Fig 1: Safety and toxicity evaluation of the active immunotherapy At the top, representative images of H&E staining of two vaccinated and two sham mice show normal-looking structure of kidneys from sham and vaccinated mice, as indicated. Arrows show glomeruli and arrowheads show tubuli. At the bottom, cleaved-Caspase3 staining on kidney sections from sham and vaccinated mice is shown, with no sign of damage (each biological replicate was run in five technical replicates). Apoptotic cells are indicated by arrows. Scale bar = 50 µm.Serum CRP concentrations were performed in duplicate in sham (n = 9) and vaccinated (n = 8) mice as determined by ELISA. Statistical significance was assessed by unpaired Student's t-test with welch's correction (Vaccinated vs. Sham ns P = 0.7002, ns = non-significant). Error bars indicate standard error of the mean (SEM).Serum Neurofilament Light chain concentrations in sham (n = 8), vaccinated (n = 8), and non-vaccinated (n = 12) mice was determined by SIMOA. Statistical significance was assessed using an ordinary one-way ANOVA followed by Tukey's multiple comparisons test (Sham vs. Vaccinated ns P = 0.9974; Sham vs. 7 m.o. ns P = 0.518; Vaccinated vs. 7 m.o. ns P = 0.5638, ns = non-significant). Error bars indicate standard error of the mean (SEM). Source data are available online for this figure.
Fig 2: Prostant regulated inflammatory factors expression through modulating Th17/Treg balance with inhibiting JAK/STAT axis. (a) The serum level of hs-CRP, TNF-a, IL-2, and IL-6 was examined via ELISA. ((b) and (c)) The level of VEGF in mucosal tissues was determined by IHC. ((d) and (e)) The expression of Th17 and Treg was assessed by flow cytometry assay. ((f) and (g)) The protein expression of STAT3, phosphorylated STAT3, SOCS3, and JAK2 was evaluated by western blot. Data were exhibited following being normalized to GAPDH. The means ± SD of six independent samples were displayed. **p < 0.01 and ***p < 0.001 v.s control group. #p < 0.05, ##p < 0.01 and ###p < 0.001 v.s H&UR model group.
Fig 3: Effect of dietary treatment and influence of exercise upon inflammatory biomarkers in liver and muscle in C57BL/6J mice. Liver concentration of (A) C-Reactive Protein (CRP, pg/mL); (B) Vascular Endothelial Growth Factor (VEGF, pg/mL); (C) Interleukin 6 (IL-6, pg/mL): (D) NF kappa ß p65, (pS536 + Total), pg/mL; (E) Caspase-3 (OD@450 nm); and, muscle concentration of (F) Tumor Necrosis Factor, (TNF-a, pg/mL). Mice were provided these 5 experimental diets: (1) 0 mg/kg folic acid FA/17.9% protein, TD.150200, FA Deficient (D) Diet (FAD; n = 16); (2) 2 mg/kg FA/17.9% protein; TD.150201, Control (C) FA Protein (P) Diet (FPC; n = 12); (3) 7 mg/kg, FA/17.9% protein; TD.15202, FA Supplemented (S) Diet (FAS, n = 16); (4) 2 mg/kg FA, 6% protein [Low Protein Diet (LPD, n = 16)]; and (5) 2 mg/kg FA/48% protein [High Protein Diet (HPD, n = 16)]. The results are means ? SEM (n = 8). Panel (B) VEGF p < 0.05, exercised FPC vs. HPD; panel (D) NF kappa ß p65 p < 0.05, exercised FPC vs. HPD; panel (E) Caspase-3 p < 0.05, exercised FPC vs. HPD; and, panel (F) muscle TNF-a, unexercised FPC and FAS, p < 0.05.
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