鸡贫血病毒安徽分离株感染性克隆的构建及其感染特性

鸡贫血病毒（chicken anemia virus,CAV）是诱导雏鸡发生免疫抑制的重要病原之一。应用PCR方法从安徽某鸡场送检病鸡肝组织中检测并鉴定CAV,扩增病毒全基因组序列插入pcDNA3.1(+)载体,并引入含Kpn I （GGTACCCAG）酶切位点的9 bp外源性标签,获得顺次连接的双拷贝CAV基因组的重组质粒,通过脂质体介导转染鸡MDCC-MSB1淋巴细胞并拯救出CAV感染性克隆毒株。经动物试验,感染性克隆毒株在接种1日龄健康雏鸡后15 d,对胸腺等免疫器官指数影响较小,胸腺病变不明显,表明其感染能力减弱,为进一步分析CAV致病机制和筛选疫苗候选株奠定基础。     

汉源县加强天然林资源保护工程的对策措施

天然林是最稳定的生态系统,也是生物多样性的重要基因库。文章在总结汉源县天然林资源保护成效的基础上,分析了存在的森林管护、商品林采伐、公益林投资标准和经果林发展占用林地等问题,并提出了加强天然林资源保护工程的对策措施。     

枇杷果实EjNADP-ME2基因及启动子克隆与表达分析

以‘解放钟’和‘白梨’果实为材料,研究基于转录组筛选出的表达量较高且在2个品种中有差异的EjNADP-ME2（Unigene0001461）基因,采用RACE和RT-PCR技术成功克隆EjNADP-ME2的cDNA全长。系统进化树分析表明,EjNADP-ME2与苹果的NADP-ME基因关系最近。生物信息学预测结果显示,EjNADP-ME2蛋白应该位于细胞质中。用染色体步移法成功克隆了EjNADP-ME2的启动子序列,序列预测该基因启动子区域含有水杨酸、茉莉酸、脱落酸响应的顺式作用元件,此外还有厌氧诱导元件、MYB和MYC参与黄化和干旱诱导的结合位点MBS。荧光定量PCR分析表明,EjNADP-ME2基因的表达量变化趋势在2个品种中大致相同,均呈现出先降低后升高又降低的趋势,且该基因在低酸品种的表达量稍低于高酸品种。相关性分析结果显示,低酸品种EjNADP-ME2基因转录水平与苹果酸含量呈显著负相关。本研究的结果为进一步探究EjNADP-ME2基因在苹果酸代谢途径中的功能奠定坚实基础。     

陕西省商州区地质灾害成因及防治研究

地处陕两省东南部的商州区是一个群山连绵．沟壑纵横,以中低山为主的土石山区,属于地质灾害易发地区,由于自然和人类工程——经济活动的诱发,水土流失比较严重,崩塌、滑坡、泥石流、地面塌陷等地质灾害时有发生。在调查分析该区地质地貌、土壤、植被、水源条件、人类活动等地质灾害成因的基础上,针对商州区实际提出了统筹规划,建立科学的防治预案,提高环境意识,加强植被保护,建设生态商州,加强土地资源的管理和可持续利用,加强矿区综合治理等防灾减灾的对策建议。     

沙苑子对四氯化碳所致大鼠慢性肝损伤的保护作用

目的观察沙苑子对四氯化碳(CCl4)所致大鼠慢性肝损伤的保护作用。方法 SD雄性大鼠60只,随机分成正常对照组(A组),模型组(B组),姜黄素阳性对照组(C组),沙苑子组(D组)各15只。A组皮下注射精制花生油,其他各组采用40%CCl4油剂制备慢性肝损伤模型。6周后,C组、D组分别灌胃给予姜黄素水溶液(20mg/ml)和沙苑子水煎液(30mg生药/ml),A、B组则给予等量的蒸馏水。同时,除A组外,各组仍每周注射40%CCl4油剂1次,持续4周。观察肝脏病理,检测大鼠血清ALT、AST、ALP及肝匀浆MDA含量。结果与B组比较,A组大鼠的肝指数,血清ALT、AST、ALP显著降低,C、D组大鼠肝脏的相对重量、ALT、AST、ALP、MDA均显著降低(P0.01)。结论沙苑子通过降低ALT、AST、ALP及MDA含量发挥抗慢性肝损伤的作用。     

火龙果皮与苜蓿、精料配比对饲料组合效应的研究

本试验旨在探讨火龙果皮与苜蓿、精料配比对饲料组合效应(AE)的影响。试验采用体外产气法测定在精粗比分别为40∶60和30∶70时,精料∶火龙果皮∶苜蓿分别为40∶60∶0、40∶45∶15、40∶30∶30、40∶15∶45、40∶0∶60和30∶70∶0、30∶55∶15、30∶40∶30、30∶25∶45、30∶10∶60、30∶0∶70时11种饲料组合及3种单独饲料(精料、火龙果皮、苜蓿)分别培养0、2、4、6、9、12、24、36、48 h的产气量(GP),培养结束后的上清液及残渣用以测定pH、总挥发性脂肪酸(TVFA)与氨态氮(NH3-N)含量、干物质降解率(DM D)和有机物降解率(OM D),并计算出各指标的单项组合效应指数(SFAEI)和综合组合效应指数(MFAEI)。结果表明:精粗比为40∶60时,40∶60∶0、40∶45∶15组24 h产气量(GP24 h)、DMD、OMD的SFAEI显著或极显著高于其他各组(P<0.05或P<0.01),40∶60∶0、40∶45∶15、40∶30∶30组TVFA和NH3-N的SFAEI显著或极显著高于其他各组(P<0.05或P<0.01),40∶60∶0、40∶45∶15组的MFAEI极显著高于其他各组(P<0.01)。精粗比为30∶70时,30∶70∶0组GP24 h的SFAEI极显著高于其他各组(P<0.01),30∶70∶0、30∶55∶15、30∶40∶30组DM D和TVFA的SFAEI极显著高于其他各组(P <0. 01),30∶70∶0、30∶55∶15组OM D和NH3-N的SFAEI极显著高于其他各组(P <0. 01),30∶70∶0、30∶55∶15、30∶40∶30组的M FAEI极显著高于其他各组(P<0.01)。由此可见,精粗比为40∶60时,精料∶火龙果皮∶苜蓿为40∶45∶15时的M FAEI最大;精粗比为30∶70时,精料∶火龙果皮∶苜蓿为30∶70∶0时的M FAEI最大。     

抗猪IgG单克隆抗体的制备与应用

采用辛酸一硫酸铵盐析和SuperdexTM-200 凝胶层析分离纯化猪血清IgG,免疫BALB/c小鼠,利用细胞融合技术获得8株稳定分泌抗猪IgG 单克隆抗体的杂交瘤细胞.Ig 亚类分析表明,所制备单克隆抗体的轻链亚类均为K,重链除6H3为IgG2b外,其余7株均属于IgG1.Western blot分析显示,5株单克隆抗体识别猪IgG轻链,3株单克隆抗体识别猪IgG重链.以蛋白G亲和层析纯化单克隆抗体7H1腹水,用改良过碘酸钠法进行辣根过氧化物酶(HRP)标记,结果显示,HRP 标记抗猪IgG单克隆抗体对纯化猪IgG的工作浓度达1:12 800.将HRP标记鼠抗猪IgG单克隆抗体与HRP标记羊抗猪IgG多克隆抗体应用于猪血清圆环病毒抗体检测,二者符合率为95.12%,其ELISA和Westem blot的工作浓度为1:2 000和1:10000,表明HRP标记抗猪IgG单克隆抗体具有高度的敏感性.本研究制备的酶标抗猪IgG单克隆抗体为猪病的免疫诊断试剂研究和猪细胞生物学研究提供了一种基础工具.     

玉米芯与苜蓿、精料配比对饲粮组合效应的影响

为了探讨玉米芯与苜蓿、精料间不同配比对饲粮组合效应(associative effects, AE)的影响。采用体外产气法测定在精粗比40∶60和30∶70时,精料∶玉米芯∶苜蓿分别为40∶60∶0、40∶45∶15、40∶30∶30、40∶15∶45、40∶0∶60和30∶70∶0、30∶55∶15、30∶40∶30、30∶25∶45、30∶10∶60、30∶0∶70时各饲料组合及各种单独原料分别培养2,4,6,9,12,24,36,48 h的产气量(gas production, GP),培养结束后的上清液及残渣用以测定pH、挥发性脂肪酸(volatile fatty acids,VFA)、氨态氮(NH_3-N)、干物质降解率(dry matter digestibility, DMD)和有机物降解率(organic matter digestibility, OMD),并通过各组合的加权估算值计算出各指标的单项组合效应指数(single factor AE indexes, SFAEI)和综合组合效应指数(multiple-factors AE index, MFAEI)。结果表明:当精粗比为40∶60和30∶70时,玉米芯比例为30、15和10、25、40、55的组在GP、DMD、OMD、NH_3-N、TVFA上显著高于其他组,且有较高的SFAEI (P<0.05,P<0.01),30和10组的MFAEI极显著高于其他各组(P<0.01)。结论:当精料∶玉米芯∶苜蓿为40∶30∶30、30∶10∶60时,饲粮的综合组合效应值最大。     

北京鸭肌肉生长抑制素基因多态与屠体性状关系分析

肌肉生长抑制素（Myostatin,MSTN）基因在发育和成熟的骨骼肌中特异表达,对肌肉具有负调控作用。应用聚合酶链式反应单链构象多态性（PCR-SSCP）的方法,对6周龄北京鸭2个群体（Z2系和Z4系）共200个个体Myostatin基因5’调控区进行SNP分析,并对所发现的单核苷酸多态性与胴体重、胸肌重、腿肌重、全净膛重、皮脂重、腹脂重、骨架重、胸肌率、腿肌率、皮脂率、腹脂率进行了关联分析。结果发现,A1/A2位点基因型（AA和AB型）对6周龄Z2系和Z4系北京鸭的腹脂重（P＜0.05）和胸肌重（P＜0.10）有显著影响,因此可以将Myostatin基因作为影响北京鸭Z2系和Z4系腹脂重和胸肌重的候选基因。     

Effect of astragalus polysaccharides on expression of MMP2 and MMP9 in annulus fibrosus of cervical intervertebral discs from cervical spondylosis model rats

AIM: To investigate the effect of astragalus polysaccharides (AP) on the expression of matrix metalloproteinase 2 (MMP2) and MMP9 in the annulus fibrosus of cervical intervertebral discs from cervical spondylosis model rats. METHODS: The model rats were randomly divided into model group (M group), and low-dose and high-dose AP treatment groups (L-AP and H-AP groups). The rats in sham operation group were used as negative control group (NC group). In addition, all the annulus fibrosus tissues were used for primary cell culture. Histological analysis was performed using HE staining and Safranin O staining. The expression of MMP2, MMP9, tissue inhibitor of metalloproteinase 2 (TIMP2) and collagen Ⅳ at mRNA and protein levels was analyzed by immunohistochemistry, Western blot and RT-qPCR. Cell-collagen adhesion assay was used to detect annulus fibrosus cell-collagen adhesion. RESULTS: The intervertebral discs of M group were degenerated, while astragalus polysaccharide improved the degenerative disc disease in the rats with cervical spondylosis. Compared with NC group, the expression of MMP2 and MMP9 in the annulus fibrosus tissues of M group increased significantly, while the expression of TIMP2 and collagen Ⅳ was decreased significantly (P<0.05). Compared with M group, the expression of MMP2 and MMP9 in L-AP group and H-AP group was significantly decreased, while the expression of TIMP2 and collagen Ⅳ was significantly increased (P<0.05). The cell-collagen adhesion in M group was significantly lower than that in NC group (P<0.05). Compared with M group, the cell-collagen adhesion in L-AP group and H-AP group was increased significantly (P<0.05). Compared with NC group, the expression of MMP2 and MMP9 in annulus fibrosus cells of M group was increased significantly, while the expression of TIMP2 and collagen Ⅳ was decreased significantly (P<0.05). Compared with M group, the expression levels of MMP2 and MMP9 in L-AP group and H-AP group of fibrocytes were significantly decreased, while the expression of TIMP2 and collagen Ⅳ was significantly increased (P<0.05). CONCLUSION: Astragalus polysaccharides inhibit the expression of MMP2 and MMP9 in the annulus fibrosus of cervical intervertebral discs from cervical spondylosis model rats and regulate the dynamic balance of MMPs and TIMPs in the extracellular matrix, thus inhibiting the degradation of collagen in the intervertebral disc matrix and having the potential research value for the treatment of intervertebral disc degeneration.