Journal: Journal of Inflammation Research

Article Title: Cell Death Index Predicts Sepsis Outcomes and Highlights Necroptosis as a Therapeutic Target

doi: 10.2147/JIR.S577930

Figure Lengend Snippet: Necrostatin-1 significantly improved sepsis outcomes in CLP models. (a) Serum levels of HMGB1 and CASP8 in clinical patients. (b) Schematic of the animal experimental design. (c) Survival rates of CLP mice with or without Nec-1 (10 mg/kg). (d) The mRNA levels of PDZD8 and ADRB2 were analyzed by RT-PCR in the peripheral blood of mice. (e) At the 22h post-CLP, the levels of inflammatory cytokines (TNF-α and IL-6) in plasma were detected by ELISA tests. (f) Hematoxylin–eosin staining of lung, liver, kidney and spleen sections, with red arrows indicating pathological areas. Scale bar = 50 μm. All data were expressed as mean ± SEM (n=3). *P< 0.05, **P< 0.01, ***P< 0.001.

Article Snippet: According to the manufacturer’s protocols, concentrations of HMGB1 (Elabscience, E-EL-H1554, Wuhan, China), CASP8 (Elabscience, E-EL-H0659, Wuhan, China), IL-6 (Solarbio, SEKM-0034, Beijing China), and TNF-α (ABclonal, RK04595, Wuhan, China) were measured using ELISA kits with standard curves ( Table S6 and 7 ).

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

Journal: Journal of Extracellular Vesicles

Article Title: Chemoradiotherapy‐Integrated Tumor Cell‐Derived Microparticles Mediate Tumor Eradication in Malignant Pleural Effusion

doi: 10.1002/jev2.70277

Figure Lengend Snippet: CR‐MPs promote APCs activation in vitro . (A) MFI of CRT on control, MTX treated, RT‐MPs treated and CR‐MPs@MTX treated LLC cells. (B, C) Relative (B) HMGB1 level and (C) ATP level in culture supernatant of control, MTX treated, RT‐MPs treated and CR‐MPs@MTX treated LLC cells. (D) Representative immunofluorescence images of CRT (red) on control, MTX treated, RT‐MPs treated and CR‐MPs@MTX treated H1299 cells. Scale bar = 20 µm. (E, F) MFI of (E) CD80 and (F) CD206 on BMDMs co‐cultured with control, MTX treated, RT‐MPs treated and CR‐MPs@MTX treated LLC cells. (G) Phagocytosis ratio of BMDMs to control, MTX treated, RT‐MPs treated and CR‐MPs@MTX treated LLC cells. (H) Uptake of PKH26‐labeled, LLC‐derived CR‐MPs@MTX by BMDCs was assessed by flow cytometry at 4, 12, and 24 h. (I, J) MFI of (I) CD80 and (J) CD86 on control BMDCs and BMDCs treated with MTX, RT‐MPs and CR‐MPs@MTX. (K) KEGG pathway enrichment analysis in CR‐MPs@MTX‐treated BMDCs versus control BMDCs. (L) GSEA of the interferon‐alpha (IFNα) response pathway in CR‐MPs@MTX‐treated BMDCs versus control BMDCs. (M) Western blot analysis of phospho‐TBK1, phospho‐p65, phospho‐IRF3, and actin in CR‐MPs@MTX treated or MTX treated BMDCs at 5‐, 10‐, 30‐, and 240‐minute post‐treatment. (N) RT‐qPCR evaluated expression of indicated genes in BMDCs after 24‐hour treatment with MTX, RT‐MPs, or CR‐MPs@MTX, compared with untreated control. All data are presented as mean ± s.d. One‐way ANOVA with Tukey's multiple comparisons test was performed. * P < 0.05, ** P < 0.01, and *** P < 0.001; ns, no significance.

Article Snippet: Mouse HMGB1 (EM0382, Finetest) and human HMGB1 (E‐EL‐H1554, Elabscience) produced in the supernatants were evaluated by ELISA assays according to the manufacturer's instructions.

Techniques: Activation Assay, In Vitro, Control, Immunofluorescence, Cell Culture, Labeling, Derivative Assay, Flow Cytometry, Western Blot, Quantitative RT-PCR, Expressing