运动性疲劳的线粒体膜分子机理研究V:线粒体CoQ和质子循环共同参与ROS循环--运动模型中的证据

目的 :以运动、外源性补充辅酶Q (coenzymeQ ,CoQ)以及运动合并补充CoQ作为干预手段 ,观察线粒体CoQ结合含量、质子跨膜势能与活性氧产生之间的相互关系 ,进一步探讨运动性内源活性氧生成和代谢的线粒体膜分子机制。方法 :雄性SD大鼠 36只 ,随机分为 :(1)安静对照组 (R ,n =6 ) ;(2 )运动中对照组 (Em ,n =6 ) ;(3)力竭对照组 (Ei,n =6 ) ;(4 )安静补药组 (QR ,n=6 ) ;(5 )运动中补药组 (Qm ,n =6 ) ;(6 )力竭补药组 (Qi ,n =6 )。采用三级递增负荷力竭运动模型 ,测定心肌线粒体CoQ9及CoQ10 结合含量、ROS产生速率以及线粒体膜质子泵出与电子传递比值(H+/ 2e)。结果 :(1)Em和Qm组心肌线粒体ROS产生、H+/ 2e和CoQ结合含量分别比R和QR组显著增高 ,并且 ,Qm组三项指标显著高于Em组。 (2 )相关分析表明 ,CoQ结合含量分别与H+/ 2e和ROS产生速率呈线性正相关。结论 :以外源性补充CoQ10 和 /或运动应激作为干预手段时 ,心肌线粒体CoQ含量升高导致建立高质子跨膜势能 ,并进而增加活性氧生成 ,进一步支持“活性氧循环”与Q循环和质子循环并存和共同运转可能是运动性内源活性氧生成及代谢的重要机制     

Circulating microparticle tissue factor,thromboembolism and survival in pancreaticobiliary cancers

Background

Tissue factor (TF), the physiologic initiator of coagulation, is over-expressed in pancreatic cancer, and is associated with a pro-coagulant and pro-angiogenic state. We hypothesized that in patients with pancreaticobiliary cancers (PBC), elevated circulating microparticle-associated TF (MP-TF) activity would be associated with thrombosis and worsened survival.

Patients and Methods

Clinical data and plasma were obtained for consecutive patients with PBC seen at Roswell Park Cancer Institute from 2005-08. MP-TF activity levels were measured using a TF-dependent FXa generation assay.

Results

The study population comprised 117 patients, including pancreatic (n = 80), biliary (n = 34) or unknown primary histologically consistent with PBC (n = 3). Of these, 52 patients (44.5%) experienced thromboembolism, including pulmonary embolism (n = 15), deep venous thrombosis (n = 21) and other arterial or venous events (n = 32). Mean TF was 2.15 (range 0.17- 31.01) pg/mL. Median survival was 98.5 days for MP-TF activity ≥ 2.5 pg/mL versus 231 days for MP-TF activity < 2.5 pg/mL (p < 0.0001). In multivariate analysis, elevated MP-TF activity was associated with both VTE (OR 1.4, 95% CI 1.1-1.6) and mortality (HR 2.5, 95% CI 1.4-4.5).

Conclusions

Elevated circulating MP-TF activity is associated with thrombosis and worsened survival in patients with PBC. MP-TF activity as a prognostic biomarker warrants further prospective evaluation.     

骨创伤修复基础研究的若干新进展

分子生物学技术的迅猛发展给骨创伤后组织修复的基础研究和临床治疗提供了新的思路和手段。本文概述近年来在生长因子、基因治疗以及组织工程三方面进展对骨创伤后组织修复基础研究和临床治疗的促进作用。转化生长因子(TGF-β)、骨形成蛋白(BMP)、成纤维细胞生长因子(FGF）、血小板源性生长因子(PDGF）和胰岛素样生长因子(IGF)在骨修复中起重要作用，基因治疗将在软组织修复、骨形成和神经再生方面发挥作用。组织工程化骨与软骨、皮肤、血管等研究也将对骨科创伤修复产生积做影响。     

糖尿病雄性生殖损伤及其发生机制

糖尿病是由多种病因引起的代谢性疾病 ,其生殖损伤严重危害机体的健康。本文综述了糖尿病雄性生殖损伤病理生理改变及其发生机制 ,高血糖导致的氧化应激在糖尿病雄性生殖损伤中的重要作用 ,以及某些抗氧化剂可能成为有效治疗糖尿病生殖损伤的药物。     

人IP-10启动子的克隆和转录活性的检测

目的克隆干扰素诱导蛋白10(IP-10)5′非编码区(NCR)片段,检测克隆的IP-10启动子驱动的荧光素酶报告基因在LPS激活的人脐静脉内皮细胞(HUVEC)中的转录活性。方法提取人外周血淋巴细胞基因组DNA,以其为模板,利用巢式PCR扩增人IP-10启动子片段,测序正确后克隆入荧光素酶报告基因表达载体pGL3;将重组质粒pGL3/IP-10P导入HUVEC细胞,检测LPS刺激下荧光素酶活性。结果正确克隆出人IP-10启动子(-2028~-1)片段,成功构建了pGL3/IP-10P报告基因表达载体;证实了LPS可诱导HUVEC细胞中IP-10的高表达。结论成功克隆出人的IP-10启动子片段,并利用荧光素报告基因证实了在HUVEC细胞中LPS可诱导IP-10高转录表达,为深入研究LPS诱导IP-10转录表达的调控机制提供了基础。     

贫铀弹伤害及其医学防护

贫铀在军事上主要用于加固装甲和制造贫铀弹。贫铀武器多次在实战中被使用,造成人员死伤,装备破坏,环境污染,产生了严重后果。本从贫铀弹的4种杀伤因素、贫铀弹伤害的主要临床病理特点、医学防护和救治几个方面阐述了贫铀弹所致的伤害及其医学防护,同时也对防护中几个基本问题提出了自己的看法。     

利用PCR方法获得shRNA抑制内外源性基因的表达

目的探讨PCR法获得的shRNA对外源性基因和内源性基因表达的抑制效果。方法选择绿色荧光蛋白(GFP)和大鼠肾脏系膜细胞高丰度表达的纤维连接蛋白(FN)分别作为外源性和内源性基因的靶基因,设计特异性引物,利用PCR扩增获得shGFPRNA和shFNRNA。前者与pEGFP质粒共转染至293T细胞,48h后进行激光共聚焦检测细胞表达GFP阳性率;后者转染至大鼠系膜细胞(RMC)中,48h后提取总RNA逆转录成cDNA,经realtimeRTPCR检测FN的mRNA表达水平,提取细胞总蛋白进行Westernblot检测FN蛋白表达水平。结果PCR获得的shGFPRNA产物能有效抑制外源性基因GFP的表达,抑制效率可达70%±3.2%;shRNA转染组内源性基因FN的表达水平明显下降,与对照组相比有显著性差异(P<0.05)。结论PCR方法可快速简单、廉价地获得shRNA,对内源性或外源性基因进行特异性抑制。     

Prophylactic treatment of very premature infants with human surfactant

We undertook a randomized, controlled trial to determine whether human surfactant administered endotracheally at birth to very premature infants (gestational age, 24 to 29 weeks) would prevent the respiratory distress syndrome or reduce its severity. Thirty-one treated infants (birth weight, 938 +/- 286 g) were compared in a blinded fashion with 29 control infants (birth weight, 964 +/- 174 g). The lecithin/sphingomyelin ratio was less than 2 in all infants, and phosphatidylglycerol was not present in amniotic fluid or tracheal fluids at birth, indicating a deficiency of surfactant in the lungs. The principal dependent variables were neonatal death, the incidence of bronchopulmonary dysplasia, and the infant's requirement for respiratory support (and its complications). The surfactant-treated group had significantly fewer deaths than the control group (16 percent vs. 52 percent, P less than 0.001), fewer cases of bronchopulmonary dysplasia (16 percent vs. 31 percent), and significantly fewer cases of pulmonary interstitial emphysema (P less than 0.001) and pneumothorax (P less than 0.02). Prophylactic treatment with human surfactant also substantially reduced the period of neonatal intensive care. We conclude that treatment with human surfactant offers promise for improving the survival of very premature infants with a surfactant deficiency and for reducing the pulmonary sequelae of the respiratory distress syndrome.     

新型定位板在经椎间孔腰骶神经根封闭术术前定位中的应用

目的 :探讨新型定位板应用在腰椎后外侧经椎间孔腰骶神经根封闭术的术前定位中的有效性、可靠性。方法:2015年3月~2016年3月收治腰椎退行性疾病患者102例,其中单节段腰椎间盘突出症51例,单节段腰椎管狭窄症42例,经皮内窥镜下腰椎间盘切除术后症状复发9例,采用随机数字表随机分入两组后行腰椎后外侧经椎间孔腰骶神经根封闭术。A组54例,手术节段为L3/4 8例、L4/5 28例、L5/S1 18例,采用新型定位板术前定位;B组48例,手术节段为L3/4 8例、L4/5 26例、L5/S1 14例,采用金属定位针术前定位。两组患者年龄、性别、手术节段、保守治疗时间均无统计学差异(P0.05)。记录两组术前的透视次数和准备时间、穿刺时间、穿刺期透视次数、手术并发症、穿刺术后1h穿刺区疼痛VAS评分,并进行统计学分析。结果:A组术前的准备时间为5.2±1.0min、透视次数为1.1±0.3次,穿刺时间9.6±2.2min,穿刺期透视次数3.1±1.0次;B组术前的准备时间为10.7±2.3min、透视次数为3.8±1.2次,穿刺时间16.3±3.3min,穿刺期透视次数4.6±0.6次,两组比较均有统计学差异(P0.05),A组均优于B组。两组均未出现椎管内血肿、腹腔脏器损伤、下肢感觉和运动功能异常,B组出现硬膜刺裂1例(1/48)、穿刺区域皮下血肿4例(4/48),两组并发症发生率无统计学差异(P0.05);术后1h穿刺区域疼痛VAS评分,A组为3.4±0.5分,B组为5.0±0.9分,有统计学差异(P0.05)。结论:对于经椎间孔腰骶神经根封闭术,使用新型定位板术前定位,并进行穿刺路径设计,可减少术前透视次数、术前准备时间,有助于缩短穿刺时间及穿刺期透视次数,定位板具有使用方便、可靠、有效等优点。