Journal: Materials today. Bio

Article Title: Precise delivery of doxorubicin and imiquimod through pH-responsive tumor microenvironment-active targeting micelles for chemo- and immunotherapy.

doi: 10.1016/j.mtbio.2022.100482

Figure Lengend Snippet: Fig. 10. Immunostaining of tumor tissues in tumor-bearing mice after treatments. (A) Immunohistochemistry images of tumor sections stained with CD3, CD8, and TNF-α antibodies. The scale bar is 100 μm. (B) Immunofluorescence images of tumor tissues after treatments for 12 days. The scale bar is 50 μm. Blue fluorescence represents the cell nucleus stained with DAPI. Green fluorescence represents the iNOS stained with the iNOS antibody conjugated with FITC.

Article Snippet: After 30 min, the tissue slice was stained with diluted iNOS antibody (Miltenyi Biotec, catalog: 130-116-357) at 4 C overnight.

Techniques: Immunostaining, Immunohistochemistry, Staining

Journal: Pharmaceuticals

Article Title: Anti-Inflammatory and Antioxidant Effects of Topical Formulations Containing Plant Extracts, Methylsulfonylmethane, and Peptiskin® in In Vitro Models of Arthritis

doi: 10.3390/ph18091270

Figure Lengend Snippet: Figure 8. Protective effect of AS632 against TNF-α/IL-1β-induced inflammatory responses in C28/I2 chondrocytes. C28/I2 chondrocytes were pre-treated with empty liposomes (ELs), AS632, or AS633 at a concentration of 1 µL/mL or 1 µM dexamethasone as a positive control for 2 h, followed by stimulation with 1 ng/mL IL-1β and 10 ng/mL TNF-α. (A) After 6 h of treatment, total RNA was extracted, and the mRNA expression levels of IL-6, TNF-α, and IL-1β were analyzed by real-time qPCR. (B) After 24 h of treatment, cells were lysed, and protein levels of MMP-3, MMP-13, and iNOS were analyzed by Western blotting. * p < 0.05; ** p < 0.01 compared to the cytokine-treated control group.

Article Snippet: Antibodies against iNOS (Cat. No. sc-7271) and MMP-9 (Cat. No. sc-13520) were acquired from Santa Cruz Biotechnology (Dallas, TX, USA), while the MMP-3 antibody (Cat. No. LS-C27030-200) was obtained from LS Bio (Seattle, WA, USA).

Techniques: Liposomes, Concentration Assay, Positive Control, Expressing, Western Blot, Control

Journal: Materials Today Bio

Article Title: Neuro-bone-skin tri-regeneration via a microenvironment-responsive PRP-loaded chitosan hydrogel for traumatic brain injury therapy

doi: 10.1016/j.mtbio.2026.102913

Figure Lengend Snippet: CO@P hydrogel suppresses neuroinflammation. A-B) iNOS and Arg1 immunofluorescence: Day 3 vs. Day 28. Scale bar = 100 μm. C) Microglial polarization shift schematic (By Figdraw). D-E) Quantified fluorescence intensity of iNOS and Arg1 in each group. F-G) Quantitative analysis of the ELISA results of IL4, IL10, TNF-α and IL1-β on Day 3 and Day 28. Data were presented as mean ± SEM. N = 3 per group. ∗: P < 0.05 compared with Sham group; #: P < 0.05 compared with TBI group; Δ: P < 0.05 compared with CO@P group; .

Article Snippet: Primary antibodies used included NeuN (1100 μg/mL), GFAP (800 μg/mL), Iba1 (700 μg/mL), iNOS (820 μg/mL), Arg1 (900 μg/mL), VEGF (1000 μg/mL), CD31 (600 μg/mL), Caspase-3 (700 μg/mL), DCX (600 μg/mL) (1:200 dilution; Proteintech, China).

Techniques: Immunofluorescence, Fluorescence, Enzyme-linked Immunosorbent Assay

Journal: Oncotarget

Article Title: Subanesthetic isoflurane relieves zymosan-induced neutrophil inflammatory response by targeting NMDA glutamate receptor and Toll-like receptor 2 signaling

doi: 10.18632/oncotarget.9091

Figure Lengend Snippet: A. Neutrophils were treated with increasing concentrations of zymosan for indicated time periods, and were subjected to Western blot analysis. B. Neutrophils were incubated with zymosan for 15 min, post-treated with 0.7% isoflurane for 15 min, followed by a continuous incubation with zymosan for a total of 8 h before Western blot analysis. β-actin was used as an inner control for whole cell lysates. C. Neutrophils were co-transfected with or without an iNOS-luc reporter plasmid and indicated siRNAs. The promoter activity of iNOS was determined in the cell lysates after zymosan treatment for 6 h. D. , E. Neutrophils were pretreated with NAI (D) or transfected with NF-κB p65 siRNA (E), followed by incubation with zymosan for 8 h. The nuclear or whole cell lysates were prepared for Western blot analysis. Lamin B was used as an inner control for the lysates of the nuclear fraction. F. Neutrophils were treated as described in (D), followed by incubation with zymosan for 6 h prior to RT-PCR analysis. G. Neutrophils were treated as described in (D), followed by measurement of NO and ONOO − production. H. Neutrophils were treated as described in (B), and the nuclear lysates were prepared for Western blot analysis. Data are represented as the mean ± SEM of 3 replicates or representative of 3 independent experiments. * P < 0.05, # P < 0.01, as compared with the cells exposed to vehicle alone (A). * P < 0.05, # P < 0.01, as compared with the cells exposed to zymosan alone (B-H), or zymosan + scrambled siRNA (E).

Article Snippet: The small interfering RNA (siRNA) duplexes corresponding to mouse TLR2 (sc-40257), TLR4 (sc-40261), MyD88 (sc-35987), c-Src (sc-29859), p47 phox (sc-151963), p38 MAPK (sc-29434), NF-κB p65 (sc-29411), iNOS (sc-36092), NMDA receptor subunit NR1 (sc-36082), and scrambled siRNA (sc-44230) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, Texas, USA).

Techniques: Western Blot, Incubation, Control, Transfection, Plasmid Preparation, Activity Assay, Reverse Transcription Polymerase Chain Reaction

Journal: Oncotarget

Article Title: Subanesthetic isoflurane relieves zymosan-induced neutrophil inflammatory response by targeting NMDA glutamate receptor and Toll-like receptor 2 signaling

doi: 10.18632/oncotarget.9091

Figure Lengend Snippet: Zymosan induces ROS production through a TLR2/MyD88/c-Src/NADPH oxidase pathway, which in turn caused the activation of p38 MAPK and consequently the activation and nuclear translocation of NF-κB. NF-κB thus switches on iNOS expression, and increases NO production and ONOO − release from neutrophils, which eventually permeabilizes adjacent PMVECs. Subanesthetic isoflurane exerts a protective role for this pathological process by inhibiting c-Src activity, which is probably attributed to isoflurane targeting of the NMDA glutamate receptor and thereby suppression of the calcium signaling in neutrophils.

Article Snippet: The small interfering RNA (siRNA) duplexes corresponding to mouse TLR2 (sc-40257), TLR4 (sc-40261), MyD88 (sc-35987), c-Src (sc-29859), p47 phox (sc-151963), p38 MAPK (sc-29434), NF-κB p65 (sc-29411), iNOS (sc-36092), NMDA receptor subunit NR1 (sc-36082), and scrambled siRNA (sc-44230) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, Texas, USA).

Techniques: Activation Assay, Translocation Assay, Expressing, Activity Assay