Recent studies have indicated that the expression of ubiquitin-specific protease 51 (USP51), a novel deubiquitinating enzyme (DUB) that mediates protein degradation as part of the ubiquitin‒proteasome system (UPS), is associated with tumor progression and therapeutic resistance in multiple malignancies. However, the underlying mechanisms and signaling networks involved in USP51-mediated regulation of malignant phenotypes remain largely unknown. The present study provides evidence of USP51's functions as the prominent DUB in chemoresistant triple-negative breast cancer (TNBC) cells. At the molecular level, ectopic expression of USP51 stabilized the 78 kDa Glucose-Regulated Protein (GRP78) protein through deubiquitination, thereby increasing its expression and localization on the cell surface. Furthermore, the upregulation of cell surface GRP78 increased the activity of ATP binding cassette subfamily B member 1 (ABCB1), the main efflux pump of doxorubicin (DOX), ultimately decreasing its accumulation in TNBC cells and promoting the development of drug resistance both in vitro and in vivo. Clinically, we found significant correlations among USP51, GRP78, and ABCB1 expression in TNBC patients with chemoresistance. Elevated USP51, GRP78, and ABCB1 levels were also strongly associated with a poor patient prognosis. Importantly, we revealed an alternative intervention for specific pharmacological targeting of USP51 for TNBC cell chemosensitization. In conclusion, these findings collectively indicate that the USP51/GRP78/ABCB1 network is a key contributor to the malignant progression and chemotherapeutic resistance of TNBC cells, underscoring the pivotal role of USP51 as a novel therapeutic target for cancer management.

Objective To investigate the anti-hepatocellular carcinoma mechanism of Chloranthus fortunei(A.Gray)Solms-Laub.via network pharmacology,molecular docking techniques and in vitro experiments.Methods Chemical composition of Chloranthus fortunei(A.Gray)Solms-Laub.was searched by literature.Swiss Target Prediction was used to find corresponding targets.STRING was used to construct protein-protein interactions network(PPI).DAVID was used to enrich GO analysis and KEGG pathway.AutoDock Vina 1.1.2 and Pymol visualisation was used for docking and validation.Results Chloranthus fortunei(A.Gray)Solms-Laub.had 61 active components,685 targets,and 279 intersections with disease targets.The PPI showed that the main active components were Luteolin,Chloranthalactone C,Shizukanolide H,Esculetin,7-Hydroxycoumarin.The key targets were GAPDH,VEGFA,STAT3,JUN,HSP90AA1,AKT1,CTNNB1,CASP3,and ALB.Biological process(BP)involved protein phosphorylation,signal transduction,regulation of RNA polymerase II promoter transcription,cell proliferation,apoptosis.Cellular component(CC)involved cytoplasm,nucleus,cell membrane,cellular exosome.Molecular function(MF)involves protein binding,ATPase,threonine kinase,protein kinase activity.KEGG involved cancer pathway,metabolic pathway,PI3K-Akt signalling pathway,cancer proteoglycans,lipids and atherosclerosis,cytomegalovirus infection,microRNAs in cancer,human T-cell leukaemia virus type 1,Ras signalling pathway,MAPK signalling pathway.Molecular docking showed that silverweed lactone H had a strong affinity for each of the other target proteins,indicating that this component plays a key role.The results of RT-qPCR assay and WB assay showed that there were significant differences in gene and protein expression levels before and after drug administration.Conclusion The Chinese medicine in Chloranthus fortunei(A.Gray)Solms-Laub.can treat hepatocellular carcinoma through the MAPK pathway,and the main active ingredients have good docking effects with the core target proteins of the disease.

Objective To establish a stable,reliable,and clinically relevant porcine model of endotoxin-induced acute respiratory distress syndrome(ARDS).Methods Ten 8-month-old male Bama minipigs were deeply sedated,followed by invasive mechanical ventilation and electrocardiographic monitoring.Lipopolysaccharide(LPS)was intravenously pumped at 600 μg/(kg·h)for 3 hours,then maintained at 15 μg/(kg·h)thereafter.Dynamic monitoring was performed at five time points after LPS injection(LPS 0,1,3,5,and 8 h),including arterial blood gas analysis and chest computed tomography(CT)scans.Pathological examination of lung tissues obtained via bronchoscopic biopsy(HE staining and transmission electron microscopy)was conducted.These indicators were comprehensively used to evaluate the success of the animal model.Results At 5 hours after LPS administration,8 minipigs developed symptoms such as skin cyanosis,elevated body temperature,and respiratory distress.The oxygenation index decreased to<300 mmHg.Chest CT scans showed diffuse pulmonary infiltrates.Histopathology revealed alveolar edema and hyaline membrane formation.Transmission electron microscopy demonstrated disruption of pulmonary blood-air barrier,depletion of lamellar bodies in type Ⅱ pneumocytes,inflammatory cell infiltration,and exudation of plasma proteins and fibrin.Compared with LPS 0 h,at LPS 8 h,the oxygenation index and arterial blood pH were significantly decreased(P<0.001),while blood lactic acid and serum potassium were significantly increased(P<0.05);serum calcium and base excess were significantly decreased(P<0.05),and the lung injury score based on HE-stained lung sections was significantly increased(P<0.01).Conclusion The porcine ARDS model established by continuous LPS injection can dynamically simulate the pathophysiological characteristics and typical pathological manifestations of clinical septic ARDS,making it an effective tool to study the pathogenesis,prevention,and treatment strategies of septic ARDS.

Objective To observe the effects of Yiqi Huoxue Prescription in regulating METTL3/HAND2/TGF-β1 signaling axis on rats with qi deficiency and blood stasis syndrome of coronary heart disease.Methods Totally 48 male SD rats were randomly divided into sham-operation group,model group,Yiqi Huoxue Prescription group and Western medicine(isosorbide mononitrate)group,with 12 rats in each group.Except for the sham-operation group,the animal models of coronary heart disease with qi deficiency and blood stasis syndrome were prepared by the method of ligation of the left anterior descending branch of coronary artery combined with swimming exhaustion.After modeling,Yiqi Huoxue Prescription group and the Western medicine group were gavaged for 28 d.The general conditions of the rats were observed,and electrocardiograms and echocardiograms were collected to detect the cardiac function of the rats,blood rheology was used to detect the degree of blood stasis,HE staining was used to observe the morphology of myocardial tissue,immunohistochemistry was used to detect the expression of METTL3,TGF-β1,Smad2 and Smad3 in myocardial tissue,RT-PCR and Western blot were used to detect the expression of METTL3,HAND2,TGF-β1,Smad2 and Smad3 mRNA and protein in myocardial tissue.Results Compared with the sham-operation group,the model group rats showed symptoms of qi deficiency such as mental lethargy,slow movement,drowsiness,and decreased body mass,significant elevation of ST segment on electrocardiogram,the left ventricular ejection fraction(LVEF)and left ventricular fractional shortening(LVFS)decreased(P＜0.01),the left ventricular end diastolic volume(LVEDV),left ventricular end systolic volume(LVESV),left ventricular end diastolic diameter(LVEDd)and left ventricular end systolic diameter(LVEDs)increased(P＜0.01),the red blood cell aggregation index,Carcasson viscosity and whole blood viscosity increased(P＜0.01);with enlargement and necrosis of myocardial cells,disordered arrangement of myocardial fibers,and infiltration of inflammatory cells into myocardial tissue,the mRNA and protein expressions of METTL3,TGF-β1,Smad2 and Smad3 in myocardial tissue increased(P＜0.05,P＜0.01),while the mRNA and protein expression of HAND2 decreased(P＜0.01).Compared with the model group,the Yiqi Huoxue Prescription group and the Western medicine group showed significant improvement in the mental state of rats,significant ST segment regression on electrocardiogram,LVEF and LVFS increased(P＜0.01),LVEDV,LVESV,LVEDd and LVEDs decreased(P＜0.05,P＜0.01),with red blood cell aggregation index,Carcasson viscosity and whole blood viscosity decreased(P＜0.01);the arrangement of myocardial fibers was neat,the structure and morphology of myocardial cells were improved,the infiltration of inflammatory cells was reduced,the mRNA and protein expressions of METTL3,TGF-β1,Smad2 and Smad3 in myocardial tissue decreased(P＜0.05,P＜0.01),and the mRNA and protein expression of HAND2 increased(P＜0.05,P＜0.01).Conclusion Yiqi Huoxue Prescription can alleviate the pathological damage of myocardial tissue in rats with coronary heart disease of qi deficiency and blood stasis syndrome,improve heart function and inhibit inflammatory response.Its mechanism may be related to the regulation of the METTL3/HAND2/TGF-β1 signaling axis.

AIM To explore the differential metabolites of different aged Citrus reticulate'Chachi'and their processed cakes.METHODS Non-targeted metabolomics technology of GC-TOF-MS was used to analyze the chemical constituents.The data was processed by principal component analysis and orthogonal partial least squares discriminant analysis,and the differential metabolites were identified.RESULTS A total of 74 differential metabolites were identified,including 16 glycosides,14 organic acids and their derivatives,11 amino acids and their derivatives,and 4 flavonoids.Comparative analysis revealed 40 and 30 differential metabolites between fresh C.reticulate'℃hachi'and 3-year or 5-year aged samples,respectively.Furthermore,27 and 34 differential metabolites were identified between the 3-year or 5-year aged samples and their corresponding processed cakes,respectively.Differential metabolites among fresh,aged C.reticulate'Chachi',and processed cakes were predominantly enriched in 6 metabolic pathways,including the biosynthesis of secondary metabolites.Specifically,differential metabolites between 3-year aged C.reticulate'Chachi'and its processed cake were significantly enriched in 4 pathways,such as ABC transporters.Differential metabolites between 5-year aged C.reticulate'Chachi'and its processed cake were mainly enriched in 5 pathways,including carbon metabolism.CONCLUSION Non-targeted metabolomics technology can elucidate the chemical compositional differences among fresh/aged and processed cakes of C.reticulate'Chachi',laying a foundation for the research into C.reticulate'Chachi'aging processing techniques and the development of processed products.

Objective To understand the current situation of healthcare-associated infection(HAI)in China,pro-vide data support and decision-making basis for formulating scientific and effective strategies for HAI prevention and control.Methods A nationwide cross-sectional survey on HAI was conducted among various types and levels of medical institutions in China according to a unified protocol of bedside surveys and case investigations.Results In 2024,a total of 5 736 medical institutions and 2 751 765 patients were surveyed.Among them,34 889 HAI cases were identified,with a prevalence rate of 1.27％.The number of HAI episodes was 38 032,and case prevalence rate was 1.38％.The prevalence rate of HAI in medical institutions in different regions of China ranged from 0.66％to 2.35％.Among medical institutions of different scales,those with a bed capacity of ≥900 had the high-est incidence of HAI,reaching 1.65％.The most common infection site was the lower respiratory tract(44.66％),followed by the urinary tract(12.94％),surgical site(9.32％),upper respiratory tract(7.02％),and bloodstream infection(5.78％).The top 3 departments with the highest HAI rates were the general intensive care unit(10.02％),department of neurosurgery(5.51％),and department(group)of hematology(5.34％).A total of 23 238 strains of HAI pathogens were detected,with 10 714 strains(46.10％)from lower respiratory tract speci-mens.The top 5 detected strains were Klebsiella pneumoniae(14.76％),Pseudomonas aeruginosa(13.33％),Escherichia coli(12.79％),Acinetobacter baumannii(9.23％),and Staphylococcus aureus(7.88％).231 944 pa-tients underwent class Ⅰ incision surgery were monitored,with 1 647 cases experienced surgical site infection,and the prevalence rate of surgical site infection was 0.71％.The number of patients who should undergo pathogen de-tection(patients receiving therapeutic and therapeutic combined prophylactic antimicrobial agents)was 715 179,while the actual number was 480 492,with a pathogen detection rate of 67.18％.425 225 patients received patho-genic detection before treatment,with a detection rate of 59.46％.Conclusion The overall HAI prevalence in Chi-na is lower,showing disparities among medical institutions of different regions and scales.Therefore,precise imple-mentation of measures is necessary for HAI prevention and control,with a focus on high-risk institutions and high-risk departments,key areas,and critical procedures.All levels of medical institutions should continuously reduce the incidence of HAI by strengthening monitoring,standardizing the use of antimicrobial agents,and reinforcing basic HAI prevention and control measures.

AIM To explore the differential metabolites of different aged Citrus reticulate'Chachi'and their processed cakes.METHODS Non-targeted metabolomics technology of GC-TOF-MS was used to analyze the chemical constituents.The data was processed by principal component analysis and orthogonal partial least squares discriminant analysis,and the differential metabolites were identified.RESULTS A total of 74 differential metabolites were identified,including 16 glycosides,14 organic acids and their derivatives,11 amino acids and their derivatives,and 4 flavonoids.Comparative analysis revealed 40 and 30 differential metabolites between fresh C.reticulate'℃hachi'and 3-year or 5-year aged samples,respectively.Furthermore,27 and 34 differential metabolites were identified between the 3-year or 5-year aged samples and their corresponding processed cakes,respectively.Differential metabolites among fresh,aged C.reticulate'Chachi',and processed cakes were predominantly enriched in 6 metabolic pathways,including the biosynthesis of secondary metabolites.Specifically,differential metabolites between 3-year aged C.reticulate'Chachi'and its processed cake were significantly enriched in 4 pathways,such as ABC transporters.Differential metabolites between 5-year aged C.reticulate'Chachi'and its processed cake were mainly enriched in 5 pathways,including carbon metabolism.CONCLUSION Non-targeted metabolomics technology can elucidate the chemical compositional differences among fresh/aged and processed cakes of C.reticulate'Chachi',laying a foundation for the research into C.reticulate'Chachi'aging processing techniques and the development of processed products.

Objective To investigate the anti-hepatocellular carcinoma mechanism of Chloranthus fortunei(A.Gray)Solms-Laub.via network pharmacology,molecular docking techniques and in vitro experiments.Methods Chemical composition of Chloranthus fortunei(A.Gray)Solms-Laub.was searched by literature.Swiss Target Prediction was used to find corresponding targets.STRING was used to construct protein-protein interactions network(PPI).DAVID was used to enrich GO analysis and KEGG pathway.AutoDock Vina 1.1.2 and Pymol visualisation was used for docking and validation.Results Chloranthus fortunei(A.Gray)Solms-Laub.had 61 active components,685 targets,and 279 intersections with disease targets.The PPI showed that the main active components were Luteolin,Chloranthalactone C,Shizukanolide H,Esculetin,7-Hydroxycoumarin.The key targets were GAPDH,VEGFA,STAT3,JUN,HSP90AA1,AKT1,CTNNB1,CASP3,and ALB.Biological process(BP)involved protein phosphorylation,signal transduction,regulation of RNA polymerase II promoter transcription,cell proliferation,apoptosis.Cellular component(CC)involved cytoplasm,nucleus,cell membrane,cellular exosome.Molecular function(MF)involves protein binding,ATPase,threonine kinase,protein kinase activity.KEGG involved cancer pathway,metabolic pathway,PI3K-Akt signalling pathway,cancer proteoglycans,lipids and atherosclerosis,cytomegalovirus infection,microRNAs in cancer,human T-cell leukaemia virus type 1,Ras signalling pathway,MAPK signalling pathway.Molecular docking showed that silverweed lactone H had a strong affinity for each of the other target proteins,indicating that this component plays a key role.The results of RT-qPCR assay and WB assay showed that there were significant differences in gene and protein expression levels before and after drug administration.Conclusion The Chinese medicine in Chloranthus fortunei(A.Gray)Solms-Laub.can treat hepatocellular carcinoma through the MAPK pathway,and the main active ingredients have good docking effects with the core target proteins of the disease.

Objective To understand the current situation of healthcare-associated infection(HAI)in China,pro-vide data support and decision-making basis for formulating scientific and effective strategies for HAI prevention and control.Methods A nationwide cross-sectional survey on HAI was conducted among various types and levels of medical institutions in China according to a unified protocol of bedside surveys and case investigations.Results In 2024,a total of 5 736 medical institutions and 2 751 765 patients were surveyed.Among them,34 889 HAI cases were identified,with a prevalence rate of 1.27％.The number of HAI episodes was 38 032,and case prevalence rate was 1.38％.The prevalence rate of HAI in medical institutions in different regions of China ranged from 0.66％to 2.35％.Among medical institutions of different scales,those with a bed capacity of ≥900 had the high-est incidence of HAI,reaching 1.65％.The most common infection site was the lower respiratory tract(44.66％),followed by the urinary tract(12.94％),surgical site(9.32％),upper respiratory tract(7.02％),and bloodstream infection(5.78％).The top 3 departments with the highest HAI rates were the general intensive care unit(10.02％),department of neurosurgery(5.51％),and department(group)of hematology(5.34％).A total of 23 238 strains of HAI pathogens were detected,with 10 714 strains(46.10％)from lower respiratory tract speci-mens.The top 5 detected strains were Klebsiella pneumoniae(14.76％),Pseudomonas aeruginosa(13.33％),Escherichia coli(12.79％),Acinetobacter baumannii(9.23％),and Staphylococcus aureus(7.88％).231 944 pa-tients underwent class Ⅰ incision surgery were monitored,with 1 647 cases experienced surgical site infection,and the prevalence rate of surgical site infection was 0.71％.The number of patients who should undergo pathogen de-tection(patients receiving therapeutic and therapeutic combined prophylactic antimicrobial agents)was 715 179,while the actual number was 480 492,with a pathogen detection rate of 67.18％.425 225 patients received patho-genic detection before treatment,with a detection rate of 59.46％.Conclusion The overall HAI prevalence in Chi-na is lower,showing disparities among medical institutions of different regions and scales.Therefore,precise imple-mentation of measures is necessary for HAI prevention and control,with a focus on high-risk institutions and high-risk departments,key areas,and critical procedures.All levels of medical institutions should continuously reduce the incidence of HAI by strengthening monitoring,standardizing the use of antimicrobial agents,and reinforcing basic HAI prevention and control measures.

Objective To observe the effects of Yiqi Huoxue Prescription in regulating METTL3/HAND2/TGF-β1 signaling axis on rats with qi deficiency and blood stasis syndrome of coronary heart disease.Methods Totally 48 male SD rats were randomly divided into sham-operation group,model group,Yiqi Huoxue Prescription group and Western medicine(isosorbide mononitrate)group,with 12 rats in each group.Except for the sham-operation group,the animal models of coronary heart disease with qi deficiency and blood stasis syndrome were prepared by the method of ligation of the left anterior descending branch of coronary artery combined with swimming exhaustion.After modeling,Yiqi Huoxue Prescription group and the Western medicine group were gavaged for 28 d.The general conditions of the rats were observed,and electrocardiograms and echocardiograms were collected to detect the cardiac function of the rats,blood rheology was used to detect the degree of blood stasis,HE staining was used to observe the morphology of myocardial tissue,immunohistochemistry was used to detect the expression of METTL3,TGF-β1,Smad2 and Smad3 in myocardial tissue,RT-PCR and Western blot were used to detect the expression of METTL3,HAND2,TGF-β1,Smad2 and Smad3 mRNA and protein in myocardial tissue.Results Compared with the sham-operation group,the model group rats showed symptoms of qi deficiency such as mental lethargy,slow movement,drowsiness,and decreased body mass,significant elevation of ST segment on electrocardiogram,the left ventricular ejection fraction(LVEF)and left ventricular fractional shortening(LVFS)decreased(P＜0.01),the left ventricular end diastolic volume(LVEDV),left ventricular end systolic volume(LVESV),left ventricular end diastolic diameter(LVEDd)and left ventricular end systolic diameter(LVEDs)increased(P＜0.01),the red blood cell aggregation index,Carcasson viscosity and whole blood viscosity increased(P＜0.01);with enlargement and necrosis of myocardial cells,disordered arrangement of myocardial fibers,and infiltration of inflammatory cells into myocardial tissue,the mRNA and protein expressions of METTL3,TGF-β1,Smad2 and Smad3 in myocardial tissue increased(P＜0.05,P＜0.01),while the mRNA and protein expression of HAND2 decreased(P＜0.01).Compared with the model group,the Yiqi Huoxue Prescription group and the Western medicine group showed significant improvement in the mental state of rats,significant ST segment regression on electrocardiogram,LVEF and LVFS increased(P＜0.01),LVEDV,LVESV,LVEDd and LVEDs decreased(P＜0.05,P＜0.01),with red blood cell aggregation index,Carcasson viscosity and whole blood viscosity decreased(P＜0.01);the arrangement of myocardial fibers was neat,the structure and morphology of myocardial cells were improved,the infiltration of inflammatory cells was reduced,the mRNA and protein expressions of METTL3,TGF-β1,Smad2 and Smad3 in myocardial tissue decreased(P＜0.05,P＜0.01),and the mRNA and protein expression of HAND2 increased(P＜0.05,P＜0.01).Conclusion Yiqi Huoxue Prescription can alleviate the pathological damage of myocardial tissue in rats with coronary heart disease of qi deficiency and blood stasis syndrome,improve heart function and inhibit inflammatory response.Its mechanism may be related to the regulation of the METTL3/HAND2/TGF-β1 signaling axis.