生肌调节因子在绵羊不同胎儿期心肌和骨骼肌中的表达

本试验旨在探究生肌调节因子（myogenic regulatory factors，MRFs）在绵羊不同胎儿发育时期心肌和骨骼肌组织中的表达情况。选取妊娠60（E60）、70（E70）、80（E80）和90 d（E90）4个不同时期的滩羊胎儿作为研究对象，采集骨骼肌和心肌组织，通过HE和油红染色观察心肌和骨骼肌在胎儿发育过程中发生的组织形态学变化；通过实时荧光定量PCR研究绵羊胎儿发育过程中MRFs在心肌和骨骼肌组织中的表达规律以及同一胎儿期MRFs在心肌和骨骼肌中的表达差异。HE染色结果显示，心肌组织和骨骼肌组织的微观结构差异明显，心肌肌纤维密度大，纵横交错成网状，和骨骼肌相比发育的更加成熟；骨骼肌组织肌纤维密度小，间隙大，E60~E90肌纤维数量逐渐增加，肌纤维束结构清晰分明。油红染色结果显示，心肌和骨骼肌组织在4个时期均未出现被染成红色的脂滴，表明E60~E90两个组织均没有分化出脂肪。实时荧光定量PCR结果显示，E60~E90胎儿心肌组织中MYOG、Myf5和Myf6基因表达量持续降低，但是没有MYOD基因表达；E60~E90骨骼肌组织中MYOD、MYOG和Myf6基因表达量持续上升，其中MYOG基因表达水平最高，Myf5基因的表达发生周期性波动。以上结果表明，E60~E90胎儿骨骼肌正处于快速发育阶段，而心肌组织此时已大致成型。因此，骨骼肌组织中MRFs的表达量远远高于心肌组织，且MYOG基因对于维持骨骼肌的快速发育发挥了重要作用。     

甜菜碱对瘦素缺陷小鼠肌肉组织中脂质沉积的影响初探

为探究甜菜碱对瘦素缺陷（ob/ob）小鼠肌肉组织中脂质沉积的影响和潜在机制，本研究将12只同批次6周龄雄性、健康、体重差异不超过1 g的ob/ob小鼠随机均分为两组，分别作为试验组和对照组。其中对照组小鼠饮用纯净水，试验组小鼠饮用的纯净水中含1%甜菜碱。1个月后脱颈处死，采集肌肉样本，利用油红O染色、甘油三酯含量测定检测ob/ob小鼠肌肉组织中脂质沉积情况；利用气象色谱-质谱联用系统分析肌肉组织中脂肪酸组成及含量；利用qRT-PCR方法检测肌肉组织中基因表达的变化。结果显示：与对照组相比，1）补饲甜菜碱后ob/ob小鼠的体重增加量、肌肉组织中脂质沉积、禁饲血糖水平显著降低（P<0.05）；2）补饲甜菜碱后，ob/ob小鼠肌肉中脂肪甘油三酯脂肪酶（adipose triacylglyceride lipase，ATGL）、激素敏感性甘油三酯脂肪酶（hormone-sensitive lipase，HSL）以及葡萄糖转运蛋白4（glucose transpoter 4，GLUT4）基因的表达量极显著升高（P<0.01）；3）补饲甜菜碱后，ob/ob小鼠肌肉中饱和脂肪酸（saturated fatty acids，SFA）和多不饱和脂肪酸（polyunsaturated fatty acids，PUFA）含量显著升高（P<0.05）；4）补饲甜菜碱后，ob/ob小鼠肌肉中脂肪酸合成基因：脂肪酸合成酶（fatty acid synthetase，FAS）表达量显著降低（P<0.05），乙酰辅酶A合成酶（acetyl-CoA synthetase，ACS）表达量无显著性变化（P>0.05）；脂肪酸氧化基因：过氧化物酶体增殖物激活受体α（peroxisome proliferator activated receptor alpha，PPARα）、酰基辅酶A氧化酶（acyl-CoA oxidase2，ACOX2）、长链酰基辅酶A脱氢酶（long chain acyl-CoA dehydrogenase，ACADL）、脂肪酸转位酶（fatty acid transposase， CD36）表达量极显著升高（P<0.01）。结果表明，ob/ob小鼠补饲甜菜碱后，肌肉组织脂肪酸合成基因表达降低、脂肪酸氧化基因表达升高，同时肌肉组织脂肪酸含量发生改变，最终导致肌肉组织脂质减少。     

外源注射miR-499-5p对肉鸡腓肠肌转录组的影响

旨在探讨外源注射miR-499-5p对肉鸡腓肠肌肌纤维直径及基因表达的影响，完善miR-499-5p对肉鸡骨骼肌发育的分子调控机制。本研究选用14日龄健康大恒肉鸡公鸡16只，随机分为2组，每组8个重复，均正常饲喂日粮，分别在腓肠肌部位肌肉注射agomiR-NC（对照组，5 nmol）和agomiR-miR-499-5p（注射组，5 nmol），为保证试验效果，每3 d注射1次，共注射6次。注射完成后将16只肉鸡屠宰，采集腓肠肌组织进行肌纤维直径测定和转录组学（RNA-seq）分析。结果显示，注射miR-499-5p可导致肉鸡腓肠肌肌纤维直径显著减小（P<0.05）。转录组测序结果显示，注射组和对照组分别获得308 263 164和316 696 152条clean reads，与鸡参考基因组的比对率分别为90.29%和89.57%。与对照组相比，注射组共有差异表达基因18个，其中上调基因12个，下调基因6个（P<0.05）。其中差异表达最大的两个基因分别是原癌基因FOS（fos proto oncogene，FOS）和早期生长应答蛋白基因（EGR1）。GO富集分析发现，差异表达基因显著富集于跨膜受体蛋白丝氨酸/苏氨酸激酶信号通路、转录调控、脂质合成和骨骼肌再生等过程。KEGG富集分析结果显示，差异表达基因显著富集于丝裂原活化蛋白激酶（MAPK）信号通路和Wnt信号通路。利用生物信息学软件预测到miR-499-5p的靶基因共78个，GO富集分析发现，靶基因也显著富集于丝氨酸/苏氨酸激酶活性的负调控和转录因子活性等过程。本研究表明，注射miR-499-5p可改变肉鸡腓肠肌肌纤维直径从而影响肌纤维特性，差异表达基因中的FOS、EGR1、CYR61和WNT7A基因可能在miR-499-5p的作用下通过上述通路参与了脂肪沉积和肌肉发育的调控。     

Correlation between skeletal muscle fiber type and responses of a taste sensing system in various beef samples

The difference of muscle fiber type composition affects several parameters related to meat quality; however, the relationship between muscle fiber types and meat taste is unclear. To elucidate this relationship, we determined the taste of various beef samples using a taste sensor (INSENT SA402B) and analyzed its correlation with different muscle fiber type composition. We used 22 kinds of beef samples and measured nine tastes, including the relative and change of membrane potential caused by adsorption (CPA) values, using six sensors (GL1, CT0, CA0, AAE, C00, and AE1). The taste sensor analysis indicated positive value outputs for the relative C00, AAE, and GL1 values as well as for the CPA value of AAE, which corresponded to bitterness, umami, sweetness, and richness, respectively. We found significant positive correlations of the myosin heavy chain 1 (MyHC1) composition with umami taste, and with richness. This result suggests that high levels of slow MyHC1 can induce strong umami taste and richness in beef. We expect that our results will contribute to the elucidation of the relationship between muscle fiber types and meat palatability.     

Effects of growth hormone treatment on the expression of somatotropic axis genes in the skeletal muscle of lactating Holstein cows

This study focused on the expression of somatotropic axis genes in the skeletal muscle of dairy cattle. A slow-release recombinant bovine growth hormone (GH) (rbGH) formulation was administered to 5 cows, and saline solution (control) was administered to another 5 cows every 2 wk for a total of 10 wk, starting from the peak of lactation. Tissue and blood samples were collected on days 2 and 14 after each rbGH injection. As target genes insulin-like growth factor (IGF)-1, IGF-2, IGFBPs (1, 2, 3, 4, 5, 6), acute labile subunit (ALS), IGF-1 receptor (IGF-1R), GH receptor (GHR), and the known GHR 5′-UTR variants were selected as target genes, and their relative expression was measured using real-time polymerase chain reaction. In GH-treated cows, an increase in expression was observed for GHR 5′-UTR variant 1I on day 14 (P < 0.05), whereas a significant down-regulation of GHR (P < 0.05) was found after comparing values of treated cows between day 2 and day 14. However, only IGF binding proteins (BP)-5 was found to be appreciably up-regulated in GH-treated cows (P < 0.001), which may indicate the importance of this gene in the overall molecular response to GH administration. Our study indicated that GH treatment did not affect the expression of most somatotropic axis genes, despite the marked increase in GH and IGF-1 in blood (P < 0.001). Nor did it have a large impact on the proportion of GHR 5′-UTR variants in the skeletal muscle of lactating cows. Finally, although we observed a significant variation in the expression of some genes, it would appear that the differences between GH-treated cows and controls were not great enough to be considered as reliable indirect indicators of GH treatment in dairy cattle.     

Thy-1 Expressing Mesenchymal Cells in Rat Nephrogenesis in Correlation with Cells Immunoreactive for α-Smooth Muscle Actin and Vimentin

Thy-1 expression may influence myofibroblast development. Through the epithelial-mesenchymal transition (EMT), injured renal epithelial cells undergo regression to the metanephric mesenchymal phenotype and then acquire a myofibroblastic nature (expressing α-smooth muscle actin; α-SMA). Because the metanephric blastema differentiates into mesenchymal and renal epithelial cells, we investigated Thy-1 immunoexpression during nephrogenesis in F344 rats in correlation with vimentin and α-SMA expressions. Kidney samples were obtained from fetuses on gestation days 18 and 21, neonates on days 1-18 and adults at 6 weeks of age. Mesangial cells in S-shaped bodies and immature and mature glomeruli continuously expressed both Thy-1 and α-SMA during early nephrogenesis (fetuses and neonates on days 1-9). During early nephrogenesis, loosely-arranged blastemal cell-derived mesenchymal cells in the cortex and medulla also exhibited Thy-1 and α-SMA, although the α-SMA expression was weaker than that of Thy-1. Vimentin expression coincided with that of Thy-1. These findings indicate that the derivation of α-SMA-expressing myofibroblastic cells may be related to mesangial or blastemal cells expressing both Thy-1 and α-SMA. Interestingly, there was a difference in Thy-1 expression between cortical and medullary tubulointerstitial cells from late nephrogenesis (neonates on days 12-18) and those from adults in that the cortical cells reacted faintly or negatively to Thy-1, whereas the medullary cells reacted strongly to Thy-1; additionally, bundle-arranged mesenchymal cells that were only observed in the neonates on days 1-12 reacted strongly to α-SMA, but faintly to Thy-1. Blastemal cell-derived mesenchymal cells seem to alter the immunoexpressions of Thy-1 and α-SMA, depending on the conditions which they develop. Thy-1 immunoexpression would be useful for investigation of reverse embryogenesis, which might occur in fibrotic kidneys.     

品种、体重和营养对猪背最长肌肌纤维组织学特性的影响

选用荣昌(RC)猪和杜×长×大(DLY)杂交猪为试验对象,研究猪背最长肌肌纤维直径和密度的发育性变化及品种与营养影响特点。结果表明:2个品种猪的背最长肌肌纤维直径和密度的发育规律较为类似,肌纤维直径随体重增加呈线性增长,而肌纤维密度则随体重增加呈乘幂方程式降低。相关分析表明,肌纤维密度与直径均存在极显著负相关(P≤0.003)。背最长肌肌纤维直径和密度在20～50kg阶段品种间差异不显著(P≥0.099),但在80kg,RC猪的肌纤维直径显著低于DLY猪(P=0.0043),而RC猪的肌纤维密度则显著高于DLY猪(P=0.0049)。饲粮营养水平对2个品种猪背最长肌的肌纤维直径和密度均无显著影响。以上结果提示,2个品种猪的背最长肌肌纤维的生长规律较为类似,但不同品种的肌纤维直径存在差异,主要表现在80kg,RC猪显著低于DLY猪,可能与其优良肉质有关。     

Subcutaneous embryonal rhabdomyosarcoma in a dog: cytologic, immunocytochemical, histologic, and ultrastructural features

Abstract: A subcutaneous mass on the left antebrachium of an 11‐year‐old intact female English Pointer dog was evaluated presurgically by cytologic examination and immunocytochemical staining. The sample consisted of discrete, variably sized, markedly pleomorphic neoplastic cells that expressed vimentin with diffuse cytoplasmic staining, desmin with focal paranuclear staining, and myoglobin with diffuse cytoplasmic staining, consistent with a diagnosis of rhabdomyosarcoma. Lymphocytic and histiocytic markers were negative. Aspirates of the enlarged ipsilateral prescapular lymph node were positive for metastatic disease. Surgical excision of the tumor and lymph node were followed by histologic and electron microscopic examination. Histomorphologic appearance of neoplastic cells from the mass and the lymph node paralleled cytologic findings; the histologic diagnosis was round cell variant of embryonal rhabdomyosarcoma. By ultrastructural evaluation, cells contained numerous mitochondria and masses of cytoplasmic tangled myofilaments, features typical of rhabdomyoblasts. The dog received doxorubicin (30 mg/m²) every 3 weeks for 5 treatments. Local recurrence developed 6 months after resection but was not treated. Despite a guarded prognosis and untreated local recurrence, the dog was still alive 18 months after surgery. Cytologic evaluation and immunocytochemical staining were pivotal for the presurgical diagnosis of rhabdomyosarcoma.     

Effects of Soybean Isoflavones on In vitro Antioxidative Capacity of Satellite Cells of Porcine Skeletal Muscles

A synthetic isoflavone (ISO-S) or genistein was added in culture medium at different concentrations (0,10,20,30,40,and 80 pmol L-1) to investigate the effects of soybean isoflavones on antioxidative capacity of porcine skeletal muscle satellite cells.After 48 h incubation,the suspension was cryopreserved for the determination of superoxide dismutase (SOD),catalase (CAT),glutathione peroxidase (GSH-Px) activities,and malondialdehyde (MDA) content.The mRNA levels of SOD,CAT,and GSH-Px gene in cells were detected with Taqman fluorescent probe method.The results showed that the content of MDA and the activities and the mRNA levels of SOD of porcine skeletal muscle satellite cells were influenced by supplemented soybean isoflavone (P<0.05) when adding 10-80 gmol L-1 ISO-S or genistein in the medium.The MDA contents,SOD and CAT activities and their mRNA expression levels of porcine skeletal muscle cells responded quadratically P(<0.05) as the level of ISO-S or genistein increased.Pre-incubation of porcine skeletal muscle satellite cells with ISO-Sor genistein at 10-40 pmol L-t elevated the activities and the mRNA expression levels of SOD and CAT in cells concurrently and decreased the cellular content of MDA (P<0.05).The results indicated that pre-incubation of ISO-S or genistein at 10-40 pmol L-1 could improve the antioxidative capacity of porcine skeletal muscle satellite cells.     

广东池养梭鱼肌肉营养成分的分析及评价

对广东池养梭鱼肌肉中一般成分(水分、粗蛋白、粗脂肪、灰分)、氨基酸、脂肪酸、营养元素进行测定,并进行了较为全面的分析和比较。结果表明,其肌肉鲜样中水分含量为76.9%,粗蛋白含量为20.2%,粗脂肪含量为1.2%,灰分含量为1.13%。肌肉中18种氨基酸的总量为湿重的19.43%。其中,8种必需氨基酸的总量占湿重的8.77%,占氨基酸总量的45.14%,必需氨基酸与非必需氨基酸的比例为97.23%,符合FAO/WHO的标准。梭鱼的第一、二限制性氨基酸分别为缬氨酸和蛋氨酸+胱氨酸,必需氨基酸指数(EAAI)为61.56。梭鱼肌肉中含有22种脂肪酸,不饱和脂肪酸含量丰富,与饱和脂肪酸的比例约为3∶1;EPA+DHA的含量占干重的5.92%,高于其他一些经济鱼类;梭鱼微量元素的比例合理。