Objecave To develop a sensitive and specific LC-MS/MS method for determination of testolactone in human urine.Methods A C_(18 )column(2.1×50mm,3.5μm) was used.The mobile phase Was a mixture of acetonitrile and the buffer solution(ammonium acetate-water solution adjusted with formic acid to pH 3.5)at a flow rate of 0.5ml/min.A mass spectrometer equipped with electrospray ionization source was used as a detector and operated in the positive mode.In multiple reaction monitoring(MRM)mode,the ion transitions of m/z 301→121 and m/z 301→25 was used to qualify and quantify the testolactone,respectively.Results Chromatograms showed no endogenous interfering peaks with the urine blank sample.Each analysis was completed within 7min The calibration wag linear in the concentration range within 0.1～50μg/ml.The intra-batch and inter-batch RSD were less than 10%.The recovery rate of the extraction was about 60%.Conclusions The method is proved to meet the requirements of WADA and be suitable for routine screening.

Objective To investigate the relationship between cellular immune response which is related with Th1/Th2 cells and the different severities of tuberculosis pleural effusion adhesion.Methods A total of 66 in-patients diagnosed with different severities of tuberculosis pleural effusion adhesion by internal thoracoscope were enrolled from August 2014 to December 2016.ELISA was used to determine levels of INF-γ,TNF-α,IL-2 and IL-4. The ratio of Th1 and Th2 cells was detected by flow cytometry. Results All the patients were divided into 3 groups:9 cases without pleural adhesion,32 cases with mild pleural adhesion,25 cases with severe pleural adhe-sion. The levels of 4 cytokines in pleural fluid were significantly higher than those in serum in each group(P <0.05,respectively).The concentrations of INF-γ and TNF-α were increased with the severity of tuberculosis pleu-ral effusion adhesion. The levels of INF-γ and TNF-α in the severe pleural adhesion group were markedly higher than those in the other two groups(P<0.05,respectively).The proportion of Th1 cells in the severe pleural adhe-sion group was significantly higher than that in the none pleural adhesion group and in the mild pleural adhesion group(P<0.05,respectively).Conclusions The proportion of Th1 cells and levels of INF-γ and TNF-α are pos-itively related with pleural adhesion severity. Cellular immune response which is related with Th1 cells contributes to pathological immune pleural damage and intensify the severity of pleural adhesions.

Objective To compare the concentrations of alpha-enolase (ENO1),CYFRA21-1,and CA125 in the patients with malignant pleural effusion ,tuberculous exudative pleural effusion ,or parapneumonia pleural effusion. To explore the clinical value of ENO1 in pleural effusion combined with serum CYFRA21-1 and CA125 in diagnosis of malignant pleural effusion. Methods Enzyme-linked immunosorbent assay(ELISA)was used to detect the concentration of ENO1 in pleural effusions. The concentrations of CA125 and CYFRA21-1 in the blood samples were measured using chemiluminescence and magnetic particle-based chemiluminescence respective-ly. The sensitivity and specificity of ENO1 combined with serum CYFRA21-1 and CA125 detection were calculated. Results The concentration of ENO1 in malignant pleural effusion group was significantly increased(P<0.001);the concentrations of ENO1 did not differ significantly between tuberculous exudative pleural effusion and parapneu-monia pleural effusion(P>0.05). The sensitivity and specificity of ENO1 combined with serum CYFRA21-1 and CA125 detection in malignant pleural effusion were 94％ and 74％,98％ and 98％,respectively. Conclusions ENO1 combined with serum CYFRA21-1 and CA125 detection can improve the sensitivity of diagnosis of malignant pleural effusion and enhance the diagnostic rate of malignant pleural effusion.