Effect of Estradiol-17β on protein synthesis and degradation rates in fused bovine satellite cell cultures

Although androgenic and estrogenic steroids are widely used to enhance muscle growth and increase feed efficiency in feedlot cattle, their mechanism of action is not well understood. Further, in vivo studies indicate that estradiol (E2) affects muscle protein synthesis and/or degradation, but in vitro results are inconsistent. We have examined the effects of E2 treatment on protein synthesis and degradation rates in fused bovine satellite cell (BSC) cultures. Additionally, to learn more about the mechanisms involved in E2-enhanced muscle growth, we have examined the effects of compounds that interfere with binding of E2 or insulin-like growth factor (IGF)-1 to their respective receptors on E2-induced alterations in protein synthesis and degradation rates in BSC cultures. Treatment of fused BSC cultures with E2 results in a concentration-dependent increase (P < 0.05) in protein synthesis rate and a decrease (P < 0.05) in protein degradation rate. The pure estrogen antagonist ICI 182 780 suppresses (P < 0.05) E2-induced alterations in protein synthesis and degradation in fused BSC cultures. The G-protein coupled receptor (GPR)-30 agonist G1 does not affect either synthesis or degradation rate, which establishes that GPR30 does not play a role in E2-induced alterations in protein synthesis or degradation. JB1, a competitive inhibitor of IGF-1 binding to the Type 1 insulin-like growth factor receptor (IGFR-1), suppresses (P < 0.05) E2-induced alterations in protein synthesis and degradation. In summary, our data show that E2 treatment directly alters both protein synthesis and degradation rates in fused BSC cultures via mechanisms involving both the classical estrogen receptor (ER) and IGFR-1.     

Multifocal cutaneous histiocytic sarcoma in a young dog and review of histiocytic cell immunophenotyping

A 9‐month‐old male Great Dane had progressive generalized nodular dermatopathy for several months. There were > 100 raised, alopecic, firm, painful nodules throughout the skin. Aspirates from several lesions yielded moderate numbers of irregularly round or polygonal to spindle‐shaped cells with mild to moderate anisocytosis and few inflammatory cells, and the cytologic interpretation was proliferation of mesenchymal or histiocytic cells. On histopathologic examination, nodules were composed of densely packed sheets of round to spindle‐shaped cells with mild anisokaryosis and low mitotic activity. Multifocal histiocytic sarcoma with a spindle‐cell pattern was diagnosed based on morphologic features and intense expression of CD18. Additional immunophenotypic analysis on frozen sections of tissue confirmed the diagnosis of histiocytic sarcoma; expression of CD18, CD45, CD1a, CD11b, and CD11c, limited expression of Thy‐1 (CD90) and CD80, and lack of expression of CD4, CD11d, and CD86 indicated that the cells were likely interstitial dendritic cells; a review of reactive and neoplastic dendritic cells is provided. Based on staging, internal organs were not affected. Sequential treatment with lomustine and doxorubicin failed to prevent progression of the cutaneous lesions, and the dog died 3 months after initial diagnosis. At necropsy, a focus of neoplastic cells was present in one lymph node, but except for skin other organs were not involved. The clinical presentation of histiocytic sarcoma may be unusual, and neoplastic cells may lack overt features of malignancy on cytologic and histopathologic examination. In some instances, immunophenotyping is required to differentiate histiocytic sarcoma from other histiocytic disorders.     

斯氏艾美耳球虫ITS1-5.8S rRNA-ITS2序列的克隆及PCR检测方法的建立

通过对多种鸡球虫和松鼠球虫18SrRNA和28SrRNA进行序列比对分析,在18SrRNA 3′端和28SrRNA 5′端保守区设计艾美耳属通用引物,以斯氏艾美耳球虫洛阳分离株LY卵囊基因组DNA为模板首次成功克隆到斯氏艾美耳球虫完整的ITS1-5.8SrRNA-ITS2序列,其大小为1 178bp,其中ITS1序列长度为423bp,5.8SrRNA为155bp,ITS2为600bp,斯氏艾美耳球虫LY株ITS1/2序列高度变异,与鸡球虫、啮齿动物球虫的序列相似性低于60%。然后在斯氏艾美耳球虫ITS1/2序列超变区设计种特异引物,建立了灵敏、特异的PCR检测方法。本研究结果将为兔球虫强致病种的临床诊断和揭示兔球虫种群遗传特征提供有效的分子工具。     

Radiosensitivity and capacity for radiation-induced sublethal damage repair of canine transitional cell carcinoma (TCC) cell lines

Understanding the inherent radiosensitivity and repair capacity of canine transitional cell carcinoma (TCC) can aid in optimizing radiation protocols to treat this disease. The objective of this study was to evaluate the parameters surviving fraction at 2 Gy (SF(2) ), α/β ratio and capacity for sublethal damage repair (SLDR) in response to radiation. Dose-response and split-dose studies were performed using the clonogenic assay. The mean SF(2) for three established TCC cell lines was high at 0.61. All the three cell lines exhibited a low to moderate α/β ratio, with the mean being 3.27. Two cell lines exhibited statistically increased survival at 4 and 24 h in the dose-response assay. Overall, our results indicate that the cell lines are moderately radioresistant, have a high repair capacity and behave similarly to a late-responding normal tissue. These findings indicate that the radiation protocols utilizing higher doses with less fractionation may be more effective for treating TCC.     

鸟氨酸脱羧酶抗酶1基因的结构和功能

鸟氨酸脱羧酶抗酶1(OAZ1)基因可通过特殊的+1移码机制翻译全长的功能蛋白.研究发现,OAZ1能与鸟氨酸脱羧酶(ODC)结合并降解ODC,负调控细胞内多胺的水平;OAZ1还能降解Cyclin D1、Cyclin E1和Smad1周期蛋白,阻滞细胞周期;此外,近年来研究表明,OAZI还具有抗肿瘤效应和调控动物繁殖的功能.抗酶抑制因子能竞争性结合ODC-OAZ1复合体中的OAZ1,从而阻止ODC降解;天门冬酰胺也能通过抑制OAZ1的翻译来调节ODC的活性.本文就OAZ1基因结构和功能的研究现状作一综述.     

湖南省山羊脑多头蚴的线粒体nad1和nad4基因的序列测定及种系发育分析

本研究旨在阐明脑多头蚴湖南分离株线粒体烟酰胺腺嘌呤二核苷酸( NADH)脱氢酶亚单位1基因(nad1)部分序列(pnad1)和烟酰胺腺嘌呤二核苷酸(NADH)脱氢酶亚单位4基因(nad4)部分序列(pnad4)的遗传变异情况,并用pnad1和pnad4序列重构脑多头蚴与其它带科绦虫的种群遗传关系.利用聚合酶链反应(PCR)扩增脑多头蚴的pnad1和pnad4,应用ClustalX 1.81程序对序列进行比对,再用Phylip3.67程序MP法和Mage4.0程序NJ法绘制种系发育树,并用Puzzle5.2程序构建最大似然树,同时利用DNAstar5.0中的Megalign程序进行同源性分析.结果显示所获得的pnad1和pnad4序列长度分别均为666和887 bp,湖南分离株与已知多头带绦虫位于同一分枝.由于脑多头蚴pnad1和pnad4序列种内相对保守,种间差异较大,故均可作为种间遗传变异研究的标记,从而为脑多头蚴的分子流行病学和其相关疾病的诊断奠定基础.     

胰岛素受体-1在尼罗罗非鱼不同组织中的表达及其对注射葡萄糖的响应

本试验旨在研究胰岛素受体-1(IR-1)在尼罗罗非鱼不同组织中的表达及其对注射葡萄糖的响应。利用PCR扩增的方法从尼罗罗非鱼肌肉中克隆IR-1的c DNA片段,并通过半定量PCR检测,比较IR-1在肌肉、肝脏和心脏中的表达差异。选取体重约为100 g的尼罗罗非鱼160尾,随机分2组,每组4个重复,每个重复20尾。试验组腹腔注射葡萄糖(每100 g体重30 mg),对照组以相同剂量腹腔注射0.7%的无菌生理盐水。在注射前(0 h)和注射后的1、3、6和12 h分别进行采样,测定血浆葡萄糖和胰岛素含量,并通过实时荧光定量PCR检测IR-1在肌肉、心脏和肝脏中的mRNA相对表达量。结果显示:1)克隆出的IR-1 c DNA片段,其Gen Bank登陆号为JN967750,大小为1 979 bp,编码548个氨基酸。序列分析发现,尼罗罗非鱼的IR-1与其他物种比较具有很高的保守性,并具有丰富的酪氨酸激酶特征性序列。2)IR-1在尼罗罗非鱼肌肉、心脏和肝脏中均有较高的表达量,其中在肝脏和肌肉中的表达量基本一致,而在心脏中的表达量相对较低。3)试验组血浆葡萄糖含量在注射葡萄糖1 h后达到最高,并显著高于对照组(P0.05),而后开始下降,3 h后恢复到正常水平;试验组血浆胰岛素含量在葡萄糖注射3 h后达到最高,并显著高于对照组(P0.05),而后开始下降,12 h后恢复到正常水平。试验组肌肉和肝脏中IR-1 mRNA的相对表达量在注射葡萄糖后6 h时达到最高,显著高于对照组(P0.05),在12 h时恢复到正常水平;试验组心脏中IR-1 mRNA的相对表达量在注射葡萄糖后的12 h内没有发生显著变化(P0.05)。结果表明,注射葡萄糖后即刻升高了尼罗罗非鱼的血浆葡萄糖含量,相对于血浆葡萄糖含量的升高,血浆胰岛素含量的升高相对延迟,而肌肉和肝脏中IR-1 mRNA相对表达量的提高又延迟于血浆胰岛素含量的升高,从而加重了尼罗罗非鱼对葡萄糖的代谢负担。     

猪流感病毒A/swine/Guangdong/2/2012(H1N1)毒株攻毒BALB/c小鼠细胞因子动态变化的研究

为了测定H1N1亚型猪流感病毒(swine influenza virus,SIV)对小鼠的致病性,本试验对A/swine/Guangdong/2/2012(H1N1)株SIV HA基因进行克隆及遗传分析,并将SIV尿囊液经鼻腔感染6周龄BALB/c小鼠,观察感染后小鼠的一般状况、器官系数和组织病理学变化,在病毒感染后第1、3和7天使用荧光定量PCR测定小鼠肺脏、脾脏、脑组织中7种细胞因子mRNA的表达量,研究其对小鼠的致病特性。结果显示,该病毒属于经典SIV,病毒经鼻腔感染后可引起小鼠活动减少、采食量降低,但无咳嗽和死亡;病理组织学变化为肺间隔较正常组织明显增厚,毛细血管明显扩张充血,周围肺泡腔呈代偿性肺气肿;小鼠肺脏、脾脏组织样本中IFN-α、IFN-β、IP-10、IL-1β、TNF-α、IRF-3和IL-10 mRNA含量在感染后第3天均显著升高(P<0.05),而脑组织样本中IL-1β和IL-10在小鼠攻毒后第3和7天均显著上调(P<0.05)。