Journal: Clinical and Translational Medicine

Article Title: Macrophage‐derived galectin‐3 contributes to pyroptosis, apoptosis and necroptosis through TLR4/MyD88/NF‐κB/NLRP3 during atherosclerosis

doi: 10.1002/ctm2.70637

Figure Lengend Snippet: Galectin‐3 expression is abundant in human and mouse atherosclerotic lesions. (A) Ten main cell types are visualised in atherosclerotic core (AC) and proximal adjacent (PA) tissues by tSNE (t‐distributed stochastic neighbour embedding). (B) The macrophage population significantly increased in AC relative to PA. (C) Biaxial scatter plots show the expression pattern of galectin‐3 in total cell types between AC and PA. The colour scale represents expression levels in biaxial scatter plots (grey: low; pink: high). (D) Galectin‐3‐positive macrophages expanded in AC in comparison with PA. (E) Five macrophage subtypes are visualised in AC and PA tissues by tSNE. (F) My.0 and My.1 account for 34.1% and 47.6% of macrophages in AC, respectively. My.2 significantly increased in AC relative to PA. (G) Biaxial scatter plots exhibit the expression pattern of galectin‐3 in macrophage subtypes between AC and PA. (H) Galectin‐3‐positive My.0 and My.1 account for 35.8% and 47.5% of galectin‐3‐positive macrophages in AC, respectively. Galectin‐3‐positive My.2 expands in AC in comparison with PA. (I) Representative Western blots and relative quantitative analysis of galectin‐3 in human atherosclerotic lesions and peripheral normal artery. (J) Triple immunofluorescence staining for galectin‐3 (red), NLRP3 (green), CD68 (pink) and DAPI (blue) in human atherosclerosis and peripheral normal artery reveals the colocalisation of galectin‐3 and NLRP3 in CD68‐positive macrophages. Scale bar: 50 µm. (K) Representative Western blots and relative quantitative analysis of galectin‐3 in the aortas of ApoE −/− mice fed with an HFD or normal diet. (L) Triple immunofluorescence staining for galectin‐3 (red), NLRP3 (green), CD68 (pink) and DAPI (blue) in human atherosclerosis and peripheral normal artery reveals that galectin‐3 and NLRP3 are colocalised in CD68‐positive macrophages. Scale bar: 50 µm. (M) Cell lysates from ox‐LDL‐treated macrophages are immuno‐precipitated with anti‐galectin‐3 or anti‐NLRP3 antibodies, and blotted with anti‐NLRP3 or anti‐galectin‐3 antibodies. Data are derived from three to five independent experiments. * p ˂.05, ** p ˂.01, *** p ˂.001 by Student's t test. ns: not significant.

Article Snippet: Galectin‐3 and NLRP3 expression levels are elevated in both human and murine atherosclerotic lesions.

Techniques: Expressing, Comparison, Western Blot, Immunofluorescence, Staining, Derivative Assay

Journal: Clinical and Translational Medicine

Article Title: Macrophage‐derived galectin‐3 contributes to pyroptosis, apoptosis and necroptosis through TLR4/MyD88/NF‐κB/NLRP3 during atherosclerosis

doi: 10.1002/ctm2.70637

Figure Lengend Snippet: Pyroptosis, apoptosis and necroptosis in macrophages concurrently exist in human atherosclerotic lesions. (A–C) Necrotic core formation, lipid deposition and extracellular fibrosis are observed in lower extremity and carotid atherosclerotic lesions, but not in histologically normal artery by means of HE, Oil Red O, and Movat's staining. Scale bar: 1000 µm. (D) Immunohistochemical staining for CD68, a marker for macrophages, is conducted on histologically normal arteries as well as on lower extremity and carotid atherosclerotic lesions. Scale bar: 100 µm. (E–G) Compared with histologically normal artery (E), lower extremity (F) and carotid atherosclerotic lesions (G) show the pyroptotic, apoptotic and necroptotic characteristics of macrophages, including discontinuity of plasma membrane (red arrows), chromatin condensation, and electron‐light zones in transmission electron microscopy images. Scale bar: 2.5 µm. (H) Representative Western blots and relative quantitative analysis of NLRP3, GSDMD and GSDMD‐N in human atherosclerotic lesions and peripheral normal artery. (I) Representative Western blots and relative quantitative analysis of caspase‐3, cleaved caspase‐3, caspase‐8 and cleaved caspase‐8 in human atherosclerotic lesions and peripheral normal artery. (J) The capacity of caspase‐3 in human atherosclerotic lesions and peripheral normal artery. (K and L) Representative Western blots and relative quantitative analysis of RIPK3, MLKL and phospho‐MLKL in human atherosclerotic lesions and peripheral normal artery. (M) Representative Western blots and relative quantitative analysis of TLR4, MyD88 and phospho‐NF‐kB in human atherosclerotic lesions and peripheral normal artery. (N) Human atherosclerotic lesions release a significant amount of TNF‐1α, IL‐1β, IL‐18 and IL‐6, whereas the peripheral normal artery releases less. (O) Triple immunofluorescence staining for GSDMD (green), caspase‐3 (red), RIPK3 (yellow) and DAPI (blue) in human atherosclerosis and peripheral normal artery reveals the co‐existence of pyroptosis, apoptosis and necroptosis as evidenced by the colocalisation of GSDMD, caspase‐3 and RIPK3. Triple‐positive cells are shown by the arrows. Scale bar: 50 µm. (P–R) Double immunofluorescence staining for GSDMD (M)/caspase‐3 (N)/RIPK3 (O) (red), CD68 (green), and DAPI (blue) in human atherosclerosis and peripheral normal artery demonstrates the presence of pyroptotic, apoptotic and necroptotic markers in macrophages, as indicated by the colocalisation of caspase‐3/GSDMD/RIPK3 and CD68 (a macrophage marker). Double‐positive cells are shown by the arrows. Scale bar: 50 µm. Data are derived from three to five independent experiments. * p ˂.05, ** p ˂.01, *** p ˂.001 by Student's t test. ns: not significant.

Article Snippet: Galectin‐3 and NLRP3 expression levels are elevated in both human and murine atherosclerotic lesions.

Techniques: Staining, Immunohistochemical staining, Marker, Clinical Proteomics, Membrane, Transmission Assay, Electron Microscopy, Western Blot, Immunofluorescence, Double Immunofluorescence Staining, Derivative Assay

Journal: Clinical and Translational Medicine

Article Title: Macrophage‐derived galectin‐3 contributes to pyroptosis, apoptosis and necroptosis through TLR4/MyD88/NF‐κB/NLRP3 during atherosclerosis

doi: 10.1002/ctm2.70637

Figure Lengend Snippet: Silencing galectin‐3 downregulated TLR4/MyD88/NF‐kB expression and attenuated ox‐LDL induced pyroptotic, apoptotic, and necroptotic cell death in macrophages. (A) Electron microscopy ultrastructural analysis of control and ox‐LDL‐induced macrophages. Control macrophages have a normal‐looking cellular structure, whereas ox‐LDL‐induced macrophages show loss of cell plasma integrity, chromatin condensation or fragmentation, and electron‐light zone. Scale bar: 2.5 µm. (B) Confocal microscopy with double immunofluorescence staining for caspase‐3 (red) and RIPK3 (green) in macrophages show the colocalisation of apoptotic and necroptotic components. Confocal microscopy analysis of double immunofluorescence labelling is indicative of overlapping expression of caspase‐3 (red) and GSDMD (green) in macrophages. Scale bar: 25 µm. (C) Representative Western blots and relative quantitative analysis of galectin‐3 in control macrophages and cells treated with ox‐LDL, ox‐LDL plus siControl RNA, and ox‐LDL plus siGalectin‐3 RNA. (D and E) Flow cytometry (E) and quantification analysis (F) with annexin V/PI double staining show that ox‐LDL increased the percentage of apoptotic cells in macrophages, which is alleviated by silencing galectin‐3. (F–H) Flow cytometry (F) and quantification analysis (G) with PI/Hoechst staining (H) show that ox‐LDL enhanced PI uptake in macrophages, which is markedly blocked by silencing galectin‐3. Scale bar: 50 µm. (I) Silencing galectin‐3 abrogated LDH release in macrophages ignited by ox‐LDL. (J) Ox‐LDL induced the accumulation of intracellular lipid droplets in macrophages, which are potently reversed by silencing galectin‐3. Scale bar: 50 µm. (K) Representative Western blots and relative quantitative analysis of NLRP3, GSDMD and GSDMD‐N in control macrophages and cells treated with ox‐LDL, ox‐LDL plus siControl RNA, and ox‐LDL plus siGalectin‐3 RNA. (L) Representative Western blots and relative quantitative analysis of caspase‐3, cleaved caspase‐3, caspase‐8 and cleaved caspase‐8 in control macrophages and cells treated with ox‐LDL, ox‐LDL plus siControl RNA, and ox‐LDL plus siGalectin‐3 RNA. (M and N) Representative Western blots and relative quantitative analysis of RIPK3, MLKL and phospho‐MLKL in control macrophages and cells treated with ox‐LDL, ox‐LDL plus siControl RNA, and ox‐LDL plus siGalectin‐3 RNA. (O) Ox‐LDL treatment promotes the release of proinflammatory cytokines (TNF‐1α, IL‐1β, IL‐18 and IL‐6) from macrophages, which is markedly rescued by silencing galectin‐3. (P and Q) Representative Western blots and relative quantitative analysis of TLR4, MyD88, NF‐kB and phospho‐NF‐kB in control macrophages and cells treated with ox‐LDL, ox‐LDL plus siControl RNA, and ox‐LDL plus siGalectin‐3 RNA. (R) Cell lysates from ox‐LDL‐treated macrophages are immunoprecipitated with anti‐TLR4 or anti‐MyD88 antibodies, and blotted with anti‐TLR4 or anti‐MyD88 antibodies. Data are derived from three to five independent experiments. * p ˂.05, ** p ˂.01, *** p ˂.001 by Student's t test. ns: not significant.

Article Snippet: Galectin‐3 and NLRP3 expression levels are elevated in both human and murine atherosclerotic lesions.

Techniques: Expressing, Electron Microscopy, Control, Clinical Proteomics, Confocal Microscopy, Double Immunofluorescence Staining, Immunofluorescence, Western Blot, Flow Cytometry, Double Staining, Staining, Immunoprecipitation, Derivative Assay

Journal: Clinical and Translational Medicine

Article Title: Macrophage‐derived galectin‐3 contributes to pyroptosis, apoptosis and necroptosis through TLR4/MyD88/NF‐κB/NLRP3 during atherosclerosis

doi: 10.1002/ctm2.70637

Figure Lengend Snippet: NLRP3 agonist nigericin counteracted the inhibitory effect of silencing galectin‐3 on pyroptosis, apoptosis and necroptosis in macrophages. (A) Confocal microscopy with double immunofluorescence staining for galectin‐3 (red) and NLRP3 (green) in macrophages reveals the colocalisation of galectin‐3 with NLRP3. Scale bar: 25 µm. (B and C) Flow cytometry (B) and quantification analysis (C) with annexin V/PI double staining show that silencing galectin‐3 decreases the percentage of apoptotic cells in ox‐LDL‐induced macrophages, and nigericin robustly blunts the inhibitory effect of siGalectin‐3. (D–F) Flow cytometry (D) and quantification analysis (E) with PI/Hoechst staining (F) show that silencing galectin‐3 diminishes the percentage of PI‐positive cells in ox‐LDL‐induced macrophages, and nigericin mostly abolishes the protective effect of siGalectin‐3. Scale bar: 50 µm. (G) Silencing galectin‐3 suppresses the LDH release in ox‐LDL‐induced macrophages, which is largely abrogated by nigericin. (H) Silencing galectin‐3 lessens the intracellular lipid droplet in ox‐LDL‐induced macrophages, while nigericin exerts the opposite effect. Scale bar: 50 µm. (I) Representative Western blots and relative quantitative analysis of NLRP3, GSDMD and GSDMD‐N in macrophages treated with ox‐LDL, ox‐LDL plus galectin‐3 siRNA, ox‐LDL plus nigericin, and ox‐LDL plus galectin‐3 siRNA plus nigericin. (J) Representative Western blots and relative quantitative analysis of caspase‐3, cleaved caspase‐3, caspase‐8 and cleaved caspase‐8 in macrophages treated with ox‐LDL, ox‐LDL plus galectin‐3 siRNA, ox‐LDL plus nigericin, and ox‐LDL plus galectin‐3 siRNA plus nigericin. (K) The activity of caspase‐3 in macrophages treated with ox‐LDL, ox‐LDL plus galectin‐3 siRNA, ox‐LDL plus nigericin, and ox‐LDL plus galectin‐3 siRNA plus nigericin. (L and M) Representative Western blots and relative quantitative analysis of RIPK3, MLKL and phospho‐MLKL in macrophages treated with ox‐LDL, ox‐LDL plus galectin‐3 siRNA, ox‐LDL plus nigericin, and ox‐LDL plus galectin‐3 siRNA plus nigericin. (N) Silencing galectin‐3 inhibits the release of inflammatory cytokines (TNF‐1α, IL‐1β, IL‐18 and IL‐6) in ox‐LDL‐induced macrophages, and nigericin effectively blocks the role of siGalectin‐3. Data are derived from three to five independent experiments. * p ˂.05, ** p ˂.01, *** p ˂.001 by Student's t test. ns: not significant.

Article Snippet: Galectin‐3 and NLRP3 expression levels are elevated in both human and murine atherosclerotic lesions.

Techniques: Confocal Microscopy, Double Immunofluorescence Staining, Flow Cytometry, Double Staining, Staining, Western Blot, Activity Assay, Derivative Assay

Journal: Clinical and Translational Medicine

Article Title: Macrophage‐derived galectin‐3 contributes to pyroptosis, apoptosis and necroptosis through TLR4/MyD88/NF‐κB/NLRP3 during atherosclerosis

doi: 10.1002/ctm2.70637

Figure Lengend Snippet: Pyroptosis, apoptosis and necroptosis in macrophages coordinately occurred in ApoE −/− mice fed an HFD, which are alleviated by galectin‐3 deficiency, and conversely are aggravated by NLRP3 agonist nigericin. (A) Pyroptosis, apoptosis and necroptosis of macrophages are identified in the aortas of ApoE −/− mice fed an HFD, as evidenced by plasma membrane pore (red arrows), chromatin condensation (red arrows), and electron‐light zone (red arrows) by transmission electron microscopy. Scale bar: 2.5 µm. (B) Triple immunofluorescence staining for GSDMD (green), caspase‐3 (red), RIPK3 (pink) and DAPI (blue) in the aortas of ApoE −/− mice fed HFD or normal diet reveals the potential crosstalk among pyroptosis, apoptosis and necroptosis as evidenced by the colocalisation of GSDMD, caspase‐3 and RIPK3. Three‐positive cells are shown by the arrows. Scale bar: 50 µm. (C–E) Dual immunofluorescence staining for caspase‐3 (C)/GSDMD (D)/RIPK3 (E) (red), F4/80 (green), and DAPI (blue) in the aortas of ApoE −/− mice fed an HFD or normal diet demonstrate that GSDMD/caspase‐3/RIPK3 immunoreactivity colocalises with macrophage marker CD68. Scale bar: 50 µm. (F) Representative Western blots and relative quantitative analysis of NLRP3, GSDMD and GSDMD‐N in the aortas of ApoE −/− mice fed with a normal diet or HFD, NLRP3 agonist nigericin‐treated ApoE −/− mice fed with an HFD, and Galectin‐3 −/− / ApoE −/− mice fed with an HFD. (G) Representative Western blots and relative quantitative analysis of caspase‐3, cleaved caspase‐3, caspase 8 and cleaved caspase 8 in the aortas of ApoE −/− mice fed with a normal diet or HFD, nigericin‐treated ApoE −/− mice fed with HFD, and Galectin‐3 −/− / ApoE −/− mice fed with HFD. (H) The activity of caspase‐3 in the aortas of ApoE −/− mice fed with a normal diet or HFD, nigericin‐treated ApoE −/− mice fed with HFD, and Galectin‐3 −/− / ApoE −/− mice fed with an HFD. (I and J) Representative Western blots and relative quantitative analysis of RIPK3, MLKL and phospho‐MLKL in the aortas of ApoE −/− mice fed with a normal diet or HFD, nigericin‐treated ApoE −/− mice fed with HFD, and Galectin‐3 −/− / ApoE −/− mice fed with an HFD. (K and L) Representative Western blots and relative quantitative analysis of TLR4, MyD88, NF‐κB and phospho‐NF‐κB in the aortas of ApoE −/− mice fed with a normal diet or HFD, nigericin‐treated ApoE −/− mice fed with HFD, and Galectin‐3 −/− / ApoE −/− mice fed with an HFD. n = 4–8 mice per group. * p ˂.05, ** p ˂.01, *** p ˂.001 by Student's t test. ns: not significant.

Article Snippet: Galectin‐3 and NLRP3 expression levels are elevated in both human and murine atherosclerotic lesions.

Techniques: Clinical Proteomics, Membrane, Transmission Assay, Electron Microscopy, Immunofluorescence, Staining, Marker, Western Blot, Activity Assay

Journal: Clinical and Translational Medicine

Article Title: Macrophage‐derived galectin‐3 contributes to pyroptosis, apoptosis and necroptosis through TLR4/MyD88/NF‐κB/NLRP3 during atherosclerosis

doi: 10.1002/ctm2.70637

Figure Lengend Snippet: TLR4 silencing inhibits the development of atherosclerotic lesions via the downstream MyD88/NF‐kB/NLRP3 signalling molecules. (A) The mRNA level of TLR4 is markedly reduced in the atherosclerotic aorta of HFD‐fed ApoE −/− mice administered by AAV‐F4/80‐shTLR4. (B–D) Representative Western blots and relative quantitative analysis of TLR4, MyD88, NF‐kB, phospho‐NF‐kB and NLRP3 in the aortas of HFD‐fed ApoE−/− mice, empty vector‐treated HFD‐fed ApoE−/− mice and shTLR4‐treated HFD‐fed ApoE−/− mice. (E) Representative en face Oil Red O‐stained images of the entire aorta are obtained from HFD‐fed ApoE−/− mice, empty vector‐treated HFD‐fed ApoE−/− mice and shTLR4‐treated HFD‐fed ApoE−/− mice. Scale bar: 50 mm. (F) En face lesion area is quantified as a percentage of the total aortic surface area. After 16 weeks of HFD feeding, en face lesion areas are significantly smaller in HFD‐fed ApoE −/− mice treated with shTLR4 compared with those treated with the empty vector. (G) Representative sections of HE, Oil Red O and Movat's staining in the aortic sinuses are acquired from three different groups of mice. Scale bar: 1 mm. (H) Aortic sinus plaque lesion area, lipid lesion area and mucin area are represented as total area in µm 2 . Plaque area, lipid lesion area (red) and mucin area (blue‐green) are much smaller in HFD‐fed ApoE −/− mice treated with shTLR4 compared with those treated with empty vector. (I) Necrotic core area is calculated as the total area in µm 2 . Fibrous cap thickness is designated as average thickness in µm. TLR4 silencing diminishes the necrotic core area and fibrous cap thickness in vivo. (J and K) The silencing efficacy of TLR4 in ox‐LDL‐treated RAW 264.7 macrophages is verified by western blotting and RT‐ qPCR. (L and M) Representative Western blots and relative quantitative analysis of MyD88, NF‐kB, phospho‐NF‐kB and NLRP3 in ox‐LDL‐induced macrophages transduced by shTLR4 with or without co‐treatment by the NF‐κB activator berubicin. (N and O) Representative Western blots and relative quantitative analysis of MyD88, NF‐κB, phospho‐NF‐κB and NLRP3 in ox‐LDL‐induced macrophages transduced by shTLR4 with or without co‐treatment by the NLRP3 activator nigericin. n = 4–8 mice per group. Data are derived from three to five independent experiments. * p ˂.05, ** p ˂.01, *** p ˂.001 by Student's t test. ns: not significant.

Article Snippet: Galectin‐3 and NLRP3 expression levels are elevated in both human and murine atherosclerotic lesions.

Techniques: Western Blot, Plasmid Preparation, Staining, In Vivo, Quantitative RT-PCR, Derivative Assay

Journal: Clinical and Translational Medicine

Article Title: Macrophage‐derived galectin‐3 contributes to pyroptosis, apoptosis and necroptosis through TLR4/MyD88/NF‐κB/NLRP3 during atherosclerosis

doi: 10.1002/ctm2.70637

Figure Lengend Snippet: Galectin‐3 genetic deficiency or knockdown reduced and, conversely, NLRP3 agonist nigericin augmented atherosclerotic lesions in HFD‐fed ApoE −/− mice. (A) The knockout efficacy of galectin‐3 in the aorta is verified by Western blotting and RT‐qPCR. (B) Schematic diagram of animal study design. Galectin‐3 −/− /ApoE −/− mice and ApoE −/− mice are fed an HFD for 16 weeks, and ApoE −/− mice are intraperitoneally administered with NLRP3 agonist nigericin. (C) Representative images of en face Oil Red O staining in the entire aortas are obtained from ApoE −/− control mice, HFD‐fed ApoE −/− mice, HFD‐fed ApoE −/− mice treated with nigericin, and HFD‐fed Galectin‐3 −/− /ApoE −/− mice. Scale bar: 50 mm. (D) En face lesion area is quantified as a percentage of the total area of the aorta. Compared with those in HFD‐fed ApoE −/− mice, en face lesion areas are significantly smaller in HFD‐fed Galectin‐3 −/− /ApoE −/− mice and, conversely, are markedly bigger in HFD‐fed ApoE −/− mice treated with nigericin. (E) Representative sections of HE, Oil Red O and Movat's staining in the aortic sinuses are acquired from four different groups of mice. Scale bar: 1 mm. (F) Aortic sinus plaque lesion area, lipid lesion area and mucin area are represented as total area in µm 2 . Plaque area, lipid lesion area (red) and mucin area (blue‐green) are much bigger in HFD‐fed ApoE −/− mice treated with nigericin and, conversely, are relatively smaller in HFD‐fed Galectin‐3 −/− /ApoE −/− mice in comparison with HFD‐fed ApoE −/− mice. (G) The levels of inflammatory cytokines in the aortas are measured from four different groups of mice. (H) Schematic illustration of experimental protocol in ApoE −/− mice receiving the injection of AAV‐F4/80 shGalectin‐3/empty vector at the age of 4 weeks. After 2 weeks of a normal diet for rest, these mice are treated with HFD for 16 weeks. (I) The knockdown efficacy of shGalectin‐3 in the aorta is confirmed through Western blotting and RT‐qPCR. (J) Representative images of en face Oil Red O staining in the entire aortas are obtained from HFD‐fed ApoE −/− mice, empty vector‐treated HFD‐fed ApoE −/− mice and shGalectin‐3‐treated HFD‐fed ApoE −/− mice. Scale bar: 50 mm. (K) En face lesion area, quantified as a percentage of total area of the aorta, is significantly smaller in shGalectin‐3‐treated HFD‐fed ApoE −/− mice than in empty vector‐treated HFD‐fed ApoE −/− mice. (L) Representative sections of HE, Oil Red O and Movat's staining in the aortic sinuses are acquired from three different groups of mice. Scale bar: 1 mm. (M) Aortic sinus plaque lesion area, lipid lesion area (red) and mucin area (blue‐green), represented as total area in µm 2 , are much bigger in shGalctin‐3‐treated HFD‐fed ApoE −/− mice than in empty vector‐treated HFD‐fed ApoE −/− mice. (N) The levels of inflammatory cytokines (TNF‐1α, IL‐1β, IL‐18 and IL‐6) in the aortas are measured from three different groups of mice. n = 4–8 mice per group. * p ˂.05, ** p ˂.01, *** p ˂.001 by Student's t test. ns: not significant.

Article Snippet: Galectin‐3 and NLRP3 expression levels are elevated in both human and murine atherosclerotic lesions.

Techniques: Knockdown, Knock-Out, Western Blot, Quantitative RT-PCR, Staining, Control, Comparison, Injection, Plasmid Preparation

Journal: BMC Ophthalmology

Article Title: Integration of tear fluid biomarkers and machine learning for the early detection of keratoconus

doi: 10.1186/s12886-025-04451-8

Figure Lengend Snippet: Boxplots of Tear Cytokines with Significant Intergroup Differences. Boxplots illustrate the distribution of tear cytokine levels among normal controls, early KC, and KC groups. Only cytokines showing significant overall differences by the Kruskal–Wallis test ( p < 0.05) are presented. Pairwise comparisons were performed using the Mann–Whitney U test with Bonferroni correction. Statistical significance is indicated as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, and ns = not significant. Horizontal lines denote the group comparisons, and data points represent individual subjects

Article Snippet: Moreover, the levels of cytokines (IL-1β, Galectin-1, and Galectin-3) in KC and early KC were greater than those in the normal control (all P < 0.05, Fig. ).

Techniques: MANN-WHITNEY

Journal: BMC Ophthalmology

Article Title: Integration of tear fluid biomarkers and machine learning for the early detection of keratoconus

doi: 10.1186/s12886-025-04451-8

Figure Lengend Snippet: Feature Selection and Model Optimization for Early KC Prediction. The random forest model trained with all the parameters achieved a 5-fold cross-validation score of 0.868 ( a ) and an ROC curve AUC of 0.939 for predicting early KC ( b ). Feature importance analysis revealed the D-index, BE, and Kmax as key clinical parameters, whereas Galectin-3, IL-1β, and Galectin-1 were identified as critical tear fluid cytokines ( c ). A new random forest model was trained using these six selected features. This model achieved an AUC of 0.955 (normal: 0.976, KC: 1.000), outperforming the model trained with all the parameters ( d ). The confusion matrix for the optimized model demonstrated its excellent predictive accuracy, particularly in identifying KC and normal cases ( e )

Article Snippet: Moreover, the levels of cytokines (IL-1β, Galectin-1, and Galectin-3) in KC and early KC were greater than those in the normal control (all P < 0.05, Fig. ).

Techniques: Selection, Biomarker Discovery

Journal: The FASEB Journal

Article Title: BML‐111 treatment prevents cardiac apoptosis and oxidative stress in a mouse model of autoimmune myocarditis

doi: 10.1096/fj.202000611r

Figure Lengend Snippet: FIGURE 1 Hearts from BML-111 treated-mice presented lower inflammatory levels than EAM-mice and a lesser degree of immune infiltration (A) Representative images of histological analysis of immune cell infiltration in cardiac tissue from mice of each experimental group and the corresponding quantification (B) following Dallas criteria. Boxes represent median and interquartile range; whiskers represent maximum and minimum values. Objective lens 20X. n ≥ 20. C, Cardiac mRNA levels of several pro-inflammatory mediators (Il1b, Il6 and Tnfa) determined by qPCR. Graphs represent fold induction (FI) of each experimental group using Rplp0 as endogenous control ± SD. n ≥ 11 (Il1b), n ≥ 13 (Il6), n ≥ 18 (Tnfa). D, Representative western blot images (left panel) and their corresponding quantification (right panel) of Galectin-3 and TNFα protein levels in cardiac tissue from all experimental groups. Normalization was performed using GAPDH as reference. Graphs represent mean ± SD. n ≥ 7 (Galectin-3), n ≥ 6 (TNFα). Statistical significance was determined using one-way ANOVA followed by Tukey's multiple comparisons test for multiple groups comparisons. **P ≤ .01 and ***P ≤ .001 vs Control + Veh; #P ≤ .05 and ##P ≤ .01 vs EAM + Veh group

Article Snippet: D, Representative western blot images (left panel) and their corresponding quantification (right panel) of Galectin-3 and TNFα protein levels in cardiac tissue from all experimental groups.

Techniques: Control, Western Blot