LPS诱导肺微血管内皮细胞ICAM-1的表达及核调控机理的实验研究

目的：研究细胞脂多糖（LPS）诱导的肺微血管内皮细胞（PMVEC）ICAM－1的表达及调控机制，方法：100ng.ml^-1LPS刺激PMVEC0h,2h,4h,6h,8h或10ng.ml^-1，50ng.ml^-1,100ng.ml^-1LPS刺激6h，免疫细胞化学检测PMVECICAM－1的表达，凝胶电泳迁移率变化分析检测NF－κB的活化，并通过加入活化阻断剂PDTC观察对PMVEC ICAM－1表达的影响。结果：PMVECICAM－1的表达与LPS的刺激呈时相－剂量依赖方式，LPS的刺激迅速活化NF－κB，60min达到高峰，后逐渐下降，PDTC能显著降低ICAM－1的表达（P＜0．01），结论：LPS刺激诱导NF－κB的活化，启动ICAM－1的合成表达。     

X射线全身照射对小鼠腹腔巨噬细胞CD14和TLR4表达的影响

CD14是LPS/LBP复合物的受体,以LPS-LBP-CD14三联体形式介导LPS诱导细胞活化.TLR4是Toll样受体中发现较早的一个受体,是LPS信号跨入细胞所必需的跨膜受体[1].CD14和TLR4作为巨噬细胞膜上的跨膜受体将细胞外信号传到细胞内,通过一系列蛋白质级联反应激活转录因子NF-κB和Jun/Fos,释放IL-1和IL-6等细胞因子而发生炎性反应.本研究探讨X射线全身照射对小鼠腹腔巨噬细胞CD14和TLR4表达的影响,为临床应用X射线进行抗炎、抗肿瘤治疗提供理论和实验依据.     

核因子—κB在内毒素刺激大鼠肺泡巨噬细胞中的动态变化及其在肿瘤坏死因子—α表达中的作用

目的 观察内毒素(LPS)刺激对大鼠肺泡巨噬细胞(alveolar macrophages，AM)中核因子—κB(NF—κB)活性的影响，探讨NF—κB在LPS刺激大鼠AM表达肿瘤坏死因子—α(刀NF—α)中的作用。方法体外观察LPS刺激大鼠AM中NF—κB活性的动态变化，应用圈套策略观察NF—κB在LPS刺激大鼠AM表达刀NF—α中的作用。结果LPS刺激可使大鼠AM内NF—κB明显活化，并与LPS剂量正相关；抗体超迁移率实验结果显示p50、p65参与了NF—κB的活化；NF—κB的圈套硫代寡核苷酸(S—ODN)能明显抑制LPS刺激大鼠AM表达TNF—α，但不能完全阻断LPS诱导的TNF—α表达。结论 NF—κB(p50／p65)在LPS介导的炎症反应中发挥重要作用，LPS刺激大鼠AM表达TNF—α受NF—κB调控，但其他核转录因子可能也在TNF—α的表达中发挥重要作用。     

核因子-κB圈套寡核苷酸对核因子-κB活性及创伤炎症反应大鼠肝脏功能损害的影响

目的 探讨核因子(NF)-κB圈套寡核苷酸(ODN)对NF—κB活性及创伤炎症反应大鼠肝脏功能损害的影响。方法 利用电泳迁移率改变实验(EMSA)，测定合成的环状哑铃形圈套ODN对NF—κB的DNA结合能力的竞争抑制作用；利用NF—κB反应性报告细胞株HEK-不稳定增强型绿色荧光蛋白(d2EGFP)，观察圈套ODN瞬时转染对报告基因表达的影响。Wistar大鼠96只，随机分为对照组、创伤性炎症组、圈套ODN组和无关ODN组。利用EMSA检测各组肝脏组织NF—κB的DNA结合活性，用逆转录-聚合酶链反应(RT—PCR)检测大鼠肝脏组织TNF—α、白细胞介素-6(IL-6)mRNA水平，酶联免疫吸附实验(ELISA)法检测大鼠肝脏组织肿瘤坏死因子-α(TNF—α)、IL-6的蛋白水平。结果 设计的环状哑铃形圈套ODN对NF—κB的DNA结合能力具有竞争抑制作用，1μg／ml和2μg／ml圈套ODN瞬时转染HEK—d2EGFP，可明显抑制p65蛋白诱导的d2EGFP表达。大鼠创伤性炎症后3h，肝脏NF—κB活性开始升高，伤后12h达高峰。肝脏组织TNF—α、IL-6mRNA水平和蛋白水平也明显上升，与NF—κB的活性改变一致。圈套ODN治疗后，大鼠肝脏组织TNF—α、IL-6的表达明显下降，肝脏功能明显好转，而无关ODN则没有治疗效果。结论 设计的靶向NF—κB的环状哑铃形圈套ODN可通过特异性抑制NF-κB活性，有效阻止创伤性炎症大鼠肝脏炎症介质TNF—α、IL-6的释放，从而改善肝脏功能。     

Toll样受体4及核因子-κB在脂多糖诱导的气道上皮细胞基质金属蛋白酶9表达中的作用

目的 探讨Toll样受体4(TLR4)和核因子(NF)-κB在脂多糖(LPS)诱导的气道上皮细胞基质金属蛋白酶9(MMP-9)表达中的作用,观察TLR4抑制剂TAK-242对MMP-9表达的影响.方法 以终浓度为0、0.1、l、10、100μg/L的LPS刺激HBE4-E6/E7细胞24h,或以LPS(终浓度10μg/...     

人肠上皮细胞内毒素低反应性及其机制探讨

为探讨人正常肠上皮细胞对内毒素（LPS）刺激低反应性的机制，用ELISA法检测LPS刺激肠上皮细胞18h后IL－8的分泌水平，用RT－PCR及RPA法检测肠上皮细胞LPS跨膜信号转导相关受体TLR4，TLR2，CD14和MD2 mRNA的表达及LPS刺激对其表达的影响，结果表明：LPS刺激不能引起肠上皮细胞IL－8的分泌，而IL－1β刺激却能引起其分泌；人正常肠上皮细胞TLR4，TLR2，CD14和MD2mRNA的表达均较弱，LPS刺激后，TLR4，CD145 CD14和MD2 mRNA表达进一步降低，而TLR2的表达无显著变化，说明肠上皮细胞对LPS的低反应性与细胞表达LPS相关受体低表面有一定的内在联系，从而进一步证实TLR4，CD14，MD2在介导LPS跨膜信号转导中起重要作用。     

糖皮质激素抑制辐射诱导P388细胞NF-κB活化

目的 研究糖皮质激素对辐射诱导P388白血病细胞凋亡与NF－κB活化的影响。方法 采用TdT介导的dUTP缺口末端标记技术（TUNEL）检测细胞凋亡；采用电泳迁移率变动分析（EMSA）检测P388细胞NF－κB活化水平。结果 2，4，6和8Gy辐射诱导P388细胞凋亡，同时使NF－κB活化。地塞米松（DXM）增强由2，4，6和8Gy辐射诱导的P388细胞凋亡，抑制由2，4，6和8Gy辐射诱导的P388细胞NF－κB活化，凋亡增加率分别为60％，100％，129％和67％，NF－κB活化抑制率分别为25％，45％，52％和40％。结论 辐射诱导P388白血病细胞凋亡的同时激活NF－κB；糖皮质激素可通过抑制NF－κB活化，增强其诱导白血病细胞凋亡的作用。     

圈套寡核苷酸影响腹腔巨噬细胞炎症介质表达的实验研究

目的 设计合成靶向核因子—κB(NF—κB)的哑铃形圈套寡核苷酸(0DNs)，并检测其抗炎效应。方法 提取大鼠腹腔巨噬细胞(pMφ)，随机分为正常对照组、单纯内毒素(LPS)组、圈套0DNs处理组、无关圈套0DNs处理组和脂质体处理组。收集正常对照组和单纯LPS组LPS刺激后1，2，6，12，18，24k上清及细胞；采用阳离子脂质体按质量比为4：1的比例分别介导2，4，8mg/L圈套0DNs转染大鼠pMφ细胞，6k后用LPS刺激，收集转染后8，12，18k上清及细胞。细胞免疫组化法分析LPS刺激前后p65表达与分布变化，用酶联免疫吸附试验(ELISA)检测上清中肿瘤坏死因子—α(TNF—α)、白细胞介素—6(IL—6)蛋白表达，逆转录—聚合酶链反应(RT—PCR)法检测TNF—α、IL—6和IL—10mRNA转录。结果 4，8mg/L剂量哑铃形圈套0DNs可明显抑制LPS诱导的大鼠pMφTNF—α、IL—6转录和表达。结论 靶向NF—κB的哑铃形圈套0DNs具有拮抗炎症介质转录和表达的功能。     

LPS的直接诱导对肺微血管内皮细胞内NF-κB的影响

目的：探讨LPS的直接诱导作用对肺微血管内皮细胞(PMVEC)核因子κB(NF—κB)的影响。方法：100ng/ml LPS刺激PMVEC 0h、0．5h、1h、2h、4h、6h、8h或10ng/ml、50ng/ml、100ng/ml LPS刺激1h，凝胶电泳迁移率试验检测NF—κB的活化，免疫细胞化学观察其亚基p50、p65的核转位。并通过加入活化阻断剂TPCK观察对诱导的NF—κB活化的最响。结果：LPS的刺激迅速诱导p50、p65亚基核转位，活化NF—κB，1h达到简蜂，且呈剂量依赖关系，后逐渐下降。PDIC能显著抑制其活化，P＜0．01。结论：LPS的直接刺激诱导NF—κB的活化，这可能是LPS诱导炎症反应的一个重要环节。     

核因子-κB在内毒素刺激大鼠肺泡巨噬细胞中的动态变化及其在肿瘤坏死因子-α表达中的作用

目的　观察内毒素 (LPS)刺激对大鼠肺泡巨噬细胞 (alveolarmacrophages,AM)中核因子 -κB(NF -κB)活性的影响 ,探讨NF -κB在LPS刺激大鼠AM表达肿瘤坏死因子 -α(TNF -α)中的作用。　方法　体外观察LPS刺激大鼠AM中NF -κB活性的动态变化 ,应用圈套策略观察NF-κB在LPS刺激大鼠AM表达TNF -α中的作用。　结果　LPS刺激可使大鼠AM内NF -κB明显活化 ,并与LPS剂量正相关 ;抗体超迁移率实验结果显示p5 0、p6 5参与了NF -κB的活化 ;NF -κB的圈套硫代寡核苷酸 (S -ODN)能明显抑制LPS刺激大鼠AM表达TNF -α ,但不能完全阻断LPS诱导的TNF -α表达。　结论　NF -κB(p5 0 p6 5 )在LPS介导的炎症反应中发挥重要作用 ,LPS刺激大鼠AM表达TNF -α受NF -κB调控 ,但其他核转录因子可能也在TNF -α的表达中发挥重要作用。     

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.     

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.