ABSTRACT
Objective: To investigate the differential expression of serum soluble TNF-related apoptosis-inducing ligand (sTRAIL) between ankylosing spondylitis (AS) and rheumatoid arthritis patients (RA) and to discuss its clinical significance. Methods: Sixty AS patients, including 38 HLA-B27-positive ones and 22 HLA-B27-negative ones, 20 rheumatoid arthritis (RA) patients and 30 healthy individuals were included in the present study. The AS patients were divided into active group and inactive group based on bath ankylosing spondylitis disease activity index (BASDAI). The concentrations of serum sTRAIL were measured by ELISA in all groups. Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were detected automatically by ESR automatic analyzer and specific protein analyzer. Results: The serum sTRAIL concentration was significantly higher in AS patients (both HLA-B27-positive and -negative AS patients) than in RA patients and healthy controls (P<0.01); no significant difference was found between HLA-B27-positive and -negative AS patients. Serum sTRAIL concentration was significantly higher in active AS group than in inactive AS group (P<0.01). Serum sTRAIL and CRP concentrations were correlated with each other in HLA-B27 positive AS patients (r = 0.609, P = 0.000), but not in HLA-B27-negative ones. Serum sTRAIL concentration was not correlated with ESR in AS patients. Conclusion: Serum sTRAIL is obviously up-regulated in AS patients, which is not associated with the status of HLA-B27. However, the association between sTRAIL and CRP is influenced by the status of HLA-B27.
ABSTRACT
Objective To establish a specific fluorescence quantitative method for determining the mRNA expression of Toll-like receptor3(TLR3)in peripheral blood mononuclear cells(PBMCs).Methods Using the Beacon Designer 2.1 software,specific primers and Taqman-MGB probe were designed.The plasmid pMD18-T-TLR3 was constructed as calibrator and the amplified fragment was obtained by reverse- transcript-PCR(RT-PCR).RNA quantification based on cycle threshold values(Ct)was used to establish the standard curve.According to which,the TLR3 mRNA levels in 30 normal individuals,20 patients with primary biliary cirrhosis(PBC)and 20 ones with chronic liver cirrhosis induced by HBV were calculated automatically by software after the fluorescence of PCR product was detected continuously during amplification.Results The linear detection range of the assay for TLR3 gene and ?-actin was 10~2-10~8(r= -0.9974)and 10~3～10~8(r=-0.9984),respectively.The coefficient of variation of both intra-and inter- assay reproducibility for high concentration sample were 6.7% and 8.7%,respectively,and those for low concentration sample were 12.3% and 14.0%.The TLR3 mRNA expression level ranges from 3.46?10~2- 4.51?10~3 copies/?g RNA,4.92?10~2-1.42?10~4 copies/?g RNA and 2.58?10~2-7.17?10~3 copies/?g RNA for 30 healthy individuals,20 PBC patients and 20 ones with chronic liver cirrhosis induced by HBV, respectively.Conclusion We have successfully set up a FQ-RT-PCR method for detecting TLR3 mRNA, which may be used as an excellent tool for the clinic and basic study on the expression of TLR3 gene.