Objective To investigate whether miR-15b-5p can alleviate airway inflammation in asthma by negatively regulating ubiquitin specific peptidase 9X (USP9X) to down-regulate the expression of phosphatidylinositol 4, 5-diphosphate 3-kinase catalytic subunit α/AKT serine/threonine kinase 1 (PIK3CA/AKT1) pathway. Methods USP9X was predicted to be a direct target of miR-15b-5p by using an online database (miRWalk), and the luciferase reporter gene assay was performed to verify it. Co-immunoprecipitation (CO-IP) was used to verify the direct binding between USP9X and PIK3CA and the role of USP9X and its small molecule inhibitor WP1130 in the deubiquitination of PIK3CA. C57 mice were randomly divided into Control group, OVA group, OVA combined with NC group and miR-15b-5p agomir group, with 10 mice in each group. BEAS-2B cells were induced with interleukin 13 (IL-13) and treated with miR-15b-5p mimic. HE, Masson, PAS, immunohistochemistry, immunofluorescence staining, flow cytometry, Western blot and quantitative real-time PCR(qRT-PCR) were performed. Results It was found that the administration of miR-15b-5p agomir and mimic could reduce peribronchial inflammatory cells and improve airway inflammation, and miR-15b-5p could target negative regulation of USP9X. USP9X could directly bind to PIK3CA and regulate PIK3CA level in a proteasome-dependent manner, and USP9X could deubiquitinate K29-linked PIK3CA protein. Down-regulation of USP9X could increase PIK3CA ubiquitination level. WP1130, a small molecule inhibitor of USP9X, has the same effect as knockdown of USP9X, both of which could increase the ubiquitination level of PIK3CA and reduce the protein level of PIK3CA. Conclusion The miR-15b-5p/USP9X/PIK3CA/AKT1 signaling pathway may provide potential therapeutic targets for asthma.
Animals ; MicroRNAs/metabolism* ; Asthma/pathology* ; Class I Phosphatidylinositol 3-Kinases/genetics* ; Ubiquitin Thiolesterase/metabolism* ; Proto-Oncogene Proteins c-akt/genetics* ; Mice ; Signal Transduction ; Mice, Inbred C57BL ; Humans ; Inflammation/genetics* ; Cell Line ; Female ; Male

Ischemic stroke (IS) is a globally life-threatening disease. Presently, few therapeutic medicines are available for treating IS, and rt-PA is the only drug approved by the US Food and Drug Administration (FDA) in the US. In fact, many agents showing excellent neuroprotection but no blood flow-improving activity in animals have not achieved ideal clinical efficacy, while thrombolytic drugs only improving blood flow without neuroprotection have limited their wider application. To address these challenges and meet the huge unmet clinical need, we have designed and identified a novel compound AAPB with dual effects of neuroprotection and cerebral blood flow improvement. AAPB significantly reduced cerebral infarction and neural function deficit in tMCAO rats, pMCAO rats, and IS rhesus monkeys, as well as displayed exceptional safety profiles and excellent pharmacokinetic properties in rats and dogs. AAPB has now entered phase I of clinical trials fighting IS in China.

Objective To investigate the changes and clinical significance of pyroptosis and inflammation in children with acute lymphoblastic leukemia after chemotherapy.Methods A retrospective analysis was con-ducted on the clinical data of 98 children with acute lymphoblastic leukemia who received chemotherapy in the pediatrics and hematology and oncology departments of the hospital from May 2023 to August 2024.Accord-ing to the results of blood culture,the selected children were divided into the Gram-positive bacteria group,the Gram-negative bacteria group,the fungal group and the non-bloodstream infection group,and drug sensitivity tests were conducted.After chemotherapy,the children were divided into the granulocytosis group and the non-granulocytosis group according to the granulocyte level.The relevant indicators were detected and com-pared by methods such as blood routine,flow microsphere array technology,enzyme-linked immunosorbent assay(ELISA),and Western blot.Results After chemotherapy,the pyroptosis related indicators caspase-1,caspase-4,caspase-5,caspase-11,interleukin(IL)-1 β,IL-18,the proportion of pyroptosis cells and the relative expression level of GSDMD protein in children of each infection type were significantly increased compared with those before chemotherapy(P＜0.05).After chemotherapy,the levels of IL-4,IL-6,IL-10 and interfer-on-γ(IFN-γ)in the granulocytosis group were significantly higher than those in the non-granulocytosis group(P＜0.05),and the granulocyte level was negatively correlated with the levels of IL-4,IL-6,IL-10 and IFN-γ(P＜0.05).There were statistically significant differences in the levels of IL-4,IL-6,IL-10 and IFN-γ in dif-ferent infection states after chemotherapy(P＜0.05).Conclusion The number of granulocytes and the levels of serum cytokines can serve as potential indicators of infection in children with leukemia.The regulation of pyroptosis may provide new strategies for the treatment of childhood leukemia.

BACKGROUND:Stem cell transplantation has a promising therapeutic prospect in the treatment of myocardial infarction,but the efficacy of stem cell transplantation is limited by the low homing efficiency of transplanted cells to the heart and the low retention rate and survival rate in the heart.Exercise therapy is an important integral component of cardiac rehabilitation for patients with myocardial infarction.However,the role of exercise in stem cell therapy for myocardial infarction has not yet been clarified.OBJECTIVE:To investigate the effect of exercise(including acute exercise and chronic exercise)on bone marrow mesenchymal stem cell transplantation in rats with myocardial infarction.METHODS:Eighty female SD rats were randomly divided into sham operation group,model group,transplantation group or combination group with random number table method(n=20).Myocardial infarction model of rats in model group,transplantation group,or combination group was made by coronary artery ligation.24 hours after the model was made,the combination group underwent aerobic exercise for 8 weeks(chronic exercise,30 min/d,5 days per week),and within 5 minutes after the first exercise(acute exercise).SD rat bone marrow mesenchymal stem cells labeled with green fluorescent protein were injected into the tail vein of the transplantation group and the combination group.A part of animals from each group were taken 24 hours after the first exercise.The survival rate of stem cells transplanted into rat myocardium,sex-determining region of Y,protein expression of homing factors,oxidative stress,and inflammatory response parameters were measured.After 72 hours of the last exercise,the remaining rats were taken to detect cardiac structure and function,myocardial histological changes,and the number of Ki67+cells.RESULTS AND CONCLUSION:(1)After acute exercise:Compared with sham operation group,myocardial reactive oxygen species level,malondialdehyde content,tumor necrosis factor-α,and interleukin-1β protein expression increased(P<0.05),and superoxide dismutase activity decreased(P<0.05)in model group.Compared with model group,reactive oxygen species,malondialdehyde content,tumor necrosis factor-α,and interleukin-1β protein expression reduced(P<0.05),superoxidation dismutase activity,stromal cell-derived factor 1α,and CXC chemokine receptor 4 protein expression increased(P<0.05)in transplantation and combination groups.Compared with the transplantation group,reactive oxygen species,malondialdehyde content,tumor necrosis factor-α,and interleukin-1β protein expression decreased(P<0.05),stem cell survival rate,sex-determining region of Y mRNA expression,superoxide dismutase activity,stromal cell-derived factor 1α,and CXC chemokine receptor 4 protein expression increased(P<0.05)in combination group.(2)After chronic exercise:Compared with sham operation group,cardiomyocyte cross-sectional area and collagen content increased(P<0.05),left ventricular ejection fraction and left ventricular short-axis shortening rate decreased(P<0.05)in model group.Compared with model group,cardiomyocyte cross-sectional area and collagen content decreased(P<0.05),Ki67+cells increased(P<0.05)in transplantation group.Compared with transplantation group,collagen content decreased(P<0.05),cardiomyocyte cross-sectional area,left ventricular ejection fraction,left ventricular short-axis shortening rate,and Ki67+cells increased(P<0.05)in the combination group.(3)Acute exercise improves the survival rate of exogenous stem cells by promoting stem cell homing and improving myocardial microenvironment,while chronic exercise can stimulate cardiomyocyte proliferation,inhibit cardiac remodeling,and enhance cardiac function after stem cell transplantation.Therefore,exercise can help to optimize the efficacy of stem cell transplantation after myocardial infarction in rats.

Objective To explore the mechanism of Glaucocalyxin A(GLA)in inhibiting ovalbumin(OVA)-induced airway remodeling in asthmatic mice through the topoisomerase Ⅱ α(TOP2A)/cyclin-dependent kinase 1(CDK1)signaling pathway.Methods Forty Balb/c mice were randomly divided into 5 groups:the control group,the model group,the low-dose GLA group,the high-dose GLA group and the Dexamethasone group,with 8 mice in each group.The effect of GLA on airway remodeling was examined by immunohistochemical staining,ELISA and other methods,and bioinformatics methods were used to predict new targets of GLA.The action targets of TOP2A were screened using the STRING database,and the interaction relationship between the two was verified by co-immunoprecipitation.In vitro,GLA and siRNA were used to interfere with interleukin-4(IL-4)-stimulated human airway epithelial cells BEAS-2B.The expressions of TOP2A,epidermal growth factor receptor(EGFR),Integrin β1,focal adhesion kinase(FAK),β-catenin,CDK1 and DRP1 were detected by Western Blot.Results GLA intervention could significantly reduce OVA-induced asthma airway remodeling,airway smooth muscle thickening,collagen deposition around the airway,the number of eosinophils in alveolar lavage fluid,the expression of pro-inflammatory cytokines such as IL-4,and the level of serum IgE.The new target of GLA screened out was TOP2A,which was highly expressed in the lung tissue of the asthma airway remodeling model.GLA intervention could down-regulate its expression.In vitro,intervention with GLA and si-TOP2A could significantly down-regulate the expressions of IL-4-induced TOP2A,EGFR,Integrin β1,FAK and β-catenin.Further studies have found that TOP2A had an interaction relationship with CDK1.si-TOP2A could downregulate the expression of CDK1,and knockdown of CDK1 could significantly down-regulate the expression of phosphorylated DRP1.Conclusion GLA may alleviate asthma airway remodeling by targeting the TOP2A/CDK1 signaling pathway,providing experimental evidence for the clinical diagnosis and treatment of asthma airway remodeling in asthma.

Objective To establish the therapeutic effect prediction model of platelet transfusion in hematological patients,and receiver operating characteristic(ROC)curve and clinical cases are used to evaluate the clinical application value of the predic-tion model.Methods A total of 485 patients with hematological diseases who received platelet transfusion therapy in Kailuan General Hospital from January 2020 to December 2023 were selected,corrected count increment(CCI)was used to divide the patients into platelet transfusion effective group(n=340)and transfusion ineffective group(n=145).Multivariate Logistic regres-sion analysis was used to establish the prediction model of platelet infusion efficacy,and ROC curve was used to evaluate the application effect of the forcasting model.109 clinical cases were used to verify the practical application effect of the model,and the sensitivity,specificity and accuracy were calculated.Results Among 485 patients with hematological diseases,the incidence of ineffective platelet transfusion was 29.90%(145/485).Compated with the effective group,the ineffective group had more previous platelet transfusions was higher,and the difference was statistically significant(t=-4.435,P<0.05).In the ineffective group,there were more cases of hyperplenism,aplastic anemia and lymphoma,higher infection rate and higher positive rate of platelet antibody,and the differences were statistically significant(χ2=6.301～37.522,all P<0.05).Multivariate Logistic regres-sion analysis found that previous platelet infusion times,infection,leukemia,aplastic anemia and platelet antibodies were risk factors for ineffective platelet transfusion in patients with hematological diseases(Wald χ2=5.224～21.548,all P<0.05).Based on these risk factors,platelet infusion effect prediction models 1 and 2 were constructed.ROC curve was used to evaluate the application effect of the prediction model.The area under the curve(AUC),cut-offpoint,sensitivity and specificity of model 1 were 0.884,0.042,82.35%,88.89%.The AUC,cut-offpoint,corresponding sensitivity and specificity of prediction model 2 were 0.910,59.784,81.18%,94.44%,respectively.The Z values of model 1 and model 2 were 12.159 and 13.151,respectively.The prediction effect of model 2 was better than that of model 1.The actual application results showed that the sensitivity,specificity and accuracy of prediction model 1,2 were 85.71%,92.05%,90.89%and 90.48%,93.18%,92.66%,respectively.Conclusion The ineffective rate of platelet transfusion in hematological patients is relatively high.The prediction models 1 and 2 for platelet transfusion effectiveness have good results in predicting ineffective platelet transfusion,and prediction model 2 is better than pre-diction model 1,which can provide reliable basis for hematological patients on accurate platelet transfusion.

Objective To investigate the value of serum α-hydroxybutyric dehydrogenase(α-HBDH),cysteine-rich protein 61(CYR61)and gasdermin D(GSDMD)level testing in patients with sepsis-combined cardiomyopathy for clinical diagnosis and prognostic assessment.Methods A total of 244 sepsis patients who underwent consultation and treatment in Baotou Central Hospital from May 2020 to December 2023 were selected as the study subjects,and were separated into a study group(combined cardiomyopathy,n=106)and a control group(uncombined cardiomyopathy,n=138)according to whether they were combined cardiomyopathy or not.The levels of α-HBDH,CYR61 and GSDMD were measured by enzyme linked immunosorbent assay(ELISA)method.Pearson and Spearman methods were used to analyze the correlation of α-HBDH,CYR61,and GSDMD with systolic and diastolic blood pressure,left ventricular ejection fraction(LVEF)and acute physiology and chronic health evaluationⅡ(APACHE II)score.Multifactorial Logistic regression was used to analyze the factors affecting sepsis-combined cardiomyopathy.Receiver operator characteristic(ROC)curves were used to assess the diagnostic value of α-HBDH,CYR61 and GSDMD for sepsis-combined cardiomyopathy and their validity for prognostic prediction.Results Serum α-HBDH(278.35±18.89ng/ml vs 253.47±12.75ng/ml),CYR61(18.23±4.14mg/L vs 14.48±2.67mg/L)and GSDMD(12.39±3.28mg/L vs 9.46±2.17mg/L)levels were higher in the study group compared to the control group,and the differences were statistically significant(t=12.261,8.572,8.377,all P<0.05).The levels of α-HBDH(291.93±19.22ng/ml),CYR61(20.33±3.43mg/L)and GSDMD(14.01±3.09mg/L)were higher in the death patients compared to the survived patients(268.71±13.09ng/ml,16.74±2.88mg/L,11.24±2.55mg/L),and the differences were statistically significant(t=7.402,5.839,5.044,all P<0.05).Correlation analysis showed that α-HBDH,CYR61,and GSDMD were negatively correlated with systolic blood pressure,diastolic blood pressure and LVEF(r=-0.631～-0.422,all P<0.05),α-HBDH,CYR61,GSDMD were negatively correlated with APACHE II score(r=0.531,0.507,0.611,all P<0.05).Multifactorial Logistic regression analysis showed that systolic blood pressure,diastolic blood pressure,and LVEF were protective factors affecting sepsis-combined cardiomyopathy(Wald χ2=6.823,7.986,10.875,all P<0.05),and α-HBDH,CYR61,and GSDMD were risk factors affecting sepsis-combined cardiomyopathy(Wald χ2=9.376,6.849,7.435,all P<0.05).From the ROC curve analysis,it was known that the combined application of α-HBDH,CYR61,and GSDMD was more effective in the diagnosis of sepsis-combined cardiomyopathy(Z=2.369,2.454,2.573),the combined application of α-HBDH,CYR61,and GSDMD were superior for prognostic prediction in sepsis-combined cardiomyopathy(Z=2.352,2.468,2.581),and the differences were statistically significant(all P<0.05).Conclusion Serum α-HBDH,CYR61 and GSDMD levels are increased in patients with sepsis-combined cardiomyopathy,and they are correlated with prognosis.The combination of these three tests has a higher diagnostic value and prognostic value in sepsis combined cardiomyopathy.

Objective To explore the mechanism of Glaucocalyxin A(GLA)in inhibiting ovalbumin(OVA)-induced airway remodeling in asthmatic mice through the topoisomerase Ⅱ α(TOP2A)/cyclin-dependent kinase 1(CDK1)signaling pathway.Methods Forty Balb/c mice were randomly divided into 5 groups:the control group,the model group,the low-dose GLA group,the high-dose GLA group and the Dexamethasone group,with 8 mice in each group.The effect of GLA on airway remodeling was examined by immunohistochemical staining,ELISA and other methods,and bioinformatics methods were used to predict new targets of GLA.The action targets of TOP2A were screened using the STRING database,and the interaction relationship between the two was verified by co-immunoprecipitation.In vitro,GLA and siRNA were used to interfere with interleukin-4(IL-4)-stimulated human airway epithelial cells BEAS-2B.The expressions of TOP2A,epidermal growth factor receptor(EGFR),Integrin β1,focal adhesion kinase(FAK),β-catenin,CDK1 and DRP1 were detected by Western Blot.Results GLA intervention could significantly reduce OVA-induced asthma airway remodeling,airway smooth muscle thickening,collagen deposition around the airway,the number of eosinophils in alveolar lavage fluid,the expression of pro-inflammatory cytokines such as IL-4,and the level of serum IgE.The new target of GLA screened out was TOP2A,which was highly expressed in the lung tissue of the asthma airway remodeling model.GLA intervention could down-regulate its expression.In vitro,intervention with GLA and si-TOP2A could significantly down-regulate the expressions of IL-4-induced TOP2A,EGFR,Integrin β1,FAK and β-catenin.Further studies have found that TOP2A had an interaction relationship with CDK1.si-TOP2A could downregulate the expression of CDK1,and knockdown of CDK1 could significantly down-regulate the expression of phosphorylated DRP1.Conclusion GLA may alleviate asthma airway remodeling by targeting the TOP2A/CDK1 signaling pathway,providing experimental evidence for the clinical diagnosis and treatment of asthma airway remodeling in asthma.

Objective To establish the therapeutic effect prediction model of platelet transfusion in hematological patients,and receiver operating characteristic(ROC)curve and clinical cases are used to evaluate the clinical application value of the predic-tion model.Methods A total of 485 patients with hematological diseases who received platelet transfusion therapy in Kailuan General Hospital from January 2020 to December 2023 were selected,corrected count increment(CCI)was used to divide the patients into platelet transfusion effective group(n=340)and transfusion ineffective group(n=145).Multivariate Logistic regres-sion analysis was used to establish the prediction model of platelet infusion efficacy,and ROC curve was used to evaluate the application effect of the forcasting model.109 clinical cases were used to verify the practical application effect of the model,and the sensitivity,specificity and accuracy were calculated.Results Among 485 patients with hematological diseases,the incidence of ineffective platelet transfusion was 29.90%(145/485).Compated with the effective group,the ineffective group had more previous platelet transfusions was higher,and the difference was statistically significant(t=-4.435,P<0.05).In the ineffective group,there were more cases of hyperplenism,aplastic anemia and lymphoma,higher infection rate and higher positive rate of platelet antibody,and the differences were statistically significant(χ2=6.301～37.522,all P<0.05).Multivariate Logistic regres-sion analysis found that previous platelet infusion times,infection,leukemia,aplastic anemia and platelet antibodies were risk factors for ineffective platelet transfusion in patients with hematological diseases(Wald χ2=5.224～21.548,all P<0.05).Based on these risk factors,platelet infusion effect prediction models 1 and 2 were constructed.ROC curve was used to evaluate the application effect of the prediction model.The area under the curve(AUC),cut-offpoint,sensitivity and specificity of model 1 were 0.884,0.042,82.35%,88.89%.The AUC,cut-offpoint,corresponding sensitivity and specificity of prediction model 2 were 0.910,59.784,81.18%,94.44%,respectively.The Z values of model 1 and model 2 were 12.159 and 13.151,respectively.The prediction effect of model 2 was better than that of model 1.The actual application results showed that the sensitivity,specificity and accuracy of prediction model 1,2 were 85.71%,92.05%,90.89%and 90.48%,93.18%,92.66%,respectively.Conclusion The ineffective rate of platelet transfusion in hematological patients is relatively high.The prediction models 1 and 2 for platelet transfusion effectiveness have good results in predicting ineffective platelet transfusion,and prediction model 2 is better than pre-diction model 1,which can provide reliable basis for hematological patients on accurate platelet transfusion.

Objective To investigate the value of serum α-hydroxybutyric dehydrogenase(α-HBDH),cysteine-rich protein 61(CYR61)and gasdermin D(GSDMD)level testing in patients with sepsis-combined cardiomyopathy for clinical diagnosis and prognostic assessment.Methods A total of 244 sepsis patients who underwent consultation and treatment in Baotou Central Hospital from May 2020 to December 2023 were selected as the study subjects,and were separated into a study group(combined cardiomyopathy,n=106)and a control group(uncombined cardiomyopathy,n=138)according to whether they were combined cardiomyopathy or not.The levels of α-HBDH,CYR61 and GSDMD were measured by enzyme linked immunosorbent assay(ELISA)method.Pearson and Spearman methods were used to analyze the correlation of α-HBDH,CYR61,and GSDMD with systolic and diastolic blood pressure,left ventricular ejection fraction(LVEF)and acute physiology and chronic health evaluationⅡ(APACHE II)score.Multifactorial Logistic regression was used to analyze the factors affecting sepsis-combined cardiomyopathy.Receiver operator characteristic(ROC)curves were used to assess the diagnostic value of α-HBDH,CYR61 and GSDMD for sepsis-combined cardiomyopathy and their validity for prognostic prediction.Results Serum α-HBDH(278.35±18.89ng/ml vs 253.47±12.75ng/ml),CYR61(18.23±4.14mg/L vs 14.48±2.67mg/L)and GSDMD(12.39±3.28mg/L vs 9.46±2.17mg/L)levels were higher in the study group compared to the control group,and the differences were statistically significant(t=12.261,8.572,8.377,all P<0.05).The levels of α-HBDH(291.93±19.22ng/ml),CYR61(20.33±3.43mg/L)and GSDMD(14.01±3.09mg/L)were higher in the death patients compared to the survived patients(268.71±13.09ng/ml,16.74±2.88mg/L,11.24±2.55mg/L),and the differences were statistically significant(t=7.402,5.839,5.044,all P<0.05).Correlation analysis showed that α-HBDH,CYR61,and GSDMD were negatively correlated with systolic blood pressure,diastolic blood pressure and LVEF(r=-0.631～-0.422,all P<0.05),α-HBDH,CYR61,GSDMD were negatively correlated with APACHE II score(r=0.531,0.507,0.611,all P<0.05).Multifactorial Logistic regression analysis showed that systolic blood pressure,diastolic blood pressure,and LVEF were protective factors affecting sepsis-combined cardiomyopathy(Wald χ2=6.823,7.986,10.875,all P<0.05),and α-HBDH,CYR61,and GSDMD were risk factors affecting sepsis-combined cardiomyopathy(Wald χ2=9.376,6.849,7.435,all P<0.05).From the ROC curve analysis,it was known that the combined application of α-HBDH,CYR61,and GSDMD was more effective in the diagnosis of sepsis-combined cardiomyopathy(Z=2.369,2.454,2.573),the combined application of α-HBDH,CYR61,and GSDMD were superior for prognostic prediction in sepsis-combined cardiomyopathy(Z=2.352,2.468,2.581),and the differences were statistically significant(all P<0.05).Conclusion Serum α-HBDH,CYR61 and GSDMD levels are increased in patients with sepsis-combined cardiomyopathy,and they are correlated with prognosis.The combination of these three tests has a higher diagnostic value and prognostic value in sepsis combined cardiomyopathy.