BACKGROUND： Neural stem cell （NSC） survival is closely associated with cell apoptosis in ischemic-hypoxic regions following transplantation. Numerous studies have revealed that X-box binding protein 1 （XBP1） is a...BACKGROUND： Neural stem cell （NSC） survival is closely associated with cell apoptosis in ischemic-hypoxic regions following transplantation. Numerous studies have revealed that X-box binding protein 1 （XBP1） is a transcription factor during endoplasmic reticulum unfolded protein response and is essential for cell survival, differentiation, and anti-apoptotic effects. OBJECTIVE： To determine the effects of the XBP1 gene on NSC proliferation and apoptosis under hypoxic conditions following XBP1 gene transfection into rat embryonic hippocampal NSCs using recombinant adenovirus vector. DESIGN, TIME AND SETTING： In vitro experiments were performed at the Laboratory of Cell Biology of Jilin University and Laboratory of Proteomics, Department of Neurology, Jilin University China from September 2008 to November 2009. MATERIALS： Recombinant adenovirus package XBP1 gene and Ad-XBPl-enhanced green fluorescent protein plasmid （Guangzhou Easywin BioMed Technology, China）, rabbit anti-XBP1 and its target gene estrogen receptor degradation-enhancing a-mannosidase-like protein （EDEM） glucose-regulated protein 78 （GRP78）, anti-apoptotic molecule Bcl-2 and proapoptotic molecule Bax polyclonal antibody （Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA）, and COCI2 （Sigma, St. Louis, MO, USA） were used in the present study. METHODS： Hippocampi from embryonic, Sprague Dawley rats on gestational day 16 were harvested for NSC isolation and cloning, followed by immunofluorescence for Nestin and sub-culturing. The recombinant adenovirus Ad-XBPl-enhanced green fluorescent protein plasmid was transfected into rat embryonic hippocampal NSCs, and then CoCl2 was applied to induce hypoxia. MAIN OUTCOME MEASURES： Cell quantification and 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide colorimetric assay were utilized to detect proliferation in XBPl-transfected NSCs for 7 consecutive days. Western blot assay was utilized to quantify XBP1 GRP78, EDEM, Bcl-2, and Bax expression. Flow cytometry was used to measure apoptosis. RESULTS： NSC proliferation was significantly enhanced following XBP1 gene transfection （P 〈 0.05）. Under hypoxic conditions, GRP78, EDEM, and Bcl-2 levels increased, but Bax levels decreased. In addition, NSC apoptosis decreased following transfection （P 〈 0.05）. CONCLUSION： The XBP1 gene was successfully transfected into rat embryonic hippocampal NSCs using a recombinant adenovirus vector. NSC proliferation following transfection, as well as anti-apoptotic effects under hypoxia, was significantly increased.展开更多
Recombinant human granulocyte-colony stimulating factor （hG-CSF） has been shown to protect the nervous system after brain ischemia. However, the neuroprotective mechanism of hG-CSF remains unclear. The present study...Recombinant human granulocyte-colony stimulating factor （hG-CSF） has been shown to protect the nervous system after brain ischemia. However, the neuroprotective mechanism of hG-CSF remains unclear. The present study established a rat model of cerebral ischemia/reperfusion and subcutaneously injected recombinant hG-CSF after reperfusion for 2 hours. Cerebral cortical protein was extracted following 14 days of reperfusion and subjected to two-dimensional electrophoresis. In brain ischemic rats, 56 different protein spots were screened, including 17 that were upregulated and 17 that were downregulated, compared with the sham-surgery group. Matrix assisted laser desorption ionization/time of flight mass spectrometry was used to determine peptide mass fingerprinting. Following a National Center for Biotechnology Information database search and confirmation with the Swiss-Prot database, 19 spots were identified as known proteins. Following hG-CSF treatment, 35 different protein spots were found, including 16 that were downregulated and 19 that were upregulated. Six were known proteins, including dihydropyrimidinase-associated protein 2, glial fibrillary acidic protein, endomucin, Rho GDP dissociation inhibitor, Rab GDP dissociation inhibitor and guanine-nucleotide-binding protein. Results indicate that hG-CSF is involved in neuroprotection after brain ischemia, possibly by regulating the expression of various neural regeneration-associated proteins at the subacute stage.展开更多
Bietti crystalline corneoretinal dystrophy is an inherited retinal disease caused by mutations in CYP4V2,which results in blindness in the working-age population,and there is currently no available treatment.Here,we r...Bietti crystalline corneoretinal dystrophy is an inherited retinal disease caused by mutations in CYP4V2,which results in blindness in the working-age population,and there is currently no available treatment.Here,we report the results of the first-in-human clinical trial(NCT04722107)of gene therapy for Bietti crystalline corneoretinal dystrophy,including 12 participants who were followed up for 180-365 days.This open-label,single-arm exploratory trial aimed to assess the safety and efficacy of a recombinant adeno-associated-virus-serotype-2/8 vector encoding the human CYP4V2 protein(rAAV2/8-hCYP4V2).Participants received a single unilateral subretinal injection of 7.5×10^(10)vector genomes of rAAV2/8-hCYP4V2.Overall,73 treatment-emergent adverse events were reported,with the majority(98.6%)being of mild or moderate intensity and considered to be procedure-or corticosteroid-related;no treatment-related serious adverse events or local/systemic immune toxicities were observed.Compared with that measured at baseline,77.8%of the treated eyes showed improvement in best-corrected visual acuity(BCVA)on day 180,with a mean±standard deviation increase of 9.0±10.8 letters in the 9 eyes analyzed(p=0.021).By day 365,80%of the treated eyes showed an increase in BCVA,with a mean increase of 11.0±10.6 letters in the 5 eyes assessed(p=0.125).Importantly,the patients’improvement observed using multifocal electroretinogram,microperimetry,and Visual Function Questionnaire-25 further supported the beneficial effects of the treatment.We conclude that the favorable safety profile and visual improvements identified in this trial encourage the continued development of rAAV2/8-hCYP4V2(named ZVS101e).展开更多
The shuttle effect hinders the practical application of lithium-sulfur(Li-S)batteries due to the poor affinity between a substrate and Li polysulfides(LiPSs)and the sluggish transition of soluble LiPSs to insoluble Li...The shuttle effect hinders the practical application of lithium-sulfur(Li-S)batteries due to the poor affinity between a substrate and Li polysulfides(LiPSs)and the sluggish transition of soluble LiPSs to insoluble Li2S or elemental S.Here,we report that Ni hexatomic clusters embedded in a nitrogen-doped three-dimensional(3D)graphene framework(Ni-N/G)possess stronger interaction with soluble polysulfides than that with insoluble polysulfides.The synthetic electrocatalyst deployed in the sulfur cathode plays a multifunctional role:(i)selectively adsorbing the polysulfides dissolved in the electrolyte,(ii)expediting the sluggish liquid-solid phase transformations at the active sites as electrocatalysts,and(iii)accelerating the kinetics of the electrochemical reaction of multielectron sulfur,thereby inhibiting the dissolution of LiPSs.The constructed S@Ni-N/G cathode delivers an areal capacity of 9.43mAhcm^(-2) at 0.1 C at S loading of 6.8 mg cm^(-2),and it exhibits a gravimetric capacity of 1104mAhg^(-1) with a capacity fading rate of 0.045%per cycle over 50 cycles at 0.2 C at S loading of 2.0 mg cm^(-2).This work opens a rational approach to achieve the selective adsorption and expediting of polysulfide transition for the performance enhancement of Li-S batteries.展开更多
文摘BACKGROUND： Neural stem cell （NSC） survival is closely associated with cell apoptosis in ischemic-hypoxic regions following transplantation. Numerous studies have revealed that X-box binding protein 1 （XBP1） is a transcription factor during endoplasmic reticulum unfolded protein response and is essential for cell survival, differentiation, and anti-apoptotic effects. OBJECTIVE： To determine the effects of the XBP1 gene on NSC proliferation and apoptosis under hypoxic conditions following XBP1 gene transfection into rat embryonic hippocampal NSCs using recombinant adenovirus vector. DESIGN, TIME AND SETTING： In vitro experiments were performed at the Laboratory of Cell Biology of Jilin University and Laboratory of Proteomics, Department of Neurology, Jilin University China from September 2008 to November 2009. MATERIALS： Recombinant adenovirus package XBP1 gene and Ad-XBPl-enhanced green fluorescent protein plasmid （Guangzhou Easywin BioMed Technology, China）, rabbit anti-XBP1 and its target gene estrogen receptor degradation-enhancing a-mannosidase-like protein （EDEM） glucose-regulated protein 78 （GRP78）, anti-apoptotic molecule Bcl-2 and proapoptotic molecule Bax polyclonal antibody （Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA）, and COCI2 （Sigma, St. Louis, MO, USA） were used in the present study. METHODS： Hippocampi from embryonic, Sprague Dawley rats on gestational day 16 were harvested for NSC isolation and cloning, followed by immunofluorescence for Nestin and sub-culturing. The recombinant adenovirus Ad-XBPl-enhanced green fluorescent protein plasmid was transfected into rat embryonic hippocampal NSCs, and then CoCl2 was applied to induce hypoxia. MAIN OUTCOME MEASURES： Cell quantification and 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide colorimetric assay were utilized to detect proliferation in XBPl-transfected NSCs for 7 consecutive days. Western blot assay was utilized to quantify XBP1 GRP78, EDEM, Bcl-2, and Bax expression. Flow cytometry was used to measure apoptosis. RESULTS： NSC proliferation was significantly enhanced following XBP1 gene transfection （P 〈 0.05）. Under hypoxic conditions, GRP78, EDEM, and Bcl-2 levels increased, but Bax levels decreased. In addition, NSC apoptosis decreased following transfection （P 〈 0.05）. CONCLUSION： The XBP1 gene was successfully transfected into rat embryonic hippocampal NSCs using a recombinant adenovirus vector. NSC proliferation following transfection, as well as anti-apoptotic effects under hypoxia, was significantly increased.
文摘Recombinant human granulocyte-colony stimulating factor （hG-CSF） has been shown to protect the nervous system after brain ischemia. However, the neuroprotective mechanism of hG-CSF remains unclear. The present study established a rat model of cerebral ischemia/reperfusion and subcutaneously injected recombinant hG-CSF after reperfusion for 2 hours. Cerebral cortical protein was extracted following 14 days of reperfusion and subjected to two-dimensional electrophoresis. In brain ischemic rats, 56 different protein spots were screened, including 17 that were upregulated and 17 that were downregulated, compared with the sham-surgery group. Matrix assisted laser desorption ionization/time of flight mass spectrometry was used to determine peptide mass fingerprinting. Following a National Center for Biotechnology Information database search and confirmation with the Swiss-Prot database, 19 spots were identified as known proteins. Following hG-CSF treatment, 35 different protein spots were found, including 16 that were downregulated and 19 that were upregulated. Six were known proteins, including dihydropyrimidinase-associated protein 2, glial fibrillary acidic protein, endomucin, Rho GDP dissociation inhibitor, Rab GDP dissociation inhibitor and guanine-nucleotide-binding protein. Results indicate that hG-CSF is involved in neuroprotection after brain ischemia, possibly by regulating the expression of various neural regeneration-associated proteins at the subacute stage.
基金This work was supported by the National Key Research and Development Program during the 14th Five-year Plan Period(2023YFC2706304)the National Key R&D Program of China(2023YFC3403300)+4 种基金National Natural Science Foundation of China(82220108017,82141128,82371074)Capital Health Development Research Special Project(2024-1-2052)Beijing Science and Technology Program(Z231100004823021,Z201100005520045)Beijing Natural Science Foundation(J230031)Sanming Project of Medicine in Shenzhen(No.SZSM202311018)and Chigenovo Co.,Ltd.
文摘Bietti crystalline corneoretinal dystrophy is an inherited retinal disease caused by mutations in CYP4V2,which results in blindness in the working-age population,and there is currently no available treatment.Here,we report the results of the first-in-human clinical trial(NCT04722107)of gene therapy for Bietti crystalline corneoretinal dystrophy,including 12 participants who were followed up for 180-365 days.This open-label,single-arm exploratory trial aimed to assess the safety and efficacy of a recombinant adeno-associated-virus-serotype-2/8 vector encoding the human CYP4V2 protein(rAAV2/8-hCYP4V2).Participants received a single unilateral subretinal injection of 7.5×10^(10)vector genomes of rAAV2/8-hCYP4V2.Overall,73 treatment-emergent adverse events were reported,with the majority(98.6%)being of mild or moderate intensity and considered to be procedure-or corticosteroid-related;no treatment-related serious adverse events or local/systemic immune toxicities were observed.Compared with that measured at baseline,77.8%of the treated eyes showed improvement in best-corrected visual acuity(BCVA)on day 180,with a mean±standard deviation increase of 9.0±10.8 letters in the 9 eyes analyzed(p=0.021).By day 365,80%of the treated eyes showed an increase in BCVA,with a mean increase of 11.0±10.6 letters in the 5 eyes assessed(p=0.125).Importantly,the patients’improvement observed using multifocal electroretinogram,microperimetry,and Visual Function Questionnaire-25 further supported the beneficial effects of the treatment.We conclude that the favorable safety profile and visual improvements identified in this trial encourage the continued development of rAAV2/8-hCYP4V2(named ZVS101e).
基金This work was supported by the National Key R&D Program of China(2018YFB0104300,2016YFA0200102)National Natural Science Foundation of China(51874104,51631001)Key Technology and Supporting Platform of Genetic Engineering of Materials under State’s Key Project of Research and Development Plan(2016YFB0700600).
文摘The shuttle effect hinders the practical application of lithium-sulfur(Li-S)batteries due to the poor affinity between a substrate and Li polysulfides(LiPSs)and the sluggish transition of soluble LiPSs to insoluble Li2S or elemental S.Here,we report that Ni hexatomic clusters embedded in a nitrogen-doped three-dimensional(3D)graphene framework(Ni-N/G)possess stronger interaction with soluble polysulfides than that with insoluble polysulfides.The synthetic electrocatalyst deployed in the sulfur cathode plays a multifunctional role:(i)selectively adsorbing the polysulfides dissolved in the electrolyte,(ii)expediting the sluggish liquid-solid phase transformations at the active sites as electrocatalysts,and(iii)accelerating the kinetics of the electrochemical reaction of multielectron sulfur,thereby inhibiting the dissolution of LiPSs.The constructed S@Ni-N/G cathode delivers an areal capacity of 9.43mAhcm^(-2) at 0.1 C at S loading of 6.8 mg cm^(-2),and it exhibits a gravimetric capacity of 1104mAhg^(-1) with a capacity fading rate of 0.045%per cycle over 50 cycles at 0.2 C at S loading of 2.0 mg cm^(-2).This work opens a rational approach to achieve the selective adsorption and expediting of polysulfide transition for the performance enhancement of Li-S batteries.
