Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-asso...Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-associated virus(AAV)-mediated gene therapy is a promising strategy for treating retinitis pigmentosa.The aim of this study was to explore the molecular mechanisms by which AAV2-PDE6B rescues retinal function.To do this,we injected retinal degeneration 10(rd10)mice subretinally with AAV2-PDE6B and assessed the therapeutic effects on retinal function and structure using dark-and light-adapted electroretinogram,optical coherence tomography,and immunofluorescence.Data-independent acquisition-mass spectrometry-based proteomic analysis was conducted to investigate protein expression levels and pathway enrichment,and the results from this analysis were verified by real-time polymerase chain reaction and western blotting.AAV2-PDE6B injection significantly upregulated PDE6βexpression,preserved electroretinogram responses,and preserved outer nuclear layer thickness in rd10 mice.Differentially expressed proteins between wild-type and rd10 mice were closely related to visual perception,and treating rd10 mice with AAV2-PDE6B restored differentially expressed protein expression to levels similar to those seen in wild-type mice.Kyoto Encyclopedia of Genes and Genome analysis showed that the differentially expressed proteins whose expression was most significantly altered by AAV2-PDE6B injection were enriched in phototransduction pathways.Furthermore,the phototransductionrelated proteins Pde6α,Rom1,Rho,Aldh1a1,and Rbp1 exhibited opposite expression patterns in rd10 mice with or without AAV2-PDE6B treatment.Finally,Bax/Bcl-2,p-ERK/ERK,and p-c-Fos/c-Fos expression levels decreased in rd10 mice following AAV2-PDE6B treatment.Our data suggest that AAV2-PDE6B-mediated gene therapy promotes phototransduction and inhibits apoptosis by inhibiting the ERK signaling pathway and upregulating Bcl-2/Bax expression in retinitis pigmentosa.展开更多
Background:The traditional Sox10 ^(Dom/+)mouse breeding strategy is costly and timeconsuming,so this study aims to optimize the breeding method and improve the scientific research efficiency.Methods:We select the offs...Background:The traditional Sox10 ^(Dom/+)mouse breeding strategy is costly and timeconsuming,so this study aims to optimize the breeding method and improve the scientific research efficiency.Methods:We select the offspring from mating B6C3Fe Sox10 ^(Dom/+)male mice with C57BL/6J female mice,and name the progeny B6C3Fe-g.Further,conduct separate self-breeding for both the B6C3Fe and B6C3Fe-g strains,adhering to the principle of pairing mutants with non-mutants.By comparing the number of offspring,survival rates,and the phenotype of aganglionosis in the colon,a comprehensive evaluation of their breeding capacity and phenotypic stability is conducted.Results:Sanger sequencing results show that the mutation sites of B6C3Fe and B6C3Fe-g mice are consistent.After fluorescent staining of intestinal nerves,it was found that the heterozygous mice of the two strains had neuronal deletion in the distal colon,and this pathological phenotype was consistent with the pathological features of the diseased colon of Hirschsprung disease(HSCR).However,compared with the B6C3Fe strain,the B6C3Fe-g strain has a higher number of offspring and greater survival rates.Conclusions:The breeding strategy of the B6C3Fe-g strain ensures genetic and phenotypic stability,while improving reproductive efficiency,and is an ideal scheme for breeding Sox10 ^(Dom/+)mice.展开更多
目的探讨bcl-2、bcl-6、CD10在弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DL-BCL)中的表达及意义。方法应用免疫组织化学SP法,检测60例弥漫性大B细胞淋巴瘤标本中bcl-2、bcl-6、CD10蛋白的表达水平。结果bcl-2、bcl-6、CD10...目的探讨bcl-2、bcl-6、CD10在弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DL-BCL)中的表达及意义。方法应用免疫组织化学SP法,检测60例弥漫性大B细胞淋巴瘤标本中bcl-2、bcl-6、CD10蛋白的表达水平。结果bcl-2、bcl-6、CD10在60例弥漫性大B细胞淋巴瘤中的表达率分别为55.0%(33/60)、48.3%(29/60)、46.7%(28/60),其中CD10(+)/bcl-6(+)15例,25%;CD10(+)/bcl-6(-)13例,21.7%;CD10(-)/bcl-6(+)14例,23.3%;CD10(-)/bcl-6(-)18例,30%。CD10(+)/bcl-6(+)与CD10(-)/bcl-6(-)中bcl-2的表达差异具有统计学意义。结论bcl-2有望成为DLBCL亚分类的指标。展开更多
目的探讨bcl-2、bcl-6、CD10、MUM1在弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中的表达及与亚分类的关系。方法应用免疫组织化学S-P法,检测60例弥漫性大B细胞淋巴瘤标本中bcl-2、bcl-6、CD10、MUM1蛋白的表达水平。结...目的探讨bcl-2、bcl-6、CD10、MUM1在弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中的表达及与亚分类的关系。方法应用免疫组织化学S-P法,检测60例弥漫性大B细胞淋巴瘤标本中bcl-2、bcl-6、CD10、MUM1蛋白的表达水平。结果bcl-2、bcl-6、CD10、MUM1在60例弥漫性大B细胞淋巴瘤中的表达率分别为55.0%(33/60)、48.3%(29/60)、46.7%(28/60)、58.3%(35/60),其中GCB占55.0%(33/60),non-GCB占45.0%(27/60)。GCB、non-GCB中bcl-2的表达差异具有显著性。在non-GCB中,bcl-6+与bcl-6-的病例bcl-2表达的差异具有显著性。结论该组病例GCB所占的比率超过non-GCB,bcl-2在DLBCL亚分类中可能具有一定的作用。展开更多
基金supported by the National Natural Science Foundation of China,Nos.82071008(to BL)and 82004001(to XJ)Medical Science and Technology Program of Health Commission of Henan Province,No.LHGJ20210072(to RQ)Science and Technology Department of Henan Province,No.212102310307(to XJ)。
文摘Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-associated virus(AAV)-mediated gene therapy is a promising strategy for treating retinitis pigmentosa.The aim of this study was to explore the molecular mechanisms by which AAV2-PDE6B rescues retinal function.To do this,we injected retinal degeneration 10(rd10)mice subretinally with AAV2-PDE6B and assessed the therapeutic effects on retinal function and structure using dark-and light-adapted electroretinogram,optical coherence tomography,and immunofluorescence.Data-independent acquisition-mass spectrometry-based proteomic analysis was conducted to investigate protein expression levels and pathway enrichment,and the results from this analysis were verified by real-time polymerase chain reaction and western blotting.AAV2-PDE6B injection significantly upregulated PDE6βexpression,preserved electroretinogram responses,and preserved outer nuclear layer thickness in rd10 mice.Differentially expressed proteins between wild-type and rd10 mice were closely related to visual perception,and treating rd10 mice with AAV2-PDE6B restored differentially expressed protein expression to levels similar to those seen in wild-type mice.Kyoto Encyclopedia of Genes and Genome analysis showed that the differentially expressed proteins whose expression was most significantly altered by AAV2-PDE6B injection were enriched in phototransduction pathways.Furthermore,the phototransductionrelated proteins Pde6α,Rom1,Rho,Aldh1a1,and Rbp1 exhibited opposite expression patterns in rd10 mice with or without AAV2-PDE6B treatment.Finally,Bax/Bcl-2,p-ERK/ERK,and p-c-Fos/c-Fos expression levels decreased in rd10 mice following AAV2-PDE6B treatment.Our data suggest that AAV2-PDE6B-mediated gene therapy promotes phototransduction and inhibits apoptosis by inhibiting the ERK signaling pathway and upregulating Bcl-2/Bax expression in retinitis pigmentosa.
基金Basic and Applied Basic Research Foundation of Guangxi Province of China,Grant/Award Number:2025GXNSFBA069072National Natural Science Foundation of China,Grant/Award Number:81970450,82070528,82170528,82200561,82201893,82301955,82370526 and 82560108+2 种基金Medical Scientific Research Foundation of Guangdong Province of China,Grant/Award Number:A2024374The Guangdong Basic and Applied Basic Research Foundation,Grant/Award Number:2024A03J1171,2024A03J1238 and 2024A1515013190Science and Technology Projects in Guangzhou,Grant/Award Number:202201020006。
文摘Background:The traditional Sox10 ^(Dom/+)mouse breeding strategy is costly and timeconsuming,so this study aims to optimize the breeding method and improve the scientific research efficiency.Methods:We select the offspring from mating B6C3Fe Sox10 ^(Dom/+)male mice with C57BL/6J female mice,and name the progeny B6C3Fe-g.Further,conduct separate self-breeding for both the B6C3Fe and B6C3Fe-g strains,adhering to the principle of pairing mutants with non-mutants.By comparing the number of offspring,survival rates,and the phenotype of aganglionosis in the colon,a comprehensive evaluation of their breeding capacity and phenotypic stability is conducted.Results:Sanger sequencing results show that the mutation sites of B6C3Fe and B6C3Fe-g mice are consistent.After fluorescent staining of intestinal nerves,it was found that the heterozygous mice of the two strains had neuronal deletion in the distal colon,and this pathological phenotype was consistent with the pathological features of the diseased colon of Hirschsprung disease(HSCR).However,compared with the B6C3Fe strain,the B6C3Fe-g strain has a higher number of offspring and greater survival rates.Conclusions:The breeding strategy of the B6C3Fe-g strain ensures genetic and phenotypic stability,while improving reproductive efficiency,and is an ideal scheme for breeding Sox10 ^(Dom/+)mice.
文摘目的探讨bcl-2、bcl-6、CD10在弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DL-BCL)中的表达及意义。方法应用免疫组织化学SP法,检测60例弥漫性大B细胞淋巴瘤标本中bcl-2、bcl-6、CD10蛋白的表达水平。结果bcl-2、bcl-6、CD10在60例弥漫性大B细胞淋巴瘤中的表达率分别为55.0%(33/60)、48.3%(29/60)、46.7%(28/60),其中CD10(+)/bcl-6(+)15例,25%;CD10(+)/bcl-6(-)13例,21.7%;CD10(-)/bcl-6(+)14例,23.3%;CD10(-)/bcl-6(-)18例,30%。CD10(+)/bcl-6(+)与CD10(-)/bcl-6(-)中bcl-2的表达差异具有统计学意义。结论bcl-2有望成为DLBCL亚分类的指标。
文摘目的探讨bcl-2、bcl-6、CD10、MUM1在弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中的表达及与亚分类的关系。方法应用免疫组织化学S-P法,检测60例弥漫性大B细胞淋巴瘤标本中bcl-2、bcl-6、CD10、MUM1蛋白的表达水平。结果bcl-2、bcl-6、CD10、MUM1在60例弥漫性大B细胞淋巴瘤中的表达率分别为55.0%(33/60)、48.3%(29/60)、46.7%(28/60)、58.3%(35/60),其中GCB占55.0%(33/60),non-GCB占45.0%(27/60)。GCB、non-GCB中bcl-2的表达差异具有显著性。在non-GCB中,bcl-6+与bcl-6-的病例bcl-2表达的差异具有显著性。结论该组病例GCB所占的比率超过non-GCB,bcl-2在DLBCL亚分类中可能具有一定的作用。
