Objective To observe the effect of LIGHT on synovial macrophage-osteoclast difier entiation in rheumatoid arthritis.Methods Synovial macrophages were collected from 8 synovial tissues har vested from RA patients by digestion with conagenase.The macrophages from each patient were divided into 5 subgroups:group 1 was co-cultured with Macrophage Colony Stimulating Factor(MCSF).group 2 was co cultured with MCSF and LIGHT.group 3 was co-cultured with MCSF and Receptor activator for Nuclear Fac tor kB ligand(RANKL),group 4 was co-cultured with MCSF and LIGHT and RANKL group 5 was co-cul tared with LIGHT.After two weeks of culture.tartrate resistant acid phosphatase(TRAP)and F-actin staining were used to detect the formation of osteoclast on cover slip(CS).Functional evidence of osteoclasts was as sessed by the formation of resorption pits ou dentine slice(DS).Results TRAP and F-actin were both nega tive in group 1 and group 5 and no pit on dentine slice could be observed.In group 2.there were some small round and ovoid osteoclasts with TRAP(+)and F-actin(+).Pits on DS were small and discrete.In group 3. there were many large irregular osteoclasts with TRAP(++)and F-actin(++).The number and volume of pits were both increased.In group 4.there were even more and larger osteoclasts with TRAP(+++)and F-actin (+++).The pits were even larger and became confluent,ie,pits(++++).Conclusion LIGHT can promote RANKL-mediated osteoclastogenesis in RA synovial macrophages.and it can induce osteoclast formation through a mechanism independent of RANKL.

AIM: To investigate the relationships between antiproliferative mechanisms of probucol and protein expressions of signaling molecules ERK1/2, MKP-1, HO-1 and Trx-1 in rat aortic smooth muscle cells (RASMCs) stimulated with ox-LDL. METHODS: The effects of probucol on cell cycle, cell proliferation and the expressions of ERK1/2, MKP-1, HO-1 and Trx-1 in the presence of ox-LDL were observed by means of MTT test, FCM and Western blotting. RESULTS: (1) Probucol significantly inhibited the proliferation of RASMCs stimulated with ox-LDL. A value in 100 μmol/L probucol+35 mg/L ox-LDL group was reduced by 34.9% as compared to ox-LDL group (P<0.01). (2) Probucol protected against ox-LDL-induced RASMCs proliferation through inducing cell growth arrest at G_0/G_1 phase and cell apoptosis. (3) ox-LDL increased the expression of p-ERK1/2 by 34.7% (P<0.01) and decreased MKP-1 by 60.0% (P<0.01), respectively, as compared to control. Probucol attenuated the increase in ox-LDL-stimulated p-ERK1/2 level by 15.7%, but increased MKP-1 expression by 2 times (P<0.01). (4)ox-LDL at concentration of 35 mg/L decreased the intracellular Trx-1 expression by 28.9% (P<0.05), and slightly increased the level of HO-1 expression as compared to control (P<0.05). Probucol enhanced the expression of Trx-1 by 91.6% (P<0.01) and HO-1 by 31.9% (P<0.01), respectively as compared to ox-LDL group. CONCLUSION: Probucol inhibits ox-LDL-stimulated the proliferation of RASMCs through increases in MKP-1/HO-1 expression, suppression of cell cycle progression and induction of cell apoptosis.

Objective To establish a quantitative detection method for Mycobacterium tuberculosis by immunomagnetic capture combined with PCR-ELISA detection system with double internal standards(IMC-PCR-ELISA) .Methods The immunomagnetic (Dynabeads? ) which could specifically capture Mycobacterium tuberculosis were prepared .According to Mtp40 and IS6110 gene sequence of Mycobacterium Tuberculosis ,2 pairs of primers(upstream primer was modified with Biotin at 5′end) ,2 same-length mutant fragments with PCR amplified fragments ,and 3 capture probes(modified with digoxigenin at 3′end) were designed .Myco-bacterium tuberculosis were captured by immunomagnetic ,then detected by PCR-ELISA with double internal standards .Results The IMC-PCR-ELISA method could yield quantitative results in about 4 h with a detection limit at 5 × 103 copies/mL .There was a fine linear relationship between the copies of Mtp40(IS6110)in fact and in the calculation through formula when the concentrations of low internal standards were 30-70 copies/mL and the concentrations of high internal standards were 8 000-12 000 copies/mL (r2 =0 .998) .No nonspecific amplification was observed .Conclusion A rapid and quantitative method for the detection of Myco-bacterium tuberculosis was established successfully .The IMC-PCR-ELISA method was rapid ,sensitive ,secific and quantitative .