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: Cell & Bioscience

Article Title: The mechanisms by which hypothalamic neuroinflammation induced by neonatal cerebral ischemia–hypoxia leads to decreased thymic function via the HPA axis

doi: 10.1186/s13578-026-01543-w

Figure Lengend Snippet: HI induced the activations of TLR4/HMGB1/NF-κB signaling and microglia, leading to neuroinflammation in hypothalamus. A – B TNF-α and IL-1β on days 1, 3, 7, 14 and 21 after HI, n = 5–8. C Double immunofluorescence staining of Iba-1 and CD68. Scale bar = 20 μm. D Number of Iba-1 positive cells ( n = 4). E CD68 + / Iba-1 + cells rate ( n = 4). F – J CD11b, HMGB1, TLR4, p-P65-NF-κB and P65-NF-κB protein levels on day 7 after HI, n = 5. Data were expressed as the mean ± SD. Comparisons between the two groups were made using an unpaired T-test, * P < 0.05, ** P < 0.01, *** P < 0.001 versus Sham group.

Article Snippet: Serum hormone levels, along with hypothalamic tissue hormone and cytokine levels, were measured using CRH, ACTH, and CORT ELISA kits (Omnimabs, USA), as well as TNF-α and IL-1β ELISA kits (Proteintech, China), following the manufacturers’ instructions.

Techniques: Double Immunofluorescence Staining

Journal: Biology of Sex Differences

Article Title: Age and sex dependent effects of early overnutrition on metabolic parameters and the role of neonatal androgens

doi: 10.1186/s13293-016-0079-5

Figure Lengend Snippet: The limit of detection and intra- and inter-assay coefficients (CV) of variation for the hormone and cytokine assays

Article Snippet: Assay-on-demand kits (Applied Biosystems) were used to assess the mRNA levels of leptin (Rn00565158_m1), adiponectin (Rn00595250_m1), IL6 (Rn01410330_m1), IL1β (Rn01336189_m1), and TNFα (Rn01525859_g1) quantitative real-time PCR according to the manufacturer’s protocol and analyzed in an ABI PRISM 7000 Sequence Detection System (Applied Biosystems).

Techniques: Inter Assay, Intra Assay

Journal: Biology of Sex Differences

Article Title: Age and sex dependent effects of early overnutrition on metabolic parameters and the role of neonatal androgens

doi: 10.1186/s13293-016-0079-5

Figure Lengend Snippet: Glycemia and serum hormone and cytokine levels at postnatal days (PND) 10, 21, 30, 50, 85, and 150 in male (M) and female (F) rats raised in litters (L) 4 or 12 pups

Article Snippet: Assay-on-demand kits (Applied Biosystems) were used to assess the mRNA levels of leptin (Rn00565158_m1), adiponectin (Rn00595250_m1), IL6 (Rn01410330_m1), IL1β (Rn01336189_m1), and TNFα (Rn01525859_g1) quantitative real-time PCR according to the manufacturer’s protocol and analyzed in an ABI PRISM 7000 Sequence Detection System (Applied Biosystems).

Techniques:

Journal: Biology of Sex Differences

Article Title: Age and sex dependent effects of early overnutrition on metabolic parameters and the role of neonatal androgens

doi: 10.1186/s13293-016-0079-5

Figure Lengend Snippet: Relative mRNA levels of cytokines in inguinal and perigonadal adipose tissue at postnatal days (PND) 10, 21, 30, 50, 85, and 150 in male (M) and female (F) rats raised in litters (L) 4 or 12 pups

Article Snippet: Assay-on-demand kits (Applied Biosystems) were used to assess the mRNA levels of leptin (Rn00565158_m1), adiponectin (Rn00595250_m1), IL6 (Rn01410330_m1), IL1β (Rn01336189_m1), and TNFα (Rn01525859_g1) quantitative real-time PCR according to the manufacturer’s protocol and analyzed in an ABI PRISM 7000 Sequence Detection System (Applied Biosystems).

Techniques:

Journal: Biology of Sex Differences

Article Title: Age and sex dependent effects of early overnutrition on metabolic parameters and the role of neonatal androgens

doi: 10.1186/s13293-016-0079-5

Figure Lengend Snippet: Metabolic changes in males, females, and females that were androgenized (AF) on postnatal day (PND) 1. At PND10, there were significant differences in a the percentage of inguinal adipose tissue (IngAT) and serum b leptin, c adiponectin, and d interleukin (IL) 6 levels. At PND90, there were significant differences in e body weight, f glycemia and serum, g insulin, h leptin and i IL1β levels. The mRNA levels of j leptin, k adiponectin, l IL6, and m TNFα in perigonadal adipose tissue at PND90 were also affected. # p < 0.0001, ## p < 0.002, ### p < 0.0002, * p < 0.01, *** p < 0.005, @ p < 0.05. PND10: inguinal adipose tissue: N = 10; serum leptin and IL6 levels: N = 6; serum adiponectin: M and F: N = 8, AF: N = 9. PND90: body weight and glycemia: M: N = 8, F: N = 12, AF: N = 11; serum insulin, leptin, and Il1β levels: N = 6; mRNA levels for leptin, adiponectin, IL6, and TNFα: N = 6

Article Snippet: Assay-on-demand kits (Applied Biosystems) were used to assess the mRNA levels of leptin (Rn00565158_m1), adiponectin (Rn00595250_m1), IL6 (Rn01410330_m1), IL1β (Rn01336189_m1), and TNFα (Rn01525859_g1) quantitative real-time PCR according to the manufacturer’s protocol and analyzed in an ABI PRISM 7000 Sequence Detection System (Applied Biosystems).

Techniques:

Journal: BMC Molecular Biology

Article Title: A flow cytometry technique to study intracellular signals NF-κB and STAT3 in peripheral blood mononuclear cells

doi: 10.1186/1471-2199-8-64

Figure Lengend Snippet: Western blot and ELISA analysis of phosphorylated NF-κB and STAT3 in PBMCs . Western Blot of pNF-κB (A, C) and pSTAT3 (E) in PBMCs nuclear extract was observed with or without stimuli: IL1β (50 ng/mL) (A), sCD40L (50 ng/mL) (C) or IL10 (100 ng/mL) (E) for various periods (0 to 30 min). Data were representative of five experiments. pNF-κB and pSTAT3 were detected by ELISA (n = 5) in activated PBMCs nuclear extracts and the time dependant effect of the PBMCs stimulus was observed. PBMCs were stimulated for various periods of time (0 to 30 min) with or without IL1β (B), sCD40L (D) or IL10 (F). Statistical significance (wilcoxon paired test; p < 0.05) was represented by an asterisk (*). Data represented the mean (± SD) of five experiments.

Article Snippet: PBMCs were stimulated in the presence or absence of IL1β (AbCys, Paris, France), purified – soluble – trimeric human CD40L molecules (sCD40L) [ ] (Alexis-Coger, Paris, France) or IL10 (R&D System, Lille, France) at varying concentrations: IL1β (0 to 200 ng/mL), sCD40L (0 to 200 ng/mL) and IL10 (from 0 to 200 ng/mL) and for various time periods (0, 5, 15, 30 min).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay

Journal: BMC Molecular Biology

Article Title: A flow cytometry technique to study intracellular signals NF-κB and STAT3 in peripheral blood mononuclear cells

doi: 10.1186/1471-2199-8-64

Figure Lengend Snippet: Flow cytometry of simultaneous phosphorylated NF-κB and STAT3 expression from B-cells, T-lymphocytes and monocytes/macrophages stimulated by IL1β (control) . Cytogram (A and C) depicted one experiment and showed the isotype control (A1 and C1 for B-cells, A4 and C4 for T-lymphocytes and A7 and C7 for monocytes/macrophages). Cytogram (A) showed the pNF-κB translocation of B-cells (A2, A3), T-lymphocytes (A5, A6) and monocytes/macrophages (A8, A9) with (A3, A6, A9) or without (A2, A5, A8) stimulus IL1β (50 ng/mL) for 30 min. Cytogram (C) showed the pSTAT3 translocation of B-cells (C2, C3), T-lymphocytes (C5, C6) and monocytes/macrophages (C8, C9) with (C3, C6, C9) or without (C2, C5, C8) stimulus IL1β (50 ng/mL) for 30 min. Data were representative of seven experiments. Summary of flow cytometry analysis (n = 7) of percentage of phosphorylated NF-κB (B) and phosphorylated STAT3 (D) activation (versus untreated) from B-cells, T-lymphocytes and monocytes/macrophages. The graphs represent the difference in percentage of phosphorylated nuclear factor between stimulated and untreated cells. Statistical significance (wilcoxon paired test; p < 0.05) was represented by an asterisk (*). Data represented the mean (± SD) of seven experiments.

Article Snippet: PBMCs were stimulated in the presence or absence of IL1β (AbCys, Paris, France), purified – soluble – trimeric human CD40L molecules (sCD40L) [ ] (Alexis-Coger, Paris, France) or IL10 (R&D System, Lille, France) at varying concentrations: IL1β (0 to 200 ng/mL), sCD40L (0 to 200 ng/mL) and IL10 (from 0 to 200 ng/mL) and for various time periods (0, 5, 15, 30 min).

Techniques: Flow Cytometry, Expressing, Control, Translocation Assay, Activation Assay