The biotin synthesis pathway in Mycobacteria tuberculosis is a new target for the development of anti-tuberculosis drugs / 药学学报

italic>Mycobacterium tuberculosis, responsible for tuberculosis (TB), remains a major health problem worldwide and is one of the infectious diseases causing increased morbidity and mortality worldwide. Biotin, namely vitamin H, is an important cofactor necessary for fatty acid biosynthesis, gluconeogenesis and amino acid metabolism in organisms including Mycobacterium tuberculosis. Due to its inability to ingestion biotin from outside, Mycobacterium tuberculosis can only obtain biotin through biotin biosynthesis. Different from the classical BioC-BioH, BioI-BioW and non-classical BioZ pathways, Mycobacterium tuberculosis synthesized biotin by "BioC-BioH(2)" pathway in the early stage. This review focuses on the unique biotin synthesis pathway of Mycobacterium tuberculosis and its key genes, especially the response of this pathway and biotin-dependent carboxylase to tuberculosis first-and second-line drugs, as well as inhibitors and natural products targeting biotin synthesis.

Drug resistance surveillance in Mycobacterium tuberculosis in Jiaxing City / 预防医学

Objective@#To investigate the resistance of Mycobacterium tuberculosis to first-line anti-tuberculosis drugs in Jiaxing City, Zhejiang Province from 2017 to 2019, so as to provide insights into improvements of the therapeutic effect of pulmonary tuberculosis. @*Methods@#Data pertaining to pulmonary tuberculosis in Jiaxing City from 2017 to 2019 were collected from the Tuberculosis Surveillance System of Chinese Disease Prevention and Control Information System, including demographics, treatment classification, sputum culture and drug resistance. The spectrum, types and prevalence of drug resistance in M. tuberculosis to four first-line tuberculosis drugs, including isoniazid (INH), rifampicin (RFP), streptomycin (SM) and ethambutol (EMB), was analyzed using a descriptive epidemiological method.@*Results@#A total of 1 310 M. tuberculosis isolates were cultured from pulmonary tuberculosis patients in Jiaxing City from 2017 to 2019, and there were 259 M. tuberculosis isolates that were resistant to anti-tuberculosis drugs, with an overall drug resistance rate of 19.77%. The prevalence rates of drug resistance to INH, SM, RFP and EMB were 13.36%, 11.83%, 5.50% and 3.59%, respectively. The prevalence of drug resistance was lower in M. tuberculosis isolates from treatment-naïve patients than from retreated patients (18.45% vs. 34.58%, P<0.05). M. tuberculosis isolates presented high resistance to SM (4.50%) and INH alone (4.35%), the highest resistance to INH-SM combinations (3.28%), and the highest resistance to INH+RFP+SM combinations (1.83%). Sixteen isolates were resistant to all the four drugs, with a drug resistance rate of 1.22%. The proportions of resistance to a single drug, RFP resistance, multidrug resistance and resistance to two and more drugs were 10.31%, 5.50%, 4.73% and 4.73%, respectively. In addition, the prevalence of RFP resistance among all patients and treatment-naïve patients both showed a tendency towards a rise from 2017 to 2019 (P<0.05). The prevalence of RFP resistance (7.01% vs. 3.76%) and resistance to two and more drugs (6.01% vs. 3.25%) was both higher among interprovincial mobile tuberculosis patients than among local non-mobile patients (P<0.05). @*Conclusions@#The overall prevalence of drug resistance was lower in M. tuberculosis isolates in Jiaxing City from 2017 to 2019 than in Zhejiang Province, with INH and RFP resistance as predominant types.

Transcriptional analysis on biomarkers of liver injury induced by anti-tuberculosis drugs / 上海预防医学

ObjectiveThe study utilized human transcriptome microarray to explore biomarkers for diagnosing drug-induced liver injury （DILI） caused by anti-tuberculosis drugs. MethodsA 6-month follow-up study was conducted on 152 patients treated with anti-tuberculosis drugs in designated hospitals in Shanghai. The blood samples were collected at the 0， 2， 4， 8， 12 and 24 weeks after treatment. According to the clinical biochemical indicators， the research subjects were divided into DILI cases （34 cases） and Control cases （118 cases）. Single factor analysis was conducted on the influencing factors between the two groups. In a 1∶1 matched DILI-control study， RNA samples of 13 pairs of cases were sequenced by the whole transcript expression mRNA array. Differentially expressed genes （DEGs） were screened by Hotelling's T2 value sequencing and the expression trend analysis of genes by STEM （short-time series expression miner）， and the functional enrichment and pathway analysis of DEGs were carried out. ResultsIn total 152 clinical cases， weight of patients was a risk factor for the occurrence of hepatotoxicity caused by anti-tuberculous drugs. Based on the analysis results of mRNA array， 513 DEGs were screened by Hotelling's T2 value sequencing method， which were enriched in 32 annotations of GO （Gene Ontology） analysis and 10 pathways of KEGG （Kyoto encyclopedia of genes and genomes） analysis. One differential expression pattern was screened by STEM， which was enriched in 2 biological process notes of GO. Among them， the key genes AIM2， CD86， CXCL10 and non-coding RNAs SCARNA10， SNHG10 and SNORD105 are potential biomarkers of DILI caused by anti-tuberculosis drugs. ConclusionIn this research for biomarkers conducted on cases with liver injury caused by anti-tuberculosis drugs， biological pathways associated with hepatotoxicity are identified and a series of key genes related with drug-induced liver injury are found， which provides the basis for mechanism study and searching for earlier and more sensitive biomarkers.

Study on the Prevention Mechanism of Anti-tuberculosis Drug-induced Liver Injury with Orazamide Based on HMGB1-RAGE Signaling Pathway / 中国药房

OBJECTIVE：To prelimi narily investigate the possible mechanism of orazamide to prevent anti-tuberculosis drug-induced liver injury （ATB-DILI）. METHODS ：A total of 60 Kunming mice were randomly divided into blank group ，model group，positive control group [diammonium glycyrrhizinate 60 mg/（kg·d）]，orazamide low-dose ，medium-dose and high-dose groups [ 80，160，320 mg/（kg·d）]，with 10 mice in each group. Except for blank group ，other groups were given isoniazid [ 75 mg/（kg·d）]+rifampicin [ 75 mg/（kg·d）] for 14 days intragastrically to induce ATB-DILI model. At the same time ，administration groups were given relevant medicine intragastrically ，blank group and model group were given normal saline intragastrically. The administration volume was 20 mL/（kg·d），once a day ，for consecutive 14 days. The general conditions of the mice were observed and recorded every day ，such as growth and development ，mental and diet state. After last medication ，liver index was calculated ， and HE staining was adopted to observe pathological changes of liver tissue of mice. The positive expression of high mobility group protein B 1 （HMGB1） and NF-κ B in liver tissue were detected by streptavidin biotin-peroxidase complex （SABC） immuno- histochemistry. The serum levels of liver function indexes in serum ，the protein expression of advanced glycation end product receptor（RAGE）and TNF-α in liver tissue were detected by ELISA. RESULTS：Compared with blank group ，the growth and development of mice in the model group were slow ，and their appetite and spirit were poor. The liver index ，serum levels of TBIL ， DBIL，ALT，AST，ALP，TBA and γ-GT were increased significantly （P＜0.05）. Structural disorder of liver lobules ，degeneration and necrosis of liver cells and inflammatory cell infiltration were observed. The expression of HMGB 1，NF-κB，RAGE and TNF-α in liver tissue were elevated significantly （P＜0.05）. Compared with model group ，the general condition of mice were all improved to different extents in orazamide low-dose ，medium-dose and high-dose groups ，positive control group ，while liver index and above serum indexes were all decreased significantly （P＜0.05）. The pathological changes of liver tissue were all improved to different extents ，while the protein expression of HMGB 1，NF-κB，RAGE and TNF-α were all decreased significantly（P＜0.05）. The improvement of above indexes in orazamide high-dose group were all significantly better than orazamide low-dose and medium-dose groups （P＜0.05）；the levels of ALP and TBA in orazamide high-dose group were significantly lower than positive control group （P＜0.05）. CONCLUSIONS ：Orazamide can prevent ATB-DILI induced by isoniazid combined with rifampicin in mice，the mechanism of which may be associated with down-regulating the protein expression of HMGB 1 and RAGE in liver tissue and inhibiting the secretion of inflammatory factors.

Study on CYP2E1 gene polymorphism and the relationship between methylation level of its promoter region and liver injury induced by anti-tuberculosis drugs in Mongolian tuberculosis patients in Inner Mongolia / 医学研究生学报

ObjectiveThere are few studies on whether the occurrence of anti-tuberculosis drug-induced liver injury (ADIH) is associated with the polymorphism of CYP2E gene and methylation level. This study aims to CYP2E1 gene polymorphism and the relationship between the methylation level of the promoter region and ADIH in Mongolian tuberculosis (TB) patients.Methods A total of 135 Mongolian TB patients who received standardized treatment at the Tuberculosis Research Institute of Tongliao City, Inner Mongolia from November 2015 to June 2018 were selected. According to the ADIH criteria, TB patients with liver injury were selected as the ADIH group (n=45), and TB patients without liver injury were matched as the control group based on a ratio of 1∶2 (n=90). DNA extraction and polymerase chain reaction (PCR) were performed to amplify the CYP2E1 gene to determine the CYP2E1 rs2031920 genotype, and to analyze the CYP2E1 gene polymorphism and relationship between ADIH and promoter methylation level.Results There were no significant differences in the distribution of CYP2E1 rs2031920 genotype, C1 and C2 gene frequencies between the ADIH group and the control group (P>0.05). The overall methylation level in the promoter region of CYP2E1 gene in ADIH group (0.711±0.085) was significantly lower than that of the control group (0.759±0.062). Results of Logistic regression showed that the overall methylation level in the promoter region of CYP2E1 gene was the influencing factor for the occurrence of ADIH (P<0.005). For each 0.1 unit increase of methylation level, the risk of ADIH occurrence reduced by 0.388 times, and the OR (95% CI) value was 0.388 (between 0.204 and 0.739).Conclusion The overall methylation level in the promoter region of CYP2E1 gene was reduced in Mongolian ADIH patients, but the polymorphism of CYP2E1 gene was not related to the occurrence of ADIH. These results suggested that CYP2E1 methylation could be applied to the prevention and treatment of ADIH in patients with tuberculosis.

Study on CYP2E1 gene polymorphism and the relationship between methylation level of its promoter region and liver injury induced by anti-tuberculosis drugs in Mongolian tuberculosis patients in Inner Mongolia / 医学研究生学报

ObjectiveThere are few studies on whether the occurrence of anti-tuberculosis drug-induced liver injury (ADIH) is associated with the polymorphism of CYP2E gene and methylation level. This study aims to CYP2E1 gene polymorphism and the relationship between the methylation level of the promoter region and ADIH in Mongolian tuberculosis (TB) patients.Methods A total of 135 Mongolian TB patients who received standardized treatment at the Tuberculosis Research Institute of Tongliao City, Inner Mongolia from November 2015 to June 2018 were selected. According to the ADIH criteria, TB patients with liver injury were selected as the ADIH group (n=45), and TB patients without liver injury were matched as the control group based on a ratio of 1∶2 (n=90). DNA extraction and polymerase chain reaction (PCR) were performed to amplify the CYP2E1 gene to determine the CYP2E1 rs2031920 genotype, and to analyze the CYP2E1 gene polymorphism and relationship between ADIH and promoter methylation level.Results There were no significant differences in the distribution of CYP2E1 rs2031920 genotype, C1 and C2 gene frequencies between the ADIH group and the control group (P>0.05). The overall methylation level in the promoter region of CYP2E1 gene in ADIH group (0.711±0.085) was significantly lower than that of the control group (0.759±0.062). Results of Logistic regression showed that the overall methylation level in the promoter region of CYP2E1 gene was the influencing factor for the occurrence of ADIH (P<0.005). For each 0.1 unit increase of methylation level, the risk of ADIH occurrence reduced by 0.388 times, and the OR (95% CI) value was 0.388 (between 0.204 and 0.739).Conclusion The overall methylation level in the promoter region of CYP2E1 gene was reduced in Mongolian ADIH patients, but the polymorphism of CYP2E1 gene was not related to the occurrence of ADIH. These results suggested that CYP2E1 methylation could be applied to the prevention and treatment of ADIH in patients with tuberculosis.

Effect of first-line antituberculous treatment on vitamin D level in patients with pulmonary tuberculosis / 中华临床营养杂志

Objective To explore the effect of first-line anti-tuberculosis treatment on vitamin D level in patients with pulmonary tuberculosis,and to master the changes of vitamin D level in the course of treatment,so as to provide a scientific basis for tuberculosis and nutrition health education in Shenzhen.Methods A total of 100 patients diagnosed as smear-positive pulmonary tuberculosis and receiving initial treatment in 2016 were enrolled and all the patients were treated with the standardized short-course chemotherapy regimens.The blood samples were extracted before treatment and at the ends of intensive and continuation phase.The 25-hydroxyvitamin D [25-(OH) D] concentrations were determined by chemiluminescence (CLIA) at each time point.The change of 25-(OH) D concentrations during anti-tuberculosis treatment was analyzed and the differences of vitamin D levels between different time points were identified.Results 79 (79.0％),94 (94.0％) and 96 (96.0％) patients were found vitamin D deficiency before treatment and at the end of the intensive and continuation phases respectively,which showed an upward trend (x2=15.543,P＜0.001) and the 25-(OH)D concentrations were (15.74±6.54) ng/ml,(12.56±5.15) ng/ml,(11.51±4.28) ng/ml,respectively.During the whole course of treatment,the 25-(OH) D concentration decreased by 26.9％ or (4.23 ± 6.75) ng/ml (t =6.257,P＜0.001),wherein it decreased (3.18 ± 5.24) ng/ml in intensive phase (t =6.069,P＜ 0.001) and (1.05±4.86) ng/ml in continuation phase (t =2.154,P =0.034).The former had a greater decreased value (t=2.836,P=0.006).There were 77 (77.0％) and 55 (55.0％) patients with 25-(OH)D concentration reduction in intensive and continuation phases respectively (x2 =9.680,P =0.003),of which 41 patients (41.0％) continued to decline.Conclusion Once anti-tuberculosis treatment is conducted,the vitamin D level will decrease rapidly in the intensive phase and continue decreasing throughout the course of treatment,which leads to a general lack of vitamin D in patients with primary pulmonary tuberculosis.First-line anti-tuberculosis drugs may be the main cause for vitamin D level reduction.Therefore,it is necessary for clinicians to strengthen vitamin D health education for each patient throughout the treatment period,especially for those at high risk of vitamin D deficiency who should be recommended adjuvant vitamin D supplementation therapy.

Application of Mycobacterium marinum infected zebrafish for evaluation of anti-tuberculosis drugs / 药学学报

Tuberculosis (TB) is a serious infectious disease caused by Mycobacterium. tuberculosis. In recent years, with the emergence of drug-resistant forms, the development of new anti-tuberculosis drugs is urgently needed. In this study, we used Mycobacterium marinum (M. marinum), which is highly similar to M. tuberculosis, to establish a M. marinum infected-zebrafish model and quantitative PCR (qPCR) method for bacterial count analysis. The results showed that injecting M. marinum into the yolk sac is an efficient and convenient way to infect zebrafish embryos. By counting the survival rate of infected zebrafish and the number of bacteria in zebrafish by Ziehl-Neelsen staining, we analyzed the efficacy of isoniazid and rifampicin as anti-tuberculosis drugs and the synergistic effect of drugs. The results suggested that three evaluation methods exhibit good consistency. This study demonstrated that zebrafish-M. marinum infection model combined with qPCR analysis is a simple and efficient method for in vivo screening and evaluation of anti-tuberculosis drugs. Animal experiments were carried out in accordance with the provisions for animal ethics in the Regulations on Laboratory Animals of Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences.

Anti-tuberculosis drug-induced liver injury correlated with the methylation in the promoter regions of CYP2E1 and GSTM1 / 医学研究生学报

Objective No study has been reported on the association between the abnormal methylation of drug metabolic enzymes and anti-tuberculosis drug-induced liver injury (ATLI). This article aimed to investigate the relationship of ATLI with the methylation of the CpG islands in the promoter regions of cytochrome P450 2E1 (CYP2E1) and glutathione s-transferase M1 (GSTM1) in Chinese Mongolian patients with tuberculosis (TB). Methods This retrospective study included 93 cases of TB diagnosed and treated in the TB prevention and treatment institutions of Tongliao, Inner Mongolia, between September 2016 and December 2017, which were divided into an ATLI (n = 31) and a non-ATLI group (n = 62), the former with and the latter without ATLI within 6 months after anti-TB medication. We compared the methylation levels of the CYP2E1 and GSTM1 genes between the two groups of patients and analyzed the risk factors of ATLI. Results In comparison with the non-ATLI controls, the patients of the ATLI group showed significantly lower methylation levels in the promoter regions of CYP2E1 (0.759 ± 0.066 vs 0.694 ± 0.091, P < 0.05) and GSTM1 (0.207 ± 0.093 vs 0.187 ± 0.092, P < 0.05). Multivariate logistic regression analysis revealed that the main risk factors of ATLI included alcohol consumption (OR = 5.329, 95% CI: 1.442-19.697, P < 0.05) and methylation in the CYP2E1 promoter region (OR = 0.312, 95% CI: 0.165-0.591, P < 0.05) in the TB patients. Conclusion ATLI is associated with the methylation level in the promoter region of the CYP2E1 gene in Chinese Mongolian patients with tuberculosis, indicating that the methylation of CYP2E1 could be used as a biomarker in the prevention and control of ATLI.

Identification of novel anti-tuberculosis lead compound targeting shikimate kinase / 药学学报

Shikimate kinase is a key protein of the shikimic pathway, which is essential for the survival of Mycobacterium tuberculosis. In this study, a screening assay for Mycobacterium tuberculosis shikimate ki-nase (MtSK) inhibitor was developed. A 120 000-compound library was screened by the enzyme assay and the phenotype screening using Mycobacterium smegmatis. A hit compound named IMB-T5297[(E)-3-(3-(3- chloro-5-methoxy-4-(prop-2-yn-1-yloxy)phenyl)acryloyl)-6-methyl-2H-pyran-2,4(3H)-dione] was identified to be a selective inhibitor of MtSK with a half maximal inhibitory concentration (IC50) value of 1.745 μg·mL-1, which also showed antibacterial activity. The interaction between compound and protein was analyzed by surface plasmon resonance (SPR) experiment, which showed the KD value was 2.151×10-5 mol·L-1. The binding model of MtSK and compound was simulated by the computer program. Five key amino acids in the binding pocket were indispensable site-directed mutated to verify the model. IMB-T5297 inhibited Mycobacterium tuberculosis H37Rv with a minimum inhibitory concentration (MIC) value of 49.723 μg·mL-1 and displayed low cytotoxicity to mammalian cells. In this study, IMB-T5297 was identified as a selective inhibitor of MtSK enzyme with anti-tuberculosis activity. With additional structural modification, the compound has a potential to become a novel anti-tuberculosis compound.

Expression levels of serum nitric oxide and nitric oxide synthase intheUy-ghur and Han nationality patients with anti-tuberculosis drug-induced liver injury in Xinjiang / 中国感染控制杂志

Objective To explore the expression levels of serum nitric oxide(NO)and nitric oxide synthase(NOS) between Han and Uyghur nationality patients with anti-tuberculosis drug-induced liver injury(ATDLI). Methods Patients with confirmed ATDLI in Chest Hospital of Xinjiang Uyghur Autonomous Region and First Affiliated Hos-pital of the Medical College of Shihezi University between January 2015 and May 2016 were chosen and divided into Han group and Uyghur group.By detecting the expression levels of NO and NOS in serum of ATDLI patients,ex-pression levels of serum NO and NOS in ATDLI patients of different gender,body mass index(BMI),and liver function injury were compared.Results 100 ATDLI patients in Han group and 135 in Uyghur group were recruited in study. Expression levels of NO and TNOS in Han group were(134.24±27.60)μmol/L and(33.01 ±4.23)U/mL respectively,in Uyghur groups were(97.10±17.41)μmol/L and(27.41 ±3.95)U/mL respectively,serum levels of NO,TNOS,iNOS,and eNOS in Han patients were all higher than Uyghur patients,difference was statistically significant(P<0.01). In Han ATDLI group,serum levels of NO and TNOS in male patients were both higher than female patients(P<0.05);in Uyghur ATDLI group,serum levels of NO,TNOS,and iNOS in male patients were all higher than female patients(P<0.01). The expression levels of serum NO,TNOS,and iNOS of Han group were all higher than the same gender in Uyghur group(P<0.001),difference in levels of NO and TNOS among different body mass index(BMI)groups in Han and Uyghur patients were both statistically significant(P<0.01). In both group,levels of NO and TNOS in obese patients were both higher than lean patients and normal weight patients(P<0.05). The correlation analysis showed that NO levels of Han and Uyghur groups were both positively correlated with BMI(r=0.444,0.677,respectively,P<0.01). There were significant differences in serum NO and NOS levels between Han and Uyghur patients with different degrees of liver injury(P<0.05);NO and NOS levels in both groups with mild liver injury were both lower than those with moderate and severe injury (P<0.001).Conclusion Serum NO and NOS levels between ATDLI Han group and Uyghur group are different,serum NO level is related to BMI,and it can increase with the degree of liver injury.

Pyrrolidine dithiocarbamate alleviates the anti-tuberculosis drug-induced liver injury through JAK2/STAT3 signaling pathway: An experimental study

OBJECTIVE@#To study the effect of pyrrolidine dithiocarbamate (PDTC) on the anti-tuberculosis drug-induced liver injury and the molecular mechanism.@*METHODS@#Clean male SD rats were selected as experimental animals and randomly divided into normal group, model group, PDTC group and AG490 group. Animal model of anti-tuberculosis drug-induced liver injury was established by intragastric administration isoniazid + rifampicin. PDTC group received intraperitoneal injection of PDTC, and AG490 group received intraperitoneal injection of AG490. Twenty-eight days after intervention, the rats were executed, and the liver injury indexes, inflammation indexes and oxidative stress indexes in serum as well as JAK2/STAT3 expression, liver injury indexes, inflammation indexes and oxidative stress indexes in liver tissue were determined.@*RESULTS@#p-JAK2, p-STAT3, TNF-α, IL-1β, IL-6, ROS, 8-OHdG and MDA expression in liver tissue as well as TBIL, ALT, AST, γ-GT, TNF-α, IL-1β, IL-6, 8-OHdG and MDA levels in serum of model group were significantly higher than those of normal group while p-JAK2, p-STAT3, TNF-α, IL-1β, IL-6, ROS, 8-OHdG and MDA expression in liver tissue as well as TBIL, ALT, AST, γ-GT, TNF-α, IL-1β, IL-6, 8-OHdG and MDA levels in serum of PDTC group and AG490 group were significantly lower than those of model group.@*CONCLUSIONS@#PDTC can inhibit the inflammation and oxidative stress mediated by JAK2/STAT3 signaling pathway to alleviate the anti-tuberculosis drug-induced liver injury.

Expression of microRNA-3620 in blood plasma and its clinical significance in patients with anti-tuberculosis drug-induced hepatotoxicity / 中华传染病杂志

Objective To investigate the expression of microRNA (miRNA)-3620 in the plasma of patients with anti-tuberculosis drug-induced hepatotoxicity (ATDH).Methods A total of 35 patients with ATDH and 35 non-ATDH paired individuals were included in this study.Plasma miRNA-3620 levels were detected using real-time Polymerase chain reaction.Comparison between two groups was done with t test.Receiver operation characteristic (ROC) curve analysis was performed to determine the diagnostic value of miRNA-3620 in ATDH.Results The relative expression of plasma miRNA-3620 of patients with ATDH and non-ATDH were 1.65±1.43 and 0.71±0.45, respectively, with significantly statistical difference (t=3.703, P<0.01).The cut off value of miRNA-3620 expression was 1.15 and the area under ROC curve were 0.71(95% CI: 0.43-1.45).Based on this cutoff value, the sensitivity and specificity of miRNA-3620 in diagnosing ATDH were 60.0% and 82.9%, respectively;the positive predictive value was 77.8% and the negative predictive value was 67.4%.Twenty-one ATDH cases and 29 non-ATDH cases was correctly diagnosed, with the accuracy of 71.4%.Conclusion The expression of miRNA-3620 in plasma is significantly increased in ATDH patients.

Characteristics of drug-resistant Mycobacterium tuberculosis and new anti-tuberculosis drugs:research advances / 国际药学研究杂志

The growing epidemic of drug-resistant tuberculosis has posed a great threat to people′s health and become a major public health and social problems. Early detection and standardized treatment of tuberculosis are of great significance to controlling its spread. In order to provide reference for better prevention and clinical treatment of tuberculosis,this paper reviews the current situa-tion of anti-tuberculosis drugs and mechanism of resistance and new potential drugs for treating tuberculosis.

A retrospective study on primary anti-tuberculosis drug resistance in patients with acquired immunodeficiency syndrome and tuberculosis / 中华传染病杂志

Objective To investigate primary anti-tuberculosis drug resistance in patients with acquired immunodeficiency syndrome (AIDS) and tuberculosis in Chongqing area.Methods Clinical data of 119 patients with AIDS and tuberculosis were retrospectively collected.Anti-tuberculosis drug resistance rates were analyzed according to drug susceptibility testing, and their correlations with CD4+ T lymphocytes counts, initially treatment or retreatment and clinical forms of tuberculosis were also analyzed.Comparison between groups was analyzed by x2 test.Results Thirty-eight patients (31.9%) showed anti-tuberculosis drug resistance among the 119 patients with completed results of drug susceptibility testing results.The percentages of mono-resistance, poly-resistance, multi-drug resistance (MDR) and extensive drug resistance (XDR) were 11.7%, 7.6%, 6.7% and 5.9%, respectively.The resistance rate of isoniazid (22.7%, 28/119) was the highest among first-line anti-tuberculosis drugs and that of pasiniazide (11.0%, 14/119) was the highest among second-line drugs.Drug resistance rates among patients with different levels of CD4+ T lymphocytes counts did not differ significantly (the cut-off of CD4+ T lymphocytes count was 50/μL: x2=0.545, P=0.461;cut-off value was 100/μL: x2=0.652, P=0.420).Patents with milliary pulmonary tuberculosis had a significantly higher drug resistance rate (64.0%) than those with secondary pulmonary tuberculosis (27.6%).Conclusions The prevalence of anti-tuberculosis drug resistance prior to anti-tuberculosis treatment initiation is high among AIDS patients with tuberculosis in Chongqing area.Patients with milliary pulmonary tuberculosis tend to have higher anti-tuberculosis drug resistance, but drug resistance does not appear to correlate with CD4+ T lymphocytes counts.

Pyrrolidine dithiocarbamate alleviates the anti-tuberculosis drug-induced liver injury through JAK2/STAT3 signaling pathway: An experimental study

Objective To study the effect of pyrrolidine dithiocarbamate (PDTC) on the anti-tuberculosis drug-induced liver injury and the molecular mechanism. Methods Clean male SD rats were selected as experimental animals and randomly divided into normal group, model group, PDTC group and AG490 group. Animal model of anti-tuberculosis drug-induced liver injury was established by intragastric administration isoniazid + rifampicin. PDTC group received intraperitoneal injection of PDTC, and AG490 group received intraperitoneal injection of AG490. Twenty-eight days after intervention, the rats were executed, and the liver injury indexes, inflammation indexes and oxidative stress indexes in serum as well as JAK2/STAT3 expression, liver injury indexes, inflammation indexes and oxidative stress indexes in liver tissue were determined. Results p-JAK2, p-STAT3, TNF-α, IL-1β, IL-6, ROS, 8-OHdG and MDA expression in liver tissue as well as TBIL, ALT, AST, γ-GT, TNF-α, IL-1β, IL-6, 8-OHdG and MDA levels in serum of model group were significantly higher than those of normal group while p-JAK2, p-STAT3, TNF-α, IL-1β, IL-6, ROS, 8-OHdG and MDA expression in liver tissue as well as TBIL, ALT, AST, γ-GT, TNF-α, IL-1β, IL-6, 8-OHdG and MDA levels in serum of PDTC group and AG490 group were significantly lower than those of model group. Conclusions PDTC can inhibit the inflammation and oxidative stress mediated by JAK2/STAT3 signaling pathway to alleviate the anti-tuberculosis drug-induced liver injury.

Incidence and outcome of anti-tuberculosis drug-induced hepatotoxicity in tuberculosis inpatients / 中华流行病学杂志

Objective Based on the medical records and follow-up records of hospitalized patients who received anti-tuberculosis therapy in the Third People' s Hospital of Zhenjiang in Jiangsu province from 2006 to 2012,we investigated the incidence and outcome of anti-tuberculosis drug induced hepatotoxicity (ATDH) and provided evidence for the prevention of ATDH.Methods According to tuberculosis patients' medical information and liver function test records,ATDH patients were diagnosed according to the criteria of International Consensus Meeting and American Thoracic Society respectively,then the related factors and outcomes were analyzed.Results A total of 1 967 hospitalized tuberculosis patients were reviewed retrospectively,in which 1 403 (71.3％) were men,1 790 (91.0％) were pulmonary tuberculosis patients,1 528 (77.8％) were patients receiving initiative treatment,979 (49.8％) were sputum smear-positive patients,and 1 297 (65.9％) had other complicated diseases.According to the criterion of International Consensus Meeting,the incidence of ATDH was 16.5％,the median time of onset was 25 days.According to the criterion of American Thoracic Society,the incidence of ATDH was 8.3％,the median time of onset was 23 days.The incidence of ATDH was significantly higher in males and HRZE therapy group (P＜0.05).Under the two liver criteria,69.5％ and 70.1％ of the patients changed primary therapy respectively after ATDH occurred.89.8％ and 88.4％ patients' liver function returned to normal range after changing or stopping therapy.Conclusion According to two liver injury criteria,the incidences of ATDH were 16.5％ and 8.3％ in hospitalized tuberculosis patients respectively,and ATDH mainly occurred in the furst month of anti-tuberculosis treatment.The monitoring of liver function should be strengthened in males and HRZE therapy group to reduce the incidence of ATDH.

Incidence and outcome of anti-tuberculosis drug-induced hepatotoxicity in tuberculosis inpatients / 中华流行病学杂志

Objective Based on the medical records and follow-up records of hospitalized patients who received anti-tuberculosis therapy in the Third People' s Hospital of Zhenjiang in Jiangsu province from 2006 to 2012,we investigated the incidence and outcome of anti-tuberculosis drug induced hepatotoxicity (ATDH) and provided evidence for the prevention of ATDH.Methods According to tuberculosis patients' medical information and liver function test records,ATDH patients were diagnosed according to the criteria of International Consensus Meeting and American Thoracic Society respectively,then the related factors and outcomes were analyzed.Results A total of 1 967 hospitalized tuberculosis patients were reviewed retrospectively,in which 1 403 (71.3％) were men,1 790 (91.0％) were pulmonary tuberculosis patients,1 528 (77.8％) were patients receiving initiative treatment,979 (49.8％) were sputum smear-positive patients,and 1 297 (65.9％) had other complicated diseases.According to the criterion of International Consensus Meeting,the incidence of ATDH was 16.5％,the median time of onset was 25 days.According to the criterion of American Thoracic Society,the incidence of ATDH was 8.3％,the median time of onset was 23 days.The incidence of ATDH was significantly higher in males and HRZE therapy group (P＜0.05).Under the two liver criteria,69.5％ and 70.1％ of the patients changed primary therapy respectively after ATDH occurred.89.8％ and 88.4％ patients' liver function returned to normal range after changing or stopping therapy.Conclusion According to two liver injury criteria,the incidences of ATDH were 16.5％ and 8.3％ in hospitalized tuberculosis patients respectively,and ATDH mainly occurred in the furst month of anti-tuberculosis treatment.The monitoring of liver function should be strengthened in males and HRZE therapy group to reduce the incidence of ATDH.

Multiplex Assay of Second-Line Anti-Tuberculosis Drugs in Dried Blood Spots Using Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry

As dried blood spots (DBSs) have various advantages over conventional venous blood sampling, some assays for detection of one or two anti-tuberculosis (TB) drugs in DBSs have been developed. However, there are no assays currently available for the simultaneous measurement of three or more anti-TB drugs in DBSs. In this study, we developed and evaluated a multiplex method for detecting nine anti-TB drugs including streptomycin, kanamycin, clarithromycin, cycloserine, moxifloxacin, levofloxacin, para-aminosalicylic acid, prothionamide, and linezolid in DBSs by using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Seventy-nine patient samples of DBS were analyzed on the UPLC-MS/MS system. All drug concentrations were determined within 4 min, and assay performance was evaluated. All drugs were clearly separated without ion suppression. Within-run and between-run precisions were 1.7-13.0% and 5.7-17.0%, respectively, at concentrations representing low and high levels for the nine drugs. Lower limits of detection and quantification were 0.06-0.6 and 0.5-5.0 µg/mL, respectively. Linearity was acceptable at five level concentrations for each drug. Correlations between drug concentrations in plasma and DBSs by using Passing-Bablock regression and Pearson's rho (ρ, 0.798-0.989) were acceptable. In conclusion, we developed a multiplex assay to measure nine second-line anti-TB drugs in DBSs successfully. This assay provided convenient and rapid drug quantification and could have applications in drug monitoring during treatment.

Characteristic of Liver Injury Induced by Simvastatin Combined with HRZ in SD Rats / 昆明医科大学学报

Objective To study the characteristics of liver injury induced by simvastatin combined with HRZ (Isoniazid,Rifampicin and Pyrazinamide) in SD rats.Methods Fifty-four 8-week-old SD rats were randomly divided into 3 groups:group A (control),group B (HRZ) and group C (simvastatin combined to HRZ),half of each group were male.We calculated the accurate dose respectively before those rats were given intragastrical administration of corresponding drugs.Six rats were killed in each group on 10th,20th and 40th day,respectively.Before this,blood was fastened from femoral of every rat that would be killed to test liver function,liver tissue slices were made in order to observe the pathological characteristic.Results Alanine amiotransferase of group C elevated in line with time and reached statistic difference on 40th day,furthermore,it was significantly higher than group A (P＜0.05).Total bilirubin and direct Bilirubin of group C were significantly higher than those of group A from the beginning to the end (P＜0.05),however,they declined rapidly on 10th day,this trend also had statistic difference (P＜0.05) At the end of this experiment,hepatic cords was disordered slightly,but swelling liver cells and vacuolar degeneration were observed,the nuleus of cell condensed.Soakage of monocytes,neutrophils,and lymphocytes occurred in the portal and lobule regions,or even spotty necrosis occasionally.Conclusion Cholestasis occurs at the early stage when simvastatin is combined with HRZ in SD rats,however,it has a rapidly degressive trend.In contrast,Alanine amiotransferase elevates,furthermore,pathological injury or even spotty necrosis can emerge in liver tissue slices.