ObjectiveThis paper aims to find the identified and validated clinical biomarker data building upon a clinical study of early-phase phase Ⅱ and investigate the correlation analysis of Huanglian Jiedu Wan on syndrome improvement and clinical biomarkers in the treatment of "excess heat-toxicity" based on a machine learning model. Additionally， the effective prediction of clinical biomarker values for the main symptoms of the "excess heat-toxicity" syndrome was assessed. MethodsA total of 229 patients meeting the inclusion criteria for "excess heat-toxicity" syndrome were randomly divided into the Huanglian Jiedu Wan group and the placebo group. Syndrome score transition matrices were constructed for the Huanglian Jiedu Wan group and the placebo group based on three main symptoms of "excess heat-toxicity" syndrome， such as oral ulcers， sore throat， and gum swelling and pain. Data from the patients with these three syndromes were also integrated for an overall analysis. The corresponding syndrome score transition matrices were further constructed to visualize symptom change trends of the patients in the two groups via heatmaps. Based on the identified and validated clinical biomarkers related to inflammation， oxidative stress， and energy metabolism in the early phase， Spearman correlation analysis was employed to analyze and evaluate the associations between clinical biomarkers and syndrome improvement. Key clinical biomarkers reflecting the effect of Huanglian Jiedu Wan were screened through the comparison of differences between groups. An extreme gradient boosting （XGBoost） algorithm was used to develop a prediction model for main symptom classification， with classification performance evaluated through 10-fold cross-validation. Feature importance analysis was applied to identify variables with the greatest contribution to the prediction result. ResultsThe syndrome transition matrix results indicated that the Huanglian Jiedu Wan group showed a superior effect to the placebo group in improving oral ulcers， sore throat， and overall symptoms， with significant effects observed especially in sore throat and overall symptom analyses （P<0.01）. Spearman correlation analysis revealed that several clinical biomarkers positively correlated with "excess heat-toxicity" syndrome and its main symptom improvement， were also called "heat-related biomarkers"， including succinic acid， α-ketoglutaric acid， glycine， lactic acid， adenosine monophosphate （AMP）， tumor necrosis factor-α （TNF-α）， interferon-γ （IFN-γ）， interleukin-1β （IL-1β）， interleukin-4 （IL-4）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， interleukin-10 （IL-10）， and so on. Conversely， clinical biomarkers negatively correlated with symptom severity， were also called "heat-clearing related biomarkers" after administration of Huanglian Jiedu Wan， including malic acid， fumaric acid， cis-aconitic acid， adrenocorticotropic hormone （ACTH）， IL-1β， IL-4， IL-8， succinic acid， and citric acid. The XGBoost classification model using all 52 biomarkers as variables achieved an average test accuracy of 0.754 and an average F1 score of 0.777. Feature importance analysis identified the scores of glutamic acid in saliva and IL-6 were the highest in all the variables， with importance scores of 0.081 and 0.080， respectively. After screening out 14 key variables and optimizing the parameters， model performance improved to an average accuracy of 0.758 and an F1 score of 0.798. Feature importance analysis further determined that the glutamic acid in saliva and IL-6 showed obvious changes after screening the variables， confirming the good syndrome prediction ability of the model constructed by these key clinical biomarkers. ConclusionThis study systematically elucidates the correlation between syndrome improvement and clinical biomarkers of Huanglian Jiedu Wan in the treatment of "excess heat-toxicity" syndrome. An XGBoost classification model based on key clinical biomarkers is successfully established， achieving effective prediction of the symptoms related to the "excess heat-toxicity" syndrome such as oral ulcers and sore throat and providing a new insight for objective identification of traditional Chinese medicine syndromes.

ObjectiveThis paper aims to find the identified and validated clinical biomarker data building upon a clinical study of early-phase phase Ⅱ and investigate the correlation analysis of Huanglian Jiedu Wan on syndrome improvement and clinical biomarkers in the treatment of "excess heat-toxicity" based on a machine learning model. Additionally， the effective prediction of clinical biomarker values for the main symptoms of the "excess heat-toxicity" syndrome was assessed. MethodsA total of 229 patients meeting the inclusion criteria for "excess heat-toxicity" syndrome were randomly divided into the Huanglian Jiedu Wan group and the placebo group. Syndrome score transition matrices were constructed for the Huanglian Jiedu Wan group and the placebo group based on three main symptoms of "excess heat-toxicity" syndrome， such as oral ulcers， sore throat， and gum swelling and pain. Data from the patients with these three syndromes were also integrated for an overall analysis. The corresponding syndrome score transition matrices were further constructed to visualize symptom change trends of the patients in the two groups via heatmaps. Based on the identified and validated clinical biomarkers related to inflammation， oxidative stress， and energy metabolism in the early phase， Spearman correlation analysis was employed to analyze and evaluate the associations between clinical biomarkers and syndrome improvement. Key clinical biomarkers reflecting the effect of Huanglian Jiedu Wan were screened through the comparison of differences between groups. An extreme gradient boosting （XGBoost） algorithm was used to develop a prediction model for main symptom classification， with classification performance evaluated through 10-fold cross-validation. Feature importance analysis was applied to identify variables with the greatest contribution to the prediction result. ResultsThe syndrome transition matrix results indicated that the Huanglian Jiedu Wan group showed a superior effect to the placebo group in improving oral ulcers， sore throat， and overall symptoms， with significant effects observed especially in sore throat and overall symptom analyses （P<0.01）. Spearman correlation analysis revealed that several clinical biomarkers positively correlated with "excess heat-toxicity" syndrome and its main symptom improvement， were also called "heat-related biomarkers"， including succinic acid， α-ketoglutaric acid， glycine， lactic acid， adenosine monophosphate （AMP）， tumor necrosis factor-α （TNF-α）， interferon-γ （IFN-γ）， interleukin-1β （IL-1β）， interleukin-4 （IL-4）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， interleukin-10 （IL-10）， and so on. Conversely， clinical biomarkers negatively correlated with symptom severity， were also called "heat-clearing related biomarkers" after administration of Huanglian Jiedu Wan， including malic acid， fumaric acid， cis-aconitic acid， adrenocorticotropic hormone （ACTH）， IL-1β， IL-4， IL-8， succinic acid， and citric acid. The XGBoost classification model using all 52 biomarkers as variables achieved an average test accuracy of 0.754 and an average F1 score of 0.777. Feature importance analysis identified the scores of glutamic acid in saliva and IL-6 were the highest in all the variables， with importance scores of 0.081 and 0.080， respectively. After screening out 14 key variables and optimizing the parameters， model performance improved to an average accuracy of 0.758 and an F1 score of 0.798. Feature importance analysis further determined that the glutamic acid in saliva and IL-6 showed obvious changes after screening the variables， confirming the good syndrome prediction ability of the model constructed by these key clinical biomarkers. ConclusionThis study systematically elucidates the correlation between syndrome improvement and clinical biomarkers of Huanglian Jiedu Wan in the treatment of "excess heat-toxicity" syndrome. An XGBoost classification model based on key clinical biomarkers is successfully established， achieving effective prediction of the symptoms related to the "excess heat-toxicity" syndrome such as oral ulcers and sore throat and providing a new insight for objective identification of traditional Chinese medicine syndromes.

Objective To investigate the expression of microRNA-221-3p(miR-221-3p)in breast cancer cells under hypoxia conditions,and the effects of knocking down its expression on the stemness maintenance and angiogenesis in breast cancer cells.Methods Human breast cancer cell line MCF-7 were cultured under normoxic and hypoxic conditions,and the expression of miR-221-3p in cells was detected by quantitative real-time polymerase chain reaction(RT-qPCR).MCF-7 cells were divided into normoxic group(MCF-7 cells were cultured under normoxic conditions),hypoxic group(MCF-7 cells were cultured under hypoxic conditions),hypoxia+miR-221-3p NC group(MCF-7 cells were cultured under hypoxic conditions and transfected with miR-221-3p NC),hypoxia+miR-221-3p inhibitor group(MCF-7 cells were cultured under hypoxic conditions and transfected with miR-221-3p inhibitor).The proliferation activity of cells in each group was detected by CCK-8 method;the number of cell tumor spheres in each group was detected by tumor sphere formation assay;the proportion of cells labeled with the stemness marker aldehyde dehydrogenase 1(ALDH1)in each group was detected by flow cytometry;the expression of cluster of differentiation 133(CD133),cluster of differentiation 44(CD44)and sex determination region Y-box protein 2(SOX2)of cells in each group was detected by Western blot;the number of human umbilical vein endothelial cells(HUVEC)tubes formed by cell supernatant culture in each group was detected by in vitro tube formation experiment;the expression of hypoxia-inducible factor-1α(HIF-1α)and vascular endothelial growth factor(VEGF)of cells in each group was detected by Western blot.Results Compared with normoxic group,the expression levels of miR-221-3p,and CD133,CD44,SOX2,HIF-1α,VEGF protein of cells in hypoxic group were significantly up-regulated(P<0.05),and the cell proliferation activity,the number of tumor spheres,the proportion of ALDH1+cells and the number of HUVEC tubes were significantly increased(P<0.05).Compared with hypoxia+miR-221-3p NC group,the expression levels of miR-221-3p,and CD133,CD44,SOX2,HIF-1α,VEGF protein of cells in hypoxia+miR-221-3p inhibitor group were significantly down-regulated(P<0.05),and the cell proliferation activity,the number of tumor spheres,the proportion of ALDH1+cells,and the number of HUVEC were significantly decreased(P<0.05).Conclusion Under hypoxic conditions,the expression of miR-221-3p in breast cancer cells is up-regulated,and knocking down the expression of miR-221-3p can reduce the stemness and angiogenesis ability of breast cancer cells.

BACKGROUND:Lijin manipulation can reduce fibrosis scar hyperplasia and promote skeletal muscle repair.However,improper activation of the Wnt/β-catenin signaling pathway can aggravate the fibrosis of injured skeletal muscle and adversely affect the repair process of skeletal muscle.To study the regulatory effect of Lijin manipulation on the Wnt/β-catenin signaling pathway is conducive to elucidate the related mechanisms of Lijin manipulation in reducing fibrosis scar hyperplasia and promoting skeletal muscle injury repair.OBJECTIVE:To explore the mechanism of Lijin manipulation in promoting the repair of skeletal muscle injury in rabbits.METHODS:Forty-five healthy adult Japanese white rabbits were randomly divided into blank group,model group and Lijin group with 15 rabbits in each group.Gastrocnemius muscle percussion modeling was performed in both model group and Lijin group.Lijin manipulation was performed in the Lijin group on the 3rd day after modeling,once a day,15 minutes once.Five animals in each group were selected and killed on the 7th,14th and 21st days after modeling.The general morphological structure of gastrocnemius was observed by hematoxylin-eosin staining and the content of collagen fiber was observed by Masson staining.Western blot was used to detect the protein expression of Wnt3a,β-catenin,GSK3β,p-GSK3β,TCF,type I collagen and type III collagen in gastrocnemius muscle,and RT-PCR was used to detect the mRNA expression of Wnt3a,β-catenin and TCF.The expression of β-catenin was detected by immunofluorescence,and the expression of type I collagen and type III collagen was detected by immunohistochemistry.RESULTS AND CONCLUSION:The results of hematoxylin-eosin staining and Masson staining showed that compared with the model group,inflammatory cell infiltration and collagen fiber amount decreased in the Lijin group(P<0.001),and muscle fibers gradually healed.Western blot results showed that compared with the model group,the protein expression levels of Wnt3a,β-catenin,TCF,type I collagen and type III collagen were significantly decreased in the Lijin group at all observation time points(P<0.05),while the ratio of P-GSK3β/GSK3β was significantly increased in the Lijin group at all observation time points compared with the model group(P<0.05).RT-PCR results showed that compared with the model group,the mRNA expression levels of Wnt3a,β-catenin and TCF were significantly decreased in the Lijin group at all observation time points(P<0.001).Immunofluorescence results showed that compared with the model group,the fluorescence intensity of β-catenin expression in the Lijin group was significantly decreased at each observation time point and gradually became similar to that in the blank group(P<0.001).Immunohistochemical results showed that the expression levels of type I collagen and type III collagen in the Lijin group were significantly lower than those in the model group(P<0.01).To conclude,Lijin manipulation could inhibit the abnormal activation of the Wnt/β-catenin signaling pathway,reduce fibrotic scar hyperplasia,and promote the repair of injured skeletal muscle.

BACKGROUND:Excessive apoptosis in skeletal muscle cells will destroy the dynamic balance of the number of myocytes,leading to pathological injury of skeletal muscle.Lijin manipulation is effective in treating skeletal muscle injury,but whether it can inhibit apoptosis and promote the repair of skeletal muscle injury is unknown.OBJECTIVE:To explore the mechanism by which Lijin manipulation reduces inflammation and apoptosis during the repair of skeletal muscle injury in rabbits.METHODS:Forty-five healthy adult Japanese white rabbits were randomly divided into blank group,model group and Lijin group(n=15 per group).No intervention was performed in the blank group.Gastrocnemius muscle percussion molding was performed in both the model group and Lijin group.After modeling,the model group was not treated,while the Lijin manipulation(Stroking,kneading,and rubbing)was performed in the Lijin group on the 3rd day,once a day,15 min/time.Sampling in each group was performed on the 7th,14th and 21st days after modeling.The general morphological structure of gastrocnemius was observed by hematoxylin-eosin staining.The ultrastructure of gastrocnemius muscle was observed by transmission electron microscopy.Apoptosis of gastrocnemius cells was observed by TUNEL staining.The expressions of interleukin-1β,interleukin-6 and interleukin-10 in gastrocnemius muscle were detected by ELISA.The protein expressions of BAX,BCL-2 and Caspase3 in gastrocnemius muscle were detected by western blot.The mRNA expression of BAX and BCL-2 was detected by RT-PCR.RESULTS AND CONCLUSION:(1)Hematoxylin-eosin staining results showed that compared with the model group,inflammatory cells decreased in number,myocyte amount increased,and muscle tissue gradually healed in the Lijin group at each observation point.(2)The results of transmission electron microscopy showed that compared with the model group,the arrangement of muscle fibers at each observation point in the Lijin group was gradually orderly,mitochondria were gradually complete,Z-line arrangement was gradually regular,and free ribosomes were gathered.(3)TUNEL staining results showed that compared with the model group,apoptosis rate in the Lijin group was gradually decreased at all observation points(P<0.05).(4)ELISA results showed that compared with the model group,the expression of interleukin-1β and interleukin-6 in the Lijin group continued to decrease(P<0.05),while the expression of interleukin-10 increased on the 7th day after modeling,and then showed a downward trend(P<0.05).(5)Western blot results showed that compared with the model group,the expression of BCL-2 protein/BAX protein in the Lijin group was significantly increased at each observational point(P<0.05).The protein expression of Caspase3 decreased significantly(P<0.001),and was gradually similar to that of the blank group.(6)RT-PCR results showed that compared with the model group,the mRNA expression level of BCL-2/BAX in the Lijin group was significantly higher at each observational point(P<0.05).To conclude,Lijin manipulation can inhibit inflammation,reduce apoptosis,and promote the repair of injured skeletal muscle.

BACKGROUND:Remyelination in the central nervous system is a basic repair process triggered by demyelinating events,mainly through the proliferation,migration,and differentiation of oligodendrocyte precursor cells into oligodendrocytes.The process of remyelination is affected by many factors such as astrocytes,myelin debris,microglia,macrophages,endothelial cells,pericytes,T cells,and age.OBJECTIVE:Astrocytes play an important role in regulating synaptic activity,nutritional support,and tissue repair in the central nervous system.This review aims to provide potential therapeutic targets for demyelinating diseases of central nervous system by reviewing the role of astrocytes in remyelination.METHODS:A search was conducted on relevant literature collected from CNKI,PubMed,and Web of Science from 2014 tO 2024.The search terms were"astrocytes,oligodendrocyte precursor cells,remyelination"in both Chinese and English.Finally,66 articles were included after screening and summarized.RESULTS AND CONCLUSION:(1)The treatment of demyelinating diseases,such as multiple sclerosis,is limited to disease-modifying therapies,and there is no available method to overcome the failure of remyelination.Therefore,it is necessary to explore targets related to remyelination to promote myelin repair.(2)Remyelination is a process in which oligodendrocyte precursor cells proliferate,migrate,differentiate,and mature into oligodendrocytes,and the latter produce myelin to wrap axons to form myelin sheath.(3)Astrocytes regulate remyelination by phagocytosis of myelin debris,participating in inflammatory response,transforming into oligodendrocyte lineage cells,providing energy supply for oligodendrocyte lineage cells,releasing neurotrophic factors,and secreting extracellular matrix components.(4)The drugs screened in this paper use astrocytes and their derived factors as intervention targets to regulate the remyelination.Some drugs have satisfactory effects,but their effectiveness and safety still need more basic research and clinical trials to verify.(5)The mechanism of action of astrocytes in remyelination has not been fully elucidated,and the related molecular targets and signaling pathways can be further studied.

Wu Jutong's academic thoughts are inherited from Ye Tianshi and traced back to Zhongjing,and have a unique under-standing and play of the evil invading Shaoyang.Wu Jutong believes that Shaoyang has two major functional characteristics:ascending and pivoting.The principle of treating evil invading Shaoyang is to follow the ascending nature of Shaoyang and restore the pivoting function of Shaoyang.For Shaoyang fire-heat syndrome,follow Shaoyang to ascend and disperse to penetrate heat,and often use light,scavenging and dispersing products;for Shaoyang phlegm-fluid syndrome,creat Xiangfu Xuanfuhua Decoction to benefit the liver and gallbladder and resolve phlegm-fluid;for malaria invading Shaoyang,turn the pivot to stop malaria,and use Xiaochaihu Decoction and Qinghao Biejia Decoction for the treatment;for yin evil invading Shaoyang and Jueyin,Xiao Chaihu Decoction and Bupleurum chinense DC.are often used to turn the pivot of Shaoyang and warm and unchoke Jueyin.The contraindications of Xiao Chaihu Decoction are al-so summarized.

BACKGROUND:Lijin manipulation can reduce fibrosis scar hyperplasia and promote skeletal muscle repair.However,improper activation of the Wnt/β-catenin signaling pathway can aggravate the fibrosis of injured skeletal muscle and adversely affect the repair process of skeletal muscle.To study the regulatory effect of Lijin manipulation on the Wnt/β-catenin signaling pathway is conducive to elucidate the related mechanisms of Lijin manipulation in reducing fibrosis scar hyperplasia and promoting skeletal muscle injury repair.OBJECTIVE:To explore the mechanism of Lijin manipulation in promoting the repair of skeletal muscle injury in rabbits.METHODS:Forty-five healthy adult Japanese white rabbits were randomly divided into blank group,model group and Lijin group with 15 rabbits in each group.Gastrocnemius muscle percussion modeling was performed in both model group and Lijin group.Lijin manipulation was performed in the Lijin group on the 3rd day after modeling,once a day,15 minutes once.Five animals in each group were selected and killed on the 7th,14th and 21st days after modeling.The general morphological structure of gastrocnemius was observed by hematoxylin-eosin staining and the content of collagen fiber was observed by Masson staining.Western blot was used to detect the protein expression of Wnt3a,β-catenin,GSK3β,p-GSK3β,TCF,type I collagen and type III collagen in gastrocnemius muscle,and RT-PCR was used to detect the mRNA expression of Wnt3a,β-catenin and TCF.The expression of β-catenin was detected by immunofluorescence,and the expression of type I collagen and type III collagen was detected by immunohistochemistry.RESULTS AND CONCLUSION:The results of hematoxylin-eosin staining and Masson staining showed that compared with the model group,inflammatory cell infiltration and collagen fiber amount decreased in the Lijin group(P<0.001),and muscle fibers gradually healed.Western blot results showed that compared with the model group,the protein expression levels of Wnt3a,β-catenin,TCF,type I collagen and type III collagen were significantly decreased in the Lijin group at all observation time points(P<0.05),while the ratio of P-GSK3β/GSK3β was significantly increased in the Lijin group at all observation time points compared with the model group(P<0.05).RT-PCR results showed that compared with the model group,the mRNA expression levels of Wnt3a,β-catenin and TCF were significantly decreased in the Lijin group at all observation time points(P<0.001).Immunofluorescence results showed that compared with the model group,the fluorescence intensity of β-catenin expression in the Lijin group was significantly decreased at each observation time point and gradually became similar to that in the blank group(P<0.001).Immunohistochemical results showed that the expression levels of type I collagen and type III collagen in the Lijin group were significantly lower than those in the model group(P<0.01).To conclude,Lijin manipulation could inhibit the abnormal activation of the Wnt/β-catenin signaling pathway,reduce fibrotic scar hyperplasia,and promote the repair of injured skeletal muscle.

BACKGROUND:Excessive apoptosis in skeletal muscle cells will destroy the dynamic balance of the number of myocytes,leading to pathological injury of skeletal muscle.Lijin manipulation is effective in treating skeletal muscle injury,but whether it can inhibit apoptosis and promote the repair of skeletal muscle injury is unknown.OBJECTIVE:To explore the mechanism by which Lijin manipulation reduces inflammation and apoptosis during the repair of skeletal muscle injury in rabbits.METHODS:Forty-five healthy adult Japanese white rabbits were randomly divided into blank group,model group and Lijin group(n=15 per group).No intervention was performed in the blank group.Gastrocnemius muscle percussion molding was performed in both the model group and Lijin group.After modeling,the model group was not treated,while the Lijin manipulation(Stroking,kneading,and rubbing)was performed in the Lijin group on the 3rd day,once a day,15 min/time.Sampling in each group was performed on the 7th,14th and 21st days after modeling.The general morphological structure of gastrocnemius was observed by hematoxylin-eosin staining.The ultrastructure of gastrocnemius muscle was observed by transmission electron microscopy.Apoptosis of gastrocnemius cells was observed by TUNEL staining.The expressions of interleukin-1β,interleukin-6 and interleukin-10 in gastrocnemius muscle were detected by ELISA.The protein expressions of BAX,BCL-2 and Caspase3 in gastrocnemius muscle were detected by western blot.The mRNA expression of BAX and BCL-2 was detected by RT-PCR.RESULTS AND CONCLUSION:(1)Hematoxylin-eosin staining results showed that compared with the model group,inflammatory cells decreased in number,myocyte amount increased,and muscle tissue gradually healed in the Lijin group at each observation point.(2)The results of transmission electron microscopy showed that compared with the model group,the arrangement of muscle fibers at each observation point in the Lijin group was gradually orderly,mitochondria were gradually complete,Z-line arrangement was gradually regular,and free ribosomes were gathered.(3)TUNEL staining results showed that compared with the model group,apoptosis rate in the Lijin group was gradually decreased at all observation points(P<0.05).(4)ELISA results showed that compared with the model group,the expression of interleukin-1β and interleukin-6 in the Lijin group continued to decrease(P<0.05),while the expression of interleukin-10 increased on the 7th day after modeling,and then showed a downward trend(P<0.05).(5)Western blot results showed that compared with the model group,the expression of BCL-2 protein/BAX protein in the Lijin group was significantly increased at each observational point(P<0.05).The protein expression of Caspase3 decreased significantly(P<0.001),and was gradually similar to that of the blank group.(6)RT-PCR results showed that compared with the model group,the mRNA expression level of BCL-2/BAX in the Lijin group was significantly higher at each observational point(P<0.05).To conclude,Lijin manipulation can inhibit inflammation,reduce apoptosis,and promote the repair of injured skeletal muscle.

Objective To establish and evaluate a mouse model of chronic obstructive pulmonary disease(COPD)induced by cigarette smoke(CS).Methods Forty BALB/c mice were divided randomly into a control group and a CS group.Mice in the CS group were subjected to passive smoking for 20 weeks and a COPD model was established.Morphological changes in the organs and lung,heart,liver,and kidney fibrosis were observed by hematoxylin-eosin(HE)and Masson staining.Lung,cardiac,and brain cognitive function were evaluated by pulmonary function testing,small-animal ultrasound,and Morris water maze trials.Tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-1β levels in lung and brain tissues were detected by ELISA.Liver and renal functions were measured by biochemical method.Results The alveolar septum was narrowed or broken in mice in the CS group,and the adjacent alveolar cavity was enlarged and fused,consistent with the pathological changes of COPD.Neuronal degeneration and necrosis were observed in the hippocampus,but there were no significant morphological changes in other organs.Masson staining showed no obvious fibrosis in the lung,heart,liver,or kidney in CS-group mice.The result of pulmonary function tests showed that the forced expiratory volume in 0.1 second/forced vital capacity(FEV 0.1/FVC)and dynamic compliance were significantly decreased in the CS group compared with the control group,while airway resistance was obviously increased.Cognitive impairment in mice in the CS group was confirmed in the Morris water maze trial.TNF-α,IL-6,and IL-1β levels in lung and brain tissues were higher in the CS group compared with the control group.There were no significant differences in cardiac,liver,and renal functions between the groups.Conclusions A mouse model of COPD can be established by CS exposure for 20 weeks.Lung histomorphology,lung function,brain cognitive function,and levels of inflammatory factors can be used as indicators to evaluate the success of the model.