AIM:To investigate the mechanism of Qianggu-Kangshu formula(QGKSF)in the treatment of rheumatoid arthritis(RA)bone destruction based on network pharmacology,molecular docking,and cell experiments.METHODS:The main active ingredients and potential targets of QGKSF were obtained through TCMSP database and litera-ture search.OMIM database and GeneCards database were used to search the targets related to RA bone destruction,and Venny 2.1.0 was employed to screen the intersected targets of QGKSF and RA bone destruction.The protein-protein inter-action network of potential intersected targets was constructed by STRING database,and topological analysis was carried out by Cytoscape software to screen core targets.The screened targets of QGKSF related to RA bone destruction were evalu-ated through the Metascape system.Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology were applied to complete enrichment analysis.AutoDock and PyMOL software was used to carry out molecular docking between the core components and the core proteins.Mouse RAW264.7 macrophages were cultured in vitro,and the cell viability was de-tected by CCK-8 assay.The number of tartrate-resistant acid phosphatase(TRAP)positive multinucleated cells in each group was calculated by TRAP staining.The enzyme activity of the cells was evaluated by determination of TRAP activity.F-actin ring formation was detected by phalloidin staining.Western blot analysis was conducted to detect the protein levels of nuclear factor of activated T cells 1(NFATc1),TRAP,cathepsin K(CTSK),c-Fos,matrix metalloproteinase 9(MMP9),osteoprotegerin(OPG),receptor activator of nuclear factor κB(RANK),RANK ligand(RANKL)and phos-phorylated protein kinase B(p-AKT).RESULTS:A total of 136 active ingredients and 126 targets were selected corre-lated with QGKSF,whil 207 intersected targets of QGKSF and RA bone destruction were screened,175 of which were core targets.There were 199 pathways obtained by GO enrichment,and 20 pathways related to osteoclasts were screened out,including phosphatidylinoinosiol 3-kinase/AKT signaling pathway and osteoclast differentiation,etc.The results of TRAP staining,TRAP enzyme activity determination and phalloidin staining showed less positive cell formation,de-creased enzyme activity and decreased F-actin ring formation in QGKSF and methotrexate(MTX)groups compared with model group.Western blot results showed that compared with model group,the protein levels of NFATc1,TRAP,CTSK,c-Fos,MMP9,p-AKT,RANK and RANKL were decreased(P＜0.05),while the expression of OPG protein was in-creased(P＜0.05)in QGKSF and MTX groups.CONCLUSION:Treatment with QGKSF inhibits RANKL-induced dif-ferentiation of RAW264.7 cells into osteoclasts possibly by inhibiting the over-differentiation of osteoclasts via regulating RANKL/RANK/OPG signaling pathway.

AIM:To investigate the mechanism of Qianggu-Kangshu formula(QGKSF)in the treatment of rheumatoid arthritis(RA)bone destruction based on network pharmacology,molecular docking,and cell experiments.METHODS:The main active ingredients and potential targets of QGKSF were obtained through TCMSP database and litera-ture search.OMIM database and GeneCards database were used to search the targets related to RA bone destruction,and Venny 2.1.0 was employed to screen the intersected targets of QGKSF and RA bone destruction.The protein-protein inter-action network of potential intersected targets was constructed by STRING database,and topological analysis was carried out by Cytoscape software to screen core targets.The screened targets of QGKSF related to RA bone destruction were evalu-ated through the Metascape system.Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology were applied to complete enrichment analysis.AutoDock and PyMOL software was used to carry out molecular docking between the core components and the core proteins.Mouse RAW264.7 macrophages were cultured in vitro,and the cell viability was de-tected by CCK-8 assay.The number of tartrate-resistant acid phosphatase(TRAP)positive multinucleated cells in each group was calculated by TRAP staining.The enzyme activity of the cells was evaluated by determination of TRAP activity.F-actin ring formation was detected by phalloidin staining.Western blot analysis was conducted to detect the protein levels of nuclear factor of activated T cells 1(NFATc1),TRAP,cathepsin K(CTSK),c-Fos,matrix metalloproteinase 9(MMP9),osteoprotegerin(OPG),receptor activator of nuclear factor κB(RANK),RANK ligand(RANKL)and phos-phorylated protein kinase B(p-AKT).RESULTS:A total of 136 active ingredients and 126 targets were selected corre-lated with QGKSF,whil 207 intersected targets of QGKSF and RA bone destruction were screened,175 of which were core targets.There were 199 pathways obtained by GO enrichment,and 20 pathways related to osteoclasts were screened out,including phosphatidylinoinosiol 3-kinase/AKT signaling pathway and osteoclast differentiation,etc.The results of TRAP staining,TRAP enzyme activity determination and phalloidin staining showed less positive cell formation,de-creased enzyme activity and decreased F-actin ring formation in QGKSF and methotrexate(MTX)groups compared with model group.Western blot results showed that compared with model group,the protein levels of NFATc1,TRAP,CTSK,c-Fos,MMP9,p-AKT,RANK and RANKL were decreased(P＜0.05),while the expression of OPG protein was in-creased(P＜0.05)in QGKSF and MTX groups.CONCLUSION:Treatment with QGKSF inhibits RANKL-induced dif-ferentiation of RAW264.7 cells into osteoclasts possibly by inhibiting the over-differentiation of osteoclasts via regulating RANKL/RANK/OPG signaling pathway.

Objective To compare the bone mineral density and age-related bone loss rate of thoracic and lumbar vertebrae using quantitative CT(QCT),and to explore the feasibility of measuring bone density and bone loss rate using thoracic QCT.Methods A total of 708 patients who underwent full abdominal CT scans due to trauma,acute abdominal disease,and health examination from March 2022 to March 2023 were selected.The average BMD of T10 to L3 vertebral bodies were measured using quantitative CT software.The subjects were divided into several age groups according to the age of every 10 years.The average bone density and the age-related annual rate of bone density loss of the thoracic and lumbar vertebrae for each group were computed,the correlation analysis between thoracic and lumbar BMD and age was conducted using Pearson correlation analysis.Results The peak BMD for both males and females was in the 20 to 29 years age group.After the age of 30,the bone density of the thoracic and lumbar vertebrae began to decrease to varying degrees.For females aged 80 to 89,the cumulative bone loss rate for the thoracic and lumbar vertebrae was 60.61%and 61.34%,respectively,while for males,it was 44.45%and 49.35%,respectively.The BMD of the thoracic and lumbar vertebrae in both males and females showed a negative correlation with age(P<0.01).There was a significant positive correlation between the bone mineral density of the thoracic and lumbar vertebrae(for males,r = 0.96;for females,r = 0.98;P<0.01).Conclusion Both thoracic and lumbar QCT accurately displayed BMD and bone loss in different age groups,and thoracic QCT could be used as a method for evaluating and monitoring bone loss.

Objective To explore the diagnostic value of dual syringe and dual-channel direct multi-slice computed tomography venography(MSCTV) for lower extremity deep venous obstructive disease.Methods 100 patients highly suspected deep vein thrombosis by clinic wereunderwent dual syringe and dual-channel direct MSCTV.The original data processed with technology of subtraction,and put into workstation and reconstructed with maximum intensity projection(MIP),multiplanar reformation(MPR) and volume rendering (VR).Images of deep venous obstructive lesions were analysed and graded.All patients were underwent DSA in one week.Results Direct MSCTV showed that the reconstructed images and the vascular contrast were very clear.In the evaluation of deep venous,excellent proportion was 95 ％ for the inferior vena cava,94 ％ for the common iliac vein,95 ％ for the external iliac vein,95.5 ％ for the femoral vein,96.5％ for the popliteal vein and 92 ％ for the low leg vein.Direct MSCTV displayed complete and regular vein in 8 normal patients,92 cases displayed different parts and different degrees of thrombosis.On original axial images,eccentric filling defect of vascular were showed (there were 64 multiple vein occlusion cases and 28 solitary obstruction cases).The occlusion locations in deep venous were as following:3 in the inferior vena cava,67 in the common iliac vein,28 in the external iliac vein,50 in the femoral vein,26 in the popliteal vein,89 in the anterior tibial vein,35 in the posterior tibial vein and 5 in the peroneal vein.There were 7 normal patients and 93 patients with different parts and different degrees of thrombosis.Conclusion Direct MSCTV can accurately show deep vein thrombosis,which image is clear and reliable for displaying the scope and extent of lesions of the vascular and whether the collateral circulation established.Direct MSCTV has a significant clinical value in diagnosing the thrombotic disease of deep vein obstructive lesions.

Clopidogrel was believed to be superior to aspirin by the well-known CAPRIE trial. However, no other large clinical trials demonstrated the same results, but all focused on the combination use of clopidogrel with aspirin, and combination therapy in CREDO was called the "Emperor's New Clothes". However, no one overturned the results of these clinical trials by quantitatively analyzing them. We reviewed ten large-scale clinical trials about clopidogrel. On the basis of results of CAPRIE, CREDO and CHARISMA trials, we re-estimated their minimal sample sizes and their powers by three well-established statistical methodologies. From the results of CAPRIE, we inferred that the minimal sample size should be 85 086 or 84 968 but its power was only 30.70%. A huge gap existed. The same was also true of CREDO and CHARISMA trials. Moreover, in CAPRIE trial, 0 was included in the 95% confidence interval and 1 was included in the 95% confidence interval for the relative risk. There were some paradoxical data in CAPRIE trial. We are led to conclude that the results in CAPRIE, CREDO, and from the subgroup analysis in CHARISMA trials were questionable. These results failed to demonstrate that clopidogrel was superior to aspirin or that clopidogrel used in combination with aspirin was better than aspirin alone. The cost-effectiveness analyses by some previous studies were not reliable.

Clopidogrel was believed to be superior to aspirin by the well-known CAPRIE trial. However, no other large clinical trials demonstrated the same results, but all focused on the combina-tion use of clopidogrel with aspirin, and combination therapy in CREDO was called the "Emperor's New Clothes". However, no one overturned the results of these clinical trials by quantitatively ana-lyzing them. We reviewed ten large-scale clinical trials about clopidogrel. On the basis of results of CAPRIE, CREDO and CHARISMA trials, we re-estimated their minimal sample sizes and their powers by three well-established statistical methodologies. From the results of CAPRIE, we inferred that the minimal sample size should be 85 086 or 84 968 but its power was only 30.70%. A huge gap existed. The same was also true of CREDO and CHARISMA trials. Moreover, in CAPRIE trial, 0 was included in the 95% confidence interval and 1 was included in the 95% confidence interval for the relative risk. There were some paradoxical data in CAPRIE trial. We are led to conclude that the results in CAPRIE, CREDO, and from the subgroup analysis in CHARISMA trials were questionable. These results failed to demonstrate that clopidogrel was superior to aspirin or that clopidogrel used in combination with aspirin was better than aspirin alone. The cost-effectiveness analyses by some pre-vious studies were not reliable.