褐藻糖胶对照射大鼠免疫功能和淋巴细胞凋亡影响

目的　研究不同剂量褐藻糖胶对受γ射线照射大鼠免疫功能影响的作用机理。方法不同剂量褐藻糖胶经口给予实验组大鼠 ,连续给药 10d后行一次性 6 0Coγ射线照射 ,18h后测定各组大鼠体液免疫、细胞免疫和非特异性免疫相关指标 ,并测定大鼠脾淋巴细胞凋亡率。结果　 10 0mg kg褐藻糖胶即能显著提高照射后大鼠T、B淋巴细胞增殖能力、巨噬细胞吞噬功能、血清溶血素含量 ,增强大鼠迟发性超敏反应 ,抑制照射后脾淋巴细胞凋亡 ,并呈现剂量 效应关系 ,与阳性对照组比较差异有非常显著性 (P <0 0 1)。结论　褐藻糖胶是一种抗照射免疫调节剂 ,对照射后大鼠的免疫系统具有恢复作用 ,其抗照射作用与抑制淋巴细胞凋亡有关     

放射复合伤口愈合中外周血淋巴细胞凋亡及其与愈合延迟的关系

为研究放射复合伤口愈合中外周血淋巴细胞凋亡规律并探讨其与愈合延迟的关系，将120只大鼠随机分为单纯创伤组与创伤复合照射组，用原位末端标记（TUNEL）方法检测细胞凋亡，碱磷酶免疫组织化学方法检测Bax,Bcl-2蛋白表达，结果发现，创伤复合照射组动物白细胞数出现下降，伤后3天降至最低，伤后5天仍显著低于单纯创伤组；伤后两组动物中血淋巴细胞凋亡率均升高，但创伤照射组凋亡率始终高于单纯创伤组；伤后不同时间淋巴细胞Bax蛋白出现升高，创伤照射组明显高于单纯创伤组，而Bcl-2呈现相反的变化趋势，该结果提示，照射后外周血白细胞数的降低和淋巴细胞凋亡的增加是辐射延迟伤口愈合的重要原因，Bax和Bcl-2蛋白参与了淋巴细胞凋亡的调控。     

重离子辐射对人外周血T淋巴细胞生物学性能的影响

目的研究重离子对人外周血T淋巴细胞增殖、凋亡等生物学性能的影响并探讨其机制,为肿瘤放射治疗的辐射防护提供实验依据和基础。方法 Ficoll分离法分离人外周血T淋巴细胞,采用12C重离子束坪区照射,照射样品能量为70 MeV、LET=29 keV/μm,照射剂量为1.0和2.0 Gy,剂量率为0.5 Gy/min。分别于照射后12、24h,RT-PCR检测凋亡相关基因Bcl-2,Bax,Caspase3,Caspase8和Caspase9的表达;于照射后24、48 h,CCK8法检测细胞增殖能力;于照射后24、48 h,采用AnnexinV-PE/7-AAD、AnnexinV-FITC/PI法检测凋亡发生,并采用RT-PCR检测凋亡相关蛋白Bcl-2、Bax和Caspase3的表达。结果重离子照射可明显抑制人外周血T淋巴细胞的增殖,随着剂量增大,抑制作用更加明显。同时,重离子照射可促进T淋巴细胞的凋亡,特别是对于晚期凋亡的诱导作用（P〈0.01）。RT-PCR检测结果显示,重离子辐射可抑制抗凋亡蛋白Bcl-2的表达,促进促凋亡蛋白Bax和Caspase3的表达（P〈0.01）。结论重离子辐射可显著影响抑制T淋巴细胞的增殖并促进其凋亡。     

补充Ebselen对运动训练大鼠心肌Bcl-2和Bax蛋白表达、部分自由基代谢及超微结构的影响

目的:探讨补充ebselen对运动训练大鼠心肌细胞凋亡、自由基代谢及超微结构的影响。方法:健康雄性SD大鼠30只,适应性饲养、筛选,淘汰个别不适应跑台训练者,将剩余大鼠随机分为3组:安静对照组8只、运动对照组8只、运动+给药组8只;后两组进行为期8周的跑台训练。运动+给药组大鼠灌服溶于5%二甲基亚砜生理盐水的ebselen溶液,用量为50mg/kg/d,其它两组灌服同等量的DMSO生理盐水作为对照。测定心肌组织Bcl-2、Bax蛋白表达灰度值和超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性和丙二醛(MDA)的含量,透射电子显微镜观察心肌超微结构。结果:与安静对照组相比,运动对照组SOD、GSH-Px表达显著下降(P<0.05,P<0.01),MDA含量显著升高(P<0.01),Bax/Bcl-2蛋白表达比值显著升高(灰度值比值显著降低)(P<0.01),透射电镜观察显示心肌结构损伤明显;运动+给药组大鼠SOD、GSH-Px表达显著升高(P<0.01),MDA含量显著降低(P<0.01),Bax/Bcl-2蛋白表达比值显著降低(灰度值比值显著升高),透射电镜显示大鼠心肌细胞结构基本正常。结论:Ebselen对运动训练导致的大鼠心肌损伤有明显保护作用,其机制可能与ebselen可以抑制运动诱导的Bax蛋白过度表达、Bcl-2蛋白表达的减少以及提高机体抗氧化酶活性有关。     

NF-kB抑制与X射线诱导的人非霍奇金淋巴瘤细胞凋亡的研究

目的 探讨NF-kB p65对X射线诱导人非霍奇金淋巴瘤(non-Hodgkin lymphoma,NHL)细胞凋亡的作用及其调控机制.方法 以NF-kB p65抑制剂quinazoline (QNZ)处理人NHL细胞.将NHL细胞株Namalwa、Ramos和Raji细胞分为空白对照组、单纯照射组(IR)和X射线+QNZ实验组( IR+ QNZ),采用Annexin-V染色方法检测肿瘤细胞凋亡水平的变化,采用Western blot方法检测各细胞株中Survivin及凋亡相关蛋白Bax、Bcl-2和Cleaved Caspase-3的表达水平;应用实时定量PCR方法检测Survivin mRNA水平.结果 应用流式细胞仪检测凋亡细胞百分比,结果显示,QNZ处理后进行照射与单纯照射组相比凋亡细胞明显增加,且具有药物浓度依赖性(t=2.93～12.52,P＜0.05),Western blot法检测结果显示,电离辐射可显著增加人NHL细胞中Survivin蛋白的表达.而应用1、10和50 nmol/L QNZ处理人NHL细胞24 h后进行照射,可显著下调电离辐射所诱导的NHL细胞中的Survivin蛋白表达.随药物浓度增加其作用更为明显(t=3.29～ 16.72,P＜0.05).同时凋亡相关蛋白Bcl-2表达减低,而Bax和Cleaved caspase-3蛋白表达增加,Bcl-2/Bax比值明显降低,与单纯照射组相比,差异有统计学意义(t =6.20 ～9.91,P＜0.05).预处理QNZ后射线诱导的3种NHL细胞Survivin mRNA均不同程度下降.结论 抑制NF-kB能够增加X射线诱导的NHL细胞凋亡,其机制可能与下调Survivin蛋白表达水平及对凋亡相关蛋白Bcl-2家族的调节有关.     

c-ski对大鼠皮肤成纤维细胞抗辐射作用的影响

目的 研究原癌基因c—ski对大鼠皮肤成纤维细胞抗辐射作用的影响，并探讨其可能的机理。方法 采用Annexin—V—FITC—PI标记的方法。通过流式细胞仪检测2—8Gy的软X线照射对培养的大鼠皮肤成纤维细胞凋亡的影响；观察转染c—ski基因后细胞凋亡的变化；并采用Western blot检测c—ski对凋亡蛋白Bax、Bcl-2表达的影响。结果 软X线照射以剂量依赖的方式增加细胞凋亡，并且随着时间的延长进一步增加，大约在36h时达到峰值。转染c—ski基因使4Gy照射24h后的成纤维细胞凋亡率显著下降。转染c—ski基因48h后，Bax的表达和对照组差异无统计学意义，而Bcl-2的表达显著增加。结论 c—ski可降低成纤维细胞的辐射敏感性，促进Bcl-2的表达可能是其机理之一。     

pEgr-hPTEN稳定转染联合辐射诱导人胶质瘤SHG-44细胞凋亡及Bcl-2表达下调

目的 探讨辐射诱导表达载体pEgr-hPTEN体外稳定转染人胶质瘤SHG-44细胞后联合X射线照射，诱导细胞凋亡的作用及凋亡相关蛋白Bcl-2表达的变化。方法 以脂质体介导携有外源野生型PTEN基因的辐射诱导表达载体pEgr-hPTEN，体外转染SHG-44细胞，筛选稳定转染的细胞克隆并扩增培养；应用电子显微镜、流式细胞仪等方法，检测稳定转染联合X射线照射对胶质瘤细胞超微结构、细胞凋亡及Bcl-2蛋白表达等特性的影响。结果 稳定转染细胞超微结构有明显的退行性改变，可见核内染色质趋边的类似早期凋亡的改变；稳定转染联合X射线照射可诱导肿瘤细胞凋亡，5Gy以内随吸收剂量的增加，早期凋亡细胞百分数明显增加，稳定转染不同剂量照射组早期凋亡细胞百分数分别为稳定转染0Gy假照组的1．5—2．3倍、为未转染照射组的1．9—4．4倍、为未转染0Gy假照组的3．4—5．1倍；同时稳定转染细胞Bcl-2蛋白表达则呈剂量依赖性下降。结论 体外PTEN基因转染联合X射线照射可诱导肿瘤细胞凋亡明显增多，Bcl-2蛋白表达下调，具有显著的肿瘤抑制作用。     

Ox-LDL诱导人肾小球系膜细胞凋亡在慢性肾炎中的作用

目的 :探讨氧化低密度脂蛋白 (Ox-LDL)诱导肾小球系膜细胞凋亡的调控机制及其在慢性肾炎中的作用。方法 :通过人肾小球系膜细胞体外培养 ,采用原位末端标记和DNA凝胶电泳检测低浓度的Ox -LDL (0、 2 5、 5 0、10 0 μg/ml)对系膜细胞凋亡的影响 ,免疫组化维生素E干预前后Ox-LDL对系膜细胞Bax/Bcl- 2表达的影响 ;按血浆LDL水平 ,把有不同程度肾小球硬化的慢性肾炎患者肾标本分为LDL正常组和LDL升高组 ,分别采用DNA原位末端标记和免疫组化 ,检测肾小球细胞凋亡及Bax/Bcl- 2蛋白表达。结果 :低浓度Ox-LDL (0～ 10 0 μg/ml)以剂量依赖方式促进肾小球系膜细胞凋亡 ,并使肾小球系膜细胞Bax的蛋白表达呈时间 -剂量依赖性增加 ,P <0 0 1,Bcl- 2的蛋白表达轻度升高后呈时间 -剂量依赖性下降 ,P <0 0 1,导致Bax/Bcl- 2比值增大 ;维生素E干预后这种变化不明显 ,P >0 0 5。LDL升高组肾小球凋亡和Bax蛋白表达明显增加 ,P <0 0 1;Bcl- 2变化不明显 ,P >0 0 5 ,血LDL水平与肾小球硬化指数、凋亡指数、Bax/Bcl- 2比值有显著正相关关系。结论 :Ox -LDL能诱导肾小球系膜细胞凋亡 ;Bax/Bcl- 2比值上升可能是Ox -LDL诱导肾小球系膜细胞凋亡的重要机制 ;脂质异常可能是导致肾小球硬化中细胞减少的重要原因     

去甲斑蝥素诱导人肾癌786—0细胞凋亡的体外研究

目的探讨去甲斑蝥素（NCTD）诱导人肾癌细胞株786—0凋亡的机制。方法实验分NCTD组和对照组．分别应用MTT法、流式细胞术和实时定量PCR检测NCTD对体外培养786—0细胞的生长抑制率、细胞周期和凋亡率以及肿瘤凋亡控制因子Bcl-2、Bax mRNA的表达。结果NCTD能抑制786—0细胞生长,随着NCTD浓度升高、处理时间延长作用增强,有明显的时间-剂量依赖性。流式分析显示,786—0细胞的S期细胞减少,G2/M期细胞增多,凋亡率上升。实时定量PCR结果表明,在80μmol／L NCTD处理24h后,Bcl-2 mRNA表达量明显减少、Bax mRNA表达水平则升高,Bcl-2／Bax比值明显下降,与对照组比较差异有统计学意义（P〈0．05）。结论NCTD可明显抑制786—0细胞的增殖和生长,诱导其凋亡,其机制可能与干扰786—0细胞生长周期、抑制DNA合成代谢以及影响凋亡相关基因Bcl-2和Bax的表达有关。     

葛根素对成年大鼠创伤性脑损伤神经细胞的保护作用

目的 探讨葛根素对成年大鼠创伤性脑损伤神经细胞中Bcl-2、Bax蛋白表达的影响。方法 不同浓度葛根素作用于创伤性脑损伤神经细胞,通过病理学检查、Tunel法及免疫组织化学法检测神经细胞凋亡及Bcl-2、Bax蛋白的表达。结果 葛根素作用脑损伤神经细胞7天后,以浓度效应关系改善创伤性脑损伤神经细胞的病理改变,减轻细胞凋亡程度,增加Bcl-2的表达,降低Bax／Bcl-2比率,但不影响Bax的表达。结论 葛根素对创伤性脑损伤神经细胞具有保护作用,且呈剂量依赖性,其机制可能与葛根素抗细胞凋亡,增加Bcl-2的表达有关。     

Knee injury and Osteoarthritis Outcome Score (KOOS) - validation of a Swedish version

The Knee injury and Osteoarthritis Outcome Score (KOOS) is a self-administered instrument measuring outcome after knee injury at impairment, disability, and handicap level in five subscales. Reliability, validity, and responsiveness of a Swedish version was assessed in 142 patients who underwent arthroscopy because of injury to the menisci, anterior cruciate ligament, or cartilage of the knee. The clinimetric properties were found to be good and comparable to the American version of the KOOS. Comparison to the Short Form-36 and the Lysholm knee scoring scale revealed expected correlations and construct validity. Item by item, symptoms and functional limitations were compared between diagnostic groups. High responsiveness was found three months after arthroscopic partial meniscectomy for all subscales but Activities of Daily Living.     

Endovascular management of carotid-cavernous fistulas

Objective To investigate endovascular treatment of traumatic direct carotid-cavernous fistulas （CCF） and their complications such as pseudoaneurysms. Methods： Over a five-year period, 22 patients with traumatic direct CCFs were treated endovascularly in our institution. Thirteen patients were treated once with the result of CCF occluded, 8 twice and 1 three times. Treatment modalities included balloon occlusion of the CCF, sacrifice of the ipsilateral internal carotid artery with detachable balloon, coll embolization of the cavernous sinus and secondary pseudoaneurysms, and covered-stem management of the pseudoaneurysms. Results All the direct CCFs were successfully managed endovascularly. Four patients developed a pseudoaneurysm after the occlusion of the CCF with an incidence of pseudoaneurysm formation of 18.2% （4/22）. A total number of 8 patients experienced permanent occlusion of the ICA with a rate of ICA occlusion reaching 36.4% （8/22）. Followed up through telephone consultation from 6 months to 5 years, all did well with no recurrence of CCF symptoms and signs. Conclusion Traumatic direct CCFs can be successfully managed with endovascular means. The pseudoaneurysms secondary to the occlusion of the CCFs can be occluded with stent-assisted coiling and implantation of covered stents.     

The acutely limping child

Acute limping may be the result of multiple pathologies in children. The differential diagnosis varies based on the age of the child. Irrespective of age, the initial imaging work-up includes AP and frog leg radiographs of the pelvis and ultrasound; MRI may sometimes be helpful. In children less than 3 years, infections and trauma are most frequent. MRI is the imaging modality of choice when osteomyelitis is clinically suspected. Between the ages of 3 and 10 years, transient synovitis of the hip and Legg-Calvé-Perthes disease are main considerations but infection, inflammation and focal bony lesions are also considered. In children over 10 years, slipped capital femoral epiphysis also is considered.     

Do Balance Board Training Programs Reduce the Risk of Ankle Sprains in Athletes?

Introduction Ankle sprains are the most common musculo-skeletal injury that occurs in athletes,particularly in sports that require jumping and landing on one foot such as soccer,and basketball(1-4).These injuries often result in significant time loss from participation,long-term disability,and have a major impact on health care costs and resources(5-8).     

High Intensity Interval Training:New Insights

KEY POINTS ·High-intensity interval training(HIT)is characterized by repeated sessions of relatively brief，intermittent exercise.often performed with an“a11 out”effort or at an intensity close to that which elicits peak oxygen uptake(i.e.，≥90%of VO2 peak).     

Developmental validation of the PowerPlex(®) ESI 16 and PowerPlex(®) ESI 17 Systems: STR multiplexes for the new European standard

In response to the ENFSI and EDNAP groups’ call for new STR multiplexes for Europe, Promega^® developed a suite of four new DNA profiling kits. This paper describes the developmental validation study performed on the PowerPlex^® ESI 16 (European Standard Investigator 16) and the PowerPlex^® ESI 17 Systems. The PowerPlex^® ESI 16 System combines the 11 loci compatible with the UK National DNA Database^®, contained within the AmpFlSTR^® SGM Plus^® PCR Amplification Kit, with five additional loci: D2S441, D10S1248, D22S1045, D1S1656 and D12S391. The multiplex was designed to reduce the amplicon size of the loci found in the AmpFlSTR^® SGM Plus^® kit. This design facilitates increased robustness and amplification success for the loci used in the national DNA databases created in many countries, when analyzing degraded DNA samples. The PowerPlex^® ESI 17 System amplifies the same loci as the PowerPlex^® ESI 16 System, but with the addition of a primer pair for the SE33 locus. Tests were designed to address the developmental validation guidelines issued by the Scientific Working Group on DNA Analysis Methods (SWGDAM), and those of the DNA Advisory Board (DAB). Samples processed include DNA mixtures, PCR reactions spiked with inhibitors, a sensitivity series, and 306 United Kingdom donor samples to determine concordance with data generated with the AmpFlSTR^® SGM Plus^® kit. Allele frequencies from 242 white Caucasian samples collected in the United Kingdom are also presented. The PowerPlex^® ESI 16 and ESI 17 Systems are robust and sensitive tools, suitable for the analysis of forensic DNA samples. Full profiles were routinely observed with 62.5 pg of a fully heterozygous single source DNA template. This high level of sensitivity was found to impact on mixture analyses, where 54–86% of unique minor contributor alleles were routinely observed in a 1:19 mixture ratio. Improved sensitivity combined with the robustness afforded by smaller amplicons has substantially improved the quantity of data obtained from degraded samples, and the improved chemistry confers exceptional tolerance to high levels of laboratory prepared inhibitors.