ObjectiveTo study B-cell activating factor(BAFF)/a proliferation inducing ligand (APRIL)protein and mRNA expression levels in patients with primary Sj(o)gren's syndrome(pSS)and its contribution to the pathogenesis of pSS.Methotis The levels of serum BAFF and APRIL were tested by ELISA method in 32 pSS patients and compared with 23 healthy control sabjects.BAFF/APRIL protein levels of labial salivary glands were examined in 17 pSS patients by immunohistochemical staining and compared with 8 healthy controls.BAFF/APRIL mRNA expression relative levels to GAPDH of PBMC were also detected in 18 pSS patients by real time RT-PCR and compared with 20 healthy controls.The relationship between BAFF/APRIL levels and several clinical manifestations in pSS patients were analyzed.Results The median level of serum BAFF in pSS were markedly higher than that in healthy controls[(7.4±3.9)ng/ml vs(3.7±1.1)ng/ml,P＜0.01].Serum APRIL levels were also significantly elevated compared to healthy controls[(26±15)ng/ml vs(16±15)ng/ml,P=0.039].The higher levels of BAFF were associated with higher levels of IgG,ESR.total globulin and the titer of ANA(P＜0.05).There was a positive correlation between serum APRIL levels and ESR and labial salivary gland lymphocyte focus scores(P＜0.05).The two cytokines levels inversely correlated with the count of peripheral white blood cells and platelets in pSS(P＜0.05).When comparing serum Ievels of APRIL and BAFF in patients with pSS,a positive correlation could be found(r=0.534,P=0.002).The BAFF/APRIL protein levels were higher in infiltrating lymphocytes of labial salivary glands from pSS patients than from the controls(P＜0.05).The BAFF mRNA[(0.023±0.024)vs(0.245±0.188),P＜0.01]and APRIL mRNA[(0.047±0.035)vs(0.130±0.097),P=0.002]levels of PBMC were significantly lower in pSS patients than in the normal controls.Conclusion The BAFF/APRIL levels of serum and labial salivary glands are significantly elevated in patients with pSS and correlate with clinical parameters.These indicate that BAFF and APRIL may be involved in the pathogenesis of pSS.The low BAFF mRNA and APRIL mRNA levels of PBMC in pSS may reveal that the production and regulation of this system is complicated.

Objective To explore the association between serum cartilage oligomeric matrix protein (COMe) levels and joint destruction in rheumatoid arthritis (RA) and inflammatory arthritis patients. Methods Serum level of COMP was measured with ELISA in 154 arthritis patients.The results of serum COMe were compared with clinical parameters and the modified SHARe scores after 2 years in RA patients. Results Serum COMP level of RA patients was higher than that of normal controls and patients with other forms of arthritis in which only synovial membrane was involved (P＜0.05).We found a significant difference between sCOMP in osteoarthritis (OA) and psoriatic arthritis (PSA) patients with normal controls ,but not with other forms of arthritis.There was no difference of sCOMP between RA,OA or PSA patients.We found a positive correlation between baseline sCOMP and deterioration of modified SHARP scores after 2 years (P＜ 0.001,r=0.848),but did not find correlation between the baseline sCOMP and the titer of anti-CCP,RF, grade of joint function and modified SHARP scores at baseline.We also found a significant correlation between sCOMP and ESR or CRP levels,duration of morning stiffness,joint swelling scores and tenderness scores re spoctively (P＜0.05).Conclusion Serum COMP level is elevated in arthritis patients whose joint cartilage is also involved,particularly in patients with RA.The results of this study indicate that serum COMe may be used as a diagnostic and prognostic marker for cartilage destruction in established RA patients.

Objective To study the association of serum cartilage oligomeric matrix protein (COMP)with disease activity and early joint destruction of rheumatoid arthritis (RA). Methods The serum levels of COMP were measured with ELISA in 94 patients with RA and 40 controls. The serum level of erythrocyte sedimentation rate (ESR), rheumatoid factor (RF), anti-cyclic citrullinated peptide antibodies (ACCP) and hand X-ray were examined at the same time. Results Significantly increased serum level of COMP was found in RA patients (11.3±5.2) U/L as compared to that in healthy controls (9.2±1.7) U/L (P=0.017).Serum level of COMP was higher in 64 active patients (14±6) U/L than that in 30 inactive disease (9±4) U/L(P=0.005). COMP level was positively correlated with the number of the affected joints, X-ray stage, CRP and ESR level (P<0.05); but had no correlation with age, disease course, grade of joint function, RF and ACCP levels. Thirty patients were followed for two years and their radiographic changes were evaluated at the baseline and the end of this study. Sixteen of 18 patients with high concentration serum COMP level had radi-ologic progression, but only 5 of 12 patients with no increase of serum COMP had radiologic progression. A significant difference (P=0.013) was founded in the two groups. Conclusion The present data suggests that the level of COMP is high in patients with RA. High serum levels of COMP indicate high disease activity and early progressive of bone destruction in RA patients. We can us COMP as a laboratory marker of RA.

Objective To study the effects of p27kip1 gene on the apoptosis in esophageal carcinoma cell lines EC 9706. Methods A replication deficient adenovirus vector encoding p27kip1(ad p27kip1) was constructed and human esophageal carcinoma cell lines EC 9706 were transduced in vitro. The cell apoptosis were determined by flow cytometry, TUNEL technique and DNA fragmentation analysis. Results Ad p27kip1 containing the p27kip1 sequence was successfully constructed and the virus titer was 1.24?10 12 CFU/ml. The transduction efficiency was 100% when multplicity of infection≥50. FCM analysis revealed a sub G 1 cell peak in ad p27kip1 transduced esophageal carcinoma cell lines. Agarose electrophoresis showed marked ladder. The difference of apoptotic index between the ad p27kip1 group and the control group was statistically significant(37.3?3.4 vs. 1.3?0.2, P

Objective To investigate the effects of inhibiting ubiquitin proteasome pathway(UPP) on proliferation of gastric carcinoma cells and the possible mechanism was discussed. Methods The gastric carcinoma cell strain SGC 7901 was treated with MG 132 to inhibit its UPP specially. The effect of growth suppression on cells was evaluated with MTT assay. Cell cycle and apoptosis were detected by flow cytometry(FCM). DNA fragment analysis was used for confirming the presence of apoptosis. The activity of telomerase was examined by TRAP PCR ELISA. Expression of p27kip1 was detected by immunocytochemical technique. Results MG 132 had great inhibitory effect on the growth of SGC 7901 cells. The FCM analysis showed that the ratio of G0/G1 phase of control group was (46.3?4.1)%, the ratio of G0/G1 phase of SGC 7901 cells treated with MG 132 increased to (72.1?5.0)% ( P

Objective To explore the mechanisms of apoptosis induced by arsenic trioxide and amifostine in human acute promyelocytic leukemia cell lines HL-60 in vitro.Methods HL-60 cells were treated with different concentrations of arsenic trioxide alone and combined with amifostine.The inhibitory ratio of the ceils were measured by MTT assay.and the expression of Survivin Was detected by semiquantitate RT-PCR.Results Proliferation of HL-60 cells exposed to arsenic trioxide dwpped down with increasing dose of the dmg and this effect Was significantly hisher when arsenic trioxide Was used in combination with amifostine.Furthermore.there was a more significant decrease in Survivin expression in HL-60 cells treated with arsenic trioxide in combination with amifostine as compared to the cells treated only with arsenic trioxide.Conclusion Arsenic trioxide induced HL-60 cells to undergo apoptosis by downregulating the expression of Survivin. Amifostine enhanced the sensitivity of HL-60 cells to arsenic trioxide by downregulating the expression of Survivin,thus promoting apoptosis effect.

Objective To detect the reactive samples of enzyme-linked immunosorbent assay (ELISA1) by chemiluminescence microparticle immunoassay (CMIA),and to analyze the application value of CMIA in HCV infection validation of blood donors.Methods Nucleic acid 3-item combined testing (NAT),another ELISA2,HCV antibody supplementary test(Western Blot test,WB) and CMIA test supplemented in blood samples of 102 ELISA1 anti-HCV reactive blood donors were retrospectively analysed.Results Among 102 blood donors of anti-HCV positive,32 cases (31.37%,32/102) were HCV RNA reactive samples,50 cases (49.02%,50/102) were ELISA2/WB reactive simultaneously.With CMIA NAT results as the reference standard,CMIA was poorly correlated with HCV RNA (Spearman correlation coefficient rs=0.395,P<0.01),and the consistency between them was weak by Kappa test (Kappa=0.270,P<0.01).With ELISA2/WB detection results as the reference standard,CMIA was highly correlated with the results(Spearman correlation coefficient rs=0.713,P<0.01),and which showed high consistency by Kappa test (Kappa=0.674,P<0.01).Conclusion CMIA as a detection method of protein label after HCV infection has great value in the HCV infection confirmation in low-risk population.

Objective: To investigate the expression of peroxisome proliferator actived receptor ? (PPAR ?)and the inducement of apoptosis by PPAR ? ligand in renal cell carcinoma(RCC) derived cell lines.Methods:RT-PCR and Western blot analysis were performed to determined the expression of PPAR ? mRNA and protein in two RCC derived cell lines(786 O and A498) and two normal kidney(NK) derived cell lines(HK 2 and HMCC). Two RCC cell lines were treated with 50 ?mol/L troglitazoned for and evaluated for the effects of antidiabetic thiazolidinediones (TZDs) on the cells apoptosis by fluorescence microscopy and DNA ladder assay.The mutative expressions of Bcl 2 and Bax before and after TZDs treatment were also performed by western blot analysis. Results: The expression of PPAR ? was observed to be stronger in 786 O and A498 cells than in HK 2 and HMCC cells by RT-PCR and Western blot analysis. Treated with 50 ?mol/L troglitazone (for 48 h) it induced typical apoatosis in 786 O and A498 cells. After treatment, a decrease in Bcl 2 expression in RCC cells was observed by Western blot analysis,and the expression of Bax,however,was up regulated.Conclusion: The results reveal that troglitazone has the tumor suppressive effect on RCC cells. High affinity PPAR ? ligands (TZDs) may be the candidates for a novel approach to the treatment of this refractory neoplasm.

Objective:To express and purify a new fusion protein(RGD/tTF) for targeting therapy of cancer, and analyze its activities.Methods:The fused gene RGD/tTF was constructed by PCR, and then was inserted into vector pET22 b(+),expressed in E.coli BL21(DE3).The fusion protein was purified through Nickel-affinity chromatography column. The tTF activity of the fusion protein was analyzed by clotting assay and FⅩ activation assay. The specific binding of RGD/tTF to ?_v?_3 was analyzed by indirect ELISA.Results:The recombinant plasmid pET22 b(+)/RGD/tTF with correct sequence was obtained. The fusion protein was expressed with high yield in E.coli BL21(DE3). The purified fusion protein could activiate FⅩ and cause blood coagulation, and bind to ?_v?_3 specifically.Conclusion:The recombinant plasmid pET22 b(+)/RGD/tTF was constructed.The fusion protein retained TF activity and binding specificity to ?_v?_3, lays the foundation for studying the function of inducing thrombosis in tumor vasculature in vivo.