Sarcopenia is a predictive factor for intestinal resection in admitted patients with Crohn’s disease - Fig 2

<p>Cumulative operation-free survival rate for all patients (a), patients with Crohn’s disease (b), patients with ulcerative colitis (c).</p

Factors associated with intestinal resection for all IBD patients.

<p>Factors associated with intestinal resection for all IBD patients.</p

PCR primers used in this study.

<p>PCR primers used in this study.</p

Changes of SMI after intestinal resection.

<p>Solid and dashed line indicates patients with CD and UC, respectively. Double vertical lines indicate intestinal resection. AZA: azathioprine.</p

mRNA expression of cytokines in the colon.

<p>Real-time PCR analysis for the mRNA expression of cytokines was performed on the colonic mucosa. The cytokine mRNA expression was converted to a value relative to β-actin mRNA expression, and was presented as an increase relative to the results for control mice (no treatment). Data are expressed as means ± SD of five different samples. *<i>P</i> < 0.05, **<i>P</i> < 0.01.</p

Effects of IL-1β on IL-36γ mRNA expression in colonic myofibroblasts.

<p>(A) Dose-dependent effects of IL-1β on IL-36γ mRNA expression. The cells were incubated for 24 h with increasing concentrations of IL-1β. IL-36γ mRNA expression was expressed relative to β-actin mRNA expression (mean ± SD from 4 different experiments). (B) Time-dependent effects of IL-1β on IL-36γ mRNA expression. The cells were stimulated with IL-1β (10 ng/ml) for the pre-determined times. IL-36γ mRNA expression was expressed relative to β-actin mRNA expression (mean ± SD from 4 different experiments). (C) Dose-dependent effects of IL-1β on IL-36γ secretion. The cells were incubated for 24 h with increasing concentrations of IL-1β. IL-36γ level in supernatant was determined by ELISA (mean ± SD from 4 different experiments). (D) Time-dependent effects of IL-1β on IL-36γ secretion. The cells were stimulated with IL-1β (10 ng/ml) for the pre-determined times. IL-36γ level was determined by ELISA (mean ± SD from 4 different experiments). *P<0.05, **P<0.01 versus medium; ANOVA followed by Bonferroni’s post hoc test.</p

The expression of cytokines in the colon tissue.

<p>The expression of IL-1β, IL-6, IL-17A, IFN-γ, and IL-10 in the colon tissues was examined using immunohistochemistry. Control staining was also presented. The data are representative of four independent experiments. Magnification x 100.</p

Proportion of Treg cells in the lamina propria of the colon.

<p>(A) Flow cytometric analysis for Foxp3⁺CD4⁺ T reg cells in the colonic lamina propria. Data are representative of five independent experiments. (B) Proportion of Foxp3⁺CD4⁺ T reg cells. Data are expressed as means ± SD of five different samples. *<i>P</i> < 0.05, **<i>P</i> < 0.01, n.s.; not significant.</p

Effect of TN-3 on the concentration of fecal SCFAs.

<p>The concentrations of fecal SCFAs were measured by High-performance liquid chromatography: (A) the concentration of butyrate, (B) the concentration of acetate, (C) the concentration of propionate. Data are expressed as means ± SD of five different samples. *<i>P</i> < 0.05, **<i>P</i> < 0.01, n.s.; not significant.</p

Combined effects of IL-1β and other cytokines.

<p>(A) The cells were incubated for 24 h with combination of IL-1β (10 ng/ml) and other cytokines [TNF-α (100 ng/ml), IFN-γ (100 ng/ml), IL-4 (100 ng/ml), and/or IL-17A (100 ng/ml)]. IL-36γ mRNA expression was analyzed by real-time PCR. IL-36γ mRNA expression was expressed relative to the β-actin mRNA expression (mean ± SD from 4 different experiments). **P<0.01 versus IL-1β alone. (B) The cells were stimulated for 24 h with or without IL-1β (10 ng/ml), TNF-α (100 ng/ml), and/or combination of IL-1β (10 ng/ml) and TNF-α (100 ng/ml), and intracellular IL-36γ was analyzed by Western blot.</p