AIM To stably correct tyrosinaemia in proliferating livers of fumarylacetoacetate-hydrolase knockout(Fah-/-) mice by homologous-recombination-mediated targeted addition of the Fah gene.METHODS C57 BL/6 Fah?exon5 mice ...AIM To stably correct tyrosinaemia in proliferating livers of fumarylacetoacetate-hydrolase knockout(Fah-/-) mice by homologous-recombination-mediated targeted addition of the Fah gene.METHODS C57 BL/6 Fah?exon5 mice served as an animal model for human tyrosinaemia type 1 in our study. The vector was created by amplifying human Fah c DNA including the TTR promoter from a lentivirus plasmid as described. The Fah expression cassette was flanked by homologous arms(620 bp and 749 bp long) of the Rosa26 gene locus. Mice were injected with 2.1 × 108 VP of this vector(r AAV8-ROSA26.HAL-TTR.FahROSA26.HAR) via the tail vein. Mice in the control group were injected with 2.1 × 108 VP of a similar vector but missing the homologous arms(r AAV8-TTR.Fah). Primary hepatocytes from Fah-/-recipient mice, treated with our vectors, were isolated and 1 × 106 hepatocytes were transplanted into secondary Fah-/-recipient mice by injection into the spleen. Upon either vector application or hepatocyte transplantation NTBC treatment was stopped in recipient mice. RESULTS Here, we report successful HR-mediated genome editing by integration of a Fah gene expression cassette into the "safe harbour locus" Rosa26 by recombinant AAV8. Both groups of mice showed long-term survival, weight gain and FAH positive clusters as determined by immunohistochemistry analysis of liver sections in the absence of NTBC treatment. In the group of C57 BL/6 Fah?exon5 mice, which have been transplanted with hepatocytes from a mouse injected with r AAV8-ROSA26.HAL-TTR.Fah-ROSA26.HAR 156 d before, 6 out of 6 mice showed long-term survival, weight gain and FAH positive clusters without need for NTBC treatment. In contrast only 1 out 5 mice, who received hepatocytes from r AAV8-TTR.Fah treated mice, survived and showed few and smaller FAH positive clusters. These results demonstrate that homologous recombinationmediated Fah gene transfer corrects the phenotype in a mouse model of human tyrosinaemia type 1(Fah-/-mice) and is long lasting in a proliferating state of 展开更多
This paper draws attention to the issue of the vibration absorption of nonlinear mechani- cal system coupled to nonlinear energy sink （NES） under the impact of the narrow band stochastic excitation. Firstly, based o...This paper draws attention to the issue of the vibration absorption of nonlinear mechani- cal system coupled to nonlinear energy sink （NES） under the impact of the narrow band stochastic excitation. Firstly, based on the complex-averaging method and frequency detuning methodology, response regimes of oscillators have been researched under the linear impact of coupling a nonlinear attachment with less relativistic mass and an external sinusoidal forcing, of which results turn out that the quasi-periodicity response regime of system which occurs when the external excitation amplitude exceeds the critical values will be the precondition of the targeted energy transfer. Secondly, basing on the path integration method, vibration suppression of NES has been researched when it is affected by a main oscillator with a narrow band stochastic force in the form of trigono- metric functions, of which results show that response regimes are affected by the amplitude of stochastic excitation and the disturbance strength. Finally, all these conclusions have been approved by the numerical verification and coincided with the theoretical analysis; meanwhile, after the com- paring analysis with the optimal linear absorber, it turns out that the NES which is affected by the narrow band stochastic force could also suppress the vibration of system with a better effect.展开更多
基金Rebirth, SFB 738the "Deutsche Forschungsgemeinschaft" (Gerok-Grant) for financial support
文摘AIM To stably correct tyrosinaemia in proliferating livers of fumarylacetoacetate-hydrolase knockout(Fah-/-) mice by homologous-recombination-mediated targeted addition of the Fah gene.METHODS C57 BL/6 Fah?exon5 mice served as an animal model for human tyrosinaemia type 1 in our study. The vector was created by amplifying human Fah c DNA including the TTR promoter from a lentivirus plasmid as described. The Fah expression cassette was flanked by homologous arms(620 bp and 749 bp long) of the Rosa26 gene locus. Mice were injected with 2.1 × 108 VP of this vector(r AAV8-ROSA26.HAL-TTR.FahROSA26.HAR) via the tail vein. Mice in the control group were injected with 2.1 × 108 VP of a similar vector but missing the homologous arms(r AAV8-TTR.Fah). Primary hepatocytes from Fah-/-recipient mice, treated with our vectors, were isolated and 1 × 106 hepatocytes were transplanted into secondary Fah-/-recipient mice by injection into the spleen. Upon either vector application or hepatocyte transplantation NTBC treatment was stopped in recipient mice. RESULTS Here, we report successful HR-mediated genome editing by integration of a Fah gene expression cassette into the "safe harbour locus" Rosa26 by recombinant AAV8. Both groups of mice showed long-term survival, weight gain and FAH positive clusters as determined by immunohistochemistry analysis of liver sections in the absence of NTBC treatment. In the group of C57 BL/6 Fah?exon5 mice, which have been transplanted with hepatocytes from a mouse injected with r AAV8-ROSA26.HAL-TTR.Fah-ROSA26.HAR 156 d before, 6 out of 6 mice showed long-term survival, weight gain and FAH positive clusters without need for NTBC treatment. In contrast only 1 out 5 mice, who received hepatocytes from r AAV8-TTR.Fah treated mice, survived and showed few and smaller FAH positive clusters. These results demonstrate that homologous recombinationmediated Fah gene transfer corrects the phenotype in a mouse model of human tyrosinaemia type 1(Fah-/-mice) and is long lasting in a proliferating state of
基金supported by the National Natural Science Foundation of China (No. 51375109)
文摘This paper draws attention to the issue of the vibration absorption of nonlinear mechani- cal system coupled to nonlinear energy sink （NES） under the impact of the narrow band stochastic excitation. Firstly, based on the complex-averaging method and frequency detuning methodology, response regimes of oscillators have been researched under the linear impact of coupling a nonlinear attachment with less relativistic mass and an external sinusoidal forcing, of which results turn out that the quasi-periodicity response regime of system which occurs when the external excitation amplitude exceeds the critical values will be the precondition of the targeted energy transfer. Secondly, basing on the path integration method, vibration suppression of NES has been researched when it is affected by a main oscillator with a narrow band stochastic force in the form of trigono- metric functions, of which results show that response regimes are affected by the amplitude of stochastic excitation and the disturbance strength. Finally, all these conclusions have been approved by the numerical verification and coincided with the theoretical analysis; meanwhile, after the com- paring analysis with the optimal linear absorber, it turns out that the NES which is affected by the narrow band stochastic force could also suppress the vibration of system with a better effect.
