AIM: To study the role of autophagy and the relationship between retinoic acid receptor α(RARα) and autophagy in liver ischemia and reperfusion(IR) injury.METHODS: All-trans retinoic acid(ATRA) was administered to m...AIM: To study the role of autophagy and the relationship between retinoic acid receptor α(RARα) and autophagy in liver ischemia and reperfusion(IR) injury.METHODS: All-trans retinoic acid(ATRA) was administered to mice for two weeks before operation. Reverse transcription-polymerase chain reaction and Western blot were used to detect the expression levels of related factors. To demonstrate the role of RARα,LE540,a RARα inhibitor,was used to treat hepatocytes injured by H2O2 in vitro.RESULTS: ATRA pretreatment noticeably diminished levels of serum alanine aminotransferase and as-partate aminotransferase as well as the degree of histopathological changes. Apoptosis was also inhibited,whereas autophagy was promoted. In vitro,RARα was inhibited by LE540,which resulted in decreased autophagy and increased apoptosis. Similarly,the expression of Foxo3 a and p-Akt was downregulated,but Foxo1 expression was upregulated.CONCLUSION: This research provides evidence that ATRA can protect the liver from IR injury by promoting autophagy,which is dependent on Foxo3/p-Akt/Foxo1 signaling.展开更多
AIM: To perform sequencing analysis in patients with very early-onset inflammatory bowel disease (VEO-IBD) to determine the genetic basis for VEO-IBD in Chinese pediatric patients. METHODS: A total of 13 Chinese pedia...AIM: To perform sequencing analysis in patients with very early-onset inflammatory bowel disease (VEO-IBD) to determine the genetic basis for VEO-IBD in Chinese pediatric patients. METHODS: A total of 13 Chinese pediatric patients with VEO-IBD were diagnosed from May 2012 and August 2014. The relevant clinical characteristics of these patients were analyzed. Then DNA in the peripheral blood from patients was extracted. Next generation sequencing (NGS) based on an Illumina-Miseq platform was used to analyze the exons in the coding regions of 10 candidate genes: IL-10, IL-10RA, IL-10RB, NOD2, FUT2, IL23R, GPR35, GPR65, TNFSF15, and ADAM30. The Sanger sequencing was used to verify the variations detected in NGS. RESULTS: Out of the 13 pediatric patients, ten were diagnosed with Crohn's disease, and three diagnosed with ulcerative colitis. Mutations in IL-10RA and IL-10RB were detected in five patients. There were four patients who had single nucleotide polymorphisms associated with IBD. Two patients had IL-10RA and FUT2 polymorphisms, and two patients had IL-10RB and FUT2 polymorphisms. Gene variations were not found in the rest four patients. Children with mutations had lower percentile body weight ( 1.0% vs 27.5%, P = 0.002) and hemoglobin ( 87.4 g/L vs 108.5 g/L, P = 0.040) when compared with children without mutations. Although the age of onset was earlier, height was shorter, and the response to treatment was poorer in the mutation group, there was no significant difference in these factors between groups. CONCLUSION: IL-10RA and IL-10RB mutations are common in Chinese children with VEO-IBD. Patients with mutations have an earlier disease onset, lower body weight and hemoglobin, and poorer展开更多
Recombineering is an essential tool for molecular biologists,allowing for the facile and efficient manipulation of bacterial genomes directly in cells without the need for costly and laborious in vitro manipulations i...Recombineering is an essential tool for molecular biologists,allowing for the facile and efficient manipulation of bacterial genomes directly in cells without the need for costly and laborious in vitro manipulations involving restriction enzymes.The main workhorses behind recombineering are bacteriophage proteins that promote the single-strand annealing(SSA)homologous recombination pathway to repair double-stranded DNA breaks.While there have been several reviews examining recombineering methods and applications,comparatively few have focused on the mechanisms of the proteins that are the key players in the SSA pathway:a 5′→3′exonuclease and a single-strand annealing protein(SSAP or“annealase”).This review dives into the structures and functions of the two SSA recombination systems that were the first to be developed for recombineering in E.coli:the RecET system from E.coli Rac prophage and the𝜆Red system from bacteriophageλ.By comparing the structures of the RecT and Red𝛽annealases,and the RecE and𝜆Exo exonucleases,we provide new insights into how the structures of these proteins dictate their function.Examining the sequence conservation of the𝜆λExo and RecE exonucleases gives more profound insights into their critical functional features.Ultimately,as recombineering accelerates and evolves in the laboratory,a better understanding of the mechanisms of the proteins behind this powerful technique will drive the development of improved and expanded capabilities in the future.展开更多
基金Supported by National Natural Science Foundation of ChinaNo.81273261
文摘AIM: To study the role of autophagy and the relationship between retinoic acid receptor α(RARα) and autophagy in liver ischemia and reperfusion(IR) injury.METHODS: All-trans retinoic acid(ATRA) was administered to mice for two weeks before operation. Reverse transcription-polymerase chain reaction and Western blot were used to detect the expression levels of related factors. To demonstrate the role of RARα,LE540,a RARα inhibitor,was used to treat hepatocytes injured by H2O2 in vitro.RESULTS: ATRA pretreatment noticeably diminished levels of serum alanine aminotransferase and as-partate aminotransferase as well as the degree of histopathological changes. Apoptosis was also inhibited,whereas autophagy was promoted. In vitro,RARα was inhibited by LE540,which resulted in decreased autophagy and increased apoptosis. Similarly,the expression of Foxo3 a and p-Akt was downregulated,but Foxo1 expression was upregulated.CONCLUSION: This research provides evidence that ATRA can protect the liver from IR injury by promoting autophagy,which is dependent on Foxo3/p-Akt/Foxo1 signaling.
基金Supported by National Nature Science Foundation of China,No.81400588
文摘AIM: To perform sequencing analysis in patients with very early-onset inflammatory bowel disease (VEO-IBD) to determine the genetic basis for VEO-IBD in Chinese pediatric patients. METHODS: A total of 13 Chinese pediatric patients with VEO-IBD were diagnosed from May 2012 and August 2014. The relevant clinical characteristics of these patients were analyzed. Then DNA in the peripheral blood from patients was extracted. Next generation sequencing (NGS) based on an Illumina-Miseq platform was used to analyze the exons in the coding regions of 10 candidate genes: IL-10, IL-10RA, IL-10RB, NOD2, FUT2, IL23R, GPR35, GPR65, TNFSF15, and ADAM30. The Sanger sequencing was used to verify the variations detected in NGS. RESULTS: Out of the 13 pediatric patients, ten were diagnosed with Crohn's disease, and three diagnosed with ulcerative colitis. Mutations in IL-10RA and IL-10RB were detected in five patients. There were four patients who had single nucleotide polymorphisms associated with IBD. Two patients had IL-10RA and FUT2 polymorphisms, and two patients had IL-10RB and FUT2 polymorphisms. Gene variations were not found in the rest four patients. Children with mutations had lower percentile body weight ( 1.0% vs 27.5%, P = 0.002) and hemoglobin ( 87.4 g/L vs 108.5 g/L, P = 0.040) when compared with children without mutations. Although the age of onset was earlier, height was shorter, and the response to treatment was poorer in the mutation group, there was no significant difference in these factors between groups. CONCLUSION: IL-10RA and IL-10RB mutations are common in Chinese children with VEO-IBD. Patients with mutations have an earlier disease onset, lower body weight and hemoglobin, and poorer
基金the National Science Foundation Grant MCB-2212951(to CEB)and NHMRC Ideas grant APP1184012/GNT1184012(to GT).
文摘Recombineering is an essential tool for molecular biologists,allowing for the facile and efficient manipulation of bacterial genomes directly in cells without the need for costly and laborious in vitro manipulations involving restriction enzymes.The main workhorses behind recombineering are bacteriophage proteins that promote the single-strand annealing(SSA)homologous recombination pathway to repair double-stranded DNA breaks.While there have been several reviews examining recombineering methods and applications,comparatively few have focused on the mechanisms of the proteins that are the key players in the SSA pathway:a 5′→3′exonuclease and a single-strand annealing protein(SSAP or“annealase”).This review dives into the structures and functions of the two SSA recombination systems that were the first to be developed for recombineering in E.coli:the RecET system from E.coli Rac prophage and the𝜆Red system from bacteriophageλ.By comparing the structures of the RecT and Red𝛽annealases,and the RecE and𝜆Exo exonucleases,we provide new insights into how the structures of these proteins dictate their function.Examining the sequence conservation of the𝜆λExo and RecE exonucleases gives more profound insights into their critical functional features.Ultimately,as recombineering accelerates and evolves in the laboratory,a better understanding of the mechanisms of the proteins behind this powerful technique will drive the development of improved and expanded capabilities in the future.
