Regulatory effects of mangiferin on LPS-induced inflammatory responses and intestinal flora imbalance during sepsis.

Studies suggest that mangiferin (MAF) has good therapeutic effects on chronic bronchitis and hepatitis. Also, it is one of the antiviral ingredients in Anemarrhena asphodeloides Bunge. However, its effect on the LPS-induced inflammation and intestinal flora during sepsis remains unclear yet. In the present study, LPS-stimulated inflammation RAW264.7 cells and LPS-induced sepsis mice were used to evaluate the efficacy of MAF in vitro and in vivo. 16S rDNA sequencing was performed to analyze the characteristics of intestinal flora of the sepsis mice. It has been demonstrated that MAF (12.5 and 25 µg/mL) significantly inhibited protein expressions of TLR4, MyD88, NF-κB, and TNF-α in the LPS-treated cells and reduced the supernatant TNF-α and IL-6 levels. In vivo, MAF (20 mg/kg) markedly protected the sepsis mice and reduced the serum TNF-α and IL-6 levels. Also, MAF significantly downregulated the protein expressions of TLR4, NF-κB, and MyD88 in the livers. Importantly, MAF significantly attenuated the pathological injuries of the livers and small intestines. Further, MAF significantly increased proportion of Bacteroidota and decreased the proportions of Firmicutes, Desulfobacterota, Actinobacteriota, and Proteobacteria at phylum level, and it markedly reduced the proportions of Escherichia-Shigella, Pseudoalteromonas, Staphylococcus at genus level. Moreover, MAF affects some metabolism-related pathways such as citrate cycle (TCA cycle), lipoic acid metabolism, oxidative phosphorylation, bacterial chemotaxis, fatty acid biosynthesis, and peptidoglycan biosynthesis of the intestinal flora. Thus, it can be concluded that MAF as a treatment reduces the inflammatory responses in vitro and in vivo by inhibiting the TLR4/ MyD88/NF-κB pathway, and corrects intestinal flora imbalance during sepsis to some degree.

Potent salinomycin C20-O-alkyl oxime derivative SAL-98 efficiently inhibits tumor growth and metastasis by affecting Wnt/ß-catenin signal pathway.

The dysregulation of Wnt/ß-catenin signaling pathway is closely related to tumorigenesis, metastasis and cancer stem cell maintenance. Salinomycin is a polyether ionophore antibiotic that selectively eliminates cancer stem cells by inhibiting the Wnt/ß-catenin signal pathway. Salinomycin selectively target cancer stem cells, but the toxicity limits its further use. In this study, we explore the anti-tumor mechanism of one most active salinomycin C20-O-alkyl oximederivative SAL-98 and found that SAL-98 exerts 10 times higher anti-tumor and anti-CSCs activities compared with salinomycin, which induces cell cycle arrest, ER stress and mitochondria dysfunction and inhibits Wnt/ß-catenin signal pathway in vitro with high efficacy. Moreover, SAL-98 shows good anti-metastasis effect in vivo. In addition, SAL-98 demonstrates same anti-tumor activities as salinomycin with less 5 times concentration in vivo, the ER stress, autophagy and anti-CSCs effects were also confirmed in vivo. Mechanistically, SAL-98 inhibits the Wnt/ß-catenin signaling pathway associated with CHOP expression induced by ER stress, the induced CHOP disrupts the ß-catenin/TCF4 complex and represses the Wnt targeted genes. This study provides an alternative strategy for rational drug development to target Wnt/ß-catenin signaling pathway.

Clinical application, evaluation and analysis of influencing factors of on tuberculosis γ-Interferon enzyme-linked immunosorbent assay.

OBJECTIVE: To assess sensitivity and specificity of the interferon gamma release assay test, and to pinpoint the influencing factors that should be taken care of in clinical application. METHODS: This study was conducted at the First People's Hospital in the Yunnan Province of China from October 2018 to March 2019, and comprised samples collected from outpatient and inpatients. To detect mycobacterium tuberculosis, acid-fast staining on sputum smear was performed on relevant tissues suspected of extrapulmonary tuberculosis. Tuberculosis interferon gamma release assay test and pathology samples were examined. Tuberculosis-specific cell reaction assay kit was used for sampling. SysmexXN-2000 haematology analyser, VACUETTE SRS100/II and Beckman Coulter AU5800 were used to perform various analyses. Data was grouped and analysed using R statistical software. RESULTS: Of the 960 samples, 516(53.75%) cases tested positive for tuberculosis infection and 444(46.25%) tested negative. The sensitivity of the pathological results was 86% and the specificity was 96%. The sensitivity of the interferon gamma release assay test was 95% and specificity 82%. Interferon release test, pathological results and final diagnosis showed significant comparisons (p<0.05). Significant relationships were also established for factors, such as age, interferon release quantity, lymphocyte, C-reactive protein and counts of mono-nuclear cell (p<0.05). CONCLUSION: Interferon gamma release assay test was found to have high consistency with pathological results and final diagnosis and can be used as a subsidiary to traditional clinical imaging and pathological judgment.