PRRSV浙江株ORF5基因的原核表达与免疫原性分析

根据GenBank中公布的PRRSV基因序列(EU709837),设计一对PCR引物,以浙江分离株为模板,通过RT-PCR扩增出ORF5基因全序列.以PCR技术构建了缺失信号肽及三处跨膜区的重组ORF5基因,命名为g-ORF5.将g-ORF5基因定向克隆到原核表达载体pET-28a中,转化入E.coli菌株BL21.经酶切和测序鉴定后,阳性菌株用1 mol/L IPTG诱导表达,用Ni-NTA纯化表达的重组蛋白.结果表明,重组GP5蛋白在大肠杆菌中获得表达.纯化后经ELISA和Dot-ELISA分析,表达蛋白具有免疫反应性,能与PRRSV阳性猪血清发生特异性反应,而与阴性血清不反应.     

石菖蒲有效成分分析及抗菌作用研究

采用试管预试法和圆形滤纸层析法,初步分析鉴定了石菖蒲的化学成分,测定了其水提取液对兽医常见病原微生物的体外抑制效果及最小抑菌浓度(MIC)、最低杀菌浓度(MBC)值。旨在为石菖蒲在兽医临床及畜牧生产上的开发利用提供依据。     

HPLC-PDA法测定兽药中非法添加四种解热镇痛抗炎类药物

建立了7种兽药中非法添加对乙酰氨基酚、安乃近、地塞米松和地塞米松磷酸钠药物的HPLC-PDA法。采用十八烷基键合硅胶为填充剂,磷酸二氢钠缓冲液（磷酸二氢钠3．0 g加水至1000 mL,加三乙胺1 mL,用氢氧化钠调pH值至7．0±0．2）-甲醇（80∶20）为流动相,梯度洗脱,二极管阵列检测器进行全扫描和240 nm波长测定,并采用峰纯度检查和光谱相似度检查辅助对照品比对方法,对四种目标分析物进行确证。结果显示,4种解热镇痛抗炎药物与其他物质峰分离良好,在测定范围内线性关系良好,平均回收率为73．5％～119．2％,RSD为0．1％～5．8％。本方法准确、可靠、重现性好,可用于兽药制剂中非法添加四种解热镇痛抗炎药物的定性和定量检测。     

日本血吸虫四跨膜孤儿受体(TOR)克隆及其第一个膜外区基因的表达

埃及血吸虫和曼氏血吸虫四跨膜孤儿受体(trispanning orphan receptors,TOR)受体可以结合补体C2a,可能在血吸虫免疫逃避过程中起重要作用。为研究日本血吸虫TOR受体的特性和功能,利用PCR扩增到Sj TOR基因并进行了生物信息学分析;同时克隆了Sj TOR基因N端第一个膜外区(ed1)基因,构建了重组质粒p ET-28a-Sj TOR-ed1,在大肠杆菌BL21(DE3)中诱导表达后用组蛋白亲和层析纯化获得重组蛋白Sj TOR-ed1,Western blot结果显示r Sj TOR-ed1能被日本血吸虫阳性小鼠血清识别。该研究为进一步开展研究日本血吸虫TOR受体的功能提供了基础。     

智能化太阳能密集烤房节能效果研究

智能化太阳能密集烤房由太阳能供热系统、煤燃烧供热系统、装烟系统、通风排湿系统和自动控制系统等部分构成。通过智能化太阳能密集烤房与普通密集烤房的烟叶烘烤试验,结果表明:烘烤下部烟叶,历时118 h,其中3 d晴天,1 d雨天,1 d阴天,最大光照强度13.60×104lux,智能化太阳能密集烤房烘烤一炕烟叶较对照节煤198.6 kg,节煤效果达25.6%;烘烤中部烟叶,历时138 h,均为晴天,最大光照强度15.20×104lux,智能化太阳能密集烤房烘烤一炕烟叶较对照节煤123.2 kg,节煤效果达36.1%;烘烤上部烟叶,历时146 h,其中1 d阴天,其余为晴天,最大光照强度12.80×104lux,智能化太阳能密集烤房烘烤一炕烟叶节煤255.0 kg,节煤效果达35.6%。智能化太阳能密集烤房与对照烘烤各炕次烟叶的耗电量则无显著差异,智能化太阳能密集烤房的节能减排效果主要表现在节煤上。     

南方红豆杉扦插繁殖技术研究--扦插苗与实生苗光合生理特性比较

对南方红豆杉扦插苗和实生苗光合生理特性比较研究结果表明,南方红豆杉扦插苗叶重、叶绿素a和b、叶绿素总量、CO2补偿点及净光合速率、硝酸还原酶活性均小于相应树龄实生苗。     

南方稻田保护性耕作的研究进展与研究对策

就保护性耕作的由来和内涵、国内外发展现状作一综述,在指出我国稻田传统耕作方式存在的主要问题后,着重介绍了南方稻田保护性耕作的主要技术模式:即以免(少)耕、秸秆还田为主的保护性耕作技术和休闲期保护性耕作技术。稻田免(少)耕保护性耕作技术主要包括水稻免耕直播、免耕抛秧、免耕套播和免耕小苗移栽技术;秸秆还田为主的保护性耕作技术主要包括秸秆覆盖免耕栽培水稻和秸秆覆盖免耕旱作技术;休闲期保护性耕作技术主要包括冬春休闲和休闲期绿色覆盖技术。指出这些保护性耕作技术不仅具有传统栽培同样或更高的产量潜力,而且省工、节本、能减少水土流失,保持和改善地力,经济、生态效益显著。分析稻田保护性耕作存在的不足之处,提出发展对策。     

温度和日照与春播小麦品质的关系

 为了研究温度和日照与春播小麦品质的关系,用3个小麦品种于1998～2001年在日喀则、拉萨、林芝、北京和沈阳进行了春播试验。结果表明,小麦一些品质性状在不同地点之间以及不同年度之间差异极大。蛋白质含量与湿面筋含量、降落值间均呈正相关。播种-成熟天数与蛋白质含量呈负相关。蛋白质含量与抽穗-成熟期间的平均日均温和平均日照时数呈正相关,与平均昼夜温差呈负相关。湿面筋含量与同期平均日均温和平均日照时数呈正相关。沉降值、降落值与此期间的平均日照时数均呈正相关。西藏自治区一般不具备发展强筋小麦的条件,应主要发展中筋小麦     

Mitogen-activated protein kinase is involved in the changes of intracellular free calcium concentration induced by wound exudate in cultured rat epidermal stem cells

AIM: To observe the effects of harvested wound exudate on intracellular free Ca²⁺ in epidermal stem cells (ESCs) in vitro, and to investigate the relationship between mitogen-activated protein kinases (MAPKs) signal pathways and Ca²⁺ mobilization in this condition.METHODS: Wound exudate was harvested from the 80 full-thickness wounds produced on both sides of the back in 40 adult Wistar rats. ESCs were isolated, purified from neonatal Wistar rats by referring to the formerly records and binding our ideas. When the cultured cells showed up clone growing, they were divided into five groups as follows: group A: control group (no-treatment); group B: only treatment with wound exudate; group C: treatment with wound exudate and PD98059; group D: treatment with wound exudate and SB203580; group E: treatment with wound exudate, PD 98059 and SB203580. Then, the cells were incubated with fluorescence Ca²⁺ dye fluo-3/AM at 37 ℃ for 30 min, and measured by using laser scanning confocal microscope.RESULTS: The results showed that the fluorescent intensity of group B was higher than that in group A. A phenomenon of calcium oscillation was found in group C and group D. Furthermore, a rapid decrease of fluorescent intensity was observed in the cells that were preincubated with PD98059 and SB203580 at the same time. CONCLUSION: Based on above results, we propose that wound exudate can directly induce an increase in intracellular free Ca²⁺concentrations of ESCs. MAPKs signaling pathway has an important function of feedback regulation for free Ca²⁺ mobilization of ESCs in this condition, and also is capable of affecting the biological behaviour of epidermal stem cells.     

High density lipoprotein subclasses and activities of protein kinase C

AIM: To investigate the relationship between high density lipoprotein (HDL) subclasses and the binding activities of hepatic cell and extra-hepatic cell HDL receptor and intracellular protein kinase C (PKC). METHODS: Using purified pre-β1 HDL and apoE-deficent HDL3 to react with human smooth muscle cells (SMC) and HepG2 cells, then the activities of pre-β1HDL and apoE-deficent HDL3 binding to SMC and HepG2 cells, and the effects of this two HDL subclasses on PKC activities in SMC and HepG2 cells were observed . RESULTS: The results showed that the value of Kd of pre-β1HDL binding to SMC HDL receptor was not only significantly lower than that of apoE-deficient HDL3 (P<0.05), but significantly lower than pre-β1 HDL binding to HepG2 HDL receptor (P<0.05). Cell PKC system was all activated by binding of the two HDL subclasses with SMC HDL receptor, and the effect of pre-β1HDL appeared to be stronger than apoE-deficent HDL3. No change in HepG2 cell PKC activity by binding the two HDL subclasses with HepG2 HDL receptor was observed. CONCLUSIONS: The results indicate that the ability of pre-β1HDL to promote efflux of cholesterol from extra-hepatic cells is stronger than that of apoE-deficent HDL3, and it seems that plasma pre-β1 HDL mainly interacts with extra-hepatic cell HDL receptor, subsequently, activates PKC signal transduction system and promotes the intracellular cholesterol efflux from cells.