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Butyric acid (BA) can potentially enhance the function of the intestinal barrier. However, the mechanisms by which BA protects the intestinal mucosal barrier remain to be elucidated. Given that the Ras homolog gene family, member A (RhoA)/Rho-associated kinase 2 (ROCK2)/Myosin light chain kinase (MLCK) signaling pathway is crucial for maintaining the permeability of the intestinal epithelium, we further investigated whether BA exerts a protective effect on epithelial barrier function by inhibiting this pathway in LPS-induced Caco2 cells. First, we aimed to identify the optimal treatment time and concentration for BA and Lipopolysaccharide (LPS) through a CCK-8 assay. We subsequently measured Trans-epithelial electrical resistance (TEER), FITC-Dextran 4 kDa (FD-4) flux, and the mRNA expression of ZO-1, Occludin, RhoA, ROCK2, and MLCK, along their protein expression levels, and average fluorescence intensity following immunofluorescence staining. We then applied the ROCK2 inhibitor Y-27632 and reevaluated the TEER, FD-4 flux, and mRNA, and protein expression of ZO-1, Occludin, RhoA, ROCK2, and MLCK, as well as their distribution in Caco2 cells. The optimal treatment conditions were determined to be 0.2 mmol/L BA and 5 μg/mL LPS for 24 hours. Compared with LPS treatment alone, BA significantly mitigated the reduction in the TEER, decreased FD-4 flux permeability, increased the mRNA expression of ZO-1 and Occludin, and normalized the distribution of ZO-1 and Occludin in Caco2 cells. Furthermore, BA inhibited the expression of RhoA, ROCK2, and MLCK, and normalized their localization within Caco2 cells. Following treatment with Y-27632, the epithelial barrier function, along with the mRNA and protein expression and distribution of ZO-1 and Occludin were further normalized upon inhibition of the pathway. These findings contribute to a deeper understanding of the potential mechanisms through which BA attenuates LPS-induced impairment of the intestinal epithelial barrier. [ABSTRACT FROM AUTHOR] more...

Rhizoctonia solani AG-1 IA is one of the most important fungal pathogens that causes severe yield losses in rice-growing regions worldwide. Catechol (1, 2-benzenediol) is a natural catecholamine which can induce melanization of fungi and contribute to enhanced virulence and fungal resistance to diverse environments. In this study, the effects of catechol on growth, activities of antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), glutathione S-transferase (GSH-ST), glutathione peroxidase (GSH-PX) and glutathione reductase (GR), as well as expression of melanin biosynthesis genes in R. solani AG-1 IA, were evaluated by culturing the fungus in catechol-containing nutrient media. The results showed that the activities of CAT, POD, SOD, GSH-ST, GSH-PX and GR were decreased in the presence of 12.5-50 μg mL−1 catechol, while GSH-PX activity was significantly increased (by 310%) in the presence of 50 µg mL−1 catechol. The growth rates of R. solani AG-1 IA on potato dextrose agar (PDA) containing catechol were significantly decreased, whereas the pathogenicity to rice of R. solani AG-1 IA cultured on PDA containing different concentrations of catechol was enhanced when compared with the catechol-free control. Quantitative RT-PCR results showed that the expression of four melanin biosynthesis genes, i.e. protein tyrosine phosphatase (Ptp), chorismate mutase (Cm), peroxidase (Per) and phenol hydroxylase (Ph), was significantly down-regulated in R. solani AG-1 IA cultured in catechol-containing PDB at 100 µg mL−1. These results suggest that catechol was utilized by the fungus to synthesize melanin, and it could also induce the activities of antioxidant enzymes and melanization in R. solani AG-1 IA. Our results provide strong evidence to support the view that melanin contributes to virulence directly through fungal antioxidant defence systems in which melanization is induced by oxidative stress. [ABSTRACT FROM AUTHOR] more...

Objective: To investigate the HLA-DRB1, DQB1 allele polymorphism in Kunming Yi nationality population., Methods: HLA-DRB1, DQB1 DNA types in 70 healthy children of Yi nationality in Kunming were analyzed by polymerase chain reaction with sequence specific primer (PCR-SSP)., Results: Twelve alleles at HLA-DRB1 locus were observed in the 70 children: the alleles with gene frequencies higher than 10% were HLA-DRB1*12(33.57%), DRB1*0901(11.43%), DRB1*04(11.43%); the alleles with gene frequencies between 10% and 5% were HLA-DRB1*01(8.57%), DRB1*11(7.86%), DRB1*14(7.14%), DRB1*15(7.14%), DRB1*08(5%); the alleles with gene frequencies lower than 5% were HLA-DRB1*03(2.86%), DRB1*13(2.14%), DRB1*07(1.43%), DRB1*16(1.43%). Seven alleles at HLA-DQB1 locus were observed in the 70 children: the alleles with gene frequencies higher than 10% were HLA-DQB1*0301(45%), DQB1*05(22.14%), DQB1*0303(12.14%); the alleles with gene frequencies between 10% and 5% were HLA-DQB1*04(6.43%), DQB1*06(6.43%); the alleles with gene frequencies lower than 5% were HLA-DQB1*0201(4.29%) and DQB1*0302(3.57%)., Conclusion: The distribution of HLA-DRB1, DQB1 allele polymorphism in the Kunming Yi nationality population is distinctive. It is neither like that in the South Han population nor like that in the North Han population. more...