ObjectiveTo explore the mechanism by which Sini San （SNS） inhibits ferroptosis， alleviates inflammation and myocardial injury， and improves myocardial infarction （MI）. MethodsThe active ingredients of SNS were obtained by searching the Traditional Chinese Medicine System Pharmacology Platform （TCMSP） database， its target sites were predicted using the SwissTargetPrediction Database， and the core components were screened out using the CytoNCA plug-in. The targets of MI and ferroptosis were obtained by using GeneCards， Online Mendelian Inheritance in Man （OMIM） database， DrugBank， Therapeutic Target Database （TTD）， FerrDb database and literature review， respectively. The intersection of these targets of SNS-MI-ferroptosis was plotted as a Venn diagram. The protein-protein interaction （PPI） network was constructed using the STRING database， and the visualization graph was prepared using Cytoscape. The core targets were screened out using the CytoNCA plug-in， and the biological functions were clustered by the MCODE plug-in. Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were performed using the David database. Molecular docking was performed using AutoDock and visualized with PyMOL2.5.2. The Kunming mice were randomly divided into the control group， the model group， the SNS group， and the trimetazidine （TMZ） group. The mice were subcutaneously injected with isoprenaline （ISO， 5 mg·kg^-1·d^-1） to establish an MI model. The drug was continuously intervened for 7 days. The ST-segment changes were recorded by electrocardiogram （ECG）， and the tissue morphology changes were observed by hematoxylin-eosin （HE） staining. Cardiomyocyte ferroptosis was investigated by transmission electron microscopy. Serum creatine kinase （CK）， creatine kinase isoenzyme （CK-MB）， lactate dehydrogenase （LDH）， reduced glutathione （GSH）， and malondialdehyde （MDA） levels were detected by biochemical assay. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum levels of interleukin （IL）-6 and 4-hydroxynonenal （4-HNE）. Immunohistochemical staining was employed to detect IL-6 and phosphorylated signal transducer and transcription activator 3 （p-STAT3） in cardiac tissues. Western blot was used to detect STAT3 and p-STAT3 in cardiac tissues. Real-time PCR was used to detect the levels of IL-6， IL-18， solute carrier family 7 member 11 （SLC7A11）， arachidonic acid 15-lipoxygenase （ALOX15）， and glutathione peroxidase 4 （GPx4） in cardiac tissues. ResultsA total of 121 active ingredients of SNS were obtained， and 58 potential targets of SNS in the treatment of MI by regulating ferroptosis were screened. The three protein modules with a score5 were mainly related to the inflammatory response. The GO function was mainly related to inflammation， and KEGG enrichment analysis showed that SNS mainly regulated ferroptosis- and inflammation- related signaling pathways. Molecular docking indicated that the core component had a higher binding force to the target site. Animal experiments confirmed that SNS reduced the level of p-STAT3 （P0.01）， down-regulated the expression of ALOX15 mRNA （P0.01）， up-regulated the level of serum GSH， and the expressions of SLC7A11 and GPx4 mRNA， reduced MDA and 4-HNE levels （P0.05， P0.01）. Additionally， SNS improved the mitochondrial injury induced by cardiomyocyte ferroptosis， reduced the area of MI， alleviated inflammation and myocardial injury， lowered the levels of serum CK， CK-MB， LDH， IL-6， and the mRNA expression levels of IL-16 and IL-18 （P0.05）， and improved ST segment elevation. ConclusionSNS can reduce ISO-induced STAT3 phosphorylation levels， inhibit ferroptosis in cardiomyocytes， alleviate inflammation and myocardial injury， thereby improving MI.

ObjectiveTo explore the mechanism by which Sini San （SNS） inhibits ferroptosis， alleviates inflammation and myocardial injury， and improves myocardial infarction （MI）. MethodsThe active ingredients of SNS were obtained by searching the Traditional Chinese Medicine System Pharmacology Platform （TCMSP） database， its target sites were predicted using the SwissTargetPrediction Database， and the core components were screened out using the CytoNCA plug-in. The targets of MI and ferroptosis were obtained by using GeneCards， Online Mendelian Inheritance in Man （OMIM） database， DrugBank， Therapeutic Target Database （TTD）， FerrDb database and literature review， respectively. The intersection of these targets of SNS-MI-ferroptosis was plotted as a Venn diagram. The protein-protein interaction （PPI） network was constructed using the STRING database， and the visualization graph was prepared using Cytoscape. The core targets were screened out using the CytoNCA plug-in， and the biological functions were clustered by the MCODE plug-in. Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were performed using the David database. Molecular docking was performed using AutoDock and visualized with PyMOL2.5.2. The Kunming mice were randomly divided into the control group， the model group， the SNS group， and the trimetazidine （TMZ） group. The mice were subcutaneously injected with isoprenaline （ISO， 5 mg·kg^-1·d^-1） to establish an MI model. The drug was continuously intervened for 7 days. The ST-segment changes were recorded by electrocardiogram （ECG）， and the tissue morphology changes were observed by hematoxylin-eosin （HE） staining. Cardiomyocyte ferroptosis was investigated by transmission electron microscopy. Serum creatine kinase （CK）， creatine kinase isoenzyme （CK-MB）， lactate dehydrogenase （LDH）， reduced glutathione （GSH）， and malondialdehyde （MDA） levels were detected by biochemical assay. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum levels of interleukin （IL）-6 and 4-hydroxynonenal （4-HNE）. Immunohistochemical staining was employed to detect IL-6 and phosphorylated signal transducer and transcription activator 3 （p-STAT3） in cardiac tissues. Western blot was used to detect STAT3 and p-STAT3 in cardiac tissues. Real-time PCR was used to detect the levels of IL-6， IL-18， solute carrier family 7 member 11 （SLC7A11）， arachidonic acid 15-lipoxygenase （ALOX15）， and glutathione peroxidase 4 （GPx4） in cardiac tissues. ResultsA total of 121 active ingredients of SNS were obtained， and 58 potential targets of SNS in the treatment of MI by regulating ferroptosis were screened. The three protein modules with a score5 were mainly related to the inflammatory response. The GO function was mainly related to inflammation， and KEGG enrichment analysis showed that SNS mainly regulated ferroptosis- and inflammation- related signaling pathways. Molecular docking indicated that the core component had a higher binding force to the target site. Animal experiments confirmed that SNS reduced the level of p-STAT3 （P0.01）， down-regulated the expression of ALOX15 mRNA （P0.01）， up-regulated the level of serum GSH， and the expressions of SLC7A11 and GPx4 mRNA， reduced MDA and 4-HNE levels （P0.05， P0.01）. Additionally， SNS improved the mitochondrial injury induced by cardiomyocyte ferroptosis， reduced the area of MI， alleviated inflammation and myocardial injury， lowered the levels of serum CK， CK-MB， LDH， IL-6， and the mRNA expression levels of IL-16 and IL-18 （P0.05）， and improved ST segment elevation. ConclusionSNS can reduce ISO-induced STAT3 phosphorylation levels， inhibit ferroptosis in cardiomyocytes， alleviate inflammation and myocardial injury， thereby improving MI.

Alzheimer's disease (AD) is a neurodegenerative disease with clinical hallmarks of progressive cognitive impairment. Synergistic effects of the Aβ-Tau cascade reaction are tightly implicated in AD pathology, and microglial NLRP3 inflammasome activation drives neuronal tauopathy. However, the underlying mechanism of how Aβ mediates NLRP3 inflammasome remains unclear. Herein, we determined that oligomeric Aβ (o-Aβ) bound to microglial Kv2.1 and promoted Kv2.1-dependent potassium efflux to activate NLRP3 inflammasome resulting in neuronal tauopathy by using Kv2.1 inhibitor drofenine (Dfe) as a probe. The underlying mechanism has been intensively investigated by assays with Kv2.1 knockdown in vitro (si-Kv2.1) and in vivo (AAV-ePHP-si-Kv2.1). Dfe deprived o-Aβ of its capability to promote microglial NLRP3 inflammasome activation and neuronal Tau hyperphosphorylation by inhibiting the Kv2.1/JNK/NF-κB pathway while improving the cognitive impairment of 5×FAD-AD model mice. Our results have highly addressed that the Kv2.1 channel is required for o-Aβ-driven microglial NLRP3 inflammasome activation and neuronal tauopathy in AD model mice and highlighted that Dfe as a Kv2.1 inhibitor shows potential in the treatment of AD.

Acute kidney injury (AKI) has high morbidity and mortality, but effective clinical drugs and management are lacking. Previous studies have suggested that macrophages play a crucial role in the inflammatory response to AKI and may serve as potential therapeutic targets. Emerging evidence has highlighted the importance of forkhead box protein O1 (FoxO1) in mediating macrophage activation and polarization in various diseases, but the specific mechanisms by which FoxO1 regulates macrophages during AKI remain unclear. The present study aimed to investigate the role of FoxO1 in macrophages in the pathogenesis of AKI. We observed a significant upregulation of FoxO1 in kidney macrophages following ischemia-reperfusion (I/R) injury. Additionally, our findings demonstrated that the administration of FoxO1 inhibitor AS1842856-encapsulated liposome (AS-Lipo), mainly acting on macrophages, effectively mitigated renal injury induced by I/R injury in mice. By generating myeloid-specific FoxO1-knockout mice, we further observed that the deficiency of FoxO1 in myeloid cells protected against I/R injury-induced AKI. Furthermore, our study provided evidence of FoxO1's pivotal role in macrophage chemotaxis, inflammation, and migration. Moreover, the impact of FoxO1 on the regulation of macrophage migration was mediated through RhoA guanine nucleotide exchange factor 1 (ARHGEF1), indicating that ARHGEF1 may serve as a potential intermediary between FoxO1 and the activity of the RhoA pathway. Consequently, our findings propose that FoxO1 plays a crucial role as a mediator and biomarker in the context of AKI. Targeting macrophage FoxO1 pharmacologically could potentially offer a promising therapeutic approach for AKI.

OBJECTIVES: To explore the mechanism by which Pulsatilla saponin D (PSD) inhibits invasion and metastasis of triple-negative breast cancer (TNBC). METHODS: The public databases were used to identify the potential targets of PSD and the invasion and metastasis targets of TNBC to obtain the intersection targets between PSD and TNBC. The "PSD-target-disease" interaction network was constructed and protein-protein interaction (PPI) analysis was performed to obtain the core targets, which were analyzed for KEGG pathway and GO functional enrichment. Molecular docking study of the core targets and PSD was performed, and the therapeutic effect and mechanism of PSD were verified using Transwell assay and Western blotting in cultured TNBC cells. RESULTS: Network pharmacology analysis identified a total of 285 potential PSD targets and 26 drug-disease intersection core targets. GO analysis yielded 175 entries related to the binding of biomolecules (protein, DNA and RNA), enzyme activities, and regulation of gene transcription. KEGG analysis yielded 46 entries involving pathways in cancer, chemical carcinogenesis-receptor activation, microRNAs in cancer, chemical carcinogenesis-reactive oxygen species, PD-L1 expression and PD-1 checkpoint pathway in cancer. Molecular docking showed high binding affinities of PSD to MTOR, HDAC2, ABL1, CDK1, TLR4, TERT, PIK3R1, NFE2L2 and PTPN1. In cultured TNBC cells, treatment with PSD significantly inhibited cell invasion and migration and lowered the expressions of MMP2, MMP9, N-cadherin and the core proteins p-mTOR, ABL1, TERT, PTPN1, HDAC2, PIK3R1, CDK1, TLR4 as well as NFE2L2 expressionin the cell nuclei. CONCLUSIONS The inhibitory effects of PSD on TNBC invasion and metastasis are mediated by multiple targets and pathways.
Humans ; Triple Negative Breast Neoplasms/metabolism* ; Saponins/pharmacology* ; Pulsatilla/chemistry* ; Female ; Molecular Docking Simulation ; Cell Line, Tumor ; Neoplasm Invasiveness ; Protein Interaction Maps ; Neoplasm Metastasis ; Signal Transduction/drug effects* ; Cell Movement/drug effects*

Objective To investigate the molecular mechanism of lipid metabolism disorder in mouse spleen tissues due to high-altitude hypoxia.Methods Ten C57BL/6 male mice were randomly divided into normoxia group(maintained at an altitude of 400 m)and high-altitude hypoxia group(maintained at 4200 m)for 30 days(n=5).Lipidomics and metabolomics analyses of the spleen tissue of the mice were conducted using liquid chromatography-mass spectrometry(LC-MS)to identify the differential metabolites,which were further analyzed by KEGG enrichment and pathway analyses,and the differential genes were screened through transcriptome sequencing.Bioinformatics analysis was conducted to identify the upstream target genes of the differential metabolites in specific metabolic pathways.RT-qPCR and Western blotting were used to detect mRNA expressions of 11β-hydroxysteroid dehydrogenase 1(HSD11B1),steroid 5α reductase 1(SRD5A1),prostaglandin-endoperoxide synthase 1(PTGS1),hematopoietic prostaglandin D synthetase(HPGDS),xanthine dehydrogenase(XDH),purine nucleoside phosphorylase(PNP),hypoxanthine guanine-phosphoribosyltransferase(HPRT)and extracellular 5'-nucleotidase(NT5E)and protein expressions of HSD11B1,SRD5A1,XDH,PNP and HPRT in the mouse spleens.Results We identified a total of 41 differential lipid metabolites in the mouse spleens,and these metabolites and the differential genes were enriched in steroid hormone biosynthesis,arachidonic acid metabolism,and purine metabolism pathways.Compared to the mice kept in normoxic conditions,the mice exposed to high-altitude hypoxia showed significantly upregulated expressions of adrenosterone,androsterone,prostaglandin D2,prostaglandin J2,xanthine,xanthosine,and uric acid in the spleen with also changes in the expression levels of HSD11B1,SRD5A1,PTGS1,HPGDS,XDH,PNP,HPRT,and NT5E.Conclusion High-altitude hypoxia can result in lipid metabolism disorder in mouse spleen tissue by affecting steroid hormone biosynthesis,arachidonic acid metabolism,and purine metabolism pathways.

There are many kinds of zirconia repair materials that have been introduced at home and abroad.Mechanical and aesthetic properties are the important factors for selecting zirconia materials.The process of diagnosis and treatment by dentists and the production by the workers in laboratory affect the final repair effects.To achieve accurate and efficient simulation of dental repair and treatment,effective cooperation between dentists and technicians is required.In this paper,the factors affecting mechanical and aes-thetic properties in the process of material selection,tooth wearing,restoration design and fabrication,concerning zirconia veneer,sin-gle-crown,fixed bridge and edentulous jaw implant fixed repair were discussed and summarized.The common failure reasons were ana-lyzed in order to improve the communication efficiency between dentists and technicians in the process of zirconia repair system and to a-chieve better repair effects.

Objective To investigate the molecular mechanism of lipid metabolism disorder in mouse spleen tissues due to high-altitude hypoxia.Methods Ten C57BL/6 male mice were randomly divided into normoxia group(maintained at an altitude of 400 m)and high-altitude hypoxia group(maintained at 4200 m)for 30 days(n=5).Lipidomics and metabolomics analyses of the spleen tissue of the mice were conducted using liquid chromatography-mass spectrometry(LC-MS)to identify the differential metabolites,which were further analyzed by KEGG enrichment and pathway analyses,and the differential genes were screened through transcriptome sequencing.Bioinformatics analysis was conducted to identify the upstream target genes of the differential metabolites in specific metabolic pathways.RT-qPCR and Western blotting were used to detect mRNA expressions of 11β-hydroxysteroid dehydrogenase 1(HSD11B1),steroid 5α reductase 1(SRD5A1),prostaglandin-endoperoxide synthase 1(PTGS1),hematopoietic prostaglandin D synthetase(HPGDS),xanthine dehydrogenase(XDH),purine nucleoside phosphorylase(PNP),hypoxanthine guanine-phosphoribosyltransferase(HPRT)and extracellular 5'-nucleotidase(NT5E)and protein expressions of HSD11B1,SRD5A1,XDH,PNP and HPRT in the mouse spleens.Results We identified a total of 41 differential lipid metabolites in the mouse spleens,and these metabolites and the differential genes were enriched in steroid hormone biosynthesis,arachidonic acid metabolism,and purine metabolism pathways.Compared to the mice kept in normoxic conditions,the mice exposed to high-altitude hypoxia showed significantly upregulated expressions of adrenosterone,androsterone,prostaglandin D2,prostaglandin J2,xanthine,xanthosine,and uric acid in the spleen with also changes in the expression levels of HSD11B1,SRD5A1,PTGS1,HPGDS,XDH,PNP,HPRT,and NT5E.Conclusion High-altitude hypoxia can result in lipid metabolism disorder in mouse spleen tissue by affecting steroid hormone biosynthesis,arachidonic acid metabolism,and purine metabolism pathways.

Objective:To establish UPLC specific chromatogram method and content determination method of Dryopteridis Crassirhizomatis Rhizoma; To comprehensively evaluate the Dryopteridis Crassirhizomatis Rhizoma from different producing areas.Methods:UPLC method was used to establish the specific chromatogram of 15 batches of Dryopteridis Crassirhizomatis Rhizoma from different producing areas. The quality was evaluated by cluster analysis and partial least square analysis. The content of total flavonoids was determined by ultraviolet spectrophotometry.Results:The established specific chromatogram identified a total of 21 common peaks, and identified 4 components through comparison with control samples. The clustering and partial least squares analysis showed that the samples from Anshan City in Liaoning Province, Yanbian Korean Autonomous Prefecture in Jilin Province, Jiamusi City in Heilongjiang Province, Jilin City in Jilin Province had a certain consistency. The results of total flavone content determination showed that the content of samples from different producing areas was different.Conclusion:The established UPLC characteristic chromatogram and content determination method of total flavone content of Dryopteridis Crassirhizomatis Rhizoma can reflect the quality of Dryopteridis Crassirhizomatis Rhizoma in different places. The quality of samples from different origins is similar, but the contents of the three producing areas fluctuated greatly.

Objective To analyze the effects of nasal endoscopic continuous penetrating suture and nasal packing on discomfort and complications in patients undergoing deflection of nasal septum(DNS)plasty.Methods 116 patients undergoing DNS plasty were enrolled between March 2018 and March 2023,including 69 cases in packing group score(nasal packing)and 47 cases in suture group(continuous penetrating suture).The scores of pain[evaluated by visual analogue scale(VAS)]and epistaxis caused by nasal dressing changes at 3 and 7 d after surgery,subjective discomfort score(headache,dry mouth,epiphora,sleep difficulties,nasal obstruction and nasal pain)at 3 d after surgery,total inspiratory and expiratory resistance before and at 14 d after surgery,and the occurrence of complications at 1 month after surgery were compared between the two groups.Results At 3 d after surgery,VAS of pain and epistaxis score caused by nasal dressing changes were lower in suture group than packing group(P<0.05),and score of subjective discomfort was lower in suture group(P<0.05).At 14 d after surgery,total inspiratory and expiratory resistance were lower in suture group than packing group(P<0.05).The total incidence of postoperative complications was lower in suture group than packing group(P<0.05).Conclusion Compared with nasal packing,nasal endoscopic continuous penetrating suture can reduce pain and bleeding,improve postoperative subjective discomfort and nasal ventilation function,and reduce complications after DNS plasty.