Hepatitis B virus(HBV)is a serious global public health concern.Although nucleoside drugs and interferons can significantly inhibit HBV replication,issues such as drug resistance and low clinical cure rates remain.Tra...Hepatitis B virus(HBV)is a serious global public health concern.Although nucleoside drugs and interferons can significantly inhibit HBV replication,issues such as drug resistance and low clinical cure rates remain.Traditional Chinese medicine(TCM)is widely used in the treatment of chronic hepatitis B(CHB)in China,with anti-inflammatory,anti-fibrotic,and liver-protective effects;however,reports on its antiviral effects are still inconsistent.We retrieved multicenter clinical studies and meta-analyses of TCM treatment for CHB over the past two decades.The results revealed that TCM has a certain anti-HBV effect,and when combined with antiviral drugs,it can significantly improve antiviral efficacy.It was demonstrated that TCM most effectively promotes serum HBV e antigen conversion to negative,followed by the ability to reduce HBV DNA levels,facilitating HBV surface antigen loss,and improving the treatment of CHB.展开更多
Histone modification including H3 lysine 79 methylation (H3K79me) plays a key role during gene transcription and DNA damage repair. DOT1L, the sole methyltransferase for three states of H3K79me, is implicated in leuke...Histone modification including H3 lysine 79 methylation (H3K79me) plays a key role during gene transcription and DNA damage repair. DOT1L, the sole methyltransferase for three states of H3K79me, is implicated in leukemia, colorectal cancer, and dilated cardiomyopathy. However, understanding of DOT1L and H3K79me in these pathways and disease pathogenesis has been limited due to the difficulty of working with DOT1L protein. For instance, locus-specific or genome-wide binding sites of DOT1L revealed by chromatin immunoprecipitation (ChIP)-based methods are necessary for inferring its functions, but high-quality ChIP-grade antibodies are currently not available. Herein we have developed a knock-in approach to tag endogenous DOT1L with 3 × Flag at its C-terminal domain to follow functional analyses. The knock-in was facilitated by using TALENs to induce a targeted double-strand break at the endogenous DOTIL to stimulate local homologous recombination at that site. The single cell colonies with successful knock-in were isolated and verified by different methods. We also demonstrated that tagged DOT1L maintains its normal function in terms of methylation and that the engineered cells would be very useful for further studies.展开更多
基金Supported by The Funding of Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences,No.CI2021A00810The Escort Project of Guang’anmen Hosital,China Academy of Chinese Medical Science-Backbone Talent Cultivation Project,No.9323013。
文摘Hepatitis B virus(HBV)is a serious global public health concern.Although nucleoside drugs and interferons can significantly inhibit HBV replication,issues such as drug resistance and low clinical cure rates remain.Traditional Chinese medicine(TCM)is widely used in the treatment of chronic hepatitis B(CHB)in China,with anti-inflammatory,anti-fibrotic,and liver-protective effects;however,reports on its antiviral effects are still inconsistent.We retrieved multicenter clinical studies and meta-analyses of TCM treatment for CHB over the past two decades.The results revealed that TCM has a certain anti-HBV effect,and when combined with antiviral drugs,it can significantly improve antiviral efficacy.It was demonstrated that TCM most effectively promotes serum HBV e antigen conversion to negative,followed by the ability to reduce HBV DNA levels,facilitating HBV surface antigen loss,and improving the treatment of CHB.
文摘Histone modification including H3 lysine 79 methylation (H3K79me) plays a key role during gene transcription and DNA damage repair. DOT1L, the sole methyltransferase for three states of H3K79me, is implicated in leukemia, colorectal cancer, and dilated cardiomyopathy. However, understanding of DOT1L and H3K79me in these pathways and disease pathogenesis has been limited due to the difficulty of working with DOT1L protein. For instance, locus-specific or genome-wide binding sites of DOT1L revealed by chromatin immunoprecipitation (ChIP)-based methods are necessary for inferring its functions, but high-quality ChIP-grade antibodies are currently not available. Herein we have developed a knock-in approach to tag endogenous DOT1L with 3 × Flag at its C-terminal domain to follow functional analyses. The knock-in was facilitated by using TALENs to induce a targeted double-strand break at the endogenous DOTIL to stimulate local homologous recombination at that site. The single cell colonies with successful knock-in were isolated and verified by different methods. We also demonstrated that tagged DOT1L maintains its normal function in terms of methylation and that the engineered cells would be very useful for further studies.
