Construction of HIV-1 broadly neutralizing antibody gene eukaryotic expression vector and detection of its biological function / 中华实验和临床病毒学杂志

Objective@#To construct a recombinant double gene co-expressing plasmid of HIV-1 broadly neutralizing antibody and to detect its expression in 293T cell line.@*Methods@#HIV-1 neutralizing antibody 2G12 variable region of light chain (VL) and heavy chain (VH) was synthesized, and ligated with vectors containing human IgG constant regions of light and heavy chain to construct a complete 2G12 light and heavy Chain. The VL and VH of 2G12 and IRES were cloned into eukaryotic expression vector pVR by PCR amplification and then the recombinant plasmid was transfected into 293T cells. Expression of the antibody in cell supernatant was detected by ELISA. Binding and neutralizing activity of the cell supernatant were tested by ELISA and micro-neutralization assays.@*Results@#The recombinant double gene eukaryotic expression vector which can express human IgG was constructed successfully. The expression level of the supernatant was 6.43 μg/ml, and the antibody retained the binding and neutralizing activity.@*Conclusions@#The constructed vector can express the antibody with binding and neutralizing activity, this study provides a good platform for the expression of human HIV neutralizing antibody in the eukaryotic expression vector.

Effect of Zhengan Xifeng decoction on Proteomics of peripheral blood mononuclear cell in intracerebral hemorrhage patients with liver yang forming wind syndrome / 中华中医药杂志

Objective:With proteomic approach,to find the differentially expressed proteins spots in the peripheral blood mononuclear cells(PBMCs) of healthy people and intracerebral hemorrhage patients with liver yang forming wind syndrome(LYFWS) treated by Zhenganxifeng decoction(ZGXFD) to investigate the possible proteins of LYFWS and the cure mechanism of ZGXFD.Methods: 10 intracerebral hemorrhage patients with LYFWS were treated by ZGXFD for 7-10 days.10 healthy people were selected taken as healthy controls.10ml venous blood was drawed from every subjects and PBMCs were isolated from blood by using lymphocytes separation medium.The total proteins was extracted from PBMCs.The total proteins from either intracerebral hemorrhage patients or healthy controls were prepared by means of immobilized pH gradient based on two-dimensional gel electrophoresis.After Coomassie brilliant blue G250 staining,gel-image analysis was performed by PDQuest.The differentially expressed proteins spots were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry(MALD I-TOF-MS).Results: Gel-image analysis revealed that there were 22 proteins spots expressed differentially.20 proteins spots were expressed differentially in the intracerebral hemorrhage patients with LYFWS as compared with healthy control,of which 8 proteins were down-regulated,7 proteins were up-regulated and 5 proteins were lost.After treated by ZGXFD,4 proteins were down-regulated,5 proteins were up-regulated and 2 proteins were lost,9 proteins weren’t expressed di erentially as compared with healthy control.8 of 20 di erentially expressed proteins spots in the intracerebral hemorrhage patients with LYFWS identi ed by MALDI-TOF-MS as compared with healthy control.These proteins are related to cell metabolism,signal transduction and so forth,of which 6 proteins were down-regulated,1 proteins were up-regulated and 1 proteins were lost.After treated by ZGXFD,2 proteins were down-regulated,1 proteins were lost,5 proteins weren’t expressed di erentially as compared with healthy control.Conclusion: 2-DE pro les of intracerebral hemorrhage patients with LYFWS in PBMCs had been established.The proteins of intracerebral hemorrhage patients with LYFWS had been identi ed,which were related to cell metabolism,signal transduction and so forth.This research presented that ZGXFD could adjust multiple di erentially expressed proteins of intracerebral hemorrhage patients with LYFWS.It was also indicated that the cure mechanism of ZGXFD was to regulate multitarget proteins.

Proteomics in Cancer Research / 生物化学与生物物理进展

The proteomics definition, investigation method and its a pplication in cancer research were simply introduced. Proteomic research is to r eveal the function of genes from an integrated, kinetic and quantitative view at the global protein level, which is an important component of post-genome proje ct. Cancer is a kind of complex disease involved by multi-genes. Proteomic rese arch will be helpful to discover the mechanism of cancer development, to find sp ecial malignant tumor markers and targets of drug treatment.