Quantitative assessment of muscle mass and gene expression analysis in dogs with glucocorticoid-induced muscle atrophy

The present study aimed to quantitatively evaluate muscle mass and gene expression in dogs with glucocorticoid-induced muscle atrophy. Five healthy beagles received oral prednisolone for 4 weeks (1 mg/kg/day), and muscle mass was then evaluated via computed tomography. Histological and gene expression analyses were performed using biopsy samples from the biceps femoris before and after prednisolone administration. The cross-sectional area of the third lumbar paraspinal and mid-femoral muscles significantly decreased after glucocorticoid administration (from 27.5 ± 1.9 to 22.6 ± 2.0 cm² and from 55.1 ± 4.7 to 50.7 ± 4.1 cm², respectively; P<0.01). The fast- and slow-twitch muscle fibers were both atrophied (from 2,779 ± 369 to 1,581 ± 207 μm² and from 2,871 ± 211 to 1,971 ± 169 μm², respectively; P<0.05). The expression of the growth factor receptor-bound protein 10 (GRB10) significantly increased after prednisolone administration (P<0.05). Because GRB10 suppresses insulin signaling and the subsequent mammalian target of rapamycin complex 1 activity, increased expression of GRB10 may have resulted in a decrease in protein anabolism. Taken together, 1 mg/kg/day oral prednisolone for 4 weeks induced significant muscle atrophy in dogs, and GRB10 might participate in the pathology of glucocorticoid-induced muscle atrophy in canines.     

VD在骨骼外系统中的作用及其研究进展

以往普遍认为VD的主要功能是调节Ca离子来促进骨骼的生长和预防佝偻病,但最近几年的研究发现,VD不仅在骨骼疾病中起到重要作用,还与多种骨骼外疾病密切相关,包括一些心血管疾病、代谢紊乱有关的疾病、恶性肿瘤、过敏性疾病、自身免疫性疾病、生殖方面影响等。目前对VD生物学功能的研究比较全面和深入,但大部分是针对人体和小鼠的基础研究,对诸如牛、猪、犬、猫等不同动物VD的骨骼外系统生物学功能的研究较少。在查阅相关文献的基础上,通过对VD在血压、免疫调节、内分泌、子宫内膜和骨骼肌方面的生物学功能和研究进展进行总结,为不同动物VD在骨骼外系统生物学功能的研究提供一定参考。     

鱼游泳能力对体长的响应及其在鱼道设计中的应用

为了探讨鱼类体长对游泳能力的影响并为鱼道水流的设计提供参考,该研究在封闭水槽中使用"递增流速法"测试了海南省某水利枢纽鱼道目标对象的游泳能力,并用Origin软件进行了数据统计分析,得到了试验鱼感应流速、临界游泳速度和爆发游泳速度的直线回归方程和Kaplan-Meier曲线。结果表明:1)随着试验鱼体长增大,相对感应流速、临界游泳速度和爆发游泳速度(体长/s)均减小,体长和鱼类速度的相关关系可用直线方程表示,且数据经过对数变换后的直线方程拟合效果比未经过对数变换的拟合效果更好,其中R2由0.664～0.725提高至0.907～0.933。2)根据鱼道设计规范、导则及文献,结合本工程目标过鱼对象的感应流速、临界游泳速度和爆发游泳速度(m/s),建议本工程鱼道进口诱鱼流速控制在0.35～0.47 m/s,池室流速控制在0.21～0.59 m/s,竖缝流速控制在0.57～0.74 m/s,出口断面至下一个池室之间的流速控制在0.21～0.50 m/s。鱼类体长对相对游泳速度(体长/s)产生了负面影响,鱼类游泳速度及其变化规律可对鱼道水流设计值提供参考。     

利用单根肌纤维法分离和培养猪骨骼肌卫星细胞及其成肌诱导分化

建立单根肌纤维法体外培养猪骨骼肌卫星细胞的体系,了解其增殖和成肌特性。通过Ⅰ型胶原酶消化,从猪骨骼肌中分离完整的单根肌纤维并培养,用细胞免疫荧光鉴定肌纤维上的卫星细胞,随后对从单根肌纤维上游离出来的卫星细胞进行细胞免疫荧光染色,传代培养,成肌诱导分化和Western blot分析骨骼肌卫星细胞成肌特异性蛋白的表达。结果显示:分离并培养的单根肌纤维上附着有卵圆形的细胞,并随时间的推移,细胞缓慢向外迁移并增殖,卫星细胞特异性标志基因对盒转录因子(Paired protein box,Pax7)和成肌分化抗原(Myogenic Differentiation Antigen,MyoD)免疫荧光染色呈阳性,且阳性率达到90%以上。成肌诱导分化后,细胞开始汇合,并呈方向性生长,最终形成多核肌管,且成肌特异性标志基因Myogenin和myosin heavy chain(MyHC)表达呈阳性。MyoD蛋白高表达于增殖期,而Myogenin和MyHC在进入分化期才表达。该实验成功建立了猪骨骼肌单根肌纤维的体外培养方法并获得了高纯度的卫星细胞,为骨骼肌卫星细胞进行活体移植治疗相关疾病研究提供了实验材料。     

自然水体中超声波标记鱼游动轨迹精密确定算法

针对鱼类关键生境位置确定的应用需求,该文提出了一套适用于自然水体的超声波标记鱼定位算法,解决了标记鱼定位以及存在粗差观测值,即水听器记录的超声波信号接收时间存在错误情况下算法的抗干扰性。宜昌黄柏河的实测结果表明,基于现有1 ms级精度的水听器,可在自然水体中获得2.15 m精度的信号标记鱼三维游动轨迹。如因气泡、遮挡等因素对水听器观测数据引入粗差,当粗差量级在10 m以上,该方法可接近100%探测出是否存在粗差。当粗差观测值在3个以内时,该方法的探测成功率可达84.3%以上,3个以上时粗差探测成功率明显下降,5个及以上,即粗差观测值个数占观测值总数的比例大于31.25%时,基本只能探测出观测数据中存在粗差而无法有效确定粗差。该研究可为渔业增殖、鱼类栖息地保护、鱼类洄游通道等研究提供参考。     

智能气动肌肉的静态驱动特性研

研究了形状记忆合金丝(SMA)编织网的主动变形及对气动肌肉静态驱动特性的影响.建立了SMA丝内应力与静态驱动力的平衡方程.针对升降温中SMA丝的相变,分析了马氏体和奥氏体体积比变化,建立了温度SMA收缩率模型,并应用到气动肌肉的收缩率、收缩力和刚密度特性计算中.Matlab仿真得到了SMA变形曲线和气动肌肉特性变化曲面.分析结果表明SMA变形存在迟滞,SMA主动伸缩使得编织角变化范围更广,刚密度变化更突出.SMA收缩率变大,气动肌肉收缩力增强.     

cDNA cloning and complete primary structures of myosin heavy chains from brushtooth lizardfish Saurida undosquamis and wanieso lizardfish S. wanieso fast skeletal muscles

ABSTRACT:   The complete cDNA sequences encoding predominant types of myosin heavy chain (MYH) in the fast skeletal muscle were determined for brushtooth lizardfish Saurida undosquamis and wanieso lizardfish S. wanieso , which are used as materials for preparing high-quality surimi-based products. The cDNA consisted of 5973 and 5987 bp, respectively, and both encompassed an open reading frame encoding a polypeptide of 1936 amino acid residues. Brushtooth and wanieso lizardfish MYH showed the amino acid sequence identity of 92–93% to white croaker MYH, which was higher than that of 90% to walleye pollack MYH. The putative binding sites for ATP, actin, and regulatory and essential light chains in the subfragment-1 region of brushtooth lizardfish MYH exhibited a high identity with white croaker counterparts as well as the sequences of subfragment-2 and light meromyosin. In contrast, phylogenetic tree, constructed by the neighbor-joining method based on mitochondrial 16S rRNA gene, revealed that the two lizardfish species formed a cluster with walleye pollack, which was paraphyletic with white croaker. Therefore, a good reputation for lizardfish and white croaker to have a high thermal-gel forming ability seemed to be reflected by MYH rather than biological similarity as revealed by the mitochondrial 16S rRNA gene.     

Effect of xeroderma pigmentosum group D gene on proliferation of human umbilical arterial smooth muscle cells induced by Ox-LDL

AIM: To investigate the effects of xeroderma pigmentosum group D (XPD) gene on the proliferation of human umbilical arterial smooth muscle cells (HUASMCs) induced by oxidized low-density lipoprotein (Ox-LDL). METHODS: The recombinant plasmid pEGFP-N2/XPD was transfected into HUASMCs by liposome. The cells were divided into blank control group, pEGFP-N2 group, pEGFP-N2/XPD group, Ox-LDL group, Ox-LDL+pEGFP-N2 group and Ox-LDL+pEGFP-N2/XPD group. The proliferation rate of the cells was detected by MTT and EdU assays. The apoptotic rate and cell cycle distribution were analyzed by flow cytometry. The protein levels of XPD, caspase-3, Bcl-2 and Bax were determined by Western blot. RESULTS: Compared with blank control group, the expression of XPD was increased in pEGFP-N2/XPD group (P<0.05). According to the results of MTT and EdU assays, the cell proliferation in pEGFP-N2/XPD group was reduced compared with blank control group (P<0.05). Compared with Ox-LDL group, the cell proliferation in Ox-LDL+pEGFP-N2/XPD group was significantly inhibited (P<0.05). According to the results of flow cytometry, the cell proportion of S phase decreased and the G₀/G₁-phase cell proportion increased significantly in pEGFP-N2/XPD group and Ox-LDL+pEGFP-N2/XPD group compared with blank control group and Ox-LDL group, repectively (P<0.05). Compared with blank control group and Ox-LDL group, the protein level of Bcl-2 decreased and the protein levels of Bax and cleaved caspase-3 increased in pEGFP-N2/XPD group and Ox-LDL+pEGFP-N2/XPD group, respectively (P<0.05). CONCLUSION: XPD inhibits the proliferation of HUASMCs and promotes their apoptosis, and reduces the promoting effect of Ox-LDL on the proliferation of HUVSMCs. XPD may be the target for treatment of atherosclerosis.     

Model construction of rat coronary artery smooth muscle cell endoplasmic reticulum stress induced by thapsigargin

AIM: To investigate the primary culture method for coronary artery smooth muscle cells (CASMCs), and to establish the endoplasmic reticulum stress (ERS) model in CASMCs of SD rats. METHODS: CASMCs were cultured by tissue explant method. The morphological characteristics were observed under optical microscope. The marker proteins of CASMCs, including α-SMA and SM-MHC, were identified by immunofluorescence technique. The protein expression levels of BiP and CHOP, the marker molecules of ERS, were determined by Western blot. RESULTS: The spindle-shaped CASMCs climbed out from the edge of coronary artery tissues after 6 d, and formed the typical "hill and valley" growth pattern of CASMCs at 9~10 d. The result of immunofluorescence technique showed that α-SMA and SM-MHC were positively expressed. The results of Western blot showed that the protein expression of BiP and CHOP in TG (1 and 2 μmol/L) treatment groups was increased compared with control group. Compared with control group, the protein expression of BiP and CHOP was significantly increased after 1 μmol/L TG treatment for 24 and 48 h. CONCLUSION: CASMCs can be successfully cultured by tissue explant method. ERS model of CASMCs was established by 1 μmol/L TG treatment for 24 h.     

Farrerol inhibits nicotine-induced proliferation of pulmonary vascular smooth muscle cells by enhancing Kv1.5 and Kv2.1 expression

AIM:To study the effect of farrerol (Far) on nicotine-induced proliferation of rat pulmonary smooth muscle cells (PASMCs), and further to explore its relationship with voltage-dependent potassium channels (Kv) 1.5 and Kv2.1. METHODS:Firstly, the effect of nicotine on the proliferation of PASMCs was detected by cell counting method, and the optimal concentration of nicotine was selected. Primary cultured PASMCs were randomly divided into 5 groups:normal control group, nicotine (1 μmol/L)group, nicotine (1 μmol/L) + Far (10^-6 mol/L, 10^-5 mol/L and 10^-4 mol/L) Far group. The activity of caspase-3 was measured by apoptosis kit, the cell viability was measured by CCK-8 assay, the apoptotic rate was analyzed by flow cytometry. The expression of Kv1.5 and Kv2.1, and apoptosis-related factors Bcl-2 and Bax at mRNA and protein levels was determined by RT-qPCR and Western blot respectively. RESULTS:Nicotine at 1 μmol/L increased the number of PASMCs to the maximum extent (P<0.01). Nicotine at 1 μmol/L significantly reduced the caspase-3 activity and enhanced the cell viability of the PASMCs (P<0.01). Farrerol at 10^-6~10^-4 mol/L eliminated the effect of PASMCs induced by nicotine in a concentration dependent manner. Compared with control group, nicotine at 1 μmol/L significantly increased the proliferation and inhibited the apoptotic rate of rat PASMCs (P<0.01). The apoptotic rate of PASMCs in farrerol intervention group was significantly higher than that in nicotine group (P<0.01). Nicotine at 1 μmol/L significantly inhibited the expression of Kv1.5, Kv2.1 and Bax but increased the expression of Bcl-2 in PASMCs (P<0.01). Farrerol at 10^-5 mol/L obviously inhibited the effect of PASMCs induced by nicotine. CONCLUSION:Farrerol eliminates nicotine-induced inhibition of caspase-3 and Bax, and enhancement of Bcl-2 in PASMCs by enhancing Kv1.5 and Kv2.1 expression.