Expression and purification of rabies virus glycoprotein and analysis of its specific binding capacity to memory B cells / 生物工程学报

We aimed to express and purify three rabies virus glycoproteins with different tags and sizes. After analyzing their binding function, we wish to obtain a rabies virus glycoprotein with higher affinity and ability to specifically bind memory B cells. Experiments were carried out to express full length, as well as the ectodomain RVG by gene engineering method. Combined with the antibody of CD19 and CD27, the candidate protein labeling with fluorescence was used to analyze its binding function. Flow cytometry was used to detect the anti-rabies virus specific memory B cells in PBMCs, and confirm the binding ability between the candidate proteins and anti-rabies virus-specific memory B cells. We successfully constructed three expression vectors pGEX-5X-1-RVG, pET28a-RVG and pET30a-G. Three glycoproteins GST-RVG, His-RVG and His-G were obtained by optimized expression and purification conditions. The antigen specificity of purified GST-RVG, His-RVG and His-G were identified by Western blotting and ELISA. The affinity of these three purified glycoproteins to anti-rabies virus antibody were detected by competitive ELISA. Anti-rabies virus specific memory B cells in positive PBMCs gained from people who had ever been injected with the vaccine can be detected by flow cytometry. Thus, we got a recombinant rabies virus glycoprotein that had high-affinity and could sort antigen specific memory B cells.

Clinical significance of visfatin and cTnI in patients with unstable angina pcetoris after coronary stent implantation / 中国医师杂志

Objective To investigate the changes and clinical significance of serum visfatin and cardiac troponin I (cTnI) level in patients with unstable angina pectoris (UAP) after percutaneous coronary intervention (PCI).Methods The 108 patients (aged 51 ～80 years) with UAP and 52 healthy subjects (aged 48 ～ 75) were enrolled.Before and after PCI at the different time,the serum levels of visfatin and cTnI were analyzed.During three years followed-up,the major adverse cardiac events (MACE) were assessed.MACE defined as cardiac death,myocardial infarction,target lesion revascularization anginal status.Results Twenty four hours later after PCI,compared with the level before PCI,the serum visfatin and cTnI levels of the UAP group were significantly increased (P =0.01) ; the serum Visfatin levels of the UAP group before PCI and 30 days later after PCI had no significant difference(P =0.12) ; Thirty days later after PCI,the serum cTnI levels of the UAP group were lower than them before PCI (P =0.02) ; high level visfatin and cTnI were independent predictors of MACE in UAP after PCI (OR =3.27,2.85) (P ＜0.05).The serum cTnI and visfatin levels were significantly positive correlation (r =0.523,P =0.01).Conclusions The serum visfatin and cTnI levels in patients with UAP after PCI increased transitorily,and they had certain value in predicting MACE.