Effect of atorvastatin on neointimal hyperplasia of venous autografts in rats

AIM: To investigate the effect of atorvastatin on neointimal hyperplasia of autogenous vein graft in rats. METHODS: The model of autogenous vein graft was prepared by transplanting the external jugular vein into aorta in Wistar rats. The rats were divided into three groups: sham operation group, graft control group and graft experimental group. From three days after transplantation, the rats of autograft experimental group were treated by atorvastatin at a dosage of 5 mg·kg-1·d-1. Four weeks after treatment, venous autografts were removed at autopsy and cut into 4 μm sections. Histopathological examination was carried out to analysis the neointimal hyperplasia of grafted veins. Immunohistochemical staining was conducted to evaluate SMα-actin and PCNA expression of neointimal cells in venous autografts. RESULTS: In venous autograft control and experimental groups, SMα-actin-positive smooth muscle cells were proliferated and accumulated excessively in venous autografts, which resulted in significant neointimal formation and vascular lumen narrowing. Neointima quantitative assay revealed that the neointimal hyperplasia of venous autografts was suppressed obviously in graft experimental group, and its neointimal area and NIA/MA ratio of venous autografts were significantly lower than those in graft control group (P<0.01). Immunohistochemical assay indicated that the PCNA labeling index of neointimal cells was significantly lower in graft experimental group than that in graft control group (P<0.01). CONCLUSION: Atorvastatin significantly inhibits the proliferation of neointimal smooth muscle cells and the development of neointimal hyperplasia of venous autografts in rats. Atorvastatin is a powerful inhibitor of restenosis after vascular reconstructive operation with a potential for therapeutic use.     

基于Web的陕北白绒山羊辅助育种管理系统的设计与实现

针对陕北白绒山羊多性状选择育种研究处于起步阶段的现状,以动物模型BLUP法为核心的多性状复合育种技术体系为依托,在Windows XP平台下利用ASP技术和SQL Server 2000数据库开发了一套基于Web的陕北白绒山羊辅助育种管理系统-ABMS。该系统能实时地管理分布在不同地域种羊场的生产性能测定信息,并能方便地导入和导出选择性状的育种数据。ABMS在陕北白绒山羊种羊场实际应用中体现了技术先进、易于操作、网络化管理等特点,其推广和应用对实现种羊场育种工作和生产管理的自动化,具有极大的促进作用。     

Protective effect of ethyl pyruvate against LPS-induced acute lung injury in dogs with septic shock

AIM:To investigate the effect of ethyl pyruvate (EP) on acute lung injury in dogs with septic shock.METHODS:Twenty dogs with septic shock induced by lipopolysaccharide (LPS) were randomly divided into two groups:dog received placebo (Ringers solution; control group, n=8) or ethyl pyruvate in lactated Ringers solution (0.05 g/kg loading dose over 10 min, thereafter 0.05 g· kg-1·h-1 for 12 h; EP treatment group, n=12).The parameters of respiratory mechanics, including lung dynamic compliance (Cdyn), lung total compliance (Ctot), inspiratory airway resistance (Rawi), peak inspiratory pressure (PIP), work of breathing (WOB), and arterial blood gases were monitored every 4 h after basic measurements (pre-LPS).The contents of TNF-α, IL-6 and IL-10 in serum were also detected by ELISA and compared between two groups.RESULTS:The changes of respiratory mechanics and arterial blood gases were in line with acute lung injury after septic shock induced by LPS.Oxygen index (OI), HCO-3and pH dropped 4 h after septic shock.On the contrary, PCO2 elevated significantly compared with that of septic shock before (P<0.05).Cdyn, Ctot decreased and Rawi, PIP, WOB increased significantly 8 h after septic shock (P<0.05).The changes in parameters of respiratory mechanics and arterial blood gases in control group were more distinguished than those in EP treatment group.There were significantly differences between two groups 8 h after septic shock (P<0.05).Although the significant improvement of respiratory mechanics was not observed, EP did prevent the parameters from worsening further more.Significantly differences between the two groups 12 h after septic shock (P<0.05) were found.Ethyl pyruvate also regulated the network of cytokines:increased the level of TNF-α, IL-6 and reduced the content of IL-10 in serum significantly compared with control group (P<0.05).CONCLUSION:Ethyl pyruvate has protective effects on acute lung injury in dogs with septic shock.     

48kW拖拉机离合器设计与分析

进行了48 kW拖拉机离合器的设计并进行了结构改进:增加一套副离合器操纵系统,节省驾驶员的体力;开有梯形槽的碟形弹簧在制造过程中易产生应力集中,把梯形槽改为弧形槽使离合器寿命大有提高。     

甘德县草原使用权流转中的问题及对策

综合甘德县草原使用权流转的形式，分析了流转的原因、作用及存在的问题，并提出了甘德县草原使用权流转的对策建议。     

用厨余食物生物发酵饲料饲养肉鸡试验报告

将餐厅、宾馆、饭店、食堂等厨房加工菜肴的“边角余料”和餐桌的残余食物通过专用加工机械进行灭菌、生物发酵、干燥等工艺流程加工成饲料，作者将本饲料按不同配比添加在15～56日龄肉鸡饲料中，与常规饲料配方饲养进行对比试验。试验结果表明，试验组与对照组在肉鸡生长发育和鸡肉的品质等方面差异不显著，证明本饲料可以替代部分肉鸡浓缩料和玉米面，使厨余食物不但变废为宝，降低了饲养成本，还保护了环境。     

TCDCA对有丝分裂原刺激的小鼠脾淋巴细胞免疫功能的影响

为了探讨牛磺鹅去氧胆酸(TCDcA)对有丝分裂原刺激的小鼠脾淋巴细胞免疫功能的影响,设计了TCDCA的0.01,0.05,0.1,1,10μg/mL5个剂量浓度梯度,开展了TCDCA对有丝分裂原刺激的小鼠脾淋巴细胞增殖反应、分泌IL-2和IgG的影响研究。结果表明:TCDCA对ConA刺激的脾淋巴细胞增殖反应表现出调节作用,对LPS刺激的脾淋巴细胞无明显作用;TCDCA对ConA和LPS刺激的脾淋巴细胞的分泌IL-2表现出向上抛物线形式的明显的调节作用;TCDCA对LPS刺激的脾淋巴细胞分泌IgG具有向下抛物线形式的明显的调节作用。说明TCDCA对有丝分裂原刺激的小鼠脾淋巴细胞免疫功能具有明显的调节作用。     

基于转录组数据分析环形泰勒虫对诱导宿主细胞转化相关基因的影响

本试验主要通过研究布帕伐醌(BW720)处理前后,环形泰勒虫转化牛单核细胞(TaNM Ⅰ)转录组数据的变化情况,为进一步研究环形泰勒虫诱导宿主细胞转化的分子机制奠定基础。以环形泰勒虫感染的牛单核细胞为试验材料,设置对照组(二甲基亚砜,DMSO)和试验组(BW720),利用Illumina HiSeq4000测序平台进行高通量测序,将测序得到的原始数据(Raw reads)与GenBank和Rfam数据库进行比对过滤,通过质控获得最终数据(Clean read)。利用Venn分析软件筛选出DMSO组和BW720组中差异表达明显的基因,进行功能注释(GO)以及信号通路(KEGG)分析。随机选择10个基因,利用荧光定量PCR (qPCR)检测基因在不同处理条件下细胞中的表达量。在验证测序结果的准确性后,选出差异表达明显的基因,分析其在TaNM Ⅰ细胞的凋亡、增殖等信号通路方面的作用。结果显示：BW720组和DMSO组中分别得到6 854 019 704和6 627 265 854条Raw reads,过滤、质控后分别得到22 925 606和22 171 427条Clean reads。BW720组和DMSO组中筛选出差异表达明显的基因共计4 054个(P＜0.05),其中,2 146个基因显著上调,1 908个基因显著下调;进一步筛选出共同差异表达的基因367个,其中,显著上调的196个,显著下调的171个。同时,qPCR验证随机选择10个基因的表达趋势与转录组测序结果一致,表明测序结果可靠。然后对差异表达基因进行GO功能注释,筛选到与细胞增殖相关的20个生物学过程、15个细胞组分以及11个分子功能条目。KEGG富集分析结果显示,在Top20的信号通路中筛选出与环形泰勒虫转化细胞有关的一些信号通路,如：癌症信号通路、PI3K-Akt信号通路、MAPK信号通路等。对这些信号通路进一步分析发现：PI3KR3、FOXO1、IL23A、FZD3、AKT、MMP9等基因在环形泰勒虫转化细胞的研究中也有相关文献的报道。本研究表明,在TaNM Ⅰ细胞中DAPK1、FZD3、FOXO3、PI3KR3等可能在感染细胞无限增殖的过程中发挥作用,为后续研究TaNM Ⅰ细胞无限增殖机制奠定了基础。     

西门塔尔牛胰腺间充质干细胞原代培养及其多向分化潜能研究

旨在对西门塔尔牛胰腺间充质干细胞（pancreatic mesenchymal stem cells,PMSCs）进行原代培养并研究其体外分化潜能,为细胞疗法和组织工程学方面提供新的种子细胞。本研究从3月龄的西门塔尔牛胚胎中无菌分离胰腺组织,分别采用胶原酶消化法和组织块贴壁法分离PMSCs,进行原代培养;绘制第3代、第9代、第15代细胞生长曲线并测定群体倍增时间及克隆形成能力,采用免疫荧光检测干细胞表面标志物（CD29、CD44、CD73、CD90、CD34和CD45）,RT-PCR检测干细胞细胞表面标志物（CD29、CD44、CD73、CD90、CD106、CD166、CD34和CD45）;染色体核型分析检测其基因稳定性,通过向成脂、成骨、成软骨和成肝样细胞诱导分化,检测其多向分化潜能。结果表明,两种方法都可成功分离出PMSCs,细胞贴壁后形态均为长梭形,漩涡状生长,生长趋势呈典型的S形;第9代细胞群体倍增时间显著低于第15代而高于第3代（P<0.01）;第9代PMSCs克隆形成率显著低于第3代而显著高于第15代（P<0.05）;免疫荧光结果表明,PMSCs特异性表达CD29、CD44、CD73和CD90,RT-PCR结果显示PMSCs特异性表达CD29、CD44、CD73、CD90、CD106和CD166,未表达造血细胞表面标志物CD34和CD45,与国际细胞治疗学会组织干细胞委员会指定的MSCs表面标记物相对应;核型分析表明,PMSCs为正常二倍体（2n=60,XY）,染色体基因组未发生变异;特异性染色和RT-PCR结果表明,从西门塔尔牛体内获得的PMSCs可分化为脂肪细胞、骨细胞、软骨细胞和肝样细胞。本试验证实两种方法均可成功建立西门塔尔牛PMSCs体外分离培养体系,PMSCs具有活性好、增殖速度快的特点,与MSCs有相似的生物学特性和多项分化的潜能。可为组织工程学提供新的种子细胞。     

广灵驴FTO基因克隆、序列分析及组织差异表达

[目的] 对广灵驴脂肪和肥胖相关基因（fat mass and obesity-associated gene,FTO）进行克隆及序列分析,同时检测其在广灵驴各组织中的表达差异,以探究广灵驴FTO基因结构对其生理代谢功能的影响。[方法] 运用RT-PCR技术扩增并克隆广灵驴FTO基因CDS序列,进行基因及蛋白质功能分析,同时运用实时荧光定量PCR方法检测FTO基因在广灵驴7种组织（心脏、肝脏、脾脏、肺脏、肾脏、背最长肌及皮下脂肪）中的差异表达。[结果] 广灵驴FTO基因CDS区序列长1 518 bp,编码505个氨基酸,序列提交至NCBI,登录号：MZ169553。广灵驴FTO基因与马、猪、牛、人、羊驼、绵羊和山羊的相似性为99.3%、90.3%、89.5%、90.8%、90.7%、89.2%和89.2%;系统进化树分析发现,广灵驴与马的亲缘关系最近。FTO蛋白分子质量为58.35 ku,理论等电点为5.07,脂肪系数为80.36,不稳定系数为48.82,平均疏水指数为-0.550,为不稳定的酸性亲水蛋白。FTO蛋白无信号肽和跨膜区,主要定位于细胞质中,具有34个磷酸化位点与5个糖基化位点。FTO蛋白二级结构主要以α-螺旋（43.96%）和无规则卷曲（37.82%）为主。实时荧光定量PCR分析发现,FTO基因在广灵驴7个组织中均有表达,其中在肺脏和皮下脂肪中的表达量极显著高于其他组织（P<0.01）,在背最长肌中表达量最低。[结论] 本试验结果可为下一步基因表达与调控脂肪沉积机制及改善驴肉品质的研究奠定基础。