Gamma-aminobutyric acid(GABA)ergic neurons,the most abundant inhibitory neurons in the human brain,have been found to be reduced in many neurological disorders,including Alzheimer's disease and Alzheimer's dis...Gamma-aminobutyric acid(GABA)ergic neurons,the most abundant inhibitory neurons in the human brain,have been found to be reduced in many neurological disorders,including Alzheimer's disease and Alzheimer's disease-related dementia.Our previous study identified the upregulation of microRNA-502-3p(miR-502-3p)and downregulation of GABA type A receptor subunitα-1 in Alzheimer's disease synapses.This study investigated a new molecular relationship between miR-502-3p and GABAergic synapse function.In vitro studies were perfo rmed using the mouse hippocampal neuronal cell line HT22 and miR-502-3p agomiRs and antagomiRs.In silico analysis identified multiple binding sites of miR-502-3p at GABA type A receptor subunitα-1 mRNA.Luciferase assay confirmed that miR-502-3p targets the GABA type A receptor subunitα-1 gene and suppresses the luciferase activity.Furthermore,quantitative reve rse transcription-polymerase chain reaction,miRNA in situ hybridization,immunoblotting,and immunostaining analysis confirmed that overexpression of miR-502-3p reduced the GABA type A receptor subunitα-1 level,while suppression of miR-502-3p increased the level of GABA type A receptor subunitα-1 protein.Notably,as a result of the overexpression of miR-502-3p,cell viability was found to be reduced,and the population of necrotic cells was found to be increased.The whole cell patch-clamp analysis of human-GABA receptor A-α1/β3/γ2L human embryonic kidney(HEK)recombinant cell line also showed that overexpression of miR-502-3p reduced the GABA current and overall GABA function,suggesting a negative correlation between miR-502-3p levels and GABAergic synapse function.Additionally,the levels of proteins associated with Alzheimer s disease were high with miR-502-3p overexpression and reduced with miR-502-3p suppression.The present study provides insight into the molecular mechanism of regulation of GABAergic synapses by miR-502-3p.We propose that micro-RNA,in particular miR-502-3p,could be a potential therapeutic to rget to modulate GABAergic synapse function in neurological disorders,including Alzheimer's disease and Alzheimer's diseaserelated dementia.展开更多
AIM:To observe the clinical efficacy of the combined use of small incision lenticule extraction(SMILE)-derived lenticule patches in corneal dermoid excision,with fixation of the lenticule patches assisted by fibrin gl...AIM:To observe the clinical efficacy of the combined use of small incision lenticule extraction(SMILE)-derived lenticule patches in corneal dermoid excision,with fixation of the lenticule patches assisted by fibrin glue.METHODS:Seventeen eyes of 17 patients with corneal dermoid were treated with dermoid removal combined with SMILE-derived lenticule transplantation.All lenticule patches were fixed by fibrin glue.Ocular changes were assessed using slit lamp microscopy and anterior-segmental optical coherence tomography.The best-corrected visual acuity(BCVA)and ocular dioptric variations were examined preoperatively and postoperatively.Intraocular pressure(IOP)was also monitored in all visited time.RESULTS:Totally,18 lenticule patches were used on 17 eyes of 17 cornea dermoid patients.The mean follow-up time was 11.47±5.28mo.All lenticule patches we resuccessfullyg lued,kept on its location and maintained transparent during the follow-up time,with a consecutive epithelial cover for 1wk.Nine of the patients could coordinate visual and optometry exam well.Their preoperative BCVA is 0.60±0.35 in decimal,significantly improved to 0.80±0.26 in decimal at 6mo postoperatively(Z=-2.392,P=0.017),but the changes of their corneal astigmatism diopters showed no significance,with 2.22±1.91 D preoperatively,and 2.28±1.31 D at 6mo postoperatively(Z=-0.135,P=0.893).Limbal pannus formation occurred in 4(23.52%)cases and decreased with the application of tacrolimus eyedrops.IOP increased in 2(11.76%)cases,but well decreased by timolol maleate eyedrops.All the adult patients or guardians of minor patients were satisfied with the cosmetic improvement.CONCLUSION:Dermoid excision combined with transplantation of SMILE-derived lenticule patches using fibrin glue is a safe and effective novel tectonic keratoplasty procedure for corneal dermoid.展开更多
In typical metal foundry applications,sand casting is still the most used technology.The related binder plays a very important role as its performances can directly influence the quality of castings.Among many binders...In typical metal foundry applications,sand casting is still the most used technology.The related binder plays a very important role as its performances can directly influence the quality of castings.Among many binders,glues of animal origin have attracted much attention in recent years due to their reduced environmental impact.How-ever,they display some drawbacks such as the tendency to coagulate easily at room temperature and a relatively low strength.In this study,a novel gas-hardening casting binder was prepared using an animal glue and anhy-drous potassium carbonate as a hydrolyzing agent to avoid undesired agglomeration.Moreover,sodium pyropho-sphate and furfuryl alcohol were exploited as modifiers to obtain a binder with a high compressive strength.The best modification conditions,determined by means of an orthogonal design matrix approach,were 4 g of Na2CO3,sodium pyrophosphate,furfuryl alcohol and animal glue with a ratio of 4:12:100,at 85°C and with a duration of 115 min,respectively.The viscosity of the mixture obtained under these optimized conditions was 1250 mPa⋅s.The compressive strength of the binder,hardened by CO_(2) gas,was 4.00 MPa.Its gas evolution at 850°C was 15 ml⋅g-1,and its residual strength after 10 min calculation at 800°C was 0.01 MPa,which is high enough to meet the requirement of core-making in foundry.Moreover,after hydrolysis and further modification,animal glue and modifiers displayed a grafting reaction and an esterification reaction,respectively,which made the adhesive network denser and improved its thermal stability.展开更多
Objective:CAR-T/NK cells have had limited success in the treatment of solid tumors,such as colorectal cancer(CRC),in part because of the heterogeneous nature of tumor-associated antigens that lead to antigen-negative ...Objective:CAR-T/NK cells have had limited success in the treatment of solid tumors,such as colorectal cancer(CRC),in part because of the heterogeneous nature of tumor-associated antigens that lead to antigen-negative relapse after the initial response.This barrier might be overcome by enhancing the recruitment and durability of endogenous immune cells.Methods:Immunohistochemistry and flow cytometry were used to assess the expression of CD133 antigen in tissue microarrays and cell lines,respectively.Retroviral vector transduction was used to generate CBLB502-secreting CAR133-NK92 cells(CAR133-i502-NK92).The tumor killing capacity of CAR133-NK92 cells in vitro and in vivo were quantified via LDH release,the RTCA assay,and the degranulation test,as well as measuring tumor bioluminescence signal intensity in mice xenografts.Results:We engineered CAR133-i502-NK92 cells and demonstrated that those cells displayed enhanced proliferation(9.0×10^(4)cells vs.7.0×10^(4)cells)and specific anti-tumor activities in vitro and in a xenogeneic mouse model,and were well-tolerated.Notably,CBLB502 secreted by CAR133-i502-NK92 cells effectively activated endogenous immune cells.Furthermore,in hCD133+/hCD133−mixed cancer xenograft models,CAR133-i502-NK92 cells suppressed cancer growth better than the counterparts(n=5,P=0.0297).Greater T-cell infiltration was associated with greater anti-tumor potency(P<0.0001).Conclusions:Armed with a CBLB502 TLR5 agonist,CAR133-NK92 cells were shown to be capable of specifically eliminating CD133-positive colon cancer cells in a CAR133-dependent manner and indirectly eradicating CD133-negative colon cancer cells in a CBLB502-specific endogenous immune response manner.This study describes a novel technique for optimizing CAR-T/NK cells for the treatment of antigenically-diverse solid tumors.展开更多
基金supported by the National Institute on Aging (NIA)National Institutes of Health (NIH)+3 种基金Nos.K99AG065645,R00AG065645R00AG065645-04S1 (to SK)NIH research grants,NINDS,No.R01 NS115834NINDS/NIA,No.R01 NS115834-02S1 (to LG)。
文摘Gamma-aminobutyric acid(GABA)ergic neurons,the most abundant inhibitory neurons in the human brain,have been found to be reduced in many neurological disorders,including Alzheimer's disease and Alzheimer's disease-related dementia.Our previous study identified the upregulation of microRNA-502-3p(miR-502-3p)and downregulation of GABA type A receptor subunitα-1 in Alzheimer's disease synapses.This study investigated a new molecular relationship between miR-502-3p and GABAergic synapse function.In vitro studies were perfo rmed using the mouse hippocampal neuronal cell line HT22 and miR-502-3p agomiRs and antagomiRs.In silico analysis identified multiple binding sites of miR-502-3p at GABA type A receptor subunitα-1 mRNA.Luciferase assay confirmed that miR-502-3p targets the GABA type A receptor subunitα-1 gene and suppresses the luciferase activity.Furthermore,quantitative reve rse transcription-polymerase chain reaction,miRNA in situ hybridization,immunoblotting,and immunostaining analysis confirmed that overexpression of miR-502-3p reduced the GABA type A receptor subunitα-1 level,while suppression of miR-502-3p increased the level of GABA type A receptor subunitα-1 protein.Notably,as a result of the overexpression of miR-502-3p,cell viability was found to be reduced,and the population of necrotic cells was found to be increased.The whole cell patch-clamp analysis of human-GABA receptor A-α1/β3/γ2L human embryonic kidney(HEK)recombinant cell line also showed that overexpression of miR-502-3p reduced the GABA current and overall GABA function,suggesting a negative correlation between miR-502-3p levels and GABAergic synapse function.Additionally,the levels of proteins associated with Alzheimer s disease were high with miR-502-3p overexpression and reduced with miR-502-3p suppression.The present study provides insight into the molecular mechanism of regulation of GABAergic synapses by miR-502-3p.We propose that micro-RNA,in particular miR-502-3p,could be a potential therapeutic to rget to modulate GABAergic synapse function in neurological disorders,including Alzheimer's disease and Alzheimer's diseaserelated dementia.
基金Supported by Science and Technology Planning Project of Guangxi Zhuang Autonomous Region (No.AB18221038)Guangxi Medical&Health Appropriate Technology Development and Promoted Application Project (No.S2021092)。
文摘AIM:To observe the clinical efficacy of the combined use of small incision lenticule extraction(SMILE)-derived lenticule patches in corneal dermoid excision,with fixation of the lenticule patches assisted by fibrin glue.METHODS:Seventeen eyes of 17 patients with corneal dermoid were treated with dermoid removal combined with SMILE-derived lenticule transplantation.All lenticule patches were fixed by fibrin glue.Ocular changes were assessed using slit lamp microscopy and anterior-segmental optical coherence tomography.The best-corrected visual acuity(BCVA)and ocular dioptric variations were examined preoperatively and postoperatively.Intraocular pressure(IOP)was also monitored in all visited time.RESULTS:Totally,18 lenticule patches were used on 17 eyes of 17 cornea dermoid patients.The mean follow-up time was 11.47±5.28mo.All lenticule patches we resuccessfullyg lued,kept on its location and maintained transparent during the follow-up time,with a consecutive epithelial cover for 1wk.Nine of the patients could coordinate visual and optometry exam well.Their preoperative BCVA is 0.60±0.35 in decimal,significantly improved to 0.80±0.26 in decimal at 6mo postoperatively(Z=-2.392,P=0.017),but the changes of their corneal astigmatism diopters showed no significance,with 2.22±1.91 D preoperatively,and 2.28±1.31 D at 6mo postoperatively(Z=-0.135,P=0.893).Limbal pannus formation occurred in 4(23.52%)cases and decreased with the application of tacrolimus eyedrops.IOP increased in 2(11.76%)cases,but well decreased by timolol maleate eyedrops.All the adult patients or guardians of minor patients were satisfied with the cosmetic improvement.CONCLUSION:Dermoid excision combined with transplantation of SMILE-derived lenticule patches using fibrin glue is a safe and effective novel tectonic keratoplasty procedure for corneal dermoid.
基金supported by the National Natural Science Foundation of China(51275313)Shandong Province Transportation Science and Technology Project(2021B115)Shandong Jiaotong University School Fund(Z2019036).
文摘In typical metal foundry applications,sand casting is still the most used technology.The related binder plays a very important role as its performances can directly influence the quality of castings.Among many binders,glues of animal origin have attracted much attention in recent years due to their reduced environmental impact.How-ever,they display some drawbacks such as the tendency to coagulate easily at room temperature and a relatively low strength.In this study,a novel gas-hardening casting binder was prepared using an animal glue and anhy-drous potassium carbonate as a hydrolyzing agent to avoid undesired agglomeration.Moreover,sodium pyropho-sphate and furfuryl alcohol were exploited as modifiers to obtain a binder with a high compressive strength.The best modification conditions,determined by means of an orthogonal design matrix approach,were 4 g of Na2CO3,sodium pyrophosphate,furfuryl alcohol and animal glue with a ratio of 4:12:100,at 85°C and with a duration of 115 min,respectively.The viscosity of the mixture obtained under these optimized conditions was 1250 mPa⋅s.The compressive strength of the binder,hardened by CO_(2) gas,was 4.00 MPa.Its gas evolution at 850°C was 15 ml⋅g-1,and its residual strength after 10 min calculation at 800°C was 0.01 MPa,which is high enough to meet the requirement of core-making in foundry.Moreover,after hydrolysis and further modification,animal glue and modifiers displayed a grafting reaction and an esterification reaction,respectively,which made the adhesive network denser and improved its thermal stability.
基金supported by the Technology Innovation and Application Developnent Key Program of Chongqing(Grant No.CSTC2021jscx-gksb-N0026)the National Natural Science Foundation of China(Grant No.31540016)+1 种基金the Basic Research and Frontier Exploration Projects of Chongqing(Grant No.cstc2018jcyjAX0075)the Subsidy Fund for the Development of National Silk in Chongqing(Grant No.CQ2018JSCE05).
文摘Objective:CAR-T/NK cells have had limited success in the treatment of solid tumors,such as colorectal cancer(CRC),in part because of the heterogeneous nature of tumor-associated antigens that lead to antigen-negative relapse after the initial response.This barrier might be overcome by enhancing the recruitment and durability of endogenous immune cells.Methods:Immunohistochemistry and flow cytometry were used to assess the expression of CD133 antigen in tissue microarrays and cell lines,respectively.Retroviral vector transduction was used to generate CBLB502-secreting CAR133-NK92 cells(CAR133-i502-NK92).The tumor killing capacity of CAR133-NK92 cells in vitro and in vivo were quantified via LDH release,the RTCA assay,and the degranulation test,as well as measuring tumor bioluminescence signal intensity in mice xenografts.Results:We engineered CAR133-i502-NK92 cells and demonstrated that those cells displayed enhanced proliferation(9.0×10^(4)cells vs.7.0×10^(4)cells)and specific anti-tumor activities in vitro and in a xenogeneic mouse model,and were well-tolerated.Notably,CBLB502 secreted by CAR133-i502-NK92 cells effectively activated endogenous immune cells.Furthermore,in hCD133+/hCD133−mixed cancer xenograft models,CAR133-i502-NK92 cells suppressed cancer growth better than the counterparts(n=5,P=0.0297).Greater T-cell infiltration was associated with greater anti-tumor potency(P<0.0001).Conclusions:Armed with a CBLB502 TLR5 agonist,CAR133-NK92 cells were shown to be capable of specifically eliminating CD133-positive colon cancer cells in a CAR133-dependent manner and indirectly eradicating CD133-negative colon cancer cells in a CBLB502-specific endogenous immune response manner.This study describes a novel technique for optimizing CAR-T/NK cells for the treatment of antigenically-diverse solid tumors.
