BACKGROUND:Pinoresinol diglucoside promotes bone formation and bone matrix synthesis and accelerates bone tissue repair.However,the mechanism of action and effects of this compound in osteoblasts need to be further explored. OBJECTIVE:To investigate the effect and mechanism of action of pinoresinol diglucoside on dexamethasone-treated osteoblasts based on the Wnt/β-catenin signaling pathway. METHODS:Different concentrations of dexamethasone groups and pinoresinol diglucoside groups were set to treat osteoblasts for 24 hours,and the optimal intervention concentrations were screened.Osteoblasts were treated with dexamethasone,pinoresinol diglucoside and inhibitor XAV-939.Then,control group,dexamethasone group,XVA-939 group,pinoresinol diglucoside group,pinoresinol diglucoside+XVA-939 group were set up.Cell counting kit-8 assay was used to detect cell activity.Alkaline phosphatase activity and caspase3/7 enzyme activity in cells were detected.Annexin V/PI staining and EdU assay were used to detect cell apoptosis and proliferation.Real-time qPCR and western blot were used to detect the mRNA and protein expression levels of Wnt3a,β-catenin,c-myc,osteocalcin,and type I collagen,respectively. RESULTS AND CONCLUSION:After dexamethasone and pinoresinol diglucoside intervened in osteoblasts for 24 hours,10 μmol/L dexamethasone was found to be the optimal intervention concentration for cell inhibition,and cell proliferation was most pronounced at a concentration of pinoresinol diglucoside of 100 μmol/L.Compared with the dexamethasone group,alkaline phosphatase activity was significantly enhanced(P<0.05)and caspase3/7 enzyme activity was significantly reduced(P<0.05)in the pinoresinol diglucoside group.Annexin V/PI staining and cell proliferation assay by EdU method showed that pinoresinol diglucoside inhibited apoptosis and promoted proliferation of osteoblasts after dexamethasone intervention.The mRNA and protein expression levels of Wnt3a,β-catenin,c-myc,osteocalcin,and type I collagen were significantly higher in the pinoresinol diglucoside group and pinoresinol diglucoside+XVA-939 group compared with the dexamethasone and XVA-939 groups(P<0.05).To conclude,pinoresinol diglucoside can inhibit osteoblast apoptosis after dexamethasone intervention,protect osteoblast activity and promote osteoblast proliferation by activating the Wnt/β-catenin signaling pathway,which may play a role in delaying steroid-induced osteonecrosis of the femoral head.

Objective:To investigate the effect of icariin(ICA)on chondrocyte function in osteoarthritis(OA)and its mechanism.Methods:After chondrocytes were treated with different concentrations of ICA(0.1,0.5,1.0,5.0,10.0 μmol/L),methylthiazolydiphenyl-tetrazolium(MTT)assay was used to measure the viability of chondrocytes,and then the optimal concentration of ICA was selected for subsequent experiments.Chondrocytes were stimulated by interleukin-1β(IL-1β,10 ng/mL)for 24 hours to establish a cell model of OA,and then the cells were treated with ICA at the optimal concentration of 1 μmol/L.Quantitative real-time polymerase chain reac-tion(RT-qPCR)was used to measure the expression level of mi-croRNA-29c(miR-29c);Western blot was used to measure the pro-tein expression level of collagen type Ⅱ alpha 1 chain(COL2A1);Enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of matrix metalloproteinase-3(MMP-3)and tumor necro-sis factor-α(TNF-α)in the supernatant of cell culture;flow cytom-etry was used to measure cell apoptosis rate.Bioinformatics web-sites were used to predict the target genes of miR-29c,and dual-luciferase reporter assay was used to verify the targeted binding be-tween miR-29c and COL2A1.Functional rescue experiments were used to clarify the effect of ICA on inflammatory response and cell apoptosis in OA and its mechanism of action.Results:Compared with the control group,the IL-1β group had significant increases in the content of MMP-3(P<0.001)and TNF-α(P<0.001)in chondrocyte supernatant,a significant reduction in the expression level of COL2A1(P<0.001),and a significant increase in cell apoptosis rate(P<0.001).Compared with the IL-1β group,the ICA treatment group had significant reductions in the content of MMP-3(P=0.001;P<0.001)and TNF-α(P<0.001;P<0.001)in chondrocyte super-natant,a significant increase in the expression level of COL2A1(P<0.001;P<0.001),and a significant reduction in cell apoptosis rate(P<0.001;P<0.001).Compared with the control group,the IL-1β group had a significant increase in the expression level of miR-29c in chondrocytes(P<0.001),while ICA treatment significantly reduced the expression level of miR-29c(P<0.001).Compared with the control group,the miR-29c mimic group had significant increases in the content of MMP-3(P<0.001)and TNF-α(P<0.001)in chon-drocyte supernatant,a significant reduction in the expression level of COL2A1(P=0.001),and a significant increase in cell apoptosis rate(P<0.001);however,opposite results were obtained for the miR-29c inhibitor group.TargetScan7.1 database showed the exis-tence of binding sites between miR-29c and COL2A1.Dual-luciferase reporter assay showed that miR-29c mimic significantly inhib-ited the luciferase activity of wild-type COL2A1 vector(P<0.001),but with no influence on the luciferase activity of mutant COL2A1 vector(P=0.140).Knockdown of COL2A1 could partially reverse the effect of downregulated miR-29c on the secretion of MMP-3(P<0.001)and TNF-α(P<0.001),the protein expression level of COL2A1(P<0.001),and cell apoptosis rate(P<0.001)in chondro-cytes.ICA treatment could alleviate the influence of miR-29c overexpression on the secretion of MMP-3(P<0.001)and TNF-α(P<0.001),the protein expression level of COL2A1(P<0.001),and the apoptosis rate(P<0.001)of chondrocytes.Conclusion:ICA can alleviate the secretion of inflammatory cytokines and the apoptosis of chondrocytes induced by IL-1β,which may be associated with the miR-29c/COL2A1 axis..

Objective:To compare carotid endarterectomy (CEA) and carotid artery stenting (CAS) in perioperative, medium and long term prognosis of patients with carotid artery stenosis.Methods:A retrospective analysis was performed on 1 329 cases of carotid artery stenosis treated at Department of Vascular Surgery, Beijing Anzhen Hospital from Jan 2011 to Aug 2020, as all cases being divided into CAS group and CEA group.Results:There were significant differences in age ( t=0.098, P=0.023) and drinking habits ( χ2=8.055, P=0.005) between the two groups. There were more unstable plaques in CEA group ( χ2=4.392, P=0.038), and more bilateral lesions in CAS group ( χ2=9.673, P=0.038). In perioperative period, there were more mannitol use in CEA group ( χ2=78.614, P<0.001), more incision/puncture site complications ( χ2=5.158, P=0.035), lung infection ( χ2=6.355, P=0.013), cerebral hyperperfusion syndrome (CHS) ( χ2=5.158, P=0.035) and extracranial nerve injury ( χ2=23.760, P<0.001) in CEA group than in CAS group, and more acute renal failure in CAS group ( χ2=10.393, P=0.001). There was no significant difference in survival rate and ischemic stroke, myocardial infarction, cerebral hemorrhage and renal insufficiency between the two groups (all P>0.05). The mean survival time of CAS group was 53.195 months (95% CI: 52.040-54.350), and 54.492 months (95% CI: 53.790-55.195) in CEA group ( P=0.051). Conclusions:Patients in CEA group had more unstable plaque and a lower perioperative stroke rate. CEA group had higher risk of CHS,while CAS was with lower postoperative lung infection rate and less wound local complications. There was no significant difference in long-term survival between the two groups.

Objective To analyze the outcome of intra-artery chemotherapy for T1G3 bladder cancer , and its effectiveness and safety. Methods From June 2003 to May 2014, 39 patients with T1G3 bladder cancer chose intra-artery chemotherapy (Gemcitabine plus cis-platin), and close follow-up was required after 2 cycles of chemotherapy. During the follow-up, transurethral resection of bladder tumor was performed for non-muscle invasive bladder cancer, and cystectomy was performed for muscle invasive tumor. Results Of all patients, 32 were male and 7 were female. The median age was 56 years old (range: 32-82 years), and median follow-up time was 56 months (range: 12-136 months). Nineteen patients were primary bladder cancer, and 20 were recurrent tumor. During the follow-up, 17 patients developed recurrent tumors, including 8 progressed tumors and 3 died from tumor. Two-year and 5-year progressed-free survival were 88% and 74%, and 2-year and 5-year cancer-specific survival were 97% and 89%, respectively. During 5 years′ follow-up, 81% survivor preserved intact bladder, and only 1 patient cancelled chemotherapy for adverse effect. Conclusions Intra-artery chemotherapy (GC regimen) is a choice for T1G3bladder cancer, preventing from disease progression with good tolerance.