Effective treatment of rheumatoid arthritis can be mediated by native chicken type II collagen(n CCII), recombinant peptide containing n CCII tolerogenic epitopes(CTEs), or a therapeutic DNA vaccine encoding the full-...Effective treatment of rheumatoid arthritis can be mediated by native chicken type II collagen(n CCII), recombinant peptide containing n CCII tolerogenic epitopes(CTEs), or a therapeutic DNA vaccine encoding the full-length CCOL2 A1 c DNA. As recombinant CCII(r CCII) might avoid potential pathogenic virus contamination during n CCII preparation or chromosomal integration and oncogene activation associated with DNA vaccines, here we evaluated the importance of propeptide and telopeptide domains on r CCII triple helix molecular assembly. We constructed p C-and p N-procollagen(without N-or Cpropeptides, respectively) as well as CTEs located in the triple helical domain lacking both propeptides and telopeptides, and expressed these in yeast Pichia pastoris host strain GS115(his4, Mut+) simultaneously with recombinant chicken prolyl-4-hydroxylase α and β subunits. Both p C-and p N-procollagen monomers accumulated inside P. pastoris cells, whereas CTE was assembled into homotrimers with stable conformation and secreted into the supernatants, suggesting that the large molecular weight p C-or p N-procollagens were retained within the endoplasmic reticulum whereas the smaller CTEs proceeded through the secretory pathway. Furthermore, resulting recombinant chicken type II collagen p Cα1(II) can induce collagen-induced arthritis(CIA) rat model, which seems to be as effective as the current standard n CCII. Notably, protease digestion assays showed that r CCII could assemble in the absence of C-and N-propeptides or telopeptides. These findings provide new insights into the minimal structural requirements for r CCII expression and folding.展开更多
The world is currently facing a highly contagious and fatal pandemic caused by SARS-CoV-2,which is also the first pandemic caused by a coronavirus yet.As of 25 August 2020,the number of confirmed COVID-19 patients has...The world is currently facing a highly contagious and fatal pandemic caused by SARS-CoV-2,which is also the first pandemic caused by a coronavirus yet.As of 25 August 2020,the number of confirmed COVID-19 patients has exceeded 23,518,343 worldwide,with a death toll of over 810,492.1 To date,the greatest challenge we face is the incomplete knowledge of both SARS-CoV-2 and COVID-19,resulting in a lack of effective drugs and adequate treatment strategies.Despite various treatment options for critical COVID-19 patients,including extracorporeal membrane oxygenation,ventilation,tracheal intubation,circulatory support,and non-specific antiviral drugs,clinical outcomes are unsatisfactory.2 In particular,some mild or common COVID-19 patients experience sudden and rapid deterioration with the onset of fatal cytokine storm syndrome which cannot be treated by any conventional therapies.Therefore,from 2 February 2020 to 4 March 2020,the China National Health Commission has issued successively the 1st–7th editions of the Official Guidance of Diagnosis and Management for COVID-19 and formulated preliminary guidelines for the clinical application of convalescent plasma therapy(CPT)for critical COVID-19 patients.展开更多
The major histocompatibility complex(MHC)is closely associated with numerous diseases,but its high degree of polymorphism complicates the discovery of disease-associated variants.In principle,recombination and de novo...The major histocompatibility complex(MHC)is closely associated with numerous diseases,but its high degree of polymorphism complicates the discovery of disease-associated variants.In principle,recombination and de novo mutations are two critical factors responsible for MHC polymorphisms.However,direct evidence for this hypothesis is lacking.Here,we report the generation of fine-scale MHC recombination and de novo mutation maps of~5 Mb by deep sequencing(>100×)of the MHC genome for 17 MHC recombination and 30 non-recombination Han Chinese families(a total of 190 individuals).Recombination hotspots and Han-specific breakpoints are located in close proximity at haplotype block boundaries.The average MHC de novo mutation rate is higher than the genome-wide de novo mutation rate,particularly in MHC recombinant individuals.Notably,mutation and recombination generated polymorphisms are located within and outside linkage disequilibrium regions of the MHC,respectively,and evolution of the MHC locus was mainly controlled by positive selection.These findings provide insights on the evolutionary causes of the MHC diversity and may facilitate the identification of disease-associated genetic variants.展开更多
文摘Effective treatment of rheumatoid arthritis can be mediated by native chicken type II collagen(n CCII), recombinant peptide containing n CCII tolerogenic epitopes(CTEs), or a therapeutic DNA vaccine encoding the full-length CCOL2 A1 c DNA. As recombinant CCII(r CCII) might avoid potential pathogenic virus contamination during n CCII preparation or chromosomal integration and oncogene activation associated with DNA vaccines, here we evaluated the importance of propeptide and telopeptide domains on r CCII triple helix molecular assembly. We constructed p C-and p N-procollagen(without N-or Cpropeptides, respectively) as well as CTEs located in the triple helical domain lacking both propeptides and telopeptides, and expressed these in yeast Pichia pastoris host strain GS115(his4, Mut+) simultaneously with recombinant chicken prolyl-4-hydroxylase α and β subunits. Both p C-and p N-procollagen monomers accumulated inside P. pastoris cells, whereas CTE was assembled into homotrimers with stable conformation and secreted into the supernatants, suggesting that the large molecular weight p C-or p N-procollagens were retained within the endoplasmic reticulum whereas the smaller CTEs proceeded through the secretory pathway. Furthermore, resulting recombinant chicken type II collagen p Cα1(II) can induce collagen-induced arthritis(CIA) rat model, which seems to be as effective as the current standard n CCII. Notably, protease digestion assays showed that r CCII could assemble in the absence of C-and N-propeptides or telopeptides. These findings provide new insights into the minimal structural requirements for r CCII expression and folding.
文摘The world is currently facing a highly contagious and fatal pandemic caused by SARS-CoV-2,which is also the first pandemic caused by a coronavirus yet.As of 25 August 2020,the number of confirmed COVID-19 patients has exceeded 23,518,343 worldwide,with a death toll of over 810,492.1 To date,the greatest challenge we face is the incomplete knowledge of both SARS-CoV-2 and COVID-19,resulting in a lack of effective drugs and adequate treatment strategies.Despite various treatment options for critical COVID-19 patients,including extracorporeal membrane oxygenation,ventilation,tracheal intubation,circulatory support,and non-specific antiviral drugs,clinical outcomes are unsatisfactory.2 In particular,some mild or common COVID-19 patients experience sudden and rapid deterioration with the onset of fatal cytokine storm syndrome which cannot be treated by any conventional therapies.Therefore,from 2 February 2020 to 4 March 2020,the China National Health Commission has issued successively the 1st–7th editions of the Official Guidance of Diagnosis and Management for COVID-19 and formulated preliminary guidelines for the clinical application of convalescent plasma therapy(CPT)for critical COVID-19 patients.
基金supported by grants from the National Key Basic Research Development Program of China(grants No.2009CB522401 and 2003CB515509,and AWS14C014)。
文摘The major histocompatibility complex(MHC)is closely associated with numerous diseases,but its high degree of polymorphism complicates the discovery of disease-associated variants.In principle,recombination and de novo mutations are two critical factors responsible for MHC polymorphisms.However,direct evidence for this hypothesis is lacking.Here,we report the generation of fine-scale MHC recombination and de novo mutation maps of~5 Mb by deep sequencing(>100×)of the MHC genome for 17 MHC recombination and 30 non-recombination Han Chinese families(a total of 190 individuals).Recombination hotspots and Han-specific breakpoints are located in close proximity at haplotype block boundaries.The average MHC de novo mutation rate is higher than the genome-wide de novo mutation rate,particularly in MHC recombinant individuals.Notably,mutation and recombination generated polymorphisms are located within and outside linkage disequilibrium regions of the MHC,respectively,and evolution of the MHC locus was mainly controlled by positive selection.These findings provide insights on the evolutionary causes of the MHC diversity and may facilitate the identification of disease-associated genetic variants.