Fig 2: The dynamic of histopathological changes and liver regeneration in a single infection mouse model with S. japonicum. A Representative images of HE staining at different infection time points. Arrows indicate the necrosis. B The statistical analysis of percent of granulomas area. n = 7 in uninfected group, n = 5 in each infected group. C Representative images of Masson staining. D The statistical analysis of percents of fibrosis area. n = 7 in uninfected group, n = 5 in each infected group. E Representative images of TUNEL staining. n = 3 mice/group. F Representative images of CD45 staining. n = 3 mice/group. G Representative images of F4/80 staining. n = 3 mice/group. H The expression level of albumin in serum. n = 7 in uninfected group, n = 5 mice in each infected group. Images were captured by an inverted microscope (Olympus, BX63, Tokyo, Japan). *P < 0.05, **P < 0.01, ***P < 0.001

Fig 3: The hydrogel microsphere vaccine amplified CD8+ T-cell-specific antitumour immunity.a Representative images showing the t-SNE analysis of CD8, CD4, IFN-?, TNF-a and Ki67 markers in TdLNs across different treatment groups. b Identification of the proportion of CD8+ T cells (left), Ki67+CD8+ T cells (middle) and IFN-?+CD8+ T cells in the TdLNs. c Representative images showing the IF multicolour-labelled CD8-, CD4- and CK19-positive cells in pancreatic tumour tissue in tumour-bearing mice treated with the hydrogel microsphere vaccine or control hydrogel microspheres. Five biologically independent samples were performed to confirm data stability. d Representative images of flow cytometry analysis showing the proportion of tumour-infiltrating CD8+ T cells and CD4+ T cells across different treatment groups. e Representative images showing the t-SNE analysis of the distribution of tumour-infiltrating NK cells (NK-1.1+CD45+, purple), total myeloid cells (CD11c+Ly6G+ CD45+, green), CD8+ T cells (CD8+CD3+CD45+, red) and CD4+ T cells (CD4+CD3+CD45+, blue) across different treatment groups. (f) Identification of the cellular density of tumour-infiltrating CD8+ T cells (top left), FoxP3+CD25+ Tregs (top right), IFN-?+CD8+ T cells (middle right), Ki67+CD8+ T cells (bottom left) and Gzmb+CD8+ T cells (bottom right) across different treatment groups. g–i Quantification of intratumour and serum levels of IL12p70 (g), IFN-? (h), and TNF-a (i) level. j Identification of serum ALT and AST values. b, f, g–i n = 5 biological independent samples for each group. Data are presented as mean values +/- SEM. Two-tailed Dunnett t test. j n = 5 biological independent samples for each group. Data are presented as mean values +/- SEM. Two-tailed unpaired t test. Source data are provided as a Source data file.

Fig 4: Preparation and functional validation of the hydrogel microsphere vaccine in vitro.a Schematic diagram of the preparation of Cas9@liposome nanoparticles. b Representative TEM images of the plasmid@lip nanoparticles. c, d Encapsulation efficiency (c), particle size distribution and PDI value (d) of the Cas9@lip nanoparticles. n = 3 independent experiments. Data are presented as mean values +/- SD. e Schematic diagram of the preparation of the CD40L@CaCO3 nanoparticles. f Representative scanning electron microscope (SEM) images of the CD40L@CaCO3 nanoparticles. g Identification of the hydrodynamic diameter of the CD40L@CaCO3 nanoparticles using the Malvern–Zetasizer method. h Identification of the hydrodynamic diameter of the HOOC–PLGA–PEG–PLGA–COOH solution. i Schematic diagram of the preparation of the hydrogel microsphere vaccine. j Representative scanning electron microscope (SEM) images of the hydrogel microspheres. k Enlarged representative SEM image (left panel) and regional element analysis (right panel) of the hydrogel microsphere vaccine. l The release of FLT3L and CD40L by the hydrogel microsphere vaccine in vitro (pH = 6.8). n = 3 independent experiments. Data are presented as mean values +/- SD. m Schematic diagram of cell transfection analysis using the combination of the hydrogel microsphere vaccine and electroporation. n Representative images of flow cytometry experiments identifying GFP-positive KPC cells in different treatment groups (o). o Analysis of the transfection efficiency of 3T3\KPC\AsPC1\Panc02 cells in different treatment groups. n = 3 biologically independent samples. p Schematic diagram illustrating the effect of hydrogel microsphere vaccines and different hydrogel counterparts on bone marrow-derived dendritic cells (BMDCs) in vitro. q Analysis of the effect of hydrogel microsphere vaccines and hydrogel microsphere counterparts on the expression of MHC-I, CD40, CD80 and CD86 on the surface of total DCs (CD24+CD64-CD11c+MHCII+CD45+alive) and cDC1s (CD103+CD11b-CD24+CD64-CD11c+MHCII+CD45+alive) in different treatment groups. n = 4 biologically independent samples. b, f, j, k Three independent experiments were performed to confirm data stability. o, q Data are presented as mean values +/- SEM. Two-tailed unpaired t test. Source data are provided as a Source data file.

Fig 5: Metabolic reprogramming from oxidative phosphorylation (OXPHOS) to glycolysis had been found during glandular aging, which was associated with senescence-associated secretory phenotype (SASP) and immune response(A) The bubble chart of the results of gene set enrichment analysis (GSEA) was generated using R package ‘ggplot2’. The top 13 enriched KEGG pathways in response to phenotype aged of RNA-seq data were listed, in the order of normalized enriched score (NES).(B) The top ranked gene set in the standard GSEA analysis was glycolysis glyconeogenesis gene set. Enrichment score was shown on the y axis as the relevant genomic features traversed on the x axis. FDR, false discovery rate.(C) Co-localization by confocal microscopy of mitochondrial protein COXIV (green), NKCC1 (red), and DAPI (blue) in young and aged minor salivary glands. Scale bars: 25 µm, n = 8 per group.(D) Co-localization by confocal microscopy of COXIV (green), CK7 (red), and DAPI (blue) in young and aged minor salivary glands. Scale bars: 25 µm, n = 8 per group.(E) Correlation of OXPHOS genes, cell cycling & DNA replication genes, glycolysis and fatty acid metabolism genes with SASP factors and salivary gland function genes. Correlation statistics and the related p value were computed using Spearman correlation. *p < 0.05, **p < 0.01.(F) IHC staining for IL-6. Scale bars: 100 µm, n = 12 per group.(G) IHC staining for TNF-a. Scale bars: 100 µm, n = 12 per group.(H) Western blot for IL-6 and TNF-a between young and aged groups. Bar graph showed semi-quantitative analysis of western blot about IL-6 and TNF-a protein expression. Relative protein expression is represented as the optical density ratio of targeted protein to actin. Data are expressed as the mean ± SD. ***p < 0.001, vs. young group (n = 3). All experiments had been repeated at least twice, with similar results.(I) MMP9/CD45 co-localization was visualized by immunofluorescence (IF) staining and IHC for CD3, CD8, cytochrome c, and Bcl-2. Scale bars: 25 µm (IF); 100 µm (IHC), n = 12 per group; (J) Semi-quantitative IF or IHC analysis for showing percentage of MMP9, CD45, CD3, and CD8 positive cells using ImageJ in young and aged labial salivary glands. Error bars indicate SD. *p < 0.05, **p < 0.01, ***p < 0.01, n = 12.