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.

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.

Objective To evaluate the clinical efficacy of haloperidol combined with auricular point pressing therapy in children with tic disorders.Methods A total of 120 pediatric patients diagnosed with tic disorders at the Department of Pediatric Rehabilitation,Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine,Hebei Province from March 2022 to March 2024 were enrolled.Participants were randomly divided into an observation group(n=60)and a control group(n=60)using a random number table.The control group received haloperidol alone,while the observation group received additional auricular point pressing therapy.Treatment duration was 12 weeks.After 3 months,clinical efficacy was assessed by comparing:Yale Global Tic Severity Scale(YGTSS)scores,serum 5-hydroxytryptamine(5-HT)and dopamine(DA)levels,immune function markers(CD3+,CD4+),and incidence of adverse reactions between groups.Results(1)The overall efficacy rate in the observation group was 91.67％(55/60),while that in the control group was 76.67％(46/60).The efficacy of the observation group was superior to that of the control group,with a statistically significant difference(P＜0.05).(2)After treatment,the YGTSS scores of children in both groups was significantly improved(P＜0.05),and the observation group showed a significantly greater improvement in YGTSS scores than the control group,with a statistically significant difference(P＜0.05).(3)After treatment,the levels of 5-HT and DA in both groups of children were significantly improved(P＜0.05),and the observation group showed a significantly greater improvement in 5-HT and DA levels than the control group,with a statistically significant difference(P＜0.05).(4)After treatment,CD3+and CD4+levels were significantly improved in both groups(P＜0.05),and the observation group showed a significantly greater improvement in CD3+and CD4+levels than the control group,with a statistically significant difference(P＜0.05).(5)There was no statistically significant difference in the incidence of adverse reactions between the observation group and the control group,with a statistically significant difference(P＞0.05).Conclusion The combination of haloperidol and auricular point pressing therapy significantly improves clinical symptoms and immune function in children with tic disorders,demonstrating both efficacy and safety.

AIM:To investigate the function and mechanism of the smooth muscle relaxant choline theophylli-nate(CT)in activating primordial follicles in mice.METHODS:Experiments were conducted using in vitro culture of 3-day postpartum(dpp)neonatal SPF-grade female mice,intraperitoneal injection in 3-dpp neonatal mice,and oral adminis-tration in 21-dpp adolescent female mice.The mice were divided into control and CT groups.The ovaries were isolated from 3-dpp neonatal mice for the in vitro culture.Hematoxylin staining was used to count the number of primordial and growing follicles,with 10 mice in each group.qPCR was performed to analyze the expression levels of genes related to fol-licle growth,proliferation,and apoptosis,including growth differentiation factor 9(Gdf9),zona pellucida glycoprotein 3(Zp3),proliferating cell nuclear antigen(PCNA),Ki-67,B-cell lymphoma 2(BCL2),BCL2-associated X protein(BAX),and caspase-3,with 9 mice in each group.Immunofluorescence staining was used to detect the expression of pro-liferation-and apoptosis-related proteins,including PCNA,Ki-67,5-bromo-2'-deoxyuridine(BrdU),cleaved caspase-3,and forkhead box O3a(FOXO3a),with 15 mice in each group.Western blot was performed to measure the expression of DEAD-box helicase 4(DDX4),phosphorylated mammalian target of rapamycin(p-mTOR),and phosphorylated protein kinase B(p-Akt),with 9 mice in each group.Intraperitoneal injection of CT was administered to 3-dpp mice,and follicle counting was performed with 10 mice in each group.Western blotting was used to detect p-mTOR and p-Akt expression,with 9 mice in each group.Immunofluorescence was employed to assess FOXO3a nuclear export,with 15 mice in each group.For the oral administration of CT in drinking water to 21-dpp mice,immunofluorescence and hematoxylin staining was used to count follicles,with 9 mice in each group.RESULTS:Compared with control group,CT treatment signifi-cantly increased the number of growing follicles in mice.The mRNA and/or protein levels of Gdf9,Zp3,Ki-67,PCNA and DDX4 were markedly elevated.Further studies revealed that CT treatment significantly increased p-Akt levels in the ovaries but had no significant effect on p-mTOR levels.The PI3K/Akt inhibitor LY294002 also reversed the choline the-ophyllinate-induced increase in growing follicles.CONCLUSION:Choline theophyllinate promotes the activation of pri-mordial follicles in mice via the PI3K/Akt signaling pathway in oocytes.

To investigate the genetic relatedness between carbapenem-resistant Klebsiella pneumoniae(CRKP) strains isolated from intestinal colonization and subsequent bloodstream infections.

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.

AIM:To investigate the function and mechanism of the smooth muscle relaxant choline theophylli-nate(CT)in activating primordial follicles in mice.METHODS:Experiments were conducted using in vitro culture of 3-day postpartum(dpp)neonatal SPF-grade female mice,intraperitoneal injection in 3-dpp neonatal mice,and oral adminis-tration in 21-dpp adolescent female mice.The mice were divided into control and CT groups.The ovaries were isolated from 3-dpp neonatal mice for the in vitro culture.Hematoxylin staining was used to count the number of primordial and growing follicles,with 10 mice in each group.qPCR was performed to analyze the expression levels of genes related to fol-licle growth,proliferation,and apoptosis,including growth differentiation factor 9(Gdf9),zona pellucida glycoprotein 3(Zp3),proliferating cell nuclear antigen(PCNA),Ki-67,B-cell lymphoma 2(BCL2),BCL2-associated X protein(BAX),and caspase-3,with 9 mice in each group.Immunofluorescence staining was used to detect the expression of pro-liferation-and apoptosis-related proteins,including PCNA,Ki-67,5-bromo-2'-deoxyuridine(BrdU),cleaved caspase-3,and forkhead box O3a(FOXO3a),with 15 mice in each group.Western blot was performed to measure the expression of DEAD-box helicase 4(DDX4),phosphorylated mammalian target of rapamycin(p-mTOR),and phosphorylated protein kinase B(p-Akt),with 9 mice in each group.Intraperitoneal injection of CT was administered to 3-dpp mice,and follicle counting was performed with 10 mice in each group.Western blotting was used to detect p-mTOR and p-Akt expression,with 9 mice in each group.Immunofluorescence was employed to assess FOXO3a nuclear export,with 15 mice in each group.For the oral administration of CT in drinking water to 21-dpp mice,immunofluorescence and hematoxylin staining was used to count follicles,with 9 mice in each group.RESULTS:Compared with control group,CT treatment signifi-cantly increased the number of growing follicles in mice.The mRNA and/or protein levels of Gdf9,Zp3,Ki-67,PCNA and DDX4 were markedly elevated.Further studies revealed that CT treatment significantly increased p-Akt levels in the ovaries but had no significant effect on p-mTOR levels.The PI3K/Akt inhibitor LY294002 also reversed the choline the-ophyllinate-induced increase in growing follicles.CONCLUSION:Choline theophyllinate promotes the activation of pri-mordial follicles in mice via the PI3K/Akt signaling pathway in oocytes.

(±)-Talapyrones A-F (1-6), six pairs of dimeric polyketide enantiomers featuring unusual 6/6/6 and 6/6/6/5 ring systems, were isolated from the fungus Talaromyces adpressus. Their structures were determined by spectroscopic analysis and HR-ESI-MS data, and their absolute configurations were elucidated using a modified Mosher's method and electronic circular dichroism (ECD) calculations. (±)-Talapyrones A-F (1-6) possess a 6/6/6 tricyclic skeleton, presumably formed through a Michael addition reaction between one molecule of α-pyrone derivative and one molecule of C₈ poly-β-keto chain. In addition, compounds 2/3 and 4/5 are two pairs of C-18 epimers, respectively. Putative biosynthetic pathways of 1-6 were discussed.
Polyketides/isolation & purification* ; Talaromyces/chemistry* ; Stereoisomerism ; Molecular Structure ; Circular Dichroism ; Pyrones/chemistry*

Objective: To analyze the factors affecting frailty among convalescent elderly population, so as to provide the evidence for prevention and treatment of frailty. Methods: The convalescent elderly population at ages of 60 years and older were selected from the Wuyunshan Hospital in Hangzhou City using the convenience sampling method from April 2022 to April 2024. Demographic information, chronic disease and medication were collected using questionnaire survey. Frailty was assessed using the FRAIL Scale. Factors affecting frailty among convalescent elderly population were analyzed using a multivariable ordinal logistic regression model. Results: A total of 1 050 questionnaires were distributed, and 1 023 valid questionnaires were collected, with a response rate of 97.43%. There were 793 males (77.52%) and 230 females (22.48%); 192 respondents aged 60 to <70 years (18.77%), 431 respondents aged 70 to <80 years (42.13%) and 400 respondents aged ≥80 years (39.10%); 718 respondents with university degree (70.19%); 890 respondents with a monthly income of 10 000 yuan to <20 000 yuan (87.00%); 130 respondents with comorbidity and polypharmacy (12.71%); and 197 respondents with the risk of malnutrition (19.26%). There were 202 cases with pre-frailty (19.75%) and 47 cases with frailty (4.59%). Multivariable ordinal logistic regression analysis showed that the convalescent elderly population who were aged ≥80 years (OR=3.710, 95%CI: 2.340-5.883), with comorbidity and polypharmacy (OR=12.370, 95%CI: 7.949-20.369) and with the risk of malnutrition (OR=5.414, 95%CI: 3.691-7.933) had higher risk of frailty. Conclusion The high risk of frailty among convalescent elderly population is associated with age, comorbidity and polypharmacy, and malnutrition.

Background and Aims:Preventive temporary stoma has been widely used in surgeries for rectal cancer as a simple and effective method to reduce the severity of postoperative anastomotic leakage.However,some patients with preventive temporary stomas cannot undergo reversal due to various factors,resulting in a permanent stoma.Permanent stomas remain a common adverse outcome in clinical practice,and the reasons behind this are not entirely clear.This study analyzes a continuous surgical sample from a single center to explore the risk factors for forming permanent stoma. Methods:The clinical data of patients who underwent anal-preserving rectal cancer surgery with preventive temporary stoma in Gastrointestinal Cancer Center Ⅲ of Peking University Cancer Hospital from January 2020 to March 2023,with over 12 months of follow-up,were retrospectively collected.The occurrence of permanent stoma was analyzed,and the clinical variables of patients with permanent stoma were compared to those who underwent stoma reversal,along with an analysis of the risk factors for permanent stoma formation.Permanent stoma was defined as ostomy reversal failure for more than 12 months. Results:A total of 299 patients were included,among which 268(89.63％)underwent stoma reversal(stoma closure group),and 31(10.37％)did not(permanent stoma group).Compared to the stoma closure group,the permanent stoma group had a higher incidence of distant organ metastasis at diagnosis(7.5％vs.25.85％,P=0.003)and also had higher proportions of T3 and T4 stages,N2 stage,and clinical stage Ⅳ(all P＜0.05)with an elevated overall postoperative complication rate(19.0％vs.41.9％,P=0.003)as well as a higher rate of severe complications(1.1％vs.9.7％,P=0.016)and an increased incidence of anastomotic leakage(4.9％vs.19.4％,P=0.006).Logistic regression analysis revealed that the presence of distant organ metastasis at diagnosis(OR=5.41,95％CI=1.80-16.27,P=0.003),and occurrence of anastomotic leakage(OR=4.44,95％CI=1.15-17.09,P=0.030)were independent risk factors for the formation of permanent stomas. Conclusion:At present,some patients still cannot undergo reversal of their preventive temporary stoma,resulting in permanent stoma.The formation of permanent stomas is closely related to a low tumor location,distant organ metastasis at diagnosis,and the occurrence of anastomotic leakage.