Journal: Experimental and therapeutic medicine

Article Title: Blood purification treatment initiated at the time of sepsis diagnosis effectively attenuates serum HMGB1 upregulation and improves patient prognosis.

doi: 10.3892/etm.2017.4854

Figure Lengend Snippet: Figure 1. Serum and urine HMGB1 levels upon diagnosis of sepsis. (A) Serum and (B) urine levels of HMGB1 were measured by ELISA in the different patient groups prior to the initiation of treatment. Data are presented as the mean ± standard deviation. #P<0.05 vs. Control. HMGB1, high mobility group box protein 1; CRRT, continuous renal replacement treatment; HP, hemoperfusion.

Article Snippet: Serum and urine HMGB1 levels were measured using a Human HMGB1 ELISA kit (E-EL-H1554c, Elabscience Biotechnology Co., Ltd., Wuhan, China) according to the manufacturer's protocol.

Techniques: Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Standard Deviation, Control

Journal: Experimental and therapeutic medicine

Article Title: Blood purification treatment initiated at the time of sepsis diagnosis effectively attenuates serum HMGB1 upregulation and improves patient prognosis.

doi: 10.3892/etm.2017.4854

Figure Lengend Snippet: Figure 2. Changes in serum and urine HMGB1 levels over 24 h. (A) Serum and (B) urine levels of HMGB1 in the sepsis (no apheresis intervention) group at different time points. Data are presented as the mean ± standard deviation. #P<0.05 vs. 0‑h value. HMGB1, high mobility group box protein 1.

Article Snippet: Serum and urine HMGB1 levels were measured using a Human HMGB1 ELISA kit (E-EL-H1554c, Elabscience Biotechnology Co., Ltd., Wuhan, China) according to the manufacturer's protocol.

Techniques: Standard Deviation

Journal: Experimental and therapeutic medicine

Article Title: Blood purification treatment initiated at the time of sepsis diagnosis effectively attenuates serum HMGB1 upregulation and improves patient prognosis.

doi: 10.3892/etm.2017.4854

Figure Lengend Snippet: Figure 3. Association of serum HMGB1 level with urine HMGB1 level and APACHE II score. Correlations were identified between (A) urine and serum HMGB1 levels, and (B) APACHE II score and serum HMGB1 level in the sepsis (no apheresis intervention) group. HMGB1, high mobility group box protein 1; APACHE II, Acute Physiology and Chronic Health Evaluation II.

Article Snippet: Serum and urine HMGB1 levels were measured using a Human HMGB1 ELISA kit (E-EL-H1554c, Elabscience Biotechnology Co., Ltd., Wuhan, China) according to the manufacturer's protocol.

Techniques:

Journal: Experimental and therapeutic medicine

Article Title: Blood purification treatment initiated at the time of sepsis diagnosis effectively attenuates serum HMGB1 upregulation and improves patient prognosis.

doi: 10.3892/etm.2017.4854

Figure Lengend Snippet: Figure 4. Changes in serum and urine HMGB1 levels in the different patient groups over 24 h. (A) Serum and (B) urine levels of HMGB1 were measured by ELISA at different time points after the initiation of treatments. Data are presented as the mean ± standard deviation. #P<0.05 vs. 0‑h value; *P<0.05 vs. sepsis group; ^P<0.05 vs. 12‑h value. HMGB1, high mobility group box protein 1; CRRT, continuous renal replacement treatment; HP, hemoperfusion.

Article Snippet: Serum and urine HMGB1 levels were measured using a Human HMGB1 ELISA kit (E-EL-H1554c, Elabscience Biotechnology Co., Ltd., Wuhan, China) according to the manufacturer's protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: OncoImmunology

Article Title: Carbon ion radiotherapy triggers immunogenic cell death and sensitizes melanoma to anti-PD-1 therapy in mice

doi: 10.1080/2162402x.2022.2057892

Figure Lengend Snippet: Figure 1. CIRT-induced immunogenic cell death in vitro. Human osteosarcoma U2OS cells were irradiated by X-ray and carbon ion radiotherapy (CIRT), and biomarkers of immunogenic cell death were evaluated at 6 h and 24 h after irradiation. Mitoxantrone (MTX, 1 µM) was used as a prototype immunogenic cell death inducer Flow cytometry, immunofluorescence, and immunoblot assays showed that CIRT induced CALR exposure and elevated the phosphorylation level of eIF2α, enhanced ATP release by quinacrine staining and ELISA, augmented HMGB1 exodus from the nucleus, and significantly up-regulated the expression of type I interferon at the mRNA level. Representative images and quantification are shown (mean ± SD of triplicate assessments, Student’s t test, **p < .01, ***p < .001).

Article Snippet: The HMGB1 (#E-EL-M0676c) and IFN-γ (#E-EL-M0048c) ELISA kits were purchased from Elabscience (Wuhan, China); the ENLITEN ATP assay kits (#ff2000) was purchased from Promega (Madison, WI, USA).

Techniques: In Vitro, Irradiation, Flow Cytometry, Immunofluorescence, Western Blot, Phospho-proteomics, Staining, Enzyme-linked Immunosorbent Assay, Expressing

Journal: OncoImmunology

Article Title: Carbon ion radiotherapy triggers immunogenic cell death and sensitizes melanoma to anti-PD-1 therapy in mice

doi: 10.1080/2162402x.2022.2057892

Figure Lengend Snippet: Figure 2. CIRT-induced immunogenic cell death in vivo. 5 × 105 B16 and S91 cells were irradiated with 5 GyE of carbon ion beams or treated with 2 μM MTX for 24 h respectively, then subcutaneously inoculated in C57BL/6 mice. 2 weeks after, rechallenged with 5 × 104 B16 and S91 cells and the tumor growth was documented regularly. C57BL/6 mice bearing subcutaneous B16 or S91 melanoma were locally irradiated with X-rays (XR, 5 Gy) or carbon ions (CIRT, 5 GyE) at the tumor site. Mitoxantrone (MTX, 2.0 mg/Kg)-treated animals were used as positive control. Eight days after irradiation, CALR exposure, eIF2α phosphorylation, and HMGB1 exodus were examined by immunofluorescence. The secretion of HMGB1, ATP, and IFN-γ in the serum was determined by ELISA; and infiltration of IFN-γ expressing cell in the tumor bed was assessed by immunohistocheistry. Representative images and quantification are shown (mean ± SD of triplicate assessments, Student’s t test, ***p < .001).

Article Snippet: The HMGB1 (#E-EL-M0676c) and IFN-γ (#E-EL-M0048c) ELISA kits were purchased from Elabscience (Wuhan, China); the ENLITEN ATP assay kits (#ff2000) was purchased from Promega (Madison, WI, USA).

Techniques: In Vivo, Irradiation, Positive Control, Phospho-proteomics, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Expressing

Journal: OncoImmunology

Article Title: Carbon ion radiotherapy triggers immunogenic cell death and sensitizes melanoma to anti-PD-1 therapy in mice

doi: 10.1080/2162402x.2022.2057892

Figure Lengend Snippet: Figure 3. Combination therapy with CIRT and anti-PD-1 stimulates immunogenic cell death. C57BL/6 mice bearing subcutaneous B16 and S91 melanoma (Model) were injected intraperitoneally with anti-PD-1 (α-PD-1) or locally irradiated with 5 GyE of carbon ions (CIRT) at tumor sites, or received treatment with 5 Gy CIRT plus anti-PD-1 (CIRT+α-PD-1). Eight days after irradiation, we examined CALR exposure, eIF2α phosphorylation, and HMGB1 exodus by immunofluorescence; secretion of IFN-γ in serum by ELISA; and IFN-γ infiltration in the tumor by immunohistochemical assay. Representative images and quantification are shown (mean ± SD of triplicate assessments, Student’s t test, ***p < .001).

Article Snippet: The HMGB1 (#E-EL-M0676c) and IFN-γ (#E-EL-M0048c) ELISA kits were purchased from Elabscience (Wuhan, China); the ENLITEN ATP assay kits (#ff2000) was purchased from Promega (Madison, WI, USA).

Techniques: Injection, Irradiation, Phospho-proteomics, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining

Journal: International immunopharmacology

Article Title: NRICM101 prevents kainic acid-induced seizures in rats by modulating neuroinflammation and the glutamatergic system.

doi: 10.1016/j.intimp.2024.112842

Figure Lengend Snippet: Fig. 8. Effect of NRICM101 on HMGB1 and TLR4 proteins in the cortex and hippocampus. (A) Schematic representation of HMGB1/TLR4 signaling pathway. (B, C) Immunoblot results of HMGB1 and TLR4 proteins in the cortex and hippocampus from different groups and the respective bar graphs. (D) HMGB1 in serum in each group detected with ELISA. Data were expressed as mean ± SEM. One-way ANOVA with Tukey posthoc test. *** significant vs control p < 0.001, # significant vs KA group p < 0.05. n = 5 rats per group.

Article Snippet: The serum levels of HMGB1, AST and ALT in rats were detected using ELISA kits (HMGB1, Novus, #NBP3-06661; ALT, Abcam, #ab234579; AST, Abcam, #ab263883) with a microplate ELISA reader.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: International immunopharmacology

Article Title: NRICM101 prevents kainic acid-induced seizures in rats by modulating neuroinflammation and the glutamatergic system.

doi: 10.1016/j.intimp.2024.112842

Figure Lengend Snippet: Fig. 10. Effect of NRICM101 on body weight, liver, kidney, and serum ALT and AST of rats. (A) Graphical representation of animal body weight changes relative to the initial body weight. (B) Photomicrographs of rat’s liver and kidneys following H&E staining (200 × ). Scale bars = 50 μm. (C) ALT and AST in serum detected with ELISA. Data were expressed as mean ± SEM. One-way ANOVA with Tukey posthoc test. n = 8–13 rats per group.

Article Snippet: The serum levels of HMGB1, AST and ALT in rats were detected using ELISA kits (HMGB1, Novus, #NBP3-06661; ALT, Abcam, #ab234579; AST, Abcam, #ab263883) with a microplate ELISA reader.

Techniques: Staining, Enzyme-linked Immunosorbent Assay

Journal: Nature Communications

Article Title: Development of a RIPK1 degrader to enhance antitumor immunity

doi: 10.1038/s41467-024-55006-2

Figure Lengend Snippet: B16F10 cells were treated with or without TNFα (100 ng/mL), Z-VAD-FMK (25 μM), LD4172 (1 μM), and/or T2I (1 μM) for 72 h. A NF-κB activity in B16F10 cells expressing a NanoLuc reporter for NF-κB response. Data represent the mean NF-κB activity ± SD ( n = 3 biologically independent samples from two independent experiments). Statistical significance was determined using a two-tailed unpaired t -test, with P values indicated. B Representative flow cytometry dot plots showing apoptosis in B16F10 cells from three independent experiments ( n = 3 biologically independent samples per experiment). Viable cells (FITC−/PI−) are located in the lower left quadrant, early apoptotic cells (FITC+/PI−) in the lower right quadrant, and late apoptotic cells (FITC+/PI+) in the upper right quadrant. C Representative images of B16F10 cells stained with PI (red) and caspase 3/7 (green). Data are shown from three independent experiments ( n = 3 biologically independent samples per experiment). Cell death was quantified by PI uptake using the Cytation 5 imager. D Top: Percentage of PI+ B16F10 cells after 72 h of the indicated treatments, calculated as PI+ (%) = (Number of PI+ cells/Total cells) × 100%, using the Cytation 5 imager. Data represent the mean ± SD ( n = 6 biologically independent replicates from three independent experiments). Statistical significance was determined using a two-tailed unpaired t -test, with P values indicated. Bottom: Western blots showing expression of cleaved caspase-3, cleaved caspase-7, and cleaved PARP in B16F10 cells from one experiment. Source data are provided as a file. E Top: Quantification of extracellular ATP levels secreted by B16F10 cells. Data represent mean extracellular ATP levels ± SD ( n = 4 biologically independent samples from one experiment), with statistical significance determined by a two-tailed unpaired t -test, and P values indicated. Bottom: Representative Western blots showing expression of HMGB1 and calreticulin in B16F10 cells. Source data are provided as a file. F Immunofluorescence staining of HMGB1 in B16F10 cells, with data from three biologically independent samples from one experiment.

Article Snippet: Serum HMGB1 was analyzed by ELISA (NOVUS, NBP2-62767) according to the manufacturer’s protocol.

Techniques: Activity Assay, Expressing, Two Tailed Test, Flow Cytometry, Staining, Western Blot, Immunofluorescence

Journal: Nature Communications

Article Title: Development of a RIPK1 degrader to enhance antitumor immunity

doi: 10.1038/s41467-024-55006-2

Figure Lengend Snippet: A Representative immunofluorescent images showing RIPK1, Calreticulin, CD8, CD4, Foxp3, and F4/80 staining, along with hematoxylin and eosin (H&E) staining and cleaved caspase-3/7 levels in B16F10 tumors following 5 days of the indicated treatment. Images represent four independent fields per slide, with one slide from each of four mice per group. Scale bars: 20 μm and 200 μm (both at 60× objective). B The mouse plasma HMGB1 level from different treatment groups ( n = 8 mice). Statistical significance was calculated with a two-tailed unpaired t -test and P values are indicated. Flow cytometry quantification of PD1 + CD8+ T cells C CD4+ T cells D cDC cells E macrophages F and CD8+ T cells G in B16F10 tumors following 5 days of indicated treatment ( n = 10 mice/group). H Flow cytometric quantification of IFNγ+ CD8+ T cells in B16F10 tumors treated with the indicated treatments for 5 days and stimulated with PMA/ionomycin in vitro for 6 h ( n = 5 tumors/group). Statistical significance was calculated with a two-tailed unpaired t -test and P values are indicated. I Tumor volume (mm 3 ) of vehicle-, anti-PD1−, LD4172−, anti-PD1+LD4172−, and anti-PD1+LD4172+anti-CD8–treated B16F10 tumors ( n = 8 mice). Each symbol represents the mean tumor volume, with error bars indicating SEM. Statistical analysis was performed using two-way ANOVA followed by Sidak’s multiple comparisons test, with significance levels indicated. J Heat map showing log 10-fold changes in the concentration of mouse plasma cytokines normalized by the mean value of control mice ( n = 8 mice). For all experiments: LD4172: 20 mg /kg; anti-PD1 antibody: 100 μg/mice; anti-CD8: 100 μg/mice; Combo: LD4172 plus anti-PD1.

Article Snippet: Serum HMGB1 was analyzed by ELISA (NOVUS, NBP2-62767) according to the manufacturer’s protocol.

Techniques: Staining, Clinical Proteomics, Two Tailed Test, Flow Cytometry, In Vitro, Concentration Assay, Control