Ketosis is an energy metabolism disorder occurring frequently in periparturient dairy cows,primarily attributed to elevated non-esterified fatty acid(NEFA)levels resulting from nega-tive energy balance(NEB).Excessive NEFA will be incompletely oxidated into large amounts of ketone bodies or be re-esterified and deposit in the liver as a consequence of hepatic limited oxida-tive capacity,ultimately leading to ketosis and fatty liver.Hypoxic microenvironments are com-monly found during the progression of various liver diseases.Hypoxia inducible factor-2 alpha(HIF-2 alpha)has been identified as a crucial regulator of lipid metabolism.However,it is still un-clear the association between HIF-2α and disrupted lipid metabolism in the livers of in ketotic cows.This study aims to investigate the effect of high concentrations of NEFA on HIF-2α expres-sion and cellular oxygen homeostasis through bovine liver tissue and primary hepatocytes.In vivo,hepatic triglyceride(TAG)content was assessed to determine the extent of hepatic lipid accumula-tion,and HIF-2α protein and mRNA levels were analyzed by immunohistochemistry staining,Western blot and qRT-PCR assay in liver tissue samples from dairy cows;in vitro,bovine primary hepatocytes were treated with different concentrations of NEFA.Oil Red O staining and TAG con-tent assay were performed to determine hepatocellular steatosis extent,and immunofluorescence staining.Western blot,and qRT-PCR were performed to analyze HIF-2α expression,in addition,lu-minescent oxygen sensor[Ru(dpp)3]Cl2 was added to indicate intracellular oxygen levels.These results showed a significant increase in TAG content and elevated HIF-2α expression in the liver tissue of ketotic cows,and high concentrations of NEFA induced lipid accumulation,upregulation of HIF-2α expression,and intracellular hypoxia in bovine primary hepatocytes.These findings sug-gested that HIF-2α was significantly"activated"in the liver of ketotic cows and high concentration of NEFA-induced bovine primary hepatocytes,and that high concentrations of NEFA induced in-tracellular hypoxia in vitro.This study provides a potential molecular target for further investiga-tion of the mechanism underlying hepatic lipid metabolism disorders in ketotic cows.

Objective To develop an animal model that replicates the clinical phenotype of severe asthma.Methods Ovalbumin(OVA)combined with IL-33 or varying doses of lipopolysaccharides(LPS)was used to explore the construction of a severe asthma mouse model.Established model animals were assessed for lung function,number of inflammatory cells,and lung tissue pathology were assessed.Expression of key genes associated with severe asthma identified from the GEO database were validated in the new model.Results Compared with OVA alone,OVA combined with IL-33 or 5 μg LPS significantly increased airway resistance and the number of inflammatory cells in bronchoalveolar lavage fluid,and aggravated the pathological damage to lung tissues.The expression patterns of key genes in the newly constructed severe asthma models were consistent with those observed in clinical patients with severe asthma.Conclusions The modeling method of combining OVA with IL-33 or LPS(5 μg)can be used to construct experimentalanimal models of severe asthma.

Pressurized intraperitoneal aerosol chemotherapy (PIPAC) is an emerging therapeutic modality for peritoneal carcinomatosis. This review aims to evaluate the safety, efficacy, and current clinical application of PIPAC in the treatment of peritoneal metastases originating from gastrointestinal malignancies. This review outlines the technical principles, historical development, procedural steps, commonly used drugs, and administration protocols of PIPAC; analyses the current clinical application status of PIPAC technology; discusses the current challenges and future directions of PIPAC; suggests that PIPAC technology still needs to conduct more high-quality and large-sample clinical trials to further establish the safety and efficacy of PIPAC, optimize its indications and formulate standardized operation specifications. In the future, multi-centre cooperation, multi-disciplinary cooperation, precision medicine strategies and new drug research and development will promote the clinical transformation and standardized application of PIPAC technology.

Objective:This study aimed to summarize and analyze the preliminary application experience of a novel pressurized intraperitoneal aerosol chemotherapy (PIPAC) device in patients with peritoneal metastases of gastrointestinal malignancies.Methods:In this descriptive case series study, four patients with pathologically confirmed peritoneal metastatic gastrointestinal malignancies were enrolled, receiving PIPAC treatment at Guangdong Provincial People's Hospital from December 2024 to February 2025. The PIPAC treatment was performed five times on these patients . Key procedural steps included equipment preparation, operation platform construction, cytologic examination of ascites, Peritoneal Cancer Index (PCI) scoring, biopsy of peritoneal metastases, drug preparation, connection and testing of the PIPAC device, PIPAC treatment, and aerosol evacuation. Preoperative and postoperative clinical data were collected and analyzed to assess the safety and feasibility of PIPAC treatment.Results:3 males and 1 female patients were enrolled; median age was 57 (range, 36-70) years old; median body mass index was 22.5 (range, 18.0-24.6) kg/m2, and the preoperative Eastern Cooperative Oncology Group (ECOG) score was 1 for all cases. Four patients successfully completed the five PIPAC treatments without any intraoperative adverse events. The PIPAC treatment time ranged from 34 minutes to 36 minutes, with a median preoperative PCI score of 18 (range, 5-25). The average Peritoneal Regression Grading Score (PRGS) before the first and second PIPAC treatments were 2.1±0.8 and 1.7±0.6, respectively. The median Visual Analog Scale (VAS) scores for pain on postoperative days 1, 2, and 3 were 2 (range, 2-4), 1 (range, 0-2), and 1 (range, 0-2), respectively. All patients resumed the oral intake and ambulation on the first postoperative day, with a postoperative hospital stay of 3 days. No postoperative complications or perioperative death occurred.Conclusion:The preliminary study results indicate that the use of the novel PIPAC device for the treatment of peritoneal metastases of gastrointestinal malignant tumors is safe and feasible.

Objective To establish a mouse model of H1N1 influenza wind-heat syndrome by combining climate intervention with influenza virus nasal drops.Methods Seventy-two BALB/c mice were divided randomly into nine groups:a Control group,wind-heat(FR)groups(FR-3Day,FR-5Day),and Model groups(1LD-3Day,2LD-3Day,3LD-3Day,1LD-5Day,2LD-5Day,2LD-5Day,3LD-5Day)(n=8 mice per group).Mice in the Control group were housed in a normal environment,while mice in the FR and Model groups were kept in wind-heat conditions for 7 d.Mice in the Model groups received nasal PR8 influenza virus infection on the 8th day,and mice in the Control and FR heat groups received equal amounts of physiological saline nasal drops.After virus challenge,each group was housed in a normal environment and samples were taken on days 3 and 5.The appearance of the mice was observed and recorded and the lung index,routine blood parameters,lung tissue pathology,serum interleukin(IL)-6 levels,and virus titers were detected in each group based on their behavioral status,stools,and body temperature.Results After 7 d of wind-heat intervention,mice in the FR groups showed no significant abnormalities in terms of appearance,stools,body temperature,routine blood parameters,or lung tissue pathology compared with the Control group.The appearance,lung index,red blood cell count,hemoglobin,hematocrit,pathological result,and body temperature in the Model groups worsened progressively with increasing time and toxin dosage,while the neutrophil percentage,lymphocyte percentage,virus titer,and serum IL-6 levels peaked on day 3 after viral attack,for the same viral dose,and then decreased slightly on day 5.Conclusions PR8 nasal drops and 7 d of wind-heat climate intervention can be used to establish a mouse model of influenza wind-heat syndrome.

Objective To observe the effect of pirfenidone on myocardial fibrosis in rats and inves-tigate its underlying mechanism.Methods Twenty-four SD rats were randomly divided into sham-operation group,model group,low-and high-dose pirfenidone groups,with 6 rats in each group.Rat model of myocardial fibrosis was established by injecting isoprenaline into the tail vein,while normal saline was given to the sham operation group.Pirfenidone of 150 and 300 mg/(kg·d)were infused gastrically to the rats of low-and high-dose pirfenidone groups after modeling.Mas-son staining was used to observe the severity of myocardial fibrosis,immunohistochemical assay was employed to detect the expression of Collagen-1,NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome,cysteinyl aspartate-specific protease-1(Caspase-1),and Western blotting was performed to detect the protein levels of Collogen-1 and atrial desmo-plakin D(gasdermin D,GSDMD).Results The model group showed obvious myocardial fibrosis,and elevated expression of Collogen-1,NLRP3,Caspase-1 and GSDMD when compared with the sham operation group(P＜0.05).Low-and high-dose pirfenidone treatment resulted in signifi-cantly reduced myocardial fibrosis and reduced expression of Collogen-1,NLRP3,Caspase-1 and GSDMD[(8.14±1.40)％,(6.56±0.75)％vs(22.15±2.57)％,P＜0.05;0.14±0.03 vs 0.33±0.05,0.42±0.13,P＜0.05;(10.34±1.40)％,(10.33±3.40)％vs(23.22±1.99)％,P＜0.05;(15.67±0.56)％,(17.33±0.78)％vs(22.87±1.92)％,P＜0.05;0.43±0.06,0.46±0.11 vs 0.65±0.03,P＜0.05].Conclusion Pirfenidone inhibits cardiomyocyte pyroptosis and attenuates myocar-dial fibrosis through the NLRP3/Caspase-1/GSDMD signaling axis.

Ketosis is an energy metabolism disorder occurring frequently in periparturient dairy cows,primarily attributed to elevated non-esterified fatty acid(NEFA)levels resulting from nega-tive energy balance(NEB).Excessive NEFA will be incompletely oxidated into large amounts of ketone bodies or be re-esterified and deposit in the liver as a consequence of hepatic limited oxida-tive capacity,ultimately leading to ketosis and fatty liver.Hypoxic microenvironments are com-monly found during the progression of various liver diseases.Hypoxia inducible factor-2 alpha(HIF-2 alpha)has been identified as a crucial regulator of lipid metabolism.However,it is still un-clear the association between HIF-2α and disrupted lipid metabolism in the livers of in ketotic cows.This study aims to investigate the effect of high concentrations of NEFA on HIF-2α expres-sion and cellular oxygen homeostasis through bovine liver tissue and primary hepatocytes.In vivo,hepatic triglyceride(TAG)content was assessed to determine the extent of hepatic lipid accumula-tion,and HIF-2α protein and mRNA levels were analyzed by immunohistochemistry staining,Western blot and qRT-PCR assay in liver tissue samples from dairy cows;in vitro,bovine primary hepatocytes were treated with different concentrations of NEFA.Oil Red O staining and TAG con-tent assay were performed to determine hepatocellular steatosis extent,and immunofluorescence staining.Western blot,and qRT-PCR were performed to analyze HIF-2α expression,in addition,lu-minescent oxygen sensor[Ru(dpp)3]Cl2 was added to indicate intracellular oxygen levels.These results showed a significant increase in TAG content and elevated HIF-2α expression in the liver tissue of ketotic cows,and high concentrations of NEFA induced lipid accumulation,upregulation of HIF-2α expression,and intracellular hypoxia in bovine primary hepatocytes.These findings sug-gested that HIF-2α was significantly"activated"in the liver of ketotic cows and high concentration of NEFA-induced bovine primary hepatocytes,and that high concentrations of NEFA induced in-tracellular hypoxia in vitro.This study provides a potential molecular target for further investiga-tion of the mechanism underlying hepatic lipid metabolism disorders in ketotic cows.

Objective To develop an animal model that replicates the clinical phenotype of severe asthma.Methods Ovalbumin(OVA)combined with IL-33 or varying doses of lipopolysaccharides(LPS)was used to explore the construction of a severe asthma mouse model.Established model animals were assessed for lung function,number of inflammatory cells,and lung tissue pathology were assessed.Expression of key genes associated with severe asthma identified from the GEO database were validated in the new model.Results Compared with OVA alone,OVA combined with IL-33 or 5 μg LPS significantly increased airway resistance and the number of inflammatory cells in bronchoalveolar lavage fluid,and aggravated the pathological damage to lung tissues.The expression patterns of key genes in the newly constructed severe asthma models were consistent with those observed in clinical patients with severe asthma.Conclusions The modeling method of combining OVA with IL-33 or LPS(5 μg)can be used to construct experimentalanimal models of severe asthma.

Pressurized intraperitoneal aerosol chemotherapy (PIPAC) is an emerging therapeutic modality for peritoneal carcinomatosis. This review aims to evaluate the safety, efficacy, and current clinical application of PIPAC in the treatment of peritoneal metastases originating from gastrointestinal malignancies. This review outlines the technical principles, historical development, procedural steps, commonly used drugs, and administration protocols of PIPAC; analyses the current clinical application status of PIPAC technology; discusses the current challenges and future directions of PIPAC; suggests that PIPAC technology still needs to conduct more high-quality and large-sample clinical trials to further establish the safety and efficacy of PIPAC, optimize its indications and formulate standardized operation specifications. In the future, multi-centre cooperation, multi-disciplinary cooperation, precision medicine strategies and new drug research and development will promote the clinical transformation and standardized application of PIPAC technology.

Objective:This study aimed to summarize and analyze the preliminary application experience of a novel pressurized intraperitoneal aerosol chemotherapy (PIPAC) device in patients with peritoneal metastases of gastrointestinal malignancies.Methods:In this descriptive case series study, four patients with pathologically confirmed peritoneal metastatic gastrointestinal malignancies were enrolled, receiving PIPAC treatment at Guangdong Provincial People's Hospital from December 2024 to February 2025. The PIPAC treatment was performed five times on these patients . Key procedural steps included equipment preparation, operation platform construction, cytologic examination of ascites, Peritoneal Cancer Index (PCI) scoring, biopsy of peritoneal metastases, drug preparation, connection and testing of the PIPAC device, PIPAC treatment, and aerosol evacuation. Preoperative and postoperative clinical data were collected and analyzed to assess the safety and feasibility of PIPAC treatment.Results:3 males and 1 female patients were enrolled; median age was 57 (range, 36-70) years old; median body mass index was 22.5 (range, 18.0-24.6) kg/m2, and the preoperative Eastern Cooperative Oncology Group (ECOG) score was 1 for all cases. Four patients successfully completed the five PIPAC treatments without any intraoperative adverse events. The PIPAC treatment time ranged from 34 minutes to 36 minutes, with a median preoperative PCI score of 18 (range, 5-25). The average Peritoneal Regression Grading Score (PRGS) before the first and second PIPAC treatments were 2.1±0.8 and 1.7±0.6, respectively. The median Visual Analog Scale (VAS) scores for pain on postoperative days 1, 2, and 3 were 2 (range, 2-4), 1 (range, 0-2), and 1 (range, 0-2), respectively. All patients resumed the oral intake and ambulation on the first postoperative day, with a postoperative hospital stay of 3 days. No postoperative complications or perioperative death occurred.Conclusion:The preliminary study results indicate that the use of the novel PIPAC device for the treatment of peritoneal metastases of gastrointestinal malignant tumors is safe and feasible.