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: iScience

Article Title: Construction of in vitro models derived from distinct regions of mouse nasal mucosa

doi: 10.1016/j.isci.2026.117611

Figure Lengend Snippet: Structural organization and cellular composition of the olfactory and respiratory regions in nasal mucosa (A) Architecture and cellular composition of the human olfactory epithelium (OE) and respiratory epithelium (RE). (B) Schematic diagram delineating the murine OE and RE regions, based on HE staining and immunostaining for olfactory sensory neurons (OSNs) markers OMP and Tuj1 (top: coronal views; bottom: horizontal views). (C) Representative immunostaining images of murine OE markers (OMP, Tuj1) and RE markers (β-Tubulin Ⅳ, MUC5AC). (D) Relative mRNA expression levels of representative marker genes analyzed by RT-qPCR. Nuclei were counterstained with Hoechst (blue). Scale bars, 50 μm (except for B, 500 μm). Data are presented as mean ± SD; ns: non-significant, ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: ChamQ Universal SYBR qPCR Master Mix , Vazyme , Cat# Q711.

Techniques: Olfactory, H&E Stain, Immunostaining, Expressing, Marker, Quantitative RT-PCR

Journal: iScience

Article Title: Construction of in vitro models derived from distinct regions of mouse nasal mucosa

doi: 10.1016/j.isci.2026.117611

Figure Lengend Snippet: Culture and characterization of primary cells from RE and OE regions (A) Schematic depicting the outward radial expansion of primary cells from a single cell origin. (B) Phase-contrast images of RE- and OE-derived cells after 5 days of culture. (C) Immunostaining for β-Tubulin Ⅳ and ZO-1 in expanded RE- and OE-derived cultures, the rightmost image shows enlarged views of the boxed areas, revealing cell undergoing mitosis marked by β-Tubulin Ⅳ. (D) Immunostaining for KRT5, TRP63, E-cadherin, and Ki67 in expanded RE- and OE-derived cultures. (E) Quantification of specific cell numbers per field of view in RE- and OE-derived cultures. (F) Relative mRNA expression levels of basal epithelial markers in RE- and OE-derived cultures analyzed by RT-qPCR. Nuclei were counterstained with Hoechst (blue). Scale bars, 50 μm. Data are presented as mean ± SD; ns: non-significant, ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: ChamQ Universal SYBR qPCR Master Mix , Vazyme , Cat# Q711.

Techniques: Single Cell, Phase Contrast, Derivative Assay, Immunostaining, Expressing, Quantitative RT-PCR

Journal: iScience

Article Title: Construction of in vitro models derived from distinct regions of mouse nasal mucosa

doi: 10.1016/j.isci.2026.117611

Figure Lengend Snippet: IL-17a inhibits ciliary differentiation in RE-ALI model (A) HE staining of longitudinal sections from RE-ALI cultures treated with IL-17a, IL-17a + Nab, or CHIR99021. (B) Proportion of ciliary region length per 500 μm ALI cultures. (C) Immunostaining of β-Tubulin IV, MUC5AC, and Ace-tubulin in RE-ALI cultures under the indicated treatment conditions. (D) Quantification of β-Tubulin IV + clusters, MUC5AC + cells or Ace-Tubulin + clusters per field of view. (E) Relative mRNA expression levels of Foxj1, Tubb4a, or Muc5ac in RE-ALI cultures analyzed by RT-qPCR. Nuclei were counterstained with Hoechst (blue). Scale bars, 50 μm. Data are presented as mean ± SD, ns: non-significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: ChamQ Universal SYBR qPCR Master Mix , Vazyme , Cat# Q711.

Techniques: H&E Stain, Immunostaining, Expressing, Quantitative RT-PCR

Journal: iScience

Article Title: Construction of in vitro models derived from distinct regions of mouse nasal mucosa

doi: 10.1016/j.isci.2026.117611

Figure Lengend Snippet:

Article Snippet: ChamQ Universal SYBR qPCR Master Mix , Vazyme , Cat# Q711.

Techniques: Recombinant, Tissue, Purification, Software

Journal: iScience

Article Title: A multimodal in situ framework for spatial characterization applied on SARS-CoV-2 live-attenuated vaccines in the nasal mucosa

doi: 10.1016/j.isci.2026.117730

Figure Lengend Snippet:

Article Snippet: qScript XLT One-Step RT-qPCR ToughMix , QuantaBio , Cat# 95132–500.

Techniques: Plasmid Preparation, Virus, Indirect ELISA, Recombinant, RNAscope, Multiplex Assay, Software, Negative Control, Confocal, Laser-Scanning Microscopy

Journal: iScience

Article Title: Lateralized vagal oxytocin receptor signaling separately controls feeding and socioemotional functions via hypothalamic oxytocin neurons

doi: 10.1016/j.isci.2026.117713

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Article Snippet: ReverTra Ace qPCR RT Master Mix , Toyobo , Cat# FSQ-201.

Techniques: Cell-Signaling, Virus, Recombinant, Software, Imaging, Fluorescence, Real-time Polymerase Chain Reaction

Journal: iScience

Article Title: A method for expressing two proteins at a tunable ratio allows for studying the kinetics of creatine transport by MCT12

doi: 10.1016/j.isci.2026.117719

Figure Lengend Snippet:

Article Snippet: Luna Universal One-Step RT-qPCR Kit , NEB , #E3005.

Techniques: Cell-Signaling, Virus, Recombinant, Clinical Proteomics, Membrane, Staining, Cloning, Software

Journal: iScience

Article Title: Lateralized vagal oxytocin receptor signaling separately controls feeding and socioemotional functions via hypothalamic oxytocin neurons

doi: 10.1016/j.isci.2026.117713

Figure Lengend Snippet:

Article Snippet: THUNDERBIRD NEXT SYBR qPCR Mix , Toyobo , Cat# QPS-201.

Techniques: Cell-Signaling, Virus, Recombinant, Software, Imaging, Fluorescence, Real-time Polymerase Chain Reaction