Journal: BMC Gastroenterology

Article Title: Inhibition of TLR4 enhances oxaliplatin chemotherapy sensitivity in esophageal squamous cell carcinoma by suppressing inflammation and glycolysis

doi: 10.1186/s12876-026-04663-2

Figure Lengend Snippet: OXA activated TLR4 signaling in ESCC cells. The cells were treated by OXA (25 µM) for 24 h. A , B . The mRNA expressions of TLR4 and MYD88 in ESCC cell lines and normal esophageal cells were detected by qRT-PCR. C . ICC showed the immunocytochemical activity of NF-κB p65, p-NF-κB p65, COX-2, and MYD88. Scale bar: 50 μm. D . qRT-PCR analysis of mRNA levels of IL-1β, IL-6, COX-2, CXCL5, and CXCL8. *p < 0.05, **p < 0.01, ***p < 0.001 vs. Control group

Article Snippet: Serum concentrations of IL-6 (JL20268, Jonin) and IL-1β (KE10003, Proteintech) were quantified using commercial ELISA kits according to manufacturers’ protocols.

Techniques: Quantitative RT-PCR, Activity Assay, Control

Journal: BMC Gastroenterology

Article Title: Inhibition of TLR4 enhances oxaliplatin chemotherapy sensitivity in esophageal squamous cell carcinoma by suppressing inflammation and glycolysis

doi: 10.1186/s12876-026-04663-2

Figure Lengend Snippet: TLR4 knockout enhances the sensitivity of OXA chemotherapy in ESCC in vivo. A . Number of tumors. B . HE staining of esophageal epithelial tissues. Scale bar: 50 μm. C . Body weight of mice. D , E . the levels of serum IL-1β and IL-6 in mice from different groups were detected by ELISA. F . The immunohistochemical activities of PCNA, CK14, Cyclin D1, COX-2, S100A8 and S100A9 in the esophageal tissue were detected. Scale bar: 100 μm. G , H . The mRNA levels of inflammatory cytokines and glycolysis-related proteins in esophageal tissue were detected by qRT-PCR. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. WT group; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. WT + 4NQO + OXA group

Article Snippet: Serum concentrations of IL-6 (JL20268, Jonin) and IL-1β (KE10003, Proteintech) were quantified using commercial ELISA kits according to manufacturers’ protocols.

Techniques: Knock-Out, In Vivo, Staining, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Quantitative RT-PCR

Journal: BMC Gastroenterology

Article Title: Inhibition of TLR4 enhances oxaliplatin chemotherapy sensitivity in esophageal squamous cell carcinoma by suppressing inflammation and glycolysis

doi: 10.1186/s12876-026-04663-2

Figure Lengend Snippet: Schematic diagram illustrating the mechanism by which TLR4 inhibition enhances oxaliplatin (OXA) chemosensitivity in esophageal squamous cell carcinoma (ESCC). OXA treatment upregulates TLR4 and its downstream adaptor protein MYD88, which activates the phosphorylation of NF-κB p65. This activation drives two parallel pathways: (1) the inflammatory response, characterized by the upregulation of pro-inflammatory factors such as IL-6, COX-2, and CXCL5; (2) the glycolytic metabolic reprogramming, mediated by the HIF-1α/GLUT1 axis and enhanced expression of glycolytic enzymes including PFKM and LDHB. These two pathways synergistically promote ESCC cell proliferation, migration, and invasion, ultimately reducing OXA chemosensitivity. Inhibition of TLR4 (via genetic knockout, shRNA knockdown, or pharmacological inhibitor TAK-242) or its downstream mediator MYD88 (via shRNA knockdown or inhibitor ST2825) blocks NF-κB p65 phosphorylation, thereby suppressing both the inflammatory response and glycolytic activity. This dual inhibition disrupts the adaptive survival mechanisms of ESCC cells, potentiating the anti-tumor efficacy of OXA

Article Snippet: Serum concentrations of IL-6 (JL20268, Jonin) and IL-1β (KE10003, Proteintech) were quantified using commercial ELISA kits according to manufacturers’ protocols.

Techniques: Inhibition, Phospho-proteomics, Activation Assay, Expressing, Migration, Knock-Out, shRNA, Knockdown, Activity Assay

Journal: Microorganisms

Article Title: Polyethylene Terephthalate Hydrolases in Human Gut Microbiota and Their Implications for Human Health

doi: 10.3390/microorganisms12010138

Figure Lengend Snippet: Effects of PET MPs and hydrolysates on cell viability and macrophage polarization. ( A , B ) Relative expression levels of IL-1β, IL-6, TNF-α, and Arg-1 (IL-4 stimulation) in mouse monocyte macrophages RAW 264.7 cells treated with TPA (0.01 mM) ( A ) and TPA (1.00 mM) ( B ) for 24 h followed by LPS/IL-4 stimulation for 6 h. ( C ) Relative expression levels of IL-1β, IL-6, TNF-α, and Arg-1 (IL-4 stimulation) in mouse bone-marrow-derived macrophages (BMDMs) treated with TPA (1.00 mM) for 24 h followed by LPS/IL-4 stimulation for 6 h. ( D , E ) Relative expression levels of IL-1β ( D ) and Arg-1 ( E ) in RAW 264.7 cells treated with PET MPs (<400 μm) at a gradient dose at 0.01, 0.10, 0.50, and 1.00 mg/mL for 24 h followed by LPS/IL-4 stimulation for 6 h. ( F , G ) Relative expression levels of IL-1β ( F ) and Arg-1 ( G ) in RAW 264.7 cells treated with PET MPs (<400 μm) at a gradient dose at 0.01, 0.10, 0.50, and 1.00 mg/mL for 24 h without any stimulation. *** p -value < 0.001; ** p -value < 0.01; * p -value < 0.05.

Article Snippet: Macrophage colony-stimulating factor (M-CSF) (product code: HY-P7085), lipopolysaccharides (LPS) (product code: HY-D1056), and murine interleukin-4 (IL-4) (product code: HY-P70644) were obtained from MedChemExpress, Princeton, NJ, USA.

Techniques: Expressing, Derivative Assay

Journal: Experimental & Molecular Medicine

Article Title: Local delivery of OSK factors enables partial cellular reprogramming to mitigate osteoarthritis and cartilage fibrosis

doi: 10.1038/s12276-026-01662-x

Figure Lengend Snippet: a , b RT–qPCR analysis of Oct4 , Sox2 , klf4 and c-Myc ( a ) and Sox9 , Nanog ( b ) in primary chondrocytes receiving either AAV-Mock or AAV-OSK treatments. n = 3 per group. c RT–qPCR analysis of inflammatory genes ( Tnf , Il-1α and Il-1β ) and metabolic genes ( Acan , Col2 and Mmp13 ) in ATDC5 cell line under inflammatory environment induced by IL-1β for 24 h, after treatment with vehicle, AAV-Mock or AAV-OSK for 3 days. n = 3 per group. d , e ICC staining of SOX2 and IL-1β in ATDC5 cells under inflammatory conditions and different treatments ( d ) and the percentage of IL-1β positive cells ( e ). n = 3 per group. Scale bar, 50 μm. f Flow cytometry assay of apoptosis in ATDC5 cells induced by TNF, treated with vehicle, AAV-Mock or AAV-OSK. g Quantification of the ratios of apoptotic (Annexin V + /PI + ) cells in f . n = 3 per group. Data are presented as mean ± s.d. and analyzed using one-way ANOVA with Bonferroni post hoc test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: To simulate the inflammatory infiltration observed in OA conditions, ATDC5 cells were exposed to 10 ng/ml interleukin-1β (IL-1β) recombinant protein (Ag23461, Proteintech).

Techniques: Quantitative RT-PCR, Staining, Flow Cytometry

Journal: Journal of Cardiovascular Translational Research

Article Title: Bufalin Ameliorates Myocardial Ischemia/Reperfusion Injury by Suppressing Macrophage Pyroptosis via P62 Pathway

doi: 10.1007/s12265-024-10577-9

Figure Lengend Snippet: Bufalin attenuated macrophage pyroptosis and reduced IL-1β release into the myocardium and serum during I/R injury. A-C The RNA sequencing data were extracted from GSE 126772. The mRNA levels of Nlrp3 , Pycard ( ASC ) and Il-1b in macrophages isolated from heart after MI were shown as fold change compared to the Pre group (control group, n = 2,3). D Macrophages were identified and isolated from the heart by flow cytometry. E–F Immunoblotting was performed to assess the expression levels of NLRP3, GSDMD and IL-1β in macrophages after I/R injury with or without bufalin treatment. Each target band was normalized to the β-actin band and the relative expression was calculated as the fold change compared to the Sham + PBS group (control group, n = 3). G Representative IL-1β expression and localization in murine hearts are shown by IHC analysis in longitudinal sections of whole hearts. For each mouse, 5 representative zones in the left ventricle were chosen randomly to determine the percentage of IL-1β-positive ratio by ImageJ ( n = 3). H IL-1β levels in the peripheral blood of mice that were subjected to I/R injury and were treated with or without bufalin were measured by ELISA ( n = 6). IHC (immunohistochemistry). Data are expressed as the mean ± SEM. P values were determined by ordinary one-way ANOVA with Dunnett’s multiple comparisons test ( A - C ) and two-way ANOVA with Tukey’s multiple comparisons test ( F and H )

Article Snippet: For interleukin-1β (IL-1β) treatment, recombinant mouse IL-1β (HY-P7073, MedChemExpress, New Jersey, USA) was added to HL-1 cells at different concentrations (0, 100 pg/ml, 300 pg/ml, 1000 pg/ml) and incubated for 24 h. For anakinra treatment, the recombinant human interleukin-1 receptor (IL-1R) antagonist anakinra (HY-108841, MedChemExpress) was added to cardiomyocytes every 6 h for 24 h when they were cocultured with normoxia-treated or H/R-treated macrophages.

Techniques: RNA Sequencing, Isolation, Control, Flow Cytometry, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Immunohistochemistry

Journal: Journal of Cardiovascular Translational Research

Article Title: Bufalin Ameliorates Myocardial Ischemia/Reperfusion Injury by Suppressing Macrophage Pyroptosis via P62 Pathway

doi: 10.1007/s12265-024-10577-9

Figure Lengend Snippet: Bufalin attenuated H/R-treated macrophage pyroptosis in vitro. Macrophages were subjected to 1 h of hypoxia and 24 h of reoxygenation treated with PBS or bufalin. A-B Proteins were extracted from cells and immunoblotting analysis was performed. The expressions of NLRP3, GSDMD, and IL-1β were normalized to that of β-actin and then compared with the Sham + PBS group (control group, n = 3). C PI staining was used to examine the plasma membrane integrity of macrophages. Positive cells were counted and normalized to the total number of cells ( n = 5). D Cell viability was assessed by measuring the quantity of ATP in macrophages subjected to different treatments ( n = 3, 6). E IL-1β levels in the culture medium of macrophages were measured by ELISA ( n = 3). Data are presented as the mean ± SEM. P values were calculated by two-way ANOVA with Tukey’s multiple comparisons test ( B , C , and E ) or Šídák’s multiple comparisons test ( D )

Article Snippet: For interleukin-1β (IL-1β) treatment, recombinant mouse IL-1β (HY-P7073, MedChemExpress, New Jersey, USA) was added to HL-1 cells at different concentrations (0, 100 pg/ml, 300 pg/ml, 1000 pg/ml) and incubated for 24 h. For anakinra treatment, the recombinant human interleukin-1 receptor (IL-1R) antagonist anakinra (HY-108841, MedChemExpress) was added to cardiomyocytes every 6 h for 24 h when they were cocultured with normoxia-treated or H/R-treated macrophages.

Techniques: In Vitro, Western Blot, Control, Staining, Clinical Proteomics, Membrane, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cardiovascular Translational Research

Article Title: Bufalin Ameliorates Myocardial Ischemia/Reperfusion Injury by Suppressing Macrophage Pyroptosis via P62 Pathway

doi: 10.1007/s12265-024-10577-9

Figure Lengend Snippet: Bufalin restored autophagic flux by suppressing P62 pathway in H/R-induced macrophages. Macrophages were transfected with the p62-overexpression plasmid for 24 h and then subjected to H/R with or without bufalin. A - B The protein levels of P62 were measured by immunoblotting. The bands were normalized to the β-actin band, and the relative expressions were calculated as the fold changes compared to the control group ( n = 3). C The mRNA levels of p62 were measured by qRT‒PCR. The Cq results were normalized to Mus-18 s and then compared with the control group ( n = 3). D-E Proteins were extracted from H/R-treated macrophages that were infected with vector or OE-p62 plasmid in advance and treated with bufalin or PBS. Immunoblotting was performed to measure the expressions of NLRP3, GSDMD and IL-1β. Each band was normalized to the β-actin band and the relative expression was calculated as the fold change compared to the Con + PBS group (control group, n = 3). F PI staining was used to assess pyroptosis in macrophages. Positive ratios were calculated as the percentage of positive cells compared with total number of cells ( n = 5). G IL-1β levels in the culture medium of macrophages were measured by ELISA ( n = 3). Data are presented as the mean ± SEM. P values were calculated by ordinary one-way ANOVA with Tukey’s multiple comparisons test

Article Snippet: For interleukin-1β (IL-1β) treatment, recombinant mouse IL-1β (HY-P7073, MedChemExpress, New Jersey, USA) was added to HL-1 cells at different concentrations (0, 100 pg/ml, 300 pg/ml, 1000 pg/ml) and incubated for 24 h. For anakinra treatment, the recombinant human interleukin-1 receptor (IL-1R) antagonist anakinra (HY-108841, MedChemExpress) was added to cardiomyocytes every 6 h for 24 h when they were cocultured with normoxia-treated or H/R-treated macrophages.

Techniques: Transfection, Over Expression, Plasmid Preparation, Western Blot, Control, Infection, Expressing, Staining, Enzyme-linked Immunosorbent Assay

Journal: Oncotarget

Article Title: Upregulation of calprotectin in mild IgE-mediated ovalbumin hypersensitivity

doi: 10.18632/oncotarget.16954

Figure Lengend Snippet: Body weight ( A ) and length ( B ) were monitored weekly from the beginning of the study to the end. There was no significant difference between two groups from days 7 to days 42 (day 7 to 35, n = 5/group; day 42, n = 15/group, P all > 0.05).

Article Snippet: Immunohistochemistry was performed using standard protocols with the following antibodies: IL-1β, (1:100, both from Origene, Inc., Rockville, MD, USA); TNF-α, S100A8, S100A9 (1:200, Abcam, Inc., Cambridge, MA, USA); Toll-like receptor 4 (TLR4), S100A8/A9 (1:200, NOVUSbio, Inc., Littleton, CO, USA); nuclear factor kappa B (NF-κB) (1:200, Cell Signaling Technology, Inc., Danvers, MA, USA).

Techniques:

Journal: Oncotarget

Article Title: Upregulation of calprotectin in mild IgE-mediated ovalbumin hypersensitivity

doi: 10.18632/oncotarget.16954

Figure Lengend Snippet: PCR was used to detect the mRNA expression of TLR4, NF-κB, TNF-α, IL-1β, and IL-6. ( A , B , C , D , and E ) show the expression of TLR4, NF-κB, TNF-α, IL-1β, and IL-6 in jejunum; ( F , G , H , I and J ) show the expression of TLR4, NF-κB, TNF-α, IL-1β, and IL-6 mRNA in liver. * P < 0.05 ; # P < 0.01. ( K ) shows the expression of TLR4, NF-κB, TNF-α, and IL-1β. from left to right are control group, hypersensitivity groups from day 7 to day 42; and from top to bottom, expression of TLR4, NF-κB, TNF-α, and IL-1β in jejunum.

Article Snippet: Immunohistochemistry was performed using standard protocols with the following antibodies: IL-1β, (1:100, both from Origene, Inc., Rockville, MD, USA); TNF-α, S100A8, S100A9 (1:200, Abcam, Inc., Cambridge, MA, USA); Toll-like receptor 4 (TLR4), S100A8/A9 (1:200, NOVUSbio, Inc., Littleton, CO, USA); nuclear factor kappa B (NF-κB) (1:200, Cell Signaling Technology, Inc., Danvers, MA, USA).

Techniques: Expressing, Control