Objective To prepare a novel monoclonal antibody (mAb) that specifically against Mycobacterium tuberculosis culture filtrate protein 10 (CFP-10).Methods The BALB/c mice were immunized by a peptide with 14 amino acids (aa residues 53 to 66) of CFP-10,and then the splenocytes of mice were fused with myeloma cell line SP2/0.The resultant fused cells were subjected to screening culture,enzyme linked immunosorbent assay (ELISA) assay and subcloning by limited dilution to establish hybridoma cell lines of stable secreting anti-the peptide of CFP-10 antibody.The antibody was purified,and its isotypes were analyzed.Then,the antibody was further evaluated by Western blotting,immunoprecipitation and ELISA in 38 culture supernatant samples of Mycobacterium tuberculosis,20 culture supernatant samples of non-Mycobacterium tuberculosis,32 samples of tuberculous pleural effusion,24 samples of non-tuberculous pleural effusion,and 20 serum samples from healthy controls.Results The isotype of the mAb against the specific peptide of CFP-10 was an IgG1 with κ chain,and it was applicable for Western blotting and immunoprecipitation analysis.ELISA quantitative test showed that the sensitivity and specificity for diagnosis of Mycobacterium tuberculosis were 78.6％ (55/70) and 92.2％ (59/64),respectively.Conclusion The mAb generated against the specific peptide of CFP-10 is high in sensitivity and specificity,and it might be used in the early diagnosis of tuberculosis.

Objective A recombinant adenoviral vector containing mIL-18BP and mIL-4 fusion gene(AdmIL-18BP/mIL-4) was constructed and used to investigate the role of mIL-18BP and mIL-4 in medula-ring the expressions of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthetase (iNOS) and their inducing products PGE2, NO in murine collagen-induced arthritis. Methods Male DBA-1/BOM mice were used in this study. Mice with CIA were intra-articularly injected with 107 pfu/6μl of AdmIL-18BP/mIL-4.Intra-articular injections of AdLacZ or PBS were used as controls. The mRNA expression of COX-2, iNOS in synovial tissue was analyzed by semi-quantitative RT-PCR. Expression of COX-2 and iNOS protein was estimated by Western blot method. The production of PGE2 and NO in synovia was detected by competitive ELISA and enzyme reduction of nitrate. Results The expression of COX-2, iNOS mRNA in routine synovial tissue of AdmIL-18BP/mIL-4 treatment group was significantly lower than that of AdLacZ group (0.15 vs 0.42,P<0.01 ; 0.05 vs 0.77, P<0.01) and PBS group (0.15 vs 0.65, P<0.01; 0.05 vs 0.64, P<0.01 ). And the protein expression of COX-2, iNOS from AdmIL-18BP/mIL-4 treatment group was also obviously lower than that of AdLacZ group (0.08 vs 0.92, P<0.01; 0.11 vs 1.00, P<0.01) and PBS group (0.08 vs 0.77, P<0.01; 0.11 vs 0.84, P<0.01 ). The PGE2 and NO production in synovia of AdmIL-18BP/mIL-4 treatment group was significantly lower than that of AdLacZ group [(0.68x0.06) vs (2.58±0.21)ng/mL, P<0.01; (23.4+2.5) vs (60.0±11.3)μmol/L, P<0.01 ] and PBS group [(0.68±0.06) vs (2.57±0.20)ng/mL, P<0.01; (23.4+2.5) vs (60.3±13.4)μmol/L, P<0.01]. Conclusion These data indicat that local over-expre-ssion of mIL-18BP and mIL-4 can down-regulate COX-2, iNOS and their induced product PGE2, NO in CIA mice. The combination treatment with mIL-18BP and mIL-4 is a promising therapeutic target for RA.

Objective:To investigate the clinical application value of serum carcinoembryonic antigen (CEA) level in predicting lymph node metastasis of resectable medullary thyroid carcinoma (MTC) .Methods:140 patients of resectable MTC from Zhejiang Cancer Hospital and Hangzhou First People’s Hospital from Jan. 2009 to Feb. 2019 were included. The relationship of serum CEA and lymphatic metastasis was retrospectively analyzed in 140 patients of resectable MTC, and the clinical significance of serum CEA for predicting total lymph node, central lymph node, lateral lymph node and upper mediastinal lymph node metastasis was also evaluated.Results:The positive rate of serum CEA in resectable MTC was 77.14%. The expression level of serum CEA in resectable MTC with lymph node metastasis was significantly higher than those without lymph node metastasis ( P<0.001) . Spearman correlation analysis further indicated that the level of serum CEA expression was positively correlated with the number of lymph node metastases of resectable MTC patients ( P<0.001) . The area under curve of predicting lymphatic metastasis of total lymph node, central lymph node, lateral lymph node and upper mediastinum was 0.773, 0.768, 0.827 and 0.847. When the cut-off value of serum CEA was 6.58, 11.43, 15.74 and 30.45 ng/ml, respectively, the sensitivity of serum CEA to predict total, central, lateral neck and upper mediastinal lymph node metastasis was 88.46%, 81.43%, 85.00%, 95.00%, and the specificity was 56.45%, 60.00%, 71.25%, and 69.17%, respectively. Conclusion:Serum CEA has a high positive expression rate in resectable MTC, and its expression level has important clinical significance in evaluation of lymphatic metastasis.