Abstract:Objective To investigate the effects of Qingshen Granules(QSG)on folic acid(FA)-induced renal fibrosis in mice and TGF-β1-stimulated HK-2 cells and the underlying mechanism.Methods A mouse model of FA-induced renal fibrosis were treated daily with QSG at 4.0 g·kg-¹ via gavage for 4 weeks.Renal histopathological changes were evaluated using HE and Masson staining,and renal expressions of fibrosis markers,key glycolytic pathway proteins,and histone lactylation-related proteins(Pan Kla,and lactylation sites of H3/H4)were detected using Western blotting.Renal expressions of α-SMA,FN,E-cad,LDHA,and H3K18 lactylation(H3K18la)were also examined with immunofluorescence staining.Serum creatinine(SCR)and blood urea nitrogen(BUN)were measured by ELISA,and renal lactate content was determined with colorimetric assay.In HK-2 cells stimulated with TGF-β1,the effects of QSG-medicated serum from SD rats on cell viability,proliferation,protein expressions of α-SMA,E-cad,LDHA,and H3K18la,extracellular oxygen consumption rate(OCR),extracellular acidification rate(ECAR),and intracellular lactate levels were analyzed.Results The mouse models of renal fibrosis showed increased renal expressions of α-SMA,FN,HIF-1α,HK2,PFKM,PKM2,LDHA,Pan Kla,and H3K18la,elevated serum SCR and BUN and renal lactate level,and decreased E-cad expression.QSG treatment effectively reversed these changes and alleviated renal fibrosis in the mouse models.Immunofluorescence staining showed that QSG treatment significantly reversed the elevation of renal α-SMA,FN and LDHA expressions and reduction of LDHA/H3K18la co-localization,and enhanced E-cad expression.In HK-2 cells,TGF-β1 treatment significantly reduced cell viability,proliferation and OCR,and increased ECAR,lactate,α-SMA,LDHA,and H3K18la,which were all reversed by treatment with QSG-medicated serum.Exogenous lactate obviously reversed the inhibitory effects of QSG on these proteins.Conclusion QSG alleviates renal fibrosis in mice by modulating glycolytic reprogramming,reducing lactate accumulation,and inhibiting H3K18la.

Abstract:Objective To adapt the Liebowitz Social Anxiety Scale for Children and Adolescents(LSAS-CA)to Chinese cultural contexts and assess its reliability and validity among Chinese adolescents.Methods A total of 2103 vocational high school students(917 males and 1186 females;mean age 16.73±1.24 years)in Guangdong Province completed an online survey using the LSAS-CA.Item analysis and psychometric evaluations were conducted.The concurrent validity of the scale was assessed using the Social Anxiety Scale for Adolescents(SAS-A),Patient Health Questionnaire(PHQ-9),and Post-Event Processing Inventory-Trait(PEPI-T),and its discriminant validity was examined using the Rosenberg Self-Esteem Scale(RSES).Three months later,210 of the students were retested to examine the test-retest reliability of LSAS-CA.Results The Chinese LSAS-CA retained 24 items(12 social interaction and 12 performance situations)for assessing anxiety and avoidance with a total of 48 scored responses.Confirmatory factor analysis was used for comparing fitting of 4 competing models(unidimensional,two-factor,bifactor,and higher-order models),and the bifactor model showed the best fit:anxiety bifactor model(χ²=3168.263,df=224,χ²/df=14.144,RMR=0.022,GFI=0.875,AGFI=0.832,PGFI=0.653,NFI=0.931,TLI=0.921,CFI=0.936,and RMSEA=0.079);avoidance bifactor model(χ²=3144.601,df=216,χ²/df=14.558,RMR=0.019,GFI=0.875,AGFI=0.826,PGFI=0.630,NFI=0.944,TLI=0.933,CFI=0.948,and RMSEA=0.080).Conclusion The revised Chinese version of LSAS-CA has acceptable reliability and validity in the context of Chinese culture and provides a more convenient measurement tool for Chinese researchers to study adolescent social anxiety.

Abstract:Objective To investigate the mechanism by which Huangqin Qingre Chubi Capsule(HQC)inhibits synovial angiogenesis mediated by the JAK/STAT pathway in rheumatoid arthritis(RA).Methods An optimized co-culture model of RA-derived fibroblast-like synoviocytes(RA-FLS)and human umbilical vein endothelial cells(HUVECs)was treated with gradient concentrations of HQC-medicated serum with or without plasmid transfection for LncRNA EBLN3P overexpression.The inhibitory effects of HQC on pathological behaviors of RA-FLS were assessed using EdU assay,Transwell invasion assay,and scratch wound healing assay.Cellular secretions of pro-angiogenic factors(VEGF and FGF2)and matrix metalloproteinases(MMP2 and MMP9)were measured by ELISA.HUVEC tube formation capacity and expressions of the endothelial markers CD34 and CD105 were evaluated,and the expressions of molecules in the LncEBLN3P/miR-369-3p/NFIX axis and the JAK2/STAT3 pathway were detected using qRT-PCR and Western blotting.Results The RA-FLS exhibited significantly enhanced proliferation,invasion,and migration with upregulated expressions of VEGF,FGF2,MMP2,and MMP9 and dysregulation of tthe LncEBLN3P/miR-369-3p/NFIX axis,as shown by decreased miR-369-3p and increased expressions of LncEBLN3P,NFIX,JAK2,STAT3,p-JAK2,and p-STAT3.Treatment with HQC-medicated serum effectively reversed these pathological changes,suppressed malignant phenotype of the cells,downregulated angiogenic and matrix-degrading factors,and inhibited the axis and pathway activity.In the LncRNA EBLN3P overexpression(OE-Lnc)model,HQC treatment less effectively reversed the pathological phenotype and pathway activation in the cells as compared to its effect in the non-overexpression setting.Conclusion HQC inhibits pro-angiogenic function of RA-FLS likely by targeting the LncRNA EBLN3P/miR-369-3p/NFIX axis,thereby suppressing the downstream JAK/STAT signaling pathway.

Abstract:Objective To investigate the regulatory role of glutamatergic neurons in the medial prefrontal cortex(mPFC)in the antidepressant effects of esketamine.Methods A total of 150 male C57BL/6J mice were randomly divided into 15 groups(n=10).The mice subjected to the chronic restraint stress(CRS),and those exhibiting depressive-like behaviors following CRS received either esketamine or saline treatment,and subsequent behavioral changes were evaluated.Depressive-like behaviors were assessed using tail suspension test(TST),forced swim test(FST),and sucrose preference test(SPT).The changes in neuronal activity within the mPFC were examined using c-Fos immunofluorescence staining.To explore the underlying mechanism,chemogenetic approaches were employed to specifically modulate the activity of mPFC glutamatergic neurons and examine how these manipulations influenced the effect of esketamine on behaviors of the mice.Results Compared with saline-treated CRS mice,the mice with esketamine treatment showed significantly reduced immobility time in TST and FST and increased sucrose preference rate.Immunofluorescence staining revealed significantly increased expression of c-Fos in glutamatergic neurons in esketamine-treated mice.Chemogenetic activation of mPFC glutamatergic neurons(hM3Dq group)significantly decreased immobility time of the mice in the TST and FST and increased their sucrose preference as compared with the mice in mCherry group(P<0.01),while inhibition of the mPFC glutamatergic neurons(hM4Di group)produced no significant effect.Among the mice treated with esketamine,those in hM4Di group exhibited significantly increased immobility time in TST and FST and decreased sucrose preference compared with those in mCherry group,while the mice in hM3Dq group showed no such changes.Conclusion Esketamine produces antidepressant effects in mice with CRS by activating glutamatergic neurons in the mPFC.

Abstract:Objective To explore the effects of allergens on activation of blood CCR3+granulocyte subtypes in patients with allergic rhinitis(AR).Methods Flow cytometry was used to examine the effects of allergens on CD63 and CD203c expressions on CCR3+CD203c-eosinophils and CCR3+CD203c+basophils of AR patients.Plasma levels of IL-25 and TSLP of the patients were detected with multiplex immunoassay and ELISA,respectively,and their correlations with eosinophils and basophils were analyzed.RNA-sequencing of blood CD16-granulocytes from AR patients was performed,and enrichment analysis was used to explore the functions and potential mechanisms of the differentially expressed genes.Results The percentages of eosinophils and basophils in CCR3+granulocytes increased significantly in AR patients(P<0.005),and exposure to allergens further upregulated the percentage of basophils(P<0.005).CD63 expression was significantly increased in patients with seasonal AR(sAR)and perennial AR(pAR)(P<0.005)but not in AR patients negative for skin prick test(nAR).Allergen exposures enhanced the expressions of CD63 and CD203c on eosinophils and basophils in pAR patients(P<0.005).Elevated plasma IL-25 and TSLP levels in AR patients were associated with increased eosinophil and basophil counts(P<0.005).The 3999 differentially expressed genes in CD16-granulocytes from AR patients positive for skin prick test(SPT+)were involved in protein synthesis,energy metabolism and immune function and in endocytosis,phagosome and carbon metabolism pathways.Conclusion The findings in this study enrich the understanding of the roles of CCR3+eosinophils and basophils in the pathogenesis of SPT+AR and reveal potential therapeutic targets for AR.

Abstract:Objective To investigate the association between cardiovascular health scores(CVH)based on Life's Essential 8(LE8)and 90-day functional prognosis of patients with acute ischemic stroke(AIS)and explore the mediating role of laboratory indicators to provide evidence for secondary stroke prevention.Methods This multicenter,prospective cohort study was conducted among 599 AIS patients within 72 h of onset.The clinical data of the patients were collected,and the cardiovascular health(CVH)scores were calculated according to LE8 criteria.Functional prognosis of the patients at 90 days were assessed by telephone follow-up using the modified Rankin scale score(mRS),whose correlation with the total LE8 score and each individual item score were analyzed using multiple linear regression.Lasso regression analysis was used to select laboratory markers associated with 90-day mRS,and their mediating role in the LE8-outcome relationship were assessed using the Bootstrap method.Results After adjusting for confounders,the total LE8 score was found to inversely correlate with 90-day mRS score of the patients(β=-0.05,95%CI:-0.06‒-0.041;P<0.001).The scores of 7 LE8 components(exercise,diet quality,nicotine exposure,sleep,body mass index,cholesterol,and blood pressure)were inversely correlated with 90-day mRS scores(all P<0.05).Among the laboratory parameters,fibrinogen(FIB)was positively correlated(β=0.215,95%CI:0.068‒0.361;P=0.0041)while MCHC(β=-0.012,95%CI:-0.022‒-0.002;P=0.0229)and HRR(β=-0.007,95%CI:-0.013‒0.00;P=0.042)were negatively correlated with 90-day mRS scores.FIB significantly mediated the relationship between LE8 scores and 90-day mRS scores with an indirect effect of-0.0019(95%CI:-0.00381‒0.00;P=0.006)and a mediation proportion of 3.778%(95%CI:0.00869‒0.08).Conclusion The CVH scores assessed by LE8 are inversely correlated with 90-day functional prognosis of AIS patients,suggesting the importance of maintaining good CVH for improving stroke outcomes.FIB,MCHC and HRR levels are inversely correlated with the patient prognosis,and FIB partially mediates the impact of LE8 scores on stroke outcomes.

Abstract:Objective To investigate the role of the AKT1/GLUT1 signaling pathway in mediating the inhibitory effect of Yiqi Jiedu Formula(YQJDF)against nasopharyngeal carcinoma(NPC)cells.Methods Molecular docking was employed to analyze the binding affinity between the active components of YQJDF and AKT1.MTT assay,real-time cell analysis(RTCA),wound healing assay and Matrigel invasion chamber assay were used to evaluate the effect of YQJDF extract on proliferation,migration and invasion abilities of human NPC cell lines 5-8F and 6-10B,and the changes in cellular protein expression levels were detected using Western blotting.In a BALB/c nude mouse model bearing NPC cell xenografts,tumor growth were observed following treatment with daily gavage with normal saline or YQJDF extract(15.357 g/kg)for 18 consecutive days or with intraperitoneal injections of 5-Fu every other day.The changes in the expressions of AKT1/GLUT1 signaling axis proteins in the xenografts were examined using Western blotting.Results In the NPC cell lines,treatment with YQJDF extract significantly inhibited cell proliferation,migration,and invasion,and downregulated the expressions of p-AKT,GLUT1,XIAP,N-cadherin,and vimentin.The application of GLUT1 or AKT1 activators partially reversed the inhibitory effects of YQJDF on the NPC cells.In the tumor-bearing mouse models,treatment with YQJDF extract obviously suppressed tumor growth and down-regulated the expression of AKT,p-AKT and GLUT1 in the tumor tissues.Conclusion YQJDF inhibits proliferation,invasion,and migration of NPC cells by inhibiting the AKT1/GLUT1 signaling pathway.

Abstract:Objective To investigate the effect of total Astragalus saponins(AST)on astrocyte swelling induced by glucose-oxygen deprivation and reoxygenation(OGD/R)and its molecular mechanism.Methods Primary astrocytes isolated from the brain of post-natal day 2 SD rats were treated with solvent vehicle,AST,or AST combined with U-46619(a p38 activator)24 h prior to and during exposure to OGD/R.The changes in cell volume and morphology were observed,and intracellular reactive oxygen species(ROS)levels and lactate dehydrogenase(LDH)activity were detected.The expression of aquaporin 4(AQP4)on cell membrane was observed using immunofluorescence confocal microscopy,and Western blotting was performed to detect the changes in p38 signaling pathway phosphorylation levels and AQP4 expression level.Results AST treatment significantly reduced OGD/R-induced increase in astrocyte volume,ROS levels and LDH release.The AST-treated cells also exhibited improved cell morphology,reduced aggregation of AQP4 on the cell membrane,and decreased p38 phosphorylation and AQP4 expression levels.The protective effects of AST against OGD/R-induced cell injuries were significantly attenuated by combined treatment of the cells with U-46619.Conclusion AST alleviate astrocyte swelling induced by OGD/R injury by inhibiting the activation of the p38 signaling pathway and AQP4 overexpression on the cell membrane.

Abstract:Objective To investigate the causal relationship between inflammatory proteins and Alzheimer's disease(AD)and the mediating role of plasma metabolites therein.Methods Using Mendelian mandomization(MR)methods and publicly available genome-wide association study(GWAS)data,we selected 91 single nucleotide polymorphisms(SNPs)that were strongly linked to inflammatory proteins without reverse causality with AD as the outcome.A bidirectional two-sample MR analysis was performed.Inflammatory proteins with causal links to AD were identified via inverse variance weighted(IVW)analysis.A mediation MR analysis was then performed using 1400 plasma metabolites to assess their mediating role in this causal pathway.Results The preliminary bidirectional MR analysis identified 3 inflammatory proteins that had a potential positive causal association with AD without reverse causality:Axin-1,C-X-C motif chemokine ligand 11(CXCL11),and interleukin-12β(IL-12β).Elevated levels of Axin-1 were positively causally associated with AD risk(OR=1.082,95%CI:1.009-1.159;P=0.026),while CXCL11(OR=0.951,95%CI:0.914-0.990;P=0.026)and IL-12β(OR=0.959,95%CI:0.926-0.994;P=0.026)were negatively causally associated with AD risk.Sensitivity analyses indicated that these causal effects exhibited no heterogeneity or pleiotropy.In the mediation analysis,18 plasma metabolites with causal links to AD were identified,among which 3 plasma metabolites exhibited mediating effects in the inflammatory protein-AD causal relationship.Methyl-4-hydroxybenzoate sulfate partially mediated the effect between Axin-1 and AD(20.10%).Pregnenetriol sulfate partially mediated the effect between CXCL11 and AD(18.20%).The ratio of spermidine to ornithine played an important mediating role between IL-12β and AD,with a mediating effect of 43.40%.Conclusion This study reveals how specific inflammatory proteins influence AD risk via plasma metabolites and provides genetic evidence for inflammatory-metabolic interactions in AD to facilitate the identification of potential biomarkers and targets for early detection and intervention of AD.

Abstract:Objective To explore the molecular mechanism by which cyclic GMP-AMP synthase-stimulator of interferon genes(cGAS-STING)agonists enhance cytotoxicity of natural killer(NK)cells against gastric cancer cells.Methods NK-92 cells were cultured in X-VIVO15 medium to a density of 1×10⁶/mL and treated with 5,1,or 0.2 μmol/L cGAMP for 24 h before co-culture with gastric cancer MGC-803 and MKN-45 cells in RPMI 1640 medium.The expression of IFN-γ in co-cultured NK-92 cells and tumor cells was detected by qPCR and ELISA.The treated NK-92 cells were then co-cultured with the tumor cells labeled with CellTracker™ CM-DiI at different ratios(1:1,2:1,and 4:1),and tumor cell viability and cytotoxicity of NK-92 cells were determined using a dead cell staining kit and flow cytometry-based cytotoxicity assay.Results The cGAS-STING agonist cGAMP dose-dependently enhanced mRNA expression and secretion of the pro-inflammatory cytokines(such as IFN-γ and TNF-α)in NK-92 cells,and NK-92 cells treated with 5 μmol/L cGAMP showed approximately 3-fold increase of IFN-γ and TNF-α levels.cGAMP treatment also upregulated the expression of activating receptors(such as NKp36 and NKp44)on the cell surface and activated the STING-TBK1-IRF3 signaling pathway in NK-92 cells.NK-92 cells pretreated with cGAMP showed increased cytotoxicity against MGC-803 and MKN-45 cells,which could be reversed by treatment with H-151(a STING inhibitor).In tumor-bearing nude mice,combined treatment with cGAMP and NK cells reduced the mass of xenografts by nearly 40%and significantly slowed tumor growth.Conclusion Activating the cGAS-STING pathway can enhance IFN-γ secretion of NK cells to improve their cytotoxicity against gastric cancer cells,suggesting a therapeutic strategy for gastric cancer using NK cells combined with STING agonists.

Abstract:Objective To investigate the mechanism of XJ-60,a derivative of cajanonic acid A,for alleviating liver fibrosis in mice with non-alcoholic fatty liver disease(NAFLD).Methods Twelve db/db mice were randomized equally into NAFLD model group and XJ-60 treatment group with daily gavage of 50 mg/kg XJ-60,with 6 db/m mice serving as the control group.Successful modeling of NAFLD was validated using HE and Oil Red O staining and by measuring ALT and AST levels of the mice.Immunohistochemistry was used for detecting hepatic expressions of specific protein 1(SP1)and fibronectin(FN);the changes in hepatic expressions of epithelial-mesenchymal transition(EMT)markers,extracellular matrix(ECM)markers,and IL-6 were detected using Western blotting and ELISA.In AML12 cells induced with free fatty acids(FFA),the effect of XJ-60 on expressions of SP1,transforming growth factor-β(TGF-β),Smad3/p-Smad3,and the markers of EMT and ECM were analyzed.SP1 knockdown experiment was performed to validate the role of TGF-β/Smad3 signaling axis in NAFLD.Results XJ-60 treatment significantly reduced serum lipid levels,improved liver histology,and lowered hepatic IL-6 expression in db/db mice.The expression level of SP1 was positively correlated with α-SMA and negatively with E-ca expression.In AML12 cells,XJ-60 treatment at 10 μmol/L significantly reduced FFA-induced lipid accumulation and downregulated cellular expressions of SP1,TGF-β,p-Smad3,and the EMT and ECM markers;SP1 knockdown obviously inhibited the expression levels of TGF-β,Smad3 and ECM markers.Conclusion XJ-60 ameliorates liver fibrosis and steatosis in mice with NAFLD possibly through SP1-mediated inhibition of the TGF-β/Smad3 pathway to reduce ECM deposition.

Abstract:Objective To investigate the protective effect of isovitexin(ISO)on the myocardium of diabetic mice and explore its mechanism.Methods Eighteen adult male C57 mice were randomly divided into control group,diabetes mellitus(DM)group and DM+ISO group(n=6).Primary cultures of neonatal mouse cardiomyocytes(NMCMs)were exposed to high glucose(33 mmol/L)and treated with ISO alone or in combination with 3-TYP(a SIRT3 inhibitor).HE staining was used to evaluate myocardial injury in the diabetic mice,and the levels of MDA,SOD and GSH-Px in the myocardium were detected with ELISA.The expressions of iNOS,NOX2 and 8-OHdG were detected using immunohistochemistry and fluorescence staining.Western blotting was used to analyze the expression levels of NRF2,NOX2,NQO1,and SIRT3,and ROS levels were determined with flow cytometry.Results The diabetic mice showed obvious inflammatory cell infiltration in the myocardium,increased myocardial levels of IL-1β,IL-6 and TNF-α,decreased expression levels of NQO1,NRF2 and SIRT3 proteins,and increased expression levels of NOX2 and AC-SOD2 proteins.ISO treatment of the diabetic mice significantly reduced myocardial inflammatory cell infiltration,lowered the levels of IL-1β,IL-6 and TNF-α,restored the protein levels of NQO1,NRF2 and SIRT3,and decreased the protein levels of NOX2 and AC-SOD2.Conclusion ISO can alleviate diabetic myocardial injury in mice by promoting SIRT3 expression and reducing oxidative stress.

Abstract:Objective To optimize the extraction process of the active components of Radix Paeoniae Alba and assess the antioxidant,anti-inflammatory and pigmentation-reducing effects of Radix Paeoniae Alba extract(BST).Methods Molecular docking identified gallic acid,liquiritin,and paeoniflorin as potential tyrosinase inhibitors,and their contents in BST were determined with high-performance liquid chromatography(HPLC).Based on the contents of the 3 inhibitors,their tyrosinase-inhibiting activity,and DPPH radical scavenging capacity calculated using entropy weight method,the extraction conditions were optimized for solvent ratio,liquid-to-solid ratio,extraction time,and particle size.The inhibitory effects of BST on tyrosinase activity and melanin were assessed in α-MSH-treated B16F10 cells,and its effect on reactive oxygen species(ROS)production was evaluated in H₂O₂-treated HaCaT cells.In a mouse model of melanogenesis induced by UVB radiation,the effect of BST gavage(100,150,and 200 mg/kg)were evaluated by assessing skin conditions of the mice,Fontana-Masson staining,and detecting serum levels of cytokines related to melanogenesis,melanosome transport,metabolism,and inflammation.Results The optimized parameters for BST extraction included 50%ethanol,liquid-to-solid ratio of 15:1,extraction time of 1 h,and 20-mesh particle size.BST exhibited strong free radical-scavenging activity and significantly reduced melanogenesis and increased tyrosinase protein in B16 cells,and lowered ROS production in HaCaT cells.In the mouse models of melanogenesis,BST significantly suppressed the melanogenic enzymes and inflammatory responses by reducing the levels of tyrosinase and TNF-α,thereby effectively reversed UVB-induced photoaging and hyperpigmentation.Conclusion The optimized BST extraction process is stable and predictable.BST shows good skin-whitening effect,which is mediated possibly by inhibiting tyrosinase activity and modulating oxidative stress and inflammatory pathways.

Abstract:Objective To explore the mechanism of Huayu Tongbian Decoction(HYTBD)for relieving slow transit constipation(STC)in rats.Methods Sixty SD rat models of STC induced by intragastric administration of loperamide hydrochloride suspension were randomized equally into 5 groups for treatment with daily gavage with saline(STC model group),low-,medium-,or high-dose HYTBD,or mosapride(positive control group),with another 12 normal rats receiving saline gavage as the blank control group.The first black stool time,fecal water content and intestinal propulsion rate of the rats were measured,and the ultrastructure of interstitial Cajal cells(ICCs)was observed using transmission electron microscopy to calculate the mitochondrial vacuolar rate.The phagocytosis rate of macrophages was detected using immunofluorescence assay,and colonic pathologies were examined using HE staining.The colonic expression levels of c-Kit,CD47,PI3K and Akt were detected with immunohistochemistry or Western Blotting,and their correlations with efferocytosis were analyzed.Results The rat models of STC showed significant ultrastructural damage of the ICCs with increased first black stool time,mitochondrial vacuolar rate,and colonic expression levels of CD47,p-PI3K/PI3K and p-Akt/Akt proteins,lowered fecal water content,intestinal propulsion rate and c-Kit expression level,without significant changes in phagocytic rate.Treatment with HYTBD,especially at the high dose,significantly increased fecal water content,intestinal propulsion rate,phagocytosis rate and colonic c-Kit expression,and decreased the first black stool time,mitochondrial vacuolar rate,and colonic expressions of CD47,p-PI3K/PI3K and p-Akt/Akt in the mouse models.Conclusion HYTBD improves STC in rats possibly by down-regulating colonic CD47 expression and improving macrophage phagocytosis rate via inhibiting PI3K and Akt phosphorylation,thereby promoting efferocytosis of the ICCs.

Abstract:Objective To address the challenges faced by existing artificial intelligence methods in modeling complex heterogeneous biological networks,particularly their limitations in capturing collaborative relationships between nodes and in extracting high-order topological semantic features,we propose a novel drug repositioning prediction method based on dynamic representation learning on heterogeneous graphs.Methods A heterogeneous biological graph that integrates drugs,diseases,and their interaction relationships was constructed,based on which a dynamic gated attention module was designed to extract discriminative topological features of drugs and diseases by incorporating a dynamic graph attention mechanism.A gated residual feature fusion mechanism was developed to precisely integrate structural and semantic information from multiple similarity networks to reduce feature redundancy and information loss,thereby enabling accurate prediction of drug-disease associations.Results Experiments and case studies conducted on multiple drug datasets related to complex diseases demonstrated that the proposed method outperformed existing mainstream models in drug repositioning prediction.Conclusion The proposed method can effectively model complex associations in heterogeneous biological networks,enhance the accuracy of drug repositioning prediction,and provide important technical support for precision treatment of complex diseases and development of medical artificial intelligence.

Abstract:Cardiovascular disease(CVD)is one of the leading causes of death globally,characterized by high morbidity and mortality rates.Metabolic disorders are critical risk factors for CVD.In recent years,growing evidence has highlighted the crucial role of gut microbiota in the onset,progression,and pathological mechanisms of both metabolic diseases and CVD.Akkermansia muciniphila(A.muciniphila),a promising next-generation probiotic,has attracted considerable attention due to its unique metabolic functions and immunomodulatory properties.This review systematically summarizes the mechanisms and research progress regarding the role of A.muciniphila in metabolic diseases and cardiovascular diseases.It elucidates how this bacterium exerts beneficial effects on metabolic disorders through multiple pathways,including improving gut barrier function,regulating lipid and glucose metabolism,increasing short-chain fatty acid production,and suppressing inflammatory responses.Meanwhile,A.muciniphila is also involved in cardiovascular protective processes such as anti-atherosclerosis,blood pressure regulation,alleviation of atrial fibrillation,and improvement of pulmonary arterial hypertension.Its mechanisms involve immunomodulation,regulation of metabolites,and multi-level interactions along the gut-cardiovascular axis,offering novel microbial targets and strategies for intervening in related diseases.

Abstract:Objective To evaluate the synergistic protective effect of linezolid(LZD)combined with Xuebijing Injection(XBJ,a traditional Chinese medicine injection)against pneumonia caused by methicillin-resistant Staphylococcus aureus(MRSA)in mice and explore the underlying mechanism.Methods Fifty C57BL/6J mice with pneumonia induced by tracheal instillation of MRSA were randomized equally into model group,low-and high-dose XBJ treatment groups(13 and 80 mg·kg-1·d-1,respectively),and combined treatment groups with low-and high-dose XBJ(6.5 and 13 mL·kg-1·d-1,respectively)and LZD(80 mg·kg-1·d-1),with 10 PBS-treated mice as the normal control group.The minimum inhibitory concentration(MIC)of XBJ and LZD against MRSA and their effects on bacterial growth and hemolytic activity were assessed in vitro.After the treatments,lung pathologies of the mice were observed with HE staining,and IL-6,TNF-α,IL-1β,and IL-18 levels in lung homogenate were measured using ELISA;the mRNA and protein expressions of Hla,NLRP3,caspase-1,and ASC were detected using qPCR and Western blotting,and pulmonary expressions of NLRP3,caspase-1,and ASC were examined with immunohistochemistry.Results The combined treatment with LZD and XBJ significantly improved survival rates of the mouse models,reduced inflammatory cell infiltration and lung injury scores,decreased the levels of IL-6,TNF-α,IL-1β,and IL-18,and downregulated mRNA and protein expressions of Hla,NLRP3,caspase-1,and ASC.XBJ alone showed no antibacterial activity against MRSA but inhibited α-hemolysin(Hla)secretion both in vitro and in vivo to suppress NLRP3-mediated inflammation,while LZD did not produce this effect.Conclusion LZD combined with XBJ produces enhanced efficacy for improving MRSA pneumonia possibly due to XBJ-induced inhibition of NLRP3-mediated inflammatory response via inhibiting α-hemolysin secretion.

Abstract:Objective To establish a highly sensitive and specific liquid chromatography-tandem mass spectrometry(LC-MS/MS)-based analytical method for detecting steroid hormones in human serum.Methods Solid-phase extraction(SPE)was used for sample pretreatment.Chromatographic conditions were optimized to achieve baseline separation,and mass spectrometry parameters were adjusted to ensure accurate detection of trace amounts of hormones.After validation of the linear range,accuracy,and precision,the method was applied in detection of serum samples from normal pregnant women for analyzing dynamic variation characteristics of 32 steroid hormones during the first,second,and third trimesters of pregnancy.Results The linear range of the method was 0.001-7500 ng/mL,with a limit of detection(LOD)of 0.0007 ng/mL.Both intra-day and inter-day coefficients of variation(CV)of the method were less than 10%,the spiked recovery ranged from 86.21%to 112.67%,the extraction efficiency was 85.05%-106.3%,and the matrix effects were 85.60%-113.22%.The results of detection using this method for serum steroid hormones during pregnancy revealed significant elevation of most of the hormones(such as 11-deoxycortisol,11β-hydroxyprogesterone,dehydroepiandrosterone sulfate,and estriol)with the progression of pregnancy,indicating a close correlation with the development of fetal-placental function.Some hormones increased in the third trimester,possibly due to maternal adaptive regulation and maturation of fetal adrenal function.Conclusion The detection results using the method established in this study reveal dynamic variations of steroid hormones during pregnancy,which provides clues for further physiological and pathological studies of these hormones.

Abstract:Objective To explore the correlation of tea consumption with risks of gastrointestinal diseases using a risk prediction model integrating interpretable machine learning and a large language model.Methods A survey was conducted among the patients undergoing both gastroscopy and 13C-urea breath testing at Gastrointestinal Endoscopy Center of Anxi Hospital of Traditional Chinese Medicine.Univariate analysis was performed to determine the suitability of feature selection.The collected data were randomly divided into training and testing sets in a 7:3 ratio.Support Vector Machine(SVM),K-Nearest Neighbors(KNN),Logistic Regression(LR),Random Forest(RF),Extreme Gradient Boosting(XGB),and Deep Neural Network(DNN)were applied to identify the best classifier for predicting high-risk gastrointestinal conditions.Bayesian optimization algorithm was used to obtain the optimal hyperparameter combinations for the 6 models.After Model fitting,the interpretability of the best models was analyzed using SHapley Additive exPlanations(SHAP).The DeepSeek-R1 base language model was fine-tuned with gastrointestinal disease dataset and Chinese medical online consultation data to obtain the final model.Results The study included 503 participants.All the selected features showed association with gastrointestinal diseases,but only age exhibited a significant linear correlation(β=0.023,SE=0.008,t=2.942,P=0.003).DNN model performed the best with a good accuracy(0.68),precision(0.68),recall rate(0.85),F1 Score(0.75),and AUC(0.74).The top 3 important features were age,DOB value,and smoking history.The large language model constructed provided recommendations consistent with those of professional physicians based on gastroscopy results.Conclusion DNN model is effective for predicting gastrointestinal disease risk and offers reliable support for clinical risk assessment and decision-making regarding endoscopy.Smoking cessation,moderate alcohol consumption,and reasonable tea intake may help prevent gastrointestinal diseases.

Abstract:Objective To explore the mechanism of Xiezhuo Jiedu Recipe(XZJD)for improving ulcerative colitis(UC).Methods SD rat models of UC were randomized into 5 groups(n=10)for daily gavage with saline,mesalamine,or low-,medium-or high-dose XZJD for 14 days,with 10 normal rats with saline gavage as the control group.Pathological changes in the colon of the rats were observed using HE staining,and serum IL-6,IL-10 and IL-17 levels and their expressions in the colon tissues were determined with ELISA and immunohistochemistry.The expressions of occludin and ZO-1 and percentages of Th17 and Treg cells in the colon tissues were detected using immunofluorescence staining and flow cytometry;the mRNA and protein expressions of ROR-γt and Foxp3 were detected with RT-PCR and Western blotting.Results The rat models of UC showed obvious colonic mucosal damage with massive inflammatory cell infiltration,increased IL-6 and IL-17 levels and lowered IL-10 level in both the serum and colonic tissues,reduced expression levels of occludin and ZO-1,increased Th17 and lowered Treg cell percentages,increased RORγt mRNA and protein expressions,and reduced Foxp3 mRNA and protein expressions in the colonic tissue.Treatment with XZJD significantly alleviated inflammatory response in the colonic mucosa and effectively reversed the changes in IL-6,IL-17 and IL-10 levels,expressions of occludin and ZO-1,percentages of Th17 and Treg cells,and the expressions of RORγt and Foxp3 in the colonic tissues of UC rats.Conclusion XZJD can alleviate pathologies in the colonic mucosa in UC rats possibly by regulating Th17/Treg immune balance.