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Objectives: The definitive diagnosis of endobronchial tuberculosis (EBTB) is challenging because the disease manifests in various non-specific ways, and acid-fast bacilli (AFB) are often undetectable by sputum smear. The objective of this study was to evaluate the efficacy of real-time PCR of bronchoscopic biopsy specimens for the diagnosis of EBTB. Methods: Real-time PCR amplification of Mycobacterium tuberculosis DNA in biopsy tissue from EBTB patients was performed prospectively. Diagnostic yields were compared for real-time PCR and for auramine O-stained sputum smears and bronchial brush smears. Whether diagnostic yield depended on bronchoscopic subtype of EBTB was also evaluated. Results: Diagnostic yields were 4.1% (3/74) for sputum smear, 39.2% (29/74) for bronchial brush smear, and 89.2% (66/74) for real-time PCR. Real-time PCR melting curve analysis showed significantly higher yields than did AFB staining of bronchial brush smears for granular and caseating EBTB (p

Theabrownins (TBs) are the characteristic functional and quality components of dark teas such as Pu'er tea and Chin-brick tea. TBs are a class of water-soluble brown polymers with multi-molecular weight distribution produced by the oxidative polymerisation of tea polyphenols during the fermentation process of dark tea, both enzymatically and non-enzymatically. TBs have been extracted and purified from dark tea all the time, but the obtained TBs contain heterogeneous components such as polysaccharides and caffeine in the bound state, which are difficult to remove. The isolation and purification process was tedious and required the use of organic solvents, which made it difficult to industrialise TBs. In this study, epigallocatechin (EGC), epigallocatechin gallate (EGCG), epigallocatechin gallate (ECG), EGC/EGCG (mass ratio 1:1), EGCG/ECG (mass ratio 1:1), EGC/ECG (mass ratio 1:1) and EGC/EGCG/ECG (mass ratio 1:1:1) as substrates and catalyzed by polyphenol oxidase (PPO) and peroxidase (POD) in turn to produce TBs, named TBs-dE-1, TBs-dE-2, TBs-dE-3, TBs-dE-4, TBs-dE-5, TBs-dE-6 and TBs-dE-7. The physicochemical properties and the antibacterial activity and mechanism of TBs-dE-1–7 were investigated. Sensory and colour difference measurements showed that all seven tea browning samples showed varying degrees of brownish hue. Zeta potential in aqueous solutions at pH 3.0–9.0 indicated that TBs-dE-1–7 was negatively charged and the potential increased with increasing pH. The characteristic absorption peaks of TBs-dE-1–7 were observed at 208 and 274 nm by UV-visible (UV-vis) scanning spectroscopy. Fourier transform infrared (FT-IR) spectra indicated that they were phenolic compounds. TBs-dE-1–7 showed significant inhibition of Escherichia coli DH5α (E. coli DH5α). TBs-dE-3 showed the strongest inhibitory effect with minimum inhibitory concentration (MIC) of 1.25 mg mL–1 and MBC of 10 mg mL–1, followed by TBs-dE-5 and TBs-dE-6. These three TBs-dEs were selected to further investigate their inhibition mechanism. The TBs-dE was found to damage the extracellular membrane of E. coli DH5α, causing leakage of contents, and increase intracellular reactive oxygen content, resulting in abnormal cell metabolism due to oxidative stress. The results of the study provide a theoretical basis for the industrial preparation and product development of TBs.

Background and aims: Idiopathic pulmonary fibrosis (IPF) is a fibrosing lung disease with unknown etiology, leading to cough and dyspnoea, which is also one of the most common sequelae affecting the quality of life of COVID-19 survivors. There is no cure for IPF patients. We aim to develop a reliable IPF animal model with quantification of fibrosis based on Micro-Computer Tomography (micro-CT) images for the new drug discovery, because different bleomycin administration routes, doses, and intervals are reported in the literature, and there is no quantitative assessment of pulmonary fibrosis based on micro-CT images in animal studies. Methods: We compared three dosages (1.25 mg/kg, 2.5 mg/kg, and 5 mg/kg) of intratracheal bleomycin administration and experiment intervals (14 and 21 days) in C57BL/6 mice by investigating survival rates, pulmonary histopathology, micro-CT, peripheral CD4+ & CD8+ cells, and cytokines. Moreover, a simple and reliable new method was developed for scoring fibrosis in live mice based on Micro-CT images by using Image J software, which transfers the dark sections in pulmonary Micro-CT images to light colors on a black background. Results: The levels of hydroxyproline, inflammation cytokine, fibrotic pathological changes, and collagen deposition in the lungs of mice were bleomycin dose-dependent and time-dependent as well as the body weight loss. Based on the above results, the mice model at 21 days after being given bleomycin at 1.25 mg/kg has optimal pulmonary fibrosis with a high survival rate and low toxicity. There is a significant decrease in the light area (gray value at 9.86 ± 0.72) in the BLM mice, indicating that a significant decrease in the alveolar air area was observed in BLM injured mice compared to normal groups (###p < 0.001), while the Pirfenidone administration increased the light area (gray value) to 21.71 ± 2.95 which is close to the value observed in the normal mice (gray value at 23.23 ± 1.66), which is consistent with the protein levels of Col1A1, and α-SMA. Notably, the standard deviations for the consecutive six images of each group indicate the precision of this developed quantitation method for the micro-CT image taken at the fifth rib of each mouse. Conclusion: Provided a quantifying method for Micro-CT images in an optimal and repeatable pulmonary fibrosis mice model for exploring novel therapeutic interventions.

Carotenoid biosynthetic pathway produces not only pigments that protect photosynthetic system against photo-oxidative damage, but also precursors of abscisic acid, the major hormone regulates stress responses. To understand the response of carotenoid biosynthetic pathway to salt stress, the expression of the genes involved in carotenoid and ABA biosynthesis were compared in cultivated tomato Solanum lycopersicon cv. Moneymaker and its relative wild genotype S. pimpinellifolium (PI365967) together with the contents of carotenoids and ABA. The results showed that 11 of the 15 genes investigated were up-regulated and four unaltered in Moneymaker after 5 h of salt stress; whereas only four genes were up-regulated, four unaltered, and seven down-regulated in PI365967 after stress. Further comparison revealed that 11 salinity-induced genes were expressed significantly lower in Moneymaker than in PI365967 under normal condition, and 8 of them were induced to similar levels after salt stress. In consistence, ABA level was doubled in Moneymaker but kept consistent in PI365967 after salt stress, though the contents of neoxanthin, violaxanthin, β-carotene, lutein, and total carotenoids were kept unchanged in both species. Since it is known that PI365967 is more tolerant to salt stress than Moneymaker, we proposed that the constitutive high level of carotenoid and ABA biosynthetic pathway under normal growth condition could be benefit to PI365967 for establishing the early response to salt stress. In addition, CrtR-b1 and CrtR-b2 that encode β-carotenoid hydroxylases were the only genes in carotenoid biosynthetic pathway that were up-regulated by salt stress in both species. The CrtR-b2 gene was cloned from both species and no essential difference was found in the encoded amino acid sequences. Transformation of CrtR-b2 to tobacco improved the seed germination under salt stress condition, indicating that the hydrolysis of β-carotenoid is the target of transcriptional regulation of the carotenoid biosynthesis in both tomato cultivar and wild relative.