IGFBP7基因慢病毒载体的构建及在K562细胞中的表达

本研究构建胰岛素样生长因子结合蛋白7(IGFBP7)基因的慢病毒载体,为研究该基因在白血病中的作用提供基础。采用限制性内切酶酶切获得目的基因,基因重组构建慢病毒载体质粒venus-IGFBP7,用293T细胞包装慢病毒颗粒,感染K562细胞,并采用多种方法鉴定。结果表明,所获IGFBP7基因经测序与GenBank比对序列一致,慢病毒载体质粒venus-IGFBP7经BamHⅠ酶切鉴定片段大小正确,荧光显微镜及流式细胞术检测到绿色荧光蛋白在293T及K562细胞中表达,RT-PCR和Western blot检测到IGFBP7 mRNA和蛋白在K562细胞表达。结论:成功构建带有IGFBP7基因的慢病毒载体,为研究IGFBP7基因在K562细胞中的作用奠定了基础。     

SARI过表达对CBFL细胞凋亡的影响及其机制

目的:探讨SARI基因过表达对核结合因子白血病(CBFL)细胞增殖、凋亡的影响及其机制。方法:通过17号外显子测序验证CBFL细胞模型Kasumi-1细胞负载c-KIT N822K突变。构建SARI基因过表达慢病毒载体,并转染Kasuimi-1细胞。采用荧光定量PCR和Western blot鉴定细胞转染后SARI基因过表达效果。设置空白对照(CON)、空载体对照(NC)和SARI过表达(OE) 3组。MTT法检测各组细胞增殖能力,流式细胞术检测细胞凋亡率,Western blot检测凋亡相关蛋白BCL-2、BAX、Cyto C、Caspase 9、Caspase 3、cleaved-Caspase 3、PARP、cleavedPARP,以及PI3K/Akt通路蛋白PI3K(p85)、p-PI3K(p85)、Akt、p-Akt的表达变化。结果:Kasumi-1细胞具有c-KIT N822K突变。成功构建稳定过表达SARI基因的Kasumi-1细胞。与NC组细胞相比,OE组细胞增殖减缓,凋亡增加;抗凋亡蛋白BCL-2表达降低,促凋亡蛋白BAX表达增高;出现Cyto C蛋白的表达; Caspase 9、Caspase 3表达降低,cleaved-Caspase3表达增高; PARP表达降低,活性剪切体cleaved-PARP表达增高,PI3K/Akt通路蛋白p-PI3K/PI3K、p-Akt/Akt磷酸化水平下调(P 0. 05)。结论:SARI基因过表达可通过线粒体途径诱导CBFL细胞凋亡,机制可能与PI3K/Akt信号通路失活有关。     

蛋白磷酸酶2A催化亚基β(PPP2Cβ)过表达慢病毒载体系统的构建及对K562红系分化的影响

目的:构建蛋白磷酸酶2A的催化亚基PPP2Cβ慢病毒表达载体,获得较高滴度慢病毒颗粒,感染K562细胞建立稳定株,检测过表达PPP2Cβ对K562红系分化的影响。方法:将PPP2Cβ的CDS序列插入到第二代慢病毒表达载体FUGW,并与包装质粒p MD2G和ps PAX2共转染293T细胞,36、48h分两次收集慢病毒上清,应用流式细胞术检测病毒滴度。获得的病毒感染K562细胞,应用Western blot检测过表达效果。采用联苯胺染色以及实时定量PCR检测过表达PPP2Cβ对K562红系分化的影响。结果:成功构建了PPP2Cβ慢病毒表达载体,利用慢病毒系统包装PPP2Cβ的慢病毒,获得了高滴度的慢病毒颗粒,并成功建立了PPP2Cβ的K562细胞稳定株,促进了K562向红系分化。结论:过表达PPP2Cβ能使K562细胞自发的向红系分化。     

下调NPM基因表达对慢性髓系白血病细胞株K562增殖的抑制作用研究

目的:运用RNA干扰技术抑制核仁磷酸蛋白(NPM)的表达,研究NPM基因在慢性髓系白血病细胞株(K562细胞)增殖中的作用及其机制。方法:采用shRNA抑制NPM的表达,应用实时荧光定量PCR技术检测NPM基因的表达,MTS法检测抑制NPM基因对细胞增殖能力的影响,流式细胞术检测细胞周期的改变,Western blot检测细胞周期相关蛋白的表达。结果:成功构建了针对NPM基因的shRNA慢病毒载体,并感染K562细胞。检测结果显示,与对照组相比较,抑制K562细胞NPM基因的表达可以抑制细胞的增殖,减少细胞集落形成。干扰NPM基因可使G0/G1期延长,细胞周期阻滞,这可能与下调NPM基因后激活p21蛋白的表达,进而抑制CDK2/Cyclin E复合物形成有关。结论:下调K562细胞NPM基因的表达,可以通过诱导细胞周期阻滞抑制K562细胞的增殖。     

构建内皮脂肪酶慢病毒表达载体及在HepG2细胞中的表达

背景:内皮脂肪酶主要由血管内皮细胞分泌并作用于血管内皮,内皮脂肪酶可能在动脉粥样硬化等疾病的发生发展中起重要作用,但目前机制尚不明确。目的:构建内皮脂肪酶慢病毒表达载体,产毒感染HepG2细胞,评价重组载体提高内皮脂肪酶表达的效果。方法:RT-PCR调取大鼠内皮脂肪酶基因,SpeⅠ、EcoRⅠ双酶切后,将内皮脂肪酶基因插入慢病毒载体PRRL.sin.CMV.eGFP中构建内皮脂肪酶过表达慢病毒表达载体PRRL.sin.CMV.EL-eGFP;使用PRRL.sin.CMV.EL-eGFP质粒及病毒包装系统于293FT细胞中包装产毒,用于感染HepG2细胞,RT-PCR及Westernblot检测感染后HepG2细胞内皮脂肪酶mRNA及蛋白的表达。结果与结论:酶切及测序鉴定证实内皮脂肪酶基因成功插入PRRL.sin.CMV.EL-eGFP慢病毒表达质粒;转染PRRL.sin.CMV.EL-eGFP慢病毒表达质粒后,HepG2细胞中可见内皮脂肪酶mRNA及蛋白的表达。说明重组PRRL.sin.CMV.EL-eGFP慢病毒可感染HepG2细胞,并使其表达内皮脂肪酶mRNA和蛋白。     

携带人OCT4A基因的慢病毒载体的构建及其稳定转染K562白血病细胞株的建立

本研究旨在构建共表达人OCT4A与绿色荧光蛋白（GFP）慢病毒载体pLVX-OCT4A-ZsGreen1,并通过感染获得稳定表达重组质粒的白血病K562细胞系,观察OCT4A在其中的表达。根据Gen Bank数据库提供的OCT4A mRNA序列,合成编码mRNA基因的特异性引物序列,通过RT-PCR技术扩增出OCT4A全长片段,克隆至pCR-Blunt载体。将OCT4A DNA片段酶切回收后克隆至经EcoR1酶切线性化的慢病毒载体pLVX-IRES-ZsGreen1,构建pLVX-OCT4A-ZsGreen1慢病毒重组质粒,利用测序鉴定所构建的重组载体是否正确。经三质粒包装系统包装后获得高滴度慢病毒颗粒并感染人白血病细胞系K562,通过流式细胞仪分选获得稳定表达的白血病细胞系,应用Real-time PCR和Western blot方法分别检测稳定转染细胞系的OCT4A mRNA和OCT4A蛋白表达。应用CCK-8法和平板细胞克隆形成试验测定OCT4A对K562细胞增殖能力的影响。结果表明,经限制性内切酶检测及基因测序证实成功构建了携带OCT4A基因的重组慢病毒;包装病毒颗粒超速离心后病毒滴度达（1.43±0.25）×10^8U/ml;病毒感染K562细胞后经流式细胞仪分选获得GFP＋细胞,实时定量PCR及Western blot证实病毒感染后OCT4A基因及蛋白的表达可有效上调;CCK-8法及平板集落形成试验显示病毒感染的K562细胞体外增殖活性明显升高,与对照组相比均有统计学差异（P〈0.05）。结论：成功构建了表达OCT4A基因的慢病毒载体,并且获得可稳定上调OCT4A表达的K562细胞株。     

伊马替尼联合P27基因克隆对慢性粒细胞白血病K562细胞的作用

目的:探讨伊马替尼和P27基因克隆联合作用对K562细胞的细胞增殖、细胞周期及凋亡等的调控作用。方法:RT-PCR扩增P27基因,胶纯化回收后连接到T载体测序,序列正确后构建P27-pcDNA3.1载体,将P27-pcDNA3.1与空载体分别以脂质体转染入P27基因缺失的K562细胞,经筛选得到G418抗性的K562细胞株,Westernblot证实转染后有P27蛋白表达,MTT法检测细胞生长指数和细胞存活率,流式细胞仪检测细胞周期和凋亡。结果:表达外源性P27蛋白的P27-pcDNA3.1-K562细胞株生长速度明显慢于对照K562细胞株,流式细胞仪显示G0/G1期细胞增多,S期细胞明显减少,P27-pcDNA3.1-K562细胞与伊马替尼联合应用后凋亡细胞比例明显上升,MTT法显示细胞存活率较P27-K562细胞组及伊马替尼-K562细胞组均明显下降(P<0.01vsP27-K562细胞组,P<0.05vs伊马替尼-K562细胞组)。结论:表达外源P27蛋白联合应用伊马替尼对抑制K562细胞增殖及促凋亡方面具有协同作用。     

构建内皮脂肪酶慢病毒表达载体及在HepG2细胞中的表达

背景：内皮脂肪酶主要由血管内皮细胞分泌并作用于血管内皮,内皮脂肪酶可能在动脉粥样硬化等疾病的发生发展中起重要作用,但目前机制尚不明确。目的：构建内皮脂肪酶慢病毒表达载体,产毒感染HepG2细胞,评价重组载体提高内皮脂肪酶表达的效果。方法：RT-PCR调取大鼠内皮脂肪酶基因,SpeⅠ、EcoRⅠ双酶切后,将内皮脂肪酶基因插入慢病毒载体PRRL.sin.CMV.eGFP中构建内皮脂肪酶过表达慢病毒表达载体PRRL.sin.CMV.EL-eGFP;使用PRRL.sin.CMV.EL-eGFP质粒及病毒包装系统于293FT细胞中包装产毒,用于感染HepG2细胞,RT-PCR及Westernblot检测感染后HepG2细胞内皮脂肪酶mRNA及蛋白的表达。结果与结论：酶切及测序鉴定证实内皮脂肪酶基因成功插入PRRL.sin.CMV.EL-eGFP慢病毒表达质粒;转染PRRL.sin.CMV.EL-eGFP慢病毒表达质粒后,HepG2细胞中可见内皮脂肪酶mRNA及蛋白的表达。说明重组PRRL.sin.CMV.EL-eGFP慢病毒可感染HepG2细胞,并使其表达内皮脂肪酶mRNA和蛋白。     

RNA特异性干扰bcr-abl融合基因联合p27基因克隆对K562细胞的影响

本研究旨在探讨RNA干扰抑制慢性髓系白血病bcr-abl融合基因表达,以及RNAi和p27基因克隆联合作用对K562细胞的细胞增殖、细胞周期及凋亡等的调控作用。以细胞系K562为研究对象,合成并转染针对K562细胞bcr-abl融合基因融合位点的21nt siRNA,应用Northern blot法检测bcr-abl融合基因的表达,Western blot法检测BCR-ABL蛋白及凋亡相关蛋白BCL-xL的表达;同时应用RT-PCR扩增p27基因,构建P27-pcDNA3.1载体,转染p27基因入缺失p27基因的K562细胞,经筛选得到G418抗性的K562细胞株;经Western blot证实有P27蛋白表达后,联合应用RNA干扰及p27基因克隆,通过MTT法及流式细胞仪等检测联合作用后对K562细胞的细胞增殖、细胞周期及凋亡等的调控作用。结果表明,RNA干扰组K562细胞bcr-abl融合基因的表达水平明显下降,转染24小时时有18.4%的K562细胞发生凋亡,细胞凋亡相关蛋白BCL-xL的表达水平下调,出现明显的G1期阻滞;表达外源性P27蛋白的P27-pcDNA3.1-K562细胞株生长速度明显慢于对照K562细胞株。流式细胞仪检测显示,G0/G1期细胞增多,S期细胞明显减少;RNA干扰与p27基因克隆联合作用于K562细胞后,凋亡细胞比例明显上升(33.4%)。MTT法显示,细胞存活率较单纯p27-K562细胞组及RNA干扰-K562细胞组均明显下降(p0.01和p0.05)。结论:特异性siRNA分子可以抑制bcr-abl融合基因的表达,诱导K562细胞分化或凋亡。RNA干扰联合p27基因克隆对抑制K562细胞增殖及促凋亡方面具有协同作用。     

稳定下调血管内皮钙黏蛋白表达对K562细胞药物敏感性的影响

目的:探索稳定下调VE-cadherin蛋白对K562细胞药物敏感性的影响及其可能的机制。方法:设计靶向人VE-cadherin的特异性干扰序列,连同IRES-GFP及NEO基因片段,构建重组慢病毒载体;三质粒系统包装慢病毒颗粒,感染K562细胞,筛选稳定下调VE-cadherin的K562细胞。应用CCK8法测定K562细胞对伊马替尼药物敏感性的变化,AnnexinⅤ/7-AAD标记细胞,流式细胞术检测细胞凋亡;荧光定量PCR检测白血病细胞CD133、ALDH1的转录水平;Western blot方法检测白血病细胞VE-cadherin、BCR-ABL和β-catenin表达水平。结果:成功构建重组慢病毒载体pLB-sh VEC-NEO-IRES-GFP,并包装慢病毒,筛选出稳定下调VE-cadherin蛋白的K562细胞株。稳定下调K562细胞VE-cadherin表达后,在相同药物浓度处理下白血病细胞增殖率降低,凋亡率升高,CD133及ALDH1转录水平降低,BCR-ABL融合蛋白表达水平无明显变化,总β-catenin蛋白表达水平下降,细胞核β-catenin蛋白水平亦下降。结论:稳定下调K562细胞VE-cadherin蛋白的表达可使白血病细胞药物敏感性增高,其机制可能与降低β-catenin蛋白的稳定性、减少β-catenin蛋白的核转位有关的。     

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.