Purpose: To evaluate the accuracy of Canon KU-1 IOL measurer (Japanese Canon Company) and VPLUS A/B scanner (French Quantel Company) in axial length (AL)measurement.Methods:Canon KU-1 IOL measurer and VPLUS A/B scanne...Purpose: To evaluate the accuracy of Canon KU-1 IOL measurer (Japanese Canon Company) and VPLUS A/B scanner (French Quantel Company) in axial length (AL)measurement.Methods:Canon KU-1 IOL measurer and VPLUS A/B scanner were used to measure axial length of human cataractous eyes before cataract surgery. Two hundred and twentytwo cases (433 eyes) were involved. The results were compared and the postoperative visual acuity, refractive results were recorded during the follow-ups to evaluate the accuracy of the two instruments.Results:In the 222 cases (433 eyes), the absolute value of the measurement differences was 0.4 mm or above in 35 eyes, 0.8 mm or above in 17 eyes, 1.2 mm or above in 12 eyes,2.0mm or above in 5 eyes. The refractive error was less than 2.0D in all patients. The mean values of ocular axial length by the two methods were 23.82 mm and 23.83 mm respectively and the difference had no statistic significance with compared t test ( P=0.902, two tail, or=0.01).Conclusion:The accurate AL measurements can be obtained with the two instruments and the measurement results should be analyzed comprehensively to obtain accurate values in the complicated cases.展开更多
Objectives:Mechanistic target of rapamycin(mTOR)activation has been identified in keloid.This study aimed to identify the role of mTOR-dependent autophagy activity in keloid.Methods:We detected the expression of speci...Objectives:Mechanistic target of rapamycin(mTOR)activation has been identified in keloid.This study aimed to identify the role of mTOR-dependent autophagy activity in keloid.Methods:We detected the expression of specific proteins representing mTOR activity and baseline autophagy levels in keloid tissues(KTs)and primary keloid fibroblasts(KFs)using immunohistochemical staining and western blotting.Simultaneously,the formation of acid vesicles was assessed by acridine orange staining in KFs.To investigate whether mTOR-dependent pathway mediated the regulation of autophagy machinery in keloid,we first validated whether mTOR inhibitors,rapamycin(100 nmol/L)and KU-0063794(5mmol/L),could inhibit mTOR activity in KFs by western blotting.Then we explored the reverse effects on autophagy activity induced by mTOR inhibitors in the presence of lysosomal protease inhibitors by western blotting.Results:It demonstrated elevated expression of mTOR,S6,and their activated forms in KTs,and an elevated expression of p-S6 Ser235/236 in KFs,suggesting mTOR was activated in keloid.Less LC3 and Beclin1 were expressed in the cytoplasm of KFs,whereas Ubiquitin was abundantly expressed in KTs compared with extra-lesional tissues.In addition,at the cellular level,an impeded conversion of LC3-I to LC3-II was shown in KFs and the formation of acid vesicles were also decreased in KFs compared with normal fibroblasts(NFs),indicating that autophagy activity is defective in keloid.mTOR inhibitors,Rapamycin(E-64d+pepstatin vs.rapamycin+E-64d+pepstatin:[0.88±0.35]vs.[1.56±0.46],F=5.56,P=0.049)and KU-0063794(E-64d+pepstatin vs.KU-0063794+E-64d+pepstatin:[0.92±0.22]vs.[1.51±0.25],F=25.88,P=0.011)can reverse the inhibition effect on autophagy of KFs while inhibiting mTOR activity.Conclusion:Autophagy machinery is inhibited in keloid which is regulated by mTOR-dependent pathway.展开更多
Objective:Blocking mechanistic target of rapamycin(mTOR)activation with mTOR inhibitors has promising therapeutic potential for keloids.However,the precise mechanism of mTOR inhibitors remains unclear.This study was a...Objective:Blocking mechanistic target of rapamycin(mTOR)activation with mTOR inhibitors has promising therapeutic potential for keloids.However,the precise mechanism of mTOR inhibitors remains unclear.This study was aimed to investigate the role of autophagy machinery in the anti-keloid effects of mTOR inhibitors.Methods:We first validated the biological effects induced by the mTOR inhibitors rapamycin(100 nmol/L)and KU-0063794(5μmol/L)on the proliferation,apoptosis,migration,and collagen synthesis of keloid fibroblasts(KFs)derived from Han Chinese persons through a Cell Counting Kit-8 assay,5-Bromo-2’-deoxyuridine incorporation,Annexin V/propidium iodide staining,migration,and western blotting.To explore whether autophagy machinery is involved in the anti-keloid effects of mTOR inhibitors,we first blocked the autophagy activation induced by rapamycin and KU-0063794 with a pharmacological autophagy inhibitor(wortmannin)or by silencing the key autophagy gene(ATG5),and we then re-evaluated these biological effects on KFs.Results:Blocking mTOR activation with either rapamycin or KU-0063794 completely inhibited proliferation,migration,and collagen synthesis of primary KFs but did not affect apoptosis.Incubating KFs with the autophagy inhibitor wortmannin or performingATG5 silencing abrogated the subsequent activation of autophagic activity induced by rapamycin(rapamycin+E-64d+pepstatinvs.rapamycin+wortmannin+E-64d+pepstatin:1.88±0.38vs.1.02±0.35,F=6.86,P=0.013),(non-sense control+rapamycinvs.ATG5 small interfering RNA+rapamycin:1.46±0.18vs.0.75±0.20,respectively;F=7.68,P=0.01)or KU-0063794(KU-0063794+E-64d+pepstatinvs.KU-0063794+wortmannin+E-64d+pepstatin:1.65±0.35vs.0.76±0.17,F=10.01,P=0.004),(NC+KU-0063794vs.ATG5 small interfering RNA+KU-0063794:1.59±0.50vs.0.77±0.09,F=5.93,P=0.02)as evidenced by decreased accumulation of LC3-II.However,blockage of autophagy induction in mTOR inhibitor-treated KFs with both methods did not disturb their anti-keloid effects,such as inhibition of cell viability,cell migration,and collagen synthesis(P>0.05 each).Conclusion:Blocking mTOR activation with the mTOR inhibitors rapamycin and KU-0063794 showed anti-keloid effects in KFs.Restoration of autophagy inhibition by mTOR inhibitors does not contribute to their anti-keloid effects.展开更多
文摘Purpose: To evaluate the accuracy of Canon KU-1 IOL measurer (Japanese Canon Company) and VPLUS A/B scanner (French Quantel Company) in axial length (AL)measurement.Methods:Canon KU-1 IOL measurer and VPLUS A/B scanner were used to measure axial length of human cataractous eyes before cataract surgery. Two hundred and twentytwo cases (433 eyes) were involved. The results were compared and the postoperative visual acuity, refractive results were recorded during the follow-ups to evaluate the accuracy of the two instruments.Results:In the 222 cases (433 eyes), the absolute value of the measurement differences was 0.4 mm or above in 35 eyes, 0.8 mm or above in 17 eyes, 1.2 mm or above in 12 eyes,2.0mm or above in 5 eyes. The refractive error was less than 2.0D in all patients. The mean values of ocular axial length by the two methods were 23.82 mm and 23.83 mm respectively and the difference had no statistic significance with compared t test ( P=0.902, two tail, or=0.01).Conclusion:The accurate AL measurements can be obtained with the two instruments and the measurement results should be analyzed comprehensively to obtain accurate values in the complicated cases.
基金This work was supported by CAMS Innovation Fund for Medical Sciences(CIFMS-2017-I2M-1-017)。
文摘Objectives:Mechanistic target of rapamycin(mTOR)activation has been identified in keloid.This study aimed to identify the role of mTOR-dependent autophagy activity in keloid.Methods:We detected the expression of specific proteins representing mTOR activity and baseline autophagy levels in keloid tissues(KTs)and primary keloid fibroblasts(KFs)using immunohistochemical staining and western blotting.Simultaneously,the formation of acid vesicles was assessed by acridine orange staining in KFs.To investigate whether mTOR-dependent pathway mediated the regulation of autophagy machinery in keloid,we first validated whether mTOR inhibitors,rapamycin(100 nmol/L)and KU-0063794(5mmol/L),could inhibit mTOR activity in KFs by western blotting.Then we explored the reverse effects on autophagy activity induced by mTOR inhibitors in the presence of lysosomal protease inhibitors by western blotting.Results:It demonstrated elevated expression of mTOR,S6,and their activated forms in KTs,and an elevated expression of p-S6 Ser235/236 in KFs,suggesting mTOR was activated in keloid.Less LC3 and Beclin1 were expressed in the cytoplasm of KFs,whereas Ubiquitin was abundantly expressed in KTs compared with extra-lesional tissues.In addition,at the cellular level,an impeded conversion of LC3-I to LC3-II was shown in KFs and the formation of acid vesicles were also decreased in KFs compared with normal fibroblasts(NFs),indicating that autophagy activity is defective in keloid.mTOR inhibitors,Rapamycin(E-64d+pepstatin vs.rapamycin+E-64d+pepstatin:[0.88±0.35]vs.[1.56±0.46],F=5.56,P=0.049)and KU-0063794(E-64d+pepstatin vs.KU-0063794+E-64d+pepstatin:[0.92±0.22]vs.[1.51±0.25],F=25.88,P=0.011)can reverse the inhibition effect on autophagy of KFs while inhibiting mTOR activity.Conclusion:Autophagy machinery is inhibited in keloid which is regulated by mTOR-dependent pathway.
基金This work was supported by the CAMS Innovation Fund for Medical Sciences(No.CIFMS-2017-I2M-1-017)Nanjing Incubation Program for National Clinical Research Center(No.2019060001)。
文摘Objective:Blocking mechanistic target of rapamycin(mTOR)activation with mTOR inhibitors has promising therapeutic potential for keloids.However,the precise mechanism of mTOR inhibitors remains unclear.This study was aimed to investigate the role of autophagy machinery in the anti-keloid effects of mTOR inhibitors.Methods:We first validated the biological effects induced by the mTOR inhibitors rapamycin(100 nmol/L)and KU-0063794(5μmol/L)on the proliferation,apoptosis,migration,and collagen synthesis of keloid fibroblasts(KFs)derived from Han Chinese persons through a Cell Counting Kit-8 assay,5-Bromo-2’-deoxyuridine incorporation,Annexin V/propidium iodide staining,migration,and western blotting.To explore whether autophagy machinery is involved in the anti-keloid effects of mTOR inhibitors,we first blocked the autophagy activation induced by rapamycin and KU-0063794 with a pharmacological autophagy inhibitor(wortmannin)or by silencing the key autophagy gene(ATG5),and we then re-evaluated these biological effects on KFs.Results:Blocking mTOR activation with either rapamycin or KU-0063794 completely inhibited proliferation,migration,and collagen synthesis of primary KFs but did not affect apoptosis.Incubating KFs with the autophagy inhibitor wortmannin or performingATG5 silencing abrogated the subsequent activation of autophagic activity induced by rapamycin(rapamycin+E-64d+pepstatinvs.rapamycin+wortmannin+E-64d+pepstatin:1.88±0.38vs.1.02±0.35,F=6.86,P=0.013),(non-sense control+rapamycinvs.ATG5 small interfering RNA+rapamycin:1.46±0.18vs.0.75±0.20,respectively;F=7.68,P=0.01)or KU-0063794(KU-0063794+E-64d+pepstatinvs.KU-0063794+wortmannin+E-64d+pepstatin:1.65±0.35vs.0.76±0.17,F=10.01,P=0.004),(NC+KU-0063794vs.ATG5 small interfering RNA+KU-0063794:1.59±0.50vs.0.77±0.09,F=5.93,P=0.02)as evidenced by decreased accumulation of LC3-II.However,blockage of autophagy induction in mTOR inhibitor-treated KFs with both methods did not disturb their anti-keloid effects,such as inhibition of cell viability,cell migration,and collagen synthesis(P>0.05 each).Conclusion:Blocking mTOR activation with the mTOR inhibitors rapamycin and KU-0063794 showed anti-keloid effects in KFs.Restoration of autophagy inhibition by mTOR inhibitors does not contribute to their anti-keloid effects.
