Journal: Bioactive Materials

Article Title: 3D-MSCs apoptotic bodies-integrated conductive hydrogel mitigates spinal cord injury via immunoregulation and alleviating neuronal pyroptosis

doi: 10.1016/j.bioactmat.2026.01.043

Figure Lengend Snippet: Composite hydrogel promotes the polarization of BV2 cells to M2 types and alleviates PC12 cell pyroptosis in vitro inflammatory environment. (A) Representative western blots showing protein expression of iNOS and Arg-1 in each group, β-actin was utilized as a loading control. (B) Quantitative analysis of relative expression of iNOS and Arg-1. (C) Representative immunofluorescence images of CD68 positive and iNOS positive BV2 cells (scale bar: 20 μm). (D) Representative immunofluorescence images of CD68 positive and Arg-1 positive BV2 cells (scale bar: 20 μm). (E, F) Quantitative analysis of relative fluorescence intensity of iNOS and Arg-1. (G) Representative western blots showing the expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein associated with pyroptosis, β-actin was utilized as a loading control. (H) PI staining of PC12 cells in each group (scale bar, 100 μm). (I) Quantitative analysis of PI staining of PC12 cells (n = 3). (J) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA): The protein levels of IL-18 (Servicebio, Wuhan, China) and IL-1β (Servicebio, Wuhan, China) in the spinal cord tissue were detected using ELISA kits according to the manufacturer's protocol.

Techniques: In Vitro, Western Blot, Expressing, Control, Immunofluorescence, Fluorescence, Staining, Comparison

Journal: Bioactive Materials

Article Title: 3D-MSCs apoptotic bodies-integrated conductive hydrogel mitigates spinal cord injury via immunoregulation and alleviating neuronal pyroptosis

doi: 10.1016/j.bioactmat.2026.01.043

Figure Lengend Snippet: The composite hydrogel inhibits post-SCI pyroptosis in neurons. (A) Immunofluorescence images of residual neurons existing in the anterior horn of the spinal cord (scale bar, 500 μm and 200 μm). (B) Immunofluorescence image of Caspase-1 expression of neurons 3 days after SCI (scale bar, 20 μm). (C) Quantitative analysis of relative fluorescence intensity of Caspase-1 protein expression in neurons within the specified groups (n = 3). (D) Immunofluorescence image of GSDMD-N expression of neurons 3 days after SCI (scale bar, 20 μm). (E) Quantitative analysis of relative fluorescence intensity of GSDMD-N protein expression in neurons within the specified groups (n = 3). (F) Representative western blots showing the expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein 3 days after SCI, GAPDH was utilized as a loading control. (G) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA): The protein levels of IL-18 (Servicebio, Wuhan, China) and IL-1β (Servicebio, Wuhan, China) in the spinal cord tissue were detected using ELISA kits according to the manufacturer's protocol.

Techniques: Immunofluorescence, Expressing, Fluorescence, Western Blot, Control, Comparison

Journal: Research

Article Title: Neuromodulation and Copper Chelation Reverse Sleep Fragmentation-Aggravated Myocardial Ischemia–Reperfusion Injury by Targeting NET-Induced Endothelial Cuproptosis

doi: 10.34133/research.1266

Figure Lengend Snippet: Chronic SF exacerbated the inflammatory response and promoted the formation of NETs in MI/RI mice. (A to C) Plasma concentrations of IL-1β, IL-6, and IL-18 in mice were measured by ELISA ( n = 8 per group). (D to F) qRT-PCR assays were performed to determine mRNA levels of IL-1β, IL-6, and IL-18 in the hearts of MI/RI mice with SF ( n = 6 per group). (G) Representative images of immunofluorescence staining of NETs in myocardial tissues from Sham, SF + Sham, MI/RI, and SF + MI/RI groups (green: MPO, red: citH3, blue: DAPI) ( n = 6 per group). (H) Cell-free DNA (cfDNA) of each group ( n = 6 per group). (I) The MPO–DNA complex relative levels were assessed in mice ( n = 6 per group). Data are presented as mean ± SD. ### P < 0.001 versus Sham group; *** P < 0.001 versus SF + Sham group; $$ P < 0.01 versus MI/RI group; $$$ P < 0.001 versus MI/RI group. (J and K) Western blotting was used to analyze the expression of CXCR2 in NEs after treatment with EPI for 6 h ( n = 6 per group). Data are presented as mean ± SD. ### P < 0.001 versus Control group. C + EPI, Control + EPI. (L) Schematic diagram of coculture of EPI-induced NEs and CMECs exposed to H/R. CMECs were treated with hypoxia for 12 h and reoxygenation for 24 h and then cocultured with NEs for 6 h. (M) Statistical analysis of the number of migrated NEs after coculture with H/R-induced CMECs ( n = 6 independent experiments). (N and O) The cfDNA and MPO–DNA complex levels were assessed ( n = 5 independent experiments). (P) After coculture, the ultrastructure of NEs was observed via the electron microscope. Data are presented as mean ± SD. ### P < 0.001 versus Control + NEs group; *** P < 0.001 versus H/R + NEs group. Scale bars, 50 μm (G) and 2.5 μm (P). NEs, neutrophils; EPI, epinephrine.

Article Snippet: The plasma concentrations of IL-1β (EM0029, HUABIO), IL-18 (EM0034, HUABIO), and IL-6 (EM0004, HUABIO) were determined using ELISA kits per the supplier’s protocol.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Immunofluorescence, Staining, Western Blot, Expressing, Control, Microscopy

Journal: PLOS One

Article Title: In vivo mouse model of calcific myonecrosis induced by injury

doi: 10.1371/journal.pone.0346816

Figure Lengend Snippet: (a-c) Quantitative RT-PCR analysis of Nlrp3 (a) , Asc (b) , and Caspase-1 (c) mRNA expression in skeletal muscle from control and notexin-injected mice (n = 3). Data are presented as mean ± S.D. Statistical significance was assessed using unpaired t-test (** P < 0.01, *** P < 0.001). (d) Immunostaining of gastrocnemius muscle sections 3 days after notexin injection using anti-NLRP3, anti-ASC, and anti-Caspase-1 antibodies on serial sections. The upper panels show control muscles, and the lower panels show notexin-treated muscles. Scale bars, 20 μm. Asterisks indicate calcified muscle fibers. (e) Immunofluorescence staining showing the localization of NLRP3 (green), ASC or Caspase-1 (red) around calcified muscle fibers in notexin-injected skeletal muscle. Nuclei are counterstained with DAPI (blue). Scale bar, 20 μm. Asterisks indicate calcified muscle fibers. (f, g) Quantitative RT-PCR analysis of Il-1β (f) and Il-18 (g) mRNA expression in skeletal muscle from control and notexin-injected mice (n = 3). Data are presented as mean ± S.D. Statistical significance was assessed using unpaired t-test (*** P < 0.001). (h) Serum IL-18 levels measured by ELISA in control and notexin-injected mice (n = 6 (control), n = 9 (notexin)). Data are presented as mean ± S.D. Statistical significance was assessed using unpaired t-test (* P < 0.05). (i) Quantitative RT-PCR analysis of Runx2 mRNA expression in skeletal muscle from control and notexin-injected mice (n = 3). Data are presented as mean ± S.D. Statistical significance was assessed using unpaired t-test (* P < 0.05). (j) Western blot analysis of Runx2 and tubulin (loading control) protein expression in skeletal muscle from control and notexin-injected mice.

Article Snippet: Serum IL-18 concentrations were measured using the Mouse IL-18 DuoSet ELISA Kit (DY7625-05; R&D Systems) according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Expressing, Control, Injection, Immunostaining, Muscles, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Western Blot