Journal: Bioactive Materials

Article Title: A promising magnesium-related alloy with metabolic reprogramming and antitumor effects in hepatocellular and pancreatic cancer

doi: 10.1016/j.bioactmat.2025.12.039

Figure Lengend Snippet: Metabolomic Analysis Reveals Mg and Al-Mg Induce Metabolic Reprogramming in Hepatocellular and Pancreatic Cancer Cells. (A) Metabolomic profiling of PANC-1, PANC-1-Mg, PANC-1-Al-Mg, Huh7, Huh7-Mg, and Huh7-Al-Mg groups using LC-MS identified 1824 metabolites. (B) PCA illustrating clustering among different cell groups. (C) Heatmap showing differential abundances of characteristic metabolites across cell groups. (D) K-means clustering analysis highlighting metabolic differences among the cell groups. (E) Venn diagram displaying common differential metabolites among treatment groups. (F) Volcano plots of differential metabolites following Mg or Al-Mg treatment. (G) KEGG pathway enrichment analysis of differential metabolites. (H) Enrichment distribution of differential metabolites in Huh7 or PANC-1 cells treated with Mg or Al-Mg. (I) Quantitative analysis of intracellular metabolites including L-glutamine, adenine, uridine, cytidine, and guanine by ELISA with Mg or Al-Mg exposure. (J) Western blot analysis of p21, CDK4, and PCNA expression in PANC-1 cells after Mg or Al-Mg exposure. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: After blocking with 5 % nonfat milk for 1 h at room temperature, membranes were incubated overnight at 4 °C with primary antibodies, including AMPK (1:1000, CST, 2532), p-AMPK (1:1000, CST, 2535), CPT1B (1:1000, Proteintech, 22170-1-AP), CDK4 (1:1000, Proteintech, 11026-1-AP), PCNA (1:1000, Proteintech, 10205-2-AP), p21 (1:1000, Proteintech, 10355-1-AP), GAPDH (1:1000, Proteintech, 60004-1-Ig) followed by HRP-conjugated secondary antibody (1:5000, Proteintech, RGAR001) for 1 h at room temperature.

Techniques: Liquid Chromatography with Mass Spectroscopy, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

Journal: World Journal of Diabetes

Article Title: Formononetin inhibits p53 signaling pathway activation to delay cellular senescence and ameliorates diabetic kidney disease

doi: 10.4239/wjd.v17.i2.112500

Figure Lengend Snippet: Formononetin intervention delays high glucose-induced senescence in MPC-5 cells. A: Heatmap of genes related to the p53 signaling pathway showing that formononetin (FN) intervention upregulates MDM2 and CCND1 expression and downregulates p21 expression; B: The molecular docking results of FN and MDM2; C: Western blot bands of proliferating cell nuclear antigen; D and E: Cell staining and cell supernatant β-galactosidase (β-GAL) activity assay demonstrating that FN intervention significantly reduces β-GAL activity in MPC-5 cells; F: Β-GAL indicator cell staining photography (100 ×); G: Western blot analysis revealing that FN intervention significantly decreases proliferating cell nuclear antigen protein levels. n = 3 per group. a P < 0.05 vs control groups; b P < 0.01 vs control groups; c P < 0.05 vs high glucose groups; d P < 0.01 vs high glucose groups. NS: Not significant; FN: Formononetin; HG: High glucose; H-FN: High dose of formononetin; PCNA: Proliferating cell nuclear antigen.

Article Snippet: Sections were then incubated with the following rabbit-derived primary antibodies: Anti-nephrin (Affinity, DF7501; 1:100), anti-p21 (Bioss, bs-55160R; 1:50), anti-Ki67 (Proteintech, 28074-1-AP; 1:2000), and anti-α-smooth muscle actin (Proteintech, 14395-1-AP; 1:3000).

Techniques: Expressing, Western Blot, Staining, Activity Assay, Control

Journal: World Journal of Diabetes

Article Title: Formononetin inhibits p53 signaling pathway activation to delay cellular senescence and ameliorates diabetic kidney disease

doi: 10.4239/wjd.v17.i2.112500

Figure Lengend Snippet: Formononetin intervention delays senescence of MPC-5 cells via p53 signaling pathway. A: Western blot bands of p53 signaling pathway; B: Dual-luciferase reporter gene assay demonstrating that formononetin (FN) intervention significantly reduces p53 transcriptional activity; C-F: Western blot analysis reveals that FN intervention significantly increases MDM2 and CCND1 protein levels and decreases p53 and p21 protein levels; G: Β-galactosidase (β-GAL) indicator cell staining photography (100 ×); H: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay showing that MDM2 silencing weakens the protective effects of FN on high glucose-induced MPC-5 cell viability; I and J: Cell staining and cell supernatant β-GAL activity assay showing that MDM2 silencing eliminates the reduction in β-GAL activity induced by FN. n = 3 per group. a P < 0.05 vs control groups; b P < 0.01 vs control groups; c P < 0.05 vs high glucose groups; d P < 0.01 vs high glucose groups. NS: Not significant; FN: Formononetin; HG: High glucose; H-FN: High dose of formononetin; C: Control; shNC: Short hairpin negative control; shMDM2: Short hairpin RNA targeting MDM2.

Article Snippet: Sections were then incubated with the following rabbit-derived primary antibodies: Anti-nephrin (Affinity, DF7501; 1:100), anti-p21 (Bioss, bs-55160R; 1:50), anti-Ki67 (Proteintech, 28074-1-AP; 1:2000), and anti-α-smooth muscle actin (Proteintech, 14395-1-AP; 1:3000).

Techniques: Western Blot, Luciferase, Reporter Gene Assay, Activity Assay, Staining, Control, Negative Control, shRNA

Journal: World Journal of Diabetes

Article Title: Formononetin inhibits p53 signaling pathway activation to delay cellular senescence and ameliorates diabetic kidney disease

doi: 10.4239/wjd.v17.i2.112500

Figure Lengend Snippet: MDM2 silencing attenuates anti-senescence effects of formononetin. We silenced the MDM2 gene in MPC-5 cells to investigate whether formononetin (FN) exerts its anti-senescence effects through MDM2 ( n = 3 per group). A: Western blot bands of p53 signaling pathway; B: Dual-luciferase reporter gene assay demonstrating that MDM2 silencing attenuates inhibitory effects of FN on p53 transcriptional activity; C-G: Western blot analysis showing that MDM2 silencing abolishes regulatory effects of FN on MDM2, p53, p21, proliferating cell nuclear antigen and CCND1 protein expression; H-J: Silencing MDM2 abolishes inhibitory effects of FN on interleukin-1β, interleukin-6, and tumor necrosis factor α levels in the culture supernatant of high glucose-exposed MPC-5 cells. a P < 0.05 vs control groups; b P < 0.01 vs control groups; c P < 0.05 vs high glucose groups; d P < 0.01 vs high glucose groups; e P < 0.05 vs short hairpin RNA targeting MDM2 group; f P < 0.01 vs short hairpin RNA targeting MDM2 groups. NS: Not significant; FN: Formononetin; HG: High glucose; C: Control; shNC: Short hairpin negative control; shMDM2: Short hairpin RNA targeting MDM2; PCNA: Proliferating cell nuclear antigen; IL-1β: Interleukin-1β; IL-6: Interleukin-6; TNF-α: Tumor necrosis factor α.

Article Snippet: Sections were then incubated with the following rabbit-derived primary antibodies: Anti-nephrin (Affinity, DF7501; 1:100), anti-p21 (Bioss, bs-55160R; 1:50), anti-Ki67 (Proteintech, 28074-1-AP; 1:2000), and anti-α-smooth muscle actin (Proteintech, 14395-1-AP; 1:3000).

Techniques: Western Blot, Luciferase, Reporter Gene Assay, Activity Assay, Expressing, Control, shRNA, Negative Control

Journal: World Journal of Diabetes

Article Title: Formononetin inhibits p53 signaling pathway activation to delay cellular senescence and ameliorates diabetic kidney disease

doi: 10.4239/wjd.v17.i2.112500

Figure Lengend Snippet: Formononetin intervention ameliorates podocyte injury and cellular senescence in diabetic kidney disease mice. A: Immunofluorescence staining of podocyte-specific markers (nephrin, podocin, and CD2AP; bars = 50 μm); B: Immunofluorescence staining of fibrosis markers (α-smooth muscle actin; bars = 50 μm); C: Immunofluorescence staining of aging markers (p21; bars = 50 μm); D: Immunofluorescence staining of cellular proliferation dynamics markers (Ki67; bars = 50 μm). n = 4 per group. a P < 0.05 vs control groups; b P < 0.01 vs high glucose groups. NS: Not significant; IRB: Irbesartan; L-FN: Low dose of formononetin; M-FN: Medium dose of formononetin; H-FN: High dose of formononetin; α-SMA: Α-smooth muscle actin.

Article Snippet: Sections were then incubated with the following rabbit-derived primary antibodies: Anti-nephrin (Affinity, DF7501; 1:100), anti-p21 (Bioss, bs-55160R; 1:50), anti-Ki67 (Proteintech, 28074-1-AP; 1:2000), and anti-α-smooth muscle actin (Proteintech, 14395-1-AP; 1:3000).

Techniques: Immunofluorescence, Staining, Control

Journal: World Journal of Diabetes

Article Title: Formononetin inhibits p53 signaling pathway activation to delay cellular senescence and ameliorates diabetic kidney disease

doi: 10.4239/wjd.v17.i2.112500

Figure Lengend Snippet: Formononetin exhibits anti-senescence effects in diabetic kidney disease mice. A-C: Formononetin (FN) intervention significantly reduces levels of interleukin-1β, interleukin-6, and tumor necrosis factor α in renal tissues of diabetic kidney disease mice; D: FN intervention significantly decreases β-galactosidase activity in renal tissues; E-H: FN intervention significantly upregulates MDM2 and CCND1 gene expression and downregulates p53 and p21 gene expression; I: Western blot bands of p53 signaling pathway; J-M: FN intervention significantly increases MDM2 and CCND1 protein levels and decreases p53 and p21 protein levels. n = 10 for A-D, n = 3 for F. a P < 0.05 vs control groups; b P < 0.01 vs control groups; c P < 0.05 vs high glucose groups; d P < 0.01 vs high glucose groups. H-FN: High dose of formononetin; IL-1β: Interleukin-1β; IL-6: Interleukin-6; TNF-α: Tumor necrosis factor α.

Article Snippet: Sections were then incubated with the following rabbit-derived primary antibodies: Anti-nephrin (Affinity, DF7501; 1:100), anti-p21 (Bioss, bs-55160R; 1:50), anti-Ki67 (Proteintech, 28074-1-AP; 1:2000), and anti-α-smooth muscle actin (Proteintech, 14395-1-AP; 1:3000).

Techniques: Activity Assay, Gene Expression, Western Blot, Control

Journal: Biology

Article Title: Force-Dependent Presence of Senescent Cells Expressing Vascular Endothelial Growth Factor During Orthodontic Tooth Movement

doi: 10.3390/biology15020187

Figure Lengend Snippet: Presence and distribution of senescent cells during OTM. ( A ) Representative p21 and p16 immunofluorescence staining images with DAPI. ( B ) Quantitative analysis of p21 + area in PDL. ( C ) Quantitative analysis of p16 + area in PDL. ( D , E ) Distribution of senescence marker–positive areas in the 60 g and 180 g groups. Scale bars: 100 µm (low magnification), 10 µm (high magnification). Data are mean ± SD (n = 4). * p < 0.05; *** p < 0.001; **** p < 0.0001; one-way ANOVA with Tukey’s test. PDL: periodontal ligament, AB: alveolar bone.

Article Snippet: p21 , Senescence marker , Bioss Antibodies (Shanghai, China) , bs-10129R , AF555 , 1:100.

Techniques: Immunofluorescence, Staining, Marker

Journal: Biology

Article Title: Force-Dependent Presence of Senescent Cells Expressing Vascular Endothelial Growth Factor During Orthodontic Tooth Movement

doi: 10.3390/biology15020187

Figure Lengend Snippet: Cellular senescence of vascular endothelial cells (ECs) induced by OTM. ( A ) Representative p21 and CD31 immunofluorescence staining images. p21/CD31 staining without DAPI ( left ) and merged images with DAPI ( right ) are shown. ( B ) Quantitative analysis of p21 + CD31 + area in PDL. ( C ) Representative p16 and CD31 immunofluorescence staining images. p16/CD31 staining without DAPI ( left ) and merged images with DAPI ( right ) are shown. ( D ) Quantitative analysis of p16 + CD31 + area in PDL. Scale bars: 100 µm (low magnification), 10 µm (high magnification). Data are mean ± SD (n = 4). ns: not significant, * p < 0.05; ** p < 0.01; Kruskal–Wallis with Dunn’s post hoc test. PDL: periodontal ligament, AB: alveolar bone.

Article Snippet: p21 , Senescence marker , Bioss Antibodies (Shanghai, China) , bs-10129R , AF555 , 1:100.

Techniques: Immunofluorescence, Staining