鸡Ii结合Rab5a和Rab7b分子的分析

本研究旨在探明鸡恒定链（invariant chain，Ii）与内吞体转运蛋白Rab5a和Rab7b结合的结构域和在细胞内共定位的特征。首先，用PCR和基因突变技术将Ii胞浆区与跨膜区[Ii_{（Cyt-Tra）}]、Ii CLIP （class Ⅱ-associated invariant chain peptide）-三聚体区[Ii_{（CLIP-TRIM）}]和Ii突变体[Ii_{（M81-87aa）}、Ii_{（M91-99aa）}和Ii_{（M81-99aa）}]分别插入pET-32a和pEGFP-C1构建相应的原核和真核重组质粒。其次，将构建的含有绿色荧光蛋白的重组质粒与实验室保存的含有红色荧光Rab5a和Rab7b的重组质粒共转染至人胚胎肾细胞系293 T，观察它们的共定位。将构建的原核重组质粒进行表达和纯化，最后用拉下法和免疫印迹检测Ii与Rab5a和Rab7b的结合域。结果表明，成功构建Ii结构域及Ii突变体的重组质粒。Ii_{（Cyt-Tra）}及Ii突变体均能与Rab5a和Rab7b在细胞内共定位，而Ii_{（CLIP-TRIM）}与空载体却不能。Ii的胞浆区和跨膜区是与Rab5a和Rab7b结合的功能结构域，而不是CLIP与三聚体区。综上所述，鸡Ii与Rab5a和Rab7b共定位和结合的区域是其胞浆区和跨膜区，而不是内质网腔区。这些结果提示Rab分子参与了Ii在胞内细胞器的转运机制，为进一步研究Ii及其载体在细胞内的转运机制和功能提供了新的途径。     

Influence of all-trans retinoic acid on proliferation and differentiation of neural stem cells from new born rat striatum

AIM: To study the effect and mechanism of all-trans retinoic acid (atRA) on neural stem cell (NSCs) proliferation and differentiation from new born Sprague-Dawley rat striatum. METHODS: NSCs were isolated from the brains of new born Sprague-Dawley rat striatum, and the features of cells were characterized by immunofluorescence staining. The effects of different culture medium on cell cycle distribution and proliferation of NSCs were determined by flow cytometry (FCM). The effects of atRA on differentiation of NSCs were determined by immunofluorescence staining and classified count of differentiated cells. RESULTS: FCM assay indicated that atRA inhibited the proliferation of NSCs. The percentage of cells in G0/G1 phase in atRA treatment group was significantly higher than that in control, and the proliferation index (PI) was significantly low. The percentage of neurons differentiated from NSCs in atRA group was 2.5 times of the control group after induced by adding 10% FCS in culture medium. CONCLUSION: atRA counteracts the effects of bFGF on the promotion of mitosis and inhibition of differentiation of NSCs. atRA also promotes NSCs to differentiate into neurons in vitro.     

猪CD3∈分子的cDNA克隆与序列分析

应用RT-PCR和3′RACE技术,本研究从猪外周血淋巴细胞总RNA中扩增克隆了猪CD3ε基因并进行了序列分析。序列分析结果表明,猪CD3εcDNA序列长1241nt,其中5′非翻译区80nt,3′非翻译区570nt,开放阅读框591nt,该开放阅读框编码196个氨基酸残基的CD3ε前体蛋白(无糖基化位点);在猪CD3ε蛋白的胞外区,Cys49、Cys91、Cys108和Cys111为保守性氨基酸残基,它们与CD3ε免疫球蛋白样结构的形成有关。在猪CD3ε蛋白胞浆区,存在免疫受体酪氨酸活化基序(ITAM)、内质网潴留基序和1个SH3结合基序,其中ITAM基序含有2个SH2结合基序和2个潜在酪氨酸磷酸化位点Tyr177和Tyr188,这些基序是猪CD3分子参与T细胞活化信号转导和TCR-CD3复合体装配的结构基础。推导氨基酸序列分析结果表明,猪与人、鼠、狗、牛和绵羊CD3ε蛋白的氨基酸同源性分别为61.2%、58.7%、58.7%、65.1%和65.6%。     

Role of p38MAPK in production of pro-inflammatory cytokines in Kupffer cells from severe acute pancreatitis rats

AIM:To investigate the role of p38 mitogen-activated protein kinase (p38MAPK) signaling pathway in the Kupffer cells (KCs) production of pro-inflammatory cytokines, tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β), in severe acute pancreatitis (SAP) rats.METHODS:Sprague-Dwaley rats were randomized into three groups:①sham operation rats, ②SAP rats, ③SAP rats given the p38 MAPK inhibitor CNI-1493(10 mg/kg, iv). The SAP model was induced by the bili-pancreatic duct infusion with 5% sterile soduim taurocholate solution. Rats from each group were killed at 12 h after sham operation or SAP and Kupffer cells (KCs) were isolated. The mRNA expressions of TNF-α and IL-1β (by quantitative real-time RT-PCR) and p38 MAPK activity (by Western blot analysis) in KCs were examined. The levels of TNF-α and IL-1β in plasma were determined by ELISA.RESULTS:There was a significant acvitation of p38 MAPK in KCs harvested from SAP rats than those from sham operation rats. SAP also promoted the mRNA expressions of TNF-α and IL-1β in KCs and the plasma levels of TNF-α and IL-1β. These events were significantly inhibited by treatment with CNI-1493.CONCLUSIONS:p38 MAPK activation is one important aspect of the signaling events that may mediate the KCs production of pro-inflammatory cytokines, TNF-α and IL-1β, in SAP rats. The inhibition of the p38 MAPK may be a potential target in the prevention and treatment of SAP.     

全球气候暖干化对秦岭南北河流径流泥沙的影响研究

以秦岭南侧汉江和北侧渭河多年的径流泥沙观测资料，分析了全球气候暖干化对秦岭南北径流泥沙的变化。分析指出在80年代后，由于全球气候变化的影响，秦岭南北河流年均径流量均减少，与1935－1980年相比，汉江河流年均径流量减少1.9%，渭河河流年均径流量减少27.4%；同时汉江河流泥沙含量明显减少，但渭河河流泥沙含量呈增加趋势，是汉江河流泥沙含量133倍，表明了秦岭南北两侧在全球气候暖干化表现出明显的区域响应性。     

猪繁殖与呼吸综合征病毒缺失变异株的基因组特征

对猪繁殖与呼吸综合征病毒(PRRSV)分离株HB2(sh)／2002的全基因组序列进行了测定与分析。该毒株基因组全长为15373nt(不包括PolyA尾)，与国内外美洲型PRRSV分离株全序列相似性介于88．7％～95．1％之间。序列分析表明，该毒株是1个天然存在缺失的变异毒株，其ORFla的Nsp2存在编码12个氨基酸的连续36个核苷酸的缺失，ORF、3存在编码1个氨基酸的3个核苷酸的缺失。这是国内外首次发现PRRSV存在缺失变异现象，研究结果补充和丰富了PRRSV毒株的基因组信息数据，为深入研究该毒株的遗传与变异及其与生物学特性的关系奠定了基础。     

H5亚型禽流感DNA疫苗质粒pCAGGoptiHA5对高致病力禽流感病毒攻击的免疫保护

对H5亚型禽流感DNA疫苗质粒pCAGGoptiHA5的免疫效果进行了研究。pCAGGoptiHA5分别以100和10μg剂量一次或两次免疫3周龄SPF鸡，首次免疫后4周以同样剂量和途径进行第二次免疫，一次免疫后4周、两次免疫后2周分别用100LD50的HPAIV A／Goose／GuangDong／1／96（H5N1）鼻腔途径进行攻击，观察发病与死亡情况，分别于攻毒后第3、5、7天采集喉头及泄殖腔拭子进行病毒分离、滴定检测排毒情况，同时检测免疫后、攻毒前及攻毒后血清HI抗体、NT抗体以及AGP抗体的动态变化。结果，100μg pCAGGoptiHA5一次免疫、100μg pCAGGoptiHA5两次免疫以及10μg pCAGGoptiHA5两次免疫均可对免疫鸡形成100％完全保护（不发病、不致死、不排毒），10μg剂量pCAGGoptiHA5一次免疫可对免疫鸡形成100％的保护（不发病、不致死），结果表明，pCAGGoptiHA5作为疫苗效果良好、成本低廉，有望成为预防H5亚型高致病力禽流感的高效、安全新型基因工程疫苗。     

Comparative Study on Carcass Traits and Meat Quality of Different Month Old of Xinjiang Brown Cattle Steers

In this research,12 heads of 28 to 30 months (low months age group) and 8 heads of 32 to 34 months (high months age group) Xinjiang Brown cattle steers which fed in the same mode of nutrition were chose for the slaughter test,in which the carcass traits,nutrient composition and some edible quality index were measured and comparative analyzed,a research which through comparative analysis the carcass traits and meat quality of nutritional components of different months of Xinjiang Brown cattle steers was carried out.The results indicated that the marbling,carcass grade,fat,meat color of high months age group were better than that of low months age group,the traits of the low months age group on fat color,moisture,protein,cooking loss and shear force value were better than that of high months age group,and there were significant difference on the traits of fat and meat color L^* between the two groups (P<0.05),while the differences of other traits between them were not significant (P>0.05);As high-grade beef,in addition to the traits of meat color and pH value of the chunk roll,tenderloin and striploin,each site of the meat traits on the fat,protein,moisture,cooking loss,shear force value indicators were all better than other parts,and showed the best of nutritional value and edible quality.28 to 34 months of Xinjiang Brown cattle steers could produce high-grade beef,the meat could be used as the raw materials of steak and boiled beef.     

白龙江上游主要灌丛土壤入渗性能分异性

对白龙江上游主要5种灌丛类型进行土壤物理性质和入渗性能试验。结果表明:不同灌丛类型的土壤物理性质和入渗性能都存在着较大的差异,杜鹃(Rhododendron simsii)灌丛土壤蓄水能力和入渗性能均大于其它灌丛类型,臭樱(Maddenia hypoleuca)灌丛土壤蓄水能力和入渗性能最差;不同灌丛类型土壤的初渗率在3.68~44.13 mL/min,稳渗率在1.05~4.71 mL/min,总体上是杜鹃灌丛荚蒾(Viburnum dilatatum)灌丛箭竹(Fargesia spathacea)灌丛中华柳(Salix cathayana)灌丛臭樱灌丛;土壤初渗率和稳渗率与土壤自然含水量、入渗总量和孔隙度呈及显著正相关,和土壤密度呈及显著负相关。     

安徽省发展大径级材森林资源的思考

珍贵大径级材是我国重要的战略资源。探讨了珍贵大径级材在安徽省培育的意义及有利条件等,并提出了相应的政策措施和技术路线。