Additional file 1 of Effect of arsenic on the risk of gestational diabetes mellitus: a systematic review and meta-analysis

Supplementary Material 1: Search strategy for electronic databases

Fine root dynamics data sheat (Jamro et al. 2015)

Comma-separated values (CSV

Inhibition of dsDNA-Templated Copper Nanoparticles by Pyrophosphate as a Label-Free Fluorescent Strategy for Alkaline Phosphatase Assay

On the basis of the inhibition of double strand DNA (dsDNA)-templated fluorescent copper nanoparticles (CuNPs) by pyrophosphate (PPi), a novel label-free turn-on fluorescent strategy to detect alkaline phosphatase (ALP) under physiological conditions has been developed. This method relies on the strong interaction between PPi and Cu²⁺, which would hamper the effective formation of fluorescent CuNPs, leading to low fluorescence intensity. The ALP-catalyzed PPi hydrolysis would disable the complexation between Cu²⁺ and PPi, facilitating the formation of fluorescent CuNPs through the reduction by ascorbate in the presence of dsDNA templates. Thus, the fluorescence intensity was recovered, and the fluorescence enhancement was related to the concentration of ALP. This method is cost-effective and convenient without any labels or complicated operations. The present strategy exhibits a high sensitivity and the turn-on mode provides a high selectivity for the ALP assay. Additionally, the inhibition effect of phosphate on the ALP activity was also studied. The proposed method using a PPi substrate may hold a potential application in diagnosis of ALP-related diseases or evaluation of ALP functions in biological systems

Differentially expressed genes analysis, GO and KEGG pathway enrichment and enrichment and among groups.

(A–F): DEGs of rats; A and B: model vs. control; C and D: medium-dose SYNC vs. model; E and F: PI vs. model. (G): Venn diagrams. Expression of Nr4a3 (H) and PI3K (I) in individual samples and STRING protein interaction network map(J). SYNC therapeutic genes are associated with biological functions (K), enrichment bubble map (L) and differential gene KEGG pathway enrichment analysis bubble map (M).</p

Toluidine blue and immunohistochemical staining results of rat colon tissue (400x).

MCs are blue-purple after toluidine blue staining. SYNC reduced MCs infiltration and inhibited colonic MCT expression. (A-B) Number of MCs in the colon (6 randomly selected 400x magnification fields). (C-D) MCT expression in the colon. (* p p 0.01 vs. model group; # p p < 0.01 vs. control group).</p

S1 Raw images -

Aim of the studyTo evaluate the therapeutic effect of SYNC in diarrhea irritable bowel syndrome (IBS-D) and explore its underlying mechanism through transcriptomic sequencing (RNA-Seq).Materials and methodsA rat model of IBS-D was constructed to elucidate the effects of SYNC. Abdominal withdrawal reflex (AWR), fecal water content (FWC), and recording body weight were calculated to assess visceral sensitivity in rats. Histopathological changes in the colon and alterations in mast cell (MC) count were determined. Immunohistochemistry was employed to assess mast cell tryptase (MCT) expression in rat colons. Serum levels of corticotropin-releasing Hormone (CRH), interleukin-6 (IL-6), calcitonin gene-related peptide (CGRP), and 5-hydroxytryptamine (5-HT) were quantified using ELISA. RNA-Seq of colon tissue was performed, followed by Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Western blot analysis was conducted to quantify the expression levels of key proteins in the Nr4a3 pathway in the colon and hypothalamus tissues of rats.ResultsSYNC alleviated visceral hypersensitivity and mood disorders in rats with IBS-D. Moreover, it was positively correlated with its dosage and the observed effects, such as the enhancement of the colon’s mucosal lining condition and reduction in the number and activation of MCs within the model group. SYNC reduced the expression levels of factors related to the brain-gut axis and inflammatory markers in the bloodstream. RNA-Seq analysis indicated that SYNC down-regulated the expression of Nr4a3 and PI3K. These SYNC-targeted genes primarily played roles in immune regulation and inflammatory responses, correlating with the modulation of Nr4a3 and the PI3K/AKT pathway. Western blot analysis further confirmed SYNC’s influence on inflammation-related MC activation by downregulating key proteins in the Nr4a3/PI3K pathway.ConclusionsSYNC inhibited mast cell activation and attenuated visceral hypersensitivity in the colon tissues of IBS-D rats. These effects were mediated by the Nr4a3/PI3K signaling pathway.</div

S1 Data -

Aim of the studyTo evaluate the therapeutic effect of SYNC in diarrhea irritable bowel syndrome (IBS-D) and explore its underlying mechanism through transcriptomic sequencing (RNA-Seq).Materials and methodsA rat model of IBS-D was constructed to elucidate the effects of SYNC. Abdominal withdrawal reflex (AWR), fecal water content (FWC), and recording body weight were calculated to assess visceral sensitivity in rats. Histopathological changes in the colon and alterations in mast cell (MC) count were determined. Immunohistochemistry was employed to assess mast cell tryptase (MCT) expression in rat colons. Serum levels of corticotropin-releasing Hormone (CRH), interleukin-6 (IL-6), calcitonin gene-related peptide (CGRP), and 5-hydroxytryptamine (5-HT) were quantified using ELISA. RNA-Seq of colon tissue was performed, followed by Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Western blot analysis was conducted to quantify the expression levels of key proteins in the Nr4a3 pathway in the colon and hypothalamus tissues of rats.ResultsSYNC alleviated visceral hypersensitivity and mood disorders in rats with IBS-D. Moreover, it was positively correlated with its dosage and the observed effects, such as the enhancement of the colon’s mucosal lining condition and reduction in the number and activation of MCs within the model group. SYNC reduced the expression levels of factors related to the brain-gut axis and inflammatory markers in the bloodstream. RNA-Seq analysis indicated that SYNC down-regulated the expression of Nr4a3 and PI3K. These SYNC-targeted genes primarily played roles in immune regulation and inflammatory responses, correlating with the modulation of Nr4a3 and the PI3K/AKT pathway. Western blot analysis further confirmed SYNC’s influence on inflammation-related MC activation by downregulating key proteins in the Nr4a3/PI3K pathway.ConclusionsSYNC inhibited mast cell activation and attenuated visceral hypersensitivity in the colon tissues of IBS-D rats. These effects were mediated by the Nr4a3/PI3K signaling pathway.</div

A flowchart of the model development protocol.

Aim of the studyTo evaluate the therapeutic effect of SYNC in diarrhea irritable bowel syndrome (IBS-D) and explore its underlying mechanism through transcriptomic sequencing (RNA-Seq).Materials and methodsA rat model of IBS-D was constructed to elucidate the effects of SYNC. Abdominal withdrawal reflex (AWR), fecal water content (FWC), and recording body weight were calculated to assess visceral sensitivity in rats. Histopathological changes in the colon and alterations in mast cell (MC) count were determined. Immunohistochemistry was employed to assess mast cell tryptase (MCT) expression in rat colons. Serum levels of corticotropin-releasing Hormone (CRH), interleukin-6 (IL-6), calcitonin gene-related peptide (CGRP), and 5-hydroxytryptamine (5-HT) were quantified using ELISA. RNA-Seq of colon tissue was performed, followed by Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Western blot analysis was conducted to quantify the expression levels of key proteins in the Nr4a3 pathway in the colon and hypothalamus tissues of rats.ResultsSYNC alleviated visceral hypersensitivity and mood disorders in rats with IBS-D. Moreover, it was positively correlated with its dosage and the observed effects, such as the enhancement of the colon’s mucosal lining condition and reduction in the number and activation of MCs within the model group. SYNC reduced the expression levels of factors related to the brain-gut axis and inflammatory markers in the bloodstream. RNA-Seq analysis indicated that SYNC down-regulated the expression of Nr4a3 and PI3K. These SYNC-targeted genes primarily played roles in immune regulation and inflammatory responses, correlating with the modulation of Nr4a3 and the PI3K/AKT pathway. Western blot analysis further confirmed SYNC’s influence on inflammation-related MC activation by downregulating key proteins in the Nr4a3/PI3K pathway.ConclusionsSYNC inhibited mast cell activation and attenuated visceral hypersensitivity in the colon tissues of IBS-D rats. These effects were mediated by the Nr4a3/PI3K signaling pathway.</div

SYNC inhibits colonic Nr4a3, p-PI3K, p-AKT and CRH-R1 expression, and hypothalamic CRH-R1 expression in IBS-D rats.

Expression of key proteins in the rat (A). Western blot quantification of PI3K (B), AKT (C), Nr4a3 (D), p-PI3K (E), p-AKT (F) colon CRH-R1 (H) and hypothalamus CRH-R1 (J) in the colon and hypothalamus of rats for different groups. (* p p 0.01 vs. model group; # p p<0.01 vs. control group, n = 3).</p

SYNC reduces anxiety and depression in IBS-D rats.

The times of entries into the central (A) and surrounding zones (B) as well as the times of grooming (C) and stand-up (D) before and after treatment were recorded. The sugar-water bias rate of the SYNC group was significantly different from that of the model group (E). One-way ANOVA was used for statistical analysis between different groups. (* p p 0.01 vs. model group; # p 0.05, # # p 0.01 vs. control group, n = 8).</p