戊型肝炎病毒颗粒性蛋白疫苗H-2~d限制性Th表位的筛选

戊型肝炎病毒衣壳蛋白重组抗原HEV 239能形成类病毒颗粒,具备演变成多价疫苗的载体的潜力,此文旨在筛选、鉴定其内包含的H-2d限制性Th表位。以50μg HEV 239蛋白与完全弗式佐剂混合后皮下免疫BALB/c鼠,以覆盖HEV 239蛋白全长的15氨基酸肽库体外刺激其脾细胞,用IFN--γELISPOT方法检测其细胞免疫应答,并通过磁珠剔除脾细胞中CD4+T细胞或CD8+T细胞以分析筛选得到的T细胞表位的特性。结果显示:HEV 239中包含优势的T细胞表位P34(HEV PORF2 AA533~AA547,HSKTF FVLPL RGKLS)及数个较弱的T细胞表位,P34对HEV 239免疫的BALB/c鼠脾细胞的刺激效果与HEV 239蛋白相当,剔除实验表明该表位为CD4+T细胞表位,即Th表位

戊肝病毒Th表位肽免疫可增强其载体蛋白的体液免疫应答

戊型肝炎病毒衣壳蛋白内包含一个强H-2d限制性Th表位P34。以该表位肽免疫BALB/c鼠,其脾细胞能够在体外识别重组戊型肝炎病毒衣壳蛋白,剔除实验表明应答细胞几乎完全是CD4+T细胞,证明P34表位肽能有效诱导产生特异性Th细胞。以P34肽初免小鼠,再以包含该表位的重组戊型肝炎病毒抗原(E2)免疫,结果表明,10μg、20μgE2免疫组在免疫后第1周即有部分小鼠产生抗体,到第3周所有小鼠均能够产生抗体;而对照肽P18初免的小鼠,以20μgE2加强免疫亦无法诱导小鼠产生抗体。这表明,Th表位肽P34初免诱导产生的Th细胞能够有效促进小鼠对携带该表位的载体蛋白的体液免疫应答

代谢组学研究进展

代谢组学是功能基因组学和系统生物学研究不可或缺的重要组成部分，是通过考察生物体系受刺激或扰动前后（如将某个特定的基因变异或环境变化后）代谢产物的动态变化，研究生物体系的代谢网络的一种技术。近年来，在药物研发、疾病研究、植物和微生物等领域发展迅速，但在分析技术、方法学和应用等方面急需突破

HPLC 2015

<p>Splat frozen samples of <i>S</i>. <i>feltiae</i> extract (top row) and buffer control (25 mM Tris HCl, pH 8) (bottom row) after warming to – 8°C (Time 0 = left column) and after annealing at– 8°C for 30 min (right column). Scale bar = 100 μm.</p

Liquid chromatography tandem mass spectrometry for determination of catecholamines using alumina-based extraction method

Catecholamines including epinephrine, norepinephrine and dopamine are important neurotransmitters in human species. The levels of catecholamines in biologic sample are utilized to diagnose hypertension, pheochromocytoma and neuroblastoma. Different analytical methods are used for catecholamines, such as high-performance liquid chromatography, capillary electrophoresis, mass spectrometric detection, electrochemical detection, chemiluminescence detection and fluorescence detection. Extraction is needed because of low level and interference in the biologic samples. In this work, a method of liquid chromatography tandem mass spectrometry using multiple reaction monitoring (MRM) in the positive ion mode was developed to analyze catecholamines. Multiple reaction monitoring detection not only improved the sensitivity but also selectivity of the determination. In order to isolate the target compounds from the matrix solutes, an alumina-based extraction method was achieved. Different pH values which influenced the adsorbability of alumina were investigated. The precision and the recovery of this method were satisfactory