人体的免疫功能是维持身体健康的重要生理性功能,在免疫系统的功能受损或低下时,会对人的身体健康产生严重影响.甘草作为一种补益类中药,有补脾益气之功,对人体的免疫功能具有增强作用,在开发成免疫增强新药方面具有独特优势和发展潜力.通过对近期国内外甘草免疫增强作用的相关文献进行收集和整理,从所含化合物的结构特点、分类及药理作用等方面阐述了甘草增强免疫的物质基础,并以甘草的药效成分为基础综述了甘草促进免疫细胞增殖、影响体内细胞因子和抗体水平以及调控相关酶表达等增强免疫的分子机制,以期为免疫增强药物的研发提供新的思路,为甘草免疫作用的进一步研究提供参考.

为了探讨细胞周期抑制因子p19ARF对人二倍体细胞复制性衰老的影响,构建了重组p19ARF真核表达载体,并通过脂质体的介导将p19ARF基因转染到人二倍体成纤维细胞WI-38中过表达,观察其对WI-38细胞衰老的影响.结果发现与对照细胞相比,在p19ARF基因导入后,细胞中p53和p21的表达水平明显上调,细胞传代数减少10～12代,生长速率降低,细胞周期阻滞于G1期,衰老标志物SA-β-gal染色阳性率上升,线粒体膜电位下降,细胞形态呈衰老细胞样变化,这些结果表明p19ARF高表达可促进人二倍体细胞的衰老进程.

Specific anti-hURAT1 rabbit polyclonal antibody was obtained, and both Western-blot and immunohistochemistry showed that hURAT1 was expressed in the human kidney brush border, localized in the apical membrane of the LLC-PK1 cell. hURAT1 protein was a membrane protein located in renal proximal tubule, which could be detected in the apical membrane. The anti-hURAT1 polyclonal antibody could be used for studying the physiological function of hURAT1 and its pathology.

Objective:Ogranic anion transporter 4 (OAT4),which expresses in human kidney, plays a principal role in secreting organic anion metabolized from both endogenous and exogenous compounds.Antibody is an important tool to study hOAT4.This study is aimed to produce mouse anti-hOAT4 polyclonal antibody with high throughput and high specificity by genetic immunization and to detect the localization fo hOAT4 in human kidney.Methods:Intracellular high antigenicity fragment (E278～R345) of hOAT4 was chosen by protein antigenicity prediction software and its cDNA was amplified from human renal total RNA by RT-PCR, which was subsequently cloned into pBQAP-OVA plasmid to construct recombinant plasmid pBQAP-OVA-hOAT4 for genetic immunization. The mice were inoculated with this recombinant plasmid and two other adjuvant plasmids, pCMVi-GMCSF and pCMVi-FlT3L, which helped to enhance the antibody's generation. After 5 weeks, mice were sacrificed to obtain anti-hOAT4 antibody. Antibody titer was measured by ELISA. The antibody was identified by Western blot analysis and immunohistochemistry.Results:The cDNA of hOAT4 intracellular high immunogenic fragment were amplified successfully. Recombinant plasmid pBQAP-OVA-hOAT4 for genetic immunization was confirmed by restriction digestion and sequencing. ELISA assay indicated that mouse anti-hOAT4 antibody has high titer and could recognize a band of 65 kD glycosylated hOAT4 from human renal membrane protein and brush border membrane protein in Western blot.Immunohistochemistry results indicated that hOAT4 located at the brush border membrane of renal proximal tubular cells.Conclusion:Genetic immunization can generate anti-hOAT4 polyclonal antibody with high throughput and specificity.

为观察碱性成纤维细胞生长因子(bFGF)对离体热烫伤后成纤维细胞内钙离子的作用以及丝裂原活化蛋白激酶(MAPK)通路对胞内游离钙的影响,将培养的人成纤维细胞进行热损伤刺激后分成4组:①bFGF处理组(10ng/ml); ②预先加入PD98059(10μmol/L)阻断剂30min,再行bFGF(10ng/ml)刺激;③预先加入SB203580(10μmol/L)阻断剂30min,再行bFGF刺激(10ng/ml); ④同时加入PD98059(10μmol/L)和SB203580(10μmol/L)2种阻断剂30min,再加入bFGF(10ng/ml).应用特异性Ca2+荧光指示剂Fluo-3/AM负载细胞,激光共聚焦显微镜检测细胞内游离钙的浓度.结果显示,受热刺激的成纤维细胞荧光强度较弱.加入bFGF后可促使成纤维细胞中游离Ca2+浓度升高,分别预先加入PD98059和SB203580拮抗剂的成纤维细胞,再加入bFGF,胞内钙离子浓度出现不同的钙振荡现象.同时加入2种阻断剂后胞内钙离子浓度则迅速降低.表明bFGF引起热损伤后的成纤维细胞中游离Ca2+浓度的增加,MAPK信号通路对胞内钙离子有反馈调节的作用.