黄芪甲苷通过抑制AKT和NF-κB通路诱导胃癌MGC-803细胞凋亡

目的观察黄芪甲苷对胃癌MGC-803细胞凋亡的影响并探讨其诱导凋亡可能的机制。方法应用四唑盐比色法(MTT法)观察黄芪甲苷对胃癌MGC-803细胞增殖的影响,应用流式细胞术检测其对胃癌MGC-803细胞周期和凋亡的影响,应用蛋白质免疫印迹(Western blot)方法观察其对蛋白激酶B(AKT)和核因子κB(NF-κB)等相关信号通路蛋白及凋亡相关蛋白B淋巴细胞瘤-2(BCL-2)、BCL2-Associated X的蛋白质(BAX)、天冬氨酸蛋白水解酶3(Caspase-3)的影响。结果黄芪甲苷能够抑制胃癌MGC-803细胞的增殖并促进其凋亡,并呈时间和剂量依赖性。同时其能够抑制AKT及NF-κB通路,抑制抗凋亡蛋白BCL-2的表达,促进促凋亡蛋白BAX的表达,降低BCL-2/BAX的比例,增加Caspase-3的活化。结论黄芪甲苷能够促进胃癌MGC-803细胞凋亡,推测可能通过抑制AKT和NF-κB通路,降低BCL-2/BAX的比例,增加Caspase-3的活化的机制而实现。     

miR-193a-3p在胃癌中调节细胞凋亡的分子机制研究

目的探讨微小RNA 193-a-3p(miR-193a-3p)在胃癌中调节细胞凋亡的分子机制。方法运用实时荧光定量PCR在健康人胃黏膜细胞系GES-1、人胃癌细胞系MKN-45、SGC-7901、MGC-803、SNU16检测miR-193a-3p的相对表达水平;通过TargetScan在线分析和荧光素酶报告系统检测miR-193a-3p的靶基因;过表达或敲低miR-193a-3p后,免疫印迹试验检测靶基因和细胞凋亡标志蛋白的表达量以及细胞凋亡检测试验(Annexin V双染色法)检测细胞的凋亡水平。结果相比于健康人胃黏膜细胞系GES-1,miR-193a-3p在人胃癌细胞系MKN-45、SGC-7901、MGC-803、SNU16中低表达(P0.05),并且MGC-803的表达量最低。miR-193a-3p靶向P21活化激酶4(PAK4)的3端非编码区。过表达miR-193a-3p后,PAK4的表达量降低,而凋亡标志蛋白cleaved-caspase3表达量升高,胃癌细胞MGC-803的凋亡水平上升(P0.05)。敲低miR-193a-3p后,PAK4的表达量升高,而凋亡标志蛋白cleaved-caspase3表达量降低,胃癌细胞MGC-803的凋亡水平下降(P0.05)。结论 miR-193a-3p通过靶向调节PAK4促进胃癌细胞发生细胞凋亡。     

γ-干扰素对MGC-803胃癌细胞株诱导凋亡作用及其机制研究

目的　研究干扰素(Interferon-γ,IFN-γ)对MGC-803胃癌细胞株的抗增殖和诱导凋亡作用,并 探讨其诱导凋亡作用的机制。方法　应用MTT法检测IFN对MGC-803胃癌细胞株的抗增殖作用的影响及细 胞毒活性;通过MTT染色法、HE染色、扫描电镜观察凋亡细胞的形态学改变;应用(TdT-mediateddUTPnick endlabeling)TUNEL法定量检测IFN γ诱导凋亡的情况;通过免疫细胞化学法测定P53、P16、C-myc、Bcl-2、Bax 的阳性表达率。结果　IFN-γ在1250～10000u/ml范围内对MGC-803胃癌细胞均有抑制作用(P<0.05), MTT染色法观察:正常肿瘤细胞着色均匀,凋亡细胞缩小成深蓝色,死亡细胞空泡样,不着色。扫描电镜及 TUNEL法均观察到实验组细胞体积变小,可见凋亡小体突出于肿瘤细胞表面。凋亡率随作用时间和浓度增加而 升高,实验组P53、P16、C-myc蛋白表达没有明显变化(P>0.05),Bcl-2蛋白表达下降,Bax蛋白表达上调,与对 照组相比均有显著性差异(P<0.01)。结论　IFN-γ可抑制MGC-803胃癌细胞增殖,诱导肿瘤细胞凋亡。其 作用机制可能是与通过上调肿瘤细胞Bax蛋白表达,下调Bcl-2蛋白表达有关。     

Cdx2-siRNA对胃癌细胞Survivin和Caspase-9表达的影响

目的:观察Cdx2小分子干扰RNA(Cdx2-siRNA)对人胃癌MGC-803细胞凋亡及Survivin、Caspase-9表达的影响。方法:设计合成针对Cdx2基因的siRNA寡核苷酸以及阴性对照siRNA。将人胃癌MGC-803细胞接种于培养板中,分为3组,正常对照组不作处理,阴性对照组和Cdx2-siRNA组分别用LipofectamineTM2000将阴性对照siRNA或Cdx2-siRNA成功转染进细胞内。应用流式细胞仪检测转染24h时MGC-803细胞凋亡的变化,并应用RT-PCR和Western blot技术检测胃癌MGC-803细胞中Survivin和Caspase-9基因mRNA和蛋白的表达。结果:Cdx2-siRNA组MGC-803细胞的凋亡率为(12.5±0.6)%,明显高于阴性对照组和正常对照组(P<0.05);Cdx2-siRNA组Survivin mRNA和蛋白的表达量分别为(0.14±0.04)和(0.51±0.12),明显低于阴性对照组和正常对照组(P<0.05);Cdx2-siRNA组Caspase-9 mRNA和蛋白的表达量分别为(0.98±0.24)和(1.78±0.41),明显高于阴性对照组和正常对照组(P<0.05);而阴性对照组和正常对照组比较差异无统计学意义(P>0.05)。结论:Cdx2-siRNA促进人胃癌细胞的凋亡,其机制可能与其抑制Survivin的表达、增强Caspase-9的活性有关。     

熊果酸对人胃癌细胞株MGC-803增殖的影响及机制

目的 观察熊果酸(Ursolic acid, UA)对胃癌细胞株(MGC-803)的抑制增殖作用。方法 培养胃癌细胞株(MGC-803)，以MTT比色法及克隆形成实验测定UA对胃癌细胞株MGC-803抑制增殖的作用。免疫印记法测定p-ERK1/2、Cyclin D1及p21waf/cip1表达。结果 MTT实验及克隆分析法均显示UA明显抑制MGC-803细胞增殖，免疫印记法显示UA抑制p-ERK1/2、Cyclin D1表达，同时上调p21waf/cip1表达。结论 熊果酸可以抑制胃癌细胞株(MGC-803)增殖，机制与影响p-ERK1/2及下游Cyclin D1、p21waf/cip1表达有关。     

高容量血液滤过对多器官功能障碍综合征犬内质网应激标志蛋白Caspase-12基因水平及表达的影响

目的探讨高容量血液滤过(HVHF)治疗多脏器功能障碍综合征(multiple organ dysfunction syndrome,MODS)对内质网应激相关性细胞凋亡及应激标志蛋白Caspase-12基因水平及表达的影响,并探讨高容量血液滤过(high volume hemofiltration,HVHF)治疗MODS的机制。方法经二次打击建立犬MODS模型。分HVHF组和MODS组,于术前(T1)、内毒素注射完后0h(T2)、6h(T3)、12h(T4)及24h(T5)留血标本。体内实验:应用荧光定量PCR法检测肝、肾及肺组织Caspase-12m RNA基因表达水平。体外实验:2组血清体外诱导人脐静脉内皮细胞(HUVECs)建立内质网应激凋亡模型。测定si RNA干扰前后Caspase-12m RNA基因表达、蛋白表达及内皮细胞凋亡率。结果体内实验:与MODS组相比,HVHF组肾、肺组织Caspase-12m RNA基因表达水平明显降低,差异有统计学意义(P0.05),而肝组织Caspase-12m RNA基因表达水平无明显差异(P0.05)。体外实验:干扰前与MODS组相比,HVHF组在T2-T5HUVECs Caspase-12m RNA基因表达水平、Caspase-12蛋白表达水平及细胞凋亡率均明显降低,差异有统计学意义(P0.01)。干扰后2组HUVECs Caspase-12m RNA基因表达水平、Caspase-12蛋白表达水平及细胞凋亡率均较干扰前显著降低,差异有统计学意义(P0.01)。结论内质网应激凋亡信号通路在MODS演进过程中占重要地位,经过HVHF治疗后内质网应激(ERS)凋亡信号通路中的关键蛋白Caspase-12基因表达、Caspase-12蛋白表达水平及细胞凋亡率均明显下降,此作用有助于减轻机体细胞凋亡程度,有效地救治MODS。而RNA干扰(RNAi)技术有望成为治疗MODS新的治疗靶点。     

曲古霉素A诱导胃癌细胞系BGC-803凋亡的机制研究

目的利用曲古霉素A(TSA)处理人胃癌细胞系BGC-803后观察细胞增殖情况并研究了凋亡相关基因表达量的变化。方法本研究通过TSA处理胃癌细胞系BGC-803细胞后,利用MTT、Real-time PCR、Western blot等试验方法,采用SPSS 17.0统计软件进行数据分析。结果 75 ng/m L的TSA处理48 h相较其他试验组,可显著抑制胃癌细胞系BGC-803细胞增殖(P0.05),且其抑制作用具有剂量、时间依赖性;使促凋亡蛋白BAX、Caspase-3的mRNA、蛋白表达量升高,从而发挥促进细胞凋亡的作用。结论 TSA诱导BGC-803调亡是通过Caspase依赖途径进行的,且随着时间和剂量的增加,TSA对胃癌细胞的抑制效果越来越显著。     

姜黄素对人类绒毛膜癌细胞增殖、凋亡的影响及机制

目的探讨姜黄素对人类绒毛膜癌细胞增殖、凋亡的影响及机制。方法将人类绒毛膜癌细胞JEG-3细胞株随机分为对照组和姜黄素干预组。姜黄素干预组加入100μl不同浓度的姜黄素与细胞孵育24 h;对照组只加入PRMI 1640培养液与等体积的二甲基亚砜(DMSO),MTT检测检测细胞抑制率。姜黄素干预组加入100μl姜黄素(终浓度为40μmol/L)与细胞孵育24 h后,TUNEL法检测细胞凋亡情况,Western blot检测增殖相关蛋白Ki-67和细胞周期蛋白D1(Cyclin D1)的表达,以及凋亡相关蛋白半胱氨酸蛋白酶(Caspase-3)、B淋巴细胞瘤-2(Bcl2)和Bcl2相关X蛋白(Bax)的表达。结果与对照组相比,姜黄素干预组JEG-3细胞培养24 h后吸光度显著下降(P 0. 05);姜黄素对JEG-3细胞抑制作用呈药物浓度依赖性,半抑制浓度(IC50)为(39. 33±1. 02)μmol/L;与对照组相比,JEG-3细胞的凋亡率明显升高,增殖相关蛋白Ki-67和Cyclin D1的表达显著降低(P 0. 05),凋亡相关蛋白Caspase-3和Bax的蛋白表达量显著升高(P 0. 05),Bcl-2的表达量显著降低(P 0. 05)。结论姜黄素可以下调人类绒毛膜癌细胞增殖相关蛋白的表达,上调凋亡相关蛋白的表达,调控其增殖和凋亡。     

沙樱桃脂类对人MGC-803细胞抑制作用的实验研究

目的探讨沙樱桃脂类对人胃癌MGC-803细胞的毒性作用及其机制。方法采用台盼蓝法检测细胞数、流式细胞仪检测细胞凋亡。将人MGC-803细胞分别与不同浓度的沙樱桃脂类(沙樱桃脂组)、5-FU(5-FU组)及PBS溶液(对照组)共同培养,观察沙樱桃脂类对人胃癌MGC-803细胞增殖的抑制作用。结果对人MGC-803细胞分别给予0.5、1.0、3.0mg/L沙樱桃脂类及10mg/L5-FU,培养6h、12h、18h、24h和48h,活细胞数随沙樱桃脂类浓度的增加和作用时间的延长而减少,与5-FU组相似(P>0.05),而与对照组比,差异有统计学意义(P<0.01);分析存活细胞率,表明人MGC-803细胞增殖被阻滞于G0G1期。在给予沙樱桃脂类0.5、1.0、3.0mg/L和5-FU培养48h后,人MGC-803细胞的凋亡率分别为13.8%、19.6%、26.2%和21.1%。结论沙樱桃脂类对人胃癌MGC-803细胞具有显著毒性作用,其机制可能与沙樱桃脂类可抑制MGC-803细胞分裂,并诱导其凋亡有关。     

HMGB1对膀胱癌细胞增殖和凋亡的影响及机制

目的研究高迁移率蛋白1(HMGB1)对膀胱癌细胞增殖、凋亡的影响及机制。方法膀胱癌细胞BIU-87分成对照组、siRNA control组、HMGB1 siRNA组共3组,siRNA control组、HMGB1 siRNA组细胞为感染siRNA control慢病毒和HMGB1 siRNA慢病毒的BIU-87细胞,对照组为不感染慢病毒的BIU-87细胞。用Realtime PCR和Western blot检测细胞中HMGB1表达水平。MTT检测膀胱癌细胞增殖活性,流式细胞术检测膀胱癌细胞凋亡,Western blot检测膀胱癌细胞中CCAAT/增强子结合蛋白同源蛋白(CHOP)、活化型Caspase-3(Cleaved Caspase-3)、活化型Caspase-12(Cleaved Caspase-12)、活化转录因子4(ATF4)蛋白水平。结果 HMGB1 siRNA可以下调膀胱癌细胞中HMGB1的表达和转录,siRNA control对膀胱癌细胞中HMGB1表达没有影响。沉默HMGB1后膀胱癌细胞增殖能力下降,细胞凋亡率升高,细胞中凋亡蛋白Cleaved Caspase-3、Cleaved Caspase-12表达水平升高,内质网应激蛋白CHOP、ATF4表达水平也升高。结论 HMGB1沉默可以通过内质网途径诱导膀胱癌细胞凋亡,抑制膀胱癌细胞增殖。     

Guns and Knives in New Mexico: Patterns of Penetrating Trauma, 1978–1993

Objective: To identify patterns of nonfatal and fatal penetrating trauma among children and adults in New Mexico using ED and medical examiner data.
Methods: The authors retrospectively sampled in 5-year intervals all victims of penetrating trauma who presented to either the state Level-1 trauma center or the state medical examiner from a 16-year period (1978–1993). Rates of nonfatal and fatal firearm and stabbing injury were compared for children and adults.
Results: Rates of nonfatal injury were similar (firearm, 34.3 per 100,000 person-years; stabbing, 35.1). However, rates of fatal injury were significantly different (firearm, 21.9; stabbing, 2.7; relative risk: 8.2; 95% confidence interval: 5.4, 12.5). From 1978 to 1993, nonfatal injury rates increased for children (p = 0.0043) and adults (p < 0.0001), while fatal penetrating injury remained constant. The increase in nonfatal injury in children resulted from increased firearm injury rates. In adults, both stabbing and firearm nonfatal injury rates increased.
Conclusions: Nonfatal injury data suggest that nonfatal violence has increased; fatal injury data suggest that violent death rates have remained constant. Injury patterns vary by age, mechanism of trauma, and data source. These results suggest that ED and medical examiner data differ and that both are needed to guide injury prevention programs.     

Interprofessional approaches to creating safe,high quality health care

Three supplementary perspectives are presented arguing that interprofessional collaboration is both necessary and desirable. Nonetheless, there are often too many serious intra-professional barriers and obstacles to interprofessional collaboration to make it successful. Some of these barriers, it is argued and illustrated, are found in the multiple ways in which professional identity is tacitly acquired and embodied in the practitioners' habitual, everyday practice. The paper then explores ways in which reflection, especially Second order reflection, can help to elucidate and overcome these obstacles, as well as increasing professional adaptability and competence.     

Systematic Reviews Published in the January 2011 Issue of The Cochrane Library

ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly as a DVD and monthly online. The January 2011 issue (first quarterly DVD for 2011) contains 4515 complete reviews, 1985 protocols for reviews in production, and 13,521 one-page summaries of systematic reviews published in the general medical literature. In addition, there are citations of 641,000 randomized controlled trials, and 14,018 cited papers in the Cochrane methodology register. The health technology assessment database contains over 9300 citations. One hundred and seven new reviews have been published in the last 3 months, of which five have potential relevance for practitioners in pain and palliative medicine.     

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.     

Prise en charge des entorses externes de cheville : étude clinique préliminaire sur l’efficacité du dispositif Myolux™

Ankle sprains are the most common injury of the musculoskeletal system and are associated with significant societal and economic impacts. It has been proven that classical therapeutic strategies may not be effective in preventing recurrent injuries: the recurrence rates reported in the literature can reach 73%. In order to provide an effective rehabilitation solution, a destabilizing orthosis was developed. This device is equipped with a mechanical articulator reproducing the subtalar mechanics and placed under the heel. In this paper, we present the main results of a preliminary clinical study conducted between 2004 and 2007. All subjects included in this study were treated with the abovementioned orthosis during 10 rehabilitation sessions of 30 minutes each. Data show a relatively low recurrence rate of 12% for the overall population. Moreover, it's of primary importance to note that this satisfactory ratio is largely reduced (3% of recurrence rate) for the 29 patients who performed one training session per month after the 10th initial rehabilitation sessions. Hence, the destabilizing orthosis appears to be an effective solution to prevent recurrent ankle sprains. However, joint protection requires long-term and regular training sessions. This result has motivated the development of a similar device allowing patients to perform training sessions at home. Finally, data obtained in this study are promising awaiting the final results of the comparative, multicentric and independent clinical trials currently managed by the Hospices Civils de Lyon.