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: Transcriptomic Analysis Indicates Mg and Al-Mg Alter Gene Expression Patterns in Hepatocellular and Pancreatic Cancer Cells. (A) High-throughput sequencing of PANC-1, PANC-1-Mg, PANC-1-Al-Mg, Huh7, Huh7-Mg, and Huh7-Al-Mg groups; heatmap showing sample correlations. (B) Volcano plots illustrating differential gene expression in Huh7 and PANC-1 cells after Mg or Al-Mg treatment. (C) GO enrichment analysis of differentially expressed genes following Mg or Al-Mg treatment. (D) KEGG pathway enrichment analysis of differentially expressed genes after Mg or Al-Mg treatment. (E) GSEA enrichment analysis of gene expression profiles post Mg or Al-Mg treatment. (F) Western blot analysis of AMPK, p-AMPK and CPT1B expression in PANC-1 cells after Mg or Al-Mg exposure with or without AMPK agonist. (G) PPI network analysis of differentially expressed genes in Mg or Al-Mg-treated cells. (H) Integrated analysis of metabolomic and transcriptomic data by MetaboAnalyst 6.0.

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: Gene Expression, Next-Generation Sequencing, Western Blot, Expressing

Journal: Bioactive Materials

Article Title: An intelligent nanoliposome alleviates disc degeneration and discogenic pain by inhibiting neurovascular ingrowth via a “Soil-conditioning, seed-modulating, and weeds-suppressing” strategy

doi: 10.1016/j.bioactmat.2025.11.048

Figure Lengend Snippet: Oxidative stress and aberrant neurovascularization are progressively enhanced during IVDD. (A) Representative T2-weighted MRI images showing disc structure in human intervertebral discs at varying Pfirrmann degeneration grades (II–IV). (B–C) Fluorescent staining and quantitative analysis of ROS levels in primary NPCs from grades II–IV NP tissues, showing a significant increase in oxidative stress with disease severity (n = 5). Scale bar, 20 μm. (D–F) IHC staining and quantitative analyses of CD31 and Tuj1 across different degenerative grades (n = 5). Scale bar, 50 μm. (G) H&E, SOFG, and IF staining of COL2A1 and MMP3 for IVDs (n = 5). Scale bar, 100 μm. (H) Histological score of the IVD tissues (n = 5). (I–J) Quantification of IF staining for COL2A1 and MMP3 (n = 5). (K–M) IF staining and corresponding quantification analyses of CD31 and Tuj1 in disc tissues of sham and AFP-treated mice (n = 5). Scale bar, 100 μm. Data are presented as mean ± SD. The comparisons among groups were performed using one-way ANOVA. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; ns = not significant.

Article Snippet: For immunofluorescence (IF) staining, antigen retrieval was conducted using citrate buffer (0.1 M, pH 6.0), followed by blocking in 10 % normal goat serum (Solarbio, Cat. No. SL038) at room temperature for 1 h. Sections were incubated overnight at 4 °C with primary antibodies targeting ACAN (Cat. No. DF7561, Affinity), COL2A1 (Cat. No. AF0135, Affinity), MMP3 (Cat. No. AF0217, Affinity), MMP13 (Cat. No. 18165-1-AP, Proteintech), CD31 (Cat. No. AF0077, Affinity), p-LATS1 (Cat. No. AF7170, Affinity), p-YAP (Cat. No. AF3328, Affinity), Tuj1 (Cat. No. 66375, Proteintech), CGRP (Cat. No. DF7386, Affinity), SP (Cat. No. DF7522, Affinity), and PGP9.5 (Cat. No. AF5490, Affinity) followed by incubation with appropriate fluorescent secondary antibodies for 2 h at room temperature and counterstaining with DAPI (Beyotime, Cat. No. C1006).

Techniques: Staining, Immunohistochemistry

Journal: Bioactive Materials

Article Title: An intelligent nanoliposome alleviates disc degeneration and discogenic pain by inhibiting neurovascular ingrowth via a “Soil-conditioning, seed-modulating, and weeds-suppressing” strategy

doi: 10.1016/j.bioactmat.2025.11.048

Figure Lengend Snippet: NM-LP TK /RSV-MnCDs suppresses pathological neurovascularization in IVDD. (A–B) Representative IF images and quantitative analyses of CD31 and Tuj1 in disc regions from different groups (n = 5). Scale bar, 100 μm. (C) Schematic of the indirect co-culture Transwell system used to evaluate paracrine proangiogenic signaling from NPCs to MAECs or DRG neurons. (D) Representative images of capillary-like tube formation by MAECs cultured with NPCs from different treatment groups (n = 5). Scale bar, 100 μm. (E) Quantification of number of branch points and total tube length (n = 5). (F–G) Representative wound healing images and quantitative analysis of MAECs at 0 h and 24 h post-scratch in different co-culture conditions (n = 5). Scale bar, 100 μm. (H) Schematic of DRG neurons collection and co-culture. (I–J) Representative IF images of TUBB3 and quantification of neurites length in DRG neurons. Scale bar, 20 μm. Data are presented as mean ± SD. The comparisons among groups were performed using one-way ANOVA. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; ns = not significant.

Article Snippet: For immunofluorescence (IF) staining, antigen retrieval was conducted using citrate buffer (0.1 M, pH 6.0), followed by blocking in 10 % normal goat serum (Solarbio, Cat. No. SL038) at room temperature for 1 h. Sections were incubated overnight at 4 °C with primary antibodies targeting ACAN (Cat. No. DF7561, Affinity), COL2A1 (Cat. No. AF0135, Affinity), MMP3 (Cat. No. AF0217, Affinity), MMP13 (Cat. No. 18165-1-AP, Proteintech), CD31 (Cat. No. AF0077, Affinity), p-LATS1 (Cat. No. AF7170, Affinity), p-YAP (Cat. No. AF3328, Affinity), Tuj1 (Cat. No. 66375, Proteintech), CGRP (Cat. No. DF7386, Affinity), SP (Cat. No. DF7522, Affinity), and PGP9.5 (Cat. No. AF5490, Affinity) followed by incubation with appropriate fluorescent secondary antibodies for 2 h at room temperature and counterstaining with DAPI (Beyotime, Cat. No. C1006).

Techniques: Co-Culture Assay, Cell Culture

Journal: Bioactive Materials

Article Title: An intelligent nanoliposome alleviates disc degeneration and discogenic pain by inhibiting neurovascular ingrowth via a “Soil-conditioning, seed-modulating, and weeds-suppressing” strategy

doi: 10.1016/j.bioactmat.2025.11.048

Figure Lengend Snippet: NM-LP TK /RSV-MnCDs suppresses vascular ingrowth and matrix degradation in AFP-induced disc degeneration through Hippo pathway inactivation. (A–D) Representative IF images and quantitative analyses of CD31 and Tuj1 in IVDs from AFP-induced IVDD mice treated with MCB, TRULI, or their combinations with NM-LP TK /RSV-MnCDs (n = 5). Scale bar, 100 μm. (E–H) IF staining and quantification of COL2A1 and MMP13 expression in NP regions (n = 5). Scale bar, 100 μm. Data are presented as mean ± SD. The comparisons among groups were performed using one-way ANOVA. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; ns = not significant.

Article Snippet: For immunofluorescence (IF) staining, antigen retrieval was conducted using citrate buffer (0.1 M, pH 6.0), followed by blocking in 10 % normal goat serum (Solarbio, Cat. No. SL038) at room temperature for 1 h. Sections were incubated overnight at 4 °C with primary antibodies targeting ACAN (Cat. No. DF7561, Affinity), COL2A1 (Cat. No. AF0135, Affinity), MMP3 (Cat. No. AF0217, Affinity), MMP13 (Cat. No. 18165-1-AP, Proteintech), CD31 (Cat. No. AF0077, Affinity), p-LATS1 (Cat. No. AF7170, Affinity), p-YAP (Cat. No. AF3328, Affinity), Tuj1 (Cat. No. 66375, Proteintech), CGRP (Cat. No. DF7386, Affinity), SP (Cat. No. DF7522, Affinity), and PGP9.5 (Cat. No. AF5490, Affinity) followed by incubation with appropriate fluorescent secondary antibodies for 2 h at room temperature and counterstaining with DAPI (Beyotime, Cat. No. C1006).

Techniques: Staining, Expressing

Journal: Bioactive Materials

Article Title: An intelligent nanoliposome alleviates disc degeneration and discogenic pain by inhibiting neurovascular ingrowth via a “Soil-conditioning, seed-modulating, and weeds-suppressing” strategy

doi: 10.1016/j.bioactmat.2025.11.048

Figure Lengend Snippet: NM-LP TK /RSV-MnCDs alleviates neurovascular ingrowth. (A) GO enrichment analysis of RNA-seq data showing significant enrichment of axon guidance and angiogenesis signaling pathways. (B–C) GSEA plot demonstrating negative enrichment of both cell communication and axon guidance pathways after NM-LP TK /RSV-MnCDs treatment. (D) The schematic of the experimental design. (E–F) Representative IF images and quantification analyses of CD31 + Tuj1 + double-positive tissues at the cartilage endplate (n = 5). Scale bar, 100 μm. (G) Colocalization analysis of CD31 positive vessels and Tuj1 positive nerves (n = 5). Data are presented as mean ± SD. The comparisons among groups were performed using one-way ANOVA. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; ns = not significant.

Article Snippet: For immunofluorescence (IF) staining, antigen retrieval was conducted using citrate buffer (0.1 M, pH 6.0), followed by blocking in 10 % normal goat serum (Solarbio, Cat. No. SL038) at room temperature for 1 h. Sections were incubated overnight at 4 °C with primary antibodies targeting ACAN (Cat. No. DF7561, Affinity), COL2A1 (Cat. No. AF0135, Affinity), MMP3 (Cat. No. AF0217, Affinity), MMP13 (Cat. No. 18165-1-AP, Proteintech), CD31 (Cat. No. AF0077, Affinity), p-LATS1 (Cat. No. AF7170, Affinity), p-YAP (Cat. No. AF3328, Affinity), Tuj1 (Cat. No. 66375, Proteintech), CGRP (Cat. No. DF7386, Affinity), SP (Cat. No. DF7522, Affinity), and PGP9.5 (Cat. No. AF5490, Affinity) followed by incubation with appropriate fluorescent secondary antibodies for 2 h at room temperature and counterstaining with DAPI (Beyotime, Cat. No. C1006).

Techniques: RNA Sequencing, Protein-Protein interactions

Journal: Non-coding RNA Research

Article Title: The regulatory role of circGDI2 in hepatocellular carcinoma proliferation and glycolysis with the involvement of m6A modification

doi: 10.1016/j.ncrna.2025.11.006

Figure Lengend Snippet: Silencing circGDI2 inhibits proliferation and glycolysis in HCC cells To explore the effect of circGDI2 on proliferation and glycolysis in HCC cells, sh-circGDI2 was transfected into Li-7 and Huh-7 cells. (A) The transfection efficiency was confirmed by RT-qPCR. (B) CCK-8 assay was used to assess cell proliferation. (C) Glucose consumption level and lactate production were detected using the Glucose Assay Kit with O-toluidine and Lactate Assay Kit. (D) RT-qPCR was used to analyze the expression changes of key glycolysis-related genes (HIF-1α. HK1, PKM2, PFKM, HK2, GLUT1, PFKP, LDHA) in Li-7 and Huh-7 cell lines after circGDI2 knockdown. (E) The expression level of PKM2 was examined in clinical HCC tissues and adjacent normal tissues using RT-qPCR. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, vs. the sh-NC group, or the Adjacent group.

Article Snippet: PKM2 , Western blot , Rabbit , 1:5000 , Proteintech , 15822-1-AP.

Techniques: Transfection, Quantitative RT-PCR, CCK-8 Assay, Glucose Assay, Lactate Assay, Expressing, Knockdown

Journal: Non-coding RNA Research

Article Title: The regulatory role of circGDI2 in hepatocellular carcinoma proliferation and glycolysis with the involvement of m6A modification

doi: 10.1016/j.ncrna.2025.11.006

Figure Lengend Snippet: Silencing circGDI2 inhibits proliferation and glycolysis and PKM2 expression through IGF2BP2 in HCC cells. (A) Western blot was used to analyze IGF2BP2 expression in Li-7 and Huh-7 cell lines after circGDI2 knockdown. (B) The expression level of IGF2BP2 was examined in clinical HCC tissues and adjacent normal tissues using RT-qPCR. To clarify if circGDI2 regulated HCC cell proliferation and glycolysis through IGF2BP2, sh-circGDI2 and OE-IGF2BP2 were co-transfected into Li-7 and Huh-7 cells. (C) Western blot was used to assess the effect of IGF2BP2 on the expression of PKM2. (D) CCK-8 assay was used to assess cell proliferation. (E) Glucose consumption level and lactate production were detected using the Glucose Assay Kit with O-toluidine and Lactate Assay Kit. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, vs. the sh-NC group, or the Adjacent group. # P < 0.05, ## P < 0.01, ### P < 0.001, vs. the sh-circGDI2+OE-NC group.

Article Snippet: PKM2 , Western blot , Rabbit , 1:5000 , Proteintech , 15822-1-AP.

Techniques: Expressing, Western Blot, Knockdown, Quantitative RT-PCR, Transfection, CCK-8 Assay, Glucose Assay, Lactate Assay

Journal: Non-coding RNA Research

Article Title: The regulatory role of circGDI2 in hepatocellular carcinoma proliferation and glycolysis with the involvement of m6A modification

doi: 10.1016/j.ncrna.2025.11.006

Figure Lengend Snippet: Silencing circGDI2 inhibits HCC tumor growth and PKM2 expression through IGF2BP2. To verify the effect of circGDI2 and IGF2BP2 on HCC tumor growth, a xenograft mouse model was constructed. (A) Pictures of the isolated tumors of the indicated group. (B) The tumor volume and weight were recorded. (C) HE staining, Tunel and Ki-67 staining were performed to observe the histological characteristics and cell proliferation in the tumor tissues (scale bar = 100 μm). (D) RT-qPCR was used to detect circGDI2, IGF2BP2 and PKM2 levels. (E) IHC was used to detect IGF2BP2 and PKM2 levels (scale bar = 100 μm). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, vs. the sh-NC group. # P < 0.05, ## P < 0.01, ### P < 0.001, vs. the sh-circGDI2+OE-NC group.

Article Snippet: PKM2 , Western blot , Rabbit , 1:5000 , Proteintech , 15822-1-AP.

Techniques: Expressing, Construct, Isolation, Staining, TUNEL Assay, Quantitative RT-PCR

Journal: Non-coding RNA Research

Article Title: The regulatory role of circGDI2 in hepatocellular carcinoma proliferation and glycolysis with the involvement of m6A modification

doi: 10.1016/j.ncrna.2025.11.006

Figure Lengend Snippet: Silencing FTO inhibits HCC tumor growth and decreases circRNA, IGF2BP2 and PKM2 levels. To investigate the biological role of FTO on HCC tumor growth, the xenograft tumor models of HCC cells in the sh-NC and sh-FTO groups were established. (A) Pictures of the isolated tumors of the indicated group. (B) The tumor volume and weight were recorded. (C) HE staining, Ki-67 staining and Tunel stainning were performed to observe the histological characteristics and cell proliferation in the tumor tissues (scale bar = 100 μm). (D) RT-qPCR was used to detect circGDI2 and FTO levels. (E) IHC was used to detect IGF2BP2 and PKM2 levels (scale bar = 100 μm). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, vs. the sh-NC group.

Article Snippet: PKM2 , Western blot , Rabbit , 1:5000 , Proteintech , 15822-1-AP.

Techniques: Isolation, Staining, TUNEL Assay, Quantitative RT-PCR