OBJECTIVES: To investigate the targets and mechanisms of 7-hydroxyethyl chrysin (7-HEC) in prevention and treatment of high-altitude cerebral edema (HACE) in rats. METHODS: Fifty-four male Wistar rats were randomly divided into normal control group, HACE model group, and 7-HEC-treated group (18 rats in each group). Except for the normal control group, rats in the two other groups were exposed to a hypobaric hypoxic chamber simulating a 7000 m altitude for 72 h to establish the HACE model. The 7-HEC-treated group was intraperitoneally injected with 7-HEC (150 mg·kg^－¹·d^－¹) for 3 consecutive days before modeling, while the model group received equivalent isotonic sodium chloride solution. Tandem Mass Tag (TMT) proteomics technology was used to detect differentially expressed proteins (DEPs) with screening criteria set at a fold change >1.2 and P<0.05. Western blotting was used to verify the expression levels of target proteins. Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, and protein-protein interaction (PPI) network analysis were performed. RESULTS: Compared with the normal control group, 256 DEPs were identified in the HACE model group. Compared with the HACE model group, 87 DEPs were identified in the 7-HEC-treated group. Among them, 19 DEPs that were dysregulated in the HACE model group were restored after 7-HEC intervention, of which seven (HSPA4, Arhgap20, SERT, HACL1, CCDC43, POLR3A, and PCBD1) were confirmed by Western blotting. GO enrichment analysis of the DEPs between the HACE model and 7-HEC-treated groups revealed their involvement in 13 biological processes, five cellular components, and two molecular functions. KEGG pathway analysis indicated associations with the mRNA surveillance pathway, Th17 cell differentiation, serotonergic synapse, RNA polymerase, protein processing in the endoplasmic reticulum, peroxisome, neuroactive ligand-receptor interaction, folate biosynthesis. PPI network analysis demonstrated that HSPA4, POLR3A, and HACL1, which were validated by Western blotting, interacted with multiple signaling pathways and ranked among the top 20 hub proteins by degree value, suggesting their potential role as core regulatory factors. Arhgap20, SERT and PCBD1 also exhibited interactions with several proteins, suggesting their potential as key regulatory proteins, whereas no interactions for CCDC43 were identified. CONCLUSIONS This study applied TMT proteomics to identify seven potential therapeutic targets of 7-HEC for the prevention and treatment of HACE. These targets may be involved in the pathogenesis of HACE through multiple pathways, including maintaining cellular homeostasis, ameliorating oxidative stress, regulating energy metabolism, and reducing vascular permeability.
Animals ; Male ; Proteomics/methods* ; Rats, Wistar ; Flavonoids/therapeutic use* ; Rats ; Brain Edema/etiology* ; Altitude Sickness/metabolism* ; Protein Interaction Maps

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 study the therapeutic mechanism of Tuihuang Mixture against cholestasis. METHODS: Forty-eight Wistar rats were randomized equally into blank group, model group, ursodeoxycholic acid group and Tuihuang Mixture group. Except for those in the blank group, all the rats were given α‑naphthylisothiocyanate (ANIT) to establish rat models of cholestasis, followed by treatments with indicated drugs or distilled water. Serum levels of ALT, AST, ALP, γ-GT, TBA and TBIL of the rats were determined, and hepatic expressions IL-1β, IL-18, FXR, NLRP3, ASC, Caspase-1 and GSDMD were detected using q-PCR, ELISA or Western blotting. Histopathological changes of the liver tissues were observed using HE staining. RESULTS: The rat models of cholestasis had significantly increased serum levels of ALT, AST, ALP, γ-GT, TBA and TBIL with increased mRNA and protein expressions of IL-1β and IL-18, decreased protein and mRNA expressions of FXR, and increased protein expressions of NLRP3 and Caspase-1 and mRNA expressions of NLRP3, ASC, Caspase-1 and GSDMD in the liver tissue, showing also irregular arrangement of liver cells, proliferation of bile duct epithelial cells and inflammatory cells infiltration. Treatment of the rat models with Tuihuang Mixture significantly decreased serum levels of ALT, AST, ALP, γ-GT, TBA and TBIL, lowered IL-1β and IL-18 and increased FXR protein and mRNA expressions, and reduced NLRP3, ASC, Caspase-1 and GSDMD proteins and NLRP3, ASC and Caspase-1 mRNA expressions in the liver tissue. Tuihuang Mixture also significantly alleviated hepatocyte injury, bile duct epithelial cell proliferation and inflammatory cell infiltration in the liver of the rat models. CONCLUSIONS Tuihuang Mixture can effectively improve cholestasis in rats possibly by inhibiting NLRP3 inflammatosome-mediated pyroptosis via regulating FXR.
Animals ; NLR Family, Pyrin Domain-Containing 3 Protein ; Rats ; Receptors, Cytoplasmic and Nuclear/metabolism* ; Cholestasis/drug therapy* ; Rats, Wistar ; Inflammasomes/metabolism* ; 1-Naphthylisothiocyanate ; Drugs, Chinese Herbal/therapeutic use* ; Male ; Interleukin-18/metabolism* ; Caspase 1/metabolism* ; Interleukin-1beta/metabolism* ; Liver/metabolism*

As a type of endogenous small non-coding RNA， microRNA （miRNA） can regulate gene expression and thereby intervene against the development and progression of cardiovascular diseases， neurodegenerative diseases， metabolic diseases， and autoimmune diseases. The pathogenesis of cholestasis is complex and is mainly associated with the metabolism and transport of bile acids， oxidative stress， inflammatory response， and intestinal flora. Currently， ursodeoxycholic acid is the preferred drug for the clinical treatment of cholestasis， but it may cause adverse reactions and exhibit poor efficacy in some patients. Studies have shown that miRNA can intervene in the disease process of cholestasis through multiple mechanisms such as regulating bile acid metabolism and transport， alleviating oxidative stress， inhibiting inflammatory response， improving cholangiocyte proliferation， and regulating intestinal flora. It can be used as a new biomarker and action target for cholestasis， with high research potential and value. Therefore， this article summarizes the role and mechanisms of miRNA in regulating cholestasis in recent years， in order to provide a reference for further research on the prevention and treatment of cholestasis by targeting miRNA.

Allergen specific immunotherapy (AIT) is to identify the patient's allergen, give the patient repeated exposure to the allergen extract, and gradually increase the concentration and dose until the target maintenance dose is reached, so that the patient can develop tolerance to the allergen, which is the only treatment that can regulate the pathogenesis of allergic diseases and change its natural course. In recent years, domestic and foreign scholars have made great progress in the clinical practice and research field of AIT. This article reviewed the relevant progress of the mechanism, efficacy and drug administration of AIT.

Objective To study the levels of serum Periostin and interleukin(I1-)-18 in preterm infants with bronchopulmonary dysplasia(BPD),and to analyze their correlation with the severity of the disease and their predictive value for BPD.Methods A total of 62 preterm infants with BPD diagnosed and treated in the hospital from January 2019 to January 2022 were retrospectively selected as the BPD group,and 80 preterm in-fants without BPD during the same period were selected as the non-BPD group.According to the severity of BPD,the infants with BPD were divided into mild subgroup(22 cases),moderate subgroup(24 cases)and se-vere subgroup(16 cases).The serum levels of Periostin and IL-18 were detected by enzyme-linked immu-nosorbent assay.Pearson correlation analysis was used to analyze the correlation between the clinical parameters.Multivariate Logistic regression was used to analyze the influencing factors of BPD,and receiver operating characteristic(ROC)curve was used to analyze the predictive value of each indicator for BPD.Results Compared with the non-BPD group,the BPD group had a significantly higher proportion of infants with pulmonary surfactant(PS)use,neonatal respiratory distress syndrome,apnea,patent ductus arteriosus,and serum levels of Periostin and IL-18,as well as a significantly longer duration of mechanical ventilation,noninvasive respiratory support,and length of hospital stay.The lung function parameters[tidal volume per kilogram(VT/kg),ratio of time to peak tidal expiratory flow to time(TPTEF/TE),ratio of volume to peak tidal expiratory flow to volume(VPEF/VE),expiratory flow at 50％tidal volume(50％TEF),expiratory flow at 75％tidal volume(75％TEF)]and 1,5 min Apgar score of BPD group were lower than that of non-BPD group,and the differences were statistically significant(P＜0.05).The serum levels of Periostin and IL-18 in mild subgroup,moderate subgroup and severe subgroup were increased in turn.The levels of serum Periostin and IL-18 were negatively correlated with pulmonary function indexes(VT/kg,50％TEF,75％TEF,TPTEF/TE,VPEF/VE,P＜0.05).Serum Periostin,IL-18 and neonatal respiratory distress syndrome were independent risk factors for BPD(P＜0.05),and PS was a protective factor(P＜0.05).Serum Periostin,IL-18 and neonatal respiratory distress syndrome were independent risk factors for the severity of BPD(P＜0.05).The area under the curve(95％CI)of serum Periostin and IL-18 alone and in combination for predicting BPD were 0.841(0.814-0.899),0.863(0.820-0.897),0.922(0.878-0.949),respectively.The sensitivity and specificity of combined prediction were 0.902 and 0.825,respectively.The area under the curve of the combination of the two indica-tors for predicting BPD was greater than that of each index alone,and the difference was statistically signifi-cant(Z=5.357,4.894,P＜0.001).Conclusion The levels of serum Periostin and IL-18 are increased in in-fants with BPD,which are related to the severity of BPD and lung function.The combination of serum Perios-tin and IL-18 has a high predictive value for BPD.

Objective:To investigate the prognostic factors of patients with oral squamous cell carcinoma(OSCC)and to establish a nomogram predictive model for the prediction of overall survival(OS)and disease-free survival(DFS)of the patients with OSCC.Methods:A retrospective analysis was performed on clinicopathological data of 361 OSCC patients underwent surgical treatment.Log-rank test and Cox regression model were used to analyze the prognosis of patients with OSCC.The nomogram was established based on the results of multivariate analysis and the consistency index(C index).Receiver operating characteristic curve(ROC)and calibration curve were used to evaluate the prediction ability and accuracy of the nomogram.Results:277 patients(76.73％)were followed up in this study.Univariate and multivariate analysis showed that age,perineural invasion,tumor differentiation and N stage were the inde-pendent risk factors for OS of patients,while age,perineural invasion,tumor differentiation and clinical stage were the independent risk factors for DFS of the patients(all P＜0.05).The C index of the nomogram for OS and DFS of OSCC patients established by this method was 0.741 and 0.763,respectively.ROC and calibration curves showed that the OS and DFS models fitted well with the actual observa-tion results.Conclusion:The OS and DFS nomogram prognostic model of OSCC patients constructed based on age,perineural inva-sion,tumor differentiation,N stage and clinical stage has good predictive ability and accuracy,may help clinicians to accurately assess the prognosis of the patients,and have important significance for the individualized treatment of OSCC patients.

Allergen specific immunotherapy(AIT)is to identify the patient's allergen,give the patient repeated exposure to the allergen extract,and gradually increase the concentration and dose until the target maintenance dose is reached,so that the patient can develop tolerance to the allergen,which is the only treatment that can regulate the pathogenesis of allergic diseases and change its natural course.In recent years,domestic and for-eign scholars have made great progress in the clini-cal practice and research field of AIT.This article re-viewed the relevant progress of the mechanism,ef-ficacy and drug administration of AIT.

ObjectiveTo investigate the impact of early intervention with Yishen Huazhuo prescription （YHP） on the learning and memory of accelerated aging model mice， as well as its underlying mechanism. MethodForty-eight 3-month-old male SAMP8 mice were randomly assigned into four groups， including the model group， low-dose YHP group， high-dose YHP group， and donepezil group. Additionally， 24 SAMR1 mice of the same age were divided into a control group and a YHP treatment control group， each consisting of 12 mice. The YHP groups received YHP at doses of 6.24 g·kg^-1 and 12.48 g·kg^-1， while the donepezil group was treated with donepezil at a dose of 0.65 mg·kg^-1. The model group and control groups were given physiological saline. The mice were gavaged once daily for a duration of four weeks. Spatial learning and memory abilities of mice were assessed using the Morris water maze test. Immunofluorescence staining was employed to evaluate neuronal density as well as expression levels of M1 microglial （MG） polarization marker inducible nitric oxide synthase （iNOS） and M2 MG polarization marker arginase-1 （Arg-1） in the hippocampus region. Enzyme-linked immunosorbent assay （ELISA） was used to measure serum levels of pro-inflammatory factor interleukin 1β （IL-1β） and anti-inflammatory factor transforming growth factor-β₁ （TGF-β₁）. Furthermore， Western blot analysis was conducted to determine expressions of amyloid β peptide1-42 （Aβ_1-42） along with triggering receptor expressed on myeloid cells 2 （TREM2）/nuclear factor kappa B （NF-κB） signaling pathway-related proteins TREM2， phospho （p）-NF-κB p65， and phospho-inhibitory kappa B kinase β （IKKβ） in the hippocampus. ResultCompared with the control group， the model group exhibited a significantly prolonged escape latency （P<0.01）， a significant reduction in neuron-specific nuclear protein （NeuN） expression in the hippocampus， a significant increase in iNOS expression in MG， and a significant decrease in Arg-1 expression. The serum IL-1β content was significantly increased， while the TGF-β₁ content was significantly decreased. Additionally， there was a significant decrease in TREM2 expression in the hippocampus and significant increases in p-NF-κB p65， p-IKKβ， and Aβ_1-42 expressions （P<0.05， P<0.01）. However， no significant changes were observed in escape latency， times of crossing the platform， and hippocampal NeuN expression in the YHP treatment control group. Conversely， iNOS expression in MG as well as the hippocampal p-NF-κB p65， p-IKKβ， and Aβ_1-42 expressions were significantly decreased. Furthermore， TREM2 expression was significantly increased （P<0.05， P<0.01）. In comparison to the model group， the low-dose YHP group showed a significantly shortened escape latency and an increased number of crossing the platform （P<0.05， P<0.01）. In the high-dose YHP group， the escape latency was significantly shortened （P<0.05）. In the low-dose YHP group， high-dose YHP group， the expression of NeuN in the hippocampus was significantly increased， the expression of iNOS in MG was significantly decreased， and the expression of Arg-l was significantly increased. The serum IL-1β content was significantly decreased， while the TGF-β₁ content was significantly increased. Furthermore， the expression of TREM2 in the hippocampus was significantly increased， and the expressions of p-NF-κB p65， p-IKKβ， and Aβ_1-42 were significantly decreased （P<0.01）. ConclusionEarly YHP intervention may promote the transformation of hippocampal MG from M1 to M2 by regulating the TREM2/NF-κB signaling pathway， reduce the release of neuroinflammatory factors， protect hippocampal neurons， and reduce the deposition of Aβ_1-42， and finally delay the occurrence of learning and memory decline in SAMP8 mice.

Objective To determine the role of cathepsin D(CTSD)in cardiovascular events after acute ST-segment elevation myocar-dial infarction(STEMI).Methods A total of 96 patients with STEMI admitted to the Department of Cardiovascular Medicine of Second Hospital Affiliated to Shenyang Medical College from November 2022 to July 2023 were selected as the STEMI group.In addition,20 patients with normal coronary angiography hospitalized during the same period were selected as the control group.Coronary blood was collected from both groups,and the relative expression levels of CTSD in the arterial blood were detected and compared using real-time quantitative polymerase chain reaction.The STEMI group was divided into low,medium,and high CTSD groups based on CTSD expression,and major adverse cardiovascular events(MACE)that occurred within 6 months following percutaneous coronary interven-tion(PCI)were monitored.Results Arterial blood CTSD expression(1.31[1.03-1.75])in the STEMI group was higher than that in the control group(1.02[0.67-1.48])(P＜0.05).Statistically significant differences in platelet count as well as troponin T,N-terminal pro-B natriuretic peptide,and high-density lipoprotein cholesterol levels were observed among the three groups(P＜0.05).The incidence of MACE was significantly higher in the low CTSD group(65％)than in the high CTSD group(10％,P＜0.05).Receiver operating cha-racteristic(ROC)curve analysis showed that the area under the ROC curve for predicting MACE within 6 months after PCI in patients with STEMI was 0.765(95％CI:0.658-0.872,P＜0.001).Conclusion Whole blood CTSD expression level is abnormal in STEMI patient.CTSD may be an indicator for predicting the prognosis of STEMI and,therefore,a drug target for preventing and treating STEMI.