目的分析细针穿刺细胞学检查(FNAC)BethesdaⅢ类甲状腺结节结合组织Galectin-3分子表达和超声造影(CEUS)在鉴别良恶性中的应用价值。方法选择行FNAC检查诊断为BethesdaⅢ类甲状腺结节患者110例(140个结节),其中男性52例,女性58例;年龄25...目的分析细针穿刺细胞学检查(FNAC)BethesdaⅢ类甲状腺结节结合组织Galectin-3分子表达和超声造影(CEUS)在鉴别良恶性中的应用价值。方法选择行FNAC检查诊断为BethesdaⅢ类甲状腺结节患者110例(140个结节),其中男性52例,女性58例;年龄25~64岁,平均年龄45.5岁。所有组织分别进行Galectin-3分子免疫组织化学染色和CEUS两项检查,诊断结节的良恶性,比较良性组和恶性组常规超声甲状腺影像报告与数据系统(TI-RADS)分级、Galectin-3表达阳性率、CEUS增强模式及CEUS定量参数。采用受试者工作特性(ROC)曲线评估鉴别恶性结节的准确度。结果经细胞病理学确诊良性组患者80例,良性结节100个;恶性组患者30例,恶性结节40个。恶性组与良性组患者性别、年龄、TI-RADS分级、结节直径比较差异无统计学意义(P>0.05)。恶性组结节Galectin-3表达阳性率显著高于良性组(60.0%vs 30.0%),CEUS增强模式以早期低增强为主,CEUS定量评估软件测量结节峰值强度(PI)值低于良性组(5.32±1.46 vs 6.79±1.88)(χ^(2)=10.853、25.239,t=4.968,P<0.05)。以Galectin-3表达阳性联合CEUS检查作为评估BethesdaⅢ类甲状腺结节良恶性的主要依据,手术病理诊断结果为金标准,得出诊断恶性结节的准确度61.82%(34/55),灵敏度85.00%(34/40),特异度79.00%(79/100),阳性预测值61.82%(34/55)和阴性预测值92.94%(79/85)。采用ROC曲线进一步验证联合Galectin-3表达阳性和CEUS检查诊断BethesdaⅢ类甲状腺结节良恶性,结果准确度为82.5%,灵敏度80.9%,特异度75.6%。结论CEUS定量检查联合结节Galectin-3分子表达能够为FNAC诊断BethesdaⅢ类甲状腺结节的良恶性质提供更加丰富的生物学信息,CEUS增强模式以早期低增强为主,Galectin-3阳性表达有助于结节恶性的判断。展开更多
Objective:The main characteristics of diabetic nephropathy(DN)at the early stage are abnormal angiogenesis of glomerular endothelial cells(GECs)and macrophage infiltration.Galectin-3 plays a pivotal role in the pathog...Objective:The main characteristics of diabetic nephropathy(DN)at the early stage are abnormal angiogenesis of glomerular endothelial cells(GECs)and macrophage infiltration.Galectin-3 plays a pivotal role in the pathogenesis of DN via binding with its ligand,advanced glycation end products(AGEs).Catalpol,an iridoid glucoside extracted from Rehmannia glutinosa,has been found to ameliorate vascular inflammation,reduce endothelial permeability,and protect against endothelial damage in diabetic milieu.However,little is known about whether catalpol could exert an anti-angiogenesis and anti-inflammation effect induced by AGEs.Methods:Mouse GECs(mGECs)and RAW 264.7 macrophages were treated with different concentrations of AGEs(0,50,100,200 and 400μg/mL)for different time(0,6,12,24 and 48 h)to determine the optimal concentration of AGEs and treatment time.Cells were treated with catalpol(10μmol/L),GB1107(1μmol/L,galectin-3 inhibitor),PX-478(50μmol/L,HIF-1αinhibitor),adenovirus-green fluorescent protein(Ad-GFP)[3×10^(7)plaque-forming unit(PFU)/mL]or Ad-galectin-3-GFP(2×10^(8)PFU/mL),which was followed by incubation with 50μg/mL AGEs.The levels of galectin-3,vascular endothelial growth factor A(VEGFA)and pro-angiogenic factors angiopoietin-1(Ang-1),angiopoietin-2(Ang-2),tunica interna endothelial cell kinase-2(Tie-2)were detected by enzyme-linked immunosorbent assay(ELISA).Cell counting kit-8(CCK-8)assay was used to evaluate the proliferation of these cells.The expression levels of galectin-3,vascular endothelial growth factor receptor 1(VEGFR1),VEGFR2,and hypoxia-inducible factor-1α(HIF-1α)in mGECs and those of galectin-3 and HIF-1αin RAW 264.7 macrophages were detected by Western blotting and immunofluorescence(IF)staining.The rat DN model was established.Catalpol(100 mg/kg)or GB1107(10 mg/kg)was administered intragastrically once a day for 12 weeks.Ad-galectin-3-GFP(6×10^(7)PFU/mL,0.5 mL)or Ad-GFP(6×10^(6)PFU/mL,0.5 mL)was injected into the tail vein of rats 48 h before the sacrifice of the animals.The expression of galectin-3,VEGFR1,.VEGFR2,and HIF-1αin renal cortices was analyzed by Western blotting.The expression of galectin-3,F4/80(a macrophage biomarker),and CD34(an endothelium biomarker)in renal cortices was detected by IF staining,and collagen accumulation by Masson staining.Results:The expression levels of galectin-3 and VEGFA were significantly higher in mGECs and RAW 264.7 macrophages treated with 50μg/mL AGEs for 48 h than those in untreated cells.Catalpol and GB1107 could block the AGEs-induced proliferation of mGECs and RAW 264.7 macrophages.Over-expression of galectin-3 was found to reduce the inhibitory effect of catalpol on the proliferation of cells.Catalpol could significantly decrease the levels of Ang-1,Ang-2 and Tie-2 released by AGEs-treated mGECs,which could be reversed by over-expression of galectin-3.Catalpol could significantly inhibit AGEs-induced expression of galectin-3,HIF-1α,VEGFR1,and VEGFR2 in mGECs.The inhibitory effect of catalpol on galectin-3 in AGEs-treated mGECs was impaired by PX-478.Moreover,catalpol attenuated the AGEs-activated HIF-1α/galectin-3 pathway in RAW 264.7 macrophages,which was weakened by PX-478.Additionally,catalpol significantly inhibited the expression of galectin-3,macrophage infiltration,collagen accumulation,and angiogenesis in the kidney of diabetic rats.Over-expression of galectin-3 could antagonize these inhibitory effects of catalpol.Conclusion:Catalpol prevented the angiogenesis of mGECs and macrophage proliferation via inhibiting galectin-3.It could prevent the progression of diabetes-induced renal damage.展开更多
基金Key Program of Natural Science Foundation of Shenzhen(JCYJ20220818102218039)Shenzhen Science and Technology Program(KCXFZ20230731093559005)+2 种基金Natural Science Foundation of Shenzhen(JCYJ20210324133412033)Guangdong Province Innovation Team Project for Universities(2023KCXTD049)Shenzhen Key Medical Discipline Construction Fund(SZXK045)。
文摘目的分析细针穿刺细胞学检查(FNAC)BethesdaⅢ类甲状腺结节结合组织Galectin-3分子表达和超声造影(CEUS)在鉴别良恶性中的应用价值。方法选择行FNAC检查诊断为BethesdaⅢ类甲状腺结节患者110例(140个结节),其中男性52例,女性58例;年龄25~64岁,平均年龄45.5岁。所有组织分别进行Galectin-3分子免疫组织化学染色和CEUS两项检查,诊断结节的良恶性,比较良性组和恶性组常规超声甲状腺影像报告与数据系统(TI-RADS)分级、Galectin-3表达阳性率、CEUS增强模式及CEUS定量参数。采用受试者工作特性(ROC)曲线评估鉴别恶性结节的准确度。结果经细胞病理学确诊良性组患者80例,良性结节100个;恶性组患者30例,恶性结节40个。恶性组与良性组患者性别、年龄、TI-RADS分级、结节直径比较差异无统计学意义(P>0.05)。恶性组结节Galectin-3表达阳性率显著高于良性组(60.0%vs 30.0%),CEUS增强模式以早期低增强为主,CEUS定量评估软件测量结节峰值强度(PI)值低于良性组(5.32±1.46 vs 6.79±1.88)(χ^(2)=10.853、25.239,t=4.968,P<0.05)。以Galectin-3表达阳性联合CEUS检查作为评估BethesdaⅢ类甲状腺结节良恶性的主要依据,手术病理诊断结果为金标准,得出诊断恶性结节的准确度61.82%(34/55),灵敏度85.00%(34/40),特异度79.00%(79/100),阳性预测值61.82%(34/55)和阴性预测值92.94%(79/85)。采用ROC曲线进一步验证联合Galectin-3表达阳性和CEUS检查诊断BethesdaⅢ类甲状腺结节良恶性,结果准确度为82.5%,灵敏度80.9%,特异度75.6%。结论CEUS定量检查联合结节Galectin-3分子表达能够为FNAC诊断BethesdaⅢ类甲状腺结节的良恶性质提供更加丰富的生物学信息,CEUS增强模式以早期低增强为主,Galectin-3阳性表达有助于结节恶性的判断。
基金supported by grants from the National Natural Science Foundation of China(No.81374029,No.81073111,No.81874359)Natural Science Foundation of the Higher Education Institutions of Jiangsu Province(No.18KJD360002)+1 种基金a Project Funded by Jiangsu Agri-animal Husbandry Vocational College(No.NSF2021CB04)a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)(No.JKLPSE201604).
文摘Objective:The main characteristics of diabetic nephropathy(DN)at the early stage are abnormal angiogenesis of glomerular endothelial cells(GECs)and macrophage infiltration.Galectin-3 plays a pivotal role in the pathogenesis of DN via binding with its ligand,advanced glycation end products(AGEs).Catalpol,an iridoid glucoside extracted from Rehmannia glutinosa,has been found to ameliorate vascular inflammation,reduce endothelial permeability,and protect against endothelial damage in diabetic milieu.However,little is known about whether catalpol could exert an anti-angiogenesis and anti-inflammation effect induced by AGEs.Methods:Mouse GECs(mGECs)and RAW 264.7 macrophages were treated with different concentrations of AGEs(0,50,100,200 and 400μg/mL)for different time(0,6,12,24 and 48 h)to determine the optimal concentration of AGEs and treatment time.Cells were treated with catalpol(10μmol/L),GB1107(1μmol/L,galectin-3 inhibitor),PX-478(50μmol/L,HIF-1αinhibitor),adenovirus-green fluorescent protein(Ad-GFP)[3×10^(7)plaque-forming unit(PFU)/mL]or Ad-galectin-3-GFP(2×10^(8)PFU/mL),which was followed by incubation with 50μg/mL AGEs.The levels of galectin-3,vascular endothelial growth factor A(VEGFA)and pro-angiogenic factors angiopoietin-1(Ang-1),angiopoietin-2(Ang-2),tunica interna endothelial cell kinase-2(Tie-2)were detected by enzyme-linked immunosorbent assay(ELISA).Cell counting kit-8(CCK-8)assay was used to evaluate the proliferation of these cells.The expression levels of galectin-3,vascular endothelial growth factor receptor 1(VEGFR1),VEGFR2,and hypoxia-inducible factor-1α(HIF-1α)in mGECs and those of galectin-3 and HIF-1αin RAW 264.7 macrophages were detected by Western blotting and immunofluorescence(IF)staining.The rat DN model was established.Catalpol(100 mg/kg)or GB1107(10 mg/kg)was administered intragastrically once a day for 12 weeks.Ad-galectin-3-GFP(6×10^(7)PFU/mL,0.5 mL)or Ad-GFP(6×10^(6)PFU/mL,0.5 mL)was injected into the tail vein of rats 48 h before the sacrifice of the animals.The expression of galectin-3,VEGFR1,.VEGFR2,and HIF-1αin renal cortices was analyzed by Western blotting.The expression of galectin-3,F4/80(a macrophage biomarker),and CD34(an endothelium biomarker)in renal cortices was detected by IF staining,and collagen accumulation by Masson staining.Results:The expression levels of galectin-3 and VEGFA were significantly higher in mGECs and RAW 264.7 macrophages treated with 50μg/mL AGEs for 48 h than those in untreated cells.Catalpol and GB1107 could block the AGEs-induced proliferation of mGECs and RAW 264.7 macrophages.Over-expression of galectin-3 was found to reduce the inhibitory effect of catalpol on the proliferation of cells.Catalpol could significantly decrease the levels of Ang-1,Ang-2 and Tie-2 released by AGEs-treated mGECs,which could be reversed by over-expression of galectin-3.Catalpol could significantly inhibit AGEs-induced expression of galectin-3,HIF-1α,VEGFR1,and VEGFR2 in mGECs.The inhibitory effect of catalpol on galectin-3 in AGEs-treated mGECs was impaired by PX-478.Moreover,catalpol attenuated the AGEs-activated HIF-1α/galectin-3 pathway in RAW 264.7 macrophages,which was weakened by PX-478.Additionally,catalpol significantly inhibited the expression of galectin-3,macrophage infiltration,collagen accumulation,and angiogenesis in the kidney of diabetic rats.Over-expression of galectin-3 could antagonize these inhibitory effects of catalpol.Conclusion:Catalpol prevented the angiogenesis of mGECs and macrophage proliferation via inhibiting galectin-3.It could prevent the progression of diabetes-induced renal damage.
