抗禽流感病毒多表位DNA疫苗的构建及其免疫效力研究

多表位DNA疫苗是建立在常规DNA疫苗基础上的一种新型疫苗。它是用表位作免疫原,这样就比较容易在一个表达载体上克隆病原体的多个抗原基因中具有免疫活性的部分。本试验以H5N1亚型禽流感病毒的HA和NP基因及其表位为基础构建了4个重组质粒：1 pIRES/HA（表达全长的HA基因）;2 pIRES/tHA（只表达HA基因的主要抗原表位区）;3 pIRES/tHANpep（融合表达HA基因的抗原表位区和NP基因的3个CTL表位）;4 pIRES/tHANpep-IFN-γ（用鸡的IFN-γ基因取代质粒pIRES/tHANpep中的neo基因）。分别用这4个重组质粒和空载体质粒pIRES1neo肌注免疫30日龄SPF鸡。免疫3次,间隔为2周,每次每只鸡的剂量为200μg。第3次免疫后两周以高致病性禽流感病毒H5N1强毒攻击,免疫及攻毒前后均采血检测HI抗体效价和外周血CD4⁺、CD8⁺T细胞的变化。结果发现,攻毒前各质粒免疫组均检测不到HI抗体,攻毒后1周存活鸡HI抗体效价迅速升高到64～256。流式细胞仪检测显示外周血CD4⁺、CD8⁺T细胞在疫苗免疫后都有不同程度的升高。空载体质粒对照组鸡（10只）在攻毒后3～8 d内全部死亡,其他各重组质粒免疫组鸡都获得了部分保护,保护率分别是：pIRES/HA组为545%（6/11）,pIRES/tHA组为30%（3/10）,pIRES/tHANPep组为36.3%（4/11）, pIRES/tHANPepIFNγ组为50%（5/10）。这些结果表明我们构建的多表位DNA疫苗能够诱导机体产生特异性免疫应答,并在同型禽流感强毒攻击时对鸡只提供了一定的保护。     

High‐fat diet‐induced dysbiosis mediates MCP‐1/CCR2 axis‐dependent M2 macrophage polarization and promotes intestinal adenoma‐adenocarcinoma sequence

High‐fat diet (HFD) is a well‐known risk factor for gut microbiota dysbiosis and colorectal cancer (CRC). However, evidence relating HFD, gut microbiota and carcinogenesis is limited. Our study aimed to demonstrate that HFD‐induced gut dysbiosis promoted intestinal adenoma‐adenocarcinoma sequence. In clinical study, we found that HFD increased the incidence of advanced colorectal neoplasia (AN). The expression of monocyte chemoattractant protein 1 (MCP‐1), CC chemokine receptor 2 (CCR2) and CD163 in CRC patients with HFD was significantly higher than that in CRC patients with normal diet. When it comes to the Apc^min/+ mice, HFD consumption could induce gut dysbiosis and promote intestinal carcinogenesis, accompanying with activation of MCP‐1/CCR2 axis that recruited and polarized M2 tumour‐associated macrophages. Interestingly, transfer of faecal microbiota from HFD‐fed mice to another batch of Apc^min/+ mice in the absence of HFD could also enhance carcinogenesis without significant body weight gain and induced MCP‐1/CCR2 axis activation. HFD‐induced dysbiosis could also be transmitted. Meanwhile, antibiotics cocktail treatment was sufficient to inhibit HFD‐induced carcinogenesis, indicating the vital role of dysbiosis in cancer development. Conclusively, these data indicated that HFD‐induced dysbiosis accelerated intestinal adenoma‐adenocarcinoma sequence through activation of MCP‐1/CCR2 axis, which would provide new insight into better understanding of the mechanisms and prevention for HFD‐related CRC.     

高原环境与肠道微环境间的关系研究

为了便于进一步认识和深入研究高原环境与机体肠道微环境间的关系,本文就高原环境的特点,肠道微环境的病理学改变,分类,治疗和预后等相关研究做一简要综述。     

Cloning, sequencing, and expression of the gene encoding extracellular alpha-amylase from Pyrococcus furiosus and biochemical characterization of the recombinant enzyme.

The gene encoding the hyperthermophilic extracellular alpha-amylase from Pyrococcus furiosus was cloned by activity screening in Escherichia coli. The gene encoded a single 460-residue polypeptide chain. The polypeptide contained a 26-residue signal peptide, indicating that this Pyrococcus alpha-amylase was an extracellular enzyme. Unlike the P. furiosus intracellular alpha-amylase, this extracellular enzyme showed 45 to 56% similarity and 20 to 35% identity to other amylolytic enzymes of the alpha-amylase family and contained the four consensus regions characteristic of that enzyme family. The recombinant protein was a homodimer with a molecular weight of 100,000, as estimated by gel filtration. Both the dimer and monomer retained starch-degrading activity after extensive denaturation and migration on sodium dodecyl sulfate-polyacrylamide gels. The P. furiosus alpha-amylase was a liquefying enzyme with a specific activity of 3,900 U mg-1 at 98 degrees C. It was optimally active at 100 degrees C and pH 5.5 to 6.0 and did not require Ca2+ for activity or thermostability. With a half-life of 13 h at 98 degrees C, the P. furiosus enzyme was significantly more thermostable than the commercially available Bacillus licheniformis alpha-amylase (Taka-therm).     

水稻叶绿素合成缺陷突变体及其生物学研究进展

叶绿素是植物叶绿体内参与光合作用的重要色素,叶绿素合成缺陷突变是一类明显的性状突变,其在理论研究和实际应用方面均具有重要的意义。本文介绍了国内外在水稻叶绿素合成缺陷突变体的发掘、作用机理及其基因定位等方面的研究进展。     

天名精倍半萜内酯化合物

从菊科天名精全草中分得天名精内酯酮,特勒内酯,11（13）－二氢特勒内酯和异埃瓦内酯。其中11（13）－二氢特勒内酯和异埃瓦内酯为新的天然产物。     

我国新分离虫媒病毒的初步鉴定

1990－1994年,从新疆地区的蚊、蜱和病人血清分离了多株病毒,为了明确这些病毒的分类地位,对其中的20株病毒进行了组织培养细胞感染实验和血清学检验,对部分毒株做了动物接种实验和理化性质鉴定。结果显示：20株病毒均可使BHK－21细胞病变（1－3天）,主要表现为细胞圆宿、聚集,融合,破碎,脱落等;致Vero细胞病变为2－4天;15株病毒致C6／36细胞病变（2－4天）,5株病毒对C6／36细胞连续观察7天未见细胞病变。11株病毒对乳鼠2－4天致死,对成年鼠2－5天致死。选取6株病毒进行理化性质鉴定,4株病毒（90260、91002、91004和91028）对5－氟脱氧尿苷耐受,对乙醚和酸敏感,提示为有膜RNA病毒;一株病毒（90265）对5－氟脱氧尿苷、乙醚和酸均敏感,提示为有膜DNA病毒;另一株病毒（9059）对5－氟脱氧尿苷耐受,对乙醚和酸也耐受,提示可能为无膜RNA肠道病毒。20株病毒中,17株病毒与甲病毒、乙型脑炎病毒和布尼亚病毒的特异性免疫腹水不反应,提示这些病毒中可能不存在甲病毒、黄病毒和布尼亚病毒;3株病毒（90260、91002和91004）只与甲病毒的特异性免疫腹水反应,与乙型脑炎病毒和布尼亚病毒的不反应,提示这三株病毒为甲病毒。     

阿拉善干旱荒漠区不同植被类型土壤种子库研究

[1]兰州大学草地农业科技学院,兰州730020 [2]甘肃草原生态研究所,兰州730020 [3]内蒙古阿拉善盟草原站,巴彦浩特750360