Objective-The compositions of the gut microbiota and its metabolites were altered in individuals with Autism Spectrum Disorder (ASD). The aim of this study was to assess whether plasma levels of gut-derived metabolite trimethylamine N-oxide (TMAO) were associated with ASD and the degree of symptom severity. Methods-From September 2017 to January 2019, a total of three hundred and twenty-eight Chinese children (164 with ASD and 164 their age-sex matched control subjects) aged 3-8 years were included. TMAO levels in plasma were determined using high-performance liquid chromatography tandem mass spectrometry (LC/MS/ MS). Logistic regression analysis was used to examine the TMAO-ASD association.

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IntroductionMiR-101 rs7536540 may influence the expression of miR-101 and another polymorphism, rs2227513, which in turn affects the expression of IL-22, . However, studies on the combined effect of these polymorphisms are still scarce.Material and methodsQuantitative real-time PCR was performed to analyze the expression of miR-101 and IL-22 mRNA. ELISA and Western blot were carried out to examine the expression of IL-22 protein. MTT assay and flow cytometry were used to assess the cellular proliferation and apoptosis . Immunofluorescence was performed to measure the expression of p-STAT3. Luciferase assay was carried out to explore the inhibitory role of miR-101 in the expression of IL-22.ResultsThe severity of RA was progressively increased in patients with rs7536540 GG + rs2227513 AA, rs7536540 GG + rs2227513 AG, rs7536540 CC/CG + rs2227513 AA and rs7536540 CC/CG + rs2227513 AG genotypes. The CC/CG alleles at rs7536540 were correlated with up-regulated miR-101 expression in the serum and SF of RA patients, whereas both CC/CG alleles at rs7536540 and AG alleles at rs2227513 were correlated with elevated expression of IL-22. Incubation of FLS with SF isolated from RA patients carrying the CC/CG alleles at rs7536540 and AG alleles at rs2227513 remarkably increased the cell proliferation and inhibited the apoptosis of FLS. Luciferase assay demonstrated that the expression of IL-22 was notably suppressed by miR-101 in THP-1 cells.ConclusionsOur study revealed the combined effect of polymorphisms rs7536540 and rs2227513 on the expression of IL-22 and the proliferation of FLS as well as their association with the severity of RA