TRAIL与kallistatin联合表达载体的构建及抗肿瘤活性分析

为研究肿瘤坏死因子相关凋亡诱导配体(TRAIL)与组织激肽释放酶结合蛋白(kallistatin)联合用药的抗肿瘤作用,构建TRAIL与kallistatin双表达的重组质粒pAM-CAG-Kal-IRES-TRAIL,将重组质粒转染A549,LO-2,NCI-H446和Hela细胞,考察其抗肿瘤活性.实验结果表明:构建的双表达载体能同时表达TRAIL与kallistatin,且均能分泌至培养基中;TRAIL与kallistatin联合表达对肿瘤细胞活力的抑制作用明显增强,诱导肿瘤凋亡的作用也明显增强,说明联合表达TRAIL与kallistatin能够增强抗肿瘤活性.     

田径运动肌肉损伤及康复对策研究

在田径教学、训练与比赛中,由于没有完全遵循人体运动的生理规律和调控生物节律,不能使机体的工作运行保持在良好的竞技状态,导致各种各样的运动创伤,严重地影响教学、训练与运动成绩的提高.文章从运动医学理论对田径运动中肌肉损伤的原因进行分析,寻找科学的田径运动损伤康复手段。     

Molecular dynamics and information on possible sites of interaction of intramyocellular metabolites in vivo from resolved dipolar couplings in localized H NMR spectra

Proton NMR resonances of the endogenous metabolites creatine and phosphocreatine ((P)Cr), taurine (Tau), and carnosine (Cs, β-alanyl-l-histidine) were studied with regard to residual dipolar couplings and molecular mobility. We present an analysis of the direct ¹H–¹H interaction that provides information on motional reorientation of subgroups in these molecules in vivo. For this purpose, localized ¹H NMR experiments were performed on m. gastrocnemius of healthy volunteers using a 1.5-T clinical whole-body MR scanner. We evaluated the observable dipolar coupling strength SD₀ (S = order parameter) of the (P)Cr-methyl triplet and the Tau-methylene doublet by means of the apparent line splitting. These were compared to the dipolar coupling strength of the (P)Cr-methylene doublet. In contrast to the aliphatic protons of (P)Cr and Tau, the aromatic H2 (δ = 8 ppm) and H4 (δ = 7 ppm) protons of the imidazole ring of Cs exhibit second-order spectra at 1.5 T. This effect is the consequence of incomplete transition from Zeeman to Paschen-Back regime and allows a determination of SD₀ from H2 and H4 of Cs as an alternative to evaluating the multiplet splitting which can be measured directly in high-resolution ¹H NMR spectra. Experimental data showed striking differences in the mobility of the metabolites when the dipolar coupling constant D₀ (calculated with the internuclear distance known from molecular geometry in the case of complete absence of molecular dynamics and motion) is used for comparison. The aliphatic signals involve very small order parameters S ≈ (1.4 − 3) × 10⁻⁴ indicating rapid reorientation of the corresponding subgroups in these metabolites. In contrast, analysis of the Cs resonances yielded S ≈ (113 − 137) × 10⁻⁴. Thus, the immobilization of the Cs imidazole ring owing to an anisotropic cellular substructure in human m. gastrocnemius is much more effective than for (P)Cr and Tau subgroups. Furthermore, ¹H NMR experiments on aqueous model solutions of histidine and N-acetyl-l-aspartate (NAA) enabled the assignment of an additional signal component at δ = 8 ppm of Cs in vivo to the amide group at the peptide bond. The visibility of this proton could result from hydrogen bonding which would agree with the anticipated stronger motional restriction of Cs. Referring to the observation that all dipolar-coupled multiplets resolved in localized in vivo ¹H NMR spectra of human m. gastrocnemius collapse simultaneously when the fibre structure is tilted towards the magic angle (θ ≈ 55°), a common model for molecular confinement in muscle tissue is proposed on the basis of an interaction of the studied metabolites with myocellular membrane phospholipids.     

基于cDNA宏阵列的系统聚类分析猪发育阶段的基因表达谱

取杜洛克猪胚胎第33,45,55，65．75天的背最长肌样本．用cDNA Macroarray分析方法和聚类分析技术分析了327个EST在骨骼肌内不同发育阶段的基因表达谱．结果表明有98条EST在不同发育时期显著差异表达．第33天和第45天两阶段基因表达状态相似．第55天和第65天基因表达状态相似．而第75天的基因表达与第55天和第65天两个阶段的基因表达具有相近的聚类关系．表达状态相近，基因功能相似的基因大都被聚类在一起．     

温度对荒漠沙蜥在体心脏单相动作电位的影响

采用吸附式电极记录了荒漠沙蜥在不同体温条件下 ,在体心脏的单相动作电位 (MAP:Monophasic Action Potential) .实验温度为 :5℃ ,10℃ ,15℃ ,2 0℃ ,2 5℃ ,30℃和 35℃ 7个温度等级 .结果表明 :(1)在最适温度 (2 0～ 2 5℃ )时 ,荒漠沙蜥心脏单相动作电位的基本波形与高等动物及人类基本相似 ;但也有其特殊性 :动作电位 0期除极期最大幅值为 11.4 2± 2 .5 8m V;复极 2期电位幅值基本保持恒定 ,呈平台期 (PP2 :Plateau Phase2 ) ;在复极 3期末 ,电位基本恢复到 0电位(膜电位为 0 m V) .(2 )随着体温的升高 (5～ 35℃ ) ,荒漠沙蜥心脏单相动作电位时程呈温度依赖性减小 ;0期最大除极速度 (Vmax)呈温度依赖性增加 (5℃为 77.0 8± 4 7.11m V/ms;35℃为 614.63±10 5 . 0 2 m V/ms) ;低于最适温度时 (2 0～ 2 5℃ ) ,在 2相平台期末 ,又出现一次除极电位 (后隆起波 ) .其最大电位为 0 .5 8± 0 .18m V.(3)随着体温的升高 ,荒漠沙蜥心脏单相动作电位复极 1期电位呈温度依赖性降低 (5℃为 2 .2 4± 1.31m V/ms;35℃为 0 .82± 0 .4 9m V/ms) .(4 )心电图 (ECG)的同步记录表明 :心电图中的 QRS波与心脏单相动作电位 0期除极完全相对应 ;T波与动作电位的复极 3期相对应 .     

急性冷暴露与持续冷驯化对中缅树鼩膈肌和心肌线粒体呼吸的影响

在急性冷驯化阶段(1h,4h,8h),中缅树膈肌线粒体状态IV呼吸与对照比较8小时组有显著的增强,心肌线粒体状态IV呼吸与对照比较在4小时和8小时有显著增强;在持续冷驯化阶段(1d,7d,14d,28d),中缅树膈肌和心肌线粒体状态IV呼吸与对照比较均有所增强,其中28天组增强十分显著。这一系列变化对于探讨中缅树膈肌与心肌在冷驯化中质子漏产热特征具有一定的意义。     

Vascular endothelial and smooth muscle cell culture on NaOH-treated poly(epsilon-caprolactone) films: a preliminary study for vascular graft development

Tissue engineering offers the potential of providing vessels that can be used to replace diseased and damaged native blood vessels. The endothelization of a synthetic vascular graft minimizes the failures associated with blood clotting and platelet activation. The aim of this study was to culture vascular-derived endothelial and smooth muscle cells on both untreated and NaOH-treated poly(epsilon-caprolactone) (PCL) films, a biocompatible and bio-resorbable polymer, and to evaluate the behavior of both cell types as a preliminary study for vascular graft development. PCL films were prepared by hot pressing; characterized by DSC, IR, SEM, and scanning force microscopy; and treated with NaOH to increase the surface hydrophilicity before cell culture. Endothelial and smooth muscle cells, isolated from pig cava vein, were characterized by immunofluorescence and confocal microscopy studies of endothelial nitric oxide synthase and alpha-smooth muscle actin. Good adhesion, growth, viability and morphology of both the endothelial and smooth muscle cells on PCL films were obtained, but a light stimulation of mitochondrial activity was observed during short culture times. NaOH treatment improved the adhesion and enhanced the proliferation in both cell types. This verified the possible use of this modified polymer as a support in the preparation of a synthetic vascular graft. [Diagram: see text] SEM micrograph of smooth muscle cells cultured on NaOH-treated PCL film. (Original magnification: 1000x).     

<Emphasis Type="Italic">L</Emphasis>-Carnitine Supplementation: A New Paradigm for its Role in Exercise

Summary. Early research investigating the effects of L-carnitine supplementation has examined its role in substrate metabolism and in acute exercise performance. These studies have yielded equivocal findings, partially due to difficulties in increasing muscle carnitine concentrations. However, recent studies have proposed that L-carnitine may play a different role in exercise physiology, and preliminary results have been encouraging. Current investigations have theorized that L-carnitine supplementation facilitates exercise recovery. Proposed mechanism is as follows: 1) increased serum carnitine concentration enhances capillary endothelial function; 2) increased blood flow and reduced hypoxia mitigate the cascade of ensuing, destructive chemical events following exercise; 3) thus allowing reduced structural damage of skeletal muscle mediated by more intact receptors in muscle needed for improved protein signaling. This paradigm explains decreased markers of purine catabolism, free radical formation, and muscle tissue disruption after resistance exercise and the increased repair of muscle proteins following long-term L-carnitine supplementation.     

A highly sensitive method for quantification of myosin light chain phosphorylation by capillary isoelectric focusing with laser-induced fluorescence detection

Activation of myosin II by phosphorylation of the 20 kDa regulatory light chains (LC20) has been implicated in numerous contractile and motile events, e.g., smooth muscle contraction, cytokinesis, and cell migration. The ability to analyze LC20 phosphorylation in minute samples is critical to determine the importance of LC20 phosphorylation in diverse physiological processes. We have developed a method for the separation and quantification of unphosphorylated, monophosphorylated, and diphosphorylated LC20 with a detection limit of 1 pg (50 amol). LC20 is initially isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transblotted to a polyvinlyidene difluoride (PVDF) membrane. The region of the membrane containing the LC20 band (identified by electrophoresis of purified LC20 in a neighboring lane) is cut out and fluorescently labeled with Alexa Fluor 488 C5 maleimide. The labeled LC20 is eluted from the membrane with detergent and subjected to capillary isoelectric focusing (CIEF) to separate unphosphorylated, mono-, and diphosphorylated LC20, which are detected and quantified by laser-induced fluorescence (LIF). A linear relationship between log(peak area) and log(LC20 amount) is observed over the range of 50 amol-150 fmol. Quantification of LC20 phosphorylation by CIEF with LIF detection was compared with three commonly used methods with much lower levels of sensitivity: urea/glycerol-PAGE with Western blotting, phosphorylation by [gamma-32P]ATP with Cerenkov counting, and phosphorylation by [gamma-32P]ATP followed by SDS-PAGE, autoradiography, and scanning densitometry. All four methods gave very similar quantitative results, the major difference being that the new method exhibits 3000-fold enhanced sensitivity. This method is therefore applicable to quantitative analysis of phosphorylation of minute quantities of LC20.     

海水养殖鱼类肌肉中微量元素的测定

应用原子吸收光谱法测定了湛江海域十种养殖鱼类肌肉中微量元素的含量。结果表明,海水养殖鱼肌肉中含有丰富的K、Na、Mg、Ca、Zn、Fe等微量元素,经常食用,可以提高机体的免疫力,增强体质,能较好地满足人体对微量元素的需要。