Journal: Journal of Advanced Research

Article Title: Gut microbiota-derived xanthohumol protects against heatstroke by inhibiting macrophage pyroptosis in mice

doi: 10.1016/j.jare.2025.07.031

Figure Lengend Snippet: Dysbiosis of the gut microbiota contributes to heatstroke-induced organ injury in mice. (A) PCoA plot based on the weighted UniFrac distance matrices in mice treated with or without heatstroke. n = 8. (B–C) Relative abundance of bacteria at the phylum level (B) and the genus level (C) in the cecal contents of mice. n = 8. (D) LEfSe analysis between the two groups. n = 8. (E) Schematic process of FMT experiment. Tissue samples were harvested 12 h later. (F) Plasma concentration of ALT, AST, and Cr, and total protein contents in BALF. Experimental design as in (1E). n = 5. (G) H&E staining of the liver, kidney, and lung. The box plots showed the quantitative analysis histopathological score in each group. Scale bars, 100 μm. n = 6. (H) TUNEL staining and quantification of dead cells of the liver, kidney, and lung. Experimental design as in (1E). Scale bars, 100 μm. n = 6. (I) Plasma levels of TNF-α, IL-1β, and IL-6 in each group. Experimental design as in (1E). n = 5–6. (J) Immunohistological staining of and quantification of CD68 + cells in the lung tissue. Experimental design as in (1E). Scale bars, 100 μm. n = 5–6. Data are represented as the mean ± SEM. * p < 0.05 were determined by adonis analysis and anosim analysis in A, two-tailed Student's t -test in F–J.

Article Snippet: Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was measured by a commercial kit (Beyotime, China) as previously described [ ].

Techniques: Bacteria, Clinical Proteomics, Concentration Assay, Staining, TUNEL Assay, Two Tailed Test

Journal: Journal of Advanced Research

Article Title: Gut microbiota-derived xanthohumol protects against heatstroke by inhibiting macrophage pyroptosis in mice

doi: 10.1016/j.jare.2025.07.031

Figure Lengend Snippet: Decreased abundances of L. murinus and XN are induced by heatstroke. (A and B) PCA scatter plots and volcano plots of non-targeted metabolomics analysis in the cecal contents of mice treated with or without heatstroke exposure. n = 6. (C) Molecular structure of XN. (D–F) Measurements of XN in the cecum and plasma of mice treated with or without heatstroke exposure were determined by LC-MS/MS. n = 6. (G) The concentration of XN in the cecum of mice treated with or without ABX was determined by LC-MS/MS. n = 4. (H) The relative abundances of BA, AKK, Lachnospiraceae bacterium_28_4 , and L. murinus in control and heatstroke-treated mice were determined by RT-PCR. n = 6. (I–K) Measurement of XN in the culture supernatants with the mouse chow (MC) was determined by LC-MS/MS. n = 5. (L) Measurement of XN in the feces of mice treated with or without L. murinus . n = 6. (M) Schematic process of the L. murinus intervention: heatstroke-treated mice were pretreated with daily oral administration of L. murinus for 7 days. Tissue samples were harvested 12 h later. (N) Plasma ALT, AST, and Cr levels, and total protein contents in BALF. Experimental design as in (2 M). n = 6. (O) H&E staining and histopathological score of the liver, kidney, and lung. Experimental design as in (2 M). Scale bars, 100 μm. n = 6. (P) TUNEL staining and quantification of dead cells of the liver, kidney, and lung. Experimental design as in (2 M). Scale bars, 100 μm. n = 6. Data are represented as the mean ± SEM. * p < 0.05, ns p > 0.05 were determined by adonis analysis and anosim analysis in A, two-tailed Student's t -test in E–L, N–P.

Article Snippet: Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was measured by a commercial kit (Beyotime, China) as previously described [ ].

Techniques: Clinical Proteomics, Liquid Chromatography with Mass Spectroscopy, Concentration Assay, Control, Reverse Transcription Polymerase Chain Reaction, Staining, TUNEL Assay, Two Tailed Test

Journal: Journal of Advanced Research

Article Title: Gut microbiota-derived xanthohumol protects against heatstroke by inhibiting macrophage pyroptosis in mice

doi: 10.1016/j.jare.2025.07.031

Figure Lengend Snippet: Gut microbiota-derived XN protects against heatstroke in a murine model. (A) Schematic process of the XN administration: heatstroke-treated mice were pretreated with XN orally once a day for 3 days. Tissue samples were harvested 12 h later. (B) Plasma ALT, AST, and Cr levels, and MPO activity in the lung. Experimental design as in (3A). n = 7. (C) H&E staining and histopathological scores of the liver, kidney, and lung. Experimental design as in (3A). Scale bars, 100 μm. n = 7. (D) TUNEL staining and quantification of dead cells of the liver, kidney, and lung. Experimental design as in (3A). Scale bars, 100 μm. n = 7. (E) The serum levels of TNF-α, IL-1β, and IL-6 in each group. Experimental design as in (3A). n = 6. (F) Immunohistological staining of and quantification of CD68 + cells in the lung tissue. Experimental design as in (3A). Scale bars, 100 μm. n = 6. Data are represented as the mean ± SEM. * p < 0.05 were determined by one-way ANOVA (Tukey's test) in B–F.

Article Snippet: Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was measured by a commercial kit (Beyotime, China) as previously described [ ].

Techniques: Derivative Assay, Clinical Proteomics, Activity Assay, Staining, TUNEL Assay

Journal: Journal of Advanced Research

Article Title: Gut microbiota-derived xanthohumol protects against heatstroke by inhibiting macrophage pyroptosis in mice

doi: 10.1016/j.jare.2025.07.031

Figure Lengend Snippet: XN treatment suppresses macrophage pyroptosis in heatstroke. (A) Schematic process of the experimental approach: heatstroke-treated mice were pretreated with oral XN once a day for 3 days, and mice were treated with 200 mL LIPO intraperitoneally 24 h before heatstroke exposure. Tissue samples were harvested 12 h later. (B) Plasma ALT, AST, and Cr levels, and total protein contents in BALF. Experimental design as in (4A). n = 6. (C) H&E staining and histopathological score of the liver, kidney, and lung. Experimental design as in (4A). Scale bars, 100 μm. n = 6. (D) TUNEL staining and quantification of dead cells of the liver, kidney, and lung. Experimental design as in (4A). Scale bars, 100 μm. n = 6. (E) Representative TEM images of peritoneal macrophages in mice. Red arrows: the peritoneal macrophage pyroptosis. Experimental design as in (4A). Scale bars, 2 μm. (F) The mRNA levels of Nlrp3, Casp1, Gsdmd, Nlrp6, and Txnip in sorting PMs (peritoneal macrophages) after heatstroke exposure. n = 5. Data are represented as the mean ± SEM. * p < 0.05, ns p > 0.05 were determined by one-way ANOVA (Tukey's test) in B–D, unpaired two-tailed Student's t -test in F. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was measured by a commercial kit (Beyotime, China) as previously described [ ].

Techniques: Clinical Proteomics, Staining, TUNEL Assay, Two Tailed Test

Journal: PLOS One

Article Title: Impact of blast exposure on visual pathway: Mechanism exploration and novel diagnostic perspectives

doi: 10.1371/journal.pone.0344993

Figure Lengend Snippet: (A) Schematic diagram of the visual pathway and key observation areas in its intracranial segment. (B) Representative images of immunofluorescence staining for NeuN (green) and NLRP3 (red), Iba-1 (grey) in mouse visual cortex slices, Group information is shown in images. The red triangle marks indicate the co-localization of NLRP3 and NeuN. The scale bars for all fluorescence intensity channels and the merge are set at 50 μm, the magnified inset is 20μm. Quantitative analysis of visual cortex immunofluorescence density in Iba-1 (C) and NLRP3 (G) . (H) Fluorescence intensity of NeuN + NLRP3 + / NeuN + , n = 4. (D, E, F, I, J) Western blot analysis of Iba-1, IL-1β, cleaved Gasdermin D and NLRP3 in visual cortex lysates, the molecular mass is indicated in kilodaltons, n = 3. (K-L) Quantitative analysis of representative fluorescence images of NeuN staining with TUNEL labeling in the peri-contusion region from different groups that information is shown in images, Scale bar = 100 μm, n = 4. (M) Quantitative analysis of visual cortex NeuN + cells, n = 4. Error bars indicate mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, ns p > 0.05.

Article Snippet: Brain sections were subjected to double staining using NeuN antibody and the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay kit (Beyotime, #C1090) to identify DNA damage, following the manufacturer’s protocol.

Techniques: Immunofluorescence, Staining, Fluorescence, Western Blot, TUNEL Assay, Labeling

Journal: PLOS One

Article Title: Impact of blast exposure on visual pathway: Mechanism exploration and novel diagnostic perspectives

doi: 10.1371/journal.pone.0344993

Figure Lengend Snippet: (A) Representative images of immunofluorescence staining for NeuN (green), NLRP3 (red), and Iba-1 (grey) in mouse visual cortex slices, as well as group information, are shown in the images. The red triangle marks indicate the co-localization of NLRP3 and NeuN. The scale bars for all fluorescence intensity channels and the merge are set at 50 μm, the magnified inset is 20 μm, n = 4. (B) Quantitative analysis of visual cortex immunofluorescence density in NLRP3, n = 4. (C) Fluorescence intensity of NeuN + NLRP3 + /NeuN + , n = 4. (D) Quantitative analysis of visual cortex immunofluorescence density in Iba-1, n = 4. (E, F, G, H) Western blot analysis of Iba-1, cleaved Gasdermin D and NLRP3 in visual cortex lysates. The molecular mass is indicated in kilodaltons, n = 3. (I, J) Quantitative analysis of representative fluorescence images of NeuN staining with TUNEL labeling in the peri-contusion region from different groups that information is shown in images, Scale bar = 100 μm, n = 4. (K) Quantitative analysis of visual cortex NeuN+ cells. (L) Representative images of immunofluorescence staining for MBP (green) and βIII-Tubulin (red) in mouse optic nerve slices. Group information is shown in images. Scale bar = 20 μm, n = 4. (M) optic nerve MBP density in different groups, n = 4. (N) optic nerve relative fluorescence intensity of MBP + /β III Tubulin + in different groups, n = 4. (O) TEM analysis of the optic nerve between the BE 24 h and BE 28 d groups, red stars indicate axons with obvious demyelination. Error bars indicate mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, ns p > 0.05.

Article Snippet: Brain sections were subjected to double staining using NeuN antibody and the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay kit (Beyotime, #C1090) to identify DNA damage, following the manufacturer’s protocol.

Techniques: Immunofluorescence, Staining, Fluorescence, Western Blot, TUNEL Assay, Labeling