骨髓增生异常综合征患者SHP-1基因启动子甲基化状态及相关机制的探讨

目的　探讨SHP-1基因异常甲基化在骨髓增生异常综合征(MDS)发生、发展中的作用和相关机制.方法　将63例MDS患者根据IPSS积分系统分为较低危和较高危组.采用甲基化特异性PCR(MS-PCR)对MDS细胞系SKK-1细胞和MDS患者骨髓细胞进行SHP-1基因启动子甲基化状态的检测;用Western blot方法检测正常对照者和患者骨髓单个核细胞和SKK-1细胞磷酸化信号传导和转录激活因子3( p-STAT3)蛋白的表达.结果　SKK-1细胞存在SHP-1基因启动子部分甲基化,正常对照者骨髓细胞均为非甲基化,较高危组MDS患者SHP-1启动子甲基化率显著高于较低危组(63.3％对24.2％)(P＜0.05);按WHO分型中染色体核型和骨髓原始细胞比例分组,SHP-1启动子甲基化率在RAEB-Ⅱ、核型差组和骨髓原始细胞占0.11 ～0.19组显著增高(P＜ 0.05);SKK-1细胞中存在p-STAT3蛋白的表达,较高危组MDS患者p-STAT3蛋白的表达显著高于较低危组(66.7％对18.2％);相关分析显示SHP-1启动子甲基化的出现与p-STAT3蛋白的表达有相关性.结论　SHP-1基因启动子的异常甲基化在MDS的发病和疾病进展中发挥作用,其机制可能涉及STAT3信号通路的激活,检测MDS患者SHP-1启动子甲基化状态有助于对疾病预后的判断.     

骨髓增生异常综合征患者血浆DNA中FHIT基因启动子区甲基化状态及地西他滨的去甲基化作用

本研究旨在检测骨髓增生异常综合征(MDS)患者血浆DNA中FHIT基因启动子区域甲基化状况及地西他滨对其甲基化的影响。采用甲基化特异性聚合酶链反应法检测1例初治的MDS患者、3例MDS转化而来的AML患者在地西他滨序贯半量CAG方案化疗前后血浆DNA中FHIT基因启动子区域CPG岛甲基化情况,并分析其临床疗效。结果表明,3例患者治疗前有FHIT基因甲基化。治疗1个疗程后其中2例患者FHIT基因甲基化得到逆转,4例患者中有2例获得临床缓解,2例无效。结论:MDS的发生可能与FHIT基因甲基化相关,地西他滨对MDS患者血浆DNA中FHIT基因高甲基化具有明显的去甲基化作用。血浆DNA的FHIT基因甲基化检测可能成为MDS辅助诊断和预后判断的分子标记。     

髓系白血病细胞中细胞因子信号转导抑制因子1基因启动子去甲基化对细胞增殖、凋亡的影响

目的 探讨髓系白血病(ML)细胞中细胞因子信号转导抑制因子1(SOCS-1)基因启动子去甲基化对细胞增殖、凋亡的影响。方法 收集78例ML患者的骨髓标本(实验组)以及人ML细胞系U937、HL-60、K562为研究对象,另外收集50例健康捐献者的骨髓标本作为对照组。甲基化特异性聚合酶链反应(MS-PCR)检测ML患者骨髓标本中SOCS-1基因启动子甲基化状态,实时荧光定量聚合酶链反应(qRT-PCR)检测ML患者骨髓标本以及人ML细胞系中SOCS-1 mRNA表达;Western blot检测ML患者骨髓标本以及人ML细胞系中SOCS-1蛋白表达。用0、0.8、1.6、3.2μmol/L地西他滨(DCA)分别处理U937细胞,并分别命名为空白组、DCA-L组(DCA低剂量)、DCA-M组(DCA中剂量)、DCA-H组(DCA高剂量)。MS-PCR检测各组细胞中SOCS-1基因启动子甲基化状态;qRT-PCR检测各组细胞中SOCS-1 mRNA表达;Western blot检测各组细胞中SOCS-1蛋白表达;CCK-8法检测各组细胞增殖情况;流式细胞术检测各组细胞凋亡情况。结果 SOCS...     

p15INK4B甲基化与骨髓增生异常综合征患者预后及地西他滨疗效的关系

目的 通过检测初诊骨髓增生异常综合征(MDS)患者p15INK4B甲基化水平,观察地西他滨治疗前、后p15INK4B甲基化状态变化,探讨p15INK4B甲基化水平与预后的关系及其对地西他滨疗效的影响.方法 261例初诊MDS患者,其中男143例,女118例,中位年龄52(32 ～78)岁.低危组172例(低危104例、中危-1 68例),高危组89例(中危-2 52例、高危37例).收集患者骨髓单个核细胞,采用甲基化特异性PCR (MSP)检测p15INK4B甲基化水平,按p15INK4B甲基化程度对患者进行生存分析.采用MSP方法分别检测58例MDS患者地西他滨治疗前及治疗2个疗程后骨髓p15INK4B甲基化水平,分析p15INK4B甲基化对地西他滨疗效的影响.结果 低危组患者p15INK4B甲基化水平明显低于高危组患者(117.22对157.63,P＜0.05).p15INK4B甲基化阳性患者2年预期生存(OS)率低于阴性患者(69.8％对91.8％,P＜0.05);在低危组,p15INK4B甲基化阳性患者2年预期OS率低于阴性患者(78.2％对92.0％,P＜0.05);在高危组,p15INK4B甲基化阳性患者OS率及中位OS时间与阴性患者比较差异无统计学意义[35.6％对38.5％,(17.0±9.3)个月对(18.0±5.7)个月,P＞0.05].COX分析结果显示p15INK4B甲基化水平是OS时间的独立预后因素.治疗前p15INK4B甲基化阳性组患者地西他滨治疗的总反应率及完全缓解率与阴性组比较差异无统计学意义(65.9％对76.5％,22.0％对29.4％,P＞0.05).地西他滨治疗有效组p15INK4B甲基化水平在治疗前、后差异无统计学意义(P＞0.05).结论 初诊时p15INK4B甲基化水平高的MDS患者生存时间更短,但p15INK4B甲基化水平对地西他滨疗效无明显影响.     

部分恶性血液病患者红细胞A、H抗原表达与基因启动子CpG岛甲基化研究

目的 研究急性髓系细胞性白血病(AML)与骨髓增生异常综合征(MDS)患者红细胞A、H抗原表达与基因启动子CpG岛甲基化的相关性.方法 应用流式细胞技术检测红细胞表面A、H抗原表达,并应用甲基化特异性PCR(MS-PCR)测定ABO基因启动子CpG岛甲基化.结果 AML与MDS组红细胞表面A抗原阳性表达率和A抗原平均荧...     

p73基因异常表达与急性淋巴细胞白血病相关性的研究

目的探讨急性淋巴细胞白血病(ALL)发生发展的机制。方法应用RT-PCR技术检测42例ALL患者的p73基因mRNA表达情况,并结合患者的临床资料进行分析,同时采用限制性内切酶酶切结合多聚酶链反应方法(REP法)检测p73基因第一外显子甲基化情况。结果42例ALL患者甲基化检测结果28.6%(12/42)存在甲基化,甲基化患者p73mRNA均阴性表达,正常对照组和不存在p73基因甲基化的病人表达p73mRNA。p73mRNA阴性表达与ALL患者首次化疗既获完全缓解及平均缓解时间的降低有明显关系。结论ALL患者p73基因存在较高的阴性表达,p73基因失活在ALL的发生发展中起重要作用。p73基因失活主要原因之一是过度甲基化。p73mRNA检测对判断ALL患者预后有一定的意义。     

地西他滨治疗骨髓增生异常综合征和急性髓系白血病的临床观察

本研究目的是观察地西他滨治疗骨髓增生异常综合征（MDS）和急性髓系白血病（AML）的临床疗效及不良反应。对2009年11月至2011年12月我科收治的应用地西他滨治疗的12例MDS及AML患者进行了回顾性分析。12例中1例MDS-RA,2例MDS-RAEB-Ⅰ,3例MDS-RAEB-Ⅱ,2例AML．M4,2例AML-M5,1例AML-M6,1例AML-M0。8例应用地西他滨化疗5d方案：地西他滨20mg／（m2·d）×5d,4周为1个周期;3例用3d方案：地西他滨15mg／m2,1;L／8h×3d,6周为1个周期;另2例用地西他滨20mg／m2,隔日1次×5次。应用甲基化特异性聚合酶链反应（MS-PCR）检测1例应用地西他滨治疗后达到完全缓解（CR）患者治疗前后ID4基因甲基化表达水平,进一步验证地西他滨的去甲基化作用。结果显示,12例患者中CR2例、部分缓解1例、病情稳定5例、病情进展1例、死亡3例。疾病控制率66．67％（8／12）,有效率25％（3／12）。患者的平均生存期为（11．5±2．1）个月、1年总体生存（OS）率为40％、2年OS率为16．7％。MS-PCR检测显示,地西他滨使ID4基因甲基化水平明显降低。结论：地西他滨能够使MDS患者病情稳定,减轻输血依赖,提高生活质量,甚至能够使部分MDS向白血病转化的患者达到完全缓解。地西他滨能够使ID4基因甲基化水平明显降低。其不良反应主要为骨髓抑制和感染,需输血抗感染等支持治疗,部分患者经积极对症支持治疗可以耐受。本研究中的疗效及生存期与国内研究相类似。     

骨髓增生异常综合征患者ZO-1基因甲基化状态检测的临床意义

本研究探讨紧密连接蛋白（zonula occludens-1,ZO-1）基因异常甲基化在骨髓增生异常综合征（MDS）诊断中基因标志作用的临床意义。采用甲基化特异性-PCR（MS-PCR）方法分析30例健康人骨髓标本及85例MDS患者骨髓标本的ZO-1基因启动子区甲基化状况。结果显示：ZO-1基因在30例健康人标本中均呈完全非甲基化状态;MDS患者骨髓中ZO-1基因甲基化阳性率（56.5%）明显高于健康人,二者差异有统计学意义（p〈0.05）。MDS各亚型病人骨髓ZO-1基因启动子区甲基化阳性率均高于健康人,其差异也具有统计学意义（p〈0.05）。MDS亚型RA、RAS、RAEB、RCMD间ZO-1基因甲基化阳性率差别无统计学意义（p〉0.05）。结论：与健康人相比,MDS病人骨髓中ZO-1基因启动子区呈高甲基化状态并具有特异性。MDS是一种常见的克隆增殖性血液系统疾病,在临床诊断中有时很难与其他疾病相鉴别,而ZO-1基因甲基化模式的改变与MDS的发生密切相关,因此ZO-1基因作为有价值的诊断标志具有重要的临床意义,它可能是一个潜在的血液系统恶性疾病的相关基因。     

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.