阿魏酸钠对抗Aβ_（25-35）诱导的鼠神经元凋亡作用研究

目的探讨阿魏酸钠对β淀粉样蛋白（Aβ25-35）通过谷氨酸诱导的鼠皮层神经元凋亡的影响。方法 培养的皮层神经元分别与谷氨酸（20μmol/L）、Aβ25-35（5μmol/L）、Aβ25-35（5μmol/L）＋谷氨酸（20μmol/L）孵育,采用Hoechst 33258荧光染色法分析神经细胞凋亡;然后用谷氨酸（50μmol/L）诱导神经元凋亡,采用荧光染色法和Western blot观察阿魏酸钠的保护作用。结果单独应用Aβ25-35（5μmol/L）和单独加入谷氨酸（20μmol/L）引起的皮层神经元凋亡率与对照组无明显差异;Aβ25-35与皮层神经元共同孵育5天,接着用谷氨酸处理24h,细胞凋亡率从正常的9.2%±1.5%增加到43%±8%;阿魏酸钠能够显著降低谷氨酸诱导的神经细胞凋亡百分比至21%±5%,并对抗谷氨酸引起的Bcl-2蛋白表达的降低。结论阿魏酸钠能够减弱Aβ25-35提高谷氨酸毒性诱导的鼠皮层神经元凋亡。     

甲基苯丙胺对体外培养的海马神经细胞的毒性作用

目的：通过体外海马细胞培养模型研究MA的作用机制，为MA的研究提供一条新的思路。方法：用浓度分别为0、0．8、1．6、3．2、6．4、12．8mmoL／L的MA处理海马神经元48h，用MTT比色法分析细胞存活率，倒置显微镜和电镜观察凋亡的形态学改变，流式细胞仪检测细胞凋亡百分率。结果：MA可剂量依赖性的降低神经元的存活率；倒置显微镜观察可见神经元胞体固缩，突起断裂，网络消失；电镜可见其细胞核明显固缩和凝聚；流式细胞仪检查可见在0．8、1．6、3．2、6．4、12．8mmol／L MA处理海马神经元，细胞的凋亡率与对照组相比均有显著性差异（P〈0．01），且随剂量增加其凋亡百分率逐渐增大（P〈0．01）。结论：一定浓度的MA对体外培养的海马神经细胞具有细胞毒性，并且诱导神经细胞死亡具有剂量相关效应。     

体外定向化学诱导成人脂肪源性间充质干细胞分化为心肌样细胞

目的：研究在体外不同浓度、不同作用时间诱导条件下5-氮胞苷（5-aza）对脂肪间充质干细胞（ADMSCs）诱导分化为心肌细胞的影响。方法：酶消化筛选法体外分离ADMSCs并传代培养，流式细胞仪鉴定细胞CD44、CD34的表达，不同浓度5-aza（1、5、10、15、20μmol／L）分别诱导12、24、48、72h，相差显微镜下观察细胞形态变化；诱导后4周时免疫细胞化学鉴定心肌特异性肌钙蛋白Ⅰ（cTn—Ⅰ）的表达，分析细胞转化率。结果：体外分离培养的ADMSCs表达CD44，不表达CD34；5-aza诱导后4周时免疫细胞化学显示cTn—Ⅰ表达阳性，以10μmol／L5-aza诱导24h为适宜诱导条件；结论：体外5-aza化学诱导下成人脂肪组织间充质干细胞具有向心肌细胞分化的潜能。     

神经酰胺上调bax表达诱导人内皮细胞凋亡

目的观察外源性神经酰胺（C2-ceramide）对内皮细胞凋亡及相关基因bax表达变化的影响。方法原代培养人脐静脉血管内皮细胞,用不同浓度的ceramide干预内皮细胞24h,并用同一浓度的ceramide作用内皮细胞不同时间后,采用TUNEL法检测细胞凋亡,逆转录聚合酶链反应（RT—PCR）、Westernblot技术对baxmRNA及蛋白表达分析。结果神经酰胺处理组内皮细胞的凋亡阳性率显著高于对照组（P〈0．05）,且具有时间和剂量依赖性;对照组和5、12．5、25、50μmol／L神经酰胺处理24h后,各组细胞凋亡率随干预剂量的加大逐渐上升;终浓度25μmol／L的神经酰胺处理6、24、48h后,细胞凋亡率随时间延长而逐渐升高;神经酰胺处理组baxmRNA及蛋白的表达较对照组显著升高（P〈0．01～0．05）。结论外源性神经酰胺能通过上调bax表达诱导内皮细胞凋亡。     

异丙酚对N-甲基-D-天冬氨酸致PC12细胞损伤时氨基酸水平的影响

目的 观察静脉麻醉药异丙酚对N-甲基-D-天冬氨酸（NMDA）致PC12细胞损伤时氨基酸水平的影响，并探讨异丙酚细胞保护作用的可能机制。方法 建立300μmol／L NMDA诱导的PC12细胞损伤模型，采用高效液相色谱分析法测定PC12细胞氨基酸含量。结果300μmol／L NMDA处理4h可使PCI2细胞谷氨酸含量明显增加，但对天冬氨酸、谷氨酰胺、甘氨酸含量无明显影响；12．5和125μmol／L异丙酚与300μmol／L NMDA同时处理PCI2细胞4h后，谷氨酸含量较单纯NMDA组明显降低（P〈0．05）。结论 异丙酚可明显抑制NMDA诱导PC12细胞损伤所致的谷氨酸合成或释放，提示异丙酚可能通过抑制谷氨酸的合成或释放而产生细胞保护作用。     

组织蛋白酶B在β-淀粉样蛋白刺激星形胶质细胞炎性反应中的作用

目的：研究组织蛋白酶B（Cathepsin B,CB）与β-淀粉样蛋白（β-amyloid protein,Aβ）激活星形胶质细胞诱导炎性反应的关系。方法：应用免疫细胞化学方法鉴定体外培养的星形胶质细胞;四唑盐法检测细胞活性,加入不同浓度溶解状态的Aβ1-40后观察细胞形态学改变,western blot法测定细胞上清CB水平。结果：在5~60μmol/L溶解状态的Aβ1-40作用24 h的条件下,星形胶质细胞的活性没有受到影响,其细胞形态无明显变化,但是随着时间的延长星形胶质细胞逐渐出现肿胀、坏死。40~60μmol/L的溶解状态Aβ1-40诱导的星形胶质细胞上清中检测到CB。结论：Aβ1-40可激活星形胶质细胞,诱导其释放CB,提示CB的神经毒性作用可能是通过诱导神经细胞凋亡实现的。     

基于高内涵分析技术的新型PPARα/γ双重激动剂C333H和P633H的肝细胞毒性初步研究

目的基于建立的高内涵细胞多参数毒性分析方法,观察新型PPARα/γ双重激动剂C333H和P633H的HepG2肝细胞毒性,并初步探查其可能的损伤机制。方法在HepG2人肝癌细胞上,采用高内涵分析（HCA）技术,进行C333H和P633H的细胞毒性（细胞数量、核固缩、膜通透性、线粒体膜电位和细胞色素C）,以及氧化应激和DNA损伤的多参数分析。结果 MTT法测得C333H和P633H抑制HepG2细胞增殖的IC50分别为（161.57±15.29）μmol/L和（101.08±17.58）μmol/L。HCA研究表明C333H在10～100μmol/L,P633H在3～30μmol/L显著降低膜通透性和线粒体膜电位;C333H在30～300μmol/L、P633H在10～100μmol/L浓度依赖性地降低细胞数量。至高浓度时,300μmol/L C333H和100μmol/L P633H进一步显著降低线粒体膜电位和细胞数量,显著增加核DNA及细胞色素C含量,显著减少核面积,此浓度下膜通透性和超氧化物歧化酶显著增加。结论 C333H在低于100μmol/L,P633H在低于30μmol/L时对HepG2肝细胞无明显的毒性反应;线粒体损伤、膜完整性破坏是两化合物引起细胞毒性反应的主要机制,其中线粒体损伤为毒性反应早期敏感性检测指标。此外,HCA法与MTT法在细胞增殖作用的分析上具有较好的一致性。     

奥氮平对淀粉样β蛋白25-35诱导的PC12细胞凋亡的保护作用

目的探讨奥氮平对淀粉样β蛋白25-35（Aβ25-35）诱导的PC12细胞凋亡的保护作用机制。方法以Aβ25-35诱导PC12细胞凋亡,采用MTT比色分析测定细胞存活率,应用Western blot检测Aβ25-35以及奥氮平对PC12细胞Bax、Caspase-3表达的影响,结果10^-14～10^-5mol／L的Aβ25-35减低PC12细胞存活率,50μmol／L及100μmol／I。奥氮平预处理24h可提高PC12细胞存活率,相同浓度Aβ25-35奥氮平预处理组与非处理组比较差异显著（P〈0．05）;0．01、2、20μmol／L Aβ25-35处理的PC12细胞Bax表达增加,50μmol／L奥氮平预处理使0．01μmol／L和20μmol／L Aβ25-35诱导的PC12细胞Bax的表达减低（P〈O．05）;2μmol／L及20μmol／L Aβ25-35处理的PC12细胞Caspase-3的表达增高,5μmol／L奥氮平预处理能抑制其表达升高,与非预处理比较差异显著（P〈O．05）。结论奥氮平可对抗Aβ25-35对PC12细胞的毒性作用,其机制可能与抑制Bax、Caspase-3的表达有关。     

急性心肌梗死患者血清HCY水平动态变化研究

目的：探讨急性心肌梗死（AMI）患者血清同型半胱氨酸（HCY）水平动态变化的临床意义。方法：选择AMI 34例,分别在发病后6 h、24 h、48 h、72 h、7天和21天,动态监测其血清HCY水平变化。结果：AMI患者血清HCY发病后6 h为（22.91±15.56）μmol/L,24 h达峰值（25.98±14.21）μmol/L,48 h为（25.07±13.43）μmol/L,72 h为（23.18±14.54）μmol/L,7天为（21.8±12.16）μmol/L,21天降至（11.98±4.98）μmol/L;ST段抬高患者HCY为（52.17±7.50）μmol/L,非ST段抬高梗死患者为（16.61±6.91）μmol/L,两者比较差异非常显著（P〈0.01）。结论：血清HCY可用于判断心肌梗死急性期心肌损伤程度及预后。     

阿魏酸钠对ox-LDL诱导血管内皮细胞凋亡的影响

目的观察阿魏酸钠(SF)对氧化修饰的低密度脂蛋白(ox-LDL)诱导血管内皮细胞凋亡的保护作用。方法将体外培养的人脐静脉内皮细胞(HUVEC)分为5组,分别为对照组,ox-LDL(50mg/L)组,SF低浓度(5μmol/L)组,SF中浓度(10μmol/L)组及SF高浓度(20μmol/L)组。SF低、中、高浓度组在加入含相应浓度的SF培养1h后,加入终浓度为50mg/L的ox-LDL。采用MTT比色法确定SF的作用浓度。分别采用Hochest 33258细胞核荧光染色法(定性)和流式细胞仪(定量)检测SF对ox-LDL致内皮细胞凋亡的影响。结果一定浓度的SF能够抑制ox-LDL引起的HUVEC存活率降低,对HUVEC损伤具有拮抗作用,40μmol/L以下是SF拮抗细胞损伤的最佳浓度范围。Hochest33258染色后ox-LDL组细胞核显示较强蓝色荧光,可见核固缩、凋亡小体等典型的凋亡形态学特征;加入20μmol/L SF后,细胞核基本恢复其正常形态。流式细胞术结果显示,加入低、中、高浓度的SF后,其细胞凋亡率分别为25.21%±4.07%、12.45%±2.65%、10.85%±1.23%,与ox-LDL组(31.14%±4.29%)比较均明显降低(P<0.05),且存在量效关系。结论 SF具有拮抗ox-LDL诱导的HUVEC损伤,减少血管内皮细胞凋亡的作用。     

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.