长链非编码RNA ZFAS1在非小细胞肺癌患者组织中表达及作用

目的检测非小细胞肺癌(NSCLC)患者癌组织中锌指结构反义转录本1(zinc finger antisense 1,ZFAS1)表达水平,探讨ZFAS1在NSCLC进展中的生物学作用。方法实时荧光定量RT-PCR检测ZFAS1在NSCLC患者癌组织和癌旁组织中的表达水平。RNA干扰ZFAS1在A549细胞中表达,细胞计数和克隆形成实验检测细胞增殖情况,流式分析检测细胞周期和细胞凋亡,Transwell迁移和基质胶侵袭实验检测细胞迁移和侵袭能力,实时荧光定量RT-PCR检测Cyclin D1、Bcl2、N-cadherin、ZEB1、Slug和Twist基因表达水平变化。结果 ZFAS1在NSCLC患者癌组织中的平均表达水平[0.01(0.002,0.054)]较癌旁组织[0.002(0.001,0.012)]明显升高(Z=-2.638,P0.01)。ZFAS1基因敲减后,A549细胞增殖能力明显减弱(P0.01);A549细胞周期G1期比例升高,S期比例下降(P0.01);A549细胞凋亡比例明显增加(P0.01);A549细胞迁移和侵袭能力明显下降(P均0.01);A549细胞中Cyclin D1、Bcl2、N-cadherin、ZEB1、Slug和Twist基因表达水平均降低(P均0.05)。结论 ZFAS1在NSCLC患者癌组织中呈高表达。ZFAS1基因敲减诱导细胞周期阻滞、凋亡和抑制上皮-间质转变(EMT),减弱NSCLC细胞增殖、迁移和侵袭能力。     

miR-224靶向抑制ECRG4表达对非小细胞肺癌细胞增殖、迁移和侵袭的影响

目的研究微小RNA-224(miR-224)及其靶基因食管癌相关基因4(ECRG4)在非小细胞肺癌(NSCLC)中的表达和对A549细胞增殖、迁移、侵袭的影响。方法利用原位杂交、荧光定量PCR、免疫组织化学和Western blot检测NSCLC临床标本和细胞系中miR-224和ECRG4的表达水平;通过microRNA.org网站预测miR-224与ECRG4的靶向关系,并行双荧光素酶报告基因实验和Western blot进行验证;分析抑制miR-224及联合抑制ECRG4对A549细胞增殖、迁移和侵袭能力的影响。结果 NSCLC癌组织中miR-224表达水平明显高于癌旁组织,ECRG4mRNA和ECRG4蛋白表达水平均明显低于癌旁组织,差异有统计学意义(P0.05)。与健康人肺支气管上皮细胞系HBE相比,NSCLC细胞系L78、A549、H460中miR-224表达水平明显升高,ECRG4mRNA和ECRG4蛋白表达水平明显降低,差异有统计学意义(P0.05)。经microRNA.org网站预测、双荧光素酶报告基因实验和Western blot证实,ECRG4是miR-224的靶基因。抑制miR-224可抑制A549细胞增殖、迁移和侵袭(P0.05),而联合抑制ECRG4则可逆转抑制miR-224对A549细胞的抑制作用(P0.05)。结论 miR-224可通过靶向ECRG4促进NSCLC细胞A549增殖、迁移和侵袭,提示miR-224和ECRG4可能成为诊断和治疗NSCLC的靶点。     

LINC00978在非小细胞肺癌中的临床价值和生物学作用

目的探究长链非编码RNA(long non-coding RNA, lncRNA)LINC00978在非小细胞肺癌(non-small cell lung cancer, NSCLC)中的表达变化及生物学功能,并初步探讨其作用机制。方法采用qRT-PCR检测NSCLC患者肿瘤组织与血清中LINC00978的表达水平。通过CCK-8、平板克隆、Transwell迁移和侵袭实验观察LINC00978敲减和过表达对A549细胞生物学功能的影响。采用流式细胞术、qRT-PCR和western blot探究LINC00978的作用机制。结果 LINC00978在NSCLC患者肿瘤组织(t=2.465,P0.05)和血清(t=8.781,P0.01)中呈高表达。LINC00978敲减抑制A549细胞增殖、迁移和侵袭能力(P均0.01),诱导G_1期阻滞以及细胞凋亡(P均0.01)。LINC00978敲减下调Cyclin D1和Bcl-2的表达而上调Bax的表达(P均0.05)。此外,LINC00978敲减抑制N-cadherin、Vimentin、Snail、Slug和Twist的表达而促进E-cadherin的表达(P均0.05)。LINC00978过表达则具有相反作用。结论 LINC00978在NSCLC中高表达,并促进NSCLC发生、发展,具有成为NSCLC诊断及治疗新靶点的潜能。     

FOXC2通过上调MMP9促进非小细胞肺癌细胞的侵袭迁移

目的:探讨FOXC2对非小细胞肺癌(NSCLC)细胞侵袭迁移的影响。方法:采用实时荧光定量PCR(qRT-PCR)的方法检测人支气管上皮样细胞HBE和人NSCLC细胞A549、H1299中FOXC2的m RNA表达水平。利用pcDNA3.1-FOXC2建立FOXC2过表达的细胞模型,q RT-PCR和蛋白质印迹法(Western blot)验证转染效率。采用Transwell实验检测NSCLC细胞的侵袭迁移能力,qRT-PCR法检测基质金属蛋白酶9(MMP9)的表达水平。结果:人NSCLC细胞H1299、A549中FOXC2的表达水平显著高于人支气管上皮样细胞HBE(P0.05)。过表达FOXC2促进了A549细胞的侵袭迁移,上调了MMP9的表达(P0.05)。结论:NSCLC细胞中FOXC2表达水平明显高于正常支气管上皮样细胞,过表达FOXC2可能通过上调MMP9促进NSCLC细胞的侵袭迁移。     

载脂蛋白M在非小细胞肺癌A549细胞增殖、迁移和侵袭中的作用

目的探讨载脂蛋白M(apo M)对非小细胞肺癌(NSCLC)A549细胞系增殖、迁移、侵袭的影响及与基质金属蛋白酶(MMP)-10的关系。方法以转染apo M慢病毒的A549细胞作为实验组(apo M-OE组),转染空慢病毒的A549细胞作为阴性对照组。分别采用CCK-8法、划痕实验和Transwell侵袭实验检测各组细胞的增殖、迁移和侵袭能力。采用实时荧光定量聚合酶链反应(qRT-PCR)和免疫印迹法检测各组细胞apo M及MMP-10的表达水平。结果与阴性对照组相比,apo M-OE组apo M表达明显上调(P0.01),表达量约为阴性对照组的28倍。apo M-OE组细胞增殖倍数及侵袭率明显高于阴性对照组(P0.001)。与阴性对照组比较,apo M-OE组空白视野显著减少,闭合速率显著加快,即细胞迁移率更大(P0.001)。apo M-OE组中MMP-10 mRNA及apo M、MMP-10蛋白表达量显著高于阴性对照组(P0.05)。结论过表达apo M可增强A549细胞的增殖、迁移和侵袭能力,同时使细胞高表达MMP-10。     

TIGAR调节肺癌细胞的增殖和侵袭能力研究

目的探讨P53下游基因TIGAR在肺癌细胞A549增殖、迁移及侵袭中的作用。方法采用siRNA技术在A549细胞中干扰TIGAR的表达,细胞计数试剂盒(CCK-8)检测细胞增殖,小室法检测细胞迁移,肿瘤细胞侵袭实验检测细胞侵袭,免疫印迹杂交检测相关蛋白水平变化。结果在A549细胞中成功干扰TIGAR后,细胞增殖显著降低(P0.05),细胞迁移和侵袭能力显著减弱,侵袭相关蛋白基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9)的表达量均下调。结论 TIGAR促进肺癌细胞A549的增殖,并促进细胞的迁移和侵袭能力。     

Vinculin表达对非小细胞肺癌A549细胞生物学特征的影响

目的通过体外实验探讨非小细胞肺癌(NSCLC)A549细胞系的Vinculin表达对其生物学特征的影响。方法购买人NSCLC A549细胞系,体外培养,分为对照组、空白载体组以及高表达组三组,高表达组A549细胞系转染Vinculin过表达载体,空白载体组A549细胞系转染空白载体。体外培养48 h后,应用RT-PCR检测各组A549细胞系Vinculin mRNA表达情况,应用MTT法检测细胞活性,Ki-67免疫荧光检测细胞增殖能力,Transwell培养体系Hoechst染色观察A549细胞系迁移侵袭能力。结果 RT-PCR检测结果显示,与对照组、空白载体组比较,高表达组Vinculin mRNA表达量明显增高(P<0.01);MTT法检测结果显示,与对照组、空白载体组比较,高表达组的OD值明显降低(P<0.01);Ki-67免疫荧光检测结果显示,与对照组、空白载体组比较,高表达组Ki-67⁺细胞数量明显减少(P<0.01);Transwell培养体系Hoeschst染色结果显示,与对照组、空白载体组比较,高表达组迁移和侵袭的细胞数量明显减少...     

长链非编码RNA HMMR-AS1促进肺腺癌的恶性进展

目的探讨长链非编码RNA(long noncoding RNA, lncRNA)HMMR-AS1对肺腺癌(lung adenocarcinoma, LUAD)增殖转移的影响。方法采用实时荧光定量聚合酶链反应技术(RT-qPCR)检测LUAD细胞系中HMMR-AS1及其正义链HMMR的表达水平;通过小干扰RNA敲减HMMR-AS1的水平,并利用RT-qPCR检测转染效率及其对HMMR水平的影响;采用CCK-8法、克隆形成实验、细胞凋亡实验、划痕实验、Transwell侵袭实验等表型实验检测干扰HMMR-AS1的表达对A549和H1299细胞生物学功能的影响;western blot检测该两种细胞中HMMR-AS1水平降低对HMMR蛋白表达水平的影响。结果与正常肺上皮细胞BEAS-2A相比,LUAD细胞系A549和H1299中HMMR-AS1的表达水平分别上调了3.06倍和5.02倍(P0.05);转染小干扰RNA后A549和H1299细胞中HMMR-AS1的表达水平明显下降(P0.05),并且明显抑制HMMR的转录和蛋白质表达水平(P0.05);表型实验结果显示,与阴性对照组比较,敲减HMMR-AS1能够抑制LUAD细胞的生长、迁移和侵袭能力,并促进凋亡。结论 LncRNA HMMR-AS1能够促进LUAD细胞的生长、迁移和侵袭能力,影响肺腺癌的恶性进展。     

GREM1对胃癌细胞增殖迁移的作用

目的检测胃癌细胞中GREM1基因的表达,探讨其对胃癌细胞生物学特性的影响并评估其在胃癌诊断和胃癌患者预后中的临床价值。方法运用数据库分析GREM1基因在胃癌组织和癌旁组织中的表达差异,评估GREM1基因表达水平对胃癌患者预后的相关性。Western blot检测胃癌细胞系中GREM1蛋白质表达水平。AGS细胞中沉默GREM1基因后,采用平板克隆、Transwell和western blot检测其对胃癌细胞增殖、迁移、上皮间质转化(EMT)发生及Wnt/β-catenint通路的影响。结果 Kaplan-Meier分析表明,GREM1基因高表达患者总体生存率(OS)和无进展生存率(PFS)均降低。GREM1蛋白在AGS细胞系中的表达水平(1.967±0.056)最高。平板克隆、Transwell及western blot实验显示,沉默GREM1基因可导致胃癌细胞的增殖及迁移能力降低(t分别为22.00,29.60;P均0.01);E-cadherin表达上升(t=10.65,P0.01),ZEB1、MMP2表达均下降(t分别为10.74和13.67,P均0.01);Wnt/β-catenin通路中β-catenin、CyclinD1、c-myc、p-GSK3β和PCNA的表达水平均降低(t分别为12.65,16.21,8.74,7.75和8.42;P均0.01)。结论 GREM1通过激活Wnt/β-catenin诱导EMT发生,促进肿瘤转移和生长。GREM1可作为新的胃癌分子诊断和预后指标。     

沉默Sema6A基因对骨肉瘤细胞U-2OS增殖和转移的影响

目的检测信号素6A(Sema6A)基因在骨肉瘤细胞系中的表达情况,探讨RNA干扰Sema6A基因对骨肉瘤U-2OS细胞增殖、侵袭及迁移的影响。方法实时定量逆转录聚合酶链反应(qRT-PCR)检测骨肉瘤细胞株U-2OS、Saos-2及MG-63中Sema6A的mRNA表达水平。设计并合成针对Sema6A的小干扰RNA(siRNA)3条及阴性对照siRNA,在Lipofectamine RNAi MAX介导下转染U-2OS细胞,通过qRT-PCR检测Sema6A mRNA水平表达及蛋白质印迹法检测Sema6A蛋白水平,进而筛选出转染效率高的siRNA,CCK-8检测各组细胞活力,Transwell细胞侵袭和迁移实验检测各组细胞的侵袭及迁移能力。结果骨肉瘤细胞株中,Sema6A mRNA在U-2OS中表达最高。瞬时转染Sema6A siRNA的U-2OS细胞中Sema6A的mRNA及蛋白水平较阴性对照组均下降,与阴性对照组相比,实验组细胞的增殖、侵袭及迁移能力均增强,差异有统计学意义(P0.05)。结论 RNA干扰沉默Sema6A基因可以增强U-2OS细胞的增殖、侵袭及迁移能力。     

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.