Journal: Pain medicine (Malden, Mass.)

Article Title: Neonatal bee venom exposure induces sensory modality-specific enhancement of nociceptive response in adult rats.

doi: 10.1111/pme.12296

Figure Lengend Snippet: Figure 1 A flowchart illustrating the experimental design.

Article Snippet: After being blocked in 5% normal goat serum in PBS containing 0.3% Triton X-100 for 30 minutes, sections were incubated overnight at 4°C with primary antibodies to NGF (1:200; rabbit anti-rat, Santa Cruz Biotechnology, Santa Cruz, CA, USA), TrkA receptor (1:200; rabbit antirat, Santa Cruz), BDNF (1:200; rabbit anti-rat, Santa Cruz), TrkB receptor (1:200; rabbit anti-rat, Santa Cruz), IL-1β (1:200; rabbit anti-rat, Santa Cruz), or COX-2 (1:200; goat anti-rat, Santa Cruz).

Techniques:

Journal: Pain medicine (Malden, Mass.)

Article Title: Neonatal bee venom exposure induces sensory modality-specific enhancement of nociceptive response in adult rats.

doi: 10.1111/pme.12296

Figure Lengend Snippet: Figure 8 Effects of neonatal inflammation on the spinal expression of IL-1β and COX-2 in adult rats. Compared with adult rats without neonatal inflammation, rats receiving the first BV injection on P21 and beyond showed up-regulated expression of IL-1β in the spinal cord dorsal horn. Scale bar: 200 μm. IOD = integral optical density. Sal-BV = rats receiving saline injection neonatally and then exposed to BV as adults. BV-BV = rats receiving BV neonatally and then again exposed to BV as adults. * P < 0.05, as compared with the Sal-BV group.

Article Snippet: After being blocked in 5% normal goat serum in PBS containing 0.3% Triton X-100 for 30 minutes, sections were incubated overnight at 4°C with primary antibodies to NGF (1:200; rabbit anti-rat, Santa Cruz Biotechnology, Santa Cruz, CA, USA), TrkA receptor (1:200; rabbit antirat, Santa Cruz), BDNF (1:200; rabbit anti-rat, Santa Cruz), TrkB receptor (1:200; rabbit anti-rat, Santa Cruz), IL-1β (1:200; rabbit anti-rat, Santa Cruz), or COX-2 (1:200; goat anti-rat, Santa Cruz).

Techniques: Expressing, Injection, Saline

Journal: Cell Death & Disease

Article Title: LincRNA-Gm4419 knockdown ameliorates NF- κ B/NLRP3 inflammasome-mediated inflammation in diabetic nephropathy

doi: 10.1038/cddis.2016.451

Figure Lengend Snippet: Gm4419 regulates inflammation, fibrosis and proliferation of MCs under high-glucose condition. ( a ) The mRNA levels of pro-inflammatory cytokines (mcp-1, TNF- α and IL-1 β ) and fibrosis biomarkers (Fn and Col.IV) in MCs that were stimulated by high or low glucose by qRT-PCR analysis. ( b ) The mRNA levels of pro-inflammatory cytokines and fibrosis biomarkers by qRT-PCR analysis in cells overexpressed or downexpressed Gm4419. ( c ) The protein levels of pro-inflammatory cytokines and fibrosis biomarkers in untransfected or transfected MCs by western blot and quantitative analysis. ( d ) The expressions of pro-inflammatory cytokines and fibrosis biomarkers in untransfected or transfected MCs by immunofluorescent (× 400). ( e ) Proliferative capability of untransfecting or transfecting MCs was analyzed by EdU (5-ethynyl-2′-deoxyuridine) incorporation assay, and the EdU incorporation rate was shown as the ratio of EdU-positive cells to total Hoechst33342-positive cells. ( f ) Untransfected or transfected MCs were analyzed by flow cytometry and quantitative analysis. The percentage of cells in the G0/G1, S and G2/M phases of the cell cycle were calculated. The data are representative of the results of three independent experiments, and the data were presented as means±S.E.M. (* P <0.05, ** P <0.01, NS, no significant)

Article Snippet: Both non-transfected and transfected MCs were fixed with 4% paraformaldehyde for 30 min at room temperature and then permeabilized in PBS containing 0.1% Triton X-100 on ice for 10 min. And then samples were confined by 3% goat serum (Beyotime, Nantong, China) for 1 h at room temperature and incubated by overnight at 4 °C using anti-p50 antibody (Abcam, 1:100), anti-p65 antibody (Abcam, 1:100), anti-NLRP3 inflammasome antibody (Sangon Bio Tech, 1:50), anti-mcp-1 antibody (Sangon Bio Tech, 1:50), anti-TNF- α antibody (Bioworld Tech, Minnesote, CA, USA, 1:50), anti-IL-1 β antibody (Bioworld Tech, 1:50), anti-Fn antibody (Sangon Bio Tech, 1:50) or anti-Col4 antibody (Proteintech, Wuhan, China, 1:50).

Techniques: Quantitative RT-PCR, Transfection, Western Blot, Flow Cytometry

Journal: Journal of Cellular and Molecular Medicine

Article Title: Phloretin attenuates hyperuricemia‐induced endothelial dysfunction through co‐inhibiting inflammation and GLUT 9‐mediated uric acid uptake

doi: 10.1111/jcmm.13176

Figure Lengend Snippet: Effects of UA on cell viability and inflammation in HUVEC s. ( A ) Cell viability of HUVEC s assayed by CCK 8. ( B‐C ) Real‐time PCR analysis of MCP ‐1 , VCAM ‐1 , ICAM ‐1 and IL ‐1 β mRNA . ( D ) Western blot of ICAM ‐1, IL ‐1β and MCP ‐1 protein, cell treated with UA for 24h. ( E ) Quantification analysis of protein level related to control (* P < 0.05, UA group versus control). HUVECs: human umbilical vein endothelial cells, NFκB/ERK: nuclear factor‐kappa B/extracellular regulated protein kinases.

Article Snippet: The membranes were blocked with 5% non‐fat milk and incubated with primary antibodies against p‐NFκB (1:1000, Abcam, MA, USA) and NFκB (1:500, Santa Cruz Biotechnology, CA, USA), GLUT9 (1:1000, Abcam), IL‐1β (1:500, ABclonal Biotechnology), OAT1 (1:1000, ABclonal ), URAT1 (1:1000,ABclonal) ICAM‐1 (1:1000, Cell Signaling Technology, MA, USA), MCP‐1 (1:1000, Abcam), VCAM‐1 (1:500, Proteintech, USA) overnight at 4°C.

Techniques: CCK-8 Assay, Real-time Polymerase Chain Reaction, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Phloretin attenuates hyperuricemia‐induced endothelial dysfunction through co‐inhibiting inflammation and GLUT 9‐mediated uric acid uptake

doi: 10.1111/jcmm.13176

Figure Lengend Snippet: Effect of phloretin on UA ‐induced inflammation in HUVEC s. ( A ) Cell viability of HUVEC s assayed by CCK 8, cell treated with phloretin for 24h. ( B‐C ) Real‐time PCR analysis of MCP ‐1 , VCAM ‐1 , ICAM ‐1 and IL ‐1 β mRNA , and cell treated with UA with or without phloretin for 24h. ( D ) Western blot and ( E ) quantification analysis of ICAM ‐1, IL ‐1β and MCP ‐1 protein (* P < 0.05, UA group versus control, # P < 0.05, phloretin group versus UA group). HUVECs: human umbilical vein endothelial cells, NFκB/ERK: nuclear factor‐kappa B/extracellular regulated protein kinases.

Article Snippet: The membranes were blocked with 5% non‐fat milk and incubated with primary antibodies against p‐NFκB (1:1000, Abcam, MA, USA) and NFκB (1:500, Santa Cruz Biotechnology, CA, USA), GLUT9 (1:1000, Abcam), IL‐1β (1:500, ABclonal Biotechnology), OAT1 (1:1000, ABclonal ), URAT1 (1:1000,ABclonal) ICAM‐1 (1:1000, Cell Signaling Technology, MA, USA), MCP‐1 (1:1000, Abcam), VCAM‐1 (1:500, Proteintech, USA) overnight at 4°C.

Techniques: CCK-8 Assay, Real-time Polymerase Chain Reaction, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Phloretin attenuates hyperuricemia‐induced endothelial dysfunction through co‐inhibiting inflammation and GLUT 9‐mediated uric acid uptake

doi: 10.1111/jcmm.13176

Figure Lengend Snippet: Effect of phloretin on UA / TNF ‐α‐induced inflammation and NF κB/ ERK activation. ( A‐B ) Western blot and quantification analysis of ICAM ‐1, IL ‐1β and MCP ‐1 protein, cell treated with TNF ‐α and phloretin. ( C‐D ) Western blot and quantification analysis of p‐ ERK / ERK and p‐ NF κB p65/ NF κB p65 levels. ( E ) Immunofluorescent staining and ( F ) quantification results of NF κB p65 nuclear translocation in HUVEC s (scale bar = 20 μm) (* P < 0.05, UA or TNF group versus control, # P < 0.05, phloretin group versus UA or TNF group). HUVECs: human umbilical vein endothelial cells, NFκB/ERK: nuclear factor‐kappa B/extracellular regulated protein kinases.

Article Snippet: The membranes were blocked with 5% non‐fat milk and incubated with primary antibodies against p‐NFκB (1:1000, Abcam, MA, USA) and NFκB (1:500, Santa Cruz Biotechnology, CA, USA), GLUT9 (1:1000, Abcam), IL‐1β (1:500, ABclonal Biotechnology), OAT1 (1:1000, ABclonal ), URAT1 (1:1000,ABclonal) ICAM‐1 (1:1000, Cell Signaling Technology, MA, USA), MCP‐1 (1:1000, Abcam), VCAM‐1 (1:500, Proteintech, USA) overnight at 4°C.

Techniques: Activation Assay, Western Blot, Staining, Translocation Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Phloretin attenuates hyperuricemia‐induced endothelial dysfunction through co‐inhibiting inflammation and GLUT 9‐mediated uric acid uptake

doi: 10.1111/jcmm.13176

Figure Lengend Snippet: Effect of GLUT 9 inhibition on UA uptake and inflammatory response in HUVEC s. ( A ) Western blot analysis of GLUT 9 expression in si RNA treated HUVEC s (24 hrs). ( B‐C ) Western blot analysis of SLC 2A9 si RNA on ICAM ‐1, IL ‐1β and NF κB/ ERK activation in UA ‐treated HUVEC s (* P < 0.05, UA group versus control, # P < 0.05, si RNA group versus UA group). HUVECs: human umbilical vein endothelial cells, NFκB/ERK: nuclear factor‐kappa B/extracellular regulated protein kinases.

Article Snippet: The membranes were blocked with 5% non‐fat milk and incubated with primary antibodies against p‐NFκB (1:1000, Abcam, MA, USA) and NFκB (1:500, Santa Cruz Biotechnology, CA, USA), GLUT9 (1:1000, Abcam), IL‐1β (1:500, ABclonal Biotechnology), OAT1 (1:1000, ABclonal ), URAT1 (1:1000,ABclonal) ICAM‐1 (1:1000, Cell Signaling Technology, MA, USA), MCP‐1 (1:1000, Abcam), VCAM‐1 (1:500, Proteintech, USA) overnight at 4°C.

Techniques: Inhibition, Western Blot, Expressing, Activation Assay