Mutations of PTEN-induced kinase I(PINK1)cause early-onset Parkinson’s disease(PD)with selective neurodegeneration in humans.However,current PINK1 knockout mouse and pig models are unable to recapitulate the typical ...Mutations of PTEN-induced kinase I(PINK1)cause early-onset Parkinson’s disease(PD)with selective neurodegeneration in humans.However,current PINK1 knockout mouse and pig models are unable to recapitulate the typical neurodegenerative phenotypes observed in PD patients.This suggests that generating PINK1 disease models in non-human primates(NHPs)that are close to humans is essential to investigate the unique function of PINK1 in primate brains.Paired single guide RNA(sgRNA)/Cas9-D10A nickases and truncated sgRNA/Cas9,both of which can reduce off-target effects without compromising on-target editing,are two optimized strategies in the CRISPR/Cas9 system for establishing disease animal models.Here,we combined the two strategies and injected Cas9-D10A mRNA and two truncated sgRNAs into one-cell-stage cynomolgus zygotes to target the PINK1 gene.We achieved precise and efficient gene editing of the target site in three newborn cynomolgus monkeys.The frame shift mutations of PINK1 in mutant fibroblasts led to a reduction in mRNA.However,western blotting and immunofluorescence staining confirmed the PINK1 protein levels were comparable to that in wild-type fibroblasts.We further reprogramed mutant fibroblasts into induced pluripotent stem cells(iPSCs),which showed similar ability to differentiate into dopamine(DA)neurons.Taken together,our results showed that co-injection of Cas9-D10A nickase mRNA and sgRNA into one-cell-stage cynomolgus embryos enabled the generation of human disease models in NHPs and target editing by pair truncated sgRNA/Cas9-D10A in PINK1 gene exon 2 did not impact protein expression.展开更多
Experiments were conducted to study the temperature field,flow field,3-D characteristics and self-sustained oscillation of the natural air convection in a 3-D shallow cavity which was heated from underneath. The exper...Experiments were conducted to study the temperature field,flow field,3-D characteristics and self-sustained oscillation of the natural air convection in a 3-D shallow cavity which was heated from underneath. The experiments were conducted by the methods of laser holographic interferometry photography and smoke visualization. To ensure the temperature of both plates to be constant and the accuracy of laser interferometer,the instrument was calibrated and error has was analyzed. The results showed that the flow field was stable at lower Rayleigh numbers. When the Rayleigh number increased,the flow field became instable and the isotherms distorted. The rolls merged at Ra=12500 and formed along both axes when Ra was over 18500. The air rose from the middle and descended in the circumference while the flow field and heat transfer converted to 3-D characteristic from 2-D characteristic. When the Rayleigh number increased,the flow field became more instable. The rolls became irregular and time dependent when Ra≥Rac(=30500) ,which is nonlinear. When lateral walls were heated or cooled,the rolls merged along the long axis and two rolls formed along the short axis. Three rolls formed occasionally in the process.展开更多
In this paper,we discuss the mathematical relation determined by the basic physical constant between three types of quarks and the masses of leptons with charges in detail.First,by further theoretical analysis,we wond...In this paper,we discuss the mathematical relation determined by the basic physical constant between three types of quarks and the masses of leptons with charges in detail.First,by further theoretical analysis,we wonderfully see that the result got from the mass empirical formula of quark and charged lepton is identical with the data received by theoretical estimating from the gauge field theory.Second,we also gladly see that the result got from quark and lepton(with charges)mass empirical formula is completely accordant with experiment data.These mass formulas reveal the essential relation between me,mμ,mτ and mq.At the same time,the empirical formula may also derive the mass formula of neu-trinos.As to the mass of neutrinos,at present we only know the square difference of its mass,and so this is meaningful to theoretically estimating the mass.展开更多
基金supported by the National Key Research and Development Program(2016YFA0101401 and 2018YFA0801400)Major Basic Research Project of Science and Technology of Yunnan(2019FY002 and 202001BC070001)。
文摘Mutations of PTEN-induced kinase I(PINK1)cause early-onset Parkinson’s disease(PD)with selective neurodegeneration in humans.However,current PINK1 knockout mouse and pig models are unable to recapitulate the typical neurodegenerative phenotypes observed in PD patients.This suggests that generating PINK1 disease models in non-human primates(NHPs)that are close to humans is essential to investigate the unique function of PINK1 in primate brains.Paired single guide RNA(sgRNA)/Cas9-D10A nickases and truncated sgRNA/Cas9,both of which can reduce off-target effects without compromising on-target editing,are two optimized strategies in the CRISPR/Cas9 system for establishing disease animal models.Here,we combined the two strategies and injected Cas9-D10A mRNA and two truncated sgRNAs into one-cell-stage cynomolgus zygotes to target the PINK1 gene.We achieved precise and efficient gene editing of the target site in three newborn cynomolgus monkeys.The frame shift mutations of PINK1 in mutant fibroblasts led to a reduction in mRNA.However,western blotting and immunofluorescence staining confirmed the PINK1 protein levels were comparable to that in wild-type fibroblasts.We further reprogramed mutant fibroblasts into induced pluripotent stem cells(iPSCs),which showed similar ability to differentiate into dopamine(DA)neurons.Taken together,our results showed that co-injection of Cas9-D10A nickase mRNA and sgRNA into one-cell-stage cynomolgus embryos enabled the generation of human disease models in NHPs and target editing by pair truncated sgRNA/Cas9-D10A in PINK1 gene exon 2 did not impact protein expression.
基金supported by the National Natural Science Foundation of China (Grant No. 50978120)the Shanghai Leading Academic Discipline Project (Grant No. 05EZ18)the Doctoral Fund Project of Jilin Institute of Architectural and Civil Engineering (Grant No. 861055)
文摘Experiments were conducted to study the temperature field,flow field,3-D characteristics and self-sustained oscillation of the natural air convection in a 3-D shallow cavity which was heated from underneath. The experiments were conducted by the methods of laser holographic interferometry photography and smoke visualization. To ensure the temperature of both plates to be constant and the accuracy of laser interferometer,the instrument was calibrated and error has was analyzed. The results showed that the flow field was stable at lower Rayleigh numbers. When the Rayleigh number increased,the flow field became instable and the isotherms distorted. The rolls merged at Ra=12500 and formed along both axes when Ra was over 18500. The air rose from the middle and descended in the circumference while the flow field and heat transfer converted to 3-D characteristic from 2-D characteristic. When the Rayleigh number increased,the flow field became more instable. The rolls became irregular and time dependent when Ra≥Rac(=30500) ,which is nonlinear. When lateral walls were heated or cooled,the rolls merged along the long axis and two rolls formed along the short axis. Three rolls formed occasionally in the process.
基金the State Key Basic Research Project(973Project)of China(Grant No.2006CB605102)the National Natural Science Foundation of China(Grant No.90306016)
文摘In this paper,we discuss the mathematical relation determined by the basic physical constant between three types of quarks and the masses of leptons with charges in detail.First,by further theoretical analysis,we wonderfully see that the result got from the mass empirical formula of quark and charged lepton is identical with the data received by theoretical estimating from the gauge field theory.Second,we also gladly see that the result got from quark and lepton(with charges)mass empirical formula is completely accordant with experiment data.These mass formulas reveal the essential relation between me,mμ,mτ and mq.At the same time,the empirical formula may also derive the mass formula of neu-trinos.As to the mass of neutrinos,at present we only know the square difference of its mass,and so this is meaningful to theoretically estimating the mass.
