Journal: Phytomedicine

Article Title: Qingke Pingchuan granules alleviate airway inflammation in COPD exacerbation by inhibiting neutrophil extracellular traps in mice

doi: 10.1016/j.phymed.2024.156283

Figure Lengend Snippet: Fig. 7. Validation of compound-target interactions through molecular docking and protein-small molecule binding assays. (A) Chemical structure of EGCG and molecular docking models showing its interactions with NOX2 and p47phox. (B) Chemical structure of quercetin and molecular docking models illustrating its interactions with NOX2 and p47phox. (C) An SPR sensorgram displaying the binding affinity of the NOX2-EGCG interaction. (D) An SPR sensorgram showing the binding affinity of the NOX2-quercetin interaction. (E) A BLI sensorgram illustrating the binding affinity of the p47phox-EGCG interaction. (F)A BLI sensorgram depicting the binding affinity of the p47phox-quercetin interaction. Abbreviations: BLI, Bio-layer interferometry; EGCG, epigallocatechin-3-gallate; NOX2, NADPH oxidase 2; p47phox (NCF1), 47 kDa neutrophil oxidase factor; Que, quercetin; SPR, surface plasmon resonance.

Article Snippet: Recombinant human NOX2 protein was acquired from CUSABIO (Wuhan, China), while recombinant human P47phox protein was sourced from TargetMol Co., Ltd. (USA).

Techniques: Biomarker Discovery, Binding Assay, SPR Assay

Journal: Phytomedicine

Article Title: Qingke Pingchuan granules alleviate airway inflammation in COPD exacerbation by inhibiting neutrophil extracellular traps in mice

doi: 10.1016/j.phymed.2024.156283

Figure Lengend Snippet: Fig. 8. QKPC and its components, EGCG and quercetin, reduce NOX2-p47phox/ROS signaling in vivo and in vitro. (A) Protein levels of NOX2 and p47phox in human neutrophils were assessed by Western blotting analysis, with relative density quantification shown as the ratio of NOX2 or p47phox to GAPDH (n = 3). (B) Protein levels of NOX2 and p47phox in lung tissues of mice were determined by Western blotting analysis and presented as the ratio of NOX2 or p47phox to GAPDH (n = 3). (C) Cellular ROS levels were measured using the oxidant-sensing probe DCFH-DA, and fluorescence intensity was recorded for each group. EGCG, quercetin, and the positive drugs NAC (20 μM, a ROS scavenger), DPI (50 μM, a general NADPH oxidase inhibitor), and GSK (50 μM, a NOX2-specific inhibitor) effectively suppressed ROS levels elevated by the combined CSE and LPS exposure (n = 6–8). (D) MDA levels in lung tissues were measured with an MDA assay kit. Data are presented as mean ± SD, with statistical significance set at p < 0.05. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. Abbreviations: CS, cigarette smoke; CSE, cigarette smoke extract; DPI, Diphenyleneiodonium chloride; EGCG, epigallocatechin-3-gallate; GSK, GSK2795039; LPS, lipopolysaccharide; MDA, malondialdehyde; NOX2, NADPH oxidase 2; p47phox (NCF1), 47 kDa neutrophil oxidase factor; QKPC, Qingke Pingchuan; Que, quercetin; ROS, reactive oxygen species.

Article Snippet: Recombinant human NOX2 protein was acquired from CUSABIO (Wuhan, China), while recombinant human P47phox protein was sourced from TargetMol Co., Ltd. (USA).

Techniques: In Vivo, In Vitro, Western Blot, Fluorescence, Multiple Displacement Amplification

Journal: Phytomedicine

Article Title: Qingke Pingchuan granules alleviate airway inflammation in COPD exacerbation by inhibiting neutrophil extracellular traps in mice

doi: 10.1016/j.phymed.2024.156283

Figure Lengend Snippet: Fig. 9. Proposed mechanisms of QKPC action against ECOPD. Abbreviations: COPD, chronic obstructive pulmonary disease; EGCG, epigallocatechin-3-gallate; LPS, lipopolysaccharide; MPO, myeloperoxidase; NE, neutrophil Elastase; NET, neutrophil extracellular trap; NOX2, NADPH oxidase 2; p47phox (NCF1), 47 kDa neutrophil oxidase factor; QKPC, Qingke Pingchuan; ROS, reactive oxygen species.

Article Snippet: Recombinant human NOX2 protein was acquired from CUSABIO (Wuhan, China), while recombinant human P47phox protein was sourced from TargetMol Co., Ltd. (USA).

Techniques:

Journal: PLoS Pathogens

Article Title: Global Functional Analyses of Cellular Responses to Pore-Forming Toxins

doi: 10.1371/journal.ppat.1001314

Figure Lengend Snippet: ( A ) Western blot of mammalian c-JUN in HaCaT cells treated with 125 ng/mL SLO for time indicated. Abbreviations: S: constitutively active version of SLO, N: non-lytic mutant N402 SLO; Co: no-SLO control. ( B ) HaCaT cells were treated with SLO and biotinylated AP-1 decoy-oligonucleotide (right panel) or biotinylated control oligonucleotide (left panel). Oligonucleotide is stained red (SA-Alexa594), and DNA is stained blue (4',6-diamidino-2-phenylindole/DAPI). The images presented are a single representative from three independent assays. Cells treated with AP-1 decoy show a greater proportion of intoxicated cells with condensed chromosomes, characteristic of becoming pyknotic ( e.g. , white arrowhead). The proportion of pyknotic cells was quantified as described in the section; the bar graph shows mean values from three independent experiments (m = mismatched oligonucleotide; AP-1 = AP-1 decoy; error bars: standard error of the mean, P value was determined with paired, one-tailed student's t-test). ( C ) A model for interconnected regulation of defense to PFT. The KGB-1 JNK-like MAPK regulates both p38 MAPK–dependent (e.g., ttm-1 , ttm-2 , UPR) and p38 MAPK-independent ( e.g. , jun-1 , kin-18 ) PFT-induced protection genes. There are also PFT protection genes that, to date, have not been linked to either MAPK pathway.

Article Snippet: For the AP-1 decoy experiment, HaCaT cells were treated in Ca 2+- free medium with 500 ng/mL SLO and 3 μM biotinylated AP-1 decoy oligodeoxynucleotide (ODN), or mismatched AP-1 decoy ODN for 30 min, (Genedetect, Auckland, New Zealand).

Techniques: Western Blot, Mutagenesis, Control, Staining, One-tailed Test

Journal: Nature

Article Title: The deubiquitinase USP9X suppresses pancreatic ductal adenocarcinoma

doi: 10.1038/nature11114

Figure Lengend Snippet: Top 20 candidate genes that cooperate with Kras G12D to promote mPDA in KCTSB13 mice CISs were scored by tumor frequency with the narrowest 15K kernel spatial distribution of insertion sites. Chr: chromosome; N: number of tumors from which the CIS was found; I, total number of insertions of the CIS in the indicated tumors.

Article Snippet: Human USP9x expression was analyzed by quantitative PCR (q-PCR) using TaqMan gene expressiom assays Hs00245009_m1 (Applied Biosystems) on a 7900HT Real-Time PCR system (Applied Biosystems).

Techniques: Mutagenesis