Journal: Bioactive Materials

Article Title: Spatiotemporally programming the immune-osteogenic cascade with a dual-immunomodulatory scaffold for functional bone regeneration

doi: 10.1016/j.bioactmat.2026.04.002

Figure Lengend Snippet: Immune regulation and endogenous bone regeneration mechanism investigation. A) Network diagram showing the number of interactions between six subclusters. B) KEGG enrichment analysis of the upregulated DEGs in DIBS group compared to the HA group. C) Circular visualization of related pathway–gene enrichment analysis. D) Heatmap of key gene regulation in specific pathways. E) qRT-PCR validation for key gene expression in specific pathways. F) The interaction networks showing the correlation of representative immunomodulatory genes (CCL2, CCL20, Sfrp1, and Stat3, etc.) with angiogenesis/osteogenesis and macrophage regulation gene sets. G) Flow cytometry analysis and quantification of CCR2 F4/80 macrophage in peripheral blood. H) Immunofluorescence staining analysis of macrophage polarization inside scaffolds (one week after intramuscular implantation). I) Macrophage proliferation assay in a CCR2-dependent manner. J and K) Macrophage polarization assay in a CCR2-dependent manner. Data are represented as means ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; $ p < 0.05 (vs group without inhibitor), $$ p < 0.01 (vs group without inhibitor), $$$ p < 0.001 (vs group without inhibitor), $$$$ p < 0.0001 (vs group without inhibitor). ns, not significant.

Article Snippet: Throughout the experimental period, sustained CCR2 inhibition was achieved via daily intraperitoneal injections (2 mg/kg) of the highly selective CCR2 inhibitor RS504393 (Cat. No. HY-15418, MCE).

Techniques: Quantitative RT-PCR, Biomarker Discovery, Gene Expression, Flow Cytometry, Immunofluorescence, Staining, Proliferation Assay

Journal: Bioactive Materials

Article Title: Spatiotemporally programming the immune-osteogenic cascade with a dual-immunomodulatory scaffold for functional bone regeneration

doi: 10.1016/j.bioactmat.2026.04.002

Figure Lengend Snippet: Revascularization and osteogenesis are reinforced by M2 macrophage activation via the CCL2/CCR2 pathway. A and B) HUVECs and BMSCs proliferation assay under M2 macrophage activation. Created with BioRender.com . C) Migration assay and quantification of HUVECs. D) Tube formation assay and quantification of HUVECs. E and F) Early and later osteogenic differentiation of BMSC influenced by macrophage-induced microenvironment. Data are represented as means ± SD, ∗ p < 0.05 (vs Control), ∗∗ p < 0.01 (vs Control), ∗∗∗ p < 0.001 (vs Control), ∗∗∗∗ p < 0.0001 (vs Control); $ p < 0.05 (vs group without inhibitor), $$ p < 0.01 (vs group without inhibitor), $$$ p < 0.001 (vs group without inhibitor), $$$$ p < 0.0001 (vs group without inhibitor). ns, not significant.

Article Snippet: Throughout the experimental period, sustained CCR2 inhibition was achieved via daily intraperitoneal injections (2 mg/kg) of the highly selective CCR2 inhibitor RS504393 (Cat. No. HY-15418, MCE).

Techniques: Activation Assay, Proliferation Assay, Migration, Tube Formation Assay, Control

Journal: Oncology Letters

Article Title: A HRH1-YAP1 feedback loop drives pancreatic cancer progression and predicts therapeutic response

doi: 10.3892/ol.2026.15719

Figure Lengend Snippet: HRH1 is associated with prognosis, chemotherapy resistance and immunotherapy resistance in PDAC. (A) Venn analysis identified GPCR-related genes that are highly expressed and associated with prognosis in PDAC. (B) Kaplan-Meier analysis indicated the prognostic value of HRH1. (C) Online analysis using the GEPIA2 database showed HRH1 expression in pancreatic cancer and normal tissue (one-way ANOVA; *P<0.05). (D) HRH1 expression in tumor tissues and matched adjacent normal tissues. Data were obtained from 266 patients across seven cohorts (Wilcoxon rank-sum test; ***P<0.001). (E) Sequencing data from GSE26088 revealed that HRH1 expression was higher in 19 pancreatic cancer cell lines when compared with the normal pancreatic cell line (HPDE). (F) Reverse transcription quantitative-PCR analysis of HRH1 expression. Each of the 7 PDAC cell lines was compared with the control HPNE cell line. Statistical significance was assessed by one-way ANOVA followed by Dunnett's post hoc test (****P<0.0001; ***P<0.001). (G) Relationship between HRH1 expression and tumor mutation burden (Wilcoxon rank-sum test; **P<0.01). (H) In the CRA001160 dataset, HRH1 expression across different cell types, displayed using UMAP plot, feature plot and bar plot reflecting mean UMI count. (I) Relationship between HRH1 expression and TIDE score (Wilcoxon rank-sum test, **P<0.01). Association between HRH1 expression and resistance to (J) Gemcitabine, (K) Fluorouracil, (L) SN-38 and (M) Oxaliplatin. Statistical comparisons were performed using the Wilcoxon rank-sum test. GPCRs, G protein-coupled receptors; PDAC, pancreatic ductal adenocarcinoma; UMAP, Uniform Manifold Approximation and Projection.

Article Snippet: Fexofenadine (cat. no. HY-B0801; MedChemExpress) was employed as a selective HRH1 antagonist, while histamine (cat. no. HY-B1204; MedChemExpress) served as the HRH1 agonist.

Techniques: Expressing, Sequencing, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Mutagenesis

Journal: Oncology Letters

Article Title: A HRH1-YAP1 feedback loop drives pancreatic cancer progression and predicts therapeutic response

doi: 10.3892/ol.2026.15719

Figure Lengend Snippet: Targeting and inhibiting HRH1 is a potential therapeutic strategy for PDAC. (A) Scatter plot of the causal association between fexofenadine treatment and PDAC. (B) Leave-one-out sensitivity analysis using the IVW method. CCK-8 proliferation assay was used to detect the proliferation of (C) PANC1 cells and (D) SW1990 cells after HRH1 knockdown. The knockdown groups were compared with the non-targeting control in PANC-1 and SW1990 cells. Statistical significance was assessed by one-way ANOVA followed by Dunnett's post hoc test (****P<0.0001). Experiments were performed in triplicate. The reverse transcription-quantitative PCR validation data for HRH1 knockdown efficiency is provided in . Dose-response curves of (E) PANC1 and (F) SW1990 cells transfected with siControl, siHRH1#1 or siHRH1#2 and treated with increasing concentrations of gemcitabine for 48 h. Cell viability was assessed by CCK-8 assay and normalized to the untreated control. PDAC, pancreatic ductal adenocarcinoma; CCK-8, Cell Counting Kit-8; si, small interfering; IVW, inverse variance-weighted.

Article Snippet: Fexofenadine (cat. no. HY-B0801; MedChemExpress) was employed as a selective HRH1 antagonist, while histamine (cat. no. HY-B1204; MedChemExpress) served as the HRH1 agonist.

Techniques: CCK-8 Assay, Proliferation Assay, Knockdown, Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Biomarker Discovery, Transfection, Cell Counting

Journal: Oncology Letters

Article Title: A HRH1-YAP1 feedback loop drives pancreatic cancer progression and predicts therapeutic response

doi: 10.3892/ol.2026.15719

Figure Lengend Snippet: HRH1 is associated with YAP1 signaling. (A) Metascape enrichment analysis revealed that HRH1 is functionally associated with YAP1 signaling. The gene set was derived from differentially upregulated genes in the HRH1-high expression group within TCGA-PAAD cohort. Gene Set Enrichment Analysis further indicated a significant correlation between HRH1 and both the (B) ‘Cordenosi YAP conserved signature’ and the (D) ‘YAP1 up signature’. (C) A heatmap illustrated HRH1 expression and the expression of multiple YAP1 pathway target genes. Based on HRH1 expression levels, TCGA PDAC groups were categorized into two groups to compare the expression of YAP1-targeted genes (***P<0.001). Expression changes of HRH1, CTGF, CYR61, ANKRD1 and YAP1 were assessed following knockdown of HRH1 and YAP1. The knockdown groups were compared with the non-targeting control in (E) PANC-1 and (F) SW1990 cells. Statistical significance was assessed by one-way ANOVA followed by Dunnett's post hoc test ( ***P <0.001, **P<0.01; ns, Not Significant). si, small interfering; NC, negative control.

Article Snippet: Fexofenadine (cat. no. HY-B0801; MedChemExpress) was employed as a selective HRH1 antagonist, while histamine (cat. no. HY-B1204; MedChemExpress) served as the HRH1 agonist.

Techniques: Derivative Assay, Expressing, Knockdown, Control, Negative Control

Journal: Oncology Letters

Article Title: A HRH1-YAP1 feedback loop drives pancreatic cancer progression and predicts therapeutic response

doi: 10.3892/ol.2026.15719

Figure Lengend Snippet: The reciprocal regulation between YAP1 and HRH1. (A) YAP1 binds to the promoter region of HRH1. The ChIP-seq data for H3K4me3, H3K27ac and YAP1, as well as ATAC-seq data, were obtained from the ChIP-Atlas database. (B) In PA-TU-8902 and PCa3 cells, knockdown of YAP1 and TAZ resulted in reduced chromatin accessibility at the HRH1 promoter region. ATAC-seq data for PA-TU-8902 and PCa3 were obtained from the ChIP-Atlas database. Knockdown of HRH1 and YAP1 led to a marked reduction in YAP1 protein levels in (C) SW199 and (D) PANC10 cells. The observed molecular weights of β-actin and YAP1 were ~42 and 75 kDa, respectively. Relative YAP1 expression levels, normalized to β-actin and the NC, are provided in . Treatment of (E) SW1990 and (F) PANC1 cells with fexofenadine and histamine resulted in altered expression of YAP1. Relative YAP1 expression levels, normalized to β-actin and the Control group, are provided in . si, small interfering; NC, negative control.

Article Snippet: Fexofenadine (cat. no. HY-B0801; MedChemExpress) was employed as a selective HRH1 antagonist, while histamine (cat. no. HY-B1204; MedChemExpress) served as the HRH1 agonist.

Techniques: ChIP-sequencing, Knockdown, Expressing, Control, Negative Control

Journal: Oncology Letters

Article Title: A HRH1-YAP1 feedback loop drives pancreatic cancer progression and predicts therapeutic response

doi: 10.3892/ol.2026.15719

Figure Lengend Snippet: Prognostic value of the HRH1/YAP1 signaling axis-derived signature in TCGA, ICGC and GEO cohorts. (A) Risk score distribution plots demonstrating the correlation between elevated risk scores and mortality events. (B) Kaplan-Meier curves stratified by risk groups. (C) Time-dependent receiver operating characteristic curves assessing the model's predictive accuracy for 1-, 2-, 3- and 4-year survival. (D) Expression heatmaps of the eight signature genes incorporated in the risk model. (E) PCA visualizing the separation between high- and low-risk groups. PCA, principal component analysis; KM, Kaplan-Meier. TCGA, The Cancer Genome Atlas; ICGC, International Cancer Genome Consortium; GEO, Gene Expression Omnibus.

Article Snippet: Fexofenadine (cat. no. HY-B0801; MedChemExpress) was employed as a selective HRH1 antagonist, while histamine (cat. no. HY-B1204; MedChemExpress) served as the HRH1 agonist.

Techniques: Derivative Assay, Expressing, Gene Expression