Liquid chromatography tandem mass spectrometry(LC-MS/MS)has gradually become a promising alternative to ligand binding assay for the bioanalysis of biotherapeutic molecules,due to its rapid method development and high...Liquid chromatography tandem mass spectrometry(LC-MS/MS)has gradually become a promising alternative to ligand binding assay for the bioanalysis of biotherapeutic molecules,due to its rapid method development and high accuracy.In this study,we established a new LC-MS/MS method for the determination of the anti-sclerostin monoclonal antibody(SHR-1222)in cynomolgus monkey serum,and compared it to a previous electrochemiluminescence method.The antibody was quantified by detecting the surrogate peptide obtained by trypsin digestion.The surrogate peptide was carefully selected by investigating its uniqueness,stability and MS response.The quantitative range of the proposed method was 2.00-500μg/mL,and this verified method was successfully applied to the toxicokinetic assessment of SHR-1222 in cynomolgus monkey serum.It was found that the concentrations of SHR-1222 in cynomolgus monkeys displayed an excellent agreement between the LC-MS/MS and electrochemiluminescence methods(ratios of drug exposure,0.8-1.0).Notably,two monkeys in the60 mg/kg dose group had abnormal profiles with a low detection value of SHR-1222 in their individual sample.Combining the high-level anti-drug antibodies(ADAs)in these samples and the consistent quantitative results of the two methods,we found that the decreased concentration of SHR-1222 was due to the accelerated clearance mediated by ADAs rather than the interference of ADAs to the detection platform.Taken together,we successfully developed an accurate,efficient and cost-effective LC-MS/MS method for the quantification of SHR-1222 in serum samples,which could serve as a powerful tool to improve the preclinical development of antibody drugs.展开更多
In order to explore the stability of a liquid crystal(LC)system doped withγ-Fe_(2)O_(3)nanoparticles,the physical properties(clearing point,dielectric properties),electro-optical properties and residual direct-curren...In order to explore the stability of a liquid crystal(LC)system doped withγ-Fe_(2)O_(3)nanoparticles,the physical properties(clearing point,dielectric properties),electro-optical properties and residual direct-current voltage(RDCV)of the doped LC system were measured and evaluated at different times.First,the temperature was controlled by precision hot stage,and the clearing point temperature of doped LC was observed and measured by a polarized optical microscope.Using a precision LCR meter,we measured the capacitance-voltage curves of the doped LC system at the temperature of 27℃.The dielectric constant of doped LC was calculated by the dualcell capacitance method.Then,the electro-optical properties of the doped LC system were measured.Finally,the RDCV of the doped LC system was measured and calculated.After five months,the parameters of the doped LC system were re-measured and analyzed under the same conditions to evaluate its stability.The experimental results show that,within five months,the clearing point change rate of doped LC is in the range of 0.24%-1.37%,the change of dielectric anisotropy is in the range of 0.035-0.2,the curves of electro-optical properties are basically fitted,and the change rate of saturated RDCV is about 11.2%,which basically indicate that the LC system doped withγ-Fe_(2)O_(3)nanoparticles has good stability.展开更多
Objective:The aim of the study was to observe the expression of Bcl-2 and its phosphorylation in Molt-4 cells induced by tumor necrosis factor-α(TNF-α),and to investigate the possible mechanism of cell cycle specifi...Objective:The aim of the study was to observe the expression of Bcl-2 and its phosphorylation in Molt-4 cells induced by tumor necrosis factor-α(TNF-α),and to investigate the possible mechanism of cell cycle specificity of apoptosis.Methods:Exponentially growing Molt-4 cells were treated with TNF-α.Apoptosis was detected by DNA fragmentation assay.API method was applied to illustrate the cell cycle specificity of apoptotic cells.Cells of sub-phases were sorted by FACSvantage flow cytometer and then submitted to immunoblot.Results:Molt-4 cells which were treated with TNF-α went to apoptosis and showed a DNA ladder pattern.Most apoptosis happened in G1-phase of cell cycle.Bcl-2 expression increased for the Molt-4 cells treated with TNF-α.The phosphorylation state of Bcl-2 was only presented in G1-phase cells,in accordance with the specified time and cell cycle phase of apoptosis.Conclusion:The phosphorylation of Bcl-2 in the Molt-4 cells treated with TNF-α happened with the same cell cycle specificity as cell apoptosis.The cell cycle specificity of Bcl-2 phosphorylation was one of the mechanisms of receptor-mediated apoptosis.The cell cycle machine can trigger the apoptosis program.展开更多
In this article,a grinding force model,which is on the basis of cutting process of single abrasive grains combined with the method of theoretical derivation and empirical formula by analyzing the formation mechanism o...In this article,a grinding force model,which is on the basis of cutting process of single abrasive grains combined with the method of theoretical derivation and empirical formula by analyzing the formation mechanism of grinding force,was established.Three key factors have been taken into accounts in this model,such as the contact friction force between abrasive grains and materials,the plastic deformation of material in the process of abrasive plowing,and the shear strain effect of material during the process of cutting chips formation.The model was finally validated by the orthogonal grinding experiment of powder metallurgy nickel-based superalloy FGH96 by using the electroplated CBN abrasive wheel.Grinding force values of prediction and experiment were in good consistency.The errors of tangential grinding force and normal grinding force were 9.8%and 13.6%,respectively.The contributions of sliding force,plowing force and chip formation force were also analyzed.In addition,the tangential forces of sliding,plowing and chip formation are 14%,19%and 11%of the normal forces on average,respectively.The pro-posed grinding forcemodel is not only in favor of optimizing the grinding parameters and improving grinding efficiency,but also contributes to study some other grinding subjects(e.g.abrasive wheel wear,grinding heat,residual stress).展开更多
Creep feed profile grinding of the fir-tree blade root forms of single crystal nickel-based superalloy was conducted using microcrystalline alumina abrasive wheels in the present study. The grinding force and the surf...Creep feed profile grinding of the fir-tree blade root forms of single crystal nickel-based superalloy was conducted using microcrystalline alumina abrasive wheels in the present study. The grinding force and the surface quality in terms of surface topography, subsurface microstructure,microhardness and residual stress obtained under different grinding conditions were evaluated comparatively. Experimental results indicated that the grinding force was influenced significantly by the competing predominance between the grinding parameters and the cross-sectional root workpiece profile. In addition, the root workpiece surface, including the root peak and valley regions, was produced with the large difference in surface quality due to the nonuniform grinding loads along the root workpiece profile in normal section. Detailed results showed that the surface roughness, subsurface plastic deformation and work hardening level of the root valley region were higher by up to25%, 20% and 7% in average than those obtained in the root peak region, respectively, in the current investigation. Finally, the superior parameters were recommended in the creep feed profile grinding of the fir-tree blade root forms. This study is helpful to provide industry guidance to optimize the machining process for the high-valued parts with complicated profiles.展开更多
Single-pass honing is an important machining method for finish machining of holes,which can meet the requirement for high efficiency and consistency of holes.Characterization and life prediction of single-pass honing ...Single-pass honing is an important machining method for finish machining of holes,which can meet the requirement for high efficiency and consistency of holes.Characterization and life prediction of single-pass honing tool are necessary to improve the machining accuracy of holes honed,especially dimension accuracy.Single-pass honing tool is a single layer abrasive tool with fixed dimension,which still remains problematic for characterization and life prediction.For fuel injection nozzles with bore diameter under 1 mm,the stiffness of the single-pass honing tool is poor.This article presents a novel analytical model that predicts life of the tool with poor stiffness.Firstly,according to the bore diameter and dimension tolerance,the single-pass honing tool is designed and manufactured.Based on the prepared single-pass honing tool,the measurement and characterization methods are established.Furthermore,the tool wear tests are carried out,and the tool contour evolution model is established to predict the tool life.展开更多
基金supported by the National Natural Science Foundation of China(Grant No.81521005)the National Key Research Project of the Chinese Academy of Sciences(Grant No.XDA12050306)。
文摘Liquid chromatography tandem mass spectrometry(LC-MS/MS)has gradually become a promising alternative to ligand binding assay for the bioanalysis of biotherapeutic molecules,due to its rapid method development and high accuracy.In this study,we established a new LC-MS/MS method for the determination of the anti-sclerostin monoclonal antibody(SHR-1222)in cynomolgus monkey serum,and compared it to a previous electrochemiluminescence method.The antibody was quantified by detecting the surrogate peptide obtained by trypsin digestion.The surrogate peptide was carefully selected by investigating its uniqueness,stability and MS response.The quantitative range of the proposed method was 2.00-500μg/mL,and this verified method was successfully applied to the toxicokinetic assessment of SHR-1222 in cynomolgus monkey serum.It was found that the concentrations of SHR-1222 in cynomolgus monkeys displayed an excellent agreement between the LC-MS/MS and electrochemiluminescence methods(ratios of drug exposure,0.8-1.0).Notably,two monkeys in the60 mg/kg dose group had abnormal profiles with a low detection value of SHR-1222 in their individual sample.Combining the high-level anti-drug antibodies(ADAs)in these samples and the consistent quantitative results of the two methods,we found that the decreased concentration of SHR-1222 was due to the accelerated clearance mediated by ADAs rather than the interference of ADAs to the detection platform.Taken together,we successfully developed an accurate,efficient and cost-effective LC-MS/MS method for the quantification of SHR-1222 in serum samples,which could serve as a powerful tool to improve the preclinical development of antibody drugs.
基金Project supported by the Natural Science Foundation of Hebei Province of China (Grant Nos. A2019202235 and A2017202004)Research and Practice Project of Hebei Provincial Higher Education and Teaching Reform (Grant No. 2017GJJG018)+2 种基金Research Projects of Undergraduate Education and Teaching Reform in Hebei University of Technology (Grant No. 201802003)the Key Subject Construction Project of Hebei Provincial Universitiesthe Special Project of China Association of Higher Education (Grant No. 21LKYB05)
文摘In order to explore the stability of a liquid crystal(LC)system doped withγ-Fe_(2)O_(3)nanoparticles,the physical properties(clearing point,dielectric properties),electro-optical properties and residual direct-current voltage(RDCV)of the doped LC system were measured and evaluated at different times.First,the temperature was controlled by precision hot stage,and the clearing point temperature of doped LC was observed and measured by a polarized optical microscope.Using a precision LCR meter,we measured the capacitance-voltage curves of the doped LC system at the temperature of 27℃.The dielectric constant of doped LC was calculated by the dualcell capacitance method.Then,the electro-optical properties of the doped LC system were measured.Finally,the RDCV of the doped LC system was measured and calculated.After five months,the parameters of the doped LC system were re-measured and analyzed under the same conditions to evaluate its stability.The experimental results show that,within five months,the clearing point change rate of doped LC is in the range of 0.24%-1.37%,the change of dielectric anisotropy is in the range of 0.035-0.2,the curves of electro-optical properties are basically fitted,and the change rate of saturated RDCV is about 11.2%,which basically indicate that the LC system doped withγ-Fe_(2)O_(3)nanoparticles has good stability.
基金Supported by grants from the Clinical Key Subject Foundation of Health Ministry of China (No.20012537)National Science and Technology Department "973" Tumor Project (No.2004CB518705)
文摘Objective:The aim of the study was to observe the expression of Bcl-2 and its phosphorylation in Molt-4 cells induced by tumor necrosis factor-α(TNF-α),and to investigate the possible mechanism of cell cycle specificity of apoptosis.Methods:Exponentially growing Molt-4 cells were treated with TNF-α.Apoptosis was detected by DNA fragmentation assay.API method was applied to illustrate the cell cycle specificity of apoptotic cells.Cells of sub-phases were sorted by FACSvantage flow cytometer and then submitted to immunoblot.Results:Molt-4 cells which were treated with TNF-α went to apoptosis and showed a DNA ladder pattern.Most apoptosis happened in G1-phase of cell cycle.Bcl-2 expression increased for the Molt-4 cells treated with TNF-α.The phosphorylation state of Bcl-2 was only presented in G1-phase cells,in accordance with the specified time and cell cycle phase of apoptosis.Conclusion:The phosphorylation of Bcl-2 in the Molt-4 cells treated with TNF-α happened with the same cell cycle specificity as cell apoptosis.The cell cycle specificity of Bcl-2 phosphorylation was one of the mechanisms of receptor-mediated apoptosis.The cell cycle machine can trigger the apoptosis program.
基金financial support for this work by the National Natural Science Foundation of China(Nos.51775275,51921003 and 51905363)the Funding for Outstanding Doctoral Dissertation in NUAA of China(No.BCXJ19-06)+1 种基金the Natural Science Foundation of Jiangsu Province of China(No.BK20190940)the Natural Science Foundation of the Jiangsu Higher Education Institutions of China(No.19KJB460008)。
文摘In this article,a grinding force model,which is on the basis of cutting process of single abrasive grains combined with the method of theoretical derivation and empirical formula by analyzing the formation mechanism of grinding force,was established.Three key factors have been taken into accounts in this model,such as the contact friction force between abrasive grains and materials,the plastic deformation of material in the process of abrasive plowing,and the shear strain effect of material during the process of cutting chips formation.The model was finally validated by the orthogonal grinding experiment of powder metallurgy nickel-based superalloy FGH96 by using the electroplated CBN abrasive wheel.Grinding force values of prediction and experiment were in good consistency.The errors of tangential grinding force and normal grinding force were 9.8%and 13.6%,respectively.The contributions of sliding force,plowing force and chip formation force were also analyzed.In addition,the tangential forces of sliding,plowing and chip formation are 14%,19%and 11%of the normal forces on average,respectively.The pro-posed grinding forcemodel is not only in favor of optimizing the grinding parameters and improving grinding efficiency,but also contributes to study some other grinding subjects(e.g.abrasive wheel wear,grinding heat,residual stress).
基金financial support for this work by the National Natural Science Foundation of China (No. 51775275)the Funding of Jiangsu Innovation Program for Graduate Education of China (KYCX170245)+2 种基金the Funding for Outstanding Doctoral Dissertation in NUAA of China (BCXJ17-04)the Fundamental Research Funds for the Central University of China (No. NP2018110)the National Science and Technology Major Project and the Six Talents Summit Project in Jiangsu Province of China (No.JXQC-002)。
文摘Creep feed profile grinding of the fir-tree blade root forms of single crystal nickel-based superalloy was conducted using microcrystalline alumina abrasive wheels in the present study. The grinding force and the surface quality in terms of surface topography, subsurface microstructure,microhardness and residual stress obtained under different grinding conditions were evaluated comparatively. Experimental results indicated that the grinding force was influenced significantly by the competing predominance between the grinding parameters and the cross-sectional root workpiece profile. In addition, the root workpiece surface, including the root peak and valley regions, was produced with the large difference in surface quality due to the nonuniform grinding loads along the root workpiece profile in normal section. Detailed results showed that the surface roughness, subsurface plastic deformation and work hardening level of the root valley region were higher by up to25%, 20% and 7% in average than those obtained in the root peak region, respectively, in the current investigation. Finally, the superior parameters were recommended in the creep feed profile grinding of the fir-tree blade root forms. This study is helpful to provide industry guidance to optimize the machining process for the high-valued parts with complicated profiles.
基金the National Science and Technology Major Project of China(2017-Ⅶ-0015-0111)。
文摘Single-pass honing is an important machining method for finish machining of holes,which can meet the requirement for high efficiency and consistency of holes.Characterization and life prediction of single-pass honing tool are necessary to improve the machining accuracy of holes honed,especially dimension accuracy.Single-pass honing tool is a single layer abrasive tool with fixed dimension,which still remains problematic for characterization and life prediction.For fuel injection nozzles with bore diameter under 1 mm,the stiffness of the single-pass honing tool is poor.This article presents a novel analytical model that predicts life of the tool with poor stiffness.Firstly,according to the bore diameter and dimension tolerance,the single-pass honing tool is designed and manufactured.Based on the prepared single-pass honing tool,the measurement and characterization methods are established.Furthermore,the tool wear tests are carried out,and the tool contour evolution model is established to predict the tool life.
