甲基化寡核苷酸诱导灭活RASSFl基因在肝癌发病机制中的应用

目的:甲基化寡核苷酸诱导灭活RASSFl基因并探索其启动子甲基化在肝癌发病机制中的应用.方法:将与RASSF1基因启动子互补的甲基化寡核苷酸转染至肝癌SMMC-7721细胞系内,分别采用BSP和RT-PCR检测转染前后RASSFl基因启动子甲基化状态和mRNA表达状况.结果:正常SMMC-7721细胞系RASSF1基因启动子表现无甲基化状态并表达相应mRNA.转染互补甲基化寡核苷酸后,相应CG位点被诱导成完全甲基化状态.mRNA表达明显受到抑制.结论:互补甲基化寡核苷酸可诱导基因启动子甲基化状态.RASSF1基因启动子甲基化在原发性肝细胞癌发病机制中扮演重要角色.     

Runx3基因在肝细胞癌中甲基化状态分析及临床意义

目的通过检测肝细胞癌组织中Runx3基因启动子区域甲基化状态,探讨Runx3基因启动子区域异常甲基化在肝细胞癌发生、发展过程中的临床意义。方法采用DNA甲基化特异性聚合酶链反应（PCR）（MSP）技术对81例肝细胞癌患者肿瘤组织及其癌旁正常组织Runx3基因启动子区域甲基化状态进行检测。结果在肝细胞癌组织中,Runx3基因启动子区域甲基化率占43．2％（35／81）,而在相对应的癌旁正常组织中,Runx3基因启动子区域仅发现1例异常甲基化现象,其甲基化率占1．2％（1／81）（P〈0．01）;Runx3基因启动子区域甲基化状态与临床病理参数的关系差异均无统计学意义。结论Runx3基因启动子区域甲基化是导致Runx3基因失活的主要原因之一,与肝细胞癌发生早期密切相关,可作为肝细胞癌早期诊断的分子标记物及分子治疗的靶点。     

肝细胞癌多基因甲基化研究

目的 通过研究肝癌中多基因启动子区的甲基化,说明抑癌基因甲基化在肝癌发生的普遍性。 方法分别抽提冻存的肝癌、癌旁组织和正常肝组织的DNA,利用亚硫酸氢钠具有使未甲基化的胞嘧啶转为尿嘧啶而不改变甲基化胞嘧啶的特性分别设计甲基化引物和非甲基化引物,然后分别进行PCR 反应。我们分别检测了转移抑制基因p15, SYK, TIMP-3, E-cadherin,RASSAF1 和肿瘤相关基因p53, RB1, WT1, p14, p16启动子区在60个肝癌、癌旁和6个正常肝组织的甲基化情况。 结果60个肝癌标本中10个基因的甲基化情况各不相同:从p53的8%~90%的RASSAF1;肝癌组织比癌旁组织的甲基化平均水平高,二者在统计学上有显著差异（P<0.05）。 结论肝癌中有多个基因存在甲基化,且肝癌组织甲基化水平较癌旁组织高。      

甲基化寡核苷酸诱导灭活RASSF1基因在肝癌发病机制中的应用

目的:甲基化寡核苷酸诱导灭活RASSF1基因并探索其启动子甲基化在肝癌发病机制中的应用。方法:将与RASSF1基因启动子互补的甲基化寡核苷酸转染至肝癌SMMC-7721细胞系内,分别采用BSP和RT-PCR检测转染前后RASSF1基因启动子甲基化状态和mRNA表达状况。结果:正常SMMC-7721细胞系RASSF1基因启动子表现无甲基化状态并表达相应mRNA。转染互补甲基化寡核苷酸后,相应CG位点被诱导成完全甲基化状态,mRNA表达明显受到抑制。结论:互补甲基化寡核苷酸可诱导基因启动子甲基化状态,RASSF1基因启动子甲基化在原发性肝细胞癌发病机制中扮演重要角色。     

CDH11基因在肾细胞癌中的甲基化状态及其临床意义研究

目的检测CDH11基因启动子在肾癌中的甲基化情况,并分析甲基化与临床病理特征的关系。方法通过甲基化特异性PCR（MSP）检测CDH11基因在5株肾细胞癌细胞系、1株正常肾细胞系、46例肾细胞癌组织、23例癌旁肾组织,以及10例非肾实质肿瘤正常。肾组织中的甲基化状态。将甲基化情况与患者I临床资料联系,进行统计学分析。结果CDH11在5株肾癌细胞系中,有3株出现甲基化,甲基化率为60％;在人正常肾细胞系中未检测到甲基化。CDH11在肾癌组织中的甲基化率为45．7％（21／46）,显著高于癌旁肾组织（26．1％,6／23）及非肾实质肿瘤正常肾组织（0,（3／10）,差异具有统计学意义（P〈0．05）。肾细胞癌各分期间及各分级间,癌组织中CDH11基因甲基化检出率无统计学意义（P〉0．05）;左侧肾癌与右侧肾癌相比,癌组织CDH11基因甲基化率差异无统计学意义（P〉0．05）;男性肾癌患者与女性肾癌患者相比,肾癌组织中CDH1基因甲基化率差异无统计学意义（P〉0．05）。结论肾细胞癌组织中CDH11基因甲基化率显著高于癌旁正常肾组织及非肾实质肿瘤正常肾组织,且癌组织中CDH11基因甲基化率与临床病理资料如肿瘤分期及分级无显著相关性,提示CDH11基因甲基化是。肾细胞癌发生中的早期频发事件,可能是。肾细胞癌独特的基因甲基化谱成员之一,并在肾细胞癌的早期诊断上发挥作用。     

肝细胞癌中p14ARF基因启动子甲基化及蛋白表达的研究

目的 通过检测肝细胞癌中p14ARF基因启动子甲基化状态与蛋白表达的关系,探讨p14ARF基因启动子甲基化与肝细胞癌的关系.方法 通过甲基化特异性PCR(MSP)方法及免疫组化(IHC)法分别检测50例肝细胞癌组织和癌旁组织中p14ARF基因启动子甲基化状态和蛋白表达水平.结果 癌组织及癌旁组织中p14ARF基因启动子甲基化阳性率分别为28%(14150)和4%(2/50)(P<0.01).癌与癌旁组织p14ARF蛋白表达差异极显著(P<0.01).p14ARF基因启动子甲基化和蛋白表达与临床分期、门脉癌栓、术后复发、肝外转移、肿瘤大小、肿瘤个数及血清AFP值无关,而在肿瘤分化程度上p14ARF盯蛋白表达有显著差异,p14ARF基因启动子甲基化则无差异.p14ARF基因启动子甲基化与蛋白表达呈负相关.结论 启动子区甲基化是p14ARF基因失活的重要机制.p14ARF启动子区异常甲基化可能参与了肝癌的发生、发展,在肝癌的进展中起重要作用.     

应用BSP克隆测序检测肝细胞癌组织中Dynamin 3基因启动子区域甲基化水平

目的检测原发性肝细胞癌组织Dynamin 3(DNM3)基因启动子区域36个CpG的甲基化水平,分析其与肝细胞癌患者临床病理特征的关系,探讨DNM3基因甲基化在肝癌形成过程中的作用。方法提取30对患者肝细胞癌组织和癌旁组织DNA,经亚硫酸氢盐处理后,进行巢式PCR并联合克隆测序检测DNM3基因的甲基化水平。结果肝细胞癌患者的甲基化事件发生率为83.3%(25/30)。癌组织的DNM3基因启动子区域平均甲基化率为37.8%,癌旁组织为12.0%,两者比较差异有统计学意义(P0.05)。DNM3基因甲基化诊断肝癌的受试者工作特征曲线(ROC)曲线下面积(AUC)为0.831,对肝癌有较好的诊断价值。结论 DNM3作为肝细胞癌的一种抑癌基因,其启动子区域高度甲基化可能会促进肝细胞癌的形成。     

Runx3基因启动子区甲基化与胃癌的关系

目的通过检测胃癌Runx3基因启动子区域甲基化状态，来探讨Runx3基因启动子区域甲基化在胃癌发生和发展过程中的临床意义。方法采用DNA甲基化特异性PCR（MSP）技术对37例胃癌患者手术切除肿瘤组织及其癌旁组织Runx3基因启动子区域甲基化进行检测。结果Runx3基因在37例胃癌及其癌旁组织中甲基化率分别为40．5％（15／37）和8％（3／37）。结论Runx3基因启动子区甲基化可能是肿瘤特异性的（P＜0．05），可作为胃癌早期诊断的分子标记物。     

胃癌中多基因甲基化状态及其表达的研究

目的了解胃癌中p16、E-cadherin、Runx3、DAPK和CHFR基因启动子CpG岛的甲基化状态,并探讨基因异常甲基化与mRNA表达的关系及其意义。方法采用甲基化特异性PCR(MSP)技术检测12例胃癌患者癌组织及其相应的癌旁正常组织中5种基因的甲基化状态,以12例胃镜活检的正常胃组织作为对照,采用RT-PCR方法相应检测了5种基因的mRNA表达水平。并分析每种基因甲基化程度与表达水平之间的关系。结果胃癌组织中,p16、E-cadherin、Runx3、DAPK和CHFR基因的甲基化率分别为41.7%、8.3%、58.3%、33.3%和58.3%,相应的癌旁正常组织中,p16、E-cadherin、Runx3、DAPK和CHFR基因的甲基化率分别为8.3%、0.0%、8.3%、8.3%和16.7%,且75%的胃癌组织中至少有一种基因甲基化,显著高于癌旁正常组织25%(P<0.05),而健康对照组织中无基因甲基化,在甲基化的胃癌组织中均无p16、E-cadherin和DAPKmRNA表达,而癌旁正常组织、非甲基化的癌组织中均有其表达。结论胃癌中多基因启动子CpG岛高甲基化是一个频繁的事件,并且基因启动子高甲基化与其mRNA表达缺失有关,这可能参与了胃癌的发生发展。     

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.     

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.     

Clinical Pharmacokinetics of Morphine

Morphine, the most widely used mu-opioid analgesic for acute and chronic pain, is the standard against which new analgesics are measured. A thorough understanding of the pharmacokinetics of morphine is required in order to safely and effectively use this analgesic in a wide variety of patients with different levels of organ function. A MEDLINE search was conducted to identify literature published between 1966 and January 2002 relevant to the pharmacokinetics of morphine. These publications were reviewed and the literature summarized regarding unique and clinically important elements of morphine disposition relative to its parenteral administration (including intravenous, intramuscular, subcutaneous, epidural and intrathecal administration), absorption profile (immediate release, controlled release, and sublingual/buccal, and rectal administration), distribution, and its metabolism/ excretion. Special populations, including infants, elderly, and those with renal/liver failure, have a unique morphine pharmacokinetic profile that must be taken into account in order to maximize analgesic efficacy and reduce the risk of adverse events.     

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.     

Evidence-Based Pain Management and Palliative Care in Issue Four for 2009 of The Cochrane Library

ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly. Issue 4 for 2009 contains 4027 complete reviews, 1906 protocols for reviews in production, and 11447 one-page summaries of systematic reviews published in the general medical literature. In addition, there are citations of 600,000 randomized controlled trials, and 12,200 cited papers in the Cochrane methodology register. The health technology assessment database contains over 7500 citations. This edition of the Library contains 90 new reviews, of which 19 have potential relevance for practitioners in pain and palliative medicine.     

Wie zuverlässig sind Angaben zu nichtentzündlichen ausgedehnten Schmerzen?

ZusammenfassungFragestellung Es wurde geprüft, wie sich der Differenziertheitsgrad zweier Schmerzmessmethoden auf Angaben zur Ausgedehntheit klinischer Schmerzen auswirkt. Zugleich wurde der Referenzzeitraum variiert, über den die Patienten berichten sollten.Methode Erfasst wurde der Einfluss zu Lasten der Befragungsdifferenziertheit durch den Vergleich zweier Körperschema-Bildvorlagen. Drei Referenzzeiträume (Schmerz aktuell, letzte Woche, letztes halbes Jahr) wurden vorgegeben.Ergebnisse Patienten mit ausgedehnten Schmerzen gaben bei differenzierter Befragung um so mehr Schmerzen an, je weiter die Schmerzen zurück lagen und je größer der Berichtszeitraum war. Patienten mit gelenknahen Schmerzen gaben bei hoch differenzierter Befragung weniger ausgedehnte Schmerzen in der Vergangenheit an als bei globaler Einschätzung. Patienten mit Rückenschmerzen berichteten bei differenzierter Befragung zum aktuellen Schmerz über weniger ausgedehnte Schmerzen als bei globaler Befragung.Schlussfolgerung Die Angaben zur Schmerzausdehnung variieren vor allem bei Patienten mit ausgedehnten Schmerzen in Abhängigkeit von der Differenziertheit der Befragung. In diesen Fällen ist die Wahrscheinlichkeit erhöht, dass sich die Beschwerdesymptomatik zumindest teilweise erst in der Reaktion auf die situativen Befragungsbedingungen konstituiert und daher nicht auf andere Befragungsbedingungen generalisiert werden kann.