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Role of PVPAC-Exo-circEif3c in regulating AF biological functions and its potential mechanism. PVPAC-derived exosomal circEif3c (Exo-circEif3c) promoted AFs migration and proliferation, whereas silencing exosomal circEif3c suppresses these processes. (A) Time-course analysis of circEif3c expression in AFs after Exo-circEif3c treatment (0, 6, and 12 h; 0 h as control). (B) Stable silencing efficiency and specificity of circEif3c in AFs; Exo-siR-control served as the control. (C and D) Effects of PVPAC-Exo-siR- circEif3c-1 and -2 on AF migration and proliferation assessed by wound healing and proliferation assays. Scratch closure percentage and migrated cell numbers were quantified using ImageJ and GraphPad Prism 9.5, scale bar = 150 μm. (E) and (F) FCM analysis of AF proliferation and apoptosis following treatment with PVPAC-Exo-circEif3c, Exo-miR-96–5p, and <t>Ad-MEOX2</t> interaction. (G) Western blot analysis of vimentin, PHF20L1, and MEOX2 expression in AFs under high glucose and circEif3c modulation. (H) Effects of Exo-circEif3c on the expression of vimentin, PHF20L1, MEOX2, and LC3 in AFs. GAPDH was used as a loading control. All data above are presented as mean ± SD from three independent experiments. vs. the control group, ∗P < 0.05, ∗∗P < 0.01(one-way ANOVA with Dunnett's post-hoc test), n (the number of experiments) = 3.
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Schematic illustration of the preparation of CPs@SS31, and the corresponding in vivo therapy of acute lung injury via NIR enhanced ROS scavenging, inflammation inhibition, macrophage M2 polarization, and T cells immunoactivation, as well as specifically targeting mitochondria, activating mitochondrial function, and inducing mitophagy to reprogram lung redox homeostasis, and promote tissue repair.

Journal: Bioactive Materials

Article Title: Near infrared enhanced palladium loaded siraitia grosvenorii carbon dots amplify mitophagy for acute lung injury immunotherapy

doi: 10.1016/j.bioactmat.2026.02.040

Figure Lengend Snippet: Schematic illustration of the preparation of CPs@SS31, and the corresponding in vivo therapy of acute lung injury via NIR enhanced ROS scavenging, inflammation inhibition, macrophage M2 polarization, and T cells immunoactivation, as well as specifically targeting mitochondria, activating mitochondrial function, and inducing mitophagy to reprogram lung redox homeostasis, and promote tissue repair.

Article Snippet: Cell viability testing : Mouse monocyte macrophages (RAW264.7, ATCC, USA) were cultured in DMEM containing 10% FBS and 1% penicillin-streptomycin (Solarbio, China).

Techniques: In Vivo, Inhibition

Role of PVPAC-Exo-circEif3c in regulating AF biological functions and its potential mechanism. PVPAC-derived exosomal circEif3c (Exo-circEif3c) promoted AFs migration and proliferation, whereas silencing exosomal circEif3c suppresses these processes. (A) Time-course analysis of circEif3c expression in AFs after Exo-circEif3c treatment (0, 6, and 12 h; 0 h as control). (B) Stable silencing efficiency and specificity of circEif3c in AFs; Exo-siR-control served as the control. (C and D) Effects of PVPAC-Exo-siR- circEif3c-1 and -2 on AF migration and proliferation assessed by wound healing and proliferation assays. Scratch closure percentage and migrated cell numbers were quantified using ImageJ and GraphPad Prism 9.5, scale bar = 150 μm. (E) and (F) FCM analysis of AF proliferation and apoptosis following treatment with PVPAC-Exo-circEif3c, Exo-miR-96–5p, and Ad-MEOX2 interaction. (G) Western blot analysis of vimentin, PHF20L1, and MEOX2 expression in AFs under high glucose and circEif3c modulation. (H) Effects of Exo-circEif3c on the expression of vimentin, PHF20L1, MEOX2, and LC3 in AFs. GAPDH was used as a loading control. All data above are presented as mean ± SD from three independent experiments. vs. the control group, ∗P < 0.05, ∗∗P < 0.01(one-way ANOVA with Dunnett's post-hoc test), n (the number of experiments) = 3.

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: Role of PVPAC-Exo-circEif3c in regulating AF biological functions and its potential mechanism. PVPAC-derived exosomal circEif3c (Exo-circEif3c) promoted AFs migration and proliferation, whereas silencing exosomal circEif3c suppresses these processes. (A) Time-course analysis of circEif3c expression in AFs after Exo-circEif3c treatment (0, 6, and 12 h; 0 h as control). (B) Stable silencing efficiency and specificity of circEif3c in AFs; Exo-siR-control served as the control. (C and D) Effects of PVPAC-Exo-siR- circEif3c-1 and -2 on AF migration and proliferation assessed by wound healing and proliferation assays. Scratch closure percentage and migrated cell numbers were quantified using ImageJ and GraphPad Prism 9.5, scale bar = 150 μm. (E) and (F) FCM analysis of AF proliferation and apoptosis following treatment with PVPAC-Exo-circEif3c, Exo-miR-96–5p, and Ad-MEOX2 interaction. (G) Western blot analysis of vimentin, PHF20L1, and MEOX2 expression in AFs under high glucose and circEif3c modulation. (H) Effects of Exo-circEif3c on the expression of vimentin, PHF20L1, MEOX2, and LC3 in AFs. GAPDH was used as a loading control. All data above are presented as mean ± SD from three independent experiments. vs. the control group, ∗P < 0.05, ∗∗P < 0.01(one-way ANOVA with Dunnett's post-hoc test), n (the number of experiments) = 3.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Derivative Assay, Migration, Expressing, Control, Western Blot

The regulatory mchanisms of miR-96-5p in AF biology. (A) Time-course analysis of miR-96–5p expression in AFs treated with PVPAC-Exo at 0, 6, 12, and 24 h. (B) and (C) AFs were transfected with miR-96–5p mimic and NC mimic for 24 h. Then, the migration ability of AFs through migration experiments (B) and EDU assay (C) were evaluated using Image J and GraphPad Prism 9. vs. control mimic, ∗P < 0.05, ∗∗P < 0.01, n (the number of experiments) = 3, scale bar = 150 μm. (D) Correlation analysis between extracellular ncRNA levels in culture supernatant and intracellular expression of PHF20L1 and MEOX2 by RT-qPCR. (E) Bioinformatic prediction identifying PHF20L1 and MEOX2 as potential targets of miR-96–5p. (F) Predicted miR-96–5p binding sites in the 3′UTR of PHF20L1. (G) Luciferase reporter assay displayed that miR-96–5p mimic significantly reduced luciferase activity in the PHF20L1-WT group, but not in the PHF20L1-Mut group (vs. PHF20L1-Mut, ∗∗ P < 0.01), n (the number of experiments) = 3. (H) Bioinformatics analysis indicated the miR-96–5p binding sites in the 3′UTR of MEOX2. (I) Western blot exhibited no significant change in MEOX2 protein levels upon miR-96–5p overexpression. (J) Interaction network among miR-96–5p, circEif3c, PHF20L1, and MEOX2, constructed using GEPIA, ENCORI, miRNet, NDEx, and Cytoscape. (K) Western blot analysis of PHF20L1 and MEOX2 expression in AFs transfected with control-exosome, OE-exosomes, miR-comtrol mimic, miR-96–5p mimic, and UNC1215, respectively. vs. control-exosome, miR-comtrol mimic, ∗ P < 0.05, ∗∗ P < 0.01, n (the number of experiments) = 3. (L) Predicted protein–protein interaction interface between PHF20L1 and MEOX2 using Zdock 3.0.2 and PyMOL 2.5.5. (M) Co-IP experiments confirmed an interaction between PHF20L1 and MEOX2.

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: The regulatory mchanisms of miR-96-5p in AF biology. (A) Time-course analysis of miR-96–5p expression in AFs treated with PVPAC-Exo at 0, 6, 12, and 24 h. (B) and (C) AFs were transfected with miR-96–5p mimic and NC mimic for 24 h. Then, the migration ability of AFs through migration experiments (B) and EDU assay (C) were evaluated using Image J and GraphPad Prism 9. vs. control mimic, ∗P < 0.05, ∗∗P < 0.01, n (the number of experiments) = 3, scale bar = 150 μm. (D) Correlation analysis between extracellular ncRNA levels in culture supernatant and intracellular expression of PHF20L1 and MEOX2 by RT-qPCR. (E) Bioinformatic prediction identifying PHF20L1 and MEOX2 as potential targets of miR-96–5p. (F) Predicted miR-96–5p binding sites in the 3′UTR of PHF20L1. (G) Luciferase reporter assay displayed that miR-96–5p mimic significantly reduced luciferase activity in the PHF20L1-WT group, but not in the PHF20L1-Mut group (vs. PHF20L1-Mut, ∗∗ P < 0.01), n (the number of experiments) = 3. (H) Bioinformatics analysis indicated the miR-96–5p binding sites in the 3′UTR of MEOX2. (I) Western blot exhibited no significant change in MEOX2 protein levels upon miR-96–5p overexpression. (J) Interaction network among miR-96–5p, circEif3c, PHF20L1, and MEOX2, constructed using GEPIA, ENCORI, miRNet, NDEx, and Cytoscape. (K) Western blot analysis of PHF20L1 and MEOX2 expression in AFs transfected with control-exosome, OE-exosomes, miR-comtrol mimic, miR-96–5p mimic, and UNC1215, respectively. vs. control-exosome, miR-comtrol mimic, ∗ P < 0.05, ∗∗ P < 0.01, n (the number of experiments) = 3. (L) Predicted protein–protein interaction interface between PHF20L1 and MEOX2 using Zdock 3.0.2 and PyMOL 2.5.5. (M) Co-IP experiments confirmed an interaction between PHF20L1 and MEOX2.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Expressing, Transfection, Migration, EdU Assay, Control, Quantitative RT-PCR, Binding Assay, Luciferase, Reporter Assay, Activity Assay, Western Blot, Over Expression, Construct, Co-Immunoprecipitation Assay

CircEif3c modulates AF proliferation and migration via the miR-96-5p/PHF20L 1 /MEOX2 axis. (A–C) Cell migration and proliferation assays. AFs were transfected for 24 h with Ad-GFP, siR-circEif3c, miR-96–5p mimic, or siR-MEOX2. Migration (A) and proliferation (B) were quantified (C). (D–F) AFs were co-incubated for 48 h with control mimic, Exo-(siR-)circEif3c mimic, Exo-(siR-)miR-96–5p mimic, PVPAC-exosome (Exo-control), GW4869, or Exo-siR-pAd-MEOX2. Migration (D) and proliferation (E) were assessed (F), scale bar = 150 μm. (G) Cellular fluorescence immunolocalization. nuclei (DAPI, blue), circEif3c (Cy5, red), miR-96–5p (Cy3, orange-yellow), MEOX2 (GFP, green).Scale bar = 30 μm. The above data were presented as mean ± SD. vs. Ad-GFP group, ∗ P < 0.05, ∗∗ P < 0.01, n (the number of experiments) = 3.

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: CircEif3c modulates AF proliferation and migration via the miR-96-5p/PHF20L 1 /MEOX2 axis. (A–C) Cell migration and proliferation assays. AFs were transfected for 24 h with Ad-GFP, siR-circEif3c, miR-96–5p mimic, or siR-MEOX2. Migration (A) and proliferation (B) were quantified (C). (D–F) AFs were co-incubated for 48 h with control mimic, Exo-(siR-)circEif3c mimic, Exo-(siR-)miR-96–5p mimic, PVPAC-exosome (Exo-control), GW4869, or Exo-siR-pAd-MEOX2. Migration (D) and proliferation (E) were assessed (F), scale bar = 150 μm. (G) Cellular fluorescence immunolocalization. nuclei (DAPI, blue), circEif3c (Cy5, red), miR-96–5p (Cy3, orange-yellow), MEOX2 (GFP, green).Scale bar = 30 μm. The above data were presented as mean ± SD. vs. Ad-GFP group, ∗ P < 0.05, ∗∗ P < 0.01, n (the number of experiments) = 3.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Migration, Transfection, Incubation, Control, Fluorescence

Exosomal circEif3c/miR-96-5p/PHF20L1/MEOX2 axis drives vascular remodeling in vivo. (A) Workflow: a stable PVPAC line over-expressing circEif3c supplied exosomes (Exo-Ad-circEif3c, 10 μg/mouse) that were micro-injected into perivascular adipose tissue (PVAT) surrounding the left carotid artery for 4 weeks to initiate remodeling. Subsequently, after the model was established, treatments with (Exo)-Ad-GFP, (Exo)-Ad- circEif3c, (Exo)-Ad-miR-96–5p, and (Exo)-Ad-Meox2 were administered continuously for 2 weeks, respectively. Normal saline (NS) was used as a negative control. (B) Representative H&E-stained cross-sections and concomitant ultrasonography of the common carotid artery. Black scale bars = 50 μm, yellow scale bars = 1 mm, and white scale bars = 0.1 s. (C) Immunohistochemistry. Scale bars = 20 μm. (D) Western blotting. (E) Quantification of protein levels. (F) Tissue localization of Cy5-labeled circEif3c by immunofluorescence, scale bar = 100 μm. (G) Fluorescence intensity quantification. (H) Comparative fluorescence imaging of vascular sections: (H1) Bright-field H&E vs. dark-field GFP before and after Ad-MEOX2 transfection; Scale bars = 50 μm; (H2) DM-remodeling vs MEOX2-intervention groups. Scale bars = 30 μm. (I) Whole-animal in vivo imaging of Cy5 signal. All quantitative data above are presented as mean ± SD. vs. control, ∗ P < 0.01.∗∗ P < 0.01. n (the number of animals) = 6 in each group.

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: Exosomal circEif3c/miR-96-5p/PHF20L1/MEOX2 axis drives vascular remodeling in vivo. (A) Workflow: a stable PVPAC line over-expressing circEif3c supplied exosomes (Exo-Ad-circEif3c, 10 μg/mouse) that were micro-injected into perivascular adipose tissue (PVAT) surrounding the left carotid artery for 4 weeks to initiate remodeling. Subsequently, after the model was established, treatments with (Exo)-Ad-GFP, (Exo)-Ad- circEif3c, (Exo)-Ad-miR-96–5p, and (Exo)-Ad-Meox2 were administered continuously for 2 weeks, respectively. Normal saline (NS) was used as a negative control. (B) Representative H&E-stained cross-sections and concomitant ultrasonography of the common carotid artery. Black scale bars = 50 μm, yellow scale bars = 1 mm, and white scale bars = 0.1 s. (C) Immunohistochemistry. Scale bars = 20 μm. (D) Western blotting. (E) Quantification of protein levels. (F) Tissue localization of Cy5-labeled circEif3c by immunofluorescence, scale bar = 100 μm. (G) Fluorescence intensity quantification. (H) Comparative fluorescence imaging of vascular sections: (H1) Bright-field H&E vs. dark-field GFP before and after Ad-MEOX2 transfection; Scale bars = 50 μm; (H2) DM-remodeling vs MEOX2-intervention groups. Scale bars = 30 μm. (I) Whole-animal in vivo imaging of Cy5 signal. All quantitative data above are presented as mean ± SD. vs. control, ∗ P < 0.01.∗∗ P < 0.01. n (the number of animals) = 6 in each group.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques: In Vivo, Expressing, Injection, Saline, Negative Control, Staining, Immunohistochemistry, Western Blot, Labeling, Immunofluorescence, Fluorescence, Imaging, Transfection, In Vivo Imaging, Control

Schematic illustration of the PVPAC-Exo mediated circEif3c/miR-96–5p/PHF20L1/MEOX2 axis regulating vascular remodeling.

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: Schematic illustration of the PVPAC-Exo mediated circEif3c/miR-96–5p/PHF20L1/MEOX2 axis regulating vascular remodeling.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques: