Journal: Bioactive Materials

Article Title: Engineering a halloysite nanotube-enhanced hydrogel 3D skin model for modulated inflammation and accelerated wound healing

doi: 10.1016/j.bioactmat.2024.11.013

Figure Lengend Snippet: Viability and functionality assessment of HFFs in HNT-modified hydrogel (a) Representative images and percentage of HFFs positive for vimentin by IF staining on days 0, 3, and 7 in natural hydrogel and 1 μg/mL HNT-based hydrogel. Data are represented as mean ± SD. ∗ P < 0.05 (two-way ANOVA analysis). Scale bar = 20 μm. (b) Relative expression levels of talin2, plasminogen, fibronectin, and paxillin assessed by RT-PCR tests in cultured HFFs on day 3. Each dot represents one independent experiment. Data are represented as mean ± SD. ns, not significant, ∗ P < 0.05; ∗∗ P < 0.01 (unpaired Student's t-test). (c) Representative images and percentage of HFFs positive for vimentin by IF staining in natural hydrogel and 1 μg/mL HNT-based hydrogel. The cells were stimulated with cytokines (TNF-α, IL-17A, and IL-22) in different concentrations (0, 25, 50, 100 ng/mL). Scale bar = 100 μm. (d) Representative 3D images from the top view of HFFs positive for vimentin in I collagen-based hydrogel, natural hydrogel, and 1 μg/mL HNT-based hydrogel on days 1, 3 and 7. The cells were stimulated without or with cytokines (TNF-α, IL-17A, and IL-22, 50 ng/mL for each cytokine). Scale bar = 100 μm. (e) Relative expression levels of vimentin in HFFs in I collagen-based hydrogel, natural hydrogel, and 1 μg/mL HNT-based hydrogel on days 1, 3, and 7 without cytokine stimulation. Data are represented as mean ± SD. ∗ P < 0.05 (two-way ANOVA analysis). (f) Relative expression levels of vimentin in HFFs in I collagen-based hydrogel, natural hydrogel, and 1 μg/mL HNT-based hydrogel on days 1, 3, and 7 with cytokine (TNF-α, IL-17A, and IL-22, 50 ng/mL for each) stimulation. Data are represented as mean ± SD. ∗∗ P < 0.01 (two-way ANOVA analysis).

Article Snippet: Subsequently, the samples were incubated overnight at 4 °C with primary antibodies, including rabbit anti-Ki-67 (catalog #34330, CST, USA), keratin (K) 5 (catalog # 71536, CST, USA), vimentin (catalog # 10366-1-AP, Proteintech, China), fibronectin (catalog # 15613-1-AP, Proteintech, China), K17 (catalog # 17516-1-AP, Proteintech, China), mouse anti-K10 (catalog # ab9025, Abcam, UK), CD34 (catalog # 31120-1-AP, Proteintech, China), or IL-17A (catalog # 66148-1-Ig, Proteintech, China).

Techniques: Modification, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction, Cell Culture

Journal: Bioactive Materials

Article Title: Engineering a halloysite nanotube-enhanced hydrogel 3D skin model for modulated inflammation and accelerated wound healing

doi: 10.1016/j.bioactmat.2024.11.013

Figure Lengend Snippet: Inflammatory bioactivity of modified hydrogel and its potential for cell proliferation, migration, and adhesion. (a) Schematic of RNA-seq of the HFFs in hydrogel group. Four groups were set as group A (without HNT), group B (with HNT of 1 μg/mL), group C (without HNT, stimulated with TNF-α, IL-17A, and IL-22, 50 ng/mL) and group D (with HNT of 1 μg/mL, stimulated with with TNF-α, IL-17A, and IL-22, 50 ng/mL). (b) Heatmap of relative expression levels of genes associated with inflammation, proliferation and migration in group B (with HNT) compared to group A (without HNT). (c) KEGG pathway analysis of enriched genes showing the relevant functional terms in group D compared to group B. (d) Specific inflammatory clusters of genes related to KEGG analysis showing the relevant functional terms in group D compared to group B. (e) KEGG pathway analysis of enriched genes showing the relevant functional terms in group A compared to group C as well as group D compared to group B. (f) Venn diagram of differentially expressed genes overlapped in the four groups. (g) Representative 3D images of K17 expression in upper-layer from the top and side-view of the modified 3D skin model. (h) Relative expression levels of K17 in 0 μg/mL and 1 μg/mL HNT-based hydrogel with cytokine (TNF-α, IL-17A, and IL-22, 50 ng/mL) stimulation. Each dot represents one independent experiment. Data are represented as mean ± SD. ∗ P < 0.05 (unpaired student's t-test).

Article Snippet: Subsequently, the samples were incubated overnight at 4 °C with primary antibodies, including rabbit anti-Ki-67 (catalog #34330, CST, USA), keratin (K) 5 (catalog # 71536, CST, USA), vimentin (catalog # 10366-1-AP, Proteintech, China), fibronectin (catalog # 15613-1-AP, Proteintech, China), K17 (catalog # 17516-1-AP, Proteintech, China), mouse anti-K10 (catalog # ab9025, Abcam, UK), CD34 (catalog # 31120-1-AP, Proteintech, China), or IL-17A (catalog # 66148-1-Ig, Proteintech, China).

Techniques: Modification, Migration, RNA Sequencing, Expressing, Functional Assay

Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: HSD promoted Th17 cell development and worsened prostate inflammation in EAP mice. a Simple schematic diagram of the implementation plan of the experiment. b HE staining and inflammation scores of prostate tissues (scale bar = 100 μm, n = 4). c Tactile allodynia development in the 4 groups of mice ( n = 4). d The levels of IL-1β, TNF-α, IL-17A, and IFN-γ in the serum of the mice in the 4 groups ( n = 4). e Proportion of Th1 and Th17 cells of CD4 + T cells detected by flow cytometry in the splenic lymphocytes of vaccinated mice from those 4 groups ( n = 4). f The infiltration of Th17 cells in prostate tissues from mice was determined by immunofluorescence (white arrowheads, scale bar = 100 μm, n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. BF breeding feed, EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, IL-1β interleukin-1β, TNF-α tumor necrosis factor-α, IFN-γ interferon-γ, IL-17A interleukin-17A, DAPI 4',6-diamidino-2-phenylindole, Th T helper

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: Staining, Flow Cytometry, Immunofluorescence

Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: Transplantation of fecal microbiota from EAP mice with HSD elicited more severe prostate inflammation in psuedogerm-free mice. a Simple schematic diagram of the experimental workflow. b HE staining and inflammation scores of prostate tissues (scale bar = 100 μm, n = 4). c Development of tactile allodynia in mice belonging to the 4 groups ( n = 4). d The concentrations of IL-1β, TNF-α, IL-17A, and 5-HIAA in the serum of NOD mice in the 4 groups were measured ( n = 4). e Proportion of Th17 of CD4 + T cells, as determined by flow cytometry, among the splenic lymphocytes of immunized mice in the 4 groups ( n = 4). f The infiltration of Th17 cells in prostate tissues from mice was determined by immunofluorescence (white arrowheads, scale bar = 100 μm, n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. NOD nonobese male diabetic/LtJ, EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, FMT fecal microbiota transplantation, HE hematoxylin–eosin, IL-1β interleukin-1β, TNF-α tumor necrosis factor-α, IL-17A interleukin-17A, 5-HIAA 5-hydroxyindole acetic acid, DAPI 4',6-diamidino-2-phenylindole

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: Transplantation Assay, Staining, Flow Cytometry, Immunofluorescence

Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: 5-HIAA supplementation ameliorated the symptoms of EAP caused by HSD. a The levels of 5-HIAA secreted into the serum of the mice were measured in 4 groups: Ctrl + NSD, Ctrl + HSD, EAP + NSD, and EAP + HSD groups ( n = 4). b Simple schematic diagram of the experimental workflow. c HE staining and inflammation scores of prostate tissues (scale bar = 100 μm, n = 4). d Tactile allodynia development in NOD mice in the 4 groups ( n = 4). e Secretion levels of IL-1β, TNF-α, and IL-17A in the serum of mice in the 5 groups ( n = 4). f Flow cytometry was used to determine the proportion of Th17 of CD4 + T cells among the splenic lymphocytes of immunized mice in the 5 groups ( n = 4). g The infiltration of Th17 cells in prostate tissues from mice was determined by immunofluorescence (white arrowheads, scale bar = 100 μm, n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. NOD nonobese male diabetic/LtJ, BF breeding feed, EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, HE hematoxylin–eosin, IL-1β interleukin-1β, TNF-α tumor necrosis factor-α, IL-17A interleukin-17A, 5-HIAA 5-hydroxyindole acetic acid, DAPI 4',6-diamidino-2-phenylindole

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: Staining, Flow Cytometry, Immunofluorescence

Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: Inhibition of AHR abrogated the protective effects of 5-HIAA supplementation on EAP mice fed an HSD. a AHR levels in prostate tissues of mice among the Ctrl + NSD, Ctrl + HSD, EAP + NSD, EAP + HSD, and EAP + HSD + 5-HIAA groups ( n = 4). b A simple schematic diagram of the experimental workflow. c HE staining and inflammation scores of prostate tissues (scale bar = 100 μm, n = 4). d Tactile allodynia development in the mice from the Ctrl + NSD, Ctrl + HSD, EAP + NSD, EAP + HSD, EAP + HSD + 5-HIAA, and EAP + HSD + 5-HIAA + CH groups ( n = 4). e The levels of IL-1β, TNF-α, and IL-17A in the serum of the mice in the 6 groups ( n = 4). f Proportion of Th17 of CD4 + T cells, as determined by flow cytometry, among the splenic lymphocytes of immunized mice in the 6 groups (white arrowheads, n = 4). g The infiltration of Th17 cells in prostate tissues from mice was evaluated by immunofluorescence (scale bar = 100 μm, n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. NOD nonobese male diabetic/LtJ, BF breeding feed, EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, CH CH223191 (an AHR inhibitor), HE hematoxylin–eosin, IL-1β interleukin-1β, TNF-α tumor necrosis factor-α, IL-17A interleukin-17A, 5-HIAA 5-hydroxyindole acetic acid, AHR aryl hydrocarbon receptor, DAPI 4',6-diamidino-2-phenylindole

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: Inhibition, Staining, Flow Cytometry, Immunofluorescence

Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: HSD promoted the differentiation of Th17 cells by activating SGK1/FOXO1 signaling pathway. a The in vitro Th17 cell differentiation ratio was determined by flow cytometry ( n = 3). b Th17 cell differentiation ratio was determined in an in vitro differentiation experiment using immunofluorescence (scale bar = 100 μm, n = 3). c SGK1, p-FOXO1, and FOXO1 levels in prostate tissues from Ctrl + NSD, Ctrl + HSD, EAP + NSD, EAP + HSD, and EAP + HSD + 5-HIAA mice ( n = 4). d AHR, SGK1, p-FOXO1, and FOXO1 levels in prostate tissues of mice in the EAP + HSD + 5-HIAA and EAP + HSD + 5-HIAA + CH groups ( n = 4). e AHR, SGK1, p-FOXO1, and FOXO1 levels in naïve CD4 + T cells of Ctrl, Sti, Sti + HSD, Sti + HSD + 5-HIAA, Sti + HSD + 5-HIAA + CH groups ( n = 3). f Transcriptional levels of Sgk1 in naïve CD4 + T cells of Ctrl, Sti, Sti + HSD, Sti + HSD + 5-HIAA, Sti + HSD + 5-HIAA + CH groups (n = 3). g For the ChIP-qPCR assay, 3 potential enhancer segments of the mouse promoter region of Sgk1 were created. h qPCR was used to assess the immunoprecipitated DNA by a ChIP assay ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, IL-17A interleukin-17A, 5-HIAA 5-hydroxyindole acetic acid, AHR aryl hydrocarbon receptor, SGK1 serum and glucocorticoid-regulated kinase 1, FOXO1 forkhead box protein O1, CH CH223191 (an AHR inhibitor), ChIP chromatin immunoprecipitation, qPCR quantitative polymerase chain reaction, DAPI 4’,6-diamidino-2-phenylindole, Sti stimulation with IL-6, IL-23, TGF-β, anti-IFN-γ, and anti-IL-4

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: In Vitro, Cell Differentiation, Flow Cytometry, Immunofluorescence, ChIP-qPCR, Immunoprecipitation, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction

Journal: Frontiers in Pharmacology

Article Title: Licoisoflavone B alleviates psoriasis via SCD1-targeted lipid metabolism reprogramming and suppression of Th17/IL-17–mediated inflammation

doi: 10.3389/fphar.2026.1754729

Figure Lengend Snippet: Integrated workflow for target prediction and experimental validation of Lico B in psoriasis. Bioinformatic analyses, including WGCNA module construction, gene–trait correlation analysis, multi-database target intersection (GEO, GeneCards, TTD), molecular docking, and SPR analysis were used to identify candidate targets of Lico B. Subsequent in vitro and in vivo experiments—comprising Western blotting, qPCR, immunofluorescence staining, and flow cytometry in IL-17–stimulated keratinocytes and IMQ-induced psoriatic mice—were conducted to validate the therapeutic mechanisms of Lico B.

Article Snippet: The IL-17 group was treated with 100 ng/mL IL-17 (Proteintech; HZ-1113); the Lico B group received 9 μM Lico B (MCE; HY-N3388); the IL-17 + Lico B group was co-treated with IL-17 and Lico B; and the control group was treated with PBS.

Techniques: Biomarker Discovery, In Vitro, In Vivo, Western Blot, Immunofluorescence, Staining, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: Licoisoflavone B alleviates psoriasis via SCD1-targeted lipid metabolism reprogramming and suppression of Th17/IL-17–mediated inflammation

doi: 10.3389/fphar.2026.1754729

Figure Lengend Snippet: Schematic illustration of lico B treatment in psoriasis. Lico B modulates lipid metabolic pathways by suppressing SCD1-dependent metabolic reprogramming and reducing lipid droplet accumulation in keratinocyte, while simultaneously attenuating the TH17/IL-17 axis. Through concurrent regulation of keratinocyte metabolism and inflammatory cytokine production, Lico B ameliorate psoriatic skin pathology.

Article Snippet: The IL-17 group was treated with 100 ng/mL IL-17 (Proteintech; HZ-1113); the Lico B group received 9 μM Lico B (MCE; HY-N3388); the IL-17 + Lico B group was co-treated with IL-17 and Lico B; and the control group was treated with PBS.

Techniques: