燃煤污染型氟中毒患者氟暴露、骨相损伤程度与骨形成标志物的关系

慢性氟暴露是世界范围内的公共卫生问题之一.为研究燃煤污染型氟中毒患者中氟暴露、骨相损伤程度与骨形成标志物血清碱性磷酸酶(ALP)、骨钙素(BGP)之间的相关关系,以贵州省织金县荷花村(燃煤污染型地方性氟中毒病区)和安顺市张官村(非氟暴露村)为调查点,采集环境样品,采用氟离子选择电极法测定环境介质及食物中的氟含量.在知情同意的原则下,对295例氟暴露和85例非氟暴露调查对象进行氟斑牙、氟骨症诊断;收集其尿液及外周血,测定尿氟(UF)浓度、ALP活性和BGP含量.结果表明: 病区环境中大米、辣椒、玉米、饮水、黏土、菜土、煤以及室内外空气氟含量均明显高于对照区,但较以往报道数据降低;随着尿氟浓度的升高,ALP活性、BGP含量显著升高,氟骨症病情差异有统计学意义,氟斑牙病情差异无统计学意义;氟骨症病情与ALP活性、BGP含量呈正相关.表明燃煤污染型低氟暴露可引起人群的骨相损害,且ALP、BGP可用于评估氟骨症患者骨转换情况,在氟骨症的辅助诊断、疗效评估中有着一定的应用价值.     

PAX3 Confers Functional Heterogeneity in Skeletal Muscle Stem Cell Responses to Environmental Stress

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Purification and characterization of cytosolic glyceraldehyde-3-phosphate dehydrogenase from the dromedary camel

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (EC 1.2.1.12),a key enzyme ofcarbon metabolism,was purified and characterized to homogeneity from skeletal muscle of Camelusdromedarius.The protein was purified approximately 26.8 folds by conventional ammonium sulphatefractionation followed by Blue Sepharose CL-6B chromatography,and its physical and kinetic propertieswere investigated.The native protein is a homotetramer with an apparent molecular weight of approximately146 kDa.Isoelectric focusing analysis showed the presence of only one GAPDH isoform with an isoelectricpoint of 7.2.The optimum pH of the purified enzyme was 7.8.Studies on the effect of temperature onenzyme activity revealed an optimal value of approximately 28-32 ℃ with activation energy of 4.9 kcal/mol.The apparent K_m values for NAD~ and DL-glyceraldehyde-3-phophate were estimated to be 0.025±0.040mM and 0.21±0.08 mM, respectively. The V_(max) of the purified protein was estimated to be 52.7±5.9 U/mg.These kinetic parameter values were different from those described previously, reflecting protein differencesbetween species.     

Heat shock pretreatment enhances porcine myoblasts survival after autotransplantation in intact skeletal muscle

Myoblast transplantation (MT) is a cell-based gene therapy treatment, representing a potential treat-ment for Duchenne muscular dystrophy (DMD), cardiac failure and muscle trauma. The rapid and mas-sive death of transplanted cells after MT is considered as a major hurdle which limits the efficacy of MT treatment. Heat shock proteins (HSPs) are overexpressed when cells undergo various insults. HSPs have been described to protect cells in vivo and in vitro against diverse insults. The aim of our study is to investigate whether HSP overexpression could increase myoblast survival after autotransplantation in pig intact skeletal muscle. HSP expression was induced by warming the cells at 42℃ for 1 h. HSP70 expression was quantified by Western blot and flow cytometry 24 h after the treatment. To investigate the myogenic characteristics of myoblasts, desmin and CD56 were analysed by Western blot and flow cytometry; and the fusion index was measured. We also quantified cell survival after autologous transplantation in pig intact skeletal muscle and followed cell integration. Results showed that heat shock treatment of myoblasts induced a significative overexpression of the HSP70 (P < 0.01) without loss of their myogenic characteristics as assessed by FACS and fusion index. In vivo (n=7), the myoblast survival rate was not significantly different at 24 h between heat shock treated and non- treated cells (67.69% ± 8.35% versus 58.79% ± 8.35%, P > 0.05). However, the myoblast survival rate in the heat shocked cells increased by twofold at 48 h (53.32% ± 8.22% versus 28.27% ± 6.32%, P < 0.01) and more than threefold at 120 h (26.33% ± 5.54% versus 8.79% ± 2.51%, P < 0.01). Histological analy-sis showed the presence of non-heat shocked and heat shocked donor myoblasts fused with host myoblasts. These results suggested that heat shock pretreatment increased the HSP70 expression in porcine myoblasts, and improved the survival rate after autologous transplantation. Therefore, heat shock pretreatment of myoblast in vitro is a simple and effective way to enhance cell survival after transplantation in pig. It might represent a potential method to overcome the limitations of MT treat-ment.     

不同运动方式对2型糖尿病大鼠骨骼肌Rab5蛋白及糖代谢的影响*

目的: 探讨持续运动和间歇负重运动对2型糖尿病(T2DM)骨骼肌组织细胞形态、骨骼肌Rab5 mRNA及蛋白表达、骨骼肌糖代谢的影响。方法: SD大鼠选取8只为空白对照组(CR),其他采用高脂高糖饲料喂养6周后,腹腔注射STZ(35 mg/kg)构建T2DM模型。选取24只T2DM分3组(n=8),分别为:T2DM模型组(DRM)、持续运动组(DCRE)、间歇负重运动组(DWRE)。持续运动方案:为前1～2 周准备活动15 m/min(10 min)、运动20 m/min(40 min)、整理活动15 m/min(10 min),后3～8周为 18 m/min(10 min)、25 m/min(40 min)、15 m/min(10 min);间歇负重运动方案:采用负荷重量为15%(1～2周)、30%(3～4周)、45%(5～8周),运动均为15 m/min(5 min),共12组,组间休息3 min。8周后,通过HE观察骨骼肌病理形态变化,qRT-PCR检测骨骼肌Rab5、葡萄糖转运酶4(GLUT4)的mRNA表达,免疫荧光组化技术及Western blot检测骨骼肌Rab5的蛋白表达,ELISA检测血浆Rab5和糖化血红蛋白(GHb)浓度。结果: 相比CR,DRM存在骨骼肌病理损伤,骨骼肌Rab5mRNA及蛋白表达、GLUT4 mRNA表达均降低(P＜0.01),血浆Rab5和GHb均显著升高(P＜0.01);与DRM比较, DCRE、DWRE骨骼肌病理损伤均显著减轻,骨骼肌Rab5 mRNA及蛋白表达、GLUT4 mRNA表达均升高(P＜0.05,P＜0.01),血浆Rab5和GHb降低(P＜0.01);DCRE与DWRE组间均无统计学差异(P＞0.05)。结论: 2种运动方式均能改善2型糖尿病大鼠骨骼肌病理损伤,并可通过提高骨骼肌Rab5基因和蛋白表达从而增强 GLUT4转运能力,缓解骨骼肌糖代谢稳态失衡,但2种运动方式对骨骼肌Rab5蛋白和糖代谢的影响无明显差异性。     

Mechanical loading stimulates hypertrophy in tissue-engineered skeletal muscle: Molecular and phenotypic responses

Mechanical loading of skeletal muscle results in molecular and phenotypic adaptations typified by enhanced muscle size. Studies on humans are limited by the need for repeated sampling, and studies on animals have methodological and ethical limitations. In this investigation, three-dimensional skeletal muscle was tissue-engineered utilizing the murine cell line C2C12, which bears resemblance to native tissue and benefits from the advantages of conventional in vitro experiments. The work aimed to determine if mechanical loading induced an anabolic hypertrophic response, akin to that described in vivo after mechanical loading in the form of resistance exercise. Specifically, we temporally investigated candidate gene expression and Akt-mechanistic target of rapamycin 1 signalling along with myotube growth and tissue function. Mechanical loading (construct length increase of 15%) significantly increased insulin-like growth factor-1 and MMP-2 messenger RNA expression 21 hr after overload, and the levels of the atrophic gene MAFbx were significantly downregulated 45 hr after mechanical overload. In addition, p70S6 kinase and 4EBP-1 phosphorylation were upregulated immediately after mechanical overload. Maximal contractile force was augmented 45 hr after load with a 265% increase in force, alongside significant hypertrophy of the myotubes within the engineered muscle. Overall, mechanical loading of tissue-engineered skeletal muscle induced hypertrophy and improved force production.     

Estimating fossil biomass from skeletal mass in marine invertebrates

Palaeoecology uses the numerical abundance and the occurrence of species to evaluate the dynamics of past communities, but biomass – the quantity of soft tissue – is the critical currency needed to capture the flow and role of nutrients in modern ecosystems. Acquiring biomass data from fossil assemblages has, however, remained challenging, thus limiting the analysis of net secondary production in palaeocommunities. Prior models relate shell size or shell biovolume to fossil biomass. These models neglect shell fragments and, moreover, use units of biovolume (cm³) that are not directly related to those of biomass (g), making the models difficult to tune and the coefficients highly specific. To remedy these shortcomings, I evaluate skeletal mass as a means of estimating the soft tissue biomass of fossil taxa, using ratios among biomass, skeletal mass and the total wet mass of living representatives of extant species, so that skeletal mass alone can be used to estimate grams of organic biomass. Data on total wet mass, organic carbon mass, and shell mass were acquired from more than 80 live‐collected individuals from eight families in three major, shelly macrobenthic groups (Mollusca, Brachiopoda, Arthropoda) and supplemented with counterpart data from the literature to increase taxonomic breadth. This new shell‐mass model provides more accurate and precise biomass estimates than models based on the linear dimensions of shells, expanding our ability to examine the interplay between organisms and their environments.