线粒体分裂蛋白抑制剂对β淀粉样蛋白诱导小胶质细胞凋亡的作用及机制

目的:研究线粒体分裂蛋白抑制剂对β淀粉样蛋白(Aβ)诱导小胶质细胞凋亡的作用及其机制。方法:随机将BV-2小胶质细胞分为con组、Aβ组、mdi组和Aβ+mdi组,con组不做特殊处理,mdi组培养基中加入10μmol/L mdivi-1,Aβ组培养基中加入20μmol/L Aβ,Aβ+mdi组培养基分别加入2、5、10、20μmol/L mdivi-1和20μmol/L Aβ。采用MTT法检测细胞存活率,TUNEL染色检测细胞凋亡,Western blot法检测Drp1、CytC和Caspase-3蛋白水平变化,RT-PCR法检测CA11b mRNA表达变化。结果:与con组相比,Aβ组的细胞存活率明显下降,凋亡显著增加,CA11b mRNA上升,线粒体Drp1、细胞浆CytC和激活的Caspase-3增加,差异有统计学意义(P<0.05);与Aβ组相比,Aβ+mdi组的细胞存活率明显上升,凋亡显著减少,CA11b mRNA下降,线粒体Drp1、细胞浆CytC和激活的Caspase-3减少,差异有统计学意义(P<0.05)。结论:线粒体分裂蛋白抑制剂对Aβ诱导小胶质细胞凋亡有保护作用,其机制可能为抑制线粒体/CytC/Caspase-3凋亡途径。     

银杏提取物对淀粉样β蛋白致PC12细胞凋亡的保护作用

目的:探讨银杏提取物对淀粉样β蛋白(Aβ)致PC12细胞凋亡的保护作用。方法:应用二甲基噻唑二苯基四唑溴盐(MTT)分析法研究细胞活性的改变,应用激光共聚焦显微镜进行线粒体膜电位测定。TUNEL法测定细胞凋亡率。结果:随着Aβ浓度的加大,PC12细胞的存活率逐渐降低。同时给予Aβ及不同剂量的银杏提取物,可明显提高PC12细胞的存活率。银杏提取物可减少Aβ所致的细胞凋亡,Aβ组细胞凋亡率为63%,高于银杏组21%(χ2=36.2069,P=0.0000)。结论:银杏提取物可抑制Aβ诱导的PC12细胞凋亡。     

隐丹参酮与多奈哌齐合用对Aβ诱导大鼠皮层神经元损伤的保护作用

目的：观察隐丹参酮与多奈哌齐合用对Aβ42诱导大鼠皮层神经元凋亡的保护作用及其可能机制。方法通过Aβ42诱导体外培养的大鼠皮层神经元,建立阿尔茨海默病（Alzheimer＆#39;s disease,AD）细胞模型,MTT比色法检测细胞存活率,流式细胞仪检测细胞凋亡率,分光光度法测定SOD活性、MDA含量及LDH漏出量。结果成功建立大鼠皮层神经元AD细胞模型,经检测模型组MDA含量及LDH漏出量显著高于空白对照组且SOD活力明显低于空白对照组;隐丹参酮组、隐丹参酮加多奈哌齐组MDA含量及LDH漏出量均低于模型组,且SOD活力明显高于模型组,差异均有统计学意义（P〈0.05）;隐丹参酮与多奈哌齐合用在抑制神经元凋亡,降低MDA含量、LDH漏出量及提高SOD活性方面的效果显著强于隐丹参酮与多奈哌齐单用。结论隐丹参酮与多奈哌齐合用对Aβ损伤细胞有保护作用,其机制可能与协同抗氧化能力有关。     

银杏提取物对淀粉样β蛋白致PC12细胞凋亡的保护作用

目的：探讨银杏提取物对淀粉样β蛋白(Aβ)致PC12细胞凋亡的保护作用。方法：应用二甲基噻唑二苯基四唑溴盐(MTT)分析法研究细胞活性的改变，应用激光共聚焦显微镜进行线粒体膜电位测定。TUNEL法测定细胞凋亡率。结果：随着Aβ浓度的加大，PC12细胞的存活率逐渐降低。同时给予Aβ及不同剂量的银杏提取物，可明显提高PC12细胞的存活率。银杏提取物可减少Aβ所致的细胞凋亡，Aβ组细胞凋亡率为63％，高于银杏组21％(x^2=36．2069，P=0．0000)。结论：银杏提取物可抑制Aβ诱导的PC12细胞凋亡。     

何首乌苷对奥沙利铂诱导大鼠星形胶质细胞凋亡的影响

目的探讨何首乌苷保护大鼠星形胶质细胞免于奥沙利铂诱导凋亡的作用及可能机制。方法原代培养大鼠星形胶质细胞,并利用流式检测所培养细胞胶质纤维酸性蛋白的表达情况。利用10μmol/L奥沙利铂处理胶质细胞24 h建立凋亡模型,加入浓度梯度的何首乌苷预处理胶质细胞24 h后与奥沙利铂共同作用24 h,台盼蓝染色法观察细胞存活。随后通过Annexin V-FITC/PI双染法流式检测各组凋亡率和检测Caspase-3的活性水平。进而利用JC-1染色法流式检测细胞线粒体膜电位,并利用实时定量RT-PCR与免疫印迹检测BAX与BCL2的m RNA水平和蛋白表达量。结果培养的大鼠星形胶质细胞表达胶质纤维酸性蛋白率达95%以上。奥沙利铂处理诱导凋亡24 h后,对照组胶质细胞凋亡率及Caspase-3活性上升并降低线粒体膜电位,加何首乌苷处理后凋亡率及Caspase-3活性下降而线粒体膜电位上升。相对于对照组,加入何首乌苷处理后BAX基因的m RNA及蛋白表达水平降低,BCL2基因的m RNA及蛋白表达水平上升。结论何首乌苷能恢复凋亡相关基因BAX与BCL2的平衡,保护大鼠皮质星形胶质细胞的线粒体膜电位,并使胶质细胞免于奥沙利铂诱导的凋亡作用。     

急变期慢性髓系白血病骨髓间充质干细胞下调白血病细胞凋亡

本研究旨在观察慢性髓系白血病（CML）骨髓间充质干细胞（BMMSC）对K562细胞和原代急变期CML白血病细胞增殖和凋亡的影响及其可能的机制及意义.K562细胞和原代急变期CML白血病细胞分别与不同组别的BMMSC共培养,应用MTT法检测细胞增殖,流式细胞术检测细胞凋亡及细胞线粒体膜电位,Western blot检测凋亡相关蛋白caspase-8、caspase-9、激活的caspase-3的表达.结果表明：CML急变期BMMSC不能抑制原代CML急变期白血病细胞的增殖,对K562仅有轻度的抑制作用;CML急变期BMMSC提高阿霉素处理后的K562细胞的存活率,并保护K562及原代CML-Bp骨髓单个核细胞,抑制阿霉素诱导的细胞凋亡（P＜0.05）;与CML慢性期及正常组BMMSC相比较,CML急变期BMMSC对白血病细胞的保护作用最强,其细胞共培养组的细胞线粒体膜电位下降最少（P＜0.05）;检测CML急变期BMMSC共培养组的K562显示,活性Caspase-3表达均较单独K562＋ ADM组明显下调,caspase-9表达显著增加（P＜0.05）.结论：CML急变期BMMSC下调阿霉素诱导的白血病细胞的凋亡,其机制可能与抑制细胞线粒体膜电位的下降,稳定caspase-9蛋白非活性的表达和下调caspase-3蛋白的激活有关.     

血小板生成素通过PI3K/AKT通路防止CoCl_2诱导内皮细胞凋亡

目的:研究血小板生成素(TPO)对化学性缺氧模拟剂氯化钴(CoCl_2)诱导的人脐静脉内皮细胞(HUVEC)凋亡的作用及其机制。方法:实验分为4组,体外培养的未经处理的HUVEC为对照组,CoCl_2与HUVEC共培养的为CoCl_2组,在加入CoCl_2前用TPO预处理HUVEC为TPO+CoCl_2组,TPO单独处理HUVEC为TPO组。使用CCK-8法检测各组细胞的细胞活性,流式细胞术检测细胞凋亡率、凋亡蛋白酶Caspase-3的表达以及线粒体膜电位的变化。Western blot检测TPO对AKT通路磷酸化水平表达的影响。结果:CoCl_2可以明显抑制HUVEC的生长,细胞存活率随着CoCl_2的浓度的升高逐渐下降(r=-0.997);与对照组相比较,CoCl_2组的细胞凋亡率明显升高(P0.001),而TPO+CoCl_2组的细胞凋亡率较前者明显下降(P0.001),提示TPO对HUVEC有保护作用。TPO还能减少细胞凋亡蛋白酶Caspase-3的表达和抑制细胞线粒体膜电位的降低,以及刺激HUVEC PI3K/AKT通路的活化。结论:TPO具有防止化学性缺氧所致的细胞凋亡作用,其机制可能是通过活化PI3K/AKT通路,从而减少细胞凋亡蛋白酶Caspase-3的活化表达和稳定线粒体膜电位来发挥细胞保护作用。     

华蟾素抗肺癌细胞的实验研究

目的探究华蟾素的抗肺癌活性。方法培养肺癌A549细胞,对华蟾素、环孢素-A(CsA)、萨菲菌素A(SfA)作用于肺癌A549细胞后的细胞存活率、细胞死亡率、细胞凋亡率以及线粒体膜电位(MMP)进行检测;建立小鼠移植瘤模型,对华蟾素作用于小鼠移植瘤模型后的肿瘤体积以及小鼠存活率进行检测,并整理数据进行统计学对比分析。结果通过小鼠移植瘤模型,该研究发现华蟾素抑制了A549肺癌细胞在体内生长。研究结果表明亲环蛋白D(Cyp-D)依赖的线粒体通透性转换孔(mPTP)开放可调节由华蟾素诱导的肺癌细胞的非凋亡性死亡。Cyp-D的抑制剂环孢素-A(CsA),mPTP的阻断剂萨菲菌素A(SfA)极明显地阻碍了由华蟾素诱导的线粒体膜电位(MMP)的下降和A549细胞的死亡(非凋亡)。结论华蟾素可以诱导肺癌细胞死亡,值得进一步研究,有望成为一种新型的抗肺癌药物。     

硫氧还蛋白对大鼠脊髓损伤的保护作用

目的:观察硫氧还蛋白(Trx)对大鼠脊髓损伤(SCI)后脊髓组织丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性及细胞凋亡的影响,探讨其对SCI的保护作用。方法:成年Wister大鼠90只,随机分为假手术组(Sham组)、单纯SCI组(A组)及Trx治疗组(B组),A、B两组采用改良Allen法制成大鼠SCI模型,术后立即分别腹腔注射等体积生理盐水、Trx(0.75mg/kg),再于术后2h、8h、1d、2d、7d、14d取损伤处脊髓标本,测定伤段脊髓组织MDA、SOD含量及凋亡细胞数。结果:A组大鼠MDA伤后8h开始升高,1d达高峰,2d开始下降,SOD伤后8h开始下降,1d达低谷,2d开始升高,两者与Sham组3个时间点相比,差异均具有显著性(P<0.05或P<0.01)。B组大鼠MDA伤后8h、1d及2d与A组相比明显减少,SOD伤后8h、1d及2d与A组相比明显升高,两者差异均具有显著性(P<0.05或P<0.01)。A组凋亡细胞百分率伤后1d开始升高,2d达高峰,7d开始下降。B组凋亡细胞百分率伤后1d、2d及7d与A组相比明显减少,差异具有显著性(P<0.05或P<0.01)。结论:SCI后早期应用Trx可以抑制MDA上升,提高SOD含量,减少细胞凋亡,对脊髓损伤有保护作用。     

TLR4受体拮抗剂对Aβ1-42诱导的小胶质细胞炎性因子分泌的影响

目的探讨体外培养条件下Toll样受体4(TLR4)拮抗剂对β淀粉样蛋白1-42片段(Aβ1-42)诱导的小胶质细胞炎性因子分泌的影响。方法用不同浓度的Aβ1-42(终浓度为0、1、5、10、20、40、80μmol/L)及TAK-242(终浓度1μmol/L)处理小鼠小胶质细胞(BV2),24 h后取其上清干预小鼠海马神经元(HT22)。48 h后,应用CCK8方法检测HT22细胞的存活,ELISA方法检测BV2细胞上清液中TNF-α、IL-1β水平。结果与空白对照组相比,Aβ1-42直接作用于体外培养的小胶质细胞后,以剂量依赖的方式诱导TNF-α、IL-1分泌的增加(P0.05);与空白对照组相比,Aβ1-42诱导组培养液干预的HT22细胞存活率显著下降(P0.05)。给予TLR4拮抗剂TAK-242干预后,Aβ1-42诱导TNF-α、IL-1分泌的作用被抑制,细胞分泌TNF-α、IL-1β水平较Aβ1-42诱导组显著降低(P0.05),HT22细胞存活率也显著提高(P0.05)。结论阻断TLR4可抑制Aβ1-42诱导的小胶质细胞炎性因子的分泌,并减轻活化的小胶质细胞对海马神经元的损害。     

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.     

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.     

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.     

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.     

Clinical Trial Updates and Hotline Sessions presented at the European Society of Cardiology Congress 2007

This article provides information and commentaries on trials which were presented at the Hotline and Clinical Trial Update Sessions at the European Society of Cardiology Congress 2007 in Vienna. The key presentations were performed by leading experts in the field with relevant positions in the trials or registries. It is important to note that unpublished reports should be considered as preliminary data, as the analysis may change in the final publications. The comprehensive summaries have been generated from the oral presentation and the webcasts of the European Society of Cardiology and should provide the readers with the most comprehensive information of relevant publications.     

Balance and Boundaries in Grief Counseling: An Intervention Framework for Volunteer Training

Volunteers or paraprofessional counselors are commonly used to provide supportive care to the bereaved. These counselors generally are trained in basic listening skills, providing a generic, nonspecific approach to intervention that remains to be proven effective. The present paper outlines a framework that provides paraprofessionals with a broader model for intervention with the bereaved. Attention to boundaries as a helper and balance in the grief recovery are central to the model. Interventions are described that provide the paraprofessional counselor with more options for tailoring their counseling strategy to the individual. These include techniques that are presumed to be more specific to the enhancement of grief recovery.     

A new method for the simultaneous analysis of unmodified and modified urinary nucleosides and nucleobases by high performance liquid chromatography

Details are given of a new, rapid and simple pre-fractionation method and an isocratic high performance liquid chromatography system suitable for parallel analysis of nucleosides and nucleobases from urine and other biological fluids. The quantitative recovery and excellent reproducibility of the method is demonstrated by analysis of representative standard RNA catabolites. The advantage of this new method for application to biological samples is discussed.     

Adsorption of Charged Non-albumin Bound Drugs by Amorphous Silica

We investigated the in vitro drug adsorption of PQ 10150 sodium silicate gel (AIS, Santa Clara, CA) with particle size of 230 um and surface area of 400 nr/g. We observed 99% to 88% adsorption of gentamicin; a mean 91 % of disopyramide; a mean 89% of quinidine at low concentration, falling to 75% at higher concentration. Insulin was 88% adsorbed at low concentrations but less so (65%) at higher concentrations. We observed a mean 83 % adsorption of procainamide, a mean 84% of N-acetyl procainamide, 74% oflidocaine, 73% of amitriptyline; and 44% of desipramine. We found an average 14% reduction of total digoxin concentration when serum containing digoxin (2 to 33 ng/mL) was exposed to sodium silicate, while the reduction in free digoxin concentration was 16%. Five percent ethosuximide was also removed. The adsorption of theophylline, phenobarbital, acetaminophen, phenytoin, ethylene glycol, methotrexate, salicylate, thiocyanate and diazepam was minimal and not significant. We conclude that significant amounts of charged, non-albumin bound drugs can be removed by PQ 10150 sodium silicate gel.