Objective To investigate the potential causal associations between 3 common subjective sleep problems,daytime sleepiness,insomnia,and sleep apnea,and epigenetic age acceleration.Methods A two-sample Mendelian randomization(MR)study was conducted the publicly available genome-wide association study(GWAS)data to assess the causal effects of self-reported sleep problems on 4 indicators of epigenetic age acceleration,including intrinsic epigenetic age acceleration(IEAA),GrimAge acceleration,HannumAge acceleration,and PhenoAge acceleration.The primary analytical method was inverse-variance weighting(IVW),supplemented by sensitivity analyses using MR-Egger regression and the weighted median(WM)method to examine the robustness of the findings.Results IVW showed that daytime sleepiness was significantly positively associated with IEAA(β=1.83,95%CI:0.17～2.49,P=0.031)and HannumAge acceleration(β=1.81,95%CI:0.21～2.42,P=0.027),suggesting a potential accelerating effect on epigenetic aging.Insomnia also showed significant positive associations with IEAA(β=1.57,95%CI:0.40～2.73,P=0.009)and GrimAge acceleration(β=1.37,95%CI:0.18～2.56,P=0.024).No significant causal relationship was observed between sleep apnea and any indicator of epigenetic age acceleration(P>0.05).Conclusion Daytime sleepiness and insomnia may accelerate epigenetic aging,indicating their potential biological role in the aging process.The impact of sleep apnea remains unclear and warrants further investigation.

Objective To investigate the anti-hepatocellular carcinoma mechanism of Chloranthus fortunei(A.Gray)Solms-Laub.via network pharmacology,molecular docking techniques and in vitro experiments.Methods Chemical composition of Chloranthus fortunei(A.Gray)Solms-Laub.was searched by literature.Swiss Target Prediction was used to find corresponding targets.STRING was used to construct protein-protein interactions network(PPI).DAVID was used to enrich GO analysis and KEGG pathway.AutoDock Vina 1.1.2 and Pymol visualisation was used for docking and validation.Results Chloranthus fortunei(A.Gray)Solms-Laub.had 61 active components,685 targets,and 279 intersections with disease targets.The PPI showed that the main active components were Luteolin,Chloranthalactone C,Shizukanolide H,Esculetin,7-Hydroxycoumarin.The key targets were GAPDH,VEGFA,STAT3,JUN,HSP90AA1,AKT1,CTNNB1,CASP3,and ALB.Biological process(BP)involved protein phosphorylation,signal transduction,regulation of RNA polymerase II promoter transcription,cell proliferation,apoptosis.Cellular component(CC)involved cytoplasm,nucleus,cell membrane,cellular exosome.Molecular function(MF)involves protein binding,ATPase,threonine kinase,protein kinase activity.KEGG involved cancer pathway,metabolic pathway,PI3K-Akt signalling pathway,cancer proteoglycans,lipids and atherosclerosis,cytomegalovirus infection,microRNAs in cancer,human T-cell leukaemia virus type 1,Ras signalling pathway,MAPK signalling pathway.Molecular docking showed that silverweed lactone H had a strong affinity for each of the other target proteins,indicating that this component plays a key role.The results of RT-qPCR assay and WB assay showed that there were significant differences in gene and protein expression levels before and after drug administration.Conclusion The Chinese medicine in Chloranthus fortunei(A.Gray)Solms-Laub.can treat hepatocellular carcinoma through the MAPK pathway,and the main active ingredients have good docking effects with the core target proteins of the disease.

Objective To investigate the value of whole lung CT radiomics combined with clinical and conventional CT features for differentiating non-tuberculous mycobacterial pulmonary disease(NTM-PD)and pulmonary tuberculosis(PTB).Methods Fifty-three NTM-PD(NTM-PD group)and 111 PTB(PTB group)patients diagnosed by mycobacteria culture were retrospectively collected.The patients were divided into training set(n=115,including 37 cases of NTM-PD and 78 cases of PTB)and test set(n=49,including 16 cases of NTM-PD and 33 cases of PTB)at the ratio of 7∶3.Patients'clinical and pulmonary CT manifestations of lesions were analyzed using univariate and multivariate logistic regression,and the independent impact factors for differentiating NTM-PD and PTB lesions were screened.The best radiomics features were extracted and screened based on whole lung CT.Based on independent impact factors,the best radiomics features and their combination,clinical-CT,radiomics and combined models were constructed with random forest,and the differential efficacy of each model was evaluated.Results Patients'age(OR=0.264),gender(OR=0.956),immunoglobulin G(OR=3.416),C reactive protein(OR=3.418)and bronchiectasis shown on CT(OR=0.285)were all independent impact factors for differentiating NTM-PD and PTB.Twelve best radiomics features were screened based on whole lung ROI.The AUC of combined model in training set and test set was 0.915 and 0.901,respectively,both higher than that of clinical model(AUC=0.832,0.801,Z=1.340,3.710,both P＜0.05)and radiomics model(AUC=0.877,0.821,Z=-2.520,-5.240,both P＜0.05).Conclusion Whole lung CT radiomics combined with clinical and conventional CT features could effectively distinguish NTM-PD and PTB.

Objective To explore the functional connectivity(FC)changes of hippocampus and amygdala in type 2 diabetes mellitus(T2DM)patients with erectile dysfunction(DMED),and the central pathological neural mechanisms underlying DMED.Methods 61 T2DM patients who visited Department of Endocrinology,Nanjing First Hospital,Nanjing Medical University from January 2020 to December 2021 were selected and divided into a simple T2DM group(n=30)and a combined DMED group(n=31).Another 47 healthy individuals were selected as control group(NC).The international erectile function scale(IIEF-5)was used to evaluate the erectile function.Resting-state functional magnetic resonance imaging(rs-fMRI)data were acquired from all participants.MRI data were preprocessed by the DPABI software package.Bilateral hippocampus and amygdala were selected as regions of interest(ROI)and the whole brain FC values were calculated.The FC values of brain regions between groups were tested by two-sample t-test with REST software package.Results Left hippocampus as ROI:compared with the NC group,FC values of the left superior temporal gyrus increased in the T2DM group,FC values of the left superior frontal gyrus,left inferior temporal gyrus,left posterior central gyrus and rectus gyrus decreased in the DMED group.Compared with the T2DM group,FC values of the left inferior parietal gyrus,left supramarginal gyrus,left middle occipital gyrus and right posterior central gyrus decreased in the DMED group.Right hippocampus as ROI:compared with the NC group,FC values of the right middle temporal gyrus and right rolandic operculum increased while FC values of the right calcarine fissure decreased in the T2DM group;FC values of bilateral anterior cingulate gyrus,right middle temporal gyrus and left rectus gyrus decreased in the DMED group.Compared with the T2DM group,FC values of the left middle frontal gyrus,left inferior parietal gyrus and right inferior temporal gyrus decreased in the DMED group.Left amygdala as ROI:compared with the NC group,FC values in the left parahippocampal gyrus,left fusiform gyrus and right insula increased in the T2DM group;FC value of the left middle temporal gyrus decreased in the DMED group.Compared with the T2DM group,FC values of the left middle frontal gyrus and left supramarginal gyrus decreased in the DMED group.Right amygdala as ROI:compared with the NC group,FC values of the left insula,right parahippocampal gyrus,right superior temporal gyrus and right supramarginal gyrus increased while FC values in the right caudate decreased in the T2DM group;FC values of the right middle frontal gyrus,left rectus gyrus and left middle occipital gyrus decreased in the DMED group.Compared with the T2DM group,FC values of the left middle frontal gyrus and left inferior parietal gyrus decreased in the DMED group.Conclusions DMED patients have abnormalities in the hippocampus,amygdala and other brain regions,especially the frontal lobe functional cortex,which may be related to changes in their brain function.

Objective To explore the relationship between the inhibitory effect of quercetin on AC16 cardiomyocyte proliferation and the Wnt/β-catenin signaling pathway,the relationship between quercetin(quercetin,QCT)and the proliferation of AC16 cardiomyocytes through in vitro was investigated.Methods AC16 cells were stimulated with different concentrations of QCT.The effects of QCT on AC16 cell proliferation were detected by inverted microscope photography,trypan blue counting,and CCK-8 assay.Western blot was used to detect the effects of QCT on the expression of phosphorylated β-catenin(p-β-catenin),c-Myc,β-catenin,low density lipoprotein receptor-related protein 5(LRP5),and LRP6.The effect of quercetin on cell proliferation was detected after overexpressing β-catenin ΔN,LRP5,and LRP6 genes,and after silencing LRP5 and LRP6 genes by trypan blue counting.Results Compared with the control group,QCT could decrease the number of AC16 cells and inhibit the proliferation rate,which was concentration-dependent.At the protein expression level,10 and 20 μmol/L QCT led to an significant upregulated modification of p-β-catenin protein(P<0.05)and significant downregulation of c-Myc,β-catenin,LRP5,and LRP6 protein expression(P<0.05)in AC16 cells.Therefore,10 μmol/L QCT was chosen as the intervention concentration.Overexpression of β-catenin ΔN,LRP5,and LRP6 genes in AC16 cells significantly rescued the cell proliferation inhibition caused by 10 μmol/L QCT compared to the drug-only group(P<0.05).Conversely,silencing LRP5 and LRP6 genes led to inhibition of AC16 cell proliferation,and the combination with 10 μmol/L QCT did not exacerbate the inhibition(P>0.05).Conclusion The quercetin could inhibit the Wnt/β-catenin signaling pathway via significant downregulation of LRP5/6,thereby attenuate cell proliferation of AC16 cardiomyocytes.

Objective To explore the functional connectivity(FC)changes of hippocampus and amygdala in type 2 diabetes mellitus(T2DM)patients with erectile dysfunction(DMED),and the central pathological neural mechanisms underlying DMED.Methods 61 T2DM patients who visited Department of Endocrinology,Nanjing First Hospital,Nanjing Medical University from January 2020 to December 2021 were selected and divided into a simple T2DM group(n=30)and a combined DMED group(n=31).Another 47 healthy individuals were selected as control group(NC).The international erectile function scale(IIEF-5)was used to evaluate the erectile function.Resting-state functional magnetic resonance imaging(rs-fMRI)data were acquired from all participants.MRI data were preprocessed by the DPABI software package.Bilateral hippocampus and amygdala were selected as regions of interest(ROI)and the whole brain FC values were calculated.The FC values of brain regions between groups were tested by two-sample t-test with REST software package.Results Left hippocampus as ROI:compared with the NC group,FC values of the left superior temporal gyrus increased in the T2DM group,FC values of the left superior frontal gyrus,left inferior temporal gyrus,left posterior central gyrus and rectus gyrus decreased in the DMED group.Compared with the T2DM group,FC values of the left inferior parietal gyrus,left supramarginal gyrus,left middle occipital gyrus and right posterior central gyrus decreased in the DMED group.Right hippocampus as ROI:compared with the NC group,FC values of the right middle temporal gyrus and right rolandic operculum increased while FC values of the right calcarine fissure decreased in the T2DM group;FC values of bilateral anterior cingulate gyrus,right middle temporal gyrus and left rectus gyrus decreased in the DMED group.Compared with the T2DM group,FC values of the left middle frontal gyrus,left inferior parietal gyrus and right inferior temporal gyrus decreased in the DMED group.Left amygdala as ROI:compared with the NC group,FC values in the left parahippocampal gyrus,left fusiform gyrus and right insula increased in the T2DM group;FC value of the left middle temporal gyrus decreased in the DMED group.Compared with the T2DM group,FC values of the left middle frontal gyrus and left supramarginal gyrus decreased in the DMED group.Right amygdala as ROI:compared with the NC group,FC values of the left insula,right parahippocampal gyrus,right superior temporal gyrus and right supramarginal gyrus increased while FC values in the right caudate decreased in the T2DM group;FC values of the right middle frontal gyrus,left rectus gyrus and left middle occipital gyrus decreased in the DMED group.Compared with the T2DM group,FC values of the left middle frontal gyrus and left inferior parietal gyrus decreased in the DMED group.Conclusions DMED patients have abnormalities in the hippocampus,amygdala and other brain regions,especially the frontal lobe functional cortex,which may be related to changes in their brain function.

Objective To explore the relationship between the inhibitory effect of quercetin on AC16 cardiomyocyte proliferation and the Wnt/β-catenin signaling pathway,the relationship between quercetin(quercetin,QCT)and the proliferation of AC16 cardiomyocytes through in vitro was investigated.Methods AC16 cells were stimulated with different concentrations of QCT.The effects of QCT on AC16 cell proliferation were detected by inverted microscope photography,trypan blue counting,and CCK-8 assay.Western blot was used to detect the effects of QCT on the expression of phosphorylated β-catenin(p-β-catenin),c-Myc,β-catenin,low density lipoprotein receptor-related protein 5(LRP5),and LRP6.The effect of quercetin on cell proliferation was detected after overexpressing β-catenin ΔN,LRP5,and LRP6 genes,and after silencing LRP5 and LRP6 genes by trypan blue counting.Results Compared with the control group,QCT could decrease the number of AC16 cells and inhibit the proliferation rate,which was concentration-dependent.At the protein expression level,10 and 20 μmol/L QCT led to an significant upregulated modification of p-β-catenin protein(P<0.05)and significant downregulation of c-Myc,β-catenin,LRP5,and LRP6 protein expression(P<0.05)in AC16 cells.Therefore,10 μmol/L QCT was chosen as the intervention concentration.Overexpression of β-catenin ΔN,LRP5,and LRP6 genes in AC16 cells significantly rescued the cell proliferation inhibition caused by 10 μmol/L QCT compared to the drug-only group(P<0.05).Conversely,silencing LRP5 and LRP6 genes led to inhibition of AC16 cell proliferation,and the combination with 10 μmol/L QCT did not exacerbate the inhibition(P>0.05).Conclusion The quercetin could inhibit the Wnt/β-catenin signaling pathway via significant downregulation of LRP5/6,thereby attenuate cell proliferation of AC16 cardiomyocytes.

Objective To investigate the anti-hepatocellular carcinoma mechanism of Chloranthus fortunei(A.Gray)Solms-Laub.via network pharmacology,molecular docking techniques and in vitro experiments.Methods Chemical composition of Chloranthus fortunei(A.Gray)Solms-Laub.was searched by literature.Swiss Target Prediction was used to find corresponding targets.STRING was used to construct protein-protein interactions network(PPI).DAVID was used to enrich GO analysis and KEGG pathway.AutoDock Vina 1.1.2 and Pymol visualisation was used for docking and validation.Results Chloranthus fortunei(A.Gray)Solms-Laub.had 61 active components,685 targets,and 279 intersections with disease targets.The PPI showed that the main active components were Luteolin,Chloranthalactone C,Shizukanolide H,Esculetin,7-Hydroxycoumarin.The key targets were GAPDH,VEGFA,STAT3,JUN,HSP90AA1,AKT1,CTNNB1,CASP3,and ALB.Biological process(BP)involved protein phosphorylation,signal transduction,regulation of RNA polymerase II promoter transcription,cell proliferation,apoptosis.Cellular component(CC)involved cytoplasm,nucleus,cell membrane,cellular exosome.Molecular function(MF)involves protein binding,ATPase,threonine kinase,protein kinase activity.KEGG involved cancer pathway,metabolic pathway,PI3K-Akt signalling pathway,cancer proteoglycans,lipids and atherosclerosis,cytomegalovirus infection,microRNAs in cancer,human T-cell leukaemia virus type 1,Ras signalling pathway,MAPK signalling pathway.Molecular docking showed that silverweed lactone H had a strong affinity for each of the other target proteins,indicating that this component plays a key role.The results of RT-qPCR assay and WB assay showed that there were significant differences in gene and protein expression levels before and after drug administration.Conclusion The Chinese medicine in Chloranthus fortunei(A.Gray)Solms-Laub.can treat hepatocellular carcinoma through the MAPK pathway,and the main active ingredients have good docking effects with the core target proteins of the disease.

Objective To investigate the value of whole lung CT radiomics combined with clinical and conventional CT features for differentiating non-tuberculous mycobacterial pulmonary disease(NTM-PD)and pulmonary tuberculosis(PTB).Methods Fifty-three NTM-PD(NTM-PD group)and 111 PTB(PTB group)patients diagnosed by mycobacteria culture were retrospectively collected.The patients were divided into training set(n=115,including 37 cases of NTM-PD and 78 cases of PTB)and test set(n=49,including 16 cases of NTM-PD and 33 cases of PTB)at the ratio of 7∶3.Patients'clinical and pulmonary CT manifestations of lesions were analyzed using univariate and multivariate logistic regression,and the independent impact factors for differentiating NTM-PD and PTB lesions were screened.The best radiomics features were extracted and screened based on whole lung CT.Based on independent impact factors,the best radiomics features and their combination,clinical-CT,radiomics and combined models were constructed with random forest,and the differential efficacy of each model was evaluated.Results Patients'age(OR=0.264),gender(OR=0.956),immunoglobulin G(OR=3.416),C reactive protein(OR=3.418)and bronchiectasis shown on CT(OR=0.285)were all independent impact factors for differentiating NTM-PD and PTB.Twelve best radiomics features were screened based on whole lung ROI.The AUC of combined model in training set and test set was 0.915 and 0.901,respectively,both higher than that of clinical model(AUC=0.832,0.801,Z=1.340,3.710,both P＜0.05)and radiomics model(AUC=0.877,0.821,Z=-2.520,-5.240,both P＜0.05).Conclusion Whole lung CT radiomics combined with clinical and conventional CT features could effectively distinguish NTM-PD and PTB.

Objective:To investigate the rates of low disease activity and clinical remission in patients with systemic lupus erythematosus(SLE)in a real-world setting,and to analyze the related factors of low disease activity and clinical remission.Methods:One thousand patients with SLE were enrolled from 11 teaching hospitals.Demographic,clinical and laboratory data,as well as treatment regimes were collec-ted by self-completed questionnaire.The rates of low disease activity and remission were calculated based on the lupus low disease activity state(LLDAS)and definitions of remission in SLE(DORIS).Charac-teristics of patients with LLDAS and DORIS were analyzed.Multivariate Logistic regression analysis was used to evaluate the related factors of LLDAS and DORIS remission.Results:20.7％of patients met the criteria of LLDAS,while 10.4％of patients achieved remission defined by DORIS.Patients who met LLDAS or DORIS remission had significantly higher proportion of patients with high income and longer disease duration,compared with non-remission group.Moreover,the rates of anemia,creatinine eleva-tion,increased erythrocyte sedimentation rate(ESR)and hypoalbuminemia was significantly lower in the LLDAS or DORIS group than in the non-remission group.Patients who received hydroxychloroquine for more than 12 months or immunosuppressant therapy for no less than 6 months earned higher rates of LLDAS and DORIS remission.The results of Logistic regression analysis showed that increased ESR,positive anti-dsDNA antibodies,low level of complement(C3 and C4),proteinuria,low household in-come were negatively related with LLDAS and DORIS remission.However,hydroxychloroquine usage for longer than 12 months were positively related with LLDAS and DORIS remission.Conclusion:LLDAS and DORIS remission of SLE patients remain to be improved.Treatment-to-target strategy and standar-dized application of hydroxychloroquine and immunosuppressants in SLE are recommended.