]采用型特异性引物聚合酶链式反应PCR法对183例乙型肝炎患者的HBV进行基因分型，并对基因型与临床诊断、丙氨酸转移酶（ALT）、血清总胆红素（TBIL）、HBV DNA、T细胞亚群（CD4、CD8）水平及HBeAg、抗-HBe的关系进行了比较，应用SPSS 10.0 for windows软件整理分析数据。[结果]183例乙型肝炎患者中，急性乙型肝炎22（12.1％）例，慢性乙型肝炎81（43.0％）例，乙型肝炎后肝硬化57（31.8％）例，肝癌23（13.1％）例，其中B型41（19.2％）例，C型104（48.6％）例，B、C混合型38（17.8％）例。B、C、B＋C 3种基因型的患者ALT、TBIL以及CD4、CD8的水平进行比较，结果无显著性差异（P〉0.05）；而HBV DNA含量（对数值）比较有明显的差异（P〈0.05）；C基因型的HBeAg阳性率为75％，明显高于其他基因型（P〈0.05）。[结论]HBV基因型与乙型肝炎的临床进展有一定的相关性，急性乙型肝炎以B、C混合型为主，B型次之，慢性乙型肝炎、肝硬化、肝癌均以C型为主。基因型与ALT、TBIL、CD4、CD8的水平无关，但与HBV DNA水平有相关性，C基因型HBV DNA复制水平和HBeAg阳性率明显高于B基因型，预示C基因型可能与较严重的肝病发生有关。     

闽南地区乙肝病毒基因型与临床疾病的关系

目的 探讨闽南地区乙肝病毒基因型与临床疾病的关系.方法 采用实时荧光PCR法,对179例HBV感染者进行HBV基因分型,并检测血清HBV DNA、HBeAg及肝功能指标.结果 本地区HBV基因型以B型为主,其次为C型,少部分BC混合型.C型患者血清HBV DNA水平、HBeAg含量、HBeAg阳性率、肝功能指标均显著高于B型.结论 闽南地区HBV基因型以B型为主;C型HBV复制能力强,易引起较重肝损伤,与乙型肝炎病情加重有关.     

桂林地区乙肝患者HBV核苷类似物耐药位点分析

检测桂林地区乙型肝炎患者体内HBV逆转录酶基因的耐药突变情况,为制订合理的治疗方案提供依据.方法:提取乙肝患者血清中HBV DNA,巢式PCR扩增其逆转录酶基因后测序,Vector NTI Suite 8.0及chromaslite201分析测序结果,Mega4.0进行基因分型.结果:171例患者测序结果中18例发现有基因耐药突变,其中rtA181位点突变7例,占突变总数的38.9％.多药耐药基因突变12例,点突变总数的66.7％.B基因型患者耐药突变率为10.4％,C基因型患者耐药突变率为10.8％,两基因型分布比率的差异无统计学意义(P＞0.05).结论:桂林乙肝患者HBV逆转录酶基因耐药突变组合较为复杂,多药耐药基因突变比率较高.耐药突变的HBV基因型分布比率在所检测患者中无明显差异.     

构建乙型肝炎病毒基因参照品判断基因分型的探讨

目的利用乙型肝炎病毒（HBV）基因参照品判断HBV基因分型。方法自行制备HBV—S区基因参照品A、B、C、D型并测序及比较同源性，利用限制性内切酶MboI、EarI酶切参照品和样品扩增产物，从而确定样品基因型。利用该方法对克拉玛依地区272例HBV—DNA阳性血清基因分型。结果测序证实，HBV基因参照品与39例GenBank序列同源性均在96％以上。272例样品成功分型241例（88．6％）。检出B、C、D型，未检测到A型。抽检12例测序同源性大于96％。结论依据基因参照品核酸碱基片段大小判断分型，简明直观，适用于大样本的筛检及临床应用。     

简便HBV基因型S基因PCR-RFLP分型方法的建立

目的 建立简便HBV基因型S基因PCR-限制性片段长度多态性(RFLP)分型方法(以下简称“简便PCR-RFLP法”),并评价其临床应用价值.方法 临床诊断研究.查阅并比较GenBank中128株HBV(基因型A～D型)S基因片段(S区nt253-687),设计限制性内切酶Hinf Ⅰ、Ear Ⅰ、Apo Ⅰ鉴别A～D基因型分型方法,并评价简便PCR-RFLP法检测HBV DNA的最低检测下限、重复性;然后,用该方法检测50例慢性乙型重型肝炎患者的HBV基因型,同时用直接测序法验证简便PCR-RFLP法检测HBV基因型的一致性.结果 建立了简便PCR-RFLP法HBV基因分型的方案,通过限制性内切酶Hinf Ⅰ一次酶切就可分出我国流行的B型、C型、D型等毒株及B/C混合型.随机抽取3份标本,不同稀释浓度下,用简便PCR-RFLP法重复检测HBV基因型,分型结果均与原血清完全一致,且最低检测下限约为7 ～9 IU/ml.经Hinf Ⅰ酶一步酶切后,用简便PCR-RFLP法从50例患者标本中,检出B型23份、C型10份;经直接测序法验证,从PCR产物中检出B型23份、C型10份,2种方法检测B、C型基因结果完全一致(Kappa=1.00,P=0.001),简便PCR-RFLP法检出B/C混合型9份,优于PCR产物直接测序法(0份,x2=18.00,P=0.001).Hinf Ⅰ酶一步酶切后分出的ABD型8份,进一步酶切及应用PCR产物直接测序法检测均为B型变异株.结论 建立的简便PCR-RFLP法对HBV基因分型有较高的灵敏度和重复性,且在检出混合基因型方面具有优势,更为简便.     

浙江省义乌地区病毒性乙型肝炎病毒基因型分布

目的了解浙江省义乌地区病毒性乙型肝炎（乙肝）病毒（HBV）基因型分布特点,为乙肝的流行病学研究、临床诊治提供参考资料。方法用HBV基因分型（B、C基因型）荧光PCR法直接检测,或用HBV基因型特异引物经聚合酶链反应扩增乙肝患者血清HBV,将目的条带切下,经纯化后测序。测序得到的结果用CLUSTALX及Mega软件进行HBV分型分析。结果该地区基因分型结果,其中B型61.76% (63／102)、C型29.41％(30／102),B/C混合型8.82％ (9／102);男、女感染者中B型、C型基因型的构成比较,结果显示差异无统计学意义 (x2 =0.03,P＞0.05)。B/C混合基因型性别间差异有统计学意义 (x2=4.25,P＜0.05);对B基因型和C基因型感染者血清HBV-DNA定量检测结果比较,差异无统计学意义（Uc=1.1213,P＞0.05）,B基因型和C基因型感染者血清中HBeAg的阳性率分别是33.33%、20.00%,差异无统计学意义（x2 =1.75,P＞0.05）。结论浙江义乌HBV基因型为B型、C型和B/C混合型,以B型为主,且具有一定的地域特征。     

多对型特异性引物巢式PCR检测乙型肝炎病毒基因型分析

目的采用多对型特异性引物通过巢式PCR法检测HBeAg阳性患者血清中乙肝病毒(HBV)基因型的分布情况。方法根据从前S1基因到S基因中的保守序列设计出10条内外引物,并将其中8条内引物分成A、B两组分别扩增A、B、C和D、E、F型HBV,然后将第2轮PCR的两组产物分别用3%琼脂糖进行电泳,根据PCR产物片段大小直接判定HBV基因型。用该法检测15例慢性乙肝患者血清中HBV基因型,了解HBV基因型分布情况。结果 HBV基因分型结果为B型7例(46.7%)、C型4例(26.7%)、B+C型2例(13.3%),另有2例未能分型。结论人群HBV优势基因型以B型为主,C型次之。     

2型糖尿病患者中人类巨细胞病毒gH基因的酶切分型

目的 对2型糖尿病患者感染的人类巨细胞病毒(HCMV)进行gH基因检测和分型。方法 抽提101例2型糖尿病患者外周血白细胞DNA，以巢式PCR(nest PCR)法扩增HCMV gH基因，利用限制性核酸内切酶HhaI对HCMV gH基因进行限制性片段长度多态性分析(RFLP)。结果 16例2型糖尿病患者血标本为HCMV gH基因阳性，酶切分型结果均为gH1型。结论 杭州地区2型糖尿病患者HCMV流行株以gH1基因型为主，该基因型与标准株AD169有一定差异。     

西安市乙肝患者HBV基因型分布及临床相关性指标的研究

目的 研究了解西安市乙型肝炎病毒(HBV)的基因型分布及其与临床相关性指标丙氨酸氨基转移酶(ALT)和天门冬氨酸转氨酶(AST)关系和临床意义。方法 采用巢式PCR对389例西安市HBV感染患者的血清标本进行HBV DNA扩增,后用聚合酶链反应-限制性片段长度多态性法(PCR-RLFP)鉴定HBV基因型。并用酶联免疫吸附试验(ELISA)检测乙肝五项,速率法检测ALT和AST。结果 西安市389例HBV感染患者基因型分布为HBV B基因型58例占14.9%(58/389),HBV C基因型326例占83.8%(326/389),HBV B/C混合型5例占1.2%(5/389)。其中HBV B、C基因型与患者在性别和乙型肝炎e抗原(HBeAg)阳性率、HBV DNA载量水平及ALT和AST比较差异均无统计学意义(P0.05);B、C基因型患者在HBV携带者和慢性乙肝中分布比较差异无统计学意义(P0.05),但在肝硬化和肝癌中比较差异有统计学意义(P0.05)。HBV C基因型较B基因型更易发展成为肝硬化甚至肝癌。结论 西安市HBV感染者存在HBV B、HBV C、HBV B/C混合型3种基因型,以HBV C为主要基因型,B型次之,B/C混合型最少;肝硬化的发生与C型存在着一定关系,提示HBV C基因型更容易发展成为肝硬化。     

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.