Journal: Thrombosis Journal

Article Title: Complement C5a induces the generation of neutrophil extracellular traps by inhibiting mitochondrial STAT3 to promote the development of arterial thrombosis

doi: 10.1186/s12959-022-00384-0

Figure Lengend Snippet: C5a promoted arterial thrombosis by triggering neutrophils to release NETs. A , B H&E staining of thrombus, cross-sections ( A ) and longitudinal sections ( B ). Scale bar = 100 µm. C Quantification of the thrombus size in cross-sections by H&E staining ( n = 5 each). D Thrombus weight ( n = 5 each). E The blood flow velocity in the LICA was dramatically decreased in the FeCl 3 -induced thrombus mice compared with the sham mice, and this change was reversed by PMX53. F Quantification of the blood flow velocity ( n = 4 each). G Immunofluorescence staining of cross-sections of the LCCA for CitH3 (red), Ly6g (green) and DAPI (blue) 6 h after FeCl 3 exposure. Thirty minutes before FeCl 3 exposure, PMX53 was administered by intraperitoneal injection. Scale bar = 75 µm. H Quantification of NET formation capacity shown as the ratio of NETs/neutrophil in LCCA cross-sections subjected to immunofluorescence staining (NaCl group = 3, PMX53 group = 4). I Representative images of immunofluorescence stainings of NET formation for DNA (SYTOX green), CitH3 (red), Ly6g(blue) in vitro after stimulation of C5a, and NET formation was attenuated by PMX53. Scale bar = 75 µm. J Quantification of NET formation capacity in vitro shown as the percentage of NET release, which was assessed by immunofluorescence staining ( n = 3 each). (K) NETosis was measured using a plate reader assay. NET release is expressed as percentage of total DNA ( n = 3 each). Data are presented as mean ± SD

Article Snippet: Mouse neutrophils were isolated from the bone marrow of tibias and femurs from healthy C57BL/6 J mice using the Neutrophil Isolation Kit (Solarbio, China) following the manufacturer’s instructions.

Techniques: Staining, Immunofluorescence, Injection, In Vitro

Journal: Thrombosis Journal

Article Title: Complement C5a induces the generation of neutrophil extracellular traps by inhibiting mitochondrial STAT3 to promote the development of arterial thrombosis

doi: 10.1186/s12959-022-00384-0

Figure Lengend Snippet: Interactions among C5a, mitochondrial STAT3 and NETs. A , B Western blot analysis was performed to test the expression levels of mitochondrial STAT3 and p-STAT3 (Ser 727 ) in neutrophil-like cells cocultured with C5a, compared with the control. VDAC, a marker of mitochondria, was used as a loading control for mitochondria ( n = 5 each). C NET release in response to buffer or AG490 was measured using a plate reader assay ( n = 3 each). D Representative images of immunofluorescence staining for DNA (SYTOX green), CitH3 (red) and Ly6g (blue) in vitro after stimulation with AG490 showing the presence of NETs. Scale bar = 100 µm. E Quantification of NET formation capacity shown as the percentage of NET release in vitro after stimulation with buffer or AG490, as assessed by immunofluorescence staining. ( n = 3 each). Data are presented as mean ± SD

Article Snippet: Mouse neutrophils were isolated from the bone marrow of tibias and femurs from healthy C57BL/6 J mice using the Neutrophil Isolation Kit (Solarbio, China) following the manufacturer’s instructions.

Techniques: Western Blot, Expressing, Control, Marker, Immunofluorescence, Staining, In Vitro

Journal: Thrombosis Journal

Article Title: Complement C5a induces the generation of neutrophil extracellular traps by inhibiting mitochondrial STAT3 to promote the development of arterial thrombosis

doi: 10.1186/s12959-022-00384-0

Figure Lengend Snippet: The reduction in arterial thrombotic burden induced by PMX53 was abolished by AG490 in vivo. A , B H&E staining of thrombus, cross-sections A and longitudinal sections B . The thrombus area was reduced by PMX53, and AG490 abolished this effect. A Scale bar = 100 µm; B Scale bar = 200 µm. C Quantification of the thrombus size in cross-sections by H&E staining ( n = 5 each). D Thrombus weight ( n = 5 each). E Blood flow velocity in the LICA. AG490 reversed the increased in blood flow induced by PMX53. F Quantification of the blood flow velocity ( n = 4 each). G Immunofluorescence staining of cross-sections of the LCCA for CitH3 (red), Ly6g (green) and DAPI (blue) 6 h after FeCl 3 exposure. Scale bar = 75 µm. H Quantification of NET formation capacity shown as the ratio of NETs/neutrophil in LCCA cross-sections subjected to immunofluorescence staining (NaCl group = 3; PMX53 group = 4; PMX53 + AG490 group = 3; AG490 group = 4). Data are presented as mean ± SD

Article Snippet: Mouse neutrophils were isolated from the bone marrow of tibias and femurs from healthy C57BL/6 J mice using the Neutrophil Isolation Kit (Solarbio, China) following the manufacturer’s instructions.

Techniques: In Vivo, Staining, Immunofluorescence

Journal: Thrombosis Journal

Article Title: Complement C5a induces the generation of neutrophil extracellular traps by inhibiting mitochondrial STAT3 to promote the development of arterial thrombosis

doi: 10.1186/s12959-022-00384-0

Figure Lengend Snippet: Visual summary: the effect of complement C5a in arterial thrombosis. In arterial thrombosis, C5a chemotactically attracts neutrophils to migrate towards the culprit site and triggers the release of NETs, which contribute to thrombosis by promoting coagulation and stabilizing clots. C5a-induced promotion of NET release is dependent on Mito-ROS production. C5a induces the Mito-ROS production by inhibiting mitochondrial STAT3 activity

Article Snippet: Mouse neutrophils were isolated from the bone marrow of tibias and femurs from healthy C57BL/6 J mice using the Neutrophil Isolation Kit (Solarbio, China) following the manufacturer’s instructions.

Techniques: Coagulation, Activity Assay

Journal: British journal of pharmacology

Article Title: Salvianolic acid B improves the microcirculation in a mouse model of sepsis through a mechanism involving the platelet receptor CD226.

doi: 10.1111/bph.17371

Figure Lengend Snippet: FIGURE 3 SalB inhibits platelet adhesion. (a) Mouse platelets were collected 12 h after modelling, and the adhesion of MPVECs to platelets was detected by Dil fluorescent probe staining. N = 5; scale bars = 150 μm. (b) Fluorescence intensity value statistics. N = 5. (c) Neutrophil– platelet, monocyte–platelet and lymphocyte–platelet aggregations were detected in mouse peripheral blood 12 h after modelling. N = 6. (d) The peripheral neutrophils were collected 12 h after modelling, and the adhesion between platelets and neutrophils was observed by scanning electron microscopy. Sham + saline: the sham group was given saline; CLP + saline: the sepsis group was injected i.p.with saline; CLP + SalB: the sepsis group was injected i.p. with SalB. Data shown are individual values with means ± SEM. #P < 0.05, significantly different from sham + saline group. *P < 0.05, significantly different from CLP + saline groups.

Article Snippet: Neutrophils from mice were initially enriched using a peripheral blood neutrophil isolation kit (Solarbio, Beijing, China) and sorted using a FACSAria IIu cytometer (BD, San Jose, CA, USA), resulting in a purity of ≥98%.

Techniques: Staining, Fluorescence, Electron Microscopy, Saline, Injection

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: miR-30c-5p Reduces Renal Ischemia-Reperfusion Involving Macrophage

doi: 10.12659/MSM.914579

Figure Lengend Snippet: MiR-30c-5p attenuated rat renal I/R and inhibited serum Cr and serum NGAL. After the rats had surgery to create a rate renal I/R model, the rats were fed normally. Serum samples were taken at 24 hours, 48 hours, and 96 hours after normal feeding. Serum Cr was measured by serum Cr assay kit using automatic chemical analysis at the wavelength of 570 nm ( A ). Serum NGAL was evaluated by ELISA kit ( B ). After the rats were sacrificed, the kidney tissues were removed, and the kidney injury was measured by H&E staining ( C ). MiR-30=miR-30c-5p. The values were presented as mean ±SD, and the data were analyzed by ANOVA with Turkey’s multiple test (a versus Sham group, b versus I/R group; a=b P <0.05, aa=bb P <0.01). I/R – ischemia-reperfusion; Cr – creatinine; NGAL – neutrophil gelatinase associated lipocalin; ELISA – enzyme-linked immunosorbent assay; H&E – hematoxylin and eosin; SD – standard deviation; ANOVA – analysis of variance.

Article Snippet: Rat neutrophil gelatinase associated lipocalin (NGAL) ELISA kit (Elabscience, Wuhan, Hubei, China) measured the concentration of NGAL in blood samples with MultiskanTM FC (Thermo Scientific, Waltham, MA, USA) at the wavelength of 450 nm, following the product’s specifications.

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Standard Deviation

Journal: Respiratory research

Article Title: Neutrophil extracellular traps promote proliferation of pulmonary smooth muscle cells mediated by CCDC25 in pulmonary arterial hypertension.

doi: 10.1186/s12931-024-02813-2

Figure Lengend Snippet: Fig. 1 Bioinformatics analysis of patients with or without PAH. (A) Microarray data difference analysis volcano map. (B) Clustering heatmap of the top 50 differential genes in the microarray data. (C) Single-cell sequencing data, subpopulation grouping UMAP. (D) Single-cell sequencing data, cell proportion map of each sample. (E) Single-cell sequencing data, neutrophil grouping UMAP. (F) Single-cell sequencing data, PASMCs grouping UMAP

Article Snippet: Neutrophils were isolated using a Rat Neutrophil Isolation Kit (Solarbio, China).

Techniques: Microarray, Sequencing

Journal: Cell Reports Medicine

Article Title: A functional cardiac patch promotes cardiac repair by modulating the CCR2 − cardiac-resident macrophage niche and their cell crosstalk

doi: 10.1016/j.xcrm.2025.101932

Figure Lengend Snippet:

Article Snippet: Rat CXCL2/MIP-2 ELISA Kit , Boster Bio. , EK0725.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Staining, Labeling, H&E Stain, Lysis, Software