Use of T-SPOT.TB for the diagnosis of unconventional pleural tuberculosis is superior to ADA in high prevalence areas: a prospective analysis of 601 cases.

BACKGROUND: Tuberculous pleural effusion (TPE) is the most common extrapulmonary manifestation and may have lasting effect on lung function. However conventional diagnostic tests for TPE register multiple limitations. This study estimates diagnostic efficacy of the interferon gamma release assay (IGRA: T-SPOT.TB) in TPE patients of different characteristics. METHODS: We performed a prospective, single-centre study including all suspected pleural effusion patients consecutively enrolled from June 2015 to October 2018. Through receiver operating characteristic (ROC) curves, technical cut-offs and the utility of T-SPOT on pleural fluid (PF) were determined and analysed. Logistic regression analysis was performed to obtain the independent risk factors for TPE, and evaluated the performance of the T-SPOT assay stratified by risk factors in comparison to ADA. RESULTS: A total of 601 individuals were consecutively recruited. The maximum spot-forming cells (SFCs) of early secretory antigenic target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10) in the PF T-SPOT assay had the best diagnostic efficiency in our study, which was equal to ADA (0.885 vs 0.887, P = 0.957) and superior to peripheral blood (PB), with a sensitivity of 83.0% and a specificity of 83.1% (The cut-off value was 466 SFCs/106 mononuclear cells). Among the TPE patients with low ADA (< 40 IU/L), the sensitivity and specificity of PF T-SPOT were still 87.9 and 90.5%, respectively. The utility of ADA was negatively related to increasing age, but the PF T-SPOT test had a steady performance at all ages. Age (< 45 yrs.; odds ratio (OR) = 5.61, 95% confidence interval (CI) 3.59-8.78; P < 0.001), gender (male; OR = 2.68, 95% CI 1.75-2.88; P < 0.001) and body mass index (BMI) (< 22; OR = 1.93, 95% CI 1.30-2.88; P = 0.001) were independently associated with the risk of TB by multivariate logistic regression analysis. Notably, when stratified by risk factor, the sensitivity of PF T-SPOT was superior to the sensitivity for ADA (76.5% vs. 23.5%, P = 0.016) and had noninferior specificity (84.4% vs. 96.9%, P = 0.370). CONCLUSIONS: In conclusion, the PF T-SPOT assay can effectively discriminate TPE patients whose ADA is lower than 40 IU/L and is superior to ADA in unconventional TPE patients (age ≥ 45 yrs., female or BMI ≥ 22). The PF T-SPOT assay is an excellent choice to supplement ADA to diagnose TPE.

Assessment of Interferon-Gamma Release Assay in Patients with Non-Tuberculous Mycobacteria Pulmonary Disease.

BACKGROUND: Interferon-gamma release assay (T-SPOT.TB) has the theoretical possibility of discriminating TB from most non-tuberculous mycobacteria (NTM) infections, but there are limited reports on the use of T-SPOT.TB for diseases due to NTM in high TB burden country. The aim of the present study was to assess the utility of T-SPOT.TB in patients with NTM pulmonary disease. METHODS: Clinical parameters and laboratory characteristics of patients with NTM pulmonary disease between July 2011 and Jan 2017 were investigated retrospectively and comprehensively reviewed. RESULTS: A total of 127 patients with NTM pulmonary disease were retrospectively reviewed. Seven NTM species were isolated from 115 patients, and the most common species were M. intracellulare (48.7%, 56/115) and M. abscessus (34.8%, 40/115). NTM isolates were mainly prevalent in people aged 50 years or older (73.0%). The overall positive rate of T-SPOT.TB test was 29.6% (24/81). In patients infected with NTM sharing the RD1 region of Mycobacterium tuberculosis (M. TB), 50% (3/6) were positive in the T-SPOT.TB test, whereas 28.0% (21/75) was positive in the group with NTM not sharing the RD1 region of M. TB. No significant difference was detected in the positive rate of T-SPOT.TB between definite (28.3%, 15/53) and probable disease (32.1%, 9/28). CONCLUSIONS: Our data indicated a relatively high positive rate of T-SPOT.TB test in patients infected with NTM not sharing the RD1 region of M. TB. Thus, T-SPOT.TB test displays a limited ability in differentiating TB infection from NTM disease in a high TB burden country.

Transmitted Extended-Spectrum Extensively Drug-Resistant Tuberculosis in Beijing, China, with Discordant Whole-Genome Sequencing Analysis Results.

Drug resistance to tuberculosis remains a major public health threat. Here, we report two cases of extended-spectrum extensively drug-resistant (XXDR) tuberculosis showing resistance to most first- and second-line agents. The results of a correlation of whole-genome sequencing (WGS) and phenotypic testing were discordant, suggesting that overreliance on WGS may miss clinically relevant resistance in extensively drug-resistant disease.

ALDH2*2 Allele is a Negative Risk Factor for Cerebral Infarction in Chinese Women.

Unlike its reported role in the cardiovascular diseases, little information is available for mitochondrial aldehyde dehydrogenase 2 (ALDH2) in the cerebrovascular function. We investigated the different effects of ALDH2 genotypes on the risk of cerebral infarction between the genders, because different genders had different smoking and/or dinking status which are also risk factors for cerebral infarction. 247 healthy Chinese Han people (controls, group 1), 287 Chinese Han male patients with cerebral infarction (group 2), and 82 Chinese Han female patients with cerebral infarction (group 3) were involved in this study. The frequencies of the ALDH2*2 allele in group 3 were significantly higher than those in other groups (with P = 0.001 and P = 0.002, respectively). The difference of ALDH2*2 allele frequency between group 1 and group 2 was not significant (P = 0.652). After adjustment for smoking and drinking status, the male patients without smoking or drinking status (group 4) had higher ALDH2*2 allele frequency than group 1, but the difference was still not significant (P = 0.139). Thus, we conclude that ALDH2*2 allele may be a significant negative risk factor for cerebral infarction in Chinese women [odds ratio (OR) = 2.207, 95% CI 1.416-3.439]. But for Chinese male patients, the negative effects of ALDH2*2 allele on cerebral infarction which might be concealed by other risk factors were not significant.

[Jaridonin induces apoptosis in human esophageal cancer cells by depleting GSH and inducing DNA damage].

OBJECTIVE: The aim of this study was to explore the molecular mechanism of apoptosis in esophageal cancer cells induced by Isodon rubescens. METHODS: The DNA-damage effect of Jaridonin was detected by single cell gel electrophoresis (SCGE). The p53 protein was determined by Western blot. GSH assay kit was employed to determine the GSH content in human esophageal cancer EC-1 cells. Intracellular levels of hydrogen peroxide (H2O2) or superoxide (O(2).-) were determined using the redox-sensitive probes 2', 7'-dichlorodihydrofluorescein diacetate (DCF) or dihydroethidium (DHE), and the fluorescence signal was assayed by fluorescence microscopy and by flow cytometry. RESULTS: Jaridonin induced DNA damage in EC-1 cells remarkably. The olive tail moments (OTM) of control and 20, 40 µmol/L Jaridonin were 3.2, 45.2 and 89.0, respectively. Compared with the control, the differences were significant (P < 0.01 for both). Jaridonin resulted in extensive p53 up-regulation in the EC-1 cells. More importantly, the p53 up-regulation occurred as early as 2 h after Jaridonin incubation, and in a time-dependent manner (P < 0.05). p53 siRNA transfection inhibited apoptosis in the EC-1 cells, and the Jaridonin-induced apoptosis rate was reduced from 38.5% to 8.8%. Intracellular level of H2O2 was increased by Jaridonin, whereas the level of O(2).- was barely changed. The GSH content in EC-1 cells was reduced from (10.3 ± 1.6) nmol/mg protein to (4.6 ± 2.1) nmol/mg protein after 20 µmol/L Jaridonin incubation for 8 h, and it was further reduced with the increase of Jaridonin concentration. Jaridonin induced DNA damage, H2O2 accumulation and apoptosis were significantly attenuated in the presence of GSH, but Jaridonin showed little effect on normal human liver L-02 cells. CONCLUSIONS: Jaridonin selectively induces apoptosis in esophageal cancer EC-1 cells through H2O2-mediated DNA damage by depleting GSH.

Individuals having variant genotypes of cytochrome P450 2C19 are at increased risk of developing primary liver cancer in Han populations, without infection with the hepatitis virus.

Recently, many researchers have reported that the genetic polymorphisms of CYP2C19 may account for the interpatient variability of the clinical course in cancers including primary liver cancer (PLC). Besides the genetic polymorphisms of CYP2C19, hepatitis viruses (HV, including HAV, HBV, HCV, HDV, HEV, especially HBV and/or HCV) also account for the interpatient variability of the clinical course in PLC. This research covered the above two factors and divided the patients with PLC into two groups (one group with HBV infection and another without any HV infection) to find out whether the genetic polymorphisms of CYP2C19 have different effects in the progressing of PLC in different groups of patients. Eight hundred sixty-four cancer-free Han people (controls, named group 1), 207 Han PLC patients with HBV infection (group 2), and 55 Han PLC patients without any HV infection (group 3) were involved in this study. A wild-type allele (CYP2C19*1) and two mutated alleles (CYP2C19*2 and CYP2C19*3) were identified. The frequencies of the mutant alleles and genotypes were then compared with each other. The frequencies of the homozygous and heterozygous variant genotypes (*2/*2, *2/*3, *3/*3) in group 3 (25.5 %) were significantly higher than those in other groups (11.9 % in group 1 and 13.5 % in group 2, P = 0.014, 95 % confidence interval (CI)). The differences were statistically significant between group 1 and group 3 (P = 0.004, 95 % CI), but they were not statistically significant between group 1 and group 2 (P = 0.527, 95 % CI). Thus, we conclude that people which were not infected with HV but with the homozygous or heterozygous variant genotypes (*2/*2, *2/*3, *3/*3) of CYP2C19 may have higher possibilities of getting PLC than people with other allelic genotypes (*1/*1, *1/*2, *1/*3) (odds ratio (OR) = 2.523, 95 % CI = 1.329 ~ 4.788). However, in patients with HBV infection, the genetic polymorphisms of CYP2C19 did not seem to be an important factor in the risk of developing PLC (OR = 1.156, 95 % CI = 0.738 ~ 1.810).

[Macrolide and linezolid susceptibility testing for Mycobacterium intracellulare isolates].

OBJECTIVE: To investigate the susceptibility of Mycobacterium intracellulare isolates to clarithromycin, azithromycin and linezolid, and therefore to explore the possibility of using these drugs to treat Mycobacterium intracellulare diseases. METHODS: The minimal inhibitory concentrations (MICs) of the 3 antibiotics against 76 Mycobacterium intracellulare isolates were determined by using microplate alamar blue assay (MABA). RESULTS: The MIC90 of clarithromycin against Mycobacterium intracellulare isolates was 2 mg/L. The proportion of susceptible, intermediate and resistant isolates to clarithromycin was 93.4% (71/76), 0.0% (0/76) and 6.6% (5/76), respectively. The MIC90 of azithromycin against the isolates was 32 mg/L. The proportion of susceptible, intermediate and resistant isolates to azithromycin was 94.7% (72/76), 0.0% (0/76) and 5.3% (4/76), respectively. The MIC90 of linezolid was 64 mg/L. The proportion of susceptible, intermediate and resistant isolates to linezolid was 32.9% (25/76), 22.4% (17/76) and 44.7% (34/76), respectively. Among the 5 isolates resistant to clarithromycin, 4 were resistant to azithromycin, and 2 were resistant to linezolid. CONCLUSIONS: Most of the Mycobacterium intracellulare isolates were sensitive to clarithromycin. High cross-resistance between clarithromycin and azithromycin was present, but some clarithromycin-resistant isolates were sensitive to linezolid.

[The analysis of species identification and susceptibility testing of Mycobacterium abscessus].

OBJECTIVES: To evaluate the method of differentiating Mycobacterium abscessus subsp. abscessus (subsp. M. abscessus) from Mycobacterium abscessus subsp. massiliense (subsp. M. massiliense), and to investigate the activity of different antibiotics in vitro. METHODS: Sixty Mycobacterium abscessus (M. abscessus) isolates previously identified by using 16S rRNA sequence, were identified by comparative sequence analysis of rpoB and hsp65, and were divided into subsp. M. abscessus and subsp. M. massiliense. Two subspecies' resistant proportions were compared by chi-square test. Finally the relationship between clarithromycin resistance and erm(41) was analyzed. RESULTS: Of all the 60 M. abscessus isolates, 65% (39/60) belonged to subsp. M. abscessus, 35% (21/60) belonged to subsp. M. massiliense. 97% (38/39) subsp. M. abscessus and 95% (20/21) subsp. M. massiliense were susceptible to amikacin. 92% (36/39) subsp. M. abscessus and 95% (20/21) subsp. M. massiliense were susceptible to azithromycin. 74% (29/39) subsp. M. abscessus and 67% (14/21) subsp. M. massiliense were susceptible to imipenem. 46% (18/39) subsp. M. abscessus and 76% (16/21) subsp. M. massiliense were moderately susceptible to cefoxitin. 82% (32/39) subsp. M. abscessus were resistant to clarithromycin, and 95% (20/21) subsp. M. massiliense were susceptible to clarithromycin. Of all the 28 subsp. M. abscessus isolates which were sequenced suscessfully, 23 isolates were resistant to clarithromycin, and 22 isolates in them had T28 in erm(41), and the rest one had C28 in erm(41). However, all the 5 subsp. M. abscessus isolates which were susceptible to clarithromycin had C28 in erm(41). CONCLUSIONS: M. abscessus can be divided into subsp. M. abscessus and subsp. M. massiliense by using rpoB and hsp65. Amikacin, azithromycin and imipenem showed excellent inhibition activity against M. abscessus in vitro. Cefoxitin also showed a good inhibition activity. Clarithromycin had a poor inhibition activity against subsp. M. abscessus, but a good inhibition activity against subsp. M. massiliense. Erm(41) was related to clarithromycin resistance.

Efficacy of clofazimine-containing regimens for treatment of Mycobacterium avium complex-pulmonary disease in patients unsuitable for standard treatment regimen.

OBJECTIVES: Patients with Mycobacterium avium complex-pulmonary disease (MAC-PD) can exhibit contraindications in applying the recommended treatment regimens by the guidelines. Clofazimine (CFZ) is considered a promising drug for MAC-PD treatment and is frequently included in alternative regimens; however, its efficacy remains unclear. METHODS: MAC-PD patients, unsuitable for standard regimens, were enrolled continuously in a prospective study at Beijing Chest Hospital. The treatment response of the CFZ-containing regimen was monitored. RESULTS: Fifty patients were enrolled in the initial treatment, and 25 patients had a history of anti-TB treatment. Nodular bronchiectasis was observed in 34 patients, while 8 patients exhibited fibrocavitary changes. Additionally, eight patients displayed a combination of both patterns. In a multivariate analysis, MAC-PD patients with CFZ MIC < 0.25 mg/L were significantly associated with culture conversion [OR 8.415, 95% CI (1.983-35.705); P = 0.004]. Among patients who had previous TB treatment history, patients with CFZ MIC < 0.25 mg/L had a higher chance of acquiring culture conversion outcomes [(OR 7.737, 95% CI 1.032-57.989); P = 0.046]. In contrast, among patients with no previous TB treatment history, the RIF-containing regimen had a higher chance of acquiring culture conversion outcomes [(OR 11.038, 95%CI 1.008-120.888); P = 0.049]. CONCLUSION: MAC-PD patients unsuitable for standard regimens could benefit from a CFZ-containing regimen, especially for patients with previous TB treatment history and baseline CFZ MIC values lower than 0.25 mg/L.

A comparative study of MassARRAY and GeneXpert assay in detecting rifampicin resistance in tuberculosis patients' clinical specimens.

Objectives: Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has emerged as a potent tool for detecting drug resistance in tuberculosis (TB); however, concerns about its reliability have been raised. In this study, we assessed the reliability of MassARRAY (Sequenom, Inc.), which is a MALDI-TOF MS-based method, by comparing it to the well-established GeneXpert assay (Cepheid) as a reference method. Methods: A retrospective study was conducted using laboratory data retrieved from Henan Chest Hospital (Zhengzhou, China). To ensure a rigorous evaluation, we adopted a comprehensive assessment approach by integrating multiple outcomes of the Xpert assay across various specimen types. Results: Among the 170 enrolled TB cases, MassARRAY demonstrated significantly higher sensitivity (85.88%, 146 of 170) compared to the Xpert assay (76.62%, 118 of 154) in TB diagnosis (p < 0.05). The concordance in detecting rifampicin resistance between MassARRAY and the combined outcomes of the Xpert assay was 90%, while it was 97.37% (37 of 38) among smear-positive cases and 89.06% (57 of 64) among culture-positive cases. When compared to the phenotypic susceptibility outcomes of the 12 included drugs, consistency rates of 81.8 to 93.9% were obtained, with 87.9% for multiple drug resistance (MDR) identification. Conclusion: MassARRAY demonstrates high reliability in detecting rifampicin resistance, and these findings may offer a reasonable basis for extrapolation to other drugs included in the test panel.

Bedaquiline susceptibility testing of Mycobacterium abscessus complex and Mycobacterium avium complex: A meta-analysis study.

OBJECTIVE: This study aims to estimate the overall in vitro activity of bedaquiline (BDQ) against clinical isolates of Mycobacterium abscessus complex (MABS) and M. avium complex (MAC), considering BDQ as a repurposed drug for non-tuberculous mycobacteria (NTM) infections. METHODS: We conducted a systematic review of publications in PubMed/ MEDLINE, Web of Science, and Embase up to 15 April 2023. Studies were included if they followed the Clinical and Laboratory Standards Institute (CLSI) criteria for drug susceptibility testing (DST). Using a random effects model, we assessed the overall in vitro BDQ resistance rate in clinical isolates of MABS and MAC. Sources of heterogeneity were analysed using Cochran's Q and the I2 statistic. All analyses were performed using CMA V3.0. RESULTS: A total of 24 publications (19 reports for MABS and 11 for MAC) were included. Using 1 µg/mL and 2 µg/mL as the breakpoint for BDQ resistance, the pooled rates of in vitro BDQ resistance in clinical isolates of MABS were found to be 1.8% (95% confidence interval [CI], 0.7-4.6%) and 1.7% (95% CI, 0.6-4.4%), respectively. In the case of MAC, the pooled rates were 1.7% (95% CI, 0.4-6.9%) and 1.6% (95% CI, 0.4-6.8%) for 1 µg/mL and 2 µg/mL, respectively. CONCLUSION: This study reports the prevalence of BDQ resistance in clinical isolates of MABS and MAC. The findings suggest that BDQ holds potential as a repurposed drug for treating MABS and MAC infections.

Acute ischemic stroke in tuberculous meningitis.

Background: The underlying mechanism for stroke in patients with tuberculous meningitis (TBM) remains unclear. This study aimed to investigate the predictors of acute ischemic stroke (AIS) in TBM and whether AIS mediates the relationship between inflammation markers and functional disability. Methods: TBM patients admitted to five hospitals between January 2011 and December 2021 were consecutively observed. Generalized linear mixed model and subgroup analyses were performed to investigate predictors of AIS in patients with and without vascular risk factors (VAFs). Mediation analyses were performed to explore the potential causal chain in which AIS may mediate the relationship between neuroimaging markers of inflammation and 90-day functional outcomes. Results: A total of 1,353 patients with TBM were included. The percentage rate of AIS within 30 days after admission was 20.4 (95% CI, 18.2-22.6). A multivariate analysis suggested that age ≥35 years (OR = 1.49; 95% CI, 1.06-2.09; P = 0.019), hypertension (OR = 3.56; 95% CI, 2.42-5.24; P < 0.001), diabetes (OR = 1.78; 95% CI, 1.11-2.86; P = 0.016), smoking (OR = 2.88; 95% CI, 1.68-4.95; P < 0.001), definite TBM (OR = 0.19; 95% CI, 0.06-0.42; P < 0.001), disease severity (OR = 2.11; 95% CI, 1.50-2.90; P = 0.056), meningeal enhancement (OR = 1.66; 95% CI, 1.19-2.31; P = 0.002), and hydrocephalus (OR = 2.98; 95% CI, 1.98-4.49; P < 0.001) were associated with AIS. Subgroup analyses indicated that disease severity (P for interaction = 0.003), tuberculoma (P for interaction = 0.008), and meningeal enhancement (P for interaction < 0.001) were significantly different in patients with and without VAFs. Mediation analyses revealed that the proportion of the association between neuroimaging markers of inflammation and functional disability mediated by AIS was 16.98% (95% CI, 7.82-35.12) for meningeal enhancement and 3.39% (95% CI, 1.22-6.91) for hydrocephalus. Conclusion: Neuroimaging markers of inflammation were predictors of AIS in TBM patients. AIS mediates < 20% of the association between inflammation and the functional outcome at 90 days. More attention should be paid to clinical therapies targeting inflammation and hydrocephalus to directly improve functional outcomes.

[Clinical analysis of protionamide and para-aminosalicylic acid induced hepatotoxicity in 129 cases].

OBJECTIVE: To investigate drug-induced liver injury (DILI) in tuberculosis (TB) patients treated with protionamide (Pto) and (or) para-aminosalicylic acid (PAS), and therefore to provide data for using second-line anti-tuberculosis drugs and risk prediction of liver damage. METHODS: A retrospective analysis was performed for TB patients treated with regimens containing Pto and (or) PAS in Beijing Chest Hospital during Jan. 2008 to Jan. 2013. Cases with DILI were identified, and associated factors including patients' age and gender, time of onset, severity, clinical manifestations and prognosis of DILI were analyzed. The 2 groups were compared with χ(2) test. P < 0.05 was considered to be significant. RESULTS: A total of 1714 cases were admitted, among whom 226 experienced liver damage during treatment, of which 97 cases were excluded because of underlying alcoholic liver disease, viral hepatitis B and C. Finally, 129 cases were diagnosed as having DILI, resulting in an overall incidence of 7.5% (129/1714), being 9.2% (59/641) in females, and 6.5% (70/1073) in males (χ(2) = 4.143, P < 0.05). DILI in most patients occurred between 1 week to 2 months, with 30.2% (39/129) within 2-4 weeks. 47.3% (61/129) of the patients showed no obvious clinical symptoms of hepatotoxicity. Among different regimens, combination of Pto, PAS and PZA resulted in the highest rate of DILI (20.7%, 19/92), while the rate was 9.8% (8/82) for the combination of Pto and PZA, P < 0.05. CONCLUSIONS: DILI caused by Pto and PAS should be taken into account, especially in female patients and for multi-drug combination therapy. Liver function should be monitored even in patients without related clinical manifestations for early identification and treatment, and therefore avoiding severe liver damage.

[Mycobacterium abscessus group lung disease: case reports and review of the literature].

OBJECTIVE: To analyze the clinical manifestations and efficacy of a combination antibiotic therapy including cefoxitin for Mycobacterium abscessus (M.abscessus) group lung disease. METHODS: We retrospectively analyzed the clinical manifestations of 16 patients with M.abscessus group lung disease, and the responses of 5 cases treated with whole-course clarithromycin and moxifloxacin, initially intensified with intravenous amikacin and cefoxitin therapy for the first 12 weeks. RESULTS: Radiological study showed that 14 patients with M.abscessus group pulmonary disease were classified as nodular bronchiectasis form, and 1 patient as upper lobe cavity form and 1 patient was unclassifiable. The radiological characteristics of M.abscessus group pulmonary disease included multiple micronodules (14/16), bronchiectasis (14/16), tree in bud sign (13/16), cavity (5/16), consolidation (5/16), nodules (5/16), and collapse of lung (3/16). Five cases were treated with a combination antibiotic therapy including cefoxitin. After 3 months treatment for the initial phase, 2 of them got improvement in symptoms, CT manifestations and sputum conversion. Two of them improved in symptoms and CT manifestations, but not in sputum conversion. One case showed no improvement in the initial phase, and continuation therapy also failed to improve symptoms, CT abnormalities or sputum conversion. CONCLUSIONS: Nodular bronchiectasis is the main manifestation of M.abscessus group lung disease. The main imaging characteristics included multiple micronodules, bronchiectasis and tree in bud sign. A therapeutic regimen including cefoxitin may be moderately effective in treating M.abscessus group lung disease.

Systemic immune dysregulation in severe tuberculosis patients revealed by a single-cell transcriptome atlas.

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb) infection, is currently the deadliest infectious disease in human that can evolve to severe forms. A comprehensive immune landscape for Mtb infection is critical for achieving TB cure, especially for severe TB patients. We performed single-cell RNA transcriptome and T-cell/B-cell receptor (TCR/BCR) sequencing of 213,358 cells from 27 samples, including 6 healthy donors and 21 active TB patients with varying severity (6 mild, 6 moderate and 9 severe cases). Two published profiles of latent TB infection were integrated for the analysis. We observed an obviously elevated proportion of inflammatory immune cells (e.g., monocytes), as well as a markedly decreased abundance of various lymphocytes (e.g., NK and γδT cells) in severe patients, revealing that lymphopenia might be a prominent feature of severe disease. Further analyses indicated that significant activation of cell apoptosis pathways, including perforin/granzyme-, TNF-, FAS- and XAF1-induced apoptosis, as well as cell migration pathways might confer this reduction. The immune landscape in severe patients was characterized by widespread immune exhaustion in Th1, CD8+T and NK cells as well as high cytotoxic state in CD8+T and NK cells. We also discovered that myeloid cells in severe TB patients may involve in the immune paralysis. Systemic upregulation of S100A12 and TNFSF13B, mainly by monocytes in the peripheral blood, may contribute to the inflammatory cytokine storms in severe patients. Our data offered a rich resource for understanding of TB immunopathogenesis and designing effective therapeutic strategies for TB, especially for severe patients.

[Comparison of clinical manifestations between Mycobacterium avium-intracellulare complex and Mycobacterium abscessus pulmonary diseases].

OBJECTIVE: To compare the clinical manifestations of nontuberculous mycobacterial (NTM) pulmonary diseases caused by Mycobacterium avium-intracellulare complex (MAC) and Mycobacterium abscessus. METHODS: The clinical manifestations of 18 patients with MAC and 9 patients with Mycobacterium abscessus pulmonary diseases diagnosed from 2010 to 2011 were reviewed. RESULTS: There were no significant differences in the gender, age, body mass index, predisposed diseases, symptoms and positive sputum acid-fast bacillus between MAC and Mycobacterium abscessus groups. Upper lobe cavities were more frequently observed in the MAC group (13/18), whereas nodular bronchiectatic changes were more frequent in the Mycobacterium abscessus group (3/9). Compared with MAC pulmonary diseases, several imaging characteristics were more common in the Mycobacterium abscessus group, including bilateral micro nodules (Mycobacterium abscessus group 8/9 vs MAC group 7/18), tree-in-bud sign (Mycobacterium abscessus group 7/9 vs MAC group 6/18) and multiple bronchiectasis (Mycobacterium abscessus group 8/9 vs MAC group 5/18). CONCLUSIONS: There was considerable overlap in clinical characteristics of MAC and Mycobacterium abscessus pulmonary diseases. However, bilateral micro nodules, tree-in-bud sign and multiple bronchiectasis were more frequently seen in Mycobacterium abscessus than in MAC pulmonary diseases, while upper lobe cavities were more frequently seen in MAC than in Mycobacterium abscessus pulmonary diseases.

Lysosome-Targeting Fluorescence Sensor for Sequential Detection and Imaging of Cu2+ and Homocysteine in Living Cells.

A conjugated polymer-based fluorescence sensor, namely, PTNPy, was constructed on the basis of a polythiophene scaffold coupled with dimethylpyridylamine (DPA) groups in side chains for the consecutive detection and quantification of Cu2+ and Hcy in a perfect aqueous medium. A dramatic fluorescence quenching of PTNPy by the addition of Cu2+ was observed in Tris-HCl buffer solution (2 mM, pH 7.4), demonstrating a quick (<1 min) and highly selective response to Cu2+ with a low limit of detection of 6.79 nM. Subsequently, the Cu2+-quenched fluorescence of PTNPy can be completely recovered by homocysteine (Hcy), showing excellent selectivity to Hcy over other competitive species such as cysteine and glutathione. Thanks to the low cytotoxicity and lysosomal targeting ability of PTNPy, it was further applied as an optical sensor for the sequential imaging of Cu2+ and Hcy in HeLa cells. More importantly, Hcy concentration was linearly related to the fluorescence intensity of PTNPy in living cells, demonstrating huge potential for real-time monitoring the fluctuation of Hcy levels in living cells.

Novel oxazolidinones harbor potent in vitro activity against the clinical isolates of multidrug-resistant Mycobacterium tuberculosis in China.

Objective: To investigate the in vitro activities of five oxazolidinones in parallel against the reference strains of different mycobacterial species and clinical isolates of Mycobacterium tuberculosis (Mtb), and shed light on the differences in the efficacy of these homolog drugs. Materials and methods: The minimum inhibitory concentrations (MICs) of linezolid, tedizolid, sutezolid, delpazolid, and contezolid against 16 mycobacterial reference strains and 69 M. tuberculosis clinical isolates, including 17 drug-susceptible isolates and 52 multidrug-resistant (MDR) isolates, were determined by microplate alamarBlue assay (MABA). The intracellular killing activities of contezolid and linezolid against Mtb H37Rv were compared. In addition, mutations in the linezolid resistance-related genes (rplC, rplD, and 23S rRNA) of the Mtb clinical isolates were also analyzed. Results: Tedizolid exhibited the strongest inhibitory activities against the reference strains of both rapidly growing mycobacteria (RGM) and slowly growing mycobacteria (SGM), among the tested oxazolidinones. In contrast, sutezolid only manifested potent activity against reference strains of SGM. Linezolid, delpazolid, and contezolid were less active against the non-tuberculous mycobacterial references. For the Mtb clinical isolates, the antimicrobial action was ranked as: sutezolid > tedizolid > contezolid and linezolid > delpazolid, whereas no difference between drug-sensitive and multiple drug-resistant isolates was observed. Notably, contezolid demonstrated obviously superior intracellular antimicrobial activity than linezolid. Few strains harbored mutations in rrl gene or rplD genes, although these strains had drug susceptible profiles to linezolid. Conclusion: Different oxazolidinones can have discrepant antimicrobial activity against different mycobacterial species, or have different manifestations out of cell or in cell. Understanding these differences would be helpful in choosing the appropriate drug in clinical practice.

An ethanethioate functionalized polythiophene as an optical probe for sensitive and fast detection of water content in organic solvents.

A new polythiophene-based optical probe, namely PTS, was designed and prepared for detection and quantification of the water present in organic solvents. PTS exhibited sensitive and fast absorption and fluorescence signaling response to the changes of water content in tetrahydrofuran (THF), N,N-dimethylformamide (DMF) and N,N-dimethylacetamide (DMAc) due to the water-induced interpolymer-stacking aggregation as demonstrated by dynamic light scattering (DLS) analysis. The fluorescence intensity of PTS at 550 nm linearly reduced as a function of the water content in detection ranges of 0-30% (v/v) in THF, 0-10% in DMF and 0-10% in DMAc with the limit of detection (LOD) for water being 0.034% (v/v) in THF, 0.013% (v/v) in DMF, and 0.014% (v/v) in DMAc, respectively. Additionally, PTS-incorporated test paper was fabricated to successfully achieve naked-eye detection of water in DMF and DMAc. PTS was further applied to estimate the water content in real samples, convincingly demonstrating that our method was comparable with the standard Karl Fischer titration.

[Susceptibility test of the Mycobacterium avium complex to sixteen anti-infective agents].

OBJECTIVE: To compare the in vitro activities of 13 anti-infective agents and 3 new quinolones (sitafloxacin, gatifloxacin and moxifloxacin) against Mycobacterium avium complex (MAC) isolates, and therefore to explore the possibility of using these quinolones to treat MAC diseases. METHODS: The minimal inhibitory concentration (MIC) of the above 16 anti-infective agents, including sitafloxacin, gatifloxacin and moxifloxacin against MAC isolates was determined by using agar dilution methods, and then the MIC90s of the different anti-infective agents were compared. RESULTS: The MICs of M. avium isolates showed a wider range and was less sensitive to most of the anti-infective agents as compared with M. intracellulare isolates. The MIC90s of clarithromycin against M. avium and M. intracellulare isolates were 32 mg/L and 16 mg/L, respectively, which were the lowest among 4 macrolide compounds. The MIC90 of rifalazil were 0.5 mg/L and 0.25 mg/L, respectively, which were the lowest among 4 rifamycin compounds. The MIC90 of sitafloxacin against M. avium and M. intracellulare isolates were both 4 mg/L, which were the lowest among 5 quinolones. For gatifloxacin and moxifloxacin, the MIC90 against M. avium and M. intracellulare isolates were both 8 mg/L. Two clarithromycin-sensitive strains (MIC=0.5 mg/L) showed a similar MIC of the lower limit for other compounds. Three clarithromycin-insensitive strains (MIC=64 mg/L) showed a similar MIC of the upper limit for other compounds except quinolones. CONCLUSION: Rifalazil, sitafloxacin, gatifloxacin and moxifloxacin showed acceptable in vitro activities against MAC isolates.