AIM: To construct a recombinant adenovirus expression vector containing CTLA4Ig gene.METHODS: The CTLA4Ig gene derived from the plasmid PCDNA3.0/CTLA4Ig by using polymerase chain reaction (PCR) was inserted into the backward position of cytomegalovirus (CMV) immediate early promoter of the shuttle plasmid (pAdTrack-CMV). After being identified by endonuclease, PCR and sequencing, the recombinant shuttle plasmid pAdTrack-CTLA4Ig was co-transformed into E.coli. BJ5183 cells with the adeoviral backbone plasmid pAdEasyl-1 to obtain the homologous recombination. The adenovirus was generated in 293 cells. A series methods such as PCR and fluorescence microscope was employed to identify the generated recombinant adenovirus. RESULTS: Recombinant CTLA4Ig adenoviruses were constructed and the titer of virus was generally up to 1.65?10 12 phaque forming units per liter (PFU/L). CONCLUSION: Success in constructing recombinant pAdTrack-CTLA4Ig will be the base of the further research on its expression in the mammalian cells, and be potenially used in the prevention of transplant rejection and autoimmunity diseases.

Objective To investigate the autophagy and the expression of its related genes in the peripheral blood mononuclear cells (PBMCs) of active systemic lupus erythematosus (SLE) patients.Methods Patients with newly onset or recently-diagnosed SLE (n=20) were enrolled.RA patients (n=10) and healthy blood donors (n=10) were used as controls.PBMCs from all subjects were immediately isolated by Ficoll-Hypaque density gradient centrifugation.And then monocytes were removed by wall sticking method.The morphology was observed using transmission electron microscopy (TEM).Messenger RNA (mRNA) expression of Beclin 1 and microtubule-associated protein 1-light chain 3 (MAPLC3) were determined by reverse transcription polymerase chain reaction (RT-PCR) and real-time quantitative RT-PCR respectively.Results TEM showed autophagic phenomenon in PBMCs from active SLE.On the mRNA level,expression of Beclin 1 and LC3 was significantly increased in fresh isolated SLE cells as compared with RA or healthy donor's PBMCs.Conclusion Based on these results,we can conclude that autophagy occurs in active SLE and the expression of its related genes is significantly higher in active SLE than in RA or normal controls.The enhanced autophagy may indicate its role in the pathogenesis of SLE.

AIM: To investigate the effect of IL-1 gene polymorphism on the expression of IL-1? mRNA in patients with rheumatoid arthritis. METHODS: The method of FQ-RT-PCR was used to detect the expression of IL-1? mRNA in peripheral blood mononuclear cells separated from rheumatoid arthritis patients with different IL-1? genotype. RESULTS: The expression levels of IL-1? mRNA in 20 patients who carried IL-1? 2*2 genotype were higher than patients who carried no 2*2 genotype and normal subjects. Significant difference existed among three groups. CONCLUSION: IL-1? gene polymorphism influences the transcription of IL-1?. [

Atherosclerosis is a chronic systemic inflammatory disease caused by the interaction of environment and genetic factors. Epigenetic modification is a bridge between environmental factors and genetic factors. DNA methylation is an important regulatory mode of epigenetic modification, which can regulate gene expression at the pre-transcriptional level. Studies have shown that DNA methylation plays an important role in the occurrence and development process of atherosclerosis. Therefore, atherosclerosis-related DNA demethylation, especially carotid atherosclerosis-related DNA demethylation, can be achieved by natural or synthetic DNA demethylases, thereby achieving the purpose of preventing or treating atherosclerotic cerebrovascular disease. This article reviews DNA methylation and its relationship with atherosclerosis and DNA demethylation therapy.