ObjectiveTo integrate network pharmacology prediction with multi-level experimental verification methods， and to explore in depth the therapeutic efficacy and potential mechanism of luteolin in treating gout. MethodsDatabases were used to obtain potential pharmacodynamic targets of luteolin. Protein-protein interaction （PPI） network construction and network pharmacology analysis techniques were used to screen key core targets of luteolin in gout treatment. Further biological function enrichment analysis and signaling pathway analysis were performed on these targets. Molecular docking simulation was used to calculate the binding energy between luteolin and potential core targets， clarifying the strength of their interactions. In the in vivo experiment for hyperuricemia， 48 mice were randomly divided into a blank group， a model group， an allopurinol group （5 mg·kg^-1）， and low-dose （10 mg·kg^-1）， medium-dose （30 mg·kg^-1）， and high-dose （90 mg·kg^-1） luteolin groups. For the first three days， the blank and model groups were gavaged with an equal volume of normal saline， while the allopurinol group and luteolin groups were gavaged with corresponding drugs. From day 4 onwards， modeling was performed by intraperitoneal injection at 12：00 daily （normal saline for the blank group， and oxonic acid potassium-hypoxanthine mixture for other groups， with 300 mg·kg^-1 for each group）. Gavage intervention was administered at 18：00 daily （normal saline for the blank/model groups， and corresponding drugs for the treatment groups） until day 7. After sampling， levels of serum uric acid （UA）， alanine aminotransferase （ALT）， and aspartate aminotransferase （AST） were measured. Levels of xanthine oxidase （XO） in the liver and kidney， ATP-binding cassette transporter G2 （ABCG2） and malondialdehyde （MDA） in the kidney， and superoxide dismutase （SOD） in the liver were determined. Renal HE staining was also performed. In the pharmacodynamic study of gouty arthritis， 36 rats were randomly divided into a blank group， a model group， a colchicine group （0.315 mg·kg^-1）， and low-dose （7 mg·kg^-1）， medium-dose （21 mg·kg^-1）， and high-dose （63 mg·kg^-1） luteolin groups. The model was established by vertically injecting 100 µL of 25 g·L^-1 monosodium urate suspension into the posterior lateral aspect of the right ankle joint （the blank group was injected with an equal volume of normal saline）， with repeated injections every two days for reinforcement. From day 2 after modeling， daily gavage administration was performed （normal saline for the blank/model groups， and corresponding drugs for the treatment groups） for a total of 16 days. During the experiment， ankle swelling and pain threshold were measured regularly. After sampling， levels of serum tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β） were determined. Ankle joints were subjected to HE， Masson， and safranin O-fast green staining， and HE staining was also performed on ankle synovial tissue and various organs. Western blot was used to determine the expression levels of key proteins in gout-related signaling pathways. ResultsNetwork pharmacology analysis predicted that luteolin may regulate over 20 core targets， such as XO， ABCG2， nuclear factor erythroid 2-related factor 2 （Nrf2）， and SOD， through acting on signaling pathways including NF-κB， phosphoinositide 3-kinase/protein kinase B （PI3K/Akt）， and ABC transporters， thereby affecting uric acid metabolism and inflammatory responses. In the hyperuricemia model， compared with the blank group， the model group showed significantly increased serum UA level， liver and kidney XO activity， renal ABCG2 expression， and liver SOD activity （P<0.01）. Compared with the model group， the high-dose luteolin group significantly reduced serum UA level （P<0.01）， inhibited liver and kidney XO activity （P<0.01）， and significantly increased renal ABCG2 expression and liver SOD activity （P<0.01）， effectively alleviating renal oxidative stress damage and improving renal histopathological status. In the gouty arthritis model， compared with the blank group， the model group showed significant ankle swelling， decreased pain threshold， and significantly increased levels of IL-6， IL-1β， and TNF-α in serum and synovial tissue （P<0.01）. The high-dose luteolin group significantly reduced ankle swelling， prolonged hot plate pain threshold， effectively decreased the levels of the above inflammatory factors in serum and synovial tissue （P<0.01）， and significantly improved ankle pathological damage， showing good analgesic and anti-inflammatory effects. Western blot results further confirmed that luteolin significantly upregulated Nrf2 protein expression and downregulated XO and nucleotide-binding oligomerization domain （NOD）-like receptor protein 3 （NLRP3） expression in animals. ConclusionLuteolin can improve symptoms of hyperuricemia and gouty arthritis， and its potential mechanism may be related to inhibiting XO activity， increasing ABCG2 and SOD levels， and regulating Nrf2-mediated oxidative stress-related pathways.

OBJECTIVE T o explore the clinical characteristics and risk factors for plastic bronchitis(PB)in the chil-dren with Mycoplasma pneumoniae pneumonia(MPP).METHODS A retrospective case-control study was con-ducted for the medical data of the children with MPP who hospitalized in pediatrics department of Affiliated Hos-pital of Jining Medical College and underwent bronchoscopy and bronchoalveolar lavage from Jan.2023 to Dec.2024.The enrolled children were divided into the PB group and the non-PB group according to the status of complication with PB.The baseline data,clinical characteristics,laboratory test indexes,imaging features,bron-choscopy findings and treatment outcomes were observed and compared between the two groups of children.RESULTS A total of 734 children with MPP were included in the study,131 of whom were assigned as the PB group,and 603 were assigned as the non-PB group.The children were younger[4.83(1.88,7.00)years],the du-ration of fever was longer,the peak temperature was higher[39.50(39.20,39.80)℃],the percentage of compli-cation with pleural effusion was higher(33.59％),the percentage of extrapulmonary organs involved was higher(27.48％),the levels of white blood cells,neutrophils percentage,C-reactive protein(CRP),lactic dehydrogen-ase(LDH),D-dimer(DD)and alanine aminotransferase(ALT)were higher in the PB group than in the non-PB group,and there were significant differences(P＜0.05).There were significant differences in the percentage of mucosal necrosis under bronchoscopy,number of times of treatments assisted by bronchoscopy and length of hospital stay between the two groups(P＜0.05).CONCLUSIONS The MPP children with PB are characterized by younger rage,longer duration of fever,higher peak temperature,higher percentage of complication with pleural effusion,extrapulmonary organs more likely to be involved,more intensive inflammatory reactions and higher percentage of mucosal necrosis under bronchoscopy.Some of the children need to be treated repeatedly with assis-tance of bronchoscopy,and the length of hospital stay is long.

Objective:To establish a genomic nucleic acid mass spectrometry detection platform for allelic risk associated with Alzheimer's disease.Methods:Whole blood samples of 61 patients diagnosed as Alzheimer's disease in the Affiliated Brain Hospital of Guangzhou Medical University from December 28th, 2023 to 31st, March 2024 were collected and deoxynucleic acid (DNA) was extracted, including 22 males and 39 females, aged (67.36 ± 8.18) years old. After screening out 17 risk gene loci in Chinese population, multiplex polymerase chain reaction primers, single-base extension primers and Sanger sequencing primers were designed. Ten samples were used for primer optimization and debugging through Sanger sequencing and time-of-flight mass spectrometry to establish a detection system. The remaining samples were genotyped using a time-of-flight mass spectrometer and verified by Sanger sequencing for accuracy evaluation. Five samples were selected for gradient dilution and then subjected to time-of-flight mass spectrometry detection to evaluate the detection limit. Three clinical samples, one case of Escherichia coli and one case of Staphylococcus aureus genomic DNA samples were selected for cross-reaction research. The anti-interference ability of the detection system was evaluated against hemolysis, chylous substances and conventional anticoagulants in the samples. Two samples, one wild and one homozygous mutation sample with representative peak shapes, were selected to evaluate the anti-interference ability. Four samples containing the common genotypes of all gene loci in the system were selected and repeated 10 times to evaluate the precision.Results:The minimum intensity of single-base extension primers on mass spectrometry is greater than half of the maximum intensity. All 17 risk gene loci screened were successfully typed. The time-of-flight mass spectrometry detection results of 1,037 loci from 61 samples showed that the genotyping detection rate was 100%. The genotypes of the 20 DNA samples were completely consistent with the results of Sanger sequencing, with an accuracy rate of 100%. The mass spectrometry detection results of five samples after gradient dilution indicated that the low detection limit was 5 ng of DNA. The reaction system has a strong anti-interference ability against hemolysis of samples, chylous substances, conventional anticoagulants and DNA cross-contamination. Homologous allele interference and no cross-reaction between the bacterial genome and 17 gene loci do not affect the risk genome detection results. The results of 10 repeated mass spectrometry tests on 4 samples showed that the precision was 100%.Conclusion:The genomic detection system of Alzheimer's disease risk has been successfully established to provide an auxiliary mean for disease diagnosis and risk assessment.

Oxidative stress(OS)and dopaminergic neuron(DN)dysfunction are implicated in the pathogenesis and progression of various neurological disorders.As the primary active component of the traditional Chinese herb Scutellaria baicalensis,baicalin has garnered significant attention due to its potent antioxidant and anti-inflammatory properties.Baicalin exhibits a particular affinity for the dopamine(DA)system,maintaining cerebral DA levels by regulating oxidative stress(OS)-related pathways,suggesting that the DA system serves as the"intracerebral target system"through which it exerts its neuroprotective effects.Nuclear factor erythroid 2-related factor 2(Nrf2),a central transcription factor regulating redox homeostasis,plays a pivotal role in the anti-OS process mediated by baicalin.This systematic review covers the pharmacological effects of baicalin,providing an in-depth mechanistic analysis of the interaction between OS and DN,with a focus on the latest research progress in baicalin-mediated treatment of OS through the Nrf2 signaling pathway in neurological diseases to provide theoretical references for the pharmacological and molecular mechanisms of baicalin's anti-OS modulation of the DA system for the treatment of neurological diseases.

Objective To construct an in vitro research model for studying human liver regeneration based on human pluripotent stem cells(hPSCs)-derived hepatocyte-like organoid(HLO).Methods The hPSCs-derived HLO was obtained by inducing differentiation and the regeneration model after liver injury was constructed by adding acetaminophen(APAP)at fixed time points in HLO culture conditions to simulate acute liver injury.Subse-quently,HLO with catenin/cadherin-associated protein beta 1(CTNNB1)knockout,a key gene regulating liver re-generation,was constructed using CRISPR/Cas9 gene editing technology,and regeneration experiments with APAP injury were performed.HLO as a model for liver regeneration studies was further evaluated by morphological observation,RT-qPCR,Western blot and pathological analysis.Results Morphology evidence as well as expres-sion of marker genes showed that hPSCs-derived HLO was able to initiate a post-injury regeneration response after APAP treatment.CTNNB1-deficient HLO showed delayed recovery in dimension and down-regulated or delayed ex-pression of related genes during post-injury regeneration as compared to control HLO.Conclusions A HLO-based hPSCs-derived human liver regeneration model is successfully constructed,which can be used for gene function studies during liver regeneration.

Objective To investigate alteration of blood routine parameters,blood gas analysis profile,coagulation function,and inflammatory factors during exposure to high-altitude hypoxic environments.Methods Rats were raised in a hypobaric oxygen chamber to simulate the altitude of 5 500 meters.The animals were divided into groups with exposure duration of 0(control),1,3,5,7,14,and 28 days.Arterial blood gas was measured using a blood gas analyzer.The routine blood test was performed by an automatic five-differential animal hematology analyzer.The coagulation function was measured by a fully automatic coagulation analyzer.The level of plasma D-dimer(DD),erythropoietin(EPO),interleukin-10(IL-10),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and C-reactive protein(CRP)was detected by ELISA method.The protein expression of IL-10,IL-6,and TNF-α in lung tissues of the animals was detected by Western blot.Results As compare to control group,the arterial partial pressure of ox-ygen(PaO2)decreased at different durations of hypoxia exposure.The PaO2 in the group exposed to hypoxia for 7 d was the lowest(P＜0.05).The red blood cell count(RBC),hemoglobin(HGB),hematocrit(HCT),mean cor-puscular volume(MCV),mean corpuscular hemoglobin concentration(MCHC)and mean platelet volume(MPV)were all higher than control group(P＜0.05).Erythropoietin(EPO),prothrombin time(PT),activated partial thromboplastin time(APTT),thrombin time(TT),fibrinogen(FIB)and D-dimer(DD)all gradually increased(P＜0.05).The white blood cell count(WBC)and platelet count(PLT),as well as the plasma inflammatory fac-tors including interleukin-10(IL-10),interleukin-6(IL-6),tumor necrosis factor-alpha(TNF-α),C-reactive protein(CRP),and the expression of inflammatory factors in the lung tissue including IL-10,IL-6 and TNF-α pro-teins all showed a curve of increasing at beginning and then followed by a slow decrease with the prolongation of the hypoxia time(P＜0.05).Conclusions In high-altitude hypoxic environment,the blood circulation undergoes dy-namic evolution of functional remodeling with higher risk of inflammatory response.As the hypoxia time prolongs,the animals adapt the environment and the level of inflammatory cytokines gradually decline but remains at a level which is still higher than that of control animals.

AIM:To investigate the regulatory mechanism of silent information regulator 6(Sirt6)on nuclear factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway and ferroptosis in skeletal muscle aging in-duced by D-galactose(D-Gal)in mice.METHODS:A D-Gal-induced mouse aging model was established and randomly divided into control and D-Gal groups.In vitro,D-Gal-treated C2C12 mouse myoblasts were treated with ferroptosis ago-nist erastin(Era)and inhibitor ferrostatin-1(Fer-1),Sirt6 agonist MDL-800 and inhibitor OSS-128167,and Nrf2 siRNA.Mouse body weight and forelimb relative grip strength were monitored.RT-qPCR and Western blot were used to measure the expression of Sirt6,Nrf2,HO-1,P53,P21,P16,muscle ring finger protein 1,muscle atrophy F-box,solute carrier family 7 member 11,and glutathione peroxidase 4 in gastrocnemius muscle and myoblasts.Hematoxylin-eosin staining was performed to examine muscle fiber diameter.Levels of reactive oxygen species(ROS),mitochondrial ROS,mitochon-drial membrane potential,senescence-associated β-galactosidase activity,glutathione,lipid peroxidation,and Fe2? con-centration were measured in myoblasts and myotubes.Immunofluorescence staining was used to detect myosin heavy chain(MyHC)expression in myotubes.RESULTS:Mice in the D-Gal group exhibited significant reductions in body weight and forelimb grip strength(P<0.01),upregulation of aging and muscle atrophy markers,and decreased mRNA and pro-tein levels of ferroptosis markers and Sirt6(P<0.01).Additionally,gastrocnemius muscle fiber diameter significantly de-creased(P<0.01).In D-Gal-treated myoblasts and myotubes,aging and muscle atrophy markers were elevated(P<0.01),MyHC expression was reduced,and protein levels of ferroptosis-related markers,Sirt6,Nrf2,and HO-1 were de-creased(P<0.05 or P<0.01).Fer-1 pre-treatment alleviated these changes(P<0.05 or P<0.01).MDL-800 significantly improved D-Gal-induced aging and muscle atrophy in myoblasts and myotubes,while increasing the expression of ferropto-sis-related proteins(P<0.05 or P<0.01).However,the addition of Erastin abolished the beneficial effects of MDL-800(P<0.05 or P<0.01).Following Nrf2 siRNA transfection,the ability of MDL-800 to improve ferroptosis and the quality of myotube formation was significantly diminished(P<0.05 or P<0.01).CONCLUSION:Sirt6 inhibits ferroptosis in myo-blasts through the Nrf2/HO-1 signaling pathway,thereby alleviating age-related changes in myoblasts and the decline in myotube formation quality,which is beneficial for improving skeletal muscle aging.

Objective In this paper,the ethnic patent medicines(EPMs)in the Chinese Pharmacopeia,National Basic Medical Insurance,Employment Injury Insurance and Maternity Insurance Medicine List,and National Essential Medicine List(re-ferred to as"one pharmacopeia and two lists")were counted and analyzed.Methods The EPMs in"one pharmacopeia and two lists"were summarized,and the information of manufacturers,medicinal materials used in EPMs,and their current quality standards were collated,and their indicative function,adverse reactions,contraindications,and precautions were analyzed.The modern research level of the above EPMs was analyzed by searching relevant literature on CNKI and PubMed.Results A total of 142 EPMs in"one pharmacopeia and two lists"were included,and 68 of the EPMs had exclusive manufacturers and 98.75％of the medicinal materials had provincial-level or higher quality standards.There were issues such as the expression of indicative function in the instruction being mixed between medical terms,91.55％ of the EPMs lacking sufficient information on adverse re-actions,contraindications,and precautions,and modern basic research was weak.Conclusions In the development of EPMs,it is important to improve information on adverse reactions,contraindications,and precautions in the instructions.In expressing indic-ative function,it is necessary to avoid mixing medical terms,which is more conducive to the clinical promotion of EPMs.

AIM:To investigate the immunosuppressive effects of M2-like tumor-associated macrophages(M2-TAMs)on CD8+T cells within the tumor microenvironment of esophageal cancer.METHODS:Multiplex fluores-cence immunohistochemistry was used to analyze the distribution of immune cells in esophageal cancer tissues.An in vitro co-culture system was established,and flow cytometry along with Calcein-AM/PI staining was employed to assess the im-pact of M2-TAMs on CD8+T cell function.The GEPIA database was utilized to evaluate the prognostic significance of PDCD1 expression in esophageal cancer patients and to analyze the correlations between gene expressions.Immunohisto-chemistry(IHC)was performed to detect the expression of TGF-β1 in esophageal cancer tissues.Flow cytometry and en-zyme-linked immunosorbent assay(ELISA)were used to measure PD-1,IFN-γ and TNF-α expression in CD8+T cells fol-lowing treatment with a TGF-β1 inhibitor.RESULTS:Compared with early-stage(stage I)esophageal cancer patients,the patients with advanced disease(stages Ⅱ to Ⅳ)exhibited dynamic changes in the infiltration of CD4+T cells,CD8+T cells,Tregs,and M2-TAMs within tumor tissues,with significant correlations observed among these cell populations(P<0.05).The distribution of M2-TAMs and Tregs was positively correlated with poor prognosis(P<0.05),while that of CD8+T cells was negatively correlated(P<0.05).In contrast,CD4+T cell infiltration showed no significant association with clinical outcomes(P>0.05).Co-culture of CD8+T cells with M2-TAMs resulted in significant downregulation of CD107a,granzyme B,IFN-γ and TNF-α expression(P<0.01).Additionally,M2-TAM-treated CD8+T cells co-cultured with esophageal cancer cells led to reduced apoptosis of cancer cells.High expression of PDCD1 was significantly associated with poor prognosis(P<0.05),and significant correlations were observed between CD8A and PDCD1 expression,as well as between TGF-β1 and CD274 gene expression(P<0.01).TGF-β1 was also significantly associated with CD163+macro-phage infiltration and the progression of esophageal cancer(P<0.05).Treatment with a TGF-β1 inhibitor in the M2-TAM and CD8+T cell co-culture system significantly down-regulated PD-1 expression and increased the secretion of IFN-γ and TNF-α(P<0.01).CONCLUSION:The TGF-β1 derived from M2-TAMs inhibits the antitumor activity of CD8+T cells in the esophageal cancer microenvironment,suggesting potential therapeutic targets for overcoming immunosuppression in esophageal cancer.

Objective:To evaluate the consistency of steady-state blood trough concentration of vancomycin and minimum inhibitory concentration to the area under the serum concentration-time curve (AUC/MIC) and promote the rational use of vancomycin.Methods:It was a single center retrospective study. The clinical data of patients admitted to Xuanwu Hospital, Capital Medical University from January 1, 2019 to December 31, 2020, who were treated with vancomycin and met the inclusion criteria, were collected. Vancomycin calculator was used to calculate AUC/MIC. According to the steady-state blood trough concentration and AUC/MIC standards, the patients were divided into not meeting standard group, meeting standard group, and exceeding standard group, respectively. The standard-reaching status of steady-state blood trough concentration and AUC/MIC in each group was evaluated, as well as the consistency in patients under the 2 different grouping methods. The patients who met the AUC/MIC standard were divided into not meeting standard group, meeting standard group, and exceeding standard group based on steady-state blood trough concentration. The anti-infection efficacy and incidence of vancomycin-associated acute kidney injury (VA-AKI) were evaluated between the different groups to assess clinical effectiveness and safety.Results:A total of 153 patients were included in the study. Among them, 98 (64.1%) patients were male, with age of 61.0 (53.0, 73.0) years, body mass index of 23.9 (21.5, 27.0) kg/m 2, creatinine clearance rate of 107.2 (84.1, 147.0) ml/min, and acute physiology and chronic health evaluationⅡscore of 9.0 (6.0, 14.0) points. Among the 153 patients, the AUC/MIC and steady-state blood trough concentration did not meet standards in 86 cases, met the standards in 17 cases, and exceeded the upper limit of the standards in 14 cases; the results were consistent in 117 cases for both methods, and the consistency rate was 76.5%; the results were inconsistent in 36 patients (23.5%). Among the 36 patients, the steady-state blood trough concentration did not meet the standard in 34 cases [with initial steady-state blood trough concentration of 13.9 (12.8, 14.4) mg/L], while the AUC/MIC met the standard [437 (420, 471)]. AUC/MIC met the standard in 51 patients (33.3%), of which 34 had the steady-state blood trough concentration of less than 15.0 mg/L, and 17 had the concentration of 15.0-20.0 mg/L; the efficacy of anti-infection was 88.2% and 13/17, respectively, the incidence of VA-AKI was 2.9% and 1/17, respectively; the difference was not significant (all P>0.05). No patients had the steady-state blood trough concentration over than 20.0 mg/L. Conclusions:The consistence between steady-state trough concentration of vancomycin and AUC/MIC is 76.5%, which is acceptable. For those who meet the AUC/MIC standard for vancomycin, even if the steady-state blood trough concentration is less than 15.0 mg/L, as long as the AUC/MIC meets the standard, the efficacy and safety can be guaranteed.