The Chinese Pharmacopoeia 2020 edition was reviewed and approved by the National Medical Products Administration and the National Health Commission of the People’s Republic of China in July 2020.The current edition w...The Chinese Pharmacopoeia 2020 edition was reviewed and approved by the National Medical Products Administration and the National Health Commission of the People’s Republic of China in July 2020.The current edition was officially implemented on December 30,2020.The general chapters of the Chinese Pharmacopoeia discuss the general testing methods and guidelines,which are the common requirements and basis for the implementation of drug standards in the Chinese Pharmacopoeia.Owing to adherence to the principles of scientificity,versatility,operability,and sustainable development,there is an improvement in the general chapters of the 2020 edition over those of the previous editions.Further,the application of advanced and mature analytical techniques has expanded,the development of testing methods for exogenous pollutants in traditional Chinese medicines has been strengthened,and technical requirements are now better harmonized with international standards.The updated edition provides technical and methodological support to ensure safety,effectiveness,and control of pharmaceuticals in China and will play an important and active role in encouraging the application of advanced technologies,improving the quality control of medicines,and strengthening the means of drug regulation in China.This review provides a comprehensive introduction of the main features of and changes to the general chapters in the Chinese Pharmacopoeia 2020 edition and aims to provide reference for its correct understanding and accurate implementation.展开更多
Objective:Death receptor 4(DR4;TRAIL-R1)critically mediates extrinsic apoptosis cascades via binding to TNF-related apoptosis-inducing ligand(TRAIL).However,intrinsic and/or acquired resistance are observed in the cli...Objective:Death receptor 4(DR4;TRAIL-R1)critically mediates extrinsic apoptosis cascades via binding to TNF-related apoptosis-inducing ligand(TRAIL).However,intrinsic and/or acquired resistance are observed in the clinical application of TRAIL.The aim of this study was to investigate the function and molecular mechanism of CD13 in the TRAIL/DR4 pathway against tumor cells,and provide a new strategy for improving therapeutic efficacy or overcoming TRAIL-resistance.Methods:TRAIL protein was expressed as a secretory protein in a Pichia pastoris expression system and was isolated and purified by affinity chromatography.The cell viability and apoptosis were evaluated with MTT(thiazolyl blue tetrazolium bromide)assays and annexin V-FITC/PI staining with flow cytometry analysis,respectively.Western blot analysis was used to detect the levels of the indicated proteins in tumor cells.DR4 degradation or stability was examined with cycloheximide chase assays,and cell surface DR4 was assessed with flow cytometric analysis after staining with a FITC-conjugated antibody.The effects of cell migration were determined with Transwell and gelatin zymography assays.A xenograft nude mouse model was used to detect the anti-tumor effect in vivo,and the proliferation in tumor tissues was examined with immunohistochemical staining.Results:CD13 inhibition potently sensitized tumor cells to TRAIL-induced killing,including proliferation inhibition,increased apoptosis,and migration suppression.In addition,the inhibition of CD13 elevated both total cellular expression and cell surface DR4 through stabilizing DR4 by suppressing its degradation.DR4 si RNA attenuated the enhanced anti-tumor effects of TRAIL plus CD13 inhibition.Interestingly,these phenomena were p-ERK1/2 independent,although p-ERK1/2 down-regulation was tightly correlated with the cooperation of TRAIL and CD13 inhibition.Moreover,a synergistic decrease in tumor growth was surprisingly achieved in the xenograft model by treatment of TRAIL with a CD13 inhibitor(**P<0.01,CDI=0.47).Conclusions:CD13 inhibition cooperates with TRAIL in enhancing DR4-mediated cell death,through the up-regulation and stabilization of DR4 in a p-ERK1/2-independent manner.Thus CD13 inhibition has emerged as an effective strategy for TRAIL/DR4-based therapy.展开更多
Background:Azithromycin is a member of macrolide antibiotics,and has been reported to inhibit the proliferation of cancer cells.However,the underlying mechanisms are not been fully elucidated.Tumor necrosis factor-rel...Background:Azithromycin is a member of macrolide antibiotics,and has been reported to inhibit the proliferation of cancer cells.However,the underlying mechanisms are not been fully elucidated.Tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)selectively targets tumor cells without damaging healthy cells.In the present study,we examined whether azithromycin is synergistic with TRAIL,and if so,the underlying mechanisms in colon cancers.Methods:HCT-116,SW480,SW620 and DiFi cells were treated with azithromycin,purified TRAIL,or their combina-tion.A sulforhoddamine B assay was used to examine cell survival.Apoptosis was examined using annexin V-FITC/PI staining,and autophagy was observed by acridine orange staining.Western blot analysis was used to detect protein expression levels.In mechanistic experiments,siRNAs were used to knockdown death receptors(DR4,DR5)and LC-3B.The anticancer effect of azithromycin and TRAIL was also examined in BALB/c nude mice carrying HCT-116 xenografts.Results:Azithromycin decreased the proliferation of HCT-116 and SW480 cells in a dose-dependent manner.Combination of azithromycin and TRAIL inhibited tumor growth in a manner that could not be explained by additive effects.Azithromycin increased the expressions of DR4,DR5,p62 and LC-3B proteins and potentiated induction of apoptosis by TRAIL.Knockdown of DR4 and DR5 with siRNAs increased cell survival rate and decreased the expression of cleaved-PARP induced by the combination of azithromycin and TRAIL.LC-3B siRNA and CQ potentiated the anti-proliferation activity of TRAIL alone,and increased the expressions of DR4 and DR5.Conclusion:The synergistic antitumor effect of azithromycin and TRAIL mainly relies on the up-regulations of DR4 and DR5,which in turn result from LC-3B-involved autophagy inhibition.展开更多
基金financial support from the Chinese Pharmacopoeia Commission Drug Standard Promoting Funds and Comprehensive Reform of the Chinese Drug and Medical Device Review and Approval System Funds(2015e2020)。
文摘The Chinese Pharmacopoeia 2020 edition was reviewed and approved by the National Medical Products Administration and the National Health Commission of the People’s Republic of China in July 2020.The current edition was officially implemented on December 30,2020.The general chapters of the Chinese Pharmacopoeia discuss the general testing methods and guidelines,which are the common requirements and basis for the implementation of drug standards in the Chinese Pharmacopoeia.Owing to adherence to the principles of scientificity,versatility,operability,and sustainable development,there is an improvement in the general chapters of the 2020 edition over those of the previous editions.Further,the application of advanced and mature analytical techniques has expanded,the development of testing methods for exogenous pollutants in traditional Chinese medicines has been strengthened,and technical requirements are now better harmonized with international standards.The updated edition provides technical and methodological support to ensure safety,effectiveness,and control of pharmaceuticals in China and will play an important and active role in encouraging the application of advanced technologies,improving the quality control of medicines,and strengthening the means of drug regulation in China.This review provides a comprehensive introduction of the main features of and changes to the general chapters in the Chinese Pharmacopoeia 2020 edition and aims to provide reference for its correct understanding and accurate implementation.
基金supported by grants from the Beijing Natural Science Foundation(Grant No.7202132)CAMS Innovation Fund for Medical Sciences(CIFMS+1 种基金Grant No.2016-I2M-02-002)“Significant New Drug Development”Major Science and Technology Development Projects of China(Grant No.2018ZX09711001-007-002)。
文摘Objective:Death receptor 4(DR4;TRAIL-R1)critically mediates extrinsic apoptosis cascades via binding to TNF-related apoptosis-inducing ligand(TRAIL).However,intrinsic and/or acquired resistance are observed in the clinical application of TRAIL.The aim of this study was to investigate the function and molecular mechanism of CD13 in the TRAIL/DR4 pathway against tumor cells,and provide a new strategy for improving therapeutic efficacy or overcoming TRAIL-resistance.Methods:TRAIL protein was expressed as a secretory protein in a Pichia pastoris expression system and was isolated and purified by affinity chromatography.The cell viability and apoptosis were evaluated with MTT(thiazolyl blue tetrazolium bromide)assays and annexin V-FITC/PI staining with flow cytometry analysis,respectively.Western blot analysis was used to detect the levels of the indicated proteins in tumor cells.DR4 degradation or stability was examined with cycloheximide chase assays,and cell surface DR4 was assessed with flow cytometric analysis after staining with a FITC-conjugated antibody.The effects of cell migration were determined with Transwell and gelatin zymography assays.A xenograft nude mouse model was used to detect the anti-tumor effect in vivo,and the proliferation in tumor tissues was examined with immunohistochemical staining.Results:CD13 inhibition potently sensitized tumor cells to TRAIL-induced killing,including proliferation inhibition,increased apoptosis,and migration suppression.In addition,the inhibition of CD13 elevated both total cellular expression and cell surface DR4 through stabilizing DR4 by suppressing its degradation.DR4 si RNA attenuated the enhanced anti-tumor effects of TRAIL plus CD13 inhibition.Interestingly,these phenomena were p-ERK1/2 independent,although p-ERK1/2 down-regulation was tightly correlated with the cooperation of TRAIL and CD13 inhibition.Moreover,a synergistic decrease in tumor growth was surprisingly achieved in the xenograft model by treatment of TRAIL with a CD13 inhibitor(**P<0.01,CDI=0.47).Conclusions:CD13 inhibition cooperates with TRAIL in enhancing DR4-mediated cell death,through the up-regulation and stabilization of DR4 in a p-ERK1/2-independent manner.Thus CD13 inhibition has emerged as an effective strategy for TRAIL/DR4-based therapy.
基金supported by grants from the National Natural Science Foundation of China(81373437 and 81621064)CAMS Innovation Fund for Medical Sciences(CIFMS)(2016-I2M-02-002).
文摘Background:Azithromycin is a member of macrolide antibiotics,and has been reported to inhibit the proliferation of cancer cells.However,the underlying mechanisms are not been fully elucidated.Tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)selectively targets tumor cells without damaging healthy cells.In the present study,we examined whether azithromycin is synergistic with TRAIL,and if so,the underlying mechanisms in colon cancers.Methods:HCT-116,SW480,SW620 and DiFi cells were treated with azithromycin,purified TRAIL,or their combina-tion.A sulforhoddamine B assay was used to examine cell survival.Apoptosis was examined using annexin V-FITC/PI staining,and autophagy was observed by acridine orange staining.Western blot analysis was used to detect protein expression levels.In mechanistic experiments,siRNAs were used to knockdown death receptors(DR4,DR5)and LC-3B.The anticancer effect of azithromycin and TRAIL was also examined in BALB/c nude mice carrying HCT-116 xenografts.Results:Azithromycin decreased the proliferation of HCT-116 and SW480 cells in a dose-dependent manner.Combination of azithromycin and TRAIL inhibited tumor growth in a manner that could not be explained by additive effects.Azithromycin increased the expressions of DR4,DR5,p62 and LC-3B proteins and potentiated induction of apoptosis by TRAIL.Knockdown of DR4 and DR5 with siRNAs increased cell survival rate and decreased the expression of cleaved-PARP induced by the combination of azithromycin and TRAIL.LC-3B siRNA and CQ potentiated the anti-proliferation activity of TRAIL alone,and increased the expressions of DR4 and DR5.Conclusion:The synergistic antitumor effect of azithromycin and TRAIL mainly relies on the up-regulations of DR4 and DR5,which in turn result from LC-3B-involved autophagy inhibition.
