大黄鱼肿瘤抑制因子cylindromatosis (LcCYLD)的序列与免疫反应特征及其功能

为研究大黄鱼肿瘤抑制因子cylindromatosis（CYLD）在免疫反应中的作用,本实验克隆了大黄鱼CYLD的全长cDNA（命名为LcCYLD）并对其进行序列分析;采用荧光定量PCR的方法对大黄鱼各组织及免疫刺激后的大黄鱼肾细胞系中的LcCY LD表达变化进行检测;构建了重组表达载体pTurboGFP-CYLD及pcDNA3.1-CYLD,分别用于亚细胞定位实验及过表达实验;在HEK293T细胞系中过表达LcCYLD后采用双荧光素酶报告系统检测了 NF-κB、TNF-α及IL-1β 启动子活性的变化。结果显示,LcCYLD的ORF包含2 754 bp,编码917个氨基酸,推测具有保守的3个N端的CAP-GLY结构域,1个磷酸化区域和1个C端的UCH结构域,多序列比对结果显示各物种CYLD间高度保守;系统进化分析显示,LcCYLD与来源于其他硬骨鱼的CYLD聚为一支,其中与条纹鲈的CYLD关系最近;转录水平表达分析发现LcCYLD在大黄鱼各组织均有表达,其中在脑中表达量最高;LPS及poly I:C刺激能够显著诱导LcCYLD的表达;亚细胞定位实验表明LcCYLD在细胞质及细胞核中均有表达;过表达LcCYLD能够显著抑制NF-κB及促炎细胞因子TNF-α及IL-1β的转录激活。以上研究结果表明大黄鱼CYLD能够抑制NF-κB的转录激活,为深入了解LcCYLD在大黄鱼先天免疫信号转导中的作用奠定基础。     

牛食道阻塞防治

牛食道阻塞又被称为食道梗塞,是因为食道中被大块食物堵塞,难以下咽所引发的一种急性食道疾病。按照阻塞程度的不同划分为完全阻塞和不完全阻塞2种。按照阻塞部位的不同可以划分为颈部食道阻塞、胸部食道阻塞和腹部食道阻塞3种。因阻塞性质和阻塞程度不同,会继发不同程度的瘤胃臌气。牛食道阻塞具有发病急、发病突然、发病过程快、致死率高的特点,发生后如果不能立即采取措施抢救,患病牛在短时间内会死亡。为提高食道阻塞救治成功率,该文主要探讨牛食道阻塞的诊断和防治过程。     

碳水化合物种类和水平对大黄鱼生长性能、血清生化指标、肝脏糖代谢相关酶活性及肝糖原含量的影响

本试验旨在研究碳水化合物种类和水平对大黄鱼生长性能、全鱼和肌肉常规成分、血清生化指标、肝脏糖代谢相关酶活性及肝糖原含量的影响。采用2×3双因素试验设计,选取葡萄糖和小麦淀粉2种碳水化合物,分别设0、15%、30%3个碳水化合物水平,共配制5种等氮等脂试验饲料。每种饲料饲喂3个重复,每个重复放养初始体重为(8.53±0.07)g的大黄鱼50尾,养殖试验持续8周。结果表明:饲料碳水化合物种类和水平及其交互作用对大黄鱼的终末体重、增重率(WGR)、特定生长率(SGR)、饲料系数(FCR)有显著影响(P0.05)。饲料葡萄糖水平由0增加到30%时,大黄鱼的终末体重、WGR和SGR显著降低(P0.05);饲料小麦淀粉水平由0增加到15%时,大黄鱼的终末体重、WGR和SGR显著升高(P0.05),饲料小麦淀粉水平由15%增加到30%时,大黄鱼的终末体重、WGR和SGR显著降低(P0.05)。FCR随饲料葡萄糖水平的升高显著升高(P0.05);FCR在饲料小麦淀粉水平由0增加到15%时显著降低(P0.05),由15%增加到30%时显著升高(P0.05)。15%或30%水平下,小麦淀粉组大黄鱼的终末体重、WGR和SGR均显著高于葡萄糖组(P0.05),饲料系数显著低于葡萄糖组(P0.05)。饲料碳水化合物种类和水平的交互作用对大黄鱼血清总蛋白(TP)、总胆固醇(TC)、甘油三酯(TG)、葡萄糖含量有显著影响(P0.05)。血清葡萄糖含量随饲料葡萄糖的水平升高先降低后升高,各组间差异显著(P0.05);血清葡萄糖含量随饲料小麦淀粉水平的升高逐渐降低,且30%小麦淀粉组显著低于0小麦淀粉组(P0.05)。饲料碳水化合物种类和水平的交互作用对大黄鱼肝脏葡萄糖激酶(GK)、6-磷酸果糖激酶(PFK)、葡萄糖-6-磷酸酶(G6Pase)活性及肝糖原含量有显著影响(P0.05)。肝糖原含量随饲料葡萄糖的水平升高持续升高,而随饲料小麦淀粉水平的升高先升高后降低,且15%和30%水平下葡萄糖组均显著高于小麦淀粉组(P0.05)。由此得出,与葡萄糖相比,摄食含小麦淀粉饲料的大黄鱼能够通过调节糖代谢相关酶活性及肝糖原含量来维持血糖浓度的相对恒定,且饲料中添加15%小麦淀粉时能促进大黄鱼生长。     

猪CstB基因3’侧翼序列的克隆研究

为了获得猪Cst B基因3’侧翼序列,试验以大白猪cDNA为模板,采用RACE克隆方法,根据GenBank上提供的猪外显子3序列设计引物。结果表明:猪Cst B基因3’侧翼序列长度为225 bp,并提交GenBank收录(GenBank登录号为EF483823)。     

大规格西洋杜鹃嫁接快繁技术

从砧木的采集、处理,接穗选择,嫁接及嫁接后管理等技术,对大规格西洋杜鹃嫁接育苗技术进行总结。     

Genetic characterization of 2 novel feline caliciviruses isolated from cats with idiopathic lower urinary tract disease

Feline caliciviruses (FCVs) are potential etiologic agents in feline idiopathic lower urinary tract disease (I-LUTD). By means of a modified virus isolation method, we examined urine obtained from 28 male and female cats with nonobstructive I-LUTD, 12 male cats with obstructive I-LUTD, and 18 clinically healthy male and female cats. All cats had been routinely vaccinated for FCV. Two FCVs were isolated; I (FCV-U1) from a female cat with nonobstructive I-LUTD, and another (FCV-U2) from a male cat with obstructive I-LUTD. To determine the genetic relationship of FCV-U1 and FCV-U2 to other FCVs. capsid protein gene RNA was reverse transcribed into cDNA, amplified, and sequenced. Multiple amino acid sequence alignments and phylogenetic trees were constructed for the entire capsid protein, hypervariable region E, and the more conserved (nonhypervariable) regions A, B, D, and F. When compared to 23 other FCV isolates with known biotypes, the overall amino acid sequence identity of the capsid protein of FCV-U1 and FCV-U2 ranged from 83 to 96%; identity of hypervariable regions C and E ranged from 58 to 85%. Phylogenetically, FCV-U1 clearly separated from other FCV strains in phenograms based on nonhypervariable regions. In contrast, FCV-U2 consistently segregated with the Urbana strain in all phenograms. Clustering of isolates by geographic origin was most apparent in phenograms based on nonhypervariable regions. No clustering of isolates by biotype was apparent in any phenograms. Our results indicate that FCV-UI and FCV-U2 are genetically distinct from other known vaccine and field strains of FCV.     

丹东地区大果杂交榛子整形修剪技术

本文对丹东地区栽植大果杂交榛子树整形修剪技术进行实践与探讨,以期为丹东地区榛子产业快速发展提供借鉴。     

p38MAPK expression and apoptosis in the rat unilateral ureteral obstruction model

AIM: To investigate the expression of p38 mitogen-activated protein kinase (p38MAPK) in the kidney after unilateral ureteral obstruction (UUO) in rats and the functional role of it on apoptosis and fibrosis.METHODS: Eighteen Wistar rats underwent UUO were killed at 3, 7, 14 days. Additional 7 rats were sham operated. Histological changes were observed by HE and Masson staining. Immunohistochemistry study was performed on renal tissue for proliferating cell nuclear antigen (PCNA). Apoptotic cells were determined by terminal deoxynucleotidyl transferase- mediated dUTP nick end labeling (TUNEL) and the electrophoresis analysis of genomic DNA. Western blotting of cysteinyl aspartate specific proteinase-3 (caspase-3), p38MAPK and p-p38MAPK were measured.RESULTS: UUO induced a significant increase in renal tubular and interstitial cell apoptosis, immunohistochemistry of PCNA and Western blotting of caspase-3, p-p38MAPK as well as severe morphology changes. However, there was no significant difference between UUO and the control in Western blotting of p38MAPK.CONCLUSION: An in vivo model of renal fibrosis after UUO demonstrates that activated or phosphorylated p38MAPK plays a role in apoptosis of renal tubulointerstitial cells.