ABSTRACT
Objective@#To investigate the methylation status and expression of FOXP3 in CD4+ T cells of patients with chronic hepatitis B (CHB).@*Methods@#Peripheral blood mononuclear cells (PBMCs) from 59 CHB patients and 22 healthy controls (HC) were collected. The percentage of CD4+ CD25+ Foxp3+ Tregs in CD4+ T cells was estimated by flow cytometry. FOXP3 expression was measured by quantitative real time-polymerase chain reaction (RT-qPCR) and Western blotting. The methylation status of FOXP3 was determined by methylation-specific polymerase chain reaction.@*Results@#The percentage of CD4+ CD25+ pFoxp3+ Tregs in CD4+ T cells, FOXP3 mRNA and protein expression levels were significantly higher in patients with CHB than HCs (P<0.05). Meanwhile, the methylation frequency of FOXP3 was significantly lower in CHB patients than HCs (P<0.05). FOXP3 mRNA levels and the percentage of CD4+ CD25+ Foxp3+ Tregs were significantly lower (P<0.05) in patients with gene methylation than those without.@*Conclusions@#Aberrant demethylation of FOXP3 gene existed in CD4+ T cells of CHB, which contributed to an elevation in FOXP3 expression and percentage of CD4+ CD25+ Foxp3+ Tregs. It might provide a new target for prevention and treatment of CHB.
ABSTRACT
iRGD-modified sterically stabilized liposomes loaded doxorubicin (iRGD-SSL-DOX) were prepared and their cellular toxicity and anti-tumor efficacy were evaluated, comparing to doxorubixin loaded sterically stabilized liposomes (SSL-DOX) and RGD modified doxorubixin loaded sterically stabilized liposomes (RGD-SSL-DOX). The iRGD peptide, with both tumor targeting and cell penetrating functions, was conjugated to DSPE-PEG-NHS and DSPE-PEG-iRGD was obtained. DSPE-PEG-RGD was gained in the same way. iRGD-SSL-DOX, RGD-SSL-DOX and SSL-DOX were prepared by ammonium sulfate gradient method. The size and zeta potential of the liposomes were characterized by dynamic laser light scattering. The cellular toxicity study was done on B16 melanoma cell line and the anti-tumor efficacy study was carried on B16 cell line bearing C57BL/6 mice. The results showed that the particle sizes of liposomes were all around 90-100 nm. DOX entrapment efficiency was above 95%. The formulations were with good preparation reproducibility. iRGD-SSL-DOX showed no significant difference in B16 cellular toxicity with SSL-DOX and RGD-SSL-DOX, but the anti-tumor efficacy on B16 melanoma bearing C57BL/6 mice was significantly better than that of SSL-DOX, similar as that of RGD-SSL-DOX. Therefore, iRGD modified liposomes loaded DOX would be a promising drug delivery system for tumor therapy.