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 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.

Objectives:To evaluate the clinical value of the Paris system for reporting urinary cytology (TPS) in the diagnosis of urothelial carcinoma (UC).Methods:A total of 1 744 cytological diagnostic records (from 751 cases) were collected retrospectively. All specimens were voided urines and histopathology as the gold standard. The sensitivity and specificity of urinary cytological diagnosis of UC and risk of high grade malignant (ROHM) in each diagnostic category were compared.Results:There were 360 cases with histopathology. The percentage of negative for high-grade urothelial carcinoma (NHGUC) was 30.1% (226/751), atypical urothelial cells (AUC) was 29.8% (224/751), suspicious for high-grade urothelial carcinoma (SHGUC) was 16.8% (126/751), high grade urothelial carcinoma (HGUC) was 21.2% (159/751), and non-urothelial malignancy (NUM) was 2.1% (16/751). The histpathologic ROHM corresponding to each cytological diagnosis category were 27.3% for NHGUC, 32.7% for AUC, 74.7% for SHGUC, 96.6% for HGUC and 100.0% for NUM, respectively. ROHM of SHGUC was significantly higher than that of AUC group, and the difference between the two groups was statistically significant ( P＜0.001). ROHM of HGUC group was significantly higher than that of SHGUC group, and the difference was statistically significant ( P＜0.001). With SHGUC as the cut-off value, the sensitivity and specificity of cytological diagnosis of HGUC were 76.7% (165/215) and 85.7% (18/21), and with HGUC as the cut-off value, the sensitivity and specificity of cytological diagnosis of HGUC were 53.0% (114/215) and 100.0% (21/21), respectively. Conclusions:Urine cytology has high sensitivity and specificity in the diagnosis of HGUC. The malignant risk of TPS varies with different diagnosis category. The high malignant risk population in cancer hospital leads to the relatively high malignant proportion and ROHM in each diagnosis category. Urinary cytology TPS reporting system is helpful to clinical management and has good clinical application value.

Objective:To prepare 7-hydroxyethyl chrysin(7-HEC)loaded poly(lactic-co-glycolic acid)(PLGA)nanoparticles and to detect the in vitro release.Methods:The 7-HEC/PLGA nanoparticles were prepared by emulsification solvent volatilization method.The particle size,polydispersity index(PDI),encapsulation rate,drug loading and zeta potential were measured.The prescription was optimized by single factor investigation combined with Box-Behnken response surface method.Mannitol was used as protectant to prepare lyophilized powder,and the optimal formulation was characterized and studied for the in vitro release.Results:The optimal formulation of 7-HEC/PLGA nanoparticles was as follows:drug loading ratio of 2.12∶20,oil-water volume ratio of 1∶14.7,and 2.72%soybean phospholipid as emulsifier.With the optimal formulation,the average particle size of 7-HEC/PLGA nanoparticles was(240.28±0.96)nm,the PDI was 0.25±0.69,the encapsulation rate was(75.74±0.80)%,the drug loading capacity was(6.98±0.83)%,and the potentiostatic potential was(-18.17±0.17)mV.The cumulative in vitro release reached more than 50%within 48 h.Conclusions:The optimized formulation is stable and easy to operate.The prepared 7-HEC/PLGA nanoparticles have uniform particle size,high encapsulation rate and significantly higher dissolution rate than 7-HEC.

OBJECTIVE To investigate the protective effect of Tibetan medicine Sanguo decoction on hyperlipidemic rats based on the Nrf2/HO-1 signaling pathway and its related mechanisms. METHODS Forty-eight male SD rats were randomly divided into six groups: the normal control group, the model control group, the simvastatin group(3.5 mg·kg−1), and the Tibetan medicine Sanguo decoction low, medium, and high dose groups(0.43 , 0.86 , 1.72 g·kg−1), with eight rats in each group. The normal control group was fed a basal diet, and the remaining groups were fed the H10060 high-fat diet to prepare a hyperlipidemic rat model. At the same time, each treatment group was given corresponding drugs by gavage once a day. The normal control group and model control group were given an equal volume of physiological saline(once a day) by gavage for 6 consecutive weeks. After 6 weeks, serum levels of lipids[totalcholesterol(TC), triglyceride(TG), low density lipoprotein(LDL) and high density lipoprotein(HDL)] and oxidative parameters[malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione(GSH)] were measured by reagent kit. Nuclear factor erythroid 2-related factor 2(Nrf2), heme oxygenase 1(HO-1), Keap1, and quinone oxidoreductase(NQO1) protein expression in liver tissues were analyzed by Western blotting. The correlation of lipid and oxidative indices was investigated by person correlation. RESULTS Compared with the normal control group, the model control group showed a significant increase in body weight, significantly higher serum levels of TC, TG, LDL, and MDA, significantly lower serum levels of HDL, and significantly lower SOD and GSH activity. Compared with the model control group, each administration group showed a decrease in body weight and serum TC, TG, LDL, and MDA levels. In comparison with the model control group, the body weight was reduced, the serum levels of TC, TG, LDL, and MDA were significantly lower, the serum levels of HDL were significantly higher, and the SOD and GSH activities were significantly higher. Keap1 protein level expression was significantly up-regulated compared with the normal control group, and Nrf2, HO-1, and NQO1 protein level expression were significantly down-regulated in the model control group. Keap1 protein level expression was significantly down-regulated compared to the model control group, and Nrf2, HO-1, and NQO1 protein level expression were significantly up-regulated in the liver tissues of low and high doses of Sanguo decoction. The expressions of Nrf2, HO-1, and NQO1 were significantly up-regulated. Correlation analysis showed that TG was negatively correlated with SOD, HO-1, and NQO1, and positively correlated with Keap1, while TC was negatively correlated with SOD, HO-1, GSH, and Nrf2, and positively correlated with Keap1 and MDA. CONCLUSION The Tibetan medicine Sanguo decoction can improve body weight and blood lipid levels in hyperlipidemic rats, and the mechanism may be related to the regulation of the Nrf2/HO-1 signaling pathway and the improvement of oxidative stress.

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.

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.