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Objective:To evaluate the diagnostic value of metagenomic next-generation sequencing (mNGS) in tuberculous meningitis (TBM).Methods:From August 1, 2020 to August 31, 2022, 99 patients with suspected TBM admitted to the Tuberculosis Diagnosis and Treatment Center, Hangzhou Chest Hospital, Zhejiang University School of Medicine were enrolled. The cerebrospinal fluid (CSF) was tested for mNGS, GeneXpert Mycobacterium tuberculosis/rifampin (GeneXpert MTB/RIF) and mycobacterium culture. The sensitivity, specificity, positive predictive value, negative predictive value, agreement rate, Kappa value of the diagnostic efficacy of the three test methods were compared.The receiver-operating characteristic (ROC) curve of the diagnostic efficacy of mNGS was drawn. Chi-square test and rank sum test were used for statistical analysis. Results:Among the 99 suspected patients with TBM, 67 were diagnosed with TBM and 32 were non-TBM. Based on the results of 67 cases clinically diagnosed with TBM, the sensitivity, specificity, positive predictive value, negative predictive value, agreement rate and Kappa value of mNGS for the diagnosis of TBM were 82.1%, 100.0%, 100.0%, 72.7%, 87.9% and 0.748, respectively. The sensitivity of mNGS was higher than that of GeneXpert MTB/RIF (50.7%) and mycobacterium culture (28.4%). The differences were statistically significant ( χ2=12.61 and 32.42, respectively, both P<0.01). The detection time of mNGS was 1.0 (1.0, 2.0) day, which was shorter than 42.0 (42.0, 42.0) days of mycobacterium culture with statistical significance ( Z=10.80, P<0.001). ROC curve analysis showed that mNGS had the best diagnostic efficacy when the number of Mycobacterium tuberculosis sequences in CSF was one. Conclusions:The sensitivity and specificity of mNGS in the diagnosis of TBM are high, and the detection time is shorter, which could be used in the early diagnosis of TBM.
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Objective:To investigate the changes of T-lymphocyte subsets, T-cell immunoglobulin and mucin domain molecule-1 (TIM-1) and TIM-3 levels, and cytokines in the peripheral blood of patients with active tuberculosis.Methods:From December 2017 to December 2018, 50 tuberculosis patients and 50 cured tuberculosis patients in Zhejiang Hospital of Integrated Chinese and Western Medicine were selected as the tuberculosis group and cured tuberculosis group, respectively. Fifty healthy individuals in the same period were selected as the control group. Flow cytometry was used to detect the T-lymphocyte subsets in the peripheral blood. The mRNA levels of TIM-1, TIM-3, interferon(IFN)-γ and interleukin(IL)-4 in peripheral blood mononuclear cells (PBMC) were detected by quantitative real-time polymerase chain reacticn (PCR). T test was used for statistical analysis. Results:The ratio of CD4 + /CD8 + T lymphocytes in the tuberculosis group (1.21±0.50) decreased significantly, comparing with those in the cured tuberculosis group (1.88±0.62) and the control group (1.92±0.82). The differences were statistically significant ( t=2.148 and 2.207, respectively, both P<0.05). The mRNA levels of TIM-1, TIM-3 and IL-4 in PBMC in the tuberculosis group were 2.16±0.37, 1.59±0.36 and 1.52±0.69, respectively, which were all higher than those in the cured tuberculosis group (1.60±1.23, 1.01±0.52 and 0.91±0.36, respectively) and the healthy control group (1.40±0.27, 0.92±0.34 and 0.79±0.42, respectively). All of these differences were statistically significant ( t=14.120, 11.440, 17.130, 12.090, 12.050 and 17.030, respectively, all P<0.05). However, the IFN-γ mRNA level (0.43±0.11) was lower than that in the cured tuberculosis group (1.74±0.72) and the control group (1.82±1.17), and the differences were both statistically significant ( t=13.880 and 11.430, respectively, both P<0.05). Conclusion:The immune dysfunction in patients with active tuberculosis may be related to the low ratio of CD4 + /CD8 + T lymphocytes, the increased expressions of TIM-1 and TIM-3, and the imbalance of helper T lymphocyte (Th)1/Th2 cytokines.
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Objective To investigate the effect of short course chemotherapy combined with broncho-vaxom on the prognosis of new smear positive pulmonary tuberculosis patients.Methods 80 patients with new smear positive pulmonary tuberculosis were selected as the research subjects.According to the random number table method,they were divided into observation group and control group,40 cases in each group.The control group was treated with 2HRZE/4HR chemotherapy,the observation group was treated with broncho-vaxom tablets based on the treatment of the control group.After 6 months of treatment,the sputum negative conversion rate,pulmonary lesions absorption,the incidence rate of adverse reaction and immune function were compared between the two groups.Results After 2,4,6 months of treatmentt,the sputum negative conversion rates of the observation group were 90.00％,95.00％ and 97.50％,respectively,which were significantly higher than those of the control group(x2 =4.020,4.114,3.914,all P ＜ 0.05).After 2,6 months of treatment,the total effective rates of the observation group were 87.50％ and 97.50％,which were significantly higher than those of the control group (x2 =4.588,5.000,all P ＜ 0.05).After 6 months of treatment,the IgA,IgG,IgM,CD3+,CD4+,CD4+/CD8+ in the observation group were (70.24 ± 6.19) ％,(46.89 ± 6.25) ％,(2.21 ± 0.39),(3.86 ± 1.43) g/L,(14.76 ± 2.58) g/L,(1.47 ± 0.65) g/L,respectively,which were significantly higher than those of the control group(t =-2.116,-2.575,-2.322,-2.138,-4.513,-2.599,all P ＜ 0.05),the CD8+ was (18.85 ± 2.08) ％,which was significantly lower than that in the control group (t =2.609,P ＜ 0.05).There was no statistically significant difference in the incidence of adverse reactions between the two groups (x2 =0.251,P ＞ 0.05).Conclusion Short course chemotherapy regimen combined with bronchovaxom can improve the new smear positive pulmonary tuberculosis patients with recent sputum negative conversion rate and lung lesions absorption effect,improve the immune function of patients and without increasing the incidence of adverse reactions,it is worthy of clinical attention.