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: MiR-136-3p was labeled with Cy3 using Silencer small interfering RNA (siRNA) Labeling Kit with Cy3 Dye (Thermo Fisher Scientific) and loaded into exosome-enriched EVs with Exo-Fect siRNA/miRNA Transfection Reagent (System Biosciences, Palo Alto, CA, USA).

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

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: Cellular metabolism in human myotubes after miR-136-3p transfection or NRDC silencing. Mitochondrial respiration in miR-136-3p-transfected or NRDC- silenced human myotubes was monitored using the Mitochondrial Stress Test. (A) OCR and (B) ECAR were measured using the Seahorse XFe24 Extracellular Flux Analyzer. The trace shows representative data. (C) Quantification of the mitochondrial respiration data for basal respiration, maximal respiration, ATP production, and spare respiratory capacity obtained from 3 independent experiments. Human myotubes were transfected with miR-136-3p or siRNA against NRDC before determination of (D) uptake of radiolabeled glucose, (E) rates of radiolabeled glucose oxidation, (F) conversion of radiolabeled glucose into glycogen, (G) rate of radiolabeled palmitic acid oxidation, (H) protein synthesis as assessed by incorporation of puromycin, and (I) lactate release into the media. Results are expressed as mean ± standard error of the mean. * p < 0.05, ** p < 0.005 vs. control cells. ECAR = extracellular acidification rate; FCCP = carbonyl cyanide-p-trifluoromethoxyphenylhydrazone; miR = microRNA; NC = negative control; NRDC = nardilysin convertase; ns = no significance; OCR = oxygen consumption rate; OigoA = oligomycin A; Rot/AA = rotenone and antimycin A; si NRDC = small interfering RNA of NRDC; siRNA = small interfering RNA; scr = negative control for small interfering RNA.

Article Snippet: MiR-136-3p was labeled with Cy3 using Silencer small interfering RNA (siRNA) Labeling Kit with Cy3 Dye (Thermo Fisher Scientific) and loaded into exosome-enriched EVs with Exo-Fect siRNA/miRNA Transfection Reagent (System Biosciences, Palo Alto, CA, USA).

Techniques: Transfection, Control, Negative Control, Small Interfering RNA

Journal: Journal of Sport and Health Science

Article Title: Long-term aerobic exercise enhances circulating exosomal miR-214-3p to promote endothelial progenitor cell-mediated repair of endothelial damage induced by obesity

doi: 10.1016/j.jshs.2025.101094

Figure Lengend Snippet: Circulating exosomal miR-214-3p regulated the function of EPC in both humans and rats with obesity. (A and B) Targeted transcriptome sequencing of miRNAs within circulating exosomes derived from (A) humans and (B) rats unveiled the regulatory impact of aerobic exercise on miRNA expression profiles in these vesicles. The left panel shows a volcano plot of differentially expressed genes, and the right panel shows a heatmap representation of the transcriptome sequencing data, illustrating the expression patterns of differentially expressed genes in circulating exosomes. (C and D) qPCR was used to validate the relative expression levels of miR-214-3p in circulating exosomes derived from (C) humans ( n = 3 for each group; * p < 0.05, Exercise vs . Control) and (D) rats ( n : 6–12 for each group; * p < 0.05, HC vs . NC; # p < 0.05, HE vs . HC). (E) qPCR validation of miR-214-3p expression in miR-214-3p-overexpressing EPC ( n = 3 for each group). ** p < 0.01, miR-214-3p mimics vs . mimics NC; ## p < 0.01, miR-214-3p mimics vs . Control. (F) Summary data for cell vitality level among groups ( n : 10–12 for each group). *** p < 0.001, miR-214-3p mimics vs . mimics NC. (G and H) Summary data (G) and representative images (H) of wound healing in the scratch assay, showcasing the migration level of EPC among groups ( n = 4 for each group). * p < 0.05, miR-214-3p mimics vs . mimics NC. EPC = endothelial progenitor cells; HC = the high-fat diet with sedentary group; HE = the high-fat diet with exercise group; mimics NC = mimics negative control; miR = microRNA; NC = the normal diet with sedentary group; qPCR = quantitative polymerase chain reaction.

Article Snippet: Total RNAs from tissues, cells, and exosomes were extracted using Trizol (R0016; Beyotime Biotech, Shanghai, China), following the manufacturer’s recommendations. mRNA samples underwent reverse transcription using the Evo M-MLV Kit (AG11705; Accurate Biotechnology, Changsha, China), while miRNA samples were reversely transcribed using the All-in-OneTM miRNA qPCR Kit (QP115; iGene Biotechnology, Guangzhou, China), following the provided instructions.

Techniques: Sequencing, Derivative Assay, Expressing, Control, Biomarker Discovery, Wound Healing Assay, Migration, Negative Control, Real-time Polymerase Chain Reaction

Journal: Journal of Sport and Health Science

Article Title: Long-term aerobic exercise enhances circulating exosomal miR-214-3p to promote endothelial progenitor cell-mediated repair of endothelial damage induced by obesity

doi: 10.1016/j.jshs.2025.101094

Figure Lengend Snippet: miR-214-3p plays an essential role in exercise-mediated protection against obesity-induced EPC dysfunction in vivo . (A) Dynamic weight change curves of rats. After randomization at Week 10, rats underwent treadmill exercise until the end of Week 18 ( n : 5–7 for each group). * p < 0.05, KO + HE vs . WT + NC; ### p < 0.001, WT + HC vs . WT + NC; ††† p < 0.001, WT + HE vs . WT + NC; ‡‡‡ p < 0.001, WT + HE vs . WT + HC; §§ p < 0.01, KO + HE vs . WT + HE. (B) Summary data for Lee's index among groups at Week 10 and Week 18 ( n : 5–7 for each group). Left panel: ** p < 0.01, KO + HE vs . WT + NC; ### p < 0.001, WT + HC vs . WT + NC; ††† p < 0.001, WT + HE vs . WT + NC; Right panel: *** p < 0.001, KO + HE vs . WT + HC; ### p < 0.001, WT + HC vs . WT + NC; ‡‡‡ p < 0.001, WT + HE vs . WT + HC. (C) Summary data for acetylcholine (ACh)-induced, endothelium-dependent relaxation in mesenteric arteries among groups ( n : 5–6 for each group). * p < 0.05, WT + HE vs . KO + HE; # p < 0.05, ## p < 0.05, WT + HE vs . WT + HC. (D) Summary data for EC 50 values in mesenteric arteries among groups in response to ACh ( n : 5–6 for each group). * p < 0.05, KO + HE vs . WT + HE; ## p < 0.01, WT + HE vs . WT + HC; ‡‡ p < 0.01, WT + HC vs . WT + NC. (E) Cell proliferation assays demonstrated that exosomes derived from the WT + HC group exhibited a diminished capacity to promote EPC proliferation compared to the WT + NC group in rats. In contrast, exosomes derived from the WT + HE group significantly enhanced EPC proliferation, which was abolished by knocking out miR-214-3p in rats ( n = 10 for each group). *** p < 0.001, KO + HE vs . WT + HE; ### p < 0.001, WT + HE vs . WT + HC; ‡‡ p < 0.01, WT + HC vs . WT + NC. (F) Scratch assay results showed that exosomes derived from the WT + HC group exhibited a diminished capacity to enhance EPC migration rates compared to the WT + NC group in rats. In contrast, exosomes derived from the WT + HE group significantly enhanced EPC migration rates, which was abolished by knocking out miR-214-3p in rats ( n = 6 for each group). *** p < 0.001, KO + HE vs . WT + HE; ### p < 0.001, WT + HE vs . WT + HC; ‡‡ p < 0.01, WT + HC vs . WT + NC. (G) Representative images of wound healing in the scratch assay, showcasing the migratory response of rat EPC. (H) qPCR analyses of pre-miR-214-3p and miR-214-3p in tissues from obese rats with and without exercise training ( n : 4–6 for each group). *** p < 0.001, HE vs . HC. EC 50 = half maximal effective concentration; EPC = endothelial progenitor cells; HC = the high-fat diet with sedentary group; HE = the high-fat diet with exercise group; KO + HE = the knockout + high-fat diet with exercise group; miR = microRNA; pre-miR = precursor microRNA; WT + HC = the wild-type + high-fat diet with sedentary group; WT + HE = the wild-type + high-fat diet with exercise group; WT + NC = the wild-type + normal diet with sedentary group.

Article Snippet: Total RNAs from tissues, cells, and exosomes were extracted using Trizol (R0016; Beyotime Biotech, Shanghai, China), following the manufacturer’s recommendations. mRNA samples underwent reverse transcription using the Evo M-MLV Kit (AG11705; Accurate Biotechnology, Changsha, China), while miRNA samples were reversely transcribed using the All-in-OneTM miRNA qPCR Kit (QP115; iGene Biotechnology, Guangzhou, China), following the provided instructions.

Techniques: In Vivo, Derivative Assay, Wound Healing Assay, Migration, Concentration Assay, Knock-Out

Journal: Bioactive Materials

Article Title: Exercise-derived exosomal miR-151-3p: An innovative anti-inflammatory and antioxidant therapeutic for spinal cord injury

doi: 10.1016/j.bioactmat.2026.06.009

Figure Lengend Snippet: miR-151-3p is a key functional cargo in Exe-Exos mediating anti-apoptotic and antioxidant effects. (A – B) Differentially expressed miRNAs between Exe-Exos and Sed-Exos groups (q < 0.05, |log 2 FC| > 1). (C) RT-qPCR validation of 10 differentially expressed miRNAs. (D) Flow cytometry analysis of apoptosis in OGD/R, OGD/R + Exos, and OGD/R + Exos + miR-151-3p groups. (E) Quantification of apoptotic cell percentage (n = 3 independent cell culture experiments). (F) LDH activity (U/L; n = 5 independent cell culture experiments). (G) WB and (H – M) quantification of p-JNK, JNK, C-Caspase9, Caspase9, C-Caspase3, and Caspase3 protein expression (n = 6 independent biological replicates). Exos exosomes, JNK c-Jun N-terminal kinase.

Article Snippet: The miRNA stem-loop primers (Shanghai Jierui Bioengineering) are listed in .

Techniques: Functional Assay, Quantitative RT-PCR, Biomarker Discovery, Flow Cytometry, Cell Culture, Activity Assay, Expressing

Journal: Engineering Microbiology

Article Title: Cross-kingdom noncoding RNA regulation facilitates Nosema bombycis proliferation

doi: 10.1016/j.engmic.2026.100278

Figure Lengend Snippet: NbLNC2914 acts as a miRNA sponge that inhibits the expression of bmo-miR-2808a-3p.

Article Snippet: Next, the total RNAs were used for cDNA synthesis with the PrimeScriptTM RT reagent Kit (Takara, Japan) and miRNA first-strand cDNA synthesis (Tailing Reaction) (Sangon Biotech, China).

Techniques: Expressing

Journal: Oncology Letters

Article Title: Lectin-captured plasma microRNAs predict response and survival in patients with non-small cell lung cancer treated with nivolumab

doi: 10.3892/ol.2026.15669

Figure Lengend Snippet: Identification of circulating OAA1-captured miRNAs associated with lung cancer and resistance to nivolumab. (A) Lectin histochemistry using OAA1 on surgically resected lung cancer tissues (×200 magnification). The left panel shows OAA1 lectin staining in normal lung tissue, whereas the right panel shows OAA1 lectin staining in lung cancer tissue. (B) In the discovery phase, plasma samples from two patients with NSCLC with PD were collected before and after nivolumab treatment. After plasma purification using an OAA1 column, microarray analysis was performed to identify candidate miRNAs (miR-320a, miR-320b, and miR-3613-5p) associated with nivolumab resistance. In the validation phase, plasma samples from 48 patients with NSCLC (before nivolumab treatment) were analyzed with and without OAA1 column purification. The levels of the identified miRNAs were quantified using TaqMan qPCR, and statistical analyses were conducted to evaluate their predictive and prognostic values as biomarkers for nivolumab resistance. (C) Microarray analysis of OAA1-captured plasma miRNAs in patients with PD after nivolumab treatment reveals several upregulated candidate miRNAs, including miR-320a, miR-320b, and miR-3613-5p. (D) Relative levels of miR-320a, miR-320b, and miR-3613-5p in pre-treatment plasma from patients with NSCLC (n=48) and healthy controls (n=11) with and without OAA1 enrichment. Plasma miRNA levels were quantified by reverse transcription-qPCR. Relative miRNA levels were normalized to miR-16-5p and calculated using the 2 −ΔΔCq method, with the mean level of healthy volunteers serving as the calibrator. Statistical significance was determined using the Mann-Whitney U test. Data are presented as median with 95% confidence intervals. miRNA/miR, microRNA; NSCLC, non-small cell lung cancer; OAA1, Oscillatoria agardhii agglutinin 1; PD, progressive disease; qPCR, quantitative polymerase chain reaction.

Article Snippet: After thorough vortex mixing, the miRNA was purified using the NucleoSpin miRNA Plasma kit (Macherey-Nagel, Germany) with DNA digestion treatment, according to the manufacturer's protocol.

Techniques: Staining, Clinical Proteomics, Purification, Microarray, Biomarker Discovery, Reverse Transcription, MANN-WHITNEY, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Lectin-captured plasma microRNAs predict response and survival in patients with non-small cell lung cancer treated with nivolumab

doi: 10.3892/ol.2026.15669

Figure Lengend Snippet: Differential expression of miR-320a, miR-320b, and miR-3613-5p in pre-treatment plasma from patients with non-small cell lung cancer with PR or SD + PD. (A) Relative levels of miR-320a, miR-320b, and miR-3613-5p without OAA1 enrichment. (B) With OAA1 column enrichment. Plasma miRNA levels were quantified by reverse transcription-quantitative polymerase chain reaction and are shown as relative abundance using the 2 −ΔCq method normalized to miR-16-5p. Statistical significance was determined using the Mann-Whitney U test. Data are presented as median with 95% confidence intervals. miR, microRNA; OAA1, Oscillatoria agardhii agglutinin 1; SD, stable disease; PD, progressive disease; PR, partial response.

Article Snippet: After thorough vortex mixing, the miRNA was purified using the NucleoSpin miRNA Plasma kit (Macherey-Nagel, Germany) with DNA digestion treatment, according to the manufacturer's protocol.

Techniques: Quantitative Proteomics, Clinical Proteomics, Reverse Transcription, Real-time Polymerase Chain Reaction, MANN-WHITNEY

Journal: Oncology Letters

Article Title: Lectin-captured plasma microRNAs predict response and survival in patients with non-small cell lung cancer treated with nivolumab

doi: 10.3892/ol.2026.15669

Figure Lengend Snippet: ROC curve analysis of miRNAs for predicting non-response to immune checkpoint inhibitor therapy in patients with non-small cell lung cancer treated with nivolumab. (A) ROC curves for circulating miR-320a, miR-320b, and miR-3613-5p without OAA1 column enrichment. (B) With OAA1 enrichment. AUC, area under the curve; CI, confidence interval; miR, microRNA; OAA1, Oscillatoria agardhii agglutinin 1; ROC, receiver operating characteristic.

Article Snippet: After thorough vortex mixing, the miRNA was purified using the NucleoSpin miRNA Plasma kit (Macherey-Nagel, Germany) with DNA digestion treatment, according to the manufacturer's protocol.

Techniques:

Journal: Oncology Letters

Article Title: Lectin-captured plasma microRNAs predict response and survival in patients with non-small cell lung cancer treated with nivolumab

doi: 10.3892/ol.2026.15669

Figure Lengend Snippet: Kaplan-Meier analysis for OS based on pre-treatment plasma levels of miR-320a, miR-320b, and miR-3613-5p in patients with non-small cell lung cancer treated with nivolumab (n=48). (A) Kaplan-Meier curves comparing the OS between the high- and low-expression groups for each miRNA without OAA1 column enrichment. (B) Kaplan-Meier curves for the same miRNAs with the OAA1 enrichment. The number of patients in each group is indicated in the legends. Cutoff values for high/low expression were determined using receiver operating characteristic analysis. P-values are shown; *P<0.05. miR, microRNA; OAA1, Oscillatoria agardhii agglutinin 1; OS, overall survival.

Article Snippet: After thorough vortex mixing, the miRNA was purified using the NucleoSpin miRNA Plasma kit (Macherey-Nagel, Germany) with DNA digestion treatment, according to the manufacturer's protocol.

Techniques: Clinical Proteomics, Expressing

Journal: International Journal of Surgery (London, England)

Article Title: Therapeutic effects of Icariside II on radiation cystitis: revealing the mechanistic role of the FN1/Itgαvβ6-PI3K/AKT signaling pathway using single-cell RNA sequencing

doi: 10.1097/JS9.0000000000005212

Figure Lengend Snippet: ICA II attenuates radiation-induced cellular damage by disrupting FN1–Itgαvβ6 binding and regulating the PI3K-AKT signaling pathway in bladder cells. (A) Representative surface plasmon resonance (SPR) sensorgrams and kinetic fitting curves showing the binding of ICA II to Itgαvβ6. (B) SPR sensorgrams and kinetic fitting curves for the interaction between FN1 and Itgαvβ6. (C) SPR sensorgrams and kinetic fitting curves of FN1 binding to Itgαvβ6 in the presence of 10 μM ICA II. (D) SPR sensorgrams and kinetic fitting curves of FN1 binding to Itgαvβ6 in the presence of 100 μM ICA II. (E) Effects of different radiation doses on SV-HUC-1 cell proliferation. (F) Effects of different ICA II concentrations on SV-HUC-1 cell proliferation. (G) Effects of different radiation doses on human bladder fibroblast (HBF) cell proliferation. (H) Effects of different ICA II concentrations on HBF cell proliferation. (I) Effect of ICA II on the repair of radiation-induced damage in SV-HUC-1 cells, as shown by proliferation and morphological changes after treatment. (J) Inhibitory effect of ICA II on the proliferation of HBF cells caused by radiation damage, indicating reduced fibroblast proliferation after ICA II treatment. (K and L) Western blot analysis of the effect of ICA II on the protein expression of Itgαvβ6 in SV-HUC-1 cells; the results revealed a dose-dependent decrease in expression following ICA II treatment. (M and N) Western blot analysis of the effect of ICA II on the protein expression of FN1 in HBF cells, which revealed a significant reduction in FN1 expression after ICA II treatment. (O) Enzyme-Linked Immunosorbent Assay detection of FN1 expression levels in the culture supernatant of HBF cells, confirming the suppression of FN1 secretion in response to ICA II treatment. (P) RT‒qPCR analysis was used to determine the silencing efficiency of FN1-targeted siRNA (siFN1), which successfully knocked down FN1 expression at both the mRNA and protein levels. (Q) Co-immunoprecipitation showing the interaction between FN1 and Itgαv/Itgβ6 in SV-HUC-1 cells treated with fibroblast-conditioned media. (R and S) ICA II treatment reduced EMT and fibrosis-related protein expression and inhibited PI3K-AKT pathway activation in radiation-damaged cells, suggesting that ICA II plays a protective role through regulating FN1 and Itgαvβ6 interactions. Statistical significance is indicated using standard notation.

Article Snippet: HBF cells were transiently transfected with small interfering RNA (siRNA) targeting FN1 (MCE, HY-RS05013) via the siRNA Transfection Reagent (MCE, HY-K2017) according to the manufacturer’s instructions.

Techniques: Binding Assay, SPR Assay, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Immunoprecipitation, Activation Assay