Journal: Advanced Science

Article Title: A Time‐Programmed Bilayer Wound Dressing for Dynamic Microenvironment Modulation and Full‐Thickness Regeneration in Diabetic Wounds

doi: 10.1002/advs.202512425

Figure Lengend Snippet: Polyphenols scavenge AGEs and inflammatory factors. Gene set enrichment analysis (GSEA) of (A) defense response, (B) negative regulation of IFN‐I mediated signaling pathway, (C) negative regulation of IL‐6, and (D) activation of innate immune response. Differences in (E) AGEs, (F) RAGE, (G) IL‐4, (H) IL‐6, (I) IL‐10, and (L) TNF‐α levels between groups Control, T2DM, and Polyphenol were detected by Elisa. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. n = 6 independent samples for each group, error bars represent mean ± SD.

Article Snippet: Mouse TNF alpha PicoKine ELISA Kit (catalog no. EK0527), Mouse IL‐10 PicoKine ELISA Kit (catalog no. EK0417), Mouse IL‐6 PicoKine ELISA Kit (catalog no. EK0411), Mouse IL‐4 PicoKine ELISA Kit (catalog no. EK0405) were purchased from BOSTER (Wuhan, China).

Techniques: Activation Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: A Time‐Programmed Bilayer Wound Dressing for Dynamic Microenvironment Modulation and Full‐Thickness Regeneration in Diabetic Wounds

doi: 10.1002/advs.202512425

Figure Lengend Snippet: Expression of inflammatory and anti‐inflammatory factors during wound healing. (A) Representative images of immunohistochemical staining of IL‐4, (B) IL‐10, (C) TNF‐α, (D) IL‐6 in Control, Nanofiber, Polyhenol, PDGF‐BB, and Polyhenol+PDGF‐BB groups at 3 and 7 days. (E–H) Quantification of IL‐4, IL‐10, TNF‐α, and IL‐6. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Mouse TNF alpha PicoKine ELISA Kit (catalog no. EK0527), Mouse IL‐10 PicoKine ELISA Kit (catalog no. EK0417), Mouse IL‐6 PicoKine ELISA Kit (catalog no. EK0411), Mouse IL‐4 PicoKine ELISA Kit (catalog no. EK0405) were purchased from BOSTER (Wuhan, China).

Techniques: Expressing, Immunohistochemical staining, Staining, Control

Journal: Current Issues in Molecular Biology

Article Title: IL-37 Ameliorates Chronic Endometritis by Attenuating Epithelial—Mesenchymal Transition and Promoting M2 Macrophage Polarization

doi: 10.3390/cimb48020227

Figure Lengend Snippet: IL-37 regulates macrophage polarization and related signaling pathways in vitro (RAW 264.7 cells). ( A ) In the M1 polarization model, the relative mRNA expression levels of M1 macrophage-related marker genes (IL-6, iNOS, IL-1β) and signaling molecules (STAT3) were assessed by RT-qPCR. ( B ) In the M2 polarization model, the relative mRNA expression levels of M2 macrophage-related marker genes (TGF-β, CD206, Arg1) and signaling molecules (STAT6) were also evaluated by RT-qPCR. ( C ) (Left) Representative histogram of CD206 protein expression in macrophages detected by flow cytometry; (Right) Quantitative statistical graph of the percentage of CD206-positive cells. All data are presented as mean ± standard deviation; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The sections were then incubated overnight at 4 °C with primary antibodies against the following proteins: IL-37 (Abcam, Cambridge, UK), E-cadherin (Cell Signaling Technology [CST], Danvers, MA, USA), vimentin (ABclonal, Woburn, MA, USA), CD86 (Proteintech, Rosemont, IL, USA), CD206 (Proteintech), Smad3 (CST), and STAT6 (Proteintech).

Techniques: Protein-Protein interactions, In Vitro, Expressing, Marker, Quantitative RT-PCR, Flow Cytometry, Standard Deviation

Journal: Current Issues in Molecular Biology

Article Title: IL-37 Ameliorates Chronic Endometritis by Attenuating Epithelial—Mesenchymal Transition and Promoting M2 Macrophage Polarization

doi: 10.3390/cimb48020227

Figure Lengend Snippet: IL-37 regulates the activation, transcriptional activity, and nuclear translocation of the STAT6/Smad3 signaling pathway in macrophages. ( A ) Western blot analysis was performed to detect the protein expression bands of STAT6, phosphorylated STAT6 (p-STAT6), Smad3, phosphorylated Smad3 (p-Smad3), and arginase 1 (ARG1) in macrophages from different groups (RAW 264.7 cells). ( B ) A quantitative analysis graph of the grayscale values corresponding to the protein bands. ( C ) Dual luciferase reporter gene assays were conducted to investigate the effect of IL-37 on the transcriptional activity of the Smad pathway (HEK-293T cells). Data are expressed as the ratio of luciferase activity (firefly luciferase/Renilla luciferase). ( D ) (Left) Representative immunofluorescence images showing subcellular localization of STAT6 (green) and Smad3 (red) in RAW 264.7 cells after IL-37 treatment at 0 min, 30 min, and 1 h, with nuclei stained with DAPI (blue). Scale bar: 25 μm. (Right) Quantitative statistical results of the nuclear-to-cytoplasmic ratio of STAT6 and Smad3 at each time point. All data are expressed as mean ± standard deviation; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The sections were then incubated overnight at 4 °C with primary antibodies against the following proteins: IL-37 (Abcam, Cambridge, UK), E-cadherin (Cell Signaling Technology [CST], Danvers, MA, USA), vimentin (ABclonal, Woburn, MA, USA), CD86 (Proteintech, Rosemont, IL, USA), CD206 (Proteintech), Smad3 (CST), and STAT6 (Proteintech).

Techniques: Activation Assay, Activity Assay, Translocation Assay, Western Blot, Expressing, Luciferase, Immunofluorescence, Staining, Standard Deviation

Journal: The FASEB Journal

Article Title: Monosodium urate crystals alter the circadian clock in macrophages leading to loss of NLRP3 inflammasome repression: Implications for timing of the gout flare

doi: 10.1096/fj.202202035r

Figure Lengend Snippet: FIGURE 4 The effect of monosodium urate (MSU) crystals on Caspase 1 activity and IL-1β secretion is time-of-day dependent. (A) Schematic of the experimental design to assess the time-of-day differences in NLRP3 inflammasome activity. Following 18 h serum starvation, cells were media changed to serum-replete media and either treated immediately with MSU crystals (500 μg/mL) or rested for 12 h before media-changing again with/without MSU crystal addition. Caspase 1 activity was measured following 12 h of MSU crystal exposure and normalized for differences in cell number as determined by CyQuant assay with reference to a standard curve. (B) The magnitude of Caspase 1 activity induced by MSU crystal exposure in cells at the 0–12 h compared with 12–24 h timepoints. (C) The magnitude of increase in secreted IL-1β levels induced by MSU crystal exposure in THP-1 macrophages at the 0–12 h compared with 12–24 h timepoints. Data shown are mean ± SEM. Data were analyzed by t-test with p < .05 considered statistically significant.

Article Snippet: Secreted IL- 1β protein levels were measured in the cell culture supernatant using a Duoset Human IL- 1β ELISA kit (R&D Systems, Minneapolis, MN, USA) following the manufacturer's protocol.

Techniques: Activity Assay, CyQUANT Assay

Journal: The FASEB Journal

Article Title: Monosodium urate crystals alter the circadian clock in macrophages leading to loss of NLRP3 inflammasome repression: Implications for timing of the gout flare

doi: 10.1096/fj.202202035r

Figure Lengend Snippet: FIGURE 6 BMAL1 represses expression of pro-IL-1β and CASP1 but not NLRP3 in THP-1 macrophages BMAL1 was knocked down or over-expressed in THP-1 macrophages using adenoviral-mediated gene delivery. Following gene transduction, cells were serum starved for 18 h then media changed to serum-replete media and either treated immediately with MSU crystals (500 μg/mL) (0–12 h timepoint) or rested for 12 h before media-changing again with/without MSU crystal addition (12–24 h timepoint). NLRP3 expression measured by RT-qPCR following BMAL1 knockdown at (A) the 0–12 h timepoint and (B) 12–24 h timepoint. NLRP3 expression measured by RT-qPCR following BMAL1 overexpression at (C) the 0–12 h timepoint and (D) 12–24 h timepoint. Pro-IL-1β expression measured by RT-qPCR following BMAL1 knockdown at (E) the 0–12 h timepoint and (F) 12–24 h timepoint. Pro-IL-1β expression measured by RT-qPCR following BMAL1 overexpression at (G) the 0–12 h timepoint and (H) 12–24 h timepoint. CASP1 expression measured by RT-qPCR following BMAL1 knockdown at (I) the 0–12 h timepoint and (J) 12–24 h timepoint. CASP1 expression measured by RT-qPCR following BMAL1 overexpression at (K) the 0–12 h timepoint and (L) 12–24 h timepoint. Data shown are mean ± SEM for 3 experimental replicates. Data were analyzed by one-way ANOVA (post-hoc Tukey) with p < .05 considered statistically significant.

Article Snippet: Secreted IL- 1β protein levels were measured in the cell culture supernatant using a Duoset Human IL- 1β ELISA kit (R&D Systems, Minneapolis, MN, USA) following the manufacturer's protocol.

Techniques: Expressing, Transduction, Quantitative RT-PCR, Knockdown, Over Expression

Journal: The FASEB Journal

Article Title: Monosodium urate crystals alter the circadian clock in macrophages leading to loss of NLRP3 inflammasome repression: Implications for timing of the gout flare

doi: 10.1096/fj.202202035r

Figure Lengend Snippet: FIGURE 7 REV-ERBα represses NLRP3 expression and inflammasome activity in THP-1 macrophages. Following 18 h serum starvation, cells were re-fed with serum-replete media and either treated immediately with/without MSU crystals (500 μg/mL) and heme (30 μM)), SR8278 (1 μM), heme+SR8278 or vehicle (0.1M NaOH + 0.1% DMSO) for 12 h (0–12 h timepoint) or rested for 12 h and then media-changed to fresh serum-replete media with/without MSU crystals and heme/SR8278/vehicle (12–24 h timepoint). (A) Caspase-1 activity normalized to cell number as measured at the 0–12 h timepoint and (B) 12–24 h timepoint. (C) Levels of secreted IL-1β measured by ELISA at the 0–12 h and (D) 12–24 h timepoint. To ensure all cells were exposed to the same vehicles, SR8278 vehicle (0.1%) DMSO) was added to the heme-only treatment and heme vehicle (0.1M NaOH) added to the SR8278-only treatment. Data shown are mean ± SEM for 3 experimental replicates. All data were analyzed by one-way ANOVA (post-hoc Tukey) p < .05 was considered statistically significant.

Article Snippet: Secreted IL- 1β protein levels were measured in the cell culture supernatant using a Duoset Human IL- 1β ELISA kit (R&D Systems, Minneapolis, MN, USA) following the manufacturer's protocol.

Techniques: Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay