The Expression of Long Noncoding RNA (H19) in Mouse Skeletal Muscle Development

This experiment was intend to study the changes of long non-coding RNA (H19) expression levels in skeletal muscle development and regeneration,and lay the foundation of its mechanism reach in skeletal muscle development.C2C12 cell line and ICR mice were used as experimental material,bioinformatics assay was used to exploit its non-coding character and low conservatism in different species,and the expressions of H19 in C2C12 cell differentiation,skeletal muscle development and the phase of muscle regeneration were detected by quantitative Real-time PCR (qRT-PCR).The results showed that,H19 expression levels in postnatal mouse skeletal muscle decreased with increasing age;during C2C12 cell differentiation,H19 mRNA increased gradually,and then maintained a high level;The expression of H19 was maintained at a high level through days 4 to 6 after injury.In consideration of its express character in C2C12 cell differentiation and skeletal muscle damage repair model,H19 may play an important role in promoting myogenesis and skeletal muscle regeneration.     

Analysis of Regular Nutritional Components and Amino Acid Contents of Mongolia Bullock Muscle Grazed in Kubuqi Desert

In order to evaluate the Mongolia muscle characteristics and nutritional value, six Mongolia bullocks aged 4 years grazed in Kubuqi desert were chosen to analyze the regular nutritional components and amino acid contents of Mongolia muscle.The results showed that the contents of water, crude protein (CP), crude fat (EE), crude ash (ash), calcium and phosphorus of Mongolia muscle (fresh) were 68.21%, 22.60%, 6.49%, 2.69%, 0.58% and 0.55%, respectively.A total amino acid (TAA) content of beef was 24.297 g/100 g, essential amino acid content was 10.265 g/100 g, the proportion of essential amino acid justified and exceeded the FAO/WHO standard.The ratio of essential amino acid and total amino acid contents (EAA/TAA) was 42.25%, the ratio of essential amino acid and the non-essential amino acid (EAA/NEAA) was 73.15%.In functional amino acid, the content of palatable taste amino acid and sweet flavor amino acid in total amino acid was 46.891%.In addition to valine, essential amino acid contents of Mongolia muscle were superior to ideal protein of FAO/WHO standard, the percentage of valine reached 83.972% in ideal protein.These results indicated that Mongolia muscle belonged to the comprehensive nutrition and high quality protein food on the evaluation of the regular nutrient content and the amino acid content of beef.     

湘中丘陵区马尾松林粗木质残体研究

以湘中丘陵区马尾松林为研究对象,分析粗木质残体(CWD)的储量特征、物种组成、径级结构以及腐烂等级,研究湘中丘陵区马尾松林CWD的基本特征。结果表明:湘中丘陵区马尾松林CWD的平均储量为1.72 t/hm2,马尾松和杉木是针叶CWD的主要组成树种,樟树、南酸枣、枫香是阔叶CWD的主要组成树种。马尾松-阔叶树种混交林的CWD总储量显著大于马尾松-杉木混交林的,两者的CWD总储量与其对应森林总生物量的比例差异明显,且均表现为随森林生物量增长而增加的趋势。CWD的个体数量随着径级的增加而减少,呈反J型下降趋势,且不同存在形式CWD的数量比例大致表现为倒木枯立木大枝根桩。CWD具备完整的腐烂等级(Ⅰ~Ⅴ级),主要集中在Ⅰ、Ⅱ腐烂等级,且随着径级的增加,各腐烂等级CWD的数量比例呈现逐渐减少的趋势。     

Zinc supplementation strategies in feedlot heifers receiving an extended-release implant or an aggressive re-implant program

Two hundred eight Angus-crossbred heifers (291 ± 23 kg) from four sources were used in a randomized complete block design. The objective of the study was to determine the effects of implant strategy and Zn supplementation on performance, carcass characteristics, muscle fiber diameter, and mineral status of heifers. Heifers were assigned to a 2 × 2 factorial study for 168 d, and factors included Zn and implant (IMP). Heifers were supplemented Zn (mg/kg dry matter [DM]; ZnSO₄) at national (30; NRC) or industry (100; IND) recommendations. Implant strategies (Merck Animal Health, Madison, NJ) included extended-release Revalor-XH on day 0 (REV-XH; 20 mg estradiol + 200 mg trenbolone acetate) containing four uncoated pellets and six coated pellets or the uncoated implant Revalor-200 on day 0 and again on day 91 (REV-200; 20 mg estradiol + 200 mg trenbolone acetate). Heifers were blocked by weight within source to pens of five or six heifers per pen (nine pens per treatment). A corn silage-based diet was fed during the growing period (days 0–55) followed by transition to a corn-based finishing diet. Weights were taken consecutively on days −1/0, 55/56, and 167/168. Liver and muscle from the longissimus thoracis were collected from one heifer per pen on days −5, 14, 105, and 164. Data were analyzed via Mixed Procedure of SAS. Average daily gain (ADG) and liver mineral used Period as the repeated effect. Corresponding to periods of high hormone payout from each implant, days 0–28 and 91–120 ADG were greatest for REV-200, whereas REV-XH numerically peaked during days 56–91 (IMP × Period; P = 0.02). Day 91 IND body weight tended to be heavier (P = 0.06) and day 120 body weight was heavier (P = 0.05) than NRC heifers. No effect of Zn or IMP on final body weight was observed (P ≥ 0.21). Muscle fiber cross-sectional diameter on day 164 was greater (P = 0.05) in IND than NRC. Liver Mn concentrations decreased by day 14 regardless of implant, though days 105 and 164 concentrations were lesser for REV-200 than REV-XH (IMP × Period; P = 0.02). No effects of Zn, IMP, or the interaction were observed for carcass-adjusted gain to feed, days 0–168 DM intake, hot carcass weight, or ribeye area (P ≥ 0.11). The nominal differences in performance between implant strategies suggest that extended-release implants may be an effective implant strategy to replace re-implant programs in heifers, whereas the improved performance of heifers fed IND vs. NRC during times of peak hormone payout suggests a role for Zn in periods of rapid growth.     

IL-15过表达对猪骨骼肌细胞成肌分化的影响

【目的】研究肌肉因子IL-15（interleukin 15）对猪骨骼肌成肌细胞增殖与凋亡的影响,为进一步研究IL-15在动物肌肉品质调控和骨骼肌疾病治疗提供依据。【方法】构建IL-15过表达慢病毒载体GV-492-IL-15,体外无菌分离培养猪骨骼肌卫星细胞,诱导分化,并通过免疫荧光染色进行成肌细胞验证。将成肌细胞转染IL-15过表达重组慢病毒载体,试验分别设置空白对照组（Control）、转染阴性对照病毒组（IL-15-）和转染GV-492-IL-15（IL-15+）慢病毒试验组（n=3）。培养72 h后,收集细胞和培养上清液。分别采用实时定量PCR（qRT-PCR）和Western Blot技术分析目的基因和蛋白的表达情况,采用ELISA试剂盒分析培养液中IL-15含量,采用CCK-8试剂盒分析成肌细胞活力,采用流式细胞术分析细胞周期和细胞凋亡,采用Western Blot技术检测与细胞凋亡密切相关的caspase-3蛋白表达水平的变化。【结果】（1）经鉴定后的质粒转染293T细胞,细胞内可观察到明显的绿色荧光,经Western Blot检测,可以观察到20 kD附近处有特征条带;（2）分离培养的猪骨骼肌卫星细胞呈梭形或纺锤形,诱导后可分化为呈管状的成肌细胞。将分化后的成肌细胞,进行α-SMA单克隆抗体免疫荧光染色,视野中90%的细胞呈阳性反应,胞浆染成红色,表明细胞为骨骼肌成肌细胞。（3）转染GV-492-IL-15慢病毒后,与对照细胞组相比,成肌细胞内IL-15 mRNA和蛋白相对表达量均极显著升高（P<0.001）,但培养液中IL-15蛋白水平变化不大（P>0.05）。CCK-8结果显示,过表达IL-15可增强细胞的增殖能力（P<0.05）。与对照组相比,转染GV-492-IL-15慢病毒的细胞早期凋亡率差异不显著（P>0.05）,但细胞晚期凋亡率显著下降（P<0.05）。与对照组相比,转染慢病毒组细胞中caspase-3蛋白有下降的趋势,但差异不显著（P>0.05）。此外,转染IL-15过表达慢病毒可使G1期细胞比例显著下降,S期和G2/M期细胞比例显著升高（P<0.05）。【结论】在正常生理条件下,IL-15是定位在细胞内并发挥作用的,IL-15过表达对猪骨骼肌成肌细胞早期凋亡没有显著影响,但可以抑制其晚期凋亡,并促进细胞增殖。这一研究将为IL-15正向调控猪骨骼肌肌肉品质和治疗相关肌肉疾病提供技术和理论依据。     

荨麻皮刺水提液对小鼠离体肠平滑肌运动的影响

【目的】研究荨麻促进消化道平滑肌运动的植株主要部位,分析其可能的受体机制。【方法】用生物信号处理系统监测麻叶荨麻皮刺与内茎(不含皮刺)水提液对小鼠离体小肠平滑肌的影响。【结果】2倍浓度递增(0.1%~3.2%)的皮刺剂量依赖性增加小肠收缩幅度、收缩张力和收缩峰值(P<0.05),但对收缩频率无影响(P>0.05);而内茎浓度达0.4%后使小肠收缩频率减慢(P<0.05)。0.8%皮刺一次性用药后小肠收缩幅度、收缩张力、收缩峰值均显著升高(P<0.05),而同剂量内茎对小肠收缩无影响(P>0.05)。M-受体阻断剂ATr与0.8%皮刺联合用药可使收缩幅度显著减弱(P<0.05)。【结论】荨麻植株的皮刺对消化道平滑肌具有促进作用,且可能部分作用是通过激活M-受体而实现。     

零换水条件下复合益生菌对罗非鱼生长性能、肌肉品质及养殖水体环境的影响

【目的】研究零换水条件下复合益生菌对罗非鱼生长性能、肌肉品质及养殖水体环境的影响,为渔 业节能减排与生态高效健康养殖提供参考。【方法】以罗非鱼为研究对象,在零换水条件下添加复合益生菌（试 验处理）开展罗非鱼养殖试验,以正常换水的养殖模式作为对照,进行罗非鱼生长性能、肌肉品质及主要水质指 标、细菌群落结构的对比。【结果】在零换水条件下,试验处理罗非鱼成活率为 83.3%,显著高于对照;饵料系 数 2.3,显著低于对照;肌肉品质方面,试验处理粗蛋白含 22.17%,显著高于对照。在整个养殖周期,试验处理 的主要水质指标维持相对稳定,水温为 26.5~30.5℃,pH 为 7.0~8.0,而亚硝酸盐为 0.039 mg/L,显著低于对照; 两组优势菌群相似,但试验处理菌群多样性指数高于对照。【结论】在零换水条件下,养殖水体中添加复合益生 菌提高了罗非鱼的成活率,降低了饵料系数,并在一定程度上改善了肌肉品质,并对养殖水环境具有改善作用, 菌群多样性指数提高,在整个养殖生产周期内对常规养殖模式依靠频繁换水释放环境压力的替代作用良好。     

苏尼特羊不同生长时期肌肉组织lncRNA的差异表达分析

为鉴定和分析苏尼特羊肌肉不同发育时期lncRNA的差异表达,利用RNA-seq技术对120d苏尼特羊胎儿和5月龄羔羊肌肉组织进行了lncRNA比较分析,筛选出差异表达的lncRNA及其靶基因,并对预测的mRNA进行了GO和KEGG分析,同时对随机筛选的差异表达lncRNA进行qRT-PCR验证。结果显示,120d和5月龄羔羊肌肉之间差异表达的lncRNA数共为626个,共靶向1 273个mRNA编码基因。GO和KEGG分析表明,这些差异基因主要涉及代谢过程的调控,生物合成,基因表达,蛋白结合以及mTOR信号通路、FoxO信号通路、胰岛素信号、肌动蛋白细胞骨架的调节、Wnt信号通路和间隙连接等信号通路。qRT-PCR验证证明,随机选取的8个lncRNA在肌肉组织中的表达量与RNA-seq结果一致。综上,本研究利用RNA-Seq技术分析了苏尼特羊120d胎儿和5月龄羔羊的差异表达lncRNA及其靶基因,发现其参与了不同生长时期苏尼特羊的肌肉发育和生长过程,为从lncRNA的角度更好地理解苏尼特羊肌肉生长发育的遗传调控机制提供了理论基础,也为绵羊分子辅助育种提供参考。     

灌喂必需氨基酸模式溶液对草鱼全鱼_省略_肌肉_肝胰脏蛋白质合成代谢的影响

采用大剂量食道灌喂DL－[4-^3H]-Phe，研究不同EAA模式下，20－30g体重草鱼种的全鱼和肌肉，肝胰脏的蛋白质合成代谢，结果表明：（1）分别缺乏Lys,Met,Trp Arg的4种极端不平衡EAA模式，同草鱼肌肉EAA模式即相对平衡的基础模式相比，肌肉，肝胰脏及全鱼的蛋白质合成速率（FSR）均显著降低，其降低程度高低依次为Met,Lys,Arg,Trp;(2)肝胰脏，肌肉人的蛋白质合成代谢与整体蛋白质合成代谢对EAA模式的改变产生相同的变化趋势，并且三者蛋白质合成代谢速率为肝胰脏＞全鱼＞肌肉；（3）EAA模式的Lys,Met,Trp,Arg4因素中，对肌肉，肝胰脏蛋白质合成速率影响最大的是Met,其余依次为Lys,Arg,Trp。     

散鳞镜鲤、团头鲂及其杂交F1肌肉营养成分的比较

采用生化分析方法对散鳞镜鲤、团头鲂及其杂交F1肌肉的营养成分进行了测定并比较。结果表明，交杂种F1肌肉中的水分含量为77．89％、灰分含量为5．52％，均低于亲本；脂肪含量为4．77％，高于团头鲂，低于散鳞镜鲤；F1的氨基酸含量为77．89％，低于双亲，但大多成分含量与散鳞镜鲤相当。在总体水平上，杂交种品质得到初步的改良。