Evolution of triclosan resistance modulates bacterial permissiveness to multidrug resistance plasmids and phages

The horizontal transfer of plasmids has been recognized as one of the key drivers for the worldwide spread of antimicrobial resistance (AMR) across bacterial pathogens. However, knowledge remain limited about the contribution made by environmental stress on the evolution of bacterial AMR by modulating horizontal acquisition of AMR plasmids and other mobile genetic elements. Here we combined experimental evolution, whole genome sequencing, reverse genetic engineering, and transcriptomics to examine if the evolution of chromosomal AMR to triclosan (TCS) disinfectant has correlated effects on modulating bacterial pathogen (Klebsiella pneumoniae) permissiveness to AMR plasmids and phage susceptibility. Herein, we show that TCS exposure increases the evolvability of K. pneumoniae to evolve TCS-resistant mutants (TRMs) by acquiring mutations and altered expression of several genes previously associated with TCS and antibiotic resistance. Notably, nsrR deletion increases conjugation permissiveness of K. pneumoniae to four AMR plasmids, and enhances susceptibility to various Klebsiella-specific phages through the downregulation of several bacterial defense systems and changes in membrane potential with altered reactive oxygen species response. Our findings suggest that unrestricted use of TCS disinfectant imposes a dual impact on bacterial antibiotic resistance by augmenting both chromosomally and horizontally acquired AMR mechanisms

The anti-hepatic fibrosis effects of chlorogenic acid extracted from Artemisia Capillaris Herba on CCl4-induced mice via regulating TGF-β1/smad3 pathway

Introduction: Artemisia Capillaris Herba, a famous traditional Chinese medicine, is effective for the treatment of hepatic fibrosis(HF) in clinical applications. Research has confirmed that chlorogenic acid (CA), an organic acid compound was extracted from Artemisia Capillaris Herba, could reduce the hepatocyte injury induced by HF, however, its mechanism of anti-HF is still unclear, and we investigated whether CA could help treating HF mice. Methods: In this study, we evaluated the therapeutic effect of CA on HF mice induced by CCl4, which was extracted from Artemisia Capillaris Herba and identified by 1H NMR and 13C NMR spectroscopy. Seventy two NIH mice were divided into following groups: normal group, model group, low, medium and high dose of CA groups (7.5, 15, 30 mg/kg) and colchicine (Colc)-positive control group (0.2 mg/kg). All mice were injected 40% CCl4 for 8 weeks with a 24 h interval except normal mice. Each drug group and Colc group were given intragastric administration for 40 days while modeling. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), collage IV (Col-IV), hyaluronic acid (HA), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), procollagen typeⅢ (PC-Ⅲ), malondialdehyde (MDA) and laminin (LN) levels were detected by ELISA, samd3 and TGF-β1 were examined by immunohistochemistry and western blotting and the liver and kidney tissues were observed by HE. Results: At the end of administrations, the body weight of mice was decreased and the levels of ALT, AST, Col-IV, HA, IL-6, TNF-α, LN, PC-III, and MDA were increased in the HF modle mice compared with that of normal mice. Compared with the HF mice only, treatment with CA significantly decreased the levels of ALT, AST, Col-IV, HA, IL-6, TNF-α, LN, PC-III, and MDA. The HE staining results showed that the hepatic and nephritic injury were significantly alleviated after CA treatment. And the smad3 and TGF-β1 expression were inhibited in the CA-treated mice in comparison with the model mice. Conclusion: Conclusively, CA treatment could attenuate HF through the regulation of TGF-β1/smad3 pathway, suggesting that CA may be an effective component of Artemisia Capillaris Herba in the treatment of HF

Chemical constituents from the rhizome of Acorus tatarinowii Schott and their anti-inflammatory activities in LPS induced PC12 cells

Introduction: Acorus tatarinowii Schott is a traditional Chinese medicine that aromatizes dampness, opens the body and pacifies the mind, and is used medicinally as the dried rhizome of Acorus tatarinowii, family Tenaxaceae. Research has confirmed that it has a strong anti-inflammatory activity, however, its mechanism of anti-inflammatory is still unclear, and we continue to investigate its chemical composition and anti-inflammatory mechanism. Methods: The separation and purification of the EtOH extract was carried out by preparative thin layer chromatography and column chromatography. The structures of the isolated compounds were identified by NMR spectroscopy and comparing their spectral data with those previously reported in literature. CCK-8 method was used to evaluate the anti-inflammatory activity of all compounds. The MDA, NO, SOD, IL-10, IL-1β, IL-6 and TNF-α levels were analyzed using ELISA kits. TNF-α and Beclin-1 expression were examined by western blot and immunofluorescence. Results: Ten compounds, named cinnamic acid(1), sinapic acid(2), isoferulic acid(3), α-asarone(4), ferulic acid(5), arbutin (6), caffeic acid (7), 3-(3-hydroxyl phenyl)propanol(8), ethyl caffeate(9) and β-asarone(10) were isolated from the rhizome of Acorus tatarinowii Schott. Their structures were elucidated on the basis of extensive spectroscopic analyses, as well as by comparison with literature data. The anti-inflammatory effects of the all compounds were evaluated in lipopolysaccharide(LPS)-induced PC12 cell models in our study. The results of activity test showed that compounds 3 and 6 had good inhibitory effects on 5 μg·mL−1 LPS-induced PC12 cells. The IC50 of compounds 3 and 6 against PC12 cells were 27.41 ± 0.09 µmol·L−1 and 30.19 ± 0.10 µmol·L−1, respectively. Moreover, compounds 3 and 6 showed a significant protective effect on inflammation by increasing the IL-10 and SOD levels, and reducing the TNF-α, IL-6, IL-1β, MDA and NO levels. The most important point is that their anti-inflammatory effect is related to the autophagy factor Beclin-1. Conclusion: These findings imply that compounds 3 and 6 exerts anti-inflammatory effects by regulating TNF-α/Beclin-1 pathway, which provide a new basis for the anti-inflammatory activity of Acorus tatarinowii Schott

lncRNA PRR34-AS1 promotes HCC development via modulating Wnt/β-catenin pathway by absorbing miR-296-5p and upregulating E2F2 and SOX12

Hepatocellular carcinoma (HCC) belongs to the most frequent cancer with a high death rate worldwide. Thousands of long non-coding RNAs (lncRNAs) have been confirmed to influence the development of human cancers, including HCC. Nevertheless, the biological role of PRR34 antisense RNA 1 (PRR34-AS1) in HCC remains obscure. Here, we observed via quantitative real-time reverse transcriptase polymerase chain reaction (quantitative real-time RT-PCR) that PRR34-AS1 was highly expressed in HCC cells. Functional assays revealed that PRR34-AS1 promoted HCC cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) process in vitro and facilitated tumor growth in vivo. In addition, western blot analysis and TOP Flash/FOP Flash reporter assays verified that PRR34-AS1 stimulated Wnt/β-catenin pathway in HCC cells. Furthermore, RNA immunoprecipitation (RIP), RNA pull-down, and luciferase reporter assays uncovered that PRR34-AS1 sequestered microRNA-296-5p (miR-296-5p) to positively modulate E2F transcription factor 2 (E2F2) and SRY-box transcription factor 12 (SOX12) in HCC cells. Importantly, chromatin immunoprecipitation (ChIP) and luciferase reporter assays uncovered that E2F2 transcriptionally activated PRR34-AS1 in turn. Further, rescue experiments reflected that PRR34-AS1 affected HCC progression through targeting miR-296-5p/E2F2/SOX12/Wnt/β-catenin axis. Our findings found that PRR34-AS1 elicited oncogenic functions in HCC, which indicated that PRR34-AS1 might be a novel therapeutic target for HCC