Neurotrophin-3 (NT-3) can promote the repair of central nervous system and retinal damage. In previous reports, NT-3 has been expressed by viral vectors. However, plasmid vectors have a safer profile compared with vir...Neurotrophin-3 (NT-3) can promote the repair of central nervous system and retinal damage. In previous reports, NT-3 has been expressed by viral vectors. However, plasmid vectors have a safer profile compared with viral vectors in clinical studies. This study recombined amplified human retinal NT-3 with a eukaryotic expression plasmid containing green fluorescent protein (GFP) to construct an NT-3 expression plasmid, pEGFP-N1-NT-3. The transfection efficiency 48 hours after pEGFP-N1-NT-3 transfection to 293T cells was 50.06 ± 2.78%. Abundant NT-3-GFP was expressed in 293T cells as observed by fluorescence microscopy, suggesting the construct pEGFP-N1-NT-3 effectively expressed and secreted NT-3-GFP. Secretory vesicles containing NT-3-GFP were observed in a constant location in cells by laser scan confocal microscopy, indicating the expression and secretion processes of NT-3 in eukaryotic cells were in accordance with the physical synthesis processes of secreted proteins. Western blot assay showed that pro-NT-3-GFP had a molecular weight of 56 kDa, further confirming NT-3-GFP expression. At 48 hours after transfection, the concentration of NT-3 in culture medium was 22.3 ng/mL, suggesting NT-3 produced by pEGFP-N1-NT-3 was efficiently secreted. This study constructed a human retinal-derived NT-3 eukaryotic expression plasmid that efficiently expressed and secreted NT-3.展开更多
The rare disease of chronic infantile neurological cutaneous and articular(CINCA)syndrome,is caused by the over-secretion of interleukin(IL)-1βdue to a gain-of-function NLRP3 gene mutation in the autosomal chromosome...The rare disease of chronic infantile neurological cutaneous and articular(CINCA)syndrome,is caused by the over-secretion of interleukin(IL)-1βdue to a gain-of-function NLRP3 gene mutation in the autosomal chromosome which often involves in eyes.In this report,we studied a 9-year-old girl with CINCA.The eyes were also involved and presented bilateral papilledema.Genetic testing revealed that the symptoms were caused by a novel gene mutation site(c.913G>A,p.D305N)in conservative domain exon-3 of NLRP3 which is gain-function gene of CINCA.The patient had the characteristic facial features,frontal fossa and saddle nose,manifested the generalized urticaria-like skin rash at two weeks after birth,periodic fever 6 months after birth,sensorineural deafness at 7 years old,and bilateral papilledema,aseptic meningitis and knee arthropathy at 9 years old.White cell counts,C-reactive protein increased and intracranial pressure raised to 300 mmH2O.The meningeal thickening enhanced by gadolinium in magnetic resonance imaging(MRI).Based on clinical features and genetic test,the girl was diagnosed bilateral papilledema secondary to CINCA and administered prednisone and lowered intracranial pressure medicine to resolve symptoms.With 3-year follow-up,patient had no inflammatory flare-up with visual acuity improvement.The finding of novel genetic mutation site(p.D305N)in NLRP3 gene expanded genotype spectrum associated with CINCA.This case also expanded the cause spectrum of papilledema and it highlighted systemic disease history for patients with bilateral papilledema.展开更多
Background:To evaluate the feature of different retinal layer segmentation in neuromyelitis optica spectrum disorders(NMOSD)with spectral-domain optical coherence tomography(SD-OCT)and to compare it with that in multi...Background:To evaluate the feature of different retinal layer segmentation in neuromyelitis optica spectrum disorders(NMOSD)with spectral-domain optical coherence tomography(SD-OCT)and to compare it with that in multiple sclerosis(MS),healthy controls(HC),and idiopathic optic neuritis(ION).Methods:We retrieved four electronic databases,including Pubmed,Embase,Cochrane Library,and Web of Science from inception to September 1st,2021.A meta-analysis was performed to compare different retinal layer segmentation thicknesses between patients with or without a history of optic neuritis(ON)in NMOSD and the control group,including patients with MS,HC,and ION.Results:Forty-two studies were included and the interval between the last ON onset and examination was greater than 3 months.Compared with that in HC eyes,the loss of retinal nerve fiber layer(RNFL)and macular ganglion cell and inner plexiform layer(GC-IPL)was serious in NMOSD eye especially after ON.Moreover,compared with that in ION eyes or MS-related-ON eyes,the injury to the peripapillary retinal nerve fiber layer(pRNFL)was severe in NMOSD-related-ON eyes.In addition,the correlation coefficient between pRNFL and prognostic visual acuity was 0.43.However,the one-arm study revealed the inner nuclear layer(INL)was thickened in NMOSDrelated-ON eyes compared with HC eyes.Conclusions:Inclusion of the RNFL and macular GC-IPL is recommended for monitoring disease progression and attention should be paid to changes in the INL.展开更多
基金supported by the National Natural Science Foundation of China, No. 30973262
文摘Neurotrophin-3 (NT-3) can promote the repair of central nervous system and retinal damage. In previous reports, NT-3 has been expressed by viral vectors. However, plasmid vectors have a safer profile compared with viral vectors in clinical studies. This study recombined amplified human retinal NT-3 with a eukaryotic expression plasmid containing green fluorescent protein (GFP) to construct an NT-3 expression plasmid, pEGFP-N1-NT-3. The transfection efficiency 48 hours after pEGFP-N1-NT-3 transfection to 293T cells was 50.06 ± 2.78%. Abundant NT-3-GFP was expressed in 293T cells as observed by fluorescence microscopy, suggesting the construct pEGFP-N1-NT-3 effectively expressed and secreted NT-3-GFP. Secretory vesicles containing NT-3-GFP were observed in a constant location in cells by laser scan confocal microscopy, indicating the expression and secretion processes of NT-3 in eukaryotic cells were in accordance with the physical synthesis processes of secreted proteins. Western blot assay showed that pro-NT-3-GFP had a molecular weight of 56 kDa, further confirming NT-3-GFP expression. At 48 hours after transfection, the concentration of NT-3 in culture medium was 22.3 ng/mL, suggesting NT-3 produced by pEGFP-N1-NT-3 was efficiently secreted. This study constructed a human retinal-derived NT-3 eukaryotic expression plasmid that efficiently expressed and secreted NT-3.
文摘The rare disease of chronic infantile neurological cutaneous and articular(CINCA)syndrome,is caused by the over-secretion of interleukin(IL)-1βdue to a gain-of-function NLRP3 gene mutation in the autosomal chromosome which often involves in eyes.In this report,we studied a 9-year-old girl with CINCA.The eyes were also involved and presented bilateral papilledema.Genetic testing revealed that the symptoms were caused by a novel gene mutation site(c.913G>A,p.D305N)in conservative domain exon-3 of NLRP3 which is gain-function gene of CINCA.The patient had the characteristic facial features,frontal fossa and saddle nose,manifested the generalized urticaria-like skin rash at two weeks after birth,periodic fever 6 months after birth,sensorineural deafness at 7 years old,and bilateral papilledema,aseptic meningitis and knee arthropathy at 9 years old.White cell counts,C-reactive protein increased and intracranial pressure raised to 300 mmH2O.The meningeal thickening enhanced by gadolinium in magnetic resonance imaging(MRI).Based on clinical features and genetic test,the girl was diagnosed bilateral papilledema secondary to CINCA and administered prednisone and lowered intracranial pressure medicine to resolve symptoms.With 3-year follow-up,patient had no inflammatory flare-up with visual acuity improvement.The finding of novel genetic mutation site(p.D305N)in NLRP3 gene expanded genotype spectrum associated with CINCA.This case also expanded the cause spectrum of papilledema and it highlighted systemic disease history for patients with bilateral papilledema.
基金supported by China-USA intergovernmental Cooperation program(2018YFE0113900).
文摘Background:To evaluate the feature of different retinal layer segmentation in neuromyelitis optica spectrum disorders(NMOSD)with spectral-domain optical coherence tomography(SD-OCT)and to compare it with that in multiple sclerosis(MS),healthy controls(HC),and idiopathic optic neuritis(ION).Methods:We retrieved four electronic databases,including Pubmed,Embase,Cochrane Library,and Web of Science from inception to September 1st,2021.A meta-analysis was performed to compare different retinal layer segmentation thicknesses between patients with or without a history of optic neuritis(ON)in NMOSD and the control group,including patients with MS,HC,and ION.Results:Forty-two studies were included and the interval between the last ON onset and examination was greater than 3 months.Compared with that in HC eyes,the loss of retinal nerve fiber layer(RNFL)and macular ganglion cell and inner plexiform layer(GC-IPL)was serious in NMOSD eye especially after ON.Moreover,compared with that in ION eyes or MS-related-ON eyes,the injury to the peripapillary retinal nerve fiber layer(pRNFL)was severe in NMOSD-related-ON eyes.In addition,the correlation coefficient between pRNFL and prognostic visual acuity was 0.43.However,the one-arm study revealed the inner nuclear layer(INL)was thickened in NMOSDrelated-ON eyes compared with HC eyes.Conclusions:Inclusion of the RNFL and macular GC-IPL is recommended for monitoring disease progression and attention should be paid to changes in the INL.
