MEBT/MEBO在慢性难愈合创面治疗中的基础与临床研究进展

烧伤创疡再生医疗技术(MEBT/MEBO)立足中西医结合理论,秉承中医整体观念,为创面营造生理性湿润环境的同时,通过转化生长因子β1(TGF-β1)/Smad3、细胞外信号调节激酶1/2(ERK1/2)等多种信号通路调控血管内皮生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)等多种细胞因子的表达,在糖尿病足、压...     

Flag-Tinagl1真核表达载体的构建及功能验证

目的 构建带Flag标签的肾小管间质性肾炎抗原样蛋白1(Tinagl1)基因真核表达载体,检测其对肝癌细胞HepG2 ERK/AKT磷酸化、生长和迁移的影响.方法 以人乳腺cDNA文库为模板,PCR扩增Tinagl1基因片段,将其插入pcDNA3.0载体,经双酶切和测序验证后,将重组质粒转染到肝癌HepG2细胞中,采用Western印迹检测重组蛋白对ERK和AKT磷酸化水平的影响,生长曲线和划痕实验检测其对肝癌细胞HepG2生长和迁移的影响.结果 双酶切和Western印迹结果表明,pcDNA3.0-Flag-Tinagl1真核表达质粒构建成功,Tinagl1可降低ERK和AKT的磷酸化水平;生长曲线和划痕实验表明Tinagl1抑制肝癌细胞HepG2生长和迁移.结论 构建了带Flag标签的Tinagl1真核表达载体,并能抑制肝癌细胞HepG2中ERK/AKT磷酸化、生长和迁移,为进一步研究Tinagl1在肝癌细胞中的功能奠定了基础.     

脱氧佛波醇乙酸酯通过激活PKC/ERK通路诱导急性髓系白血病细胞分化

目的研究脱氧佛波醇乙酸酯（12-deoxyphorbol 13-acetate,Prostratin,DPA）对人急性髓系白血病（AML）细胞株HL-60、NB4和U937细胞分化的影响,并证明其通过激活蛋白激酶C/胞外信号调节激酶（PKC/ERK）通路诱导细胞分化。方法 1μmol/L DPA作用HL-60细胞3 d后观察细胞形态的改变;不同剂量的DPA作用HL-60、NB4、U937细胞24 h后,流式细胞仪检测细胞分化表面标志变化;Western印迹检测ERK蛋白磷酸化的改变;流式细胞仪检测丝裂原活化蛋白激酶激酶（MEK）抑制剂和PKC抑制剂对细胞分化表面标志的影响;Western印迹检测MEK抑制剂和PKC抑制剂对ERK蛋白磷酸化的影响。结果 DPA可诱导HL-60细胞出现白血病细胞分化的形态。DPA剂量依赖性地诱导AML细胞CD11b表达。DPA在诱导分化剂量范围内可剂量依赖性地引起ERK磷酸化。MEK化学小分子抑制剂U0126可完全抑制DPA引起的ERK磷酸化和CD11b表达;PKC非选择性抑制剂GFX可完全抑制DPA引起的ERK磷酸化和CD11b表达。结论 DPA其通过激活PKC/ERK通路能够明显诱导白血病细胞分化。     

模拟失重条件下骨形态发生蛋白-2诱导的成骨细胞蛋白激酶MEK1活性的变化

目的观察模拟失重条件下由骨形态发生蛋白-2（BMP-2）诱导的大鼠骨肉瘤成骨样细胞（ROS17／2．8）中丝裂原激活的蛋白激酶／细胞外信号调节激酶的激酶1（MAPK／ERK kinase1，MEK1）活性的变化。方法在地面1G和回转器模拟失重条件下培养ROS17／2．8细胞，培养时间分别为24h、48h和72h。培养结束前1h加入BMP-2（500ng／ml），1h后提取细胞蛋白，应用Western Blotting法检测细胞中的总细胞外信号调节激酶1／2（总ERK1／2）和磷酸化细胞外信号调节激酶（p-ERK1／2）的含量。另设一空白对照组，即1G条件下不加BMP-2的培养组。结果7个实验组细胞的总ERK1／2蛋白表达量无明显差异；空白对照组p-ERK1／2的表达量极低，明显低于其他6组（P〈0．01）；各时间点模拟失重组p-ERK1／2表达量均低于1G对照组（P〈0．01）；随着失重时间延长p-ERK1／2表达量呈逐渐降低的趋势（P〈0．01）。结论模拟失重条件下BMP-2诱导的成骨细胞MAPK信号通路中蛋白激酶MEK1的活性下降。     

缺氧对肺腺癌细胞生长特性及血管形成的影响

目的探讨缺氧对肺腺癌肿瘤生长及血管形成的影响。方法人肺腺癌细胞株A549暴露于常氧(空气，5%CO2)、缺氧(1％O2，5％CO2，94%N2)、无氧(95％N2，5%CO2)环境48h后，将细胞接种于裸鼠皮下，观察其生长情况。通过免疫组化染色计数微血管密度，测定移植瘤组织中血管内皮细胞生长因子(VEGF)及碱性成纤维细胞生长因子(bFGF)的表达水平。结果移植10天后，缺氧组肿瘤体积显著大于常氧组。移植25天后缺氧组肿瘤体积、重量、微血管密度以及瘤组织中VEGF、bFGF水平均显著高于常氧组，而无氧组显著低于常氧组。结论适度缺氧可以刺激肺癌细胞VEGF、bFGF等生长因子的表达，促进移植瘤血管形成，提高肿瘤的生长能力；而严重缺氧损伤癌细胞，影响生长因子合成表达，阻碍血管形成，抑制肿瘤的生长。     

垂体腺瘤细胞VEGF表达与IL-1α和IL-6分泌的关系

目的：探讨HP75垂体腺瘤细胞株VEGF表达与细胞因子分泌的关系。方法：HP75细胞常规培养，采用ELISA方法分别检测VEGF、IL-1α和IL-6的水平。结果：（1）HP75细胞呈时间依赖性分泌VEGF、IL-1α和IL-6；（2）较大剂量的IL-1α呈剂量依赖性的刺激VEGF表达；（3）IL-6对VEGF的分泌无显著影响；（4）IL-1α和IL-6中和抗体对VEGF分泌无显著影响。结论：VEGF的表达与IL-1α的分泌密切相关，提示细胞因子在垂体腺瘤的生长过程中发挥一定作用。     

回转模拟失重对心肌成纤维细胞生长因子及ERK信号传导的影响

目的研究回转模拟失重条件下,心肌成纤维细胞中某些生长因子表达与分泌的变化,以及细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)信号通路的活化状态.方法通过新生大鼠原代培养获取心肌成纤维细胞,利用水平回转器模拟失重效应,对碱性成纤维细胞生长因子(bFGF)和转化生长因子β1(TGFβ1)的表达进行蛋白质印迹(western blotting)分析,以放射免疫检测方法测定细胞培养上清液中血管紧张素II(Ang II)浓度的变化;利用western blotting对ERK1/2及磷酸化ERK1/2分别进行检测并分析其活化状态. 结果与对照组比较,回转后心肌成纤维细胞的bFGF表达增加,TGFβ1表达没有明显变化,Ang II分泌增加;ERK1/2表达总量增加,但其磷酸化水平下降.结论不同的生长因子对回转模拟失重有不同的响应;作为生长因子的重要下游信号通路,ERK1/2的活化受到抑制.     

磷酸肌醇3-激酶在大鼠缺氧性肺动脉高压中的作用

目的：通过观察缺氧性肺动脉高压（HPH）大鼠肺组织中蛋白激酶B（AKT））和血管内皮生长因子（VEGF）的表达,研究磷酸肌醇3-激酶（PI3K）通路与VEGF的关系及其在HPH发病中的可能机制。方法：40只成年雄性Wistar大鼠随机分成对照组、低氧3、7、14、21d组,每组8只,测各组大鼠平均肺动脉压（mPAP）、右室肥大指数（RVHI）、血管形态学指标;免疫组化检测P-AKT和VEGF。结果：低氧7d后,mPAP开始上升[（23.53±1.78）mmHg],与对照组比较差异有显著性（P〈0.05）,低氧14d后达高水平并维持于此水平。肺血管重塑,RVIH改变在缺氧14d后出现。磷酸化AKT蛋白在对照组表达不明显,在低氧3d后表达上升,与对照组比较差异有显著性（P〈0.05）,且在低氧3、7、14、21d组肺血管内膜、中膜中,磷酸化AKT蛋白表达均为阳性。VEGF在低氧7d增高（0.188±0.018,P〈0.05）,14d达高峰（0.238±0.017,P〈0.05）。相关分析表明磷酸化AKT、VEGF蛋白、mPAP与血管重塑均正相关（P〈0.01）。磷酸化AKT与VEGF（内膜）成正相关（P〈0.01）。结论：磷酸化AKT与VEGF均在大鼠HPH的发病机制中发挥作用。     

垂体腺瘤细胞VEGF表达与细胞因子分泌的关系

目的:探讨垂体腺瘤细胞株HP75 VEGF表达与细胞因子IL-1α和IL-6分泌的关系。方法:HP75细胞常规培养,经各种因素处理后,取培养上清液,采用ELISA方法分别检测VEGF、IL-1α和IL-6的分泌水平。结果:①HP75细胞时间依赖性分泌VEGF、IL-1α和IL-6;②较大剂量的IL-1α剂量依赖性的刺激VEGF表达;③所用各种剂量的IL-6对VEGF的分泌均无显著影响;④IL-1α和IL-6中和抗体对VEGF基础分泌无显著影响。结论:VEGF的表达与IL-1α和IL-6的分泌密切相关,提示细胞因子在垂体腺瘤的生长过程中发挥重要作用。     

阿魏酸钠对抗谷氨酸诱导的鼠皮层神经元凋亡作用的影响

目的探讨MEK/ERK信号转导通路是否参与了阿魏酸钠（SF）对抗谷氨酸诱导的鼠皮层神经元凋亡作用。方法以谷氨酸诱导大鼠皮层神经元凋亡为模型。采用Westernblot观察Bcl-2、caspase-3、磷酸化ERK1/2表达的改变。结果阿魏酸钠能够显著降低谷氨酸诱导的神经细胞caspase-3的表达,提高Bcl-2、磷酸化ERK1/2的表达。PD98059可以减弱SF的保护作用。结论 MEK/ERK1/2通路参与阿魏酸钠对抗谷氨酸诱导的鼠皮层神经元凋亡。     

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.     

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.     

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）