Journal: World Journal of Hepatology

Article Title: Qushi Huoxue ointment ameliorates metabolic associated steatotic liver disease through autophagy activation and ferroptosis inhibition

doi: 10.4254/wjh.v18.i2.115763

Figure Lengend Snippet: Experimental design summary diagram. Metabolic associated steatotic liver disease mouse model was first established using the methionine-choline-deficient diet. Following Qushi Huoxue ointment (QSHXO) treatment, serum and liver tissue samples were collected to assess pathological changes, liver function markers, and inflammatory cytokine levels. Subsequently, the bioactive components of QSHXO in serum were identified using liquid chromatography-mass spectrometry/mass spectrometry analysis. Next, network pharmacology was applied to predict the potential target pathways of QSHXO in the treatment of metabolic associated steatotic liver disease, specifically focusing on autophagy and ferroptosis. These predicted targets were further validated through western blot, quantitative reverse-transcription polymerase chain reaction, immunohistochemistry, and transmission electron microscopy. MCD: Methionine-choline-deficient; MASLD: Metabolic associated steatotic liver disease; QSHXO: Qushi Huoxue ointment; TNF-α: Tumor necrosis factor-α; IL-β: Interleukin-β; ELISA: Enzyme-linked immunosorbent assay; ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; TC: Total cholesterol; TG: Triglyceride; Nrf2: Nuclear factor erythroid 2-related factor 2; Lc3: Light chain 3; GPX4: Glutathione peroxidase 4.

Article Snippet: The following reagents and antibodies were used in this study: Alanine aminotransferase (ALT) test kit (C009-2-1, Nanjing Jiancheng, Nanjing, Jiangsu Province, China); aspartate aminotransferase (AST) test kit (C010-2-1, Nanjing Jiancheng, Nanjing, Jiangsu Province, China); total cholesterol (TC) test kit (A111-1-1, Nanjing Jiancheng, Nanjing, Jiangsu Province, China); triglyceride (TG) test kit (A110-1-1, Nanjing Jiancheng, Nanjing, Jiangsu Province, China); tumor necrosis factor-α (TNF-α, EK201BHS, LianKe Bio, Hangzhou, Zhejiang Province, China); interleukin (IL)-1β (EK282/4, LianKe Bio, Hangzhou, Zhejiang Province, China); oltipraz (B5958, APExBIO, TX, United States); Beclin1 (AF5128, 1:1000, Affinity, Shanghai, China); light chain 3 (LC3) (AF5402, 1:1000, Affinity, Shanghai, China); P62 (AF5384, 1:1000, Affinity, Shanghai, China); Nrf2 (AF0639, 1:800, Affinity, Shanghai, China); glutathione peroxidase 4 (GPX4) (A11243, 1:1000, Abclonal, Wuhan, Hubei Province, China); SLC7A11 (A2413, 1:1000, Abclonal, Wuhan, Hubei Province, China); and lamin B1 (BF8009, 1:2000, Affinity, Shanghai, China).

Techniques: Ointment, Liquid Chromatography, Mass Spectrometry, Western Blot, Reverse Transcription, Polymerase Chain Reaction, Immunohistochemistry, Transmission Assay, Electron Microscopy, Enzyme-linked Immunosorbent Assay

Journal: World Journal of Hepatology

Article Title: Qushi Huoxue ointment ameliorates metabolic associated steatotic liver disease through autophagy activation and ferroptosis inhibition

doi: 10.4254/wjh.v18.i2.115763

Figure Lengend Snippet: Qushi Huoxue ointment ameliorates liver injury and inflammatory response in metabolic associated steatotic liver disease mice. A: Serum levels of aspartate aminotransferase, alanine aminotransferase, tumor necrosis factor-α, and interleukin-β; B: Schematic diagram illustrating the relationship between hepatic lipid accumulation and liver injury. Data are presented as means ± SD; a P < 0.05, c P < 0.001. MASLD: Metabolic associated steatotic liver disease; QSHXO: Qushi Huoxue ointment; AST: Aspartate aminotransferase; ALT: Alanine aminotransferase; TNF-α: Tumor necrosis factor-α; IL-β: Interleukin-β; TC: Total cholesterol; TG: Triglyceride.

Article Snippet: The following reagents and antibodies were used in this study: Alanine aminotransferase (ALT) test kit (C009-2-1, Nanjing Jiancheng, Nanjing, Jiangsu Province, China); aspartate aminotransferase (AST) test kit (C010-2-1, Nanjing Jiancheng, Nanjing, Jiangsu Province, China); total cholesterol (TC) test kit (A111-1-1, Nanjing Jiancheng, Nanjing, Jiangsu Province, China); triglyceride (TG) test kit (A110-1-1, Nanjing Jiancheng, Nanjing, Jiangsu Province, China); tumor necrosis factor-α (TNF-α, EK201BHS, LianKe Bio, Hangzhou, Zhejiang Province, China); interleukin (IL)-1β (EK282/4, LianKe Bio, Hangzhou, Zhejiang Province, China); oltipraz (B5958, APExBIO, TX, United States); Beclin1 (AF5128, 1:1000, Affinity, Shanghai, China); light chain 3 (LC3) (AF5402, 1:1000, Affinity, Shanghai, China); P62 (AF5384, 1:1000, Affinity, Shanghai, China); Nrf2 (AF0639, 1:800, Affinity, Shanghai, China); glutathione peroxidase 4 (GPX4) (A11243, 1:1000, Abclonal, Wuhan, Hubei Province, China); SLC7A11 (A2413, 1:1000, Abclonal, Wuhan, Hubei Province, China); and lamin B1 (BF8009, 1:2000, Affinity, Shanghai, China).

Techniques: Ointment

Journal: Cell Genomics

Article Title: Spatial transcriptomics identifies novel Pseudomonas aeruginosa virulence factors

doi: 10.1016/j.xgen.2025.100805

Figure Lengend Snippet:

Article Snippet: cKit , Novus Biologicals , Clone: 9D2 NB110-93600AF532 PRID: AB_3181352.

Techniques: Virus, Recombinant, Software

Journal: Cell death & disease

Article Title: CHCHD2 rescues the mitochondrial dysfunction in iPSC-derived neurons from patient with Mohr-Tranebjaerg syndrome.

doi: 10.1038/s41419-025-07472-9

Figure Lengend Snippet: Fig. 3 Dysfunction of mitochondria in iPSC-derived neurons due to mutation in TIMM8A. A, B No differences in HSP60, TOMM40, and TIMM23 levels were observed among CTRL-, MUT-, and MTS-neurons by Western blot. Relative protein levels were quantified and normalized to β-Tubulin as a loading control. n = 3 independent biological replicates. C, D. Western blot analysis showed decreased levels of COX4 in mitochondrial complex IV in MUT- and MTS-neurons. Relative levels of COX4 were quantified and normalized to HSP60 as a loading control. n = 3 independent biological replicates. E The activity of mitochondrial complex IV was decreased in MUT- and MTS-neurons compared to CTRL-neurons. n = 3 independent biological replicates. F ATP production was decreased in MUT- and MTS-neurons compared to CTRL- neurons. n = 3 independent biological replicates. G ROS levels were increased in MUT- and MTS-neurons compared to CTRL-neurons. n = 3 independent biological replicates. H, I Levels of Cyt c and cleaved caspase-3 in CTRL-, MUT-, and MTS-neurons after treatment with 100 µM TBHP for 24 h were analyzed by Western blot. Relative protein levels of Cyt c and cleaved caspase-3 in the cytosol were quantified and normalized to β-Tubulin as a loading control. n = 3 independent biological replicates. Data are represented as mean ± SD. Significance was determined by one-way ANOVA with Tukey’s post hoc test: ns, not significant; *p < 0.05; **p < 0.01; ****p < 0.0001 (applied to panels B, D–G, and I).

Article Snippet: Activity analysis of mitochondrial complex IV The activity of mitochondrial complex IV was measured using the Cell Mitochondrial Complex IV (Cytochrome C Oxidase) Activity Assay Kit (Elabscience, Cat# E-BC-K837-M) according to the manufacturer’s protocol.

Techniques: Derivative Assay, Mutagenesis, Western Blot, Control, Activity Assay

Journal: Cell death & disease

Article Title: CHCHD2 rescues the mitochondrial dysfunction in iPSC-derived neurons from patient with Mohr-Tranebjaerg syndrome.

doi: 10.1038/s41419-025-07472-9

Figure Lengend Snippet: Fig. 4 Altered gene expression in iPSC-derived neurons revealed by RNA sequencing. A A total of 2,843 genes were significantly upregulated, and 1,667 genes were significantly downregulated in MTS-neurons compared to CTRL-neurons (adjusted p < 0.05 and |log2(fold change)| > 1). Top altered genes, including CHCHD2, are labeled. B Top 10 altered biological processes in MTS-neurons compared to CTRL-neurons, identified by Gene Ontology (GO) enrichment analysis (adjusted p < 0.05). C Top 10 altered Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways in MTS-neurons compared to CTRL-neurons (adjusted p < 0.05). D Representative differentially expressed genes involved in mitochondria-related apoptosis. E Western blot analysis of the anti-apoptotic protein BCL-2 and the pro-apoptotic protein BAX in isolated mitochondrial lysates from CTRL- and MTS-neurons. F Western blot analysis of CHCHD2 in isolated mitochondrial lysates from CTRL-, MUT-, and MTS-neurons. G Expression levels of BCL-2 and BAX were quantified and normalized to HSP60 as a loading control. n = 3 independent biological replicates. Data are represented as mean ± SD. Significance was determined by two-tailed unpaired Student’s t test: ns, not significant; **p < 0.01. H CHCHD2 expression levels were quantified and normalized to HSP60 as a loading control. n = 3 independent biological replicates. Data are represented as mean ± SD. Significance was determined by one-way ANOVA with Tukey’s post hoc test: **p < 0.01; ****p < 0.0001.

Article Snippet: Activity analysis of mitochondrial complex IV The activity of mitochondrial complex IV was measured using the Cell Mitochondrial Complex IV (Cytochrome C Oxidase) Activity Assay Kit (Elabscience, Cat# E-BC-K837-M) according to the manufacturer’s protocol.

Techniques: Gene Expression, Derivative Assay, RNA Sequencing, Labeling, Western Blot, Isolation, Expressing, Control, Two Tailed Test

Journal: Cell death & disease

Article Title: CHCHD2 rescues the mitochondrial dysfunction in iPSC-derived neurons from patient with Mohr-Tranebjaerg syndrome.

doi: 10.1038/s41419-025-07472-9

Figure Lengend Snippet: Fig. 6 CHCHD2 overexpression rescues mitochondrial morphology defects in iPSC-derived neurons. A, B Mitochondrial morphology in CTRL- and MTS-neurons at day 30, as revealed by transmission electron microscopy (TEM). Scale bar: 1 μm. n = 3 independent biological replicates. Quantification of mitochondrial perimeter, area, aspect ratio, and form factor was performed (103 mitochondria for CTRL group; 102 for MTS group). C, D Mitochondrial morphology in the Vector group and CHCHD2-Flag group of MTS-neurons at day 30, as revealed by TEM. Scale bar: 1 μm. n = 3 independent biological replicates. Quantification of mitochondrial perimeter, area, aspect ratio, and form factor was performed (107 mitochondria for Vector group; 124 for CHCHD2-Flag group). Data are represented as mean ± SD. Significance was determined by two-tailed unpaired Student’s t test: **p < 0.01; ****p < 0.0001.

Article Snippet: Activity analysis of mitochondrial complex IV The activity of mitochondrial complex IV was measured using the Cell Mitochondrial Complex IV (Cytochrome C Oxidase) Activity Assay Kit (Elabscience, Cat# E-BC-K837-M) according to the manufacturer’s protocol.

Techniques: Over Expression, Derivative Assay, Transmission Assay, Electron Microscopy, Plasmid Preparation, Two Tailed Test