Journal: BMC Biology

Article Title: Machine learning combined with omics-based approaches reveals T-lymphocyte cellular fate imbalance in abdominal aortic aneurysm

doi: 10.1186/s12915-025-02400-x

Figure Lengend Snippet: Identification of TIRS regulatory mechanisms and key biomarkers. A LASSO-based feature selection, with the optimal lambda determined when the partial likelihood deviance reached the minimum value (left). SVM-RFE-based feature selection, with root mean square error (RMSE) reached the minimum value and R -squared reached the max value (mid). Venn diagram presented the intersection of key biomarkers obtained through both algorithms (right). B Aberrant expression profiles for key biomarkers in Abdominal Aortic Wall Dataset 1 (AAA n = 80 patients, control n = 10 healthy individuals; Student’s t -test). C ROC curve demonstrating the diagnostic efficacy of FOSB, JUNB, CST7, and TBC1D4 in Abdominal Aortic Wall Dataset 1. D Clinical impact plot illustrating the clinical utility of key biomarkers. The “Number high risk” curve closely aligns with the “Number high risk with the event” curve at each threshold probability, indicating exceptional predictive power. E Aberrant expression profiles for key biomarkers in Abdominal Aortic Wall Dataset 2 (AAA n = 9 patients, control n = 10 healthy individuals; Student’s t -test). F ROC curve validating the diagnostic efficacy of FOSB, JUNB, CST7, and TBC1D4 in Abdominal Aortic Wall Dataset 2. G Clinical impact plot demonstrating the clinical utility of key biomarkers. Again, the “Number high risk” curve is closely aligned with the “Number high risk with the event” curve at each threshold probability, highlighting the biomarkers’ strong predictive power. H Aberrant expression profiles for key biomarkers in Perivascular Adipose Tissue Dataset 3 (dilated n = 30, non-dilated n = 30; Student’s t -test). I ROC curve verifying the diagnostic efficacy of FOSB, JUNB, CST7, and TBC1D4 in Perivascular Adipose Tissue Dataset 3. J Impact plots reiterated superior predictive performance probability, indicating outstanding predictive capability

Article Snippet: The primary antibodies against FOSB (1:500, catalog No. ab184938, Abcam) and JUNB (1:50, catalog No. 10486–1-AP, Proteintech) were incubated overnight at 4 °C, followed by incubation with horseradish peroxidase conjugated secondary antibodies.

Techniques: Selection, Expressing, Control, Diagnostic Assay

Journal: BMC Biology

Article Title: Machine learning combined with omics-based approaches reveals T-lymphocyte cellular fate imbalance in abdominal aortic aneurysm

doi: 10.1186/s12915-025-02400-x

Figure Lengend Snippet: Verification of key biomarkers. A Abdominal aortic wall and peripheral blood samples obtained from AAA patients. B Aberrant expression profiles for key biomarkers in the abdominal aortic wall (Inhouse Dataset 1; AAA n = 5 patients, control n = 4 healthy individuals). C ROC curve validating the diagnostic efficacy of FOSB, JUNB, CST7, and TBC1D4 in the abdominal aortic wall (Inhouse Dataset 1). D Clinical impact plot demonstrating the clinical utility of key biomarkers. The “Number high risk” curve closely aligns with the “Number high risk with the event” curve at each threshold probability, indicating exceptional predictive power. E Aberrant expression profiles for key biomarkers in peripheral blood (Inhouse Dataset 2; AAA n = 24 patients, control n = 15 healthy individuals). F ROC curve validating the diagnostic efficacy of FOSB, JUNB, CST7, and TBC1D4 in peripheral blood (Inhouse Dataset 2). G Clinical impact plot illustrating the clinical utility of key biomarkers. Again, the “Number high risk” curve remains closely aligned with the “Number high risk with the event” curve at each threshold probability, highlighting the biomarkers’ strong predictive capability. H Mice were infused with saline or Ang II (1000 ng/kg/min) + BAPN. Gross abdominal aorta images were shown. Scale bar is 1 cm. I Representative images of immunohistochemical stains for elastin fiber (Van Gieson) and representative photomicrographs of hematoxylin and eosin (H&E) staining. Scale bar is 200 μm. J – L Representative immunohistochemical staining of FOSB and JUNB in aortic cross sections. Scale bar is 50 μm. Data are expressed as mean ± SEM (control n = 3 mice, AAA n = 5 or 6 mice). Student’s t -test was utilized to compare continuous variables between the two groups

Article Snippet: The primary antibodies against FOSB (1:500, catalog No. ab184938, Abcam) and JUNB (1:50, catalog No. 10486–1-AP, Proteintech) were incubated overnight at 4 °C, followed by incubation with horseradish peroxidase conjugated secondary antibodies.

Techniques: Expressing, Control, Diagnostic Assay, Saline, Immunohistochemical staining, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Extracellular vesicle‐carried microRNA‐27b derived from mesenchymal stem cells accelerates cutaneous wound healing via E3 ubiquitin ligase ITCH

doi: 10.1111/jcmm.15692

Figure Lengend Snippet: Primer sequences for RT‐qPCR

Article Snippet: The antibodies used included ITCH (A15612, 1:2000, rabbit antibody, abclonal), JUNB (A5290, 1:2000, rabbit antibody, abclonal), IRE1α (ab124945, 1:10 000, rabbit antibody, Abcam), HRP‐labelled anti‐rabbit immunoglobulin G (IgG; 7074, 1:5000, CST) and anti‐mouse IgG (7076, 1:5000, CST).

Techniques: Sequencing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Extracellular vesicle‐carried microRNA‐27b derived from mesenchymal stem cells accelerates cutaneous wound healing via E3 ubiquitin ligase ITCH

doi: 10.1111/jcmm.15692

Figure Lengend Snippet: The delivery of mR‐27b from MSC‐derived EVs enhances the proliferation and migration of HaCaT cells and HSFs through JUNB/IRE1α up‐regulation by repressing ITCH both in vitro and in vivo. HaCaT cells and HSFs transfected with OE‐NC or OE‐ITCH used for following detections in panels A and B were co‐treated with PBS and MSC‐derived EVs, respectively. (A) Expression of miR‐27b, ITCH, JUNB and IRE1α was determined by RT‐qPCR in HaCaT cells and HSFs after different treatments, normalized to U6 and GAPDH, respectively. (B) Representative Western blots of ITCH, JUNB and IRE1α proteins and their quantitation in HaCaT cells and HSFs after different treatments, normalized to GAPDH. * P < .05 compared with HaCaT cells and HSFs transfected with OE‐NC and treated with PBS; # P < .05 compared with HaCaT cells and HSFs transfected with OE‐NC and treated with MSC‐derived EVs. HaCaT cells and HSFs used for following detections in panels C and D were transfected with OE‐NC or OE‐JUNB. (C) Relative expression of JUNB was determined by RT‐qPCR in cells after different treatments, normalized to GAPDH. (D) Representative Western blots of JUNB protein and its quantitation in cells after different treatments, normalized to GAPDH; * P < .05 compared with OE‐NC transfection. HaCaT cells and HSFs used for following detections in panels (E, G, H and I) were co‐transfected with OE‐ITCH and/or OE‐JUNB. (E) Representative Wester blots of ITCH, JUNB and IRE1α proteins and their quantitation in cells after different treatments, normalized to GAPDH. (F) Effect of OE‐ITCH on JUNB ubiquitination was determined by Co‐IP assay. (G) Proliferation of HaCaT cells and HSFs was examined by CCK‐8 assay after different treatments. (H) Migration of HaCaT cells and HSFs was examined by Transwell assay after different treatments (scale bar = 50 μm). (I) Migration rate of HaCaT cells and HSFs was examined by scratch test after different treatments; * P < .05 compared with OE‐NC treatment; # P < .05 compared with OE‐ITCH treatment. (J) Representative Western blots of ITCH, JUNB and IRE1α proteins and their quantitation in skin tissues of wounds in mice after treatments with PBS, MSC‐derived EVs and inhibitor‐NC, or MSC‐derived EVs and miR‐27b inhibitor, normalized to GAPDH. * P < .05 compared with PBS treatment. Data are shown as mean ± standard deviation of three technical replicates. Data between the two groups were compared by unpaired t test

Article Snippet: The antibodies used included ITCH (A15612, 1:2000, rabbit antibody, abclonal), JUNB (A5290, 1:2000, rabbit antibody, abclonal), IRE1α (ab124945, 1:10 000, rabbit antibody, Abcam), HRP‐labelled anti‐rabbit immunoglobulin G (IgG; 7074, 1:5000, CST) and anti‐mouse IgG (7076, 1:5000, CST).

Techniques: Derivative Assay, Migration, In Vitro, In Vivo, Transfection, Expressing, Quantitative RT-PCR, Western Blot, Quantitation Assay, Ubiquitin Proteomics, Co-Immunoprecipitation Assay, CCK-8 Assay, Transwell Assay, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Extracellular vesicle‐carried microRNA‐27b derived from mesenchymal stem cells accelerates cutaneous wound healing via E3 ubiquitin ligase ITCH

doi: 10.1111/jcmm.15692

Figure Lengend Snippet: MSC‐derived EVs mitigate epidermal re‐epithelialization and collagen fibre proliferation in wounds by delivering miR‐27b in vivo. The mice used for following detections were treated with PBS, EVs derived from MSCs transfected with inhibitor‐NC or EVs derived from MSCs transfected with miR‐27b‐inhibitor. (A) Size of wounds in mice at the 0, 2, 4, 6 and 8 d after operation. (B) Healing rate of wounds. (C) Pathological changes of cutaneous wound tissues were observed by HE staining in mice at 8 d after operation (scale bar = 250 μm). (D) Quantified scar width in the wound of mice. (E) Wound re‐epithelialization in mice. (F) Masson‐stained collagen in wound tissues of mice (scale bar = 250 μm). (G) miR‐27b expression was determined by RT‐qPCR in cutaneous wound tissues of mice at the 8 d after operation, normalized to U6. (H) Representative Western blots of ITCH, JUNB and IRE1α proteins and their quantitation in cutaneous wound tissues of mice, normalized to GAPDH. * P < .05 compared with PBS treatment; # P < .05 compared with EVs derived from MSCs transfected with inhibitor‐NC by unpaired t test. The data above were measurement data presented as mean ± standard deviation. n = 15 for mice following each treatment

Article Snippet: The antibodies used included ITCH (A15612, 1:2000, rabbit antibody, abclonal), JUNB (A5290, 1:2000, rabbit antibody, abclonal), IRE1α (ab124945, 1:10 000, rabbit antibody, Abcam), HRP‐labelled anti‐rabbit immunoglobulin G (IgG; 7074, 1:5000, CST) and anti‐mouse IgG (7076, 1:5000, CST).

Techniques: Derivative Assay, In Vivo, Transfection, Staining, Expressing, Quantitative RT-PCR, Western Blot, Quantitation Assay, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Extracellular vesicle‐carried microRNA‐27b derived from mesenchymal stem cells accelerates cutaneous wound healing via E3 ubiquitin ligase ITCH

doi: 10.1111/jcmm.15692

Figure Lengend Snippet: The molecular mechanism of MSC‐derived EVs carrying miR‐27b in affecting cutaneous wound healing through the ITCH/JUNB/IRE1α axis. MSC‐derived EVs carrying miR‐27b suppressed the ITCH expression in HaCaT cells and HSFs, and consequently enhanced the expression of JUNB, and IRE1α, thus improving the proliferation and migration of HaCaT cells and HSFs and accelerating the cutaneous wound healing in mice

Article Snippet: The antibodies used included ITCH (A15612, 1:2000, rabbit antibody, abclonal), JUNB (A5290, 1:2000, rabbit antibody, abclonal), IRE1α (ab124945, 1:10 000, rabbit antibody, Abcam), HRP‐labelled anti‐rabbit immunoglobulin G (IgG; 7074, 1:5000, CST) and anti‐mouse IgG (7076, 1:5000, CST).

Techniques: Derivative Assay, Expressing, Migration