Journal: Journal of Sport and Health Science

Article Title: Exercise training-induced extracellular miR-136-3p modulates glucose uptake and myogenesis through targeting of NRDC in human skeletal muscle

doi: 10.1016/j.jshs.2025.101091

Figure Lengend Snippet: MiR-136-3p content in cell culture medium and uptake of extracellular miR-136-3p into cultured myotubes. MiRNA content in (A) human myotubes and (B) human pancreatic islets culture media. Results were first normalized using RNU1A1 and then presented in relation to miR-23a-3p content for n = 4 different donors for myotubes cultures and n = 4 donors for human islets. Left panel (C) shows bright-field image of cultured human myotubes and right panel (C) shows a representative fluorescence image of cultured human myotubes with cells exposed to human serum-derived EVs loaded with Cy3-miR-136-3p. Cy3 fluorescence (red) is detected in the whole cytoplasm of the human myotubes. (D) Representative fluorescence image of human myotubes exposed to HEK293 culture medium with EVs loaded with Cy3-miR-136-3p (red). (E) Representative image of human myotubes exposed to EVs loaded with TexasRed-labeled with a control RNA (orange). Nuclear Hoechst staining is shown in blue. Scale bar = 100 μm. EVs = extracellular vesicles; HEK293 = human embryonic kidney; miR = microRNA; Rel = relative; RNU1A1 = U1 small nuclear RNA.

Article Snippet: Human embryonic kidney (HEK293) cells were obtained from American Type Culture Collection (ATCC) and cultured in high-glucose (4.5 g/L) Dulbecco's Modified Eagle Medium (DMEM; Thermo Fisher Scientific, Waltham, MA, USA), supplemented with 10% (vol/vol) FBS.

Techniques: Cell Culture, Fluorescence, Derivative Assay, Labeling, Control, Staining

Journal: Journal of Sport and Health Science

Article Title: Exercise training-induced extracellular miR-136-3p modulates glucose uptake and myogenesis through targeting of NRDC in human skeletal muscle

doi: 10.1016/j.jshs.2025.101091

Figure Lengend Snippet: NRDC is a direct target of miR-136-3p in human myotubes. Skeletal muscle NRDC mRNA is responsive to training and inactivity. (A) Tissue mRNA expression of NRDC from the Human Protein Atlas database showing enriched expression of NRDC in human skeletal muscle. (B) The miR-136-3p target site in the NRDC gene is highly conserved in mammals. (C) Luciferase activity in HEK293 cells co-transfected the NRDC 3’UTR and miR-136-3p with or without anti-miR136-3p inhibitors. miR-136-3p transfection downregulates NRDC (D) mRNA and (E) representative image of protein abundance in human myotubes. (F) Publicly available data ( GSE14413 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 6 weeks of endurance training ( n = 8). (G) Publicly available data ( GSE120862 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 2 months of aerobic training ( n = 10). (H) Publicly available data ( GSE14901 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 14 days of immobilization ( n = 24). * p < 0.05, ** p < 0.005. GSE = gene set enrichment; HEK293 = human embryonic kidney; miR = microRNA; NC = negative control; NRDC = nardilysin convertase; nTPM = normalized transcripts per million; si NRDC = small interfering RNA of NRDC ; UTR = untranslated region.

Article Snippet: Human embryonic kidney (HEK293) cells were obtained from American Type Culture Collection (ATCC) and cultured in high-glucose (4.5 g/L) Dulbecco's Modified Eagle Medium (DMEM; Thermo Fisher Scientific, Waltham, MA, USA), supplemented with 10% (vol/vol) FBS.

Techniques: Expressing, Luciferase, Activity Assay, Transfection, Quantitative Proteomics, Negative Control, Small Interfering RNA

Journal: Bioactive Materials

Article Title: Chronic inflammation-responsive hydrogel restores myeloid-T cell crosstalk to reinvigorate antitumor immunity against metastatic colorectal cancer

doi: 10.1016/j.bioactmat.2026.03.012

Figure Lengend Snippet: PGE2 blockade modulates immune cell phenotypes in antitumor resp onses. (A) Inflammatory gene expression across cancer types (GEPIA2 database). (B) Gene expression of Il1b , Cxcl8 , and Lif in colon adenocarcinoma (COAD) tumor tissue and normal tissue (GEPIA2 database). (C and D) Correlation between Ptgs2 and inflammatory genes in various cancers (C) and COAD (D) (TIMER 2.0). (E) Schematic of immune cells co-incubated with CXB treated tumor conditional medium (TCM) (Source material from BioRender). (F and G) Cell viability (F) and Cell cycle arrest (G) detection of CT26 tumor cells treated with gradient concentrations of CXB; n = 3. (H) PGE2 concentration in CT26 cell supernatants; n = 3. (I) The proportion of CD103 + DC within BMDCs after CXB treatments in vitro ; n = 3. (J and K) Maturation (J) and Antigen processing capability (K) on BMDCs; n = 3. (L – N) Flow charts of CD86 or CD206 expression on Raw 264.7 cells (L). Quantification of CD86 (M) and CD206 (N) expression on Raw 264.7 cells; n = 3. (O and P) Flow charts (O) and Quantification (P) of CD69 and CD137 expression on splenic T cells exposed to CXB-pretreated TCM; n = 3. (Q) IFN-γ secretion by T cells co-cultured with CXB-pretreated TCM; n = 3. Data are presented as mean ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Significance was calculated using One-way ANOVA.

Article Snippet: CT26 cells and Raw 264.7 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Gene Expression, Incubation, Concentration Assay, In Vitro, Expressing, Cell Culture

Journal: Bioactive Materials

Article Title: Chronic inflammation-responsive hydrogel restores myeloid-T cell crosstalk to reinvigorate antitumor immunity against metastatic colorectal cancer

doi: 10.1016/j.bioactmat.2026.03.012

Figure Lengend Snippet: Sustained PGE2 blockade prompts immune activ ation. (A) Structure of hydrogel matrix and scheme of Gel-CXB preparation (Source material from BioRender). (B) Microstructure of the hydrogel. (C) Rheological evaluation of Gel-CXB. (D) CXB release from Gel-CXB in PBS or PBS containing 0.5 mM H 2 O 2 ; n = 3. (E and F) Flow chart (E) and Quantification (F) of CD103 + DC within BMDCs; n = 3. (G and H) Flow chart (G) and Heatmap (H) of costimulatory molecular expression on CD103 - DC, CD103 + DC, or total DC with different treatments; n = 3. (I and J) CXCL9 (I) and Costimulatory molecular expression (J) on cDC1; n = 3. (K – M) CD86 and CD206 expression (K), MHC-II expression (L), and Antigen processing capability (M) of BMDMs incubated with different TCM; n = 3. (N and O) CD69 (N) and CD137 (O) expression on CD8 + T cells co-incubated with different TCM; n = 3. (P) Scheme of Gel-CXB-regulated CT26 TME at different time points in vivo . (Q) Changes of several immune cells within TME at Day 1, 5, and 9; n = 3. (R) Tumor volume of mice treated with CXB alone or Gel-CXB in vivo ; n = 5. (S) CD137 expression on CD8 + T cells in vivo ; n = 3. Data are presented as mean ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Significance was calculated using One-way ANOVA.

Article Snippet: CT26 cells and Raw 264.7 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Expressing, Incubation, In Vivo

Journal: Bioactive Materials

Article Title: Chronic inflammation-responsive hydrogel restores myeloid-T cell crosstalk to reinvigorate antitumor immunity against metastatic colorectal cancer

doi: 10.1016/j.bioactmat.2026.03.012

Figure Lengend Snippet: TRANS inhibits tumor growth and enhances local and systemic immune resp onses. (A) Scheme of GC, GCF, or TRANS preparation (Source material from BioRender). (B) Microstructure of GC and TRANS. (C) Experimental design for administration and immune cell analysis. (D) Tumor growth curves under different treatments; n = 5. (E) Tumor weight post-treatment; n = 5. (F – H) CD45 + leukocytes and CD11c + DCs (F), CD86 + M1 and CD206 + M2 macrophages (G), and Tumor-infiltrating CD8 + T cells (H) within TME; n = 5. (I – L) Mature DCs (I), CD8α + cDC1s (J), CD4 + and CD8 + T cells (K) and CD69 + CD8 + T cells (L) in lymph nodes; n = 5. (M – Q) CD11c + MHC II + DCs (M), CD8α + cDC1s (N), CD4 + and CD8 + T cells (O), CD69 + CD8 + T cells (P), and IFN-γ + CD8 + T cells (Q) in the spleen; n = 5. (R – T) CD8 + T cells (R), The ratio of CD8 + T /CD4 + T cells (S), and IFN-γ levels (T) in blood; n = 5. (U) IFN-γ + CD4 + T and IFN-γ + CD8 + T cells with ex vivo stimulation of PMA/ionomycin for 6 h; n = 3. (V) Apoptosis of CT26 cells co-incubated with splenic T cells for 24 h; n = 3. Data are presented as mean ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Significance was calculated using One-way ANOVA.

Article Snippet: CT26 cells and Raw 264.7 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Cell Analysis, Ex Vivo, Incubation

Journal: Bioactive Materials

Article Title: Chronic inflammation-responsive hydrogel restores myeloid-T cell crosstalk to reinvigorate antitumor immunity against metastatic colorectal cancer

doi: 10.1016/j.bioactmat.2026.03.012

Figure Lengend Snippet: TRANS inhibits tumor metastasis and induces immune memory in vivo . (A) Experimental design for secondary tumor model. (B and C) Tumor volume curves of primary tumor (B) and secondary tumor (C) during different therapy; n = 5. (D and E) Statistical diagram (D) and flow charts (E) of T cells within secondary tumors; n = 5. (F) Immunofluorescence images of immune cell in primary tumor. (G) Schematic of lung metastasis tumor model and treatment regimen. Mice received subcutaneous and intravenous injections of CT26-Luc. (H – J) In vivo images (H), Primary tumor volume curves (I), and Average radiance in lungs (J) of CT26-Luc tumor-bearing mice; n = 5. (K – M) Lung image (K), Lung metastasis foci counts and weights (L), and H&E staining of lungs (M) from CT26-Luc tumor-bearing mice; n = 5. (N) Schematic of liver metastasis tumor model and treatment regimen. Mice received subcutaneous CT26 tumor and splenic CT26-Luc injections. (O and P) In vivo imaging (O) and Individual radiance in livers (P) of CT26-Luc tumor-bearing mice; n = 10. (Q – S) Live images (Q), Liver weights (R), and H&E staining images of livers (S) from PBS- or TRANS-treated mice; n = 5. (T) Scheme of tumor rechallenge model. (U) Tumor changes in mice rechallenged with CT26 or 4T1; n = 9. (V) Central memory (T CM ) and effector memory (T EM ) gated on CD8 + T cells; n = 5. Data are presented as mean ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Significance was calculated using One-way ANOVA.

Article Snippet: CT26 cells and Raw 264.7 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: In Vivo, Immunofluorescence, Staining, In Vivo Imaging