Journal: International Journal of Molecular Sciences

Article Title: miR-132-3p Modulates DUSP9-Dependent p38/JNK Signaling Pathways to Enhance Inflammation in the Amnion Leading to Labor

doi: 10.3390/ijms23031864

Figure Lengend Snippet: The role of p38 and JNK in DUSP9 siRNA-induced expression of proinflammatory cytokines and COX2 as well as PGE2 in WISH cells. WISH cells were transfected with si- DUSP9 or NC for 24 h, followed by 24 h of treatment with DMSO, p38 inhibitor SB203580 (10 μM), or JNK inhibitor SP600125 (20 μM). RT-qPCR analysis of the expression of IL-1β , TNF-α , COX2 ( A , G ), IL-6 , and IL-8 ( B , H ). Magnetic Luminex Assays of the secretion of IL-1β, TNF-α ( C , I ), IL-6, and IL-8 ( D , J ). Western blot analysis of COX2 expression ( E , K ). ELISA analysis of PGE2 level ( F , L ). Data were shown as mean ± SEM, n = 3. * p < 0.05, ** p < 0.01 vs. control (0), # p < 0.05, ## p < 0.01 vs. si- DUSP9 .

Article Snippet: To verify the involvement of p38 and JNK signaling pathways in the induction of IL-1β, IL-6, IL-8, TNF-α, and COX2 as well as PGE2 secretion by DUSP9 siRNA, the cells were transfected with NC or si- DUSP9 for 24 h, followed by 24 h of treatment with dimethyl sulfoxide (DMSO), p38 inhibitor SB203580 (10 μM, Selleck, Houston, TX, USA), or JNK inhibitor SP600125 (20 μM, Selleck, Houston, TX, USA).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Luminex, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: Frontiers in Pharmacology

Article Title: Establishment of a human nasal epithelium model of histamine-induced inflammation to assess the activity of fexofenadine as an inverse agonist and its link to clinical benefit

doi: 10.3389/fphar.2024.1393702

Figure Lengend Snippet: Reconstituted human nasal epithelium model (MucilAir™) to study the effect of HIS and FEX. CBF, cilia beating frequency; ELISA, enzyme-linked immunosorbent assay; LDH, lactate dehydrogenase; MCC, mucociliary clearance; RT-qPCR, quantitative reverse transcription–polymerase chain reaction; TEER, trans-epithelial electrical resistance.

Article Snippet: The basolateral medium was collected and measured using a Cytotoxicity Lactate Dehydrogenase (LDH) Assay Kit, following the manufacturer’s instructions (WST Dojindo, CK12-20).

Techniques: Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Reverse Transcription, Polymerase Chain Reaction

Journal: Cell reports

Article Title: Nanoparticle-based itaconate treatment recapitulates low-cholesterol/low-fat diet-induced atherosclerotic plaque resolution

doi: 10.1016/j.celrep.2024.114911

Figure Lengend Snippet:

Article Snippet: Direct-zol RNA Miniprep Kit , Zymo Research , R2054.

Techniques: Purification, Plasmid Preparation, Produced, Recombinant, Concentration Assay, Saline, Labeling, Membrane, Enzyme-linked Immunosorbent Assay, Quantitation Assay, Phospholipid Assay, BIA-KA, RNAscope, HD Assay, Polymer, Sequencing, Expressing, Software, Microscopy

Journal: European Journal of Medical Research

Article Title: Olfactory mucosa-mesenchymal stem cells with overexpressed Nrf2 modulate angiogenesis and exert anti-inflammation effect in an in vitro traumatic brain injury model

doi: 10.1186/s40001-025-02344-6

Figure Lengend Snippet: Effects of OM-MSCs Nrf2 on the levels of relevant cytokines in PC12 cells. A Flow cytometry was used to characterize CD45, CD34, CD29, CD90, CD73, and CD105 of OM-MSCs. B RT-qPCR-based to assess transfection efficiency after overexpression of Nrf2 plasmid. OM-MSCs Nrf2 : NRF2 overexpression group. OM-MSCs (ctrl): Empty vector transfection group. OM-MSCs: Untransfected OM-MSCs. C Morphology of both OM-MSCs (ctrl) and OM-MSCs Nrf2 under the optical microscope. Scale bar: 100 μm. D The changes in cytokine concentrations in PC12 cells measured by ELISA, including IL4, IL10, IL13, VEGF-A, and TGF-β. Results of independent triplicates were expressed as mean ± standard deviation and the statistical difference between two groups was marked with asterisks (“ns” indicates no significant difference, * p < 0.05, and ** p < 0.01). OM-MSCs olfactory mucosa-mesenchymal stem cells, Nrf2 : Nuclear Factor Erythroid-Derived 2-Like 2, IL interleukin, VEGF-A vascular endothelial growth factor-A, TGF-β transforming growth factor beta

Article Snippet: Specifically, PC12 cells were cultured in Roswell Park Memorial Institute-1640 medium (#C3250, Solarbio® Life Sciences, China) supplemented with 15% horse serum (#S9050, Solarbio® Life Sciences, China) and 5% fetal bovine serum; BV-2 cells were cultured in high-glucose DMEM (#D5194, Solarbio® Life Sciences, China) supplemented with 10% fetal bovine serum; HUVECs were maintained in M199 medium (#11150067, Gibco, USA) including 15% fetal bovine serum, 1% penicillin–streptomycin, 30 μg/mL endothelial cell growth supplement (#211-GS, Sigma, Darmstadt, Germany), and 10 ng/mL epidermal growth factor (#E5036, Sigma, Germany).

Techniques: Flow Cytometry, Quantitative RT-PCR, Transfection, Over Expression, Plasmid Preparation, Microscopy, Enzyme-linked Immunosorbent Assay, Standard Deviation, Derivative Assay

Journal: European Journal of Medical Research

Article Title: Olfactory mucosa-mesenchymal stem cells with overexpressed Nrf2 modulate angiogenesis and exert anti-inflammation effect in an in vitro traumatic brain injury model

doi: 10.1186/s40001-025-02344-6

Figure Lengend Snippet: Effects of OM-MSCs Nrf2 on the survival and angiogenesis and proteins related to PI3K/AKT pathway and apoptosis in PC12 cells. A Effects of OM-MSCs (ctrl) and OM-MSCs Nrf2 on the survival of PC12 cells based on CCK-8 assay. B , C Angiogenesis assay in evaluating the angiogenesis of PC12 cells following the co-culture with OM-MSCs (ctrl) and OM-MSCs Nrf2 . scale bar: 200 μm D , E Ratio of p-PI3K to PI3K and p-AKT to AKT as well as BAX protein level in PC12 cells following the co-culture with OM-MSCs (ctrl) and OM-MSCs Nrf2 , and those cells treated with minocycline were applied as the positive control. Results of independent triplicates were expressed as mean ± standard deviation and the statistical difference between two groups was marked with asterisks ( * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001). OM-MSCs olfactory mucosa-mesenchymal stem cells, Nrf2 Nuclear Factor Erythroid-Derived 2-Like 2, OD optical density, p-PI3K phosphorylated-PI3K, PI3K Phosphoinositide 3-kinase, p-AKT phosphorylated-AKT, AKT protein kinase B, BAX BCL2 Associated X, GAPDH Glyceraldehyde-3-phosphate dehydrogenase

Article Snippet: Specifically, PC12 cells were cultured in Roswell Park Memorial Institute-1640 medium (#C3250, Solarbio® Life Sciences, China) supplemented with 15% horse serum (#S9050, Solarbio® Life Sciences, China) and 5% fetal bovine serum; BV-2 cells were cultured in high-glucose DMEM (#D5194, Solarbio® Life Sciences, China) supplemented with 10% fetal bovine serum; HUVECs were maintained in M199 medium (#11150067, Gibco, USA) including 15% fetal bovine serum, 1% penicillin–streptomycin, 30 μg/mL endothelial cell growth supplement (#211-GS, Sigma, Darmstadt, Germany), and 10 ng/mL epidermal growth factor (#E5036, Sigma, Germany).

Techniques: CCK-8 Assay, Angiogenesis Assay, Co-Culture Assay, Positive Control, Standard Deviation, Derivative Assay

Journal: European Journal of Medical Research

Article Title: Olfactory mucosa-mesenchymal stem cells with overexpressed Nrf2 modulate angiogenesis and exert anti-inflammation effect in an in vitro traumatic brain injury model

doi: 10.1186/s40001-025-02344-6

Figure Lengend Snippet: Relevant mechanisms related to OM-MSCs Nrf2 on the neuroinflammation mediators and glycolysis in PC12 cells. A Effects of OM-MSCs and OM-MSCs Nrf2 on the mRNA level of Nlrp3 , Il1b , Il18 , Ccl5 , P2ry12 , and Cx3cr1 in PC12 cells. B , C Effects of OM-MSCs (ctrl) and OM-MSCs Nrf2 on the ECAR ( B ) and OCR ( C ) of PC12 cells. Results of independent triplicates were expressed as mean ± standard deviation and the statistical difference between two groups was marked with asterisks ( * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001). OM-MSCs olfactory mucosa-mesenchymal stem cells, Nrf2 Nuclear Factor Erythroid-Derived 2-Like 2, Nlrp3 NOD-, LRR- and pyrin domain-containing protein 3; Il1b Interleukin 1 beta; Il18 Interleukin 18, Ccl5 C–C chemokine ligand 5, P2ry12 purinergic receptor P2Y12, Cx3cr1 C-X3-C motif chemokine receptor 1, ECAR extracellular acidification rate, oligo Oligomycin; OCR oxygen consumption rate, 2-DG 2-deoxy-D-glucose, FCCP Trifluoromethoxy carbonylcyanide phenylhydrazone

Article Snippet: Specifically, PC12 cells were cultured in Roswell Park Memorial Institute-1640 medium (#C3250, Solarbio® Life Sciences, China) supplemented with 15% horse serum (#S9050, Solarbio® Life Sciences, China) and 5% fetal bovine serum; BV-2 cells were cultured in high-glucose DMEM (#D5194, Solarbio® Life Sciences, China) supplemented with 10% fetal bovine serum; HUVECs were maintained in M199 medium (#11150067, Gibco, USA) including 15% fetal bovine serum, 1% penicillin–streptomycin, 30 μg/mL endothelial cell growth supplement (#211-GS, Sigma, Darmstadt, Germany), and 10 ng/mL epidermal growth factor (#E5036, Sigma, Germany).

Techniques: Standard Deviation, Derivative Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Neutrophil-macrophage crosstalk via NETs–IL-17/VEGF/S100A9 axis promotes hepatocellular carcinoma progression

doi: 10.1186/s13046-025-03618-x

Figure Lengend Snippet: The levels of NETs increase in HCC patients, especially in advanced and metastatic stages. A A heatmap illustrating the differential expression patterns of NETs-related genes in normal and HCC samples based on TCGA database analysis. B Box plots depicting the expression differences of NETs-related genes between normal and HCC samples from the TCGA database. C , D Representative images of HE (Hematoxylin and eosin) and IF staining for CD66b (C) and CitH3 (D) in tissue sections from HCs and HCC patients. The mean IOD of CD66b or CitH3 was quantified (Image Pro Plus) from three random regions per section (HC, n = 5; HCC, n = 5) and shown as fold change relative to HCs. E Representative images of IF staining for CitH3 in peripheral blood neutrophils from HC and HCC patient F Western blot analysis of CitH3 expression in peripheral blood neutrophils from HCs ( n = 8) and HCC ( n = 8) patients, with band intensities normalized to β-actin and shown as fold change relative to controls G ELISA analysis of serum MPO-DNA levels in HCs and HCC patients. H ELISA analysis of serum MPO-DNA levels in HCC patients at different TNM stages (I–IV). I ELISA analysis of serum MPO-DNA levels in HCC patients with and without metastasis. J ELISA analysis of serum MPO-DNA levels in HCC patients with intrahepatic metastasis and extrahepatic metastasis. K Western blot analysis of CitH3 expression in peripheral blood neutrophils from HCC patients with ( n = 6) and without ( n = 6) extrahepatic metastasis. Band intensities were normalized to β-actin and quantified relative to the non-metastasis group White and black scale bars: 50 μm. Data are presented as median (IQR) (G-J) and other data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. Student’s t test (C, D, F, K), Mann–Whitney U test (B, G, I, J), Kruskal–Wallis test (H)

Article Snippet: Neutrophils were isolated from the peripheral blood of healthy volunteers and HCC patients using the Human Peripheral Blood Neutrophil Isolation Kit (P9040, Solarbio, China), according to the manufacturer’s instructions.

Techniques: Quantitative Proteomics, Expressing, Staining, Western Blot, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Neutrophil-macrophage crosstalk via NETs–IL-17/VEGF/S100A9 axis promotes hepatocellular carcinoma progression

doi: 10.1186/s13046-025-03618-x

Figure Lengend Snippet: Activation of IL-17R/NF-κB signaling by NETs-attached IL-17 is responsible for NETs-induced M2 macrophage polarization. A Schematic illustration of the transcriptome sequencing protocol for THP1-M0 under three treatment conditions. The volcano plot shows significantly DEGs for Group B vs. A, Group C vs. A, and Group C vs. B. B KEGG pathway enrichment analysis of DEGs was conducted using the DAVID tool. C FCM analysis of the proportion of CD68 + CD206 + M2 derived from THP-1-M0 following 48-hour co-culture of HuH-7 cells with NETs and (or) AMG827. Quantification of the fold change in the proportion from three independent experiments is shown in the right panel. D Western blot analysis of IL-10 and VEGF expression in cell lysates following the same treatments as in C. Densitometric values were normalized to β-actin and compared to the control group, with quantification shown in the right panel E Western blot analysis of IL-17 expression in extracted and purified NETs from peripheral blood neutrophils of HCs ( n = 8) and HCC patients ( n = 8). F ELISA analysis of IL-17 levels carried by NETs standardized to 50 ng/µL in HCs ( n = 8) and patients with HCC ( n = 8). G Western blot analysis of IL-17 expression in peripheral blood neutrophils from HCs ( n = 8) and HCC ( n = 8) patients. Densitometric values were normalized to β-actin and shown as a fold change relative to HCs. H Representative IF images of IL-17 and NE staining in peripheral blood neutrophils with (lower panel) or without PMA stimulation (upper panel) for 4 h from HCs and HCC patients. I Western blot analysis was conducted to assess the expression levels of p-65 and p-p65(S536) in THP-1-M0 following 48 h co-culture of HuH-7 cells with NETs and (or) AMG827. Densitometric values were normalized to β-actin and shown as fold changes relative to control in the right panel. J FCM was used to analyze the proportion of CD68 + CD206 + M2 derived from THP-1-M0 following 48-hour co-culture of HuH-7 cells with NETs and (or) BAY11-7082. Quantification of the fold change in the proportion from three independent experiments is shown in the right panel. K IF analysis of CD206 + M2 macrophage from the above co-culture systems of J treatment for 48 h. L ELISA analysis of secretion levels of M2d-related cytokines VEGF, IL-10 and TGF-β, in the culture supernatant from the above co-culture systems of J treatment for 48 h White scale bars: 20 μm. Data are presented as mean ± SD. Ns, not significant. * p < 0.05, ** p < 0.01, *** p < 0.001. One-way ANOVA followed by the Newman-Keuls multiple comparison test (C, D, I, J, L); Student’s t test (F, G)

Article Snippet: Neutrophils were isolated from the peripheral blood of healthy volunteers and HCC patients using the Human Peripheral Blood Neutrophil Isolation Kit (P9040, Solarbio, China), according to the manufacturer’s instructions.

Techniques: Activation Assay, Sequencing, Derivative Assay, Co-Culture Assay, Western Blot, Expressing, Control, Purification, Enzyme-linked Immunosorbent Assay, Staining, Comparison