Objective Acute myeloid leukemia(AML)is a highly heterogeneous disease,and molecular events such as DNMT3A gene mutations are associated with poor prognosis in AML patients.Consequently,there is an urgent need for a n...Objective Acute myeloid leukemia(AML)is a highly heterogeneous disease,and molecular events such as DNMT3A gene mutations are associated with poor prognosis in AML patients.Consequently,there is an urgent need for a novel therapeutic approach for AML.Methods DNMT3A mRNA and protein expression were confirmed in DNMT3A-mutant AML cells via RT-qPCR and Western blotting.Cell proliferation and apoptosis were assessed via CCK-8 and Annexin V/PI staining,respectively.Flow cytometry was used to analyze surface antigens and CD44v6 CAR-T-cell transfection efficiency.CD44v6-directed CAR plasmids were constructed,and lentiviruses were packaged.Methylation-specific PCR was used to evaluate differences in promoter methylation,whereas ELISA was used to measure cytokine secretion.Results In this study,we found that the DNMT3A-mutant group presented significantly increased expression of CD44v6 on the cell surface.Methylation of the CD44 promoter region was lower in the mutant group than in the control group.CD44v6 CAR-T cells exhibited specific cytotoxicity against DNMT3A-mutant AML cells.Furthermore,pretreatment with low concentrations of decitabine significantly enhanced the killing effect of CD44v6 CAR-T cells on DNMT3A-mutant AML cells(P<0.05).Additionally,decitabine treatment upregulated the expression of CD44v6 on the surface of DNMT3A-mutant AML cells(P<0.05).Conclusion CD44v6 is a promising CAR-T-cell therapy target in AML patients with DNMT3A mutations.Notably,treatment with decitabine resulted in increased CD44v6 expression on the cell surface of DNMT3A-mutant AML cells.This increase in CD44v6 expression facilitates improved recognition and targeting by CD44v6 CAR-T cells.展开更多
This study focuses on testing and quality measurement and analysis of VoIPv6 performance. A client, server codes were developed using FreeBSD. This is a step before analyzing the Architectures of VoIPv6 in the current...This study focuses on testing and quality measurement and analysis of VoIPv6 performance. A client, server codes were developed using FreeBSD. This is a step before analyzing the Architectures of VoIPv6 in the current internet in order for it to cope with IPv6 traffic transmission requirements in general and specifically voice traffic, which is being attracting the efforts of research, bodes currently. These tests were conducted in the application level without looking into the network level of the network. VoIPv6 performance tests were conducted in the current tunneled and native IPv6 aiming for better end-to-end VoIPv6 performance. The results obtained in this study were shown in deferent codec's for different bit rates in Kilo bits per second, which act as an indicator for the better performance of G.711 compared with the rest of the tested codes.展开更多
基金supported by the National Natural Science Foundation of China(No.82270177)Chen Xiaoping Foundation for the Development of Science and Technology of Hubei Province(CXPJJH122001-2221).
文摘Objective Acute myeloid leukemia(AML)is a highly heterogeneous disease,and molecular events such as DNMT3A gene mutations are associated with poor prognosis in AML patients.Consequently,there is an urgent need for a novel therapeutic approach for AML.Methods DNMT3A mRNA and protein expression were confirmed in DNMT3A-mutant AML cells via RT-qPCR and Western blotting.Cell proliferation and apoptosis were assessed via CCK-8 and Annexin V/PI staining,respectively.Flow cytometry was used to analyze surface antigens and CD44v6 CAR-T-cell transfection efficiency.CD44v6-directed CAR plasmids were constructed,and lentiviruses were packaged.Methylation-specific PCR was used to evaluate differences in promoter methylation,whereas ELISA was used to measure cytokine secretion.Results In this study,we found that the DNMT3A-mutant group presented significantly increased expression of CD44v6 on the cell surface.Methylation of the CD44 promoter region was lower in the mutant group than in the control group.CD44v6 CAR-T cells exhibited specific cytotoxicity against DNMT3A-mutant AML cells.Furthermore,pretreatment with low concentrations of decitabine significantly enhanced the killing effect of CD44v6 CAR-T cells on DNMT3A-mutant AML cells(P<0.05).Additionally,decitabine treatment upregulated the expression of CD44v6 on the surface of DNMT3A-mutant AML cells(P<0.05).Conclusion CD44v6 is a promising CAR-T-cell therapy target in AML patients with DNMT3A mutations.Notably,treatment with decitabine resulted in increased CD44v6 expression on the cell surface of DNMT3A-mutant AML cells.This increase in CD44v6 expression facilitates improved recognition and targeting by CD44v6 CAR-T cells.
文摘This study focuses on testing and quality measurement and analysis of VoIPv6 performance. A client, server codes were developed using FreeBSD. This is a step before analyzing the Architectures of VoIPv6 in the current internet in order for it to cope with IPv6 traffic transmission requirements in general and specifically voice traffic, which is being attracting the efforts of research, bodes currently. These tests were conducted in the application level without looking into the network level of the network. VoIPv6 performance tests were conducted in the current tunneled and native IPv6 aiming for better end-to-end VoIPv6 performance. The results obtained in this study were shown in deferent codec's for different bit rates in Kilo bits per second, which act as an indicator for the better performance of G.711 compared with the rest of the tested codes.
