Journal: PLoS ONE

Article Title: Neutrophils remain detrimentally active in hydroxyurea-treated patients with sickle cell disease

doi: 10.1371/journal.pone.0226583

Figure Lengend Snippet: (A) Purified and rested neutrophils from healthy donors (H) and SCD patients in steady state (SS) were left untreated in RPMI as control or treated with 20 μM hemin. Median Fluorescence Intensity of F-Actin (upper panel) and CD63 (lower panel) in F-Actin/CD63 double positive neutrophils with normal multi-lobulated nuclei is shown. (Healthy N = 7; SCD N = 6). (B) Z-stack stills of SCD neutrophils left in RPMI for 30 minutes showing neutrophils with roughed cell membranes (yellow arrows, upper panel). Staining for DNA (blue), F-Actin (green) and CD63 (red) is presented (lower panel). (C) MPO release was assayed by ELISA following 30 minutes incubation with or without the hemin stimulus (H (Healthy) N = 4; SS (SCD) N = 4). Degranulation response was assayed in terms of MPO ng/ml plasma (upper panel), or fold increase over healthy neutrophils (lower panel). Data presented as dot plots±S.D., significance calculated with an unpaired Mann-Whitney test.

Article Snippet: The supernatants were then assayed with a Human Myeloperoxidase Quantikine ELISA Kit from R&D Systems according to the manufacturer’s instruction.

Techniques: Purification, Control, Fluorescence, Staining, Enzyme-linked Immunosorbent Assay, Incubation, Clinical Proteomics, MANN-WHITNEY

Journal: Theranostics

Article Title: Blockade of phosphotyrosine pathways suggesting SH2 superbinder as a novel therapy for pulmonary fibrosis

doi: 10.7150/thno.72269

Figure Lengend Snippet: SH2 superbinder exhibited excellent security by targeting lung fibroblasts based on pY levels in vivo. A , Schematic showing progression of SH2 superbinder treatment in control mice. B , Percent survival during last 7 days after SH2 superbinder injection (n = 8). C , HE staining after SH2 superbinder challenge. Images showing the panoramic and partial view of lungs. D , Neutrophil accumulation measured by MPO assay in lung tissues after GST, GST-SH2 WT, GST-SH2 TrM or SH2-TrM treatment in saline groups (n = 6). E , Total cell counting in BALF (n = 6). F-G , Changes in different kinds of cells by Giemsa and Diff-quick staining in BALF (n = 6). H , Inflammatory cytokines in BALF measured by ELISA (n = 6). I , The determination of liver function index in serum (n = 6). J , Schematic showing progression of SH2 superbinder treatment in BLM-treated mice. K , Western blot of GST tag of different tissues in BLM group treated with GST-SH2 TrM. L , The process of FACS for mice lungs. M , Western blot of pY levels in different lung cells which were isolated by FACS of BLM treated 14 days mice. N , The fluorescence images of SH2 superbinder entering into activated fibroblasts, epithelial cells (EpCAM + ), leukocytes (CD45 + ) and endothelial cells (CD31 + ). O-R , Representative images of SH2 superbinder in myofibroblast (α-SMA + ) ( O ), epithelial cells (EpCAM + ) ( P ), leukocytes (CD45 + ) ( Q ) and endothelial cells (CD31 + ) ( R ) of BLM-treated mice.

Article Snippet: The amounts of IL-1β, IL-6, IL-10 and TNF-α in BALF of mice were measured by using mouse ELISA kits from R&D Systems (Minneapolis, MN, USA).

Techniques: In Vivo, Control, Injection, Staining, MPO Assay, Saline, Cell Counting, Diff-Quik, Enzyme-linked Immunosorbent Assay, Western Blot, Isolation, Fluorescence

Journal: Biomarker Research

Article Title: ZDHHC5-mediated S-palmitoylation of NOD2 drives neutrophil extracellular trap formation via mitophagy to exacerbate inflammation in COPD

doi: 10.1186/s40364-026-01001-2

Figure Lengend Snippet: NOD2 deficiency ameliorates cigarette smoke-induced lung inflammation and injury in vivo The functional role of NOD2 was evaluated in a cigarette smoke (CS)-induced mouse model of COPD using wild-type (wt) and NOD2-knockout (NOD2-/-) mice, with lentiviral-mediated NOD2 re-expression (NOD2-OE) in NOD2-/- COPD mice. ( A ) Schematic of the in vivo experimental design. ( B ) qRT-PCR analysis of NOD2 mRNA expression in lung tissue. ( C ) Representative images of hematoxylin and eosin (H&E) staining of lung tissue sections. Scale bars: 200 μm in the upper panels and 100 μm in the enlarged lower panels. ( D ) Representative images of Masson’s trichrome staining for collagen deposition in lung tissue sections. Scale bar: 100 μm. ( E ) Quantification of total cells, neutrophils, and macrophages in bronchoalveolar lavage fluid (BALF), and ELISA measurements of TNF-α, IL-6, and CXCL1 levels in BALF supernatant. ( F ) Representative images of IHC staining for E-cadherin and N-cadherin in lung tissue. Scale bar: 50 μm. ( G ) Quantification of the average optical density (AOD) for E-cadherin and N-cadherin IHC staining. ( H ) qRT-PCR analysis of senescence-associated secretory phenotype (SASP) factors (CSF3, CCL3, and IL1α) in lung tissue. Data are presented as mean ± SD. Data were analyzed using one-way ANOVA followed by Tukey’s test. * p < 0.05 and ** p < 0.01 Control vs. wt-COPD groups; # p < 0.05 and ## p < 0.01 wt-COPD vs. NOD2 −/− -COPD groups; & p < 0.05 and && p < 0.01 NOD2 −/− -COPD + vector vs. NOD2 −/− -COPD + NOD2-OE groups

Article Snippet: Additionally, the commercial ELISA kits (Human and mouse MPO ELISA Kits, Cat. # CSB-E08721h, CSB-E08723m, CUSABIO, Wuhan, China; Human and mouse neutrophil elastase ELISA Kit, Cat. # CSB-EL007587HU, CSB-E04804m, CUSABIO) were applied to quantify the levels of MPO and neutrophil elastase released into cell culture supernatants as per the instructions of the manufacturer.

Techniques: In Vivo, Functional Assay, Knock-Out, Expressing, Quantitative RT-PCR, Analysis, Tissue, Staining, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Control, Plasmid Preparation

Journal: Biomarker Research

Article Title: ZDHHC5-mediated S-palmitoylation of NOD2 drives neutrophil extracellular trap formation via mitophagy to exacerbate inflammation in COPD

doi: 10.1186/s40364-026-01001-2

Figure Lengend Snippet: NOD2 promotes Neutrophil Extracellular Trap (NET) formation in the CS-induced COPD mouse model The association between NOD2, autophagy, and Neutrophil Extracellular Trap (NET) formation was investigated in the lung tissue of mice from the CS-induced COPD model. ( A) Representative immunofluorescence images showing co-localization of the neutrophil marker Ly6G (green) and the autophagy marker LC3B (red), with nuclei counterstained with DAPI (blue). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. ( B ) Representative immunofluorescence images showing staining for the NET markers citrullinated histone H3 (CitH3, green) and myeloperoxidase (MPO, red), with nuclei counterstained with DAPI (blue). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. ( C ) Quantification of the NET biomarkers CitH3-DNA and MPO-DNA complexes in serum and lung tissue homogenates by ELISA. ( D ) Immunoblotting analysis of the neutrophil marker Ly6G and the NET marker CitH3 in whole-lung tissue lysates. ( E ) Representative immunofluorescence images showing colocalization of the neutrophil marker Ly6G (red), the NET marker CitH3 (green), and NOD2 (pink) in lung tissue, with nuclei counterstained with DAPI (blue). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. Data are presented as mean ± SD. C and D were analyzed using one-way ANOVA followed by Tukey’s test. ** p < 0.01 Control vs. wt -COPD groups; and ## p < 0.01 wt -COPD vs. NOD2 −/− -COPD groups; && p < 0.01 NOD2 −/− -COPD + vector vs. NOD2 −/− -COPD + NOD2-OE groups

Article Snippet: Additionally, the commercial ELISA kits (Human and mouse MPO ELISA Kits, Cat. # CSB-E08721h, CSB-E08723m, CUSABIO, Wuhan, China; Human and mouse neutrophil elastase ELISA Kit, Cat. # CSB-EL007587HU, CSB-E04804m, CUSABIO) were applied to quantify the levels of MPO and neutrophil elastase released into cell culture supernatants as per the instructions of the manufacturer.

Techniques: Tissue, Immunofluorescence, Marker, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Analysis, Control, Plasmid Preparation

Journal: Biomarker Research

Article Title: ZDHHC5-mediated S-palmitoylation of NOD2 drives neutrophil extracellular trap formation via mitophagy to exacerbate inflammation in COPD

doi: 10.1186/s40364-026-01001-2

Figure Lengend Snippet: Neutrophil depletion minimizes genotype-dependent differences in CS-induced lung injury, inflammation, fibrosis, and NET formation The dependence of NOD2-associated COPD pathology on neutrophils was evaluated by depleting neutrophils in CS-exposed wild-type (wt) and NOD2 −/− mice. Mice were randomly assigned to the wt -COPD + IgG, wt -COPD + Anti-Ly6G, NOD2 −/− -COPD + IgG, and NOD2 −/− -COPD + Anti-Ly6G groups ( n = 6 per group). ( A ) Schematic of the experimental design. Mice were exposed to CS for 12 weeks and received 200 µg of anti-Ly6G antibody or an equivalent dose of isotype IgG intraperitoneally three times per week from week 2 until the end of the experiment. ( B ) Representative flow-cytometry plots and quantification of CD11b + Ly6G + neutrophils in lung tissue and peripheral blood. ( C ) Representative H&E-stained images of lung tissue sections. Scale bars: 200 μm in the upper panels and 100 μm in the enlarged lower panels. ( D ) Representative images of Masson’s trichrome staining for pulmonary collagen deposition. Scale bars: 100 μm. ( E ) Quantification of total cells, neutrophils, and macrophages in bronchoalveolar lavage fluid (BALF), together with ELISA measurements of TNF-α, IL-6, and CXCL1 levels in BALF supernatants. ( F ) Quantification of CitH3-DNA and MPO-DNA complexes in serum and lung tissue homogenates by ELISA. Data are presented as mean ± SD. Data were analyzed using two-way ANOVA followed by Tukey’s test. ** p < 0.01 vs. the wt -COPD + IgG group; ## p < 0.01 vs. the wt-COPD + Anti-Ly6G group; & p < 0.05, & p < 0.01 vs. the NOD2 −/− -COPD + IgG group

Article Snippet: Additionally, the commercial ELISA kits (Human and mouse MPO ELISA Kits, Cat. # CSB-E08721h, CSB-E08723m, CUSABIO, Wuhan, China; Human and mouse neutrophil elastase ELISA Kit, Cat. # CSB-EL007587HU, CSB-E04804m, CUSABIO) were applied to quantify the levels of MPO and neutrophil elastase released into cell culture supernatants as per the instructions of the manufacturer.

Techniques: Pathology, Flow Cytometry, Tissue, Staining, Enzyme-linked Immunosorbent Assay

Journal: Biomarker Research

Article Title: ZDHHC5-mediated S-palmitoylation of NOD2 drives neutrophil extracellular trap formation via mitophagy to exacerbate inflammation in COPD

doi: 10.1186/s40364-026-01001-2

Figure Lengend Snippet: Knockdown of NOD2 inhibits CSE-induced NET formation, ROS production, and mitophagy in human neutrophils The role of NOD2 in cigarette smoke extract (CSE)-induced Neutrophil Extracellular Trap (NET) formation, reactive oxygen species (ROS) accumulation, and lysosome–mitochondrion spatial relationships were evaluated in differentiated HL-60-derived neutrophil-like cells. ( A ) Schematic of the in vitro experimental design and treatment groups. ( B ) Representative DAPI-stained images of extracellular DNA structures and quantification of NET formation based on DAPI-stained nuclear morphology. Scale bars: 20 μm. ( C ) Extracellular DNA levels in cell culture supernatants determined using a PicoGreen dsDNA assay. ( D ) Myeloperoxidase (MPO) levels in cell culture supernatants determined by ELISA. ( E ) Neutrophil elastase (NE) levels in cell culture supernatants determined by ELISA. ( F ) Representative flow-cytometry histograms and quantitative analysis of intracellular ROS levels measured using DCFH-DA staining. ( G ) Representative immunofluorescence images showing the spatial relationship between LAMP1-positive lysosomes (green) and TOM20-positive mitochondria (red), with nuclei counterstained with DAPI (blue). The colocation efficients for LAMP1 and TOM20 was determined by ImageJ and shown on the right. Scale bars: 20 μm. Data are presented as mean ± SD. Quantitative data were analyzed using one-way ANOVA followed by Tukey’s test. ** p < 0.01 vs. Control group; ## p < 0.01 vs. CSE + sh-NC group

Article Snippet: Additionally, the commercial ELISA kits (Human and mouse MPO ELISA Kits, Cat. # CSB-E08721h, CSB-E08723m, CUSABIO, Wuhan, China; Human and mouse neutrophil elastase ELISA Kit, Cat. # CSB-EL007587HU, CSB-E04804m, CUSABIO) were applied to quantify the levels of MPO and neutrophil elastase released into cell culture supernatants as per the instructions of the manufacturer.

Techniques: Knockdown, Derivative Assay, In Vitro, Staining, Morphology, Cell Culture, Picogreen Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Analysis, Immunofluorescence, Control

Journal: Biomarker Research

Article Title: ZDHHC5-mediated S-palmitoylation of NOD2 drives neutrophil extracellular trap formation via mitophagy to exacerbate inflammation in COPD

doi: 10.1186/s40364-026-01001-2

Figure Lengend Snippet: ZDHHC5 overexpression partially counteracts the effects of NOD2 knockdown in human neutrophil-like cells The ability of ZDHHC5 overexpression to counteract the effects of NOD2 knockdown was evaluated in CSE-stimulated, differentiated HL-60-derived neutrophil-like cells. ( A ) Schematic of the in vitro experimental design and treatment groups. ( B ) Representative DAPI-stained images of extracellular DNA structures and quantification of NET formation based on DAPI-stained nuclear morphology. Scale bars: 20 μm. ( C ) Extracellular DNA levels in cell culture supernatants determined using a PicoGreen dsDNA assay. ( D ) MPO levels in cell culture supernatants determined by ELISA. ( E ) NE levels in cell culture supernatants determined by ELISA. ( F ) Representative flow cytometry histograms and quantitative analysis of intracellular ROS levels measured using DCFH-DA staining. ( G ) Representative immunofluorescence images showing LAMP1-positive lysosomes (green) and TOM20-positive mitochondria (red), with nuclei counterstained with DAPI (blue). The colocalization coefficients for LAMP1 and TOM20 were determined by ImageJ and shown on the right. Scale bars: 20 μm. Data are presented as mean ± SD. Data were analyzed using two-way ANOVA followed Tukey’s test. ** p < 0.01 versus the CSE + sh-NC + Vector group; ## p < 0.01 versus the CSE + sh-NOD2 + ZDHHC5 group

Article Snippet: Additionally, the commercial ELISA kits (Human and mouse MPO ELISA Kits, Cat. # CSB-E08721h, CSB-E08723m, CUSABIO, Wuhan, China; Human and mouse neutrophil elastase ELISA Kit, Cat. # CSB-EL007587HU, CSB-E04804m, CUSABIO) were applied to quantify the levels of MPO and neutrophil elastase released into cell culture supernatants as per the instructions of the manufacturer.

Techniques: Over Expression, Knockdown, Derivative Assay, In Vitro, Staining, Morphology, Cell Culture, Picogreen Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Analysis, Immunofluorescence, Plasmid Preparation

Journal: Biomarker Research

Article Title: ZDHHC5-mediated S-palmitoylation of NOD2 drives neutrophil extracellular trap formation via mitophagy to exacerbate inflammation in COPD

doi: 10.1186/s40364-026-01001-2

Figure Lengend Snippet: ZDHHC5 overexpression partially counteracts the protective effects of NOD2 knockdown in a mouse model of COPD The ability of ZDHHC5 overexpression to counteract the effects of NOD2 knockdown was evaluated in a cigarette smoke (CS)-induced mouse model of COPD. ( A ) Schematic of the in vivo experimental design. ( B ) qRT-PCR analysis of NOD2 and ZDHHC5 mRNA expression in lung tissue. ( C ) Representative images of H&E staining of lung sections. Scale bar: 100 μm. ( D ) Representative images of Masson’s trichrome staining for collagen deposition. Scale bar: 100 μm. ( E ) Quantification of inflammatory cells and TNF-α, IL-6, and CXCL1 levels in BALF. ( F ) Representative images of IHC staining for E-cadherin and N-cadherin. Scale bar: 50 μm. ( G ) Quantitative analysis of IHC staining. ( H ) Immunofluorescence images showing spatial overlap between Ly6G (green) and LC3B (red). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. ( I ) Immunofluorescence images showing NETs stained for CitH3 (green) and MPO (red). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. ( J ) Quantification of CitH3-DNA and MPO-DNA complexes in serum and lung tissue homogenates by ELISA. Data are presented as mean ± SD. Quantitative data were analyzed using two-way ANOVA followed by Tukey’s test. * p < 0.05, ** p < 0.01 vs. COPD + sh-NC + Vector group; # p < 0.05, ## p < 0.01 vs. COPD + sh-NOD2 + ZDHHC5 group

Article Snippet: Additionally, the commercial ELISA kits (Human and mouse MPO ELISA Kits, Cat. # CSB-E08721h, CSB-E08723m, CUSABIO, Wuhan, China; Human and mouse neutrophil elastase ELISA Kit, Cat. # CSB-EL007587HU, CSB-E04804m, CUSABIO) were applied to quantify the levels of MPO and neutrophil elastase released into cell culture supernatants as per the instructions of the manufacturer.

Techniques: Over Expression, Knockdown, In Vivo, Quantitative RT-PCR, Analysis, Expressing, Tissue, Staining, Immunohistochemistry, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Plasmid Preparation

Journal: Stroke

Article Title: Phosphatidylinositol-3-Kinase Gamma Plays a Central Role in Blood–Brain Barrier Dysfunction in Acute Experimental Stroke

doi: 10.1161/strokeaha.110.601369

Figure Lengend Snippet: Figure 5. PI3K deficiency reduces neutrophil infiltration and neutrophil-associated MMP-9. A–B, Immunostaining for neutrophils (A) and MMP-9 (B) in the ischemic cortex in WT and PI3K KO mice at the indicated times after tMCAO. The number of the cells positively stained for neutrophils and MMP-9 was calculated in the 3 predefined regions of interest shown in adjoining coronal section. Bar50 m. n5 per time point per group; *P0.05 vs WT control. C, Neutrophil infiltration was quantified by myeloperoxidase (MPO) assay. Enzyme-linked immunosorbent assay (ELISA) assay of MPO protein in the ischemic cortex (region of interest shown in adjoining coronal section) 24 hours after tMCAO. n5 for each group. *P0.05 vs WT control. In A–C, samples from sham-operated animals served as controls (Ctrl). D, Double immunostaining showing the colocalization of MMP-9 with the specific neutrophil marker (PMN), but not with microglial (Iba-1), astrocytic (GFAP), or neuronal (NeuN) markers in the ischemic cortex at 24 hours after tMCAO. Bar50 m. PI3K indicates phosphatidylinositol-3- kinase-gamma; MMP-9, metallopeptidase-9; WT, wild-type; KO, knockout; tMCAO, transient occlusion of the middle cerebral artery; Iba-1, ionized calcium-binding adapter molecule 1; GFAP, glial fibrillary acidic protein; NeuN, neuronal nuclei.

Article Snippet: Enzyme-Linked Immunosorbent Assays Enzyme-linked immunosorbent assays of brain cortice homogenates were performed to detect total MMP-9 protein using a Quantikine mouse MMP-9 (total) immunoassay kit (R&D Systems), myeloperoxidase using the mouse Enzyme-linked immunosorbent assay kit (HK210; Hycult Biotechnology), and malondialdehyde (MDA) using the NWLSS MDA assay kit (Northwest Life Science Specialties, LLC) according to the instructions by the manufacturers.

Techniques: Immunostaining, Staining, Control, MPO Assay, Enzyme-linked Immunosorbent Assay, Double Immunostaining, Marker, Knock-Out, Binding Assay

Journal: Stroke

Article Title: Phosphatidylinositol-3-Kinase Gamma Plays a Central Role in Blood–Brain Barrier Dysfunction in Acute Experimental Stroke

doi: 10.1161/strokeaha.110.601369

Figure Lengend Snippet: Figure 6. PI3K deficiency reduces MMP-9 activity and protein content. A, Gelatin zymogram. Note that MMP-9 (92 and 82 kDa) and MMP-2 (63 kDa) bands were detected in the homogenates of ischemic (left [L]) and contralateral (right [R]) cortex 24 hours after tMCAO. The region of interest is shown in adjoining coronal section. B, Densitometric analysis of the bands shown in A. Data represent meanSD from 3 separate experiments. *P0.05 vs WT (saline-treated) control. C, Enzyme-linked immunosorbent assay (ELISA) assay of MMP-9 protein in ischemic (left) and contralateral (right) cortex (region of interest shown in adjoining coronal section) 24 hours after tMCAO. n5 for each group. **P0.01 vs WT control. PI3K indicates phosphatidylinositol-3-kinase-gamma; MMP-9, metallopeptidase-9; tMCAO, transient occlusion of the middle cerebral artery; WT, wild-type.

Article Snippet: Enzyme-Linked Immunosorbent Assays Enzyme-linked immunosorbent assays of brain cortice homogenates were performed to detect total MMP-9 protein using a Quantikine mouse MMP-9 (total) immunoassay kit (R&D Systems), myeloperoxidase using the mouse Enzyme-linked immunosorbent assay kit (HK210; Hycult Biotechnology), and malondialdehyde (MDA) using the NWLSS MDA assay kit (Northwest Life Science Specialties, LLC) according to the instructions by the manufacturers.

Techniques: Activity Assay, Saline, Control, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Aqueous Artemisia argyi extract mitigates acute lung injury in association with coordinated alterations in gut microbiota, metabolic homeostasis, and pulmonary inflammatory gene expression

doi: 10.3389/fimmu.2026.1770675

Figure Lengend Snippet: AEAA pretreatment is associated with reduced pathological features of LPS-induced acute lung injury in mice. (A) Experimental design of the LPS-induced acute lung injury (ALI) model. (B) Histopathological evaluation of lung tissues. Representative H&E-stained sections from different treatment groups at 20× and 40× magnification show alveolar wall thickening, inflammatory cell infiltration, hemorrhage, and edema in LPS-treated mice. AEAA treatment dose-dependently ameliorated these pathological alterations (red boxed areas). (C) Wet-to-dry (W/D) lung weight ratio. (D) Lung injury score. (E–G) Pro-inflammatory cytokines in serum and BALF.IL-1β (E) , IL-6 (F) , and IL-10 (G) levels in serum and BALF. (H) TNF-α levels in serum and BALF. (I) Myeloperoxidase (MPO) activity. Data are presented as mean ± SEM. n = 5 per group. Statistical analysis was performed using one-way ANOVA followed by post hoc tests. * P < 0.05 , ** P < 0.01 , *** P < 0.001 , **** P < 0.0001 .

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) kits for mouse IL-1β (Cat. #EK201B), IL-6 (Cat. #EK206), IL-10 (Cat. #EK210), TNF-α (Cat. #EK282), and myeloperoxidase (MPO) (Cat. #EK2133) were obtained from Multisciences (Lianke) Biotech Co., Ltd. (Hangzhou, China).

Techniques: Staining, Activity Assay

Journal: Biomedicines

Article Title: SARS-CoV-2 Dysregulates Neutrophil Degranulation and Reduces Lymphocyte Counts

doi: 10.3390/biomedicines10020382

Figure Lengend Snippet: SARS-CoV-2-infected Calu-3 cells secrete factors that diminish neutrophil MPO release. ( A ) Viral titer in the cells and supernates of SARS-CoV-2-infected Calu-3 48 h post infection; UV-irradiated Calu-3 supernates, referred to as Calu-3-conditioned media, were used to treat human neutrophils. ( B ) MPO levels in the treated neutrophils supernatant media determined through ELISA; ( C ) Percentage of neutrophil elastase-positive cells among total cells; ( D ) Representative images at 40× magnification of neutrophils stained for their nucleus (blue), cytoplasm (red), and neutrophil elastase (green); ( E ) Neutrophil elastase levels in the treated neutrophils supernatant media determined through ELISA. Data shown are mean ± SEM; n = 3 per group in each experiment per donor; experiments were repeated twice with two different donors; * p < 0.05, ** p < 0.01, *** p < 0.001 (one-way ANOVA with Tukey’s post hoc test).

Article Snippet: Human MPO ELISA Kit (Abcam, Cambridge, UK) and Human Neutrophil Elastase ELISA Kit (Abcam, Cambridge, UK) were used according to manufacturer’s instructions.

Techniques: Infection, Irradiation, Enzyme-linked Immunosorbent Assay, Staining

Journal: Biomedicines

Article Title: SARS-CoV-2 Dysregulates Neutrophil Degranulation and Reduces Lymphocyte Counts

doi: 10.3390/biomedicines10020382

Figure Lengend Snippet: Direct infection of neutrophils with SARS-CoV-2 promotes CD16 shedding without increasing MPO or elastase release. ( A ) Viral titer in the neutrophils 2 h post infection; ( B ) Percentage of CD16 cells among CD11b+ CD66b+ CD14− CD15+ population (left). Representative flow cytometry density plots of CD11b+ CD66b+ CD14− CD15+ CD16− cells in different treatment groups (right); Levels of MPO ( C ) and neutrophil elastase ( D ) in the supernatant media determined through ELISA. Data shown are mean ± SEM; n = 3 per group in each experiment per donor; experiments were repeated three times with three different donors; * p < 0.05, ** p < 0.01, *** p < 0.001 (Student’s t-test or one-way ANOVA with Tukey’s post hoc test).

Article Snippet: Human MPO ELISA Kit (Abcam, Cambridge, UK) and Human Neutrophil Elastase ELISA Kit (Abcam, Cambridge, UK) were used according to manufacturer’s instructions.

Techniques: Infection, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Didymin Suppresses Microglia Pyroptosis and Neuroinflammation Through the Asc/Caspase-1/GSDMD Pathway Following Experimental Intracerebral Hemorrhage

doi: 10.3389/fimmu.2022.810582

Figure Lengend Snippet: Didymin ameliorated microglial pyroptotic molecules and inflammatory cytokines after ICH. (A) Representative Western blot bands and densitometric quantification of Rkip, Asc, Nlrp3, Caspase-1, and GSDMD. n = 6 for each group. * p < 0.05 vs. sham; # p < 0.05 vs. ICH+vehicle. (B) IL-1β, TNF-α, and MPO production were detected by ELISA assay kit. n = 6 for each group. * p < 0.05 vs. sham; # p < 0.05 vs. ICH+vehicle. (C) Caspase-1/TMEM119 double immunofluorescence staining and quantitative analysis of Caspase-1-positive microglia. Scale bar = 50 μm. n = 3 for each group. * p < 0.05 vs. sham; # p < 0.05 vs. ICH+vehicle.

Article Snippet: A Mouse DuoSet ELISA Kit to IL-1β, TNF-α, and MPO was purchased from R&D Systems and performed as instructed by the manufacturer.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Double Immunofluorescence Staining

Journal: Frontiers in Immunology

Article Title: Didymin Suppresses Microglia Pyroptosis and Neuroinflammation Through the Asc/Caspase-1/GSDMD Pathway Following Experimental Intracerebral Hemorrhage

doi: 10.3389/fimmu.2022.810582

Figure Lengend Snippet: Locostatin abolished the neuroprotective effects of Didymin. (A–C) Neurological deficits, brain water content, and EB extravasation were tested at 24 h after ICH. (D, E) Representative Western blot bands and densitometric quantification of Asc, Nlrp3, Caspase-1, and GSDMD. n = 6 for each group. * p < 0.05 vs. sham; # p < 0.05 vs. ICH+vehicle. & p < 0.05 vs. ICH+Didymin. (F) IL-1β, TNF-α, and MPO production was detected by ELISA assay kit. n = 6 for each group. * p < 0.05 vs.sham; # p < 0.05 vs. ICH+vehicle. & p < 0.05 vs. ICH+Didymin.

Article Snippet: A Mouse DuoSet ELISA Kit to IL-1β, TNF-α, and MPO was purchased from R&D Systems and performed as instructed by the manufacturer.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Didymin Suppresses Microglia Pyroptosis and Neuroinflammation Through the Asc/Caspase-1/GSDMD Pathway Following Experimental Intracerebral Hemorrhage

doi: 10.3389/fimmu.2022.810582

Figure Lengend Snippet: VX-765 ameliorated microglial pyroptosis and brain injury after Locostatin-treated ICH. (A–D) Neurological deficits, brain water content, EB extravasation, and hematoma size were tested at 24 h after ICH. (E) Representative Western blot bands and densitometric quantification of Asc, Nlrp3, Caspase-1, and GSDMD. n = 6 for each group. * p < 0.05 vs. sham; # p < 0.05 vs. ICH+vehicle. (F) IL-1β, TNF-α, and MPO production was tested by ELISA assay. n = 6 for each group. * p < 0.05 vs. sham; # p < 0.05 vs. ICH+vehicle. (G) GSDMD/TMEM119 double immunofluorescence staining and quantitative analysis of GSDMD-positive microglia. Scale bar = 50 μm. n = 3 for each group. * p < 0.05 vs. sham; # p < 0.05 vs. ICH+vehicle.

Article Snippet: A Mouse DuoSet ELISA Kit to IL-1β, TNF-α, and MPO was purchased from R&D Systems and performed as instructed by the manufacturer.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Double Immunofluorescence Staining

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Pterostilbene 4′- β -Glucoside Attenuates LPS-Induced Acute Lung Injury via Induction of Heme Oxygenase-1

doi: 10.1155/2018/2747018

Figure Lengend Snippet: 4-PG and PTER exhibit therapeutic effects in LPS- or P. aeruginosa -induced acute lung injury. (a–g) C57BL/6 mice were challenged with LPS (2.5 mg/kg) by intranasal exposure for 24 h. 4-PG (10 mg/kg per day, i.p.) and PTER (10 mg/kg per day, i.p.) were administered 5 h after LPS challenge. (b) Representative lung histology was shown by hematoxylin and eosin- (H&E-) stained lung sections from six experimental groups (left). Quantitative analysis of histologic lung section by lung injury score for six experimental groups. The score generates the average of two independent investigators (right). (c and d) BAL fluid and lung tissues were harvested after treatment with LPS for 24 h and then (c) analyzed for total cells in BAL fluid. (d) Neutrophil infiltration into lung tissues was measured by activity of myeloperoxidase (MPO) in lung tissues. (e and f) The secreted levels of TNF- α and IL-6 in BAL fluid were analyzed by ELISA, respectively. (g) The mRNA levels of TNF- α , IL-6, IL-1 β , CXCL1, and CXCL2 in lung tissues were detected by RT-PCR. (h–m) C57BL/6 mice were instilled P. aeruginosa (1 × 10 7 CFU/mouse) by intranasal exposure for 24 h. After instillation of P. aeruginosa , mice were post-treated with 4-PG (10 mg/kg, i.p.) at 6, 12, and 18 h, respectively. After 24 h, BAL fluid and lung tissues were collected and then (i) analyzed for total cells in BAL fluid. (j) Neutrophil infiltration into lung tissues was analyzed by MPO in lung tissues. (k and l) The levels of TNF- α and IL-6 in BAL fluid were analyzed by ELISA, respectively. (m) The wet-to-dry weight ratio of whole lungs was determined on 24 h after the stimulation of P. aeruginosa . Data were expressed as means ± SD, n = 3. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: To measure neutrophil infiltration into lung tissues, MPO enzyme activity in lung tissues was measured using the Mouse Myeloperoxidase DuoSet kit (R&D Systems, Minneapolis, MN).

Techniques: Staining, Activity Assay, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of Clinical and Translational Hepatology

Article Title: Single-cell Sequencing Reveals Neutrophil Extracellular Traps in Association with Endotheliopathy and Immunothrombosis in Hepatitis B Virus-related Acute-on-chronic Liver Failure

doi: 10.14218/JCTH.2025.00666

Figure Lengend Snippet: (A) Plasma concentrations of NETs markers, including MPO–DNA complexes, MPO, and NE, in HBV-ACLF, CHB, and HC groups. (B–C) Correlations between plasma MPO–DNA levels and INR and TAT (B), and LDH (C). (D) Correlations between plasma MPO–DNA levels and established prognostic scoring systems, including the COSSH-ACLF II and MELD-Na scores. (E) Comparison of NETs marker levels (MPO–DNA and MPO) between survivors and non-survivors within the HBV-ACLF cohort. (F) Kaplan–Meier survival curves of HBV-ACLF patients stratified by plasma MPO–DNA levels (≤2-fold vs >2-fold). *** P < 0.001, and **** P < 0.0001. NETs, neutrophil extracellular traps; HBV-ACLF, hepatitis B virus-related acute-on-chronic liver failure; MPO, myeloperoxidase; NE, neutrophil elastase; CHB, chronic hepatitis B; HC, healthy controls; INR, international normalized ratio; TAT, thrombin-antithrombin complex; LDH, lactate dehydrogenase; COSSH-ACLF II, Chinese Onset Study of Severe Hepatitis with Acute-on-Chronic Liver Failure II; MELD-Na, Model for End-Stage Liver Disease-Sodium.

Article Snippet: Specifically, the Human Myeloperoxidase ELISA Kit (RayBiotech, Cat. No. ELH-MPO) and the Human Neutrophil Elastase ELISA Kit (RayBiotech, Cat. No. ELH-NEUTRO-1) were used according to the manufacturers’ protocols.

Techniques: Clinical Proteomics, Comparison, Marker, Virus

Journal: Journal of Clinical and Translational Hepatology

Article Title: Single-cell Sequencing Reveals Neutrophil Extracellular Traps in Association with Endotheliopathy and Immunothrombosis in Hepatitis B Virus-related Acute-on-chronic Liver Failure

doi: 10.14218/JCTH.2025.00666

Figure Lengend Snippet: (A–C) Correlation analyses between plasma MPO–DNA complex levels and syndecan-1 (A), ICAM-1 (B), and sTM and vWF (C). (D) Immunofluorescence staining of endothelial cells (CD31), NETs (citrullinated histone H3, Cit-H3), and platelets (CD41) in liver tissues from patients with HBV-ACLF, CHB, and HC. Scale bars: 50 µm. (E) Neutrophils isolated from CHB patients were stimulated with PMA. NETs were collected and incubated with human aortic endothelial cells, with or without DNase I treatment. Syndecan-1 levels in supernatants and plasma clotting time of cells were measured after 6 h. ** P < 0.01, *** P < 0.001, and **** P < 0.0001. NETs, neutrophil extracellular traps; HBV-ACLF, hepatitis B virus-related acute-on-chronic liver failure; MPO, myeloperoxidase; ICAM-1, intercellular adhesion molecule-1; sTM, soluble thrombomodulin; vWF, von Willebrand factor; CHB, chronic hepatitis B; HC, healthy controls; PMA, phorbol 12-myristate 13-acetate; DNase I, deoxyribonuclease I.

Article Snippet: Specifically, the Human Myeloperoxidase ELISA Kit (RayBiotech, Cat. No. ELH-MPO) and the Human Neutrophil Elastase ELISA Kit (RayBiotech, Cat. No. ELH-NEUTRO-1) were used according to the manufacturers’ protocols.

Techniques: Clinical Proteomics, Immunofluorescence, Staining, Isolation, Incubation, Coagulation, Virus

Journal: Mediators of Inflammation

Article Title: Lipopolysaccharide-Binding Protein Downregulates Fractalkine through Activation of p38 MAPK and NF- κ B

doi: 10.1155/2017/9734837

Figure Lengend Snippet: LBP gene overexpression further decreased LPS-induced downregulation of FKN mRNA and protein expression; LBP gene silencing, SB203580, and SC-514 suppressed LPS-induced downregulation of FKN mRNA and protein expression, respectively, in A549 cells. RT-PCR and ELISA were used to analyze the LBP and FKN mRNA and protein expression, respectively, in A549 cells. The LBP mRNA expression is shown in (a). The relative LBP mRNA levels (normalized to GAPDH mRNA) in LPS-stimulated cells are significantly higher than those in control cells ( p < 0.05). The LBP mRNA levels of the LBP(+) group and LBP(−) group are upregulated and downregulated, respectively, when compared with the control group ( p < 0.05 in all cases). The LBP mRNA levels of the LPS+LBP(+) group of cells are upregulated compared with those of the control group and LPS group ( p < 0.05 in all cases). The LBP mRNA levels of the LPS+LBP(−) group of cells are upregulated compared with those of the control group and downregulated compared with those of the LPS group ( p < 0.05 in all cases). The LBP mRNA level of the empty vector group is not different from that of the control group ( p > 0.05). The FKN mRNA expression is shown in (e). The relative FKN mRNA levels (normalized to GAPDH mRNA) in LPS-stimulated cells are significantly lower than those in control cells ( p < 0.05). The FKN mRNA levels of the LPS+LBP(+) group of cells are downregulated compared with those of the control group and the LPS group ( p < 0.05 in all cases). The FKN mRNA levels of the LPS+LBP(−), LPS+SB, and LPS+SC groups of cells are downregulated compared with those of the control group and upregulated compared with those of the LPS group ( p < 0.05 in all cases). The FKN mRNA levels of the TNF- α group are upregulated compared with those of the control group ( p < 0.05). The results of ELISA and western blotting in (b), (c), and (d) demonstrate that the LPS treatment significantly increases the levels of LBP protein expression ( p < 0.05). The LBP protein levels of the LBP(+) group and LBP(−) group are upregulated and downregulated, respectively, compared with those of the control group ( p < 0.05 in all cases). The LBP protein levels of the LPS+LBP group of cells are upregulated compared with those of the control group and the LPS group ( p < 0.05 in all cases). The LBP protein levels of the LPS+LBP(−) group of cells are upregulated compared with those of the control group and downregulated compared with those of the LPS group ( p < 0.05 in all cases). ELISA and western blotting assay in (f), (g), and (h) demonstrate that the LPS treatment significantly reduces the levels of FKN protein expression ( p < 0.05). The FKN protein levels of the LPS+LBP group of cells are downregulated compared with those of the control group and the LPS group ( p < 0.05 in all cases). The FKN protein levels of the LPS+LBP(−), LPS+SB, and LPS+SC groups of cells are downregulated compared with those of the control group and upregulated compared with those of the LPS group ( p < 0.05 in all cases). The FKN protein levels of the TNF- α group are higher than those of the control group ( p < 0.05). Immunofluorescence staining results in (i) and (j) confirmed that TNF- α induced a high FKN-positive cell ratio in A549 cells. LPS decreased FKN-positive cell ratio in A549 cells; LBP gene overexpression further decreased FKN-positive cell ratio which was inhibited by LPS, LBP gene silencing, and SB203580. Also, SC-514 reduced the inhibition of LPS ( p < 0.05 in all cases). The photomicrographs are 200x magnification. Data are presented as means ± SD of six independent experiments ( n = 6). ∗ represents p < 0.05 when compared with that in the CTL; # represents p < 0.05 when compared with that in the LPS; △ represents p < 0.05 when compared with that in the LPS+LBP(+). ∗ p < 0.05 when compared with that in the control group; ◇ represents p < 0.05 when compared with that in the LPS+LBP(−) group; △ represents p < 0.05 when compared with that in the LPS+LBP(+).

Article Snippet: The levels of myeloperoxidase activity were measured by the rat MPO ELISA kit (HK105-01, Hycult Biotech, USA) according to the manufacturer's instructions.

Techniques: Over Expression, Expressing, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control, Plasmid Preparation, Western Blot, Immunofluorescence, Staining, Inhibition

Journal: Mediators of Inflammation

Article Title: Lipopolysaccharide-Binding Protein Downregulates Fractalkine through Activation of p38 MAPK and NF- κ B

doi: 10.1155/2017/9734837

Figure Lengend Snippet: SB203580 and SC-514 ameliorated LPS-induced lung injury and inflammation. Hematoxylin and eosin staining was used to determine the LPS-induced lung injury and inflammatory response in the lung tissue of the ARDS rat model. (a) The control group rats (A) had a distinct framework with complete alveolar walls and interstitium without exfiltration. The LPS group rats (B) showed edema, neutrophil infiltration, hemorrhage, moderate bronchiole epithelial desquamation, and minimal hyaline membrane formation. Histotological analysis also show that rats receiving LBPK95A, SB203508, or SC514 injection had substantially less inflammatory cell infiltration, edema, hemorrhage, and thickness of alveolar walls in comparison to rats treated with LPS ((C), (D), and (E), resp.). The photomicrographs are at 400x magnification. ((b) and (c)) The total number of cells, neutrophil number, and neutrophil ratio in BALF were higher in the LPS group than that in the control group and lower in the LPS+LBPK95A, LPS+SB, and LPS+SC groups than that in the LPS group ( p < 0.05, n = 10, in all cases). The MPO of lung tissues and BALF was determined by ELISA. The results show that the level of MPO concentration in the LPS group was significantly higher than that in the control group ( p < 0.05). (d) However, the MPO concentration level in the LPS+LBPK95A, LPS+SB, and LPS+SC groups was significantly lower than that in the LPS group ( p < 0.05 in all cases). (e) The LPS group had a significantly higher W/D ratio than the control group, and the W/D ratio in the LPS+LBPK95A, LPS+SB, and LPS+SC groups were significantly lower than that in the LPS group ( p < 0.05 in all cases). The LPS group had a lower PaO 2 /FiO 2 ratio than the control group. (f) The LPS+LBPK95A, LPS+SB, and LPS+SC groups had higher PaO 2 /FiO 2 ratios than the LPS group ( p < 0.05 in all cases). ∗ represents p < 0.05 when compared with that in the CTL group of rats; # represents p < 0.05 when compared with that in the LPS group of rats; △ represents p < 0.05 when compared with that in the LPS+LBPK95A group of rats ( n = 10).

Article Snippet: The levels of myeloperoxidase activity were measured by the rat MPO ELISA kit (HK105-01, Hycult Biotech, USA) according to the manufacturer's instructions.

Techniques: Staining, Control, Membrane, Injection, Comparison, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Mediators of Inflammation

Article Title: Lipopolysaccharide-Binding Protein Downregulates Fractalkine through Activation of p38 MAPK and NF- κ B

doi: 10.1155/2017/9734837

Figure Lengend Snippet: Expression of LBP and FKN protein in lung tissue homogenates and in serum of the ARDS rat model. The LBP and FKN expression in lung tissue homogenates and sera of rats with ARDS was determined by ELISA and western blotting. The results demonstrate that (a) LPS increased the levels of LBP expression significantly in both cases ( p < 0.05 in all cases). The LBP levels of the LPS+LBPK95A group were upregulated compared with those of the control group and downregulated compared with those of the LPS group ( p < 0.05 in all cases) ((a), (b), and (c)). (b) LPS reduced the levels of FKN expression in both cases ( p < 0.05 in all cases). The FKN levels of the LPS+SB and LPS+SC groups are downregulated compared with those of the control group and upregulated compared with those of the LPS group ( p < 0.05 in all cases) ((d), (e), and (f)). The FKN levels of the LPS+SB and LPS+SC groups are lower than those of the LPS+LBPK95A group ( p < 0.05 in all cases) ((d), (e), and (f)). ∗ represents p < 0.05 when compared with that in the CTL group rats; # represents p < 0.05 when compared with that in the LPS group rats. △ represents p < 0.05 when compared with that in the LPS+LBPK95A rats. Immunohistochemical staining was used to determine the localization of FKN in the rat lungs. As can be seen, the major sites of FKN expression are in alveolar type I and type II epithelia. The expression of FKN protein was compared between the different groups of rats. The staining intensity and number of cells expression of FKN decreased markedly in the LPS group, when compared to those of the control group. The use of LBPK95A, SB203580, and SC-514 partly reduced the reduction of FKN seen (g). The photomicrographs are at 400x magnification.

Article Snippet: The levels of myeloperoxidase activity were measured by the rat MPO ELISA kit (HK105-01, Hycult Biotech, USA) according to the manufacturer's instructions.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Control, Immunohistochemical staining, Staining

Journal: Clinical and Translational Medicine

Article Title: Novel bispecific nanobody mitigates experimental intestinal inflammation in mice by targeting TNF‐α and IL‐23p19 bioactivities

doi: 10.1002/ctm2.1636

Figure Lengend Snippet: Amelioration of inflammatory symptoms in DSS‐induced colitis mice by antibody treatment. (A) Experimental protocol illustrating the induction of DSS‐induced colitis in mice. Mice ( n = 5) were administered DSS (3%) in their drinking water for 7 days, followed by treatment with antibodies (BsNb, BsNb‐Fc, IFX, VHH#2 and VHH#37) at a dose of 5 mg kg −1 body weight for 7 days. (B) Body weight variation curves depicting the changes in body weight of mice treated with antibodies compared to the controls. (C) Disease activity index (DAI) scores of the different treatment groups over time, reflecting the severity of colitis. (D) Myeloperoxidase (MPO) activity in different treatment group, with MPO levels in the control group considered as the reference unit. (E and F) Evaluation of colon length and spleen weight in mice subjected to DSS and antibody treatment on day 15. Representative images and measurements of colon and spleen length and morphology were shown. (G) Representative histological images of colon tissues stained with H&E, with corresponding calculation of pathological score to assess colonic tissue damage. Scale bars, 50 µm. (H, I and J) Immunohistochemical (IHC) staining and quantitation of CD4 + , F4/80 + and MPO + in the proximal colon of controls, DSS‐treated, BsNb‐Fc and IFX‐treated mice. Scale bars, 50 µm. (K) Measurement of cytokine expression levels in sera of controls, DSS‐treated, BsNb‐Fc and IFX‐treated mice using ELISA. All data represented as mean ± SEMs; * p < .05; ** p < .01; *** p < .001; **** p < .0001.

Article Snippet: MPO activity was measured using an MPO activity assay kit (E‐BC‐F013, Elabscience).

Techniques: Activity Assay, Control, Staining, Immunohistochemical staining, Immunohistochemistry, Quantitation Assay, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Clinical and Translational Medicine

Article Title: Novel bispecific nanobody mitigates experimental intestinal inflammation in mice by targeting TNF‐α and IL‐23p19 bioactivities

doi: 10.1002/ctm2.1636

Figure Lengend Snippet: BsNb‐Fc demonstrates superior efficacy compared to IFX, UST and IFX&UST combination in a TNBS‐induced mouse colitis model. (A) Experimental protocol for TNBS‐induced colitis in mice. Mice ( n = 5) were sensitized with TNBS (1%) on the abdominal skin for 7 days, followed by intrarectal administration of 2.5% TNBS. Subsequently, the mice were treated with the following antibodies: BsNb‐Fc (5 mg/kg), IFX (2.5 mg/kg) &UST (.25 mg/kg), IFX (5 mg/kg), UST (.5 mg/kg) and isotype antibody for 7 days. (B) Body weight variation of antibodies‐treated mice compared to controls. (C) MPO activity of different treatment groups using an MPO activity assay kit. (D) Assessment of colonic morphology and length in mice treated with TNBS and antibodies. (E) Representative histological images of colon tissues stained with H&E, and calculation of the pathological score of colonic tissues. Scale bars, 50 µm. (F) Measurement of cytokine expression levels in colon tissues using ELISA. All data represented as mean ± SEMs. Statistical significance: * p < .05; ** p < .01; *** p < .001; **** p < .0001.

Article Snippet: MPO activity was measured using an MPO activity assay kit (E‐BC‐F013, Elabscience).

Techniques: Activity Assay, Staining, Expressing, Enzyme-linked Immunosorbent Assay