ERK1/2信号通路在小鼠库普弗细胞-内毒素反应中的活化规律及其作用

目的：研究内毒素诱导下小鼠库普弗细胞（KC）ERK1／2活化规律、肿瘤坏死因子α（TNFα）分泌规律以及ERK1／2信号通路在TNFα分泌中的作用，探讨防治内毒素血症的新方法。方法：磷酸化ERK时效组以含有100ng/mlLPS 的培养基分别培养KC0，5，10，20，30，45，60，120min；磷酸化ERK量效组分别用含有0，0．1，1，10，100，1000，10000ng/mlLPS的培养基孵育KC30min，免疫印迹杂交检测内毒素诱导KC ERPK1／2活化规律，酶联免疫分析检测内毒素诱导KC释放TNFα规律。以0，0．5，1，10，25，50μmol/L PD98059（ERK信号通路特异性阻断剂）预处理KC1h，酶联免疫分析检测PD98059对LPS诱导TNFα分泌的抑制作用。结果：100ng/ml内毒素刺激后，KC同ERK1／2被迅速活化，30min达到高峰，2h后基本恢复至正常水平；在10pg/ml-100ng/ml范围内，内毒素对ERK1／2激酶具有剂量依赖性的激活作用；内毒素诱导KC TNFα释放显著增加，呈显著的剂量依赖关系；随着PD98059剂量的增加，其对LPS诱导KC TNFα分泌的抑制作用较大。结论：内毒素可诱导KC ERK1／2活化和TNFα分泌，ERK1／2对内毒素诱导KC分泌TNFα具有显著的调节作用，ERK1／2可能是治疗内毒素血症的新靶位。     

内毒素及其增敏系统在烫伤后金葡菌脓毒症中的改变及意义

利用大鼠20％体表面积Ⅲ度烫伤复合金葡菌攻击致脓毒症模型，探讨内毒素及其增敏系统－脂多糖结合蛋白（LBP）和脂多糖受体CD14在G＋菌脓毒症中的变化规律及其与机体过度炎症反应的关系。70只Wistar大鼠随机分为正常对照组（10只），烫伤对照组（10只）和烫伤脓毒症组（50只），检测动物心，肝，肺，肾等组织中LBP，CD14和TNF－α mRNA表达的改变，同时测定组织和血浆内毒素水平，小肠组织二氨氧化酶（DAO）的活性及病理形态学改变，结果显示，烫伤合并金葡菌感染后动物小肠组织DAO活性显著降低，并且至感染后24h仍处于较低水平，同时血浆内毒素水平明显升高，于金葡菌攻击后2h达峰值（P＜0．05），6h则迅速恢复至正常范围，心，肝，肺，肾等组织中LBP，CD14和TNF－α，mRNA表达广泛上调，至24h仍维持于较高水平，肝脏LBP mRNA与肝组织TNF－α基因表达的改变呈显著正相关，提示烫伤后金葡菌感染可导致肠源性内毒素移位和内毒素增敏系统LBP，CD14表达广泛上调，后者作为金葡菌致病因子和内毒素的共同受体对金葡菌脓毒症的发生，发展可能具有促进作用。     

杀菌性/通透性增加蛋白模拟肽对内毒素致血管内皮细胞损伤的保护作用

目的：通过杀菌性／通透性增加蛋白（BPI）模拟肽（BNEP）拮抗内毒素（LPS），观察对血管内皮细胞分泌白细胞介素－6（IL－6），肿瘤坏死因子－α（INF－α）以及表达细胞间黏附分子－1（ICAM－1）的影响，探讨BNEP对LPS致血管内皮细胞损伤的保护作用。方法：采用体外培养的原代人脐静脉内皮细胞（HUVEC）为模型，分别观察单纯应用LPS，BNEP以及不同剂量BNEP与LPS作用后各细胞因子的分泌表达，IL－6，TNF－α应用双抗夹心酶联免疫吸附法（ELISA）测定。ICAM－1采用生物素－抗生物素－过氧化物酶复合物（SABC）－cy3免疫荧光染色，激光共聚焦扫描显微镜下观察其表达情况，结果：LPS 10ng/ml可使IL－6分泌量显著增加；BNEP 60ug/ml即可显著降低10ng/ml LPS诱导的IL－6分泌，随BNEP浓度增加IL－6分泌量呈降低趋势，达240ug/ml时，IL－6分泌量与单纯应用BNEP组比较，差异无显著性意义（P＞0．05），BNEP 120ug/ml与LPS10ng/ml作用后，可使TNF－α分泌量降低94％，BNEP 240ug/ml可完全阻断TNF－α的分泌。激光共聚焦扫描显微镜观察结果显示，BNEP与LPS作用后，ICAM－1在细胞膜及细胞浆的表达明显减弱。结论：BNEP在阻断LPS对细胞的激活和损伤，减少炎症介质和细胞因子的过度释放中具有明显的保护作用。     

库普弗细胞清道夫受体、CD14表达变化及其与内毒素性肝损伤的关系

目的　从细胞受体水平探讨内毒素性肝损伤过程中肝内库普弗细胞 (KC)由防御性逐步转化为效应性的机制。　方法　经尾静脉注射不同剂量内毒素 (LPS)复制轻、中、重度内毒素血症小鼠模型。肝内KC表面清道夫受体 (SR)、CD14表达测定采用免疫组化法 ,并进行计算机图像分析。　结果　注射LPS后 1h ,高剂量组肝内KC表面SR表达明显减少 ,至 3h ,低剂量和中剂量组KC表面SR表达也开始明显减少 ,并随观察时间延长 ,3组KC表面SR表达均进行性减少 ,3组间SR的吸光度差异有显著性意义。 3组肝内KC表面CD14表达在注射LPS后 1h均明显增多 ,并随时间延长而更加明显 ,但 3组间CD14的吸光度无统计学意义。相关分析显示 ,各剂量组肝内SR与CD14表达变化呈显著负相关。血浆丙氨酸氨基转移酶 (ALT)、总胆红素 (TB)和肝组织内丙二醛 (MDA)水平变化分别与SR表达下调呈显著负相关 ,与CD14表达上调呈显著正相关。肝组织结构改变及动物死亡率也与肝内KC表面SR、CD14表达变化呈明显的平行关系。　结论　内毒素致肝损伤过程中 ,肝内KC表面SR表达下调、CD14表达上调可能是KC防御功能减弱、致炎作用增强的重要机制。     

固定化亲和吸附剂血液灌流对脓毒症大鼠作用的研究

观察固定化多粘菌素B血液灌流对脓毒症大鼠血液中内毒素，TNF－α和血液流变学的作用。将24只Wistar大鼠分为固定化多粘菌素B组（LPS＋HP＋PMB组）、空灌流组（LPS＋HP组）和对照组（LPS模型组），给大鼠一次性静注内毒素（LPS）0．5mg/kg，LPS＋HP＋PMB组、LPS＋HP组注入内毒素4h后抽出血液，分离出血浆，将血浆经特异亲和剂灌流柱或未偶联亲和剂的空灌流柱进行血液灌流，灌流后浆与细胞混匀回输动物。测定血浆中LPS、TNF－α水平及全血粘度和红细胞比容变化。结果发现，经特异性亲和灌流柱灌流1h，血浆LPS、TNF－α水平明显下降，提示固定化多粘菌素B血液灌流能显著清除血浆中内毒素，改善血液流变学。     

AP—2在内毒素刺激大鼠肺泡巨噬细胞中的动态变化及其在TNF—α表达中的调控作用

为观察内毒素刺激对大鼠肺泡巨噬细胞（AM）中转录因子AP－1活性的影响，探讨AP－1在LPS刺激大鼠肺泡巨噬细胞表达TNF－α的基因调控作用，应用凝胶迁移阻滞技术（EMSA）检测LPS（E coli 026:B6)刺激大鼠AM核蛋白中AP－1的DNA结合活性的动态变化，在LPS刺激前12h，应用转基因技术将AP－1的硫代寡核苷酸圈套（S－ODN decoy)转入大鼠AM，应用ELISA法观察AP－1硫代寡核苷酸圈套对LPS刺激大鼠AM表达TNF－α的影响，结果发现，应用100ng/ml LPS刺激大鼠AM 0．5h，AP－1即迅速出现活化并达到峰值，在1h活性略有下降，3h活性又逐渐回升，5h,8h又迅速回落，AP－1的活化状态至少可以持续8h，且与LPS刺激呈明显的量效关系；AP－1的S－ODN圈套能明显抑制但不能完全阻断LPS诱导的TNF－α表达，说明LPS刺激可使大鼠AM内AP－1活化，其在LPS介导的炎症反应中可能发挥重要作用，AP－1在LPS诱导的大鼠AM表达TNF－α中可能起重要的调控作用，但TNF－α表达可能还与其他核转录因子有关。     

CD14和SR-A蛋白表达与内毒素介导的急性肝损伤的关系

目的 观察肝组织清道夫受体A(SR-A)和脂多糖(LPS)受体CD14在内毒素介导的急性肝损伤中的表达及其意义.方法 Wistar大鼠90只,随机均分为内毒素血症(LPS)组和生理盐水注射(NS)组,LPS组经尾静脉注射LPS 5rng/kg,建立内毒素血症和急性肝损伤动物模型.NS组尾静脉注射0.2ml生理盐水作为对照.分别于注射后0、1.5、3、6、12h检测血浆LPS、TNF-α、IL-1、丙氨酸转氨酶(ALT)及总胆红素(TB)含量,肝组织中SR-A和CD14蛋白表达情况.光镜观察肝组织病理变化,电镜观察库普弗细胞(KCs)超微结构的变化,并测定KCs的吞噬功能.结果 随时间延长,LPS组KCs从形态和功能上呈现激活改变(数量增多、体积增大、吞噬功能增强),血浆TNF-α、IL-1含量明显增加,肝细胞变性、坏死等病理改变以及肝功能损害进行性加重,同时肝组织中SR-A表达明显下降、CD14表达明显升高(P<0.01).结论 肝组织中进行性的SR-A表达下降和CD14表达增强可能是内毒素介导急性肝损伤的重要机制之一.     

内毒素作用下淋巴细胞中CD4＋CD25＋调节性T细胞变化及连翘对其的影响

目的探讨不同时间点内毒素（LPS）对CD4＋CD25＋调节性T细胞（CD4＋CD25＋regulatory T cell,Treg）的影响及连翘（forsythia suspensa,FS）干预后CD4＋CD25＋调节性T细胞的变化。方法体外培养Wistar大鼠脾脏淋巴细胞,分为对照组、LPS（10ng/ml）组、LPS（10ng/ml）＋多黏菌素B组（10ng/ml）、LPS（10ng/ml）＋连翘（12．5、25、50mg／ml）组,分别于3、6、12h收集标本．流式细胞术检测CD4＋CD25＋调节性T细胞在淋巴细胞中的百分比,荧光定量RT—PCR法检测Foxp3mRNA表达,酶联免疫吸附试验（ELISA）法检测TNF—a、IL-10分泌水平。结果脾脏淋巴细胞中CD4＋CD25＋Treg的百分比和Foxp3mRNA的表达在12h较3、6h明显升高（P〈0．01）;TNF—a、IL-10的分泌随时间点的延长均显著升高（P〈0．01）;随连翘剂量增加,TNF-a、IL-10分泌水平,Foxp3mRNA表达和CD4＋CD25＋Treg百分比均明显降低（P〈0．05）。结论内毒素参与并诱导了CD4＋CD25＋Treg在淋巴细胞中的变化,其随内毒素作用时间延长呈上升趋势。连翘能显著降低CD4＋CD25＋Treg在淋巴细胞中的百分比。     

已酮可可碱对脾切除大鼠内毒素血症的干预作用

目的与方法：通过动态观察已酮可可碱干预处理脾切除后大鼠内毒素血症时肝及肺组织TNA－α水平及肝、肺组织TNF－αmRNA表达的变化,探讨PTX对无脾动物脓毒症的保护作用及机制。结果：内毒素攻击后无脾动物肝肺组织TNF－α水平、TNF－αmRNA的表达及肺组织髓过氧化物酶水平均较有脾动物显著升高（P＜0．05）,PTX预处理可显著抑制这种增高（P＜0．05）。结论：PTX通过下调肝肺组织TNF－αmRNA的表达,减少组织中TNF－α浓度对脾切除后凶险性感染（OPSI）的防治及脓毒症的治疗可能有益。     

单磷酸类脂A对内毒素血症小鼠的保护及对丙二醛的作用

目的 探讨单磷酸类脂A（MPLA）对内毒素血症小鼠的预防性保护作用及其与丙二醛的关系。方法 观察MPLA预处理对内毒素血症小鼠存活率的影响以及MPLA预处理对内毒素血症小鼠血、肝和肺组织丙二醛（MDA）含量变化的影响，并且行肝和肺组织病理学检查。结果 MPLA（100μg）3天前预处理能明显提高内毒素血症小鼠24小时存活率（60％：25％，P＜0．05）。内毒素血症小鼠血、肝及肺组织中MDA水平早期就显著升高，而MPLA预处理则可显著抑制其升高并可减轻肝及肝组织的组织病理改变。结论 MPLA预处理可提高内毒素血症小鼠的存活率，并且能抑制MDA的水平以及减轻组织病理改变。     

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.     

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).     

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.     

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.     

Bronchial stump fistula :treatment with covered retrievable hinged metallic stents-preliminary clinical experience

Objective To evaluate the preliminaily clinical efficacy and retrievability of a retrievable hinged covered metallic stent in the treatment of the bronchial stump fistula （BSF）. Methods Between April 2003 and March 2005, 8 patients with bronchial stump fistula after pneumonectomy or lobectomy were treated with two types （A and B） of retrievable hinged covered metallic stents. Type A stent was placed in 6 patients and type B in 2 under fluoroscopic guidance. The stent was removed with a retrieval set when BSF was healed or complications occurred. Results Stent placement in the bronchial tree was technically successful in all patients, without procedure-related complications. Immediate closure of the BSF was achieved in all patients after the procedure. Stents were removed from all patients but one. Removal of the stents was difficult in two patients due to tissue hyperplasia. Patients were followed up for 6 - 21 months. Placement of the stents remained stable in all patients except one due to severe cough. Permanent closure of BSF was achieved in 7 （87.5%） of 8 patients. Conclusion Use of a retrievable hinged covered expandable metallic stent is a simple, safe, and effective procedure for closure of the BSF. Retrieval of the stent seems to be feasible. （J Intervent Radiol, 2007, 16： 253-257）     

Prevalence and severity of hip and groin pain in sub‐elite male football: a cross‐sectional cohort study of 695 players

The purpose of this study was twofold: (a) to investigate the prevalence of hip and groin pain in sub‐elite male adult football in Denmark and (b) to explore the association between prevalence and duration of hip and groin pain in the previous season with the Copenhagen Hip and Groin Outcome Score (HAGOS) in the beginning of the new season. In total 695 respondents from 40 teams (Division 1–4) were included. Players completed in the beginning of the new season (July–Sept 2011) a self‐reported paper questionnaire on hip and/or groin pain during the previous season and HAGOS. In total 49% (95% CI: 45–52%) reported hip and/or groin pain during the previous season. Of these, 31% (95% CI: 26–36%) reported pain for >6 weeks. Players with the longest duration of pain during the previous season had the lowest HAGOS scores, when assessed at the beginning of the new season, P < 0.001. This study documents that half of sub‐elite male adult football players report pain in the hip and/or groin during a football season. The football players with the longest duration of pain in previous season displayed the lowest HAGOS scores in the beginning of the new season.