Journal: Nanoscale

Article Title: Differentiating sepsis from non-infectious systemic inflammation based on microvesicle-bacteria aggregation.

doi: 10.1039/c5nr01851j

Figure Lengend Snippet: Figure 1: Trigger-dependent Microvesicle Shedding. Scanning electron micrograph (a) and size-distribution assessed by NTA (b) of PMN-derived microvesicles originating from PMNs incubated with plasma-opsonized S. aureus bacteria, E. coli, LPS, heat-inactivated bacteria bioparticles or vehicle (HBSS). PMN-derived CD11β/CD18 and CD11β/CD177-double positive events assessed by flow cytometry as a function of bacterial triggering agent (n = 3) (c). Scanning (d,e) and transmission electron micrographs (f,g) of PMNs showing pronounced membrane budding and shedding of microvesicles following incubation with opsonised S. aureus particles for 30 minutes (arrow indicates S. aureus particle) (e,g) compared to PMNs incubated with HBSS (d,f). 3D-tomographies and outer surface reconstructions of PMN incubated with S. aureus further confirmed the constriction of vesicles from the outer membrane seen in TEM (h). Raman spectroscopy maps of PMN incubated with (top, I) or without (bottom, II, control) bacteria showed lipid droplets and peri-membranous accumulation of glycogen granules in stimulated PMNs (I) compared to control (II) (i). PMNs exposed to S. aureus compared to resting PMNs (Figure 1d,e). Transmission electron micrographs of thin sections of PMNs containing phagocytised S. aureus bacteria confirmed increased membrane budding and formation of microvesicles (Figure 1f,g). Formation of glycogen granule clusters, translocation and peri- membranous massing of glycogen granule aggregates, and shipping of cytoplasmatic microvesicles containing glycogen granules were observed in PMNs exposed to bacteria, while glycogen granules remained well-dispersed in the cytoplasm of unstimulated PMNs (Figure 1f,g). 3D-tomography of PMNs further confirmed

Article Snippet: For flow cytometry, FITC mouse anti-rat CD18 (WT3, IgG1, AbD Serotec) and Alexa-647 anti-rat CD11β (OX-42, IgG2a, κ, BioLegend) were used for double staining at a concentration of 1 μg mL−1.

Techniques: Derivative Assay, Incubation, Clinical Proteomics, Bacteria, Flow Cytometry, Transmission Assay, Membrane, Raman Spectroscopy, Control, Translocation Assay, Tomography

Journal: Nanoscale

Article Title: Differentiating sepsis from non-infectious systemic inflammation based on microvesicle-bacteria aggregation.

doi: 10.1039/c5nr01851j

Figure Lengend Snippet: Figure 3. Microvesicles in Plasma Samples from an Experimental Sepsis Model. Caecal ligation and puncture (CLP) procedure in rats (a). Time-dependent concentration of neutrophil- derived CD11β/CD18-double positive microvesicles assessed by flow cytometry (b). Aggregation of S. aureus bacteria standard with microvesicle isolates from animal plasma at the 24 and 48 hour time point (c) and corresponding ROC curves (d). Characterization of Microvesicle-Bacteria Aggregates In order to better understand the nature of the microvesicle-bacteria aggregates, we used an in vitro analysis to further characterize their properties. The CD11β-positivity of the aggregating human PMN- derived vesicles was confirmed by immunostaining (Figure 4a) and transmission electron micrographs of microvesicle-bacteria aggregates were recorded (Figure 4b). The microvesicle- concentration dependence of bacteria aggregation was confirmed by serially diluting microvesicle isolates from PMNs exposed to S.

Article Snippet: For flow cytometry, FITC mouse anti-rat CD18 (WT3, IgG1, AbD Serotec) and Alexa-647 anti-rat CD11β (OX-42, IgG2a, κ, BioLegend) were used for double staining at a concentration of 1 μg mL−1.

Techniques: Clinical Proteomics, Ligation, Concentration Assay, Derivative Assay, Flow Cytometry, Bacteria, In Vitro, Immunostaining, Transmission Assay

Journal:

Article Title: Effect of notoginsenoside R1 on hepatic microcirculation disturbance induced by gut ischemia and reperfusion

doi: 10.3748/wjg.14.29

Figure Lengend Snippet: Effect of R1 on the expression of E-selectin in hepatic vessels (A) and CD18 and CD11b in neutrophils (B) of mice subjected to SMA I/R. The results are presented as mean ± SE from 6 animals. aP < 0.05 vs control, cP < 0.05 vs I/R.

Article Snippet: Other reagents used in experiments were as follows: rhodamine 6G (purity > 99.0%, Lot No.2350994, Fluka Co., Switzerland), FITC-rat anti-mouse CD18 monocolonal antibody (Lot No.553293, BD Biosciences PharMingen, USA), FITC-rat anti-mouse CD11b monoclonal antibody (Lot No.557396, BD Biosciences PharMingen, USA), goat polyclonal antibody against mouse E-selectin (M-20) (sc-6939, Santa Cruz Biotechnology, Inc. USA), goat polyclonal antibody against mouse ICAM-1 (M-19) (sc-1511, Santa Cruz Biotechnology, Inc. USA), rhodamine conjugated rabbit anti-goat lgG-R (Lot No.B1006, Santa Cruz Biotechnology, Inc. USA), Hoechst33342 (Lot No.6538, Santa Cruz Biotechnology, Inc. USA), mouse MCP-1 flex set (Lot No.558342, BD Biosciences, USA), mouse TNF flex set (Lot No.558299, BD Biosciences, USA), mouse IL-6 flex set (Lot No.558301, BD Biosciences, USA).

Techniques: Expressing

Journal: Cell & Bioscience

Article Title: Tie2-expressing monocytes/macrophages promote angiogenesis in chronically ischaemic brain tissue

doi: 10.1186/s13578-025-01401-1

Figure Lengend Snippet: Immunofluorescence and western blotting results showing the expression of relevant cytokines in the CIBT of 2VO + EMS rats. A Representative triple immunofluorescence staining images showing ANGPT2 + , Tie2 + , and CD11b + cells in the CIBT in each group (white arrows indicate TEMs). Bar = 50 µm. B Counts of ANGPT2 + cells in each group (n = 3, one-way ANOVA and Šídák's multiple comparisons test). C Counts of TEMs (Tie2 + and CD11b + ) in each group (n = 9, one-way ANOVA and Šídák's multiple comparisons test). D Representative western blot showing the relative expression of TRPS1, ANGPT2, Tie2, CD11b, VEGFA, IGF1, and CD31 in the CIBT adjacent to the TM tissue in each group (normalized to β-actin expression). E Densitometric analyses of the relative expression of TRPS1, ANGPT2, Tie2, CD11b, VEGFA, IGF1, and CD31 (n = 3, one-way ANOVA and Šídák's multiple comparisons test; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001). The error bars represent the ± SDs. CIBT: chronically ischaemic brain tissue; EMS: encephalomyosynangiosis; TEMs: Tie2-expressing monocytes/macrophages; 2VO: 2-vessel occlusion

Article Snippet: After being washed with PBST, the slices were incubated with 488-conjugated goat anti-rabbit (1:2000, ab150077, Abcam, UK), 594-conjugated goat anti-mouse (1:2000, ab150116, Abcam, UK), and 647-conjugated donkey anti-goat (1:2000, ab150135, Abcam, UK) secondary antibodies at room temperature in a dark room for 2 h. The 647-conjugated CD11b antibody (0.5 μg, CL647-65229, proteintech, China) was used to directly label CD11b.

Techniques: Immunofluorescence, Western Blot, Expressing, Staining

Journal: Cell & Bioscience

Article Title: Tie2-expressing monocytes/macrophages promote angiogenesis in chronically ischaemic brain tissue

doi: 10.1186/s13578-025-01401-1

Figure Lengend Snippet: Immunofluorescence results showing the expression of pro-angiogenesis factors in the CIBT of 2VO + EMS rats. A Representative triple immunofluorescence staining images showing Tie2 + , CD11b + , and VEGFA + cells and in the CIBT in each group (white arrows indicate TEMs). Bar = 50 µm. B Counts of VEGF + cells in each group (n = 3, one-way ANOVA and Šídák's multiple comparisons test). C Representative triple immunofluorescence staining images showing Tie2 + , CD11b + , and IGF1 + cells in the CIBT in each group (white arrows indicate TEMs). Bar = 50 µm. D Counts of IGF1 + cells in each group (n = 3, one-way ANOVA and Šídák's multiple comparisons test). CIBT: chronically ischaemic brain tissue; EMS: encephalomyosynangiosis; TEMs: Tie2-expressing monocytes/macrophages; 2VO: 2-vessel occlusion

Article Snippet: After being washed with PBST, the slices were incubated with 488-conjugated goat anti-rabbit (1:2000, ab150077, Abcam, UK), 594-conjugated goat anti-mouse (1:2000, ab150116, Abcam, UK), and 647-conjugated donkey anti-goat (1:2000, ab150135, Abcam, UK) secondary antibodies at room temperature in a dark room for 2 h. The 647-conjugated CD11b antibody (0.5 μg, CL647-65229, proteintech, China) was used to directly label CD11b.

Techniques: Immunofluorescence, Expressing, Staining