Journal: Frontiers in Immunology

Article Title: Pathological neutrophil extracellular traps hinder postoperative anal fistula wound healing and are attenuated by Zuoqing granule via suppression of the Nox4 pathway

doi: 10.3389/fimmu.2025.1730184

Figure Lengend Snippet: ZQG attenuates PMA-induced NETosis and inflammatory responses in human neutrophils via targeting the Nox4/ROS pathway. (A) ZQG suppresses oxidative burst and NET formation. Representative immunofluorescence images showing intracellular ROS (stained by DHE, red) and NETosis (marked by CitH3, green). Nuclei are counterstained with DAPI (blue). Scale bars: 20 μm. (B) Quantitative analysis of intracellular ROS levels. (C) Quantification of NETosis, expressed as the percentage of Sytox Green + cells. (D) Western blot analysis demonstrating the effects of ZQG and si-Nox4 on the protein expression of Nox4, p-Akt, and PADI4. (E) Densitometric quantification of Western blot results. (F) qPCR analysis of Nox4, PI3K, Akt, and PADI4 mRNA expression. (G) ELISA analysis of pro-inflammatory cytokine levels (IL-2, IL-5, IL-6, IL-12, TNF-α) in cell culture supernatants. Data are presented as mean ± SEM (n=10 independent experiments). ***p < 0.001 vs. PMA group.

Article Snippet: The concentrations of interleukin-2 (IL-2), interleukin-5 (IL-5), interleukin-6 (IL-6), interleukin-12 (IL-12), and tumor necrosis factor-alpha (TNF-α) in the supernatants were quantified using specific commercial rat ELISA kits (R&D Systems, Catalog # R6000B) according to the manufacturers’ instructions.

Techniques: Immunofluorescence, Staining, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: JCI Insight

Article Title: Quinolinic acid potentially links kidney injury to brain toxicity

doi: 10.1172/jci.insight.180229

Figure Lengend Snippet: Kidney cortex RNA and protein lysates were prepared from Mdm2 -cKO vs. control mice ( n = 4 per group). ( A ) Transformation related protein 53 ( Trp53 ) mRNA and ( B ) p53 protein levels. ( C ) Cyclin-dependent kinase inhibitor 1A ( Cdkn1a ) mRNA and ( D ) p21 protein levels. ( E ) Cytochrome b-245, beta polypeptide ( Cybb ), mRNA and ( F ) CYBB protein levels. ( G ) CXC chemokine ligand 1 ( Cxcl1 ) mRNA level. ( H ) CXCL1 protein concentration. ( I ) Interleukin-1β ( Il1b ) mRNA level. ( J ) IL-1β protein concentration. ( K ) Neutrophil-to-lymphocyte ratio (VetScan Analyzer). ( L ) Plasma interleukin-6 (IL-6) protein concentration. ( M ) Plasma soluble urokinase plasminogen activator receptor (suPAR) concentration. Quantitative PCR (qPCR) results (normalized to Gapdh expression) and protein levels measured via Western blot (normalized to actin) are presented as fold-change relative to control. The proteins analyzed via ELISA are reported as concentrations. Graphs display means ± SEM. Two-tailed t tests: * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Quantitative protein levels in plasma and tissue lysates were assessed using ELISA kits following manufacturer’s protocols: mouse uPAR DuoSet ELISA (R&D Systems, Bio-Techne, DY531), mouse CXCL1/KC Quantikine ELISA Kit (R&D Systems, Bio-Techne, MKC00B-1), mouse IL-6 Quantikine ELISA Kit (R&D Systems, Bio-Techne, M6000B-1), and mouse IL-1β/IL-1F2 Quantikine ELISA Kit (R&D Systems, Bio-Techne, MLB00C-1).

Techniques: Control, Transformation Assay, Protein Concentration, Clinical Proteomics, Concentration Assay, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: JCI Insight

Article Title: Quinolinic acid potentially links kidney injury to brain toxicity

doi: 10.1172/jci.insight.180229

Figure Lengend Snippet: Mice were fed regular chow (control) or 0.2% adenine-enriched chow for 1 month ( n = 4 per group). ( A and B ) Plasma suPAR and IL-6 levels measured by ELISA. Concentrations of Trp, KYN, 3HK, QA, and 5-HIAA in ( C – G ) plasma, kidney cortex ( H – L ), and brain cortex ( M – Q ). QA and 5-HIAA were measured using a different mass spectrometry method. Graphs display means ± SEM. Two-tailed t test; * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Quantitative protein levels in plasma and tissue lysates were assessed using ELISA kits following manufacturer’s protocols: mouse uPAR DuoSet ELISA (R&D Systems, Bio-Techne, DY531), mouse CXCL1/KC Quantikine ELISA Kit (R&D Systems, Bio-Techne, MKC00B-1), mouse IL-6 Quantikine ELISA Kit (R&D Systems, Bio-Techne, M6000B-1), and mouse IL-1β/IL-1F2 Quantikine ELISA Kit (R&D Systems, Bio-Techne, MLB00C-1).

Techniques: Control, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Mass Spectrometry, Two Tailed Test