MiRNA-199a-3p Regulates C2C12 Myoblast Differentiation through IGF-1/AKT/mTOR Signal Pathway

MicroRNAs constitute a class of ~22-nucleotide non-coding RNAs. They modulate gene expression by associating with the 3′ untranslated regions (3′ UTRs) of messenger RNAs (mRNAs). Although multiple miRNAs are known to be regulated during myoblast differentiation, their individual roles in muscle development are still not fully understood. In this study, we showed that miR-199a-3p was highly expressed in skeletal muscle and was induced during C2C12 myoblasts differentiation. We also identified and confirmed several genes of the IGF-1/AKT/mTOR signal pathway, including IGF-1, mTOR, and RPS6KA6, as important cellular targets of miR-199a-3p in myoblasts. Overexpression of miR-199a-3p partially blocked C2C12 myoblast differentiation and the activation of AKT/mTOR signal pathway, while interference of miR-199a-3p by antisense oligonucleotides promoted C2C12 differentiation and myotube hypertrophy. Thus, our studies have established miR-199a-3p as a potential regulator of myogenesis through the suppression of IGF-1/AKT/mTOR signal pathway.     

骨髓间充质干细胞对大鼠脑胶质瘤C6细胞系分化的影响

目的探讨骨髓间充质干细胞(Bone mesenchymal stem cells,BMSCs)对大鼠脑胶质瘤C6细胞系分化的影响及其相关机制。方法用Transwell小室共培养BMSCs与C6细胞,采用细胞计数法检测C6细胞增殖水平;流式细胞术检测C6细胞的细胞周期;免疫荧光与免疫组化法分别检测波形蛋白(vimentin)和胶质纤维酸性蛋白(Glia lfibrillary acidic protein,GFAP)的表达;生化法检测谷氨酰胺合成酶(Glutamine synthetase,GS)的活性。结果 BMSCs对C6细胞的增殖具有抑制作用;与BMSCs进行双层培养72h的C6细胞出现G0/G1期细胞周期阻滞,双层培养组的C6细胞GFAP表达明显增强,同时vimentin表达减弱,并且这两种分化标志物在细胞内的分布及排列发生了改变;双层培养组C6细胞内GS活性升高。结论 BMSCs能诱导C6细胞向成熟星形胶质细胞分化,可能与BMSCs分泌的某些细胞因子有关。     

MicroRNA-27a Is Induced by Leucine and Contributes to Leucine-Induced Proliferation Promotion in C2C12 Cells

Leucine, a branched chain amino acid, is well known to stimulate protein synthesis in skeletal muscle. However, the role of leucine in myoblast proliferation remains unclear. In this study, we found that leucine could promote proliferation of C2C12 cells. Moreover, expressions of miR-27a and myostatin (a bona fide target of miR-27a) were upregulated and downregulated, respectively, following leucine treatment. We also found that miR-27a loss-of-function by transfection of a miR-27a inhibitor suppressed the promotion of myoblast proliferation caused by leucine. Our results suggest that miR-27a is induced by leucine and contributes to leucine-induced proliferation promotion of myoblast.     

CLA Reduces Inflammatory Mediators from A427 Human Lung Cancer Cells and A427 Conditioned Medium Promotes Differentiation of C2C12 Murine Muscle Cells

Conjugated linoleic acid (CLA) is thought to have anti-proliferative and anti-inflammatory properties, but its effect on cancer cachexia is unknown. Two effects were here investigated: that of CLA on inflammatory mediator production in human lung cancer cells, and that of reduced mediators on the myogenic differentiation of murine muscle C2C12 cells. The latter cells were grown in medium conditioned by human lung cancer A427 cells, with or without CLA, to mimic only the effect of molecules released from the tumor “in vivo”, excluding the effect of host-produced cachectic factors. The results obtained show that CLA was found to reduce the production of tumor necrosis factor-α, interleukin (IL)-1β and prostaglandin E2 (PGE2), but had no effect on IL-6 production. The mechanisms underlying the effect of CLA on cytokine or PGE2 release in A427 cells are probably mediated by activation of peroxisome proliferator-activated receptor (PPAR)α, which increased at 24 h CLA treatment. In turn, the reduced content of inflammatory mediators in medium conditioned by A427 cells, in the presence of CLA, allowed muscle cells to proliferate, again by inducing PPAR. The involvement of PPARα was demonstrated by treatment with the antagonist MK-886. The findings demonstrate the anti-inflammatory and myogenic action of CLA and point to its possible application as a novel dietary supplement and therapeutic agent in inflammatory disease states, such as cachexia.     

FAHFAs Regulate the Proliferation of C2C12 Myoblasts and Induce a Shift toward a More Oxidative Phenotype in Mouse Skeletal Muscle

Branched fatty acid esters of hydroxy fatty acids (FAHFAs) are endogenous lipids reported to have antidiabetic and anti-inflammatory effects. Since skeletal muscle is a major target for insulin, the aim of this study is to explore for the first time the influence of several FAHFAs in C2C12 myoblasts and in skeletal muscle phenotype in mice. Here, we show that eleven FAHFAs belonging to different families inhibit C2C12 myoblast proliferation. In addition, all FAHFAs decreased mitochondrial cytochrome c oxidase activity without affecting reactive oxygen species production and the mitochondrial network. During C2C12 myoblasts differentiation, we found that two of the most active lipids, 9-PAHPA and 9-OAHPA, did not significantly affect the fusion index and the expression of myosin heavy chains. However, we found that three months’ intake of 9-PAHPA or 9-OAHPA in mice increased the expression of more oxidative myosin in skeletal muscle without affecting skeletal muscle mass, number, and mean fiber area, mitochondrial activity, and oxidative stress parameters. In conclusion, our study indicated that the eleven FAHFAs tested decreased the proliferation rate of C2C12 myoblasts, probably through the inhibition of mitochondrial activity. In addition, we found that 9-PAHPA or 9-OAHPA supplementation in mice induced a switch toward a more oxidative contractile phenotype of skeletal muscle. These data suggest that the increase in insulin sensitivity previously described for these two FAHFAs is of muscular origin.     

葡萄糖的含量对LiFePO4性能的影响

以葡萄糖为表面活性剂,采用水热法制备LiFePO4正极材料,并且考察了葡萄糖加入量对LiFePO4正极材料性能的影响。用XRD、SEM及电池测试系统对LiFePO4材料进行了研究。结果表明:葡萄糖不仅能够有效的对颗粒进行分散;而且还能够在高温高压下,裂解为碳,提高了材料的比容量;当葡萄糖含量为4%时在0.1C倍率下放电容量达到129.7mAh/g,体现良好的倍率放电特性。     

Rosiglitazone,but Not Epigallocatechin‐3‐Gallate,Attenuates the Decrease in PGC‐1α Protein Levels in Palmitate‐Induced Insulin‐Resistant C2C12 Cells

Alteration of lipid metabolism is an important mechanism for the treatment of insulin resistance. PGC‐1α, a key regulator of mitochondrial biogenesis and function, plays an important role in the improvement of insulin sensitivity by increasing fatty acids β‐oxidation. In the present study, the effects of epigallocatechin‐3‐gallate (EGCG), an anti‐obesity agent and enhancer of lipid catabolism, on PGC‐1α protein expression was examined and compared with anti‐diabetic drug rosiglitazone (RGZ). After differentiation of C2C12 myoblasts to myotubes, insulin resistance was induced by palmitate treatment. Then the expression of the PGC‐1a gene and glucose uptake were evaluated before and after treatment with RGZ and EGCG. Palmitate treatment significantly decreased PGC‐1α protein expression in C2C12 cells (P < 0.05). RGZ could restore the expression of PGC‐1α in palmitate treated cells (P > 0.05), while EGCG had no significant effect on the expression of this gene (P < 0.05). RGZ and EGCG significantly improved glucose uptake (by 2‐ and 1.54‐fold, respectively) in myotubes treated with palmitate. These data suggest that RGZ and EGCG both exert their anti‐diabetic activity by increasing insulin sensitivity, but with different molecular mechanisms. This effect of RGZ, unlike EGCG, is mediated, at least partly, by increasing PGC‐1α protein expression.     

Tceal5 and Tceal7 Function in C2C12 Myogenic Differentiation via Exosomes in Fetal Bovine Serum

The proliferation and differentiation of skeletal muscle cells are usually controlled by serum components. Myogenic differentiation is induced by a reduction of serum components in vitro. It has been recently reported that serum contains not only various growth factors with specific actions on the proliferation and differentiation of myogenic cells, but also exogenous exosomes, the function of which is poorly understood in myogenesis. We have found that exosomes in fetal bovine serum are capable of exerting an inhibitive effect on the differentiation of C2C12 myogenic cells in vitro. In this process of inhibition, the downregulation of Tceal5 and Tceal7 genes was observed. Expression of these genes is specifically increased in direct proportion to myogenic differentiation. Loss- or gain- of function studies with Tceal5 and Tceal7 indicated that they have the potential to regulate myogenic differentiation via exosomes in fetal bovine serum.     

2,10,12-三甲基-6-氧-12H-双苯并基[d,g][1,3,2]-二氧硫杂八环合成和表征

以三乙胺为缚酸剂,1,1-双(2-羟基-5-甲基苯基)乙烷和二氯亚砜为原料,合成了双酚的亚硫酸酯的化合物2,10,12-三甲基-6-氧-12H-双苯并[d,g][1,3,2]-二氧硫杂八环,最佳合成的物质的量比为:双酚∶二氯亚砜∶三乙胺=1∶2.0∶2.2,反应时间为2 h,产品收率为72%(以双酚计)。标题化合物经元素分析、IR、1H NMR对产物进行了结构鉴定,并用X射线单晶衍射法确定其单晶结构。     

Boosting C2H6/C2H4 separation in scalable metal-organic frameworks through pore engineering

The development of ethane (C₂H₆)-selective adsorbents for ethylene (C₂H₄) purification, although challenging, is of prime industrial importance. Pillared-layer metal-organic frameworks (MOFs) possess facilely tunable pore structure and functionality, which means they have excellent potential for high-performance C₂H₆/C₂H₄ separation applications. Herein, we report a family of isostructural pillared-layer MOFs with various metal centers M and co-ligands L, M₂(D-cam)₄L₂ (denoted M-cam-L; M = Cu, Co, Ni; L = pyz, apyz, dabco), with a variety of pore surface properties. All of the M-cam-L materials exhibit preferential adsorption for C₂H₆ over C₂H₄. In particular, Ni-cam-pyz exhibits the highest C₂H₆ capture capacity (68.75 cm³ g⁻¹ at 1 bar and 298 K), Cu-cam-dabco possesses the greatest C₂H₆/C₂H₄ adsorption selectivity (2.3), and the lowest isosteric heat of adsorption is demonstrated for Cu-cam-pyz (20.1 kJ mol⁻¹). Dynamic column breakthrough experiments also confirmed the excellent separation performance of M-cam-pyz and M-cam-dabco materials. The synthesis route of the M-cam-L materials is easily scaled-up under laboratory conditions, and hence this class of MOFs is promising for practical C₂H₄ purification.     

乙烷-氧-水氧化裂解制乙烯反应的研究

研究了不同反应体系组成的乙烷造反然的反应性能,考究了乙烷－氧－水反应体系氧化裂解制乙烯的反应条件。结果表明,在不同反应体系中,以Ｃ２Ｈ６－Ｏ２－Ｈ２Ｏ氧化裂解制乙烯反应性能最佳,８００℃的乙烷转化率为８５．１％,乙烯选择性为６８．１％,乙烯收率可达５８％,Ｃ２Ｈ６－Ｏ２－Ｈ２Ｏ氧化裂解帛乙烯体系最佳工艺参数;反应温度为８５０℃,原料气组成为５０．５％,Ｃ２Ｈ６＋２５．２％Ｏ２＋２４．３％Ｈ２Ｏ停留     

一元和三元型含水硫铝酸钙热力学研究

根据温元凯算法的主体思想,计算了一元和三元型硫铝酸钙的Gibbs自由能和其反应Gibbs自由能分别为G3oCaO.Al2O3.CaSO4.12H2O=-8 984.805 kJ/mol和G3oCaO.Al2O3.3CaSO4.6H2O=-10 076.685 kJ/mol。铝酸钠溶液脱硫反应的Gibbs自由能△G1o=-195.265 kJ/mol,△G2o=-187.725 kJ/mol,两式中△Gor<<0。由计算结果可知,有望实现以生成三元型硫铝酸钙的形式脱硫,使脱硫的效率提高3倍,且降低Al2O3损失。     

miR-378a-3p Participates in Metformin’s Mechanism of Action on C2C12 Cells under Hyperglycemia

Metformin is the most used biguanide drug for the treatment of type 2 diabetes mellitus. Despite being mostly known for its hepatic anti-gluconeogenic effect, it is also known to modulate microRNAs (miRNAs, miRs) associated with metabolic diseases. The latter mechanism could be relevant for better understanding metformin’s mechanisms underlying its biological effects. In the current work, we found that metformin increases miR-378a-3p expression (p < 0.002) in C2C12 myoblasts previously exposed to hyperglycemic conditions. While the inhibition of miR-378a-3p was shown to impair metformin’s effect in ATP production, PEPCK activity and the expression of Tfam. Finally, mitophagy, an autophagic process responsible for the selective degradation of mitochondria, was found to be induced by miR-378a-3p (p < 0.04). miR-378a-3p stimulated mitophagy through a process independent of sestrin-2 (SESN2), a stress-responsible protein that has been recently demonstrated to positively modulate mitophagy. Our findings provide novel insights into an alternative mechanism of action of metformin involving miR-378a-3, which can be used in the future for the development of improved therapeutic strategies against metabolic diseases.     

氢氧化钙和铝酸钠脱硫的热力学研究

采用迭代最小二乘回归法计算了一元和三元型含水硫铝酸钙的Gibbs自由能和其反应Gibbs自由能,分别为G3 CaO.Al2O3.CaSO4.12H2O=-8 894.5 kJ/mol,G3 CaO.Al2O3.3CaSO4.6H2O=-9 994.5 kJ/mol。铝酸钠溶液的脱硫反应的Gibbs自由能ΔGr1=-104.96 kJ/mol和ΔGr2=-105.54 kJ/mol,两式中ΔGr<0。从计算结果可知,有望实现以生成三元型硫酸铝钙的形式脱硫,从而使脱硫的效率提高3倍且降低AlO损失。     

全氟己酮抑制航空煤油燃烧实验及化学动力学研究

为了研究全氟己酮对航空煤油燃烧的抑制作用,将杯式燃烧器的燃烧方式由液面燃烧改为灯芯燃烧,解决了气体灭火剂灭火性能测试中较高闪点燃料难以点燃的问题。通过实验可知,随着空气中全氟己酮浓度的增加,航空煤油的燃烧经历了火焰先缓慢增高再迅速降低的过程,可见全氟己酮在不同浓度下对燃烧的作用存在由促进到抑制的转变。为深入探索这一转变的原因,基于化学动力学构建了1403个组分、7496个反应组成的全氟己酮抑制RP-3航空煤油燃烧机理并进行了验证。通过化学动力学分析可知全氟己酮在低温下对燃烧的抑制比在高温下的效果更好,全氟己酮在低浓度时温度升高导致抑制作用减弱主要是源于温度升高后,促进燃烧的反应提速幅度远大于其他反应;全氟己酮降低RP-3航煤燃烧温度的途径之一是通过热分解等吸热反应来实现的;随着全氟己酮浓度的增大,反应路径发生变化,使得H、O和OH自由基的生成量减少、消耗量大量增多,宏观上体现出全氟己酮作为燃料的促进燃烧的作用减弱、作为灭火剂抑制燃烧的作用增强。研究结果可为利用全氟己酮防控航空煤油火灾提供理论指导,为研制新型灭火剂提供参考。     

牛布鲁菌L7/L12蛋白的原核表达及纯化

目的原核表达并纯化牛布鲁菌L7/L12蛋白。方法提取牛布鲁菌S19基因组DNA,PCR法扩增L7/L12基因,经测序正确后,克隆至pET-28a(+)载体,构建重组原核表达质粒pET-L7/L12,转化大肠杆菌BL21(DE3),IPTG诱导表达。SDS-PAGE分析目的蛋白的表达形式及表达量,经亲和层析纯化后,Western blot鉴定纯化产物。结果所构建的重组原核表达质粒pET-L7/L12经酶切鉴定表明构建正确。重组蛋白以包涵体形式表达,表达量占全菌总蛋白的33%;纯化的重组蛋白纯度可达94%,可与布鲁菌小鼠免疫血清发生特异反应。结论已成功表达并纯化了牛布鲁菌L7/L12蛋白,为建立高特异性的新型布鲁菌检测方法奠定了基础。