ABSTRACT
BACKGROUND: In this study, we detected plasma urokinase plasminogen activator (uPA) and soluble urokinase-type plasminogen activator receptor (uPAR) levels in Chinese lupus nephritis patients from a large cohort. The associations between plasma uPA and soluble uPAR and clinico-pathological characteristics were further analyzed. METHODS: The levels of plasma uPA and soluble uPAR were detected by ELISA in 202 patients with active lupus nephritis, 17 systemic lupus erythematosus (SLE) patients without renal involvement and 21 normal controls. RESULTS: There were no significant differences in the levels of the average plasma uPA among the lupus nephritis group, non-renal SLE group and normal control group (p = 0.129). The plasma-soluble uPAR level in the lupus nephritis group was significantly higher than that in the non-renal involvement SLE group (p = 0.004) and that in normal controls (p < 0.001). The plasma uPAR levels were positively associated with SLEDAI scores (r = 0.215, p = 0.007). In renal pathological data, there was significant difference of plasma-soluble uPAR levels among various pathological classes, which was the highest in the class IV group (p = 0.012). The level of plasma-soluble uPAR was found to be a risk factor for long-term renal outcomes in lupus nephritis by univariate survival analysis (p = 0.013, HR = 6.326, 95% CI: 1.466-27.298). CONCLUSIONS: Our study showed that the significantly increased plasma levels of soluble uPAR could be found in active lupus nephritis, and they were associated with some clinico-pathological features. Its involvement in the pathogenesis of lupus nephritis warrants further study.
Subject(s)
Lupus Nephritis/blood , Lupus Nephritis/pathology , Receptors, Urokinase Plasminogen Activator/blood , Urokinase-Type Plasminogen Activator/blood , Adult , Biopsy/methods , China/epidemiology , Cohort Studies , Enzyme-Linked Immunosorbent Assay , Female , Glomerular Filtration Rate , Humans , Lupus Nephritis/epidemiology , Lupus Nephritis/physiopathology , Male , Risk FactorsABSTRACT
Hepatitis B virus S protein (HBs) plays an important role in hepatocellular carcinoma progression. However, to date, no direct and effective methods exist to research the function of HBs. Here, we combined the technology of RNA interference with recombinant adenovirus, constructed a recombinant adenovirus-expressing small hairpin RNA of HBs, and infected HepG2.2.15 cells. Then, reverse transcription-polymerase chain reaction (RT-PCR), quantitative real-time PCR, enzyme-linked immunosorbent assay, and Western blot analysis were performed to verify the interference effects. As a result, a recombinant adenovirus was successfully constructed and effectively packaged in AD293 cells, and it significantly inhibited HBs mRNA and protein expression in vitro. Our study may provide a novel tool to study HBs function.
Subject(s)
Gene Expression Regulation, Viral , Hepatitis B virus/genetics , Protein S/genetics , RNA, Small Interfering/genetics , Adenoviridae/genetics , Hep G2 Cells , Hepatitis B virus/pathogenicity , Humans , Protein S/isolation & purification , RNA, Messenger/biosynthesisABSTRACT
HepG2.2.15 cell is a widely used cell model for studying HBV (hepatitis B virus) in vitro. In these cells, the HBV genome is integrated in several sites of HepG2 cellular DNA. These multiple copies may have some influence on the cellular processes. We constructed a new plasmid, pSEH-Flag-HBV, and transfected it into HepG2 cells, and then screened it with hygromycin. We then used ELISA, PCR, and RT-PCR to detect the expression of HBV in these cell lines. A cell line that stably expressed hepatitis B e antigen (HBeAg) and hepatitis B surface antigen (HBsAg) was established. Using Southern blotting analysis, we found that the HBV genome was integrated as a single copy in the cellular DNA. This cell line will be a useful alternative model for HBV studies.
