ObjectiveTo investigate the mechanism of action of Kaixinsan in the treatment of Alzheimer's disease （AD） based on network pharmacology， molecular docking， and animal experimental validation. MethodsThe Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） and the Encyclopedia of Traditional Chinese Medicine（ETCM） databases were used to obtain the active ingredients and targets of Kaixinsan. GeneCards， Online Mendelian Inheritance in Man（OMIM）， TTD， PharmGKB， and DrugBank databases were used to obtain the relevant targets of AD. The intersection （common targets） of the active ingredient targets of Kaixinsan and the relevant targets of AD was taken， and the network interaction analysis of the common targets was carried out in the STRING database to construct a protein-protein interaction（PPI） network. The CytoNCA plugin within Cytoscape was used to screen out the core targets， and the Metascape platform was used to perform gene ontology（GO） functional enrichment analysis and Kyoto encyclopedia of genes and genomes（KEGG） pathway enrichment analysis. The “drug-active ingredient-target” interaction network was constructed with the help of Cytoscape 3.8.2， and AutoDock Vina was used for molecular docking. Scopolamine （SCOP） was utilized for modeling and injected intraperitoneally once daily. Thirty-two male C57/BL6 mice were randomly divided into blank control （CON） group （0.9% NaCl， n=8）， model （SCOP） group （3 mg·kg^-1·d^-1， n=8）， positive control group （3 mg·kg^-1·d^-1 of SCOP+3 mg·kg^-1·d^-1 of Donepezil， n=8）， and Kaixinsan group （3 mg·kg^-1·d^-1 of SCOP+6.5 g·kg^-1·d^-1 of Kaixinsan， n=8）. Mice in each group were administered with 0.9% NaCl， Kaixinsan， or Donepezil by gavage twice a day for 14 days. Morris water maze experiment was used to observe the learning memory ability of mice. Hematoxylin-eosin （HE） staining method was used to observe the pathological changes in the CA1 area of the mouse hippocampus. Enzyme linked immunosorbent assay（ELISA） was used to determine the serum acetylcholine （ACh） and acetylcholinesterase （AChE） contents of mice. Western blot method was used to detect the protein expression levels of signal transducer and activator of transcription 3（STAT3） and nuclear transcription factor（NF）-κB p65 in the hippocampus of mice. ResultsA total of 73 active ingredients of Kaixinsan were obtained， and 578 potential targets （common targets） of Kaixinsan for the treatment of AD were screened out. Key active ingredients included kaempferol， gijugliflozin， etc.. Potential core targets were STAT3， NF-κB p65， et al. GO functional enrichment analysis obtained 3 124 biological functions， 254 cellular building blocks， and 461 molecular functions. KEGG pathway enrichment obtained 248 pathways， mainly involving cancer-related pathways， TRP pathway， cyclic adenosine monophosphate（cAMP） pathway， and NF-κB pathway. Molecular docking showed that the binding of the key active ingredients to the target targets was more stable. Morris water maze experiment indicated that Kaixinsan could improve the learning memory ability of SCOP-induced mice. HE staining and ELISA results showed that Kaixinsan had an ameliorating effect on central nerve injury in mice. Western blot test indicated that Kaixinsan had a down-regulating effect on the levels of NF-κB p65 phosphorylation and STAT3 phosphorylation in the hippocampal tissue of mice in the SCOP model. ConclusionKaixinsan can improve the cognitive impairment function in SCOP model mice and may reduce hippocampal neuronal damage and thus play a therapeutic role in the treatment of AD by regulating NF-κB p65， STAT3， and other targets involved in the NF-κB signaling pathway.

Aim To study the therapeutic effect of al-licin on senna-induced diarrhea in mice and to explore the underlying mechanism.Methods Forty-eight C57BL/6J mice were randomly divided into six groups:control,model,loperamide positive control group(2 mg·kg-1),allicin low-dose group(6 mg·kg-1),allicin medium-dose group(12 mg·kg-1)and allicin high-dose group(18 mg·kg-1).Except for the con-trol group,the diarrhea model was induced in the other groups by intragastric administration of senna leaf ex-tract.After drug administration,several diarrhea indi-ces were measured:the rate of loose stools,diarrhea index,accumulated frequency of loose stools at differ-ent time points within 5 hours,and small intestine pro-pelling rate.Serum levels of TNF-α and IL-6 were de-tected by ELISA.Serum NO content was determined u-sing the Griess method.The activities of SOD and CAT,as well as MDA content in the ileum and colon,were measured.The pathological changes and the ex-pression of mRNA related to intestinal barrier proteins in the ileum and colon were evaluated using HE stai-ning and RT-qPCR.Results Allicin improved diar-rhea symptoms in mice induced by senna leaf.It re-duced the rate of loose stools,diarrhea index,cumula-tive number of loose stools in five hours,and the intes-tinal propulsion rate.Allicin also protected the intesti-nal mucosa,decreased serum TNF-α and IL-6 levels,and lowered MDA content in the intestines.It in-creased serum NO levels and enhanced SOD and CAT activities in the intestines.Additionally,allicin upreg-ulated the mRNA expression of AQP1,AQP4,and ZO-1 in intestinal tissues.Conclusions Allicin has a significant therapeutic effect on senna-induced diarrhea in mice.The underlying molecular mechanisms may involve anti-inflammatory and antioxidant effects,in-creased NO content,and upregulation of mRNA ex-pression of aquaporins and tight-junction proteins.

Aim To explore the effects of nuciferine on cognitive behavior and the underlying mechanisms,white matter injury(WMI),neuroinflammation,and ferroptosis in mice with chronic ischemic WMI.Meth-ods Sixty C57BL/6 mice were divided into a control group,a bilateral common carotid artery stenosis(BCAS)model group,and low/high-dose nuciferine groups(20/40 mg·kg-1).A chronic ischemic WMI model was established using BCAS surgery.Following eight weeks of treatment,cognitive behavior(Y-maze,novel object recognition,Morris water maze),white matter integrity(LFB/MBP staining),microglial acti-vation(Iba-1 immunofluorescence),inflammatory cy-tokines(ELISA for TNF-α,IL-1β,IL-6),ferroptosis markers(Fe2+,ROS,MDA,GSH),mitochondrial ultrastructure(electron microscopy),and protein ex-pression of the PI3K/Akt and NRF2/xCT/GPX4 signa-ling pathways(Western blot)were evaluated.Results Compared with the control group,the BCAS group showed significant cognitive decline(P＜0.05),re-duced myelin density,elevated inflammatory cytokines and ferroptosis markers(Fe2+,ROS,MDA),shrunk-en mitochondria,and downregulated PI3K/Akt and NRF2/xCT/GPX4 pathway proteins(P＜0.05).Nu-ciferine intervention significantly ameliorated these in-juries in BCAS mice,with the high-dose group exhibi-ting superior effects(P＜0.05).Conclusions Nu-ciferine exerts protective effects against chronic ische-mic WMI and cognitive impairment by activating the PI3K/Akt and NRF2/xCT/GPX4 signaling pathways,thereby suppressing neuroinflammation and ferroptosis.

AIM To study the effects of Jiaotai Pills and their single composition drugs on high-fat diet-induced hypothalamic inflammation in obese mice.METHODS C57BL/6J mice were randomly divided into the normal group(15 mice)and the high-fat group(75 mice).The mice given 12 weeks of high-fat diet feeding were further randomly divided into the model group,the Jiaotai Pills group,the Coptis chinensis group,the Cinnamomum cassia group and the positive metformin group,with 15 mice in each group.After 6 weeks of administration,the mice had their body weight and fasting blood glucose(FBG)levels detected;their hypothalamic expressions of IL-1β,IL-6,TNF-α and Socs3 mRNA detected by RT-qPCR;their hypothalamic expressions of TLR4,MyD88,IKKβ and activated NF-κB protein detected by Western blot;their hypothalamic expressions of Iba1 and GFAP detected by immunohistochemistry;and their ultrastructural changes of nerve tissues observed using transmission electron microscopy(TEM).RESULTS Compared with the model group,each drug group displayed decreased hypothalamic expressions of IL-1β,IL-6,TNF-α and Socs3 mRNA(P＜0.01),and improved number and morphology of nerve cells revealed by TEM.The groups intervened with Jiaotai Pills,or Coptis chinensis,or metformin shared decreased body weight and FBG levels(P＜0.05);decreased protein expressions of TLR4,MyD88,IKKβ and p-NF-κB(P＜0.05);and decreased number of hypothalamic astrocytes and microglia(P＜0.05).Additionally,decreased p-NF-κB protein expression was observed in the Cinnamomum cassia group(P＜0.05).CONCLUSION Jiaotai Pills and their single composition drugs can improve high-fat diet-induced hypothalamic inflammation in obese mice.

Objective To observe the effect of Kaixin Powder on neuroinflammation in APP/PS1 mice by regulating WDFY1/TLR4/NF-κB signaling pathway;To explore its mechanism of intervening in Alzheimer disease(AD).Methods APP/PS1 transgenic mice were randomly divided into model group,donepezil hydrochloride group(0.66 mg/kg),and Kaixin Powder low-,medium-and high-dosage groups(1.625,3.25,6.5 g/kg),C57BL/6J mice were set as blank control group,with 8 mice in each group,and corresponding drug intervention was given to medicaction group for 24 weeks.Morris water maze,Y maze and novel object recognition experiments were conducted to assess the cognitive function and learning and memory abilities of mice,immunohistochemical staining was used to detect the deposition of β-amyloid protein(Aβ)in hippocampus,the morphology and Nissl bodies of hippocampal CA1 neurons were observed using HE staining and Nissl staining,ELISA was used to detect the serum contents of interleukin(IL)-6,IL-17,IL-1β and tumor necrosis factor-α(TNF-α),Western blot was used to detect the protein expression of calcium-binding adapter molecule 1(Iba1),glial fibrillary acidic protein(GFAP),WDFY1,Toll like receptor 4(TLR4),Toll like receptor associated molecule(TRAM),TIR domain adapter protein(TRIF),NF-κB p65 and p-NF-κB p65 in hippocampal tissue,RT-qPCR was used to detect the mRNA expression of WDFY1,TLR4,TRAM,TRIF and NF-κB p65 in hippocampal tissue.Results Compared with the blank control group,the model group had significantly prolonged escape latency,reduced platform crossings,decreased autonomous reaction alternation rate and relative recognition index(P<0.05,P<0.01),with increased deposition of Aβ in hippocampal tissue(P<0.01),damaged morphological structure of neurons,reduced number of neurons and Nissl bodies,the serum contents of IL-6,IL-17,IL-1β and TNF-α significantly increased,the expression of Iba1,GFAP,WDFY1,TLR4,TRAM,TRIF,p-NF-κB p65 protein and WDFY1,TLR4,TRAM,TRIF mRNA in hippocampal tissue significantly increased(P<0.01).Compared with the model group,Kaixin Powder groups and donepezil hydrochloride group had significantly shortened escape latency and increased platform crossings,autonomous reaction alternation rate and relative recognition index(P<0.05,P<0.01),hippocampal Aβ deposition reduced in Kaixin Powder medium-,high-dosage groups and donepezil hydrochloride group,the morphological structure of neurons recovered,the number of neurons and Nissl bodies increased,the serum contents of IL-6,IL-17,IL-1β and TNF-α significantly decreased(P<0.05,P<0.01),and the protein expression of Iba1,GFAP,WDFY1,TLR4,TRAM,TRIF,p-NF-κB p65 and the mRNA expressions of WDFY1,TLR4,TRAM and TRIF in hippocampal tissue significantly decreased(P<0.05,P<0.01).Conclusion Kaixin Powder can improve cognitive function and learning and memory abilities in AD model mice,alleviate hippocampal neuron damage and Aβ deposition,inhibit the activation of microglia and astrocytes,and thereby reduce serum inflammatory cytokine release.Its mechanism may be related to regulating the WDFY1/TLR4/NF-κB signaling pathway to inhibit neuroinflammation.

BACKGROUND:Pathological cardiac hypertrophy is a risk factor for various heart diseases,but its pathogenesis remains unclear.Circular RNAs are strongly associated with cardiac hypertrophy.However,the role of circular RNA CHACR in cardiac hypertrophy and its regulatory mechanisms have not been clarified.OBJECTIVE:To investigate the role of circular RNA CHACR in pressure overload-induced cardiac hypertrophy and the underlying mechanisms.METHODS:(1)Transverse aortic constriction was used to induce cardiac hypertrophy in vivo after in situ injection of cyclic RNA CHACR overexpressing lentivirus into the heart for 1 week.Heart mass/tibia length ratio and lung mass/tibia length ratio were calculated;cardiomyocyte surface area was measured;hypertrophic marker gene expression levels were detected;myocardial fibrosis degree was detected,and cardiac function was assessed.(2)H9c2 cardiomyocytes were treated with circular RNA CHACR overexpressing lentivirus for 72 hours,and then treated with 1 μmol/L angiotensin Ⅱ for 24 hours to induce hypertrophy of cardiomyocytes.The hypertrophy was assessed by measuring the surface area of cardiomyocytes,the expression level of hypertrophic marker genes,and the protein/DNA ratio.Oxidative stress damage was assessed by detecting reactive oxygen species levels and mitochondrial membrane potential.RESULTS AND CONCLUSION:(1)The expression level of circular RNA CHACR was significantly decreased in both in vivo and in vitro myocardial hypertrophy models(P＜0.01).(2)The overexpression of circular RNA CHACR significantly inhibited the cardiac hypertrophy induced by transverse aortic constriction,including reducing the enlarged heart volume,significantly decreasing the increased heart mass/tibia length ratio(P＜0.05),lung mass/tibia length ratio(P＜0.05),and cardiomyocyte surface area(P＜0.05),and decreasing the upregulated expression levels of hypertrophic markers atrial natriuretic peptide(P＜0.05)and brain natriuretic peptide(P＜0.05).(3)Cardiac fibrosis induced by transverse aortic constriction in mice was significantly inhibited by enforcing expression of circular RNA CHACR,as evidenced by reduced fibrotic area(P＜0.01)and decreased expression levels of the fibrosis marker gene Acta1(P＜0.05).(4)Overexpression of circular RNA CHACR significantly improved cardiac function in mice,including significantly increased ejection fraction(P＜0.05)and fractional shortening(P＜0.01).(5)Enforced expression of circular RNA CHACR significantly inhibited angiotensin Ⅱ-induced cardiomyocyte hypertrophy,including a significant reduction in cardiomyocyte surface area(P＜0.05),downregulation of atrial natriuretic peptide(P＜0.05),and brain natriuretic peptide(P＜0.05)expression levels,and a significant decrease in protein/DNA ratio(P＜0.05).(6)Overexpression of circular RNA CHACR significantly inhibited the elevation of reactive oxygen species levels(P＜0.001)and the decrease in mitochondrial membrane potential(P＜0.05)induced by angiotensin Ⅱ.These results confirm that the expression level of circular RNA CHACR is significantly decreased in cardiac hypertrophy at both in vivo and in vitro myocardial hypertrophy models,and overexpression of circular RNA CHACR significantly inhibits cardiac hypertrophy,alleviates cardiac fibrosis,improves cardiac function,and significantly attenuates angiotensin Ⅱ-induced oxidative stress damage.

Aim To study the therapeutic effect of al-licin on senna-induced diarrhea in mice and to explore the underlying mechanism.Methods Forty-eight C57BL/6J mice were randomly divided into six groups:control,model,loperamide positive control group(2 mg·kg-1),allicin low-dose group(6 mg·kg-1),allicin medium-dose group(12 mg·kg-1)and allicin high-dose group(18 mg·kg-1).Except for the con-trol group,the diarrhea model was induced in the other groups by intragastric administration of senna leaf ex-tract.After drug administration,several diarrhea indi-ces were measured:the rate of loose stools,diarrhea index,accumulated frequency of loose stools at differ-ent time points within 5 hours,and small intestine pro-pelling rate.Serum levels of TNF-α and IL-6 were de-tected by ELISA.Serum NO content was determined u-sing the Griess method.The activities of SOD and CAT,as well as MDA content in the ileum and colon,were measured.The pathological changes and the ex-pression of mRNA related to intestinal barrier proteins in the ileum and colon were evaluated using HE stai-ning and RT-qPCR.Results Allicin improved diar-rhea symptoms in mice induced by senna leaf.It re-duced the rate of loose stools,diarrhea index,cumula-tive number of loose stools in five hours,and the intes-tinal propulsion rate.Allicin also protected the intesti-nal mucosa,decreased serum TNF-α and IL-6 levels,and lowered MDA content in the intestines.It in-creased serum NO levels and enhanced SOD and CAT activities in the intestines.Additionally,allicin upreg-ulated the mRNA expression of AQP1,AQP4,and ZO-1 in intestinal tissues.Conclusions Allicin has a significant therapeutic effect on senna-induced diarrhea in mice.The underlying molecular mechanisms may involve anti-inflammatory and antioxidant effects,in-creased NO content,and upregulation of mRNA ex-pression of aquaporins and tight-junction proteins.

Aim To explore the effects of nuciferine on cognitive behavior and the underlying mechanisms,white matter injury(WMI),neuroinflammation,and ferroptosis in mice with chronic ischemic WMI.Meth-ods Sixty C57BL/6 mice were divided into a control group,a bilateral common carotid artery stenosis(BCAS)model group,and low/high-dose nuciferine groups(20/40 mg·kg-1).A chronic ischemic WMI model was established using BCAS surgery.Following eight weeks of treatment,cognitive behavior(Y-maze,novel object recognition,Morris water maze),white matter integrity(LFB/MBP staining),microglial acti-vation(Iba-1 immunofluorescence),inflammatory cy-tokines(ELISA for TNF-α,IL-1β,IL-6),ferroptosis markers(Fe2+,ROS,MDA,GSH),mitochondrial ultrastructure(electron microscopy),and protein ex-pression of the PI3K/Akt and NRF2/xCT/GPX4 signa-ling pathways(Western blot)were evaluated.Results Compared with the control group,the BCAS group showed significant cognitive decline(P＜0.05),re-duced myelin density,elevated inflammatory cytokines and ferroptosis markers(Fe2+,ROS,MDA),shrunk-en mitochondria,and downregulated PI3K/Akt and NRF2/xCT/GPX4 pathway proteins(P＜0.05).Nu-ciferine intervention significantly ameliorated these in-juries in BCAS mice,with the high-dose group exhibi-ting superior effects(P＜0.05).Conclusions Nu-ciferine exerts protective effects against chronic ische-mic WMI and cognitive impairment by activating the PI3K/Akt and NRF2/xCT/GPX4 signaling pathways,thereby suppressing neuroinflammation and ferroptosis.

Objective To investigate the impact of county-level"Unified ECG Network"construction on the treatment efficiency and clinical outcomes of patients with acute ST-segment elevation myocardial infarction(STEMI).Methods A retrospective analysis was conducted on the clinical data of STEMI patients from Beichuan County and Yanting County in Mianyang City,and Jiange County in Guangyuan City,Sichuan Province,during the 18 months before(128 cases)and 18 months after(187 cases)the establishment of the"Unified ECG Network."Differences in demographic characteristics,treatment efficiency,therapeutic methods,and clinical outcomes between the two groups were compared.Results There was no statistically significant difference in general demographic characteristics between the two groups(all P＞0.05).Compared with the pre-construction group,the post-construction group showed significantly shorter times in initial ECG completion[5(3,7)min vs.6(4,8)min],initial ECG diagnosis[3(2,4)min vs.5(2,6)min],first medical contact to preliminary diagnosis[10(9,12)min vs.13(11,15)min],network hospital door-in-door-out time[21(19,23)min vs.26(23,30)min],and first medical contact to wire-crossing time[(94.82±11.87)min vs.(107.97±18.39)min](allP＜0.001).The proportion of patients bypassing the emergency department and coronary care unit significantly increased(64.17％vs.32.81％,P＜0.001).The proportion of patients undergoing emergency percutaneous coronary intervention significantly increased(72.73％vs.51.56％,P＜0.001),while the proportions of thrombolytic therapy and non-reperfusion therapy significantly decreased(both P＜0.05).Additionally,in-hospital mortality rate,Killip class≥Ⅱ proportion,incidence of major adverse cardiovascular events,and average length of hospital stay were all significantly reduced(all P＜0.05).There were no statistically significant differences among the three county-level chest pain centers in terms of major treatment efficiency,therapeutic strategies,or clinical outcomes(all P＞0.05).Conclusions The construction of the county-level"Unified ECG Network"can significantly improve the treatment efficiency of STEMI patients,optimize reperfusion therapy strategies,improve clinical outcomes,and demonstrate substantial clinical promotion value.

Objective:To investigate the levels of brain natriuretic peptide(BNP),angiotensin Ⅱ(AngⅡ)and Apelin in hypertensive patients with heart failure and their correlation with cardiac function.Methods:A total of 110 pa-tients with hypertension and heart failure(observation group),110 hypertensive patients(hypertension control group)and 100 healthy subjects undergoing physical examination simultaneously(healthy control group)admitted to Chongming Hospital Affiliated to Shanghai University of Medicine & Health Sciences between January 2021 and Oc-tober 2022 were enrolled.BNP,AngⅡ and Apelin levels were compared among above-mentioned groups.Pearson correlation analysis was employed to analyze association of above indexes with cardiac function.Receiver operating characteristic(ROC)curve was used to analyze the diagnostic value of BNP,AngⅡ and Apelin combination for hy-pertension combined heart failure.Results:Compared with participants in healthy control group and hypertension control group,those in observation group had significant higher BNP,AngⅡ,Apelin and left ventricular end-dias-tolic diameter(LVEDd),and significant lower left ventricular ejection fraction(LVEF)(P＜0.001 all).Pearson correlation analysis showed that BNP,AngⅡ,Apelin were negatively correlated with LVEF(r=-0.607,-0.517,-0.549,P＜0.001 all),while they were positively correlated with LVEDd(r=0.695,0.676,0.677,P＜0.001 all).Compared with patients in class Ⅱ and class Ⅲ groups,those in class Ⅳ group had significant higher BNP[(501.42±65.58)pg/ml vs.(382.59±49.69)pg/ml vs.(409.58±53.58)pg/ml],AngⅡ[(3.24±0.84)ng/ml vs.(0.85±0.24)ng/ml vs.(1.06±0.41)ng/ml],Apelin[(6.53±0.71)pg/ml vs.(3.55±0.29)pg/ml vs.(4.98±0.56)pg/ml](P＜0.001 all).ROC curve indicated that the AUC of the combined diagnosis of BNP,AngⅡ and Apelin for hypertensive patients with heart failure of class Ⅲ～Ⅳ was 0.943(95％CI 0.882～0.978),which was significantly higher than that of single index(Z=2.960,6.099,4.653,P＜0.01 all).Conclusion:BNP,AngⅡ and Apelin combination has good performance in diagnosing hypertension complicated heart failure.