小菜蛾腺苷受体基因PxAdoR克隆及杀虫靶标作用

利用蝉花虫草Ophiocordyceps sobolifera代谢产物N6-（2-羟乙基）腺苷（HEA）作用于小菜蛾Plutella xylostella L.获得差异表达的片段,采用RT-PCR和RACE技术从小菜蛾中克隆全长1828 bp的小菜蛾腺苷受体（Plutella xylostella adenosine receptor,PxAdoR）基因,通过生物信息学软件预测和分析蛋白质结构,PxAdoR为分子量49.2 kD,没有信号肽,疏水性不稳定蛋白,具有7次跨膜螺旋结构的一类G蛋白偶联受体（G protein coupled receptor,GPCRs）。利用HEA及哺乳动物腺苷受体A2aR抑制剂SCH58261处理小菜蛾2龄幼虫,结果显示联合使用HEA和SCH58261后36~60 h,可使HEA引起的小菜蛾2龄幼虫的校正死亡率显著降低（42.7%~53.9%,P<0.05）;而联合用药后36、48、60 h小菜蛾2龄幼虫体重增长抑制率由43.3%、38.3%、46.2%显著降低（P<0.05）到13.2%、10.3%、8.6%。本研究表明SCH58261能显著降低HEA引起的小菜蛾2龄幼虫的死亡率和体重增长抑制率,HEA可能是通过PxAdoR而起作用,PxAdoR基因可以作为新的杀虫靶点用于害虫控制研究。     

鸡传染性支气管炎病毒介导的细胞内受体及抗病毒基因的表达

[目的]研究传染性支气管炎病毒对细胞模式识别受体(PRRS)及天然抗病毒基因转录的激活作用.[方法]IBV M41毒株感染DF-1细胞,应用荧光定量PCR测定细胞内受体(MDA5、LGP2、MAVS和NLRC5)和抗病毒相关基因(IRF7,IFN-a/β,NF-κB1)在禽传染性支气管炎感染过程中不同时间点的转录水平表达变化.[结果]在感染的细胞中几种细胞内受体在感染不同时间点均有不同程度的升高.IFN-α、IRF7和NF-κB1的表达在感染后显著升高,而IFN-β在感染36 h内被抑制,48 h被激活.[结论]鸡传染性支气管炎病毒体外感染DF-1细胞后能显著激活细胞内受体及抗病毒基因的表达.     

水貂部分Toll样受体(TLRs)基因的分子克隆、序列分析及组织表达分析

试验旨在研究Toll 样受体(Toll-like receptors,TLRs)在水貂抗病毒免疫中的作用机制,并为抗病育种积累可供选择的基因素材。本试验以雪貂TLRs基因为参考序列设计4对引物对水貂TLR4、TLR6、TLR7及TLR8进行分子克隆,并对所得序列进行生物信息学分析,进一步利用半定量RT-PCR技术分析TLR4和TLR7基因mRNA在不同年龄水貂各组织中的表达情况。结果表明,水貂TLRs与食肉目动物(雪貂、北极熊、大熊猫、海象、海豹、犬、老虎和猫)具有较高的同源性,在系统发育树中距离最近;组织表达分析表明TLR4和TLR7在检测的组织中广泛表达且表达量存在差异,相比成年水貂,仔貂各组织中TLR4或TLR7基因表达量更高。     

核受体PPARs与动脉硬化

过氧化物酶体增殖激活受体 (PPARs)是配体激活转录因子 ,存在 3种亚型 ,在多种细胞组织内广泛分布。动脉硬化的特征性变化之一是巨噬细胞源和平滑肌源泡沫细胞在内皮下大量聚集 ,血脂代谢障碍和免疫因素是及其重要的病理生理变化。近年来研究发现 ,PPARα、PPARγ也在血管壁的多种细胞中表达 ,在其配体作用下 ,抑制某些致病基因的表达 ,对动脉粥样硬化斑块的形成和发展具有重要的调节作用     

Transition period-related changes in the abundance of the mRNAs of adiponectin and its receptors,of visfatin,and of fatty acid binding receptors in adipose tissue of high-yielding dairy cows

Adipose tissue expresses adipokines, which are involved in regulation of energy expenditure, lipid metabolism, and insulin sensitivity. To adapt for the transition from pregnancy to lactation, particularly in high-yielding dairy cows, adipokines, their receptors, and particular G-protein coupled receptors (GPRs) are of potential importance. Signaling by GPR 41 stimulates leptin release via activation by short-chain fatty acids; GPR 43/109A inhibits lipolysis, and GPR 109A thereby mediates the lipid-lowering effects of nicotinic acid and β–hydroxybutyrate. The aim of this study was to compare the mRNA expression of adiponectin and visfatin, adiponectin receptors 1 and 2 (AdipoR1/2), leptin receptor (obRb), insulin receptor as of the aforementioned GPRs during the transition period in high-yielding dairy cows. Biopsies from subcutaneous fat and blood samples were obtained from 10 dairy cows 1 week before and 3 weeks after calving. For AdipoR1 and AdipoR2 mRNA abundance as well as for leptin concentrations in plasma, a reduction (P ≤ .05) was observed postpartum; for visfatin and putative GPR 109A mRNA abundance in adipose tissue, there was a trend (P < .1) for analogous changes. In contrast, the mRNA content of obRb and GPR 41 in adipose tissue was higher (P ≤ .05) in samples from early lactation than in those from late gestation. Our results indicate decreasing adiponectin sensitivity in adipose tissue after calving, which might be involved in the reduced insulin sensitivity of adipose tissue during early lactation. In addition, visfatin, GPR 41, and GPR 109A may further modulate insulin sensitivity.     

Immunomodulatory effects of β-glucans on porcine alveolar macrophages and bone marrow haematopoietic cell-derived dendritic cells

The immunopharmacological activities of β-glucans with a backbone of β-1,3/β-1,6-linkages associated with anti-tumor, anti-viral, bacterial and fungal infections have been well documented. Dectin-1, a specific pattern recognition receptor for β-1,3/β-1,6-glucans, is expressed mainly on phagocytes, especially macrophages and dendritic cells (DCs). In this study, the encoding nucleotide for the carbohydrate-recognition domain (CRD) of porcine dectin-1 was sequenced for the first time, and the immunomodulatory functions of a synthetic particulate β-glucan (p-β-glucan) were examined. Results showed that p-β-glucan significantly enhanced cell activity and phagocytosis in porcine alveolar macrophages (AMs), immature DCs (imDCs) and mature DCs (mDCs), in a similar way to zymosan. Zymosan enhanced dectin-1/TLR2/TLR4 expression and TNF-α/IL-10 production in all of three types of cell, whereas p-β-glucan increased dectin-1/TLR4 and TNF-α/IL-12 production in AMs but inhibited IL-10 in mDCs. These results indicate that the complex collaborating interactions between dectin-1 and TLRs in the recognition of β-1,3/β-1,6-glucans with different structural features may direct different cellular responses.     

鲭鱼鱼肉中组胺菌的分离及其理化性质分析

利用组氨酸肉汤培养液,对鲭鱼鱼肉中的组胺生成菌进行筛选,然后通过生物化学和形态学实验,进而对组胺生成菌进行分析。结果表明：分离得到2株组胺菌,菌种I在5～25℃的培养温度范围内,随温度升高,组胺生成量逐渐增加,在25℃组胺生成量最高,约2700mg／kg。菌种Ⅱ在20℃组胺生成量最高,约为2000mg／kg。依据2种组胺菌的特征和生理生化特征,分离菌可能分别属于摩尔摩根氏菌、嗜水性气单胞菌属。     

市售鱼类及其加工品中组胺含量的调查

2006年3月对市售鲭鱼20尾、秋刀鱼30尾、金枪鱼6块等进行组胺含量分析,结果表明,20尾鲭鱼中有1尾组胺含量为20mg／kg;30尾秋刀鱼肉中有1尾组胺含量为16mg／kg,另一尾为10mg／kg;6块金枪鱼样品及其他鱼类样品未能检测到组胺。鲭鱼20尾中1尾检出酪胺23mg／kg,尸胺为36mg／kg,腐胺及亚精胺未检出;秋刀鱼30尾中2尾检出10mg／kg和15mg／kg的酪胺,1尾栓出52mg／kg腐胺,2尾检出35mg／kg和55mg／kg尸胺。     

Leucettamols, Bifunctionalized Marine Sphingoids, Act as Modulators of TRPA1 and TRPM8 Channels

Leucettamols, bifunctionalized sphingoid-like compounds obtained from a marine sponge Leucetta sp., act as non-electrophilic activators of the TRPA1 channel and potent inhibitors of the icilin-mediated activation of the TRPM8 channel, while they are inactive on CB₁, CB₂ and TRPV1 receptors. Leucettamols represent the first compounds of marine origin to target TRPA1 and the first class of natural products to inhibit TRPM8 channels. The preparation of a small series of semi-synthetic derivatives revealed interesting details on the structure-activity relationships within this new chemotype of simple acyclic TRP modulators.