口蹄疫病毒Asial/JS/China/2005株基因组全长感染性克隆的构建

利用长距离RT-PCR技术扩增病毒基因组3'端覆盖口蹄疫病毒Asial/JS/China/2005株近全长的3个片段(共约7.5 kb),并利用单一酶切位点将其分别克隆到pBlueseriptSK+载体上.利用融合PCR扩增到基因组5'端含有15个C碱基的基因片段(约700 bp),并将其连接至pGEM-T载体.最后将这4个片段的阳性克隆装配至剔除T7启动子的低拷贝载体pcDNA3.1/Zeo(+)中构建该病毒株的全长cDNA克隆.以构建的FMDV Asial/JS/China/2005株全长cDNA为模板,使用TTRNA聚合酶在体外转录得到病毒RNA,通过脂质体将其导入BHK细胞获得拯救病毒.对收获的病毒分别用RT-PCR、间接免疫荧光、电子显微镜观察和乳鼠致病性分析结果证实,通过体外转录获得了具有感染性的口蹄疫病毒.该株感染性克隆的构建为深入研究口蹄疫病毒的致病机制及研制新型疫苗等奠定了基础.     

龙牧807苜蓿选育报告

从龙牧803(Medicago sativa L.cv.Longmu 803)苜蓿群体中选择优秀单株,通过无性扦插繁殖,3次混合选择、开放授粉及品种比较试验、区域与生产试验,历经12年育成龙牧807苜蓿(Medicago sativa L.cv.Longmu807)新品种。该品种株型直立,在黑龙江省地区4月中下旬返青,生育期120 d左右,开花期株高75.7~105.3cm,成熟期株高98.5~133.8 cm。2年生干草产量为11739.14 kg·hm^-2,比对照龙牧803苜蓿(9797.99 kg·hm^-2)增产17.10%;种子产量284.60 kg·hm^-2,比对照(230.00 kg·hm^-2)增产18.25%。初花期粗蛋白含量19.38%,适口性好,饲用品质优良。其抗性强,生态适应性广,适宜在黑龙江省各地及毗邻省区推广种植。     

伪狂犬病毒在潜伏感染猪体内的组织分布

潜伏感染是猪伪狂犬病防治和净化工作中的重要障碍之一。本研究用伪狂犬病毒（PRV）gG-/LacZ＋标记毒株感染经过PRV灭活疫苗免疫的PRV阴性仔猪,建立了猪伪狂犬病毒潜伏感染动物模型,再用地塞米松激活PRV在猪体内的潜伏感染。运用免疫组织化学SABC染色法研究了PRV在潜伏感染猪和潜伏感染激活猪体内部分组织的分布情况。结果显示,阳性细胞主要分布在神经系统的大脑、小脑、脑干、三叉神经和视神经以及非神经系统的扁桃体、肺和肾等部位。阳性细胞数量随着攻毒后时间的发展呈现减少的趋势,而注射地塞米松后,阳性细胞数量在上述组织中显著增加。     

Effects of remifentanil on monophasic action potential and transmural dispersion of repolarization in rabbit myocardium

AIM: To study the effect of remifentanil on monophasic action potential and transmural dispersion of repolarization (TDR) in the 3-layer myocardium of isolated rabbit hearts. METHODS: Adult rabbits (n=18, 2.0 ~ 2.5 kg) were used to isolate the hearts for preparing Langendorff perfusion model. The hearts were randomly divided into 3 groups after perfusion with K-H solution for 15 min: the perfusion in control group (C group) continued for 60 min; the hearts in remifentanil group (R group) were perfused with 12 μg/L remifentanil K-H solution for 60 min; the hearts in remifentanil+aminophylline group (RA group) were given 60-min perfusion of 12 μg/L K-H remifentanil+30 mg/L aminophylline. The HR and 3 layers of myocardial monophasic action potential (MAP) in the left ventricular anterior wall were recorded at time points after balanced infusion for 15 min (T₀), and continued perfusion for 15 min (T₁), 30 min (T₂) and 60 min (T₃). The monophasic action potential duration of repolarization at 90% (MAPD₉₀) and the transmural dispersion of repolarization (TDR) were calculated. The early afterdepolarization, delay afterdepolarization and arrhythmia were also observed. RESULTS: In R group, slower HR and prolonger MAPD₉₀ and TDR at T₁~T₃ were observed as compared with those at T₀ (P<0.05). R group showed slower HR and longer MAPD₉₀ and TDR than C group and RA group (P<0.05). CONCLUSION: Remifentanil slows the HR, extends the MAPD₉₀ and increases the TDR, thus being prone to induce reentry. Aminophylline makes HR faster and MAPD₉₀ shorter, thereby reducing the TDR.     

早花忍冬离体快繁技术

以东北地区早春植物早花忍冬的幼嫩茎段为外植体,MS培养基为基本培养基,添加不同的植物生长调节物质,进行了离体快速繁殖技术研究。结果表明:采用2.0%NaClO处理20min的灭菌效果好;诱导早花忍冬侧芽分化的适宜培养基为MS+1.0mg/L 6-BA+0.05mg/L NAA+3.0%蔗糖+0.7%琼脂;增殖培养基为MS+2.0mg/L 6-BA+0.05mg/L NAA+3.0%蔗糖+0.7%琼脂,30d的增殖系数为8;最佳的生根培养基为1/2MS+0.3mg/L IBA+0.3mg/L NAA+1.5%蔗糖+0.7%琼脂,生根率达100%。     

过表达MyBPC1对牛骨骼肌卫星细胞增殖分化的影响

旨在探究肌球蛋白结合蛋白C1（myosin binding protein C1,MyBPC1）对牛骨骼肌卫星细胞增殖与成肌分化的影响,为进一步研究MyBPC1在细胞分化和肌肉发育过程中的调控作用提供依据。本研究利用西门塔尔胎牛原代牛骨骼肌卫星细胞体外诱导成肌分化模型模拟牛骨骼肌的生长发育过程。采用qRT-PCR和Western blot检测MyBPC1的细胞时序表达谱。试验分为两组。在RNA水平每组4个重复,每个重复20 μL;在蛋白水平每组3个重复,每个重复15 μg。采用qRT-PCR和Western blot检测牛骨骼肌卫星细胞转染MyBPC1的过表达效果,并进一步检测细胞增殖期标志因子Pax7、Ki67以及细胞分化期标志因子MyHC、MyOG的表达变化情况,观察牛骨骼肌卫星细胞肌管形成状态。结果,MyBPC1在牛骨骼肌卫星细胞分化前后表达水平存在极显著差异,牛骨骼肌卫星细胞诱导分化后MyBPC1的mRNA和蛋白表达量均极显著高于增殖期（P<0.01）。过表达MyBPC1后,细胞分化形成的肌管数量明显多于对照组,增殖标志因子Pax7的mRNA水平和蛋白表达水平无显著差异,分化标志因子MyHC的mRNA水平和蛋白表达水平极显著高于对照组（P<0.01）。过表达MyBPC1可以促进牛骨骼肌卫星细胞体外成肌分化,为进一步开展MyBPC1对牛骨骼肌卫星细胞的调控机制奠定基础。     

羊源D型多杀性巴氏杆菌感染小鼠脾脏转录组分析

试验旨在探索羊源D型多杀性巴氏杆菌入侵宿主脾脏组织中引发的免疫应答途径。首先利用羊源D型多杀性巴氏杆菌(HN01菌株)感染小鼠,建立羊源D型多杀性巴氏杆菌感染的动物模型;之后利用转录组测序技术获得感染小鼠与正常小鼠的脾脏转录组数据,并使用COG、KOG、eggNOG、GO、KEGG数据库对测序结果中的差异表达基因(differentially expressed genes,DEGs)进行功能注释与分析,同时对于显著富集到关键免疫通路的差异表达基因使用STRING软件和KEGG mapper进行蛋白互作分析,筛选出核心通路中起关键作用的基因;最后选取关键的10个基因进行实时荧光定量RT-PCR验证。转录组分析结果显示,与正常组相比,感染组中筛选出3 380个差异表达基因(P<0.01,log₂|FoldChange|≥0.5),其中1 691个基因上调,1 689个下调。基因功能富集分析结果表明,感染组脾脏中的差异表达基因主要发挥信号转导的功能,其主要参与的生物途径包括细胞因子与细胞因子受体互作通路、趋化因子信号通路、HIF-1信号通路、TNF信号通路。蛋白互作分析筛选出约28个核心差异表达基因,结合实时荧光定量RT-PCR验证后,其中9个基因的表达结果与测序一致,分别是C3、Cd4、Cxcl13、Lck、Gnai1、Grap2、IL-6、Cxcr6及Serping1基因。本研究初步证明了脾脏在抵抗多杀性巴氏杆菌入侵中参与了一系列免疫应答反应,为进一步研究羊源D型多杀性巴氏杆菌与宿主之间相互作用的分子机制奠定理论基础。     

基于WSN的水环境监控系统设计

针对湿地水环境监控需求,采用CC2530芯片设计无线传感器网络终端节点,基于Z-Stack协议栈构建局域自组织无线网络,水体监测传感器的终端节点采集的数据最终汇聚到由CC2531和ARM构建的协调器上,经GPRS与远程上位机客户端建立通信链接并接入互联网络。该系统可以广泛应用于湿地等大范围环境监控,具有成本低、便于维护和节约人力资源等优点。     

阜新庆年家庭农场发展户示

探讨发展家庭农场在发展现代农业、引领农业发展方面、提升农产品市场竞争力方面的重要意义,根据阜新庆年家庭农场的成功经验,从土地流转方式、扶持政策、新型农民培育3个主要方面,探讨发展壮大家庭农场的主要措施,为家庭农场的发展提供启示。     

Study on Clinical Efficacy of Oligosaccharide Sulfate and Dextran Sulfate Against Bovine Mastitis

In order to detect the clinical efficacy of oligosaccharide sulfate(DEAE) and dextran sulfate (DS-4000) against bovine mastitis,56 cows with bovine mastitis were divided into 5 groups (DEAE group,DS-4000 group,DEAE＋penicillin group,DS-4000＋penicillin group and penicillin group),and their breasts were perfusion with different drugs twice one day for 3 days.The effective rates of drugs were observed by clinic symptoms.Milk samples were collected before and after treatment of drugs,and the bacterial negative conversion rate of each group were detected by microbial examination.The results showed that total effective rates of 5 groups were 10.00%,20.00%,58.33%,75.00% and 50.00%,respectively,and the bacterial negative conversion rates were 0,12.50%,50.00%,54.55% and 44.44%,respectively.In conclusion,the curative efficiencies of the polysaccharides against bovine mastitis were weak,but it could be improved by combination with penicillin.