Journal: Poultry Science

Article Title: Development of a recombinant chimeric Newcastle disease virus-vectored vaccine conferring single-dose, triple protection against genotype VII NDV, IBDV, and H9N2 AIV

doi: 10.1016/j.psj.2026.106993

Figure Lengend Snippet: Generation and characterization of the recombinant rLaS-VIIF/HN-HA-VP2. (A) Schematic representation of the construction strategy for rLaS-VIIF/HN-HA-VP2. The open reading frames (ORFs) of the H9N2 AIV HA gene and the very virulent IBDV VP2 gene, each flanked by the non-coding region (NCR) sequences of the NDV HN gene, were inserted between the P and M genes and the F and HN genes of the pNDFL-VII-F/HN plasmid, respectively, to generate the pNDFL-VII-F/HN-HA-VP2 construct. (B) Western blot analysis of rLaS-VIIF/HN-HA-VP2 expression. Allantoic fluid harvested from embryonated eggs infected with rLaS-VIIF/HN-HA-VP2 was subjected to immunoblotting using specific antisera. Three specific bands corresponding to the uncleaved HA precursor (HA0, ∼62 kDa) and the cleaved subunits HA1 (∼47 kDa) and HA2 (∼25 kDa) were detected. Additionally, a specific band corresponding to the VP2 protein (∼42 kDa) was detected in the rLaS-VIIF/HN-HA-VP2 samples.

Article Snippet: Antibodies against IBDV were measured using the ProFLOK PLUS Infectious Bursal Disease Virus (IBD) Antibody Test Kit (Zoetis, USA) according to the manufacturer's instructions.

Techniques: Recombinant, Plasmid Preparation, Construct, Western Blot, Expressing, Infection

Journal: Poultry Science

Article Title: Development of a recombinant chimeric Newcastle disease virus-vectored vaccine conferring single-dose, triple protection against genotype VII NDV, IBDV, and H9N2 AIV

doi: 10.1016/j.psj.2026.106993

Figure Lengend Snippet: Humoral immune responses in SPF chickens immunized with rLaS-VIIF/HN-HA-VP2 and rLaS-VIIF/HN vaccines. Chickens were immunized via the ocular and nasal routes (100 µL per route) with a total of 200 µL containing 10⁶ EID₅₀ of rLaS-VIIF/HN-HA-VP2 or parental virus rLaS-VIIF/HN. Sera samples were collected before immunization and at day 10 and 20 post-immunization. (A) Antibodies against IBDV were measured using a commercial ELISA kit. (B, C) Antibodies against genotype VII NDV and H9N2 AIV were measured by hemagglutination inhibition (HI) test, respectively. Data are presented as mean ± SD; significance is denoted as * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Antibodies against IBDV were measured using the ProFLOK PLUS Infectious Bursal Disease Virus (IBD) Antibody Test Kit (Zoetis, USA) according to the manufacturer's instructions.

Techniques: Vaccines, Virus, Enzyme-linked Immunosorbent Assay, HI Assay

Journal: Poultry Science

Article Title: Development of a recombinant chimeric Newcastle disease virus-vectored vaccine conferring single-dose, triple protection against genotype VII NDV, IBDV, and H9N2 AIV

doi: 10.1016/j.psj.2026.106993

Figure Lengend Snippet: Protective efficacy of the rLaS‑VIIF/HN‑HA-VP2 vaccine against challenge with a very virulent IBDV strain. (A) Survival rates of chickens challenged with IBDV and monitored daily for 7 days post-challenge (dpc; n = 10 per group). (B) Representative gross lesions in the bursa of Fabricius at necropsy (7 dpc). Severe atrophy, accompanied by internal hemorrhage and necrosis, was evident in the bursae of birds in the rLaS‑VIIF/HN control group. In contrast, no notable gross pathological changes were observed in the rLaS‑VIIF/HN‑HA-VP2 immunized group or the uninfected controls. (C) Bursa-to-body weight index (BBIX) in immunized chickens following IBDV challenge. The mean BBIX value was significantly higher in the rLaS‑VIIF/HN‑HA-VP2 immunized group compared to the rLaS‑VIIF/HN control group (****, P < 0.0001). (D) Representative histopathological lesions in the bursa of Fabricius at 7 dpc (H&E stain; original magnification, 400 ×). Bursal tissue from birds in the rLaS‑VIIF/HN control group exhibited severe lymphoid follicle atrophy and lymphoid cell degeneration. No significant histopathological changes were observed in the rLaS‑VIIF/HN‑HA-VP2 immunized group or the uninfected controls.

Article Snippet: Antibodies against IBDV were measured using the ProFLOK PLUS Infectious Bursal Disease Virus (IBD) Antibody Test Kit (Zoetis, USA) according to the manufacturer's instructions.

Techniques: Control, Staining

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Pathology and pathogenesis of bluetongue virus serotype 24 during experimental infection in native sheep

doi: 10.3389/fcimb.2026.1710415

Figure Lengend Snippet: Detection of BTV antibodies by c-ELISA in BTV-24-infected (red line) and uninfected control (green line) animals at different days post-inoculation (DPI). Serum samples with percent inhibition (PI) values ≥ 45% were considered positive. Data are presented as mean ± SE. Asterisk (*) indicates significant difference ( p < 0.05) between groups.

Article Snippet: Twelve indigenous nondescript adult sheep of either sex aged between 2 and 3 years of approximately 30–40 kg were procured from livestock farmers after screening and negative for BTV antibodies using a competitive-enzyme linked immunosorbent assay (c-ELISA) kit (Bluetongue virus antibody cELISA test kit, VMRD Inc., Pullman, WA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Control, Inhibition

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: 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 attenuates methionine-choline-deficient diet-induced hepatic steatosis. A: Body weight of mice at the beginning and end of the experiment; B: Liver weight of mice at the beginning and end of the experiment; C: Representative hematoxylin and eosin staining images showing macrovesicular steatosis (yellow arrows), and focal inflammatory infiltration (blue arrows); scale bar: 100 μm and 20 μm; D: Representative Oil Red O staining images showing lipid droplets (red area /yellow arrows) and nuclei (blue area); scale bar: 100 μm and 20 μm; E: Quantitative analysis of Oil Red O-positive area (%); F: Serum total cholesterol levels; G: Serum triglyceride levels. Data are presented as means ± SD; a P < 0.05, b P < 0.01, c P < 0.001. MASLD: Metabolic associated steatotic liver disease; QSHXO: Qushi Huoxue ointment; 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, Staining

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: 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 attenuates methionine-choline-deficient diet-induced hepatic steatosis. A: Body weight of mice at the beginning and end of the experiment; B: Liver weight of mice at the beginning and end of the experiment; C: Representative hematoxylin and eosin staining images showing macrovesicular steatosis (yellow arrows), and focal inflammatory infiltration (blue arrows); scale bar: 100 μm and 20 μm; D: Representative Oil Red O staining images showing lipid droplets (red area /yellow arrows) and nuclei (blue area); scale bar: 100 μm and 20 μm; E: Quantitative analysis of Oil Red O-positive area (%); F: Serum total cholesterol levels; G: Serum triglyceride levels. Data are presented as means ± SD; a P < 0.05, b P < 0.01, c P < 0.001. MASLD: Metabolic associated steatotic liver disease; QSHXO: Qushi Huoxue ointment; 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, Staining

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: 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