●AIM:To explore whether autophagy functions as a cellular adaptation mechanism in lens epithelial cells(LECs)under hyperosmotic stress.●METHODS:LECs were treated with hyperosmotic stress at the concentration of 270,...●AIM:To explore whether autophagy functions as a cellular adaptation mechanism in lens epithelial cells(LECs)under hyperosmotic stress.●METHODS:LECs were treated with hyperosmotic stress at the concentration of 270,300,400,500,or 600 mOsm for 6,12,18,24h in vitro.Polymerase chain reaction(PCR)was employed for the mRNA expression of autophagyrelated genes,while Western blotting detected the targeted protein expression.The transfection of stub-RFP-sens-GFPLC3 autophagy-related double fluorescence lentivirus was conducted to detect the level of autophagy flux.Scanning electron microscopy was used to detect the existence of autolysosome.Short interfering RNA of autophagy-related gene(ATG)7,transient receptor potential vanilloid(TRPV)1 overexpression plasmid,related agonists and inhibitors were employed to their influence on autophagy related pathway.Flow cytometry was employed to test the apoptosis and intracellular Ca^(2+)level.Mitochondrial membrane potential was measured by JC-1 staining.The cell counting kit-8 assay was used to calculate the cellular viability.The wound healing assay was used to evaluate the wound closure rate.GraphPad 6.0 software was utilized to evaluate the data.●RESULTS:The hyperosmotic stress activated autophagy in a pressure-and time-dependent manner in LECs.Beclin 1 protein expression and conversion of LC3B II to LC3B I increased,whereas sequestosome-1(SQSTM1)protein expression decreased.Transient Ca^(2+)influx was stimulated caused by hyperosmotic stress,levels of mammalian target of rapamycin(mTOR)phosphorylation decreased,and the level of AMP-activated protein kinase(AMPK)phosphorylation increased in the early stage.Based on this evidence,autophagy activation through the Ca^(2+)-dependent AMPK/mTOR pathway might represent an adaptation process in LECs under hyperosmotic stress.Hyperosmotic stress decreased cellular viability and accelerated apoptosis in LECs and cellular migration decreased.Inhibition of autophagy by ATG7 knockdown had similar results.TRPV1 overexpression increased autophagy and might be crucial in the occurrence of autophagy promoted by hyperosmotic stress.●CONCLUSION:A combination of hyperosmotic stress and autophagy inhibition may be a promising approach to decrease the number of LECs in the capsular bag and pave the way for improving prevention of posterior capsular opacification and capsular fibrosis.展开更多
AIM:To characterize the N6-methyladenosine(m6A)modification patterns in long non-coding RNAs(lncRNAs)in sporadic congenital cataract(CC)and age-related cataract(ARC).METHODS:Anterior capsule of the lens were collected...AIM:To characterize the N6-methyladenosine(m6A)modification patterns in long non-coding RNAs(lncRNAs)in sporadic congenital cataract(CC)and age-related cataract(ARC).METHODS:Anterior capsule of the lens were collected from patients with CC and ARC.Methylated RNA immunoprecipitation with next-generation sequencing and RNA sequencing were performed to identify m6A-tagged lncRNAs and lncRNAs expression.Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses and Gene Ontology annotation were used to predict potential functions of the m6A-lncRNAs.RESULTS:Large amount of m6A peaks within lncRNA were identified for both CC and ARC,while the level was much higher in ARC(49870 peaks)than that in CC(18688 peaks),yet those difference between ARC in younger age group(ARC-1)and ARC in elder age group(ARC-2)was quite slight.A total of 1305 hypermethylated and 1178 hypomethylated lncRNAs,as well as 182 differential expressed lncRNAs were exhibited in ARC compared with CC.On the other hand,5893 hypermethylated and 5213 hypomethylated lncRNAs,as well as 155 significantly altered lncRNA were identified in ARC-2 compared with ARC-1.Altered lncRNAs in ARC were mainly associated with the organization and biogenesis of intracellular organelles,as well as nucleotide excision repair.CONCLUSION:Our results for the first time present an overview of the m6A methylomes of lncRNA in CC and ARC,providing a solid basis and uncovering a new insight to reveal the potential pathogenic mechanism of CC and ARC.展开更多
AIM:To demonstrate local dry vitrectomy combined with segmental scleral buckling and viscoelastic tamponade for the treatment of partial rhegmatogenous retinal detachment(RRD)with local vitreous traction in patients a...AIM:To demonstrate local dry vitrectomy combined with segmental scleral buckling and viscoelastic tamponade for the treatment of partial rhegmatogenous retinal detachment(RRD)with local vitreous traction in patients at high-risk for proliferative vitreoretinopathy(PVR).METHODS:Eleven eyes of 11 patients were retrospectively studied,including 5 retinal dialysis and 6 retinal detachment(RD;5 eyes with peripheral retinal hole and I eye with giant tear).All patients exhibited partial RD and local vitreous traction.Combined local dry vitrectomy without conventional infusion and segmental scleral buckling was performed.Viscoelastic fluid was injected into the vitreous cavity if needed Demographic information,preoperative and postoperative complications,and outcomes were recorded.RESULTS:The mean age of the patients at presentation was 26.55±13.52y.All 11 patients obtained retinal reattachment after a single surgical intervention.Postoperative visual acuities were improved or remained stable in all patients.None of them developed complications,except for temporary mildly increased intraocular pressure in 3 cases.CONCLUSION:Combined local dry vitrectomy and segmental scleral buckling are effective for patients of RRD with local vitreous traction.The technique avoids many complications associated with regular surgery and was minimally invasive to both the external and internal eye.展开更多
·AIM:To verify the feasibility and safety of staged lensectomy and vitrectomy in stage 5 C retinopathy of prematurity(ROP)with corneal opacification.·METHODS:This was a retrospective,interventional,consecuti...·AIM:To verify the feasibility and safety of staged lensectomy and vitrectomy in stage 5 C retinopathy of prematurity(ROP)with corneal opacification.·METHODS:This was a retrospective,interventional,consecutive case series.Twenty-two eyes of 18 stage 5 C ROP patients with corneal opacification were included.Regular combined lensectomy and vitrectomy were not prescribed due to the invisible fundus.Staged lensectomy and posterior vitrectomy were performed.The anatomical and visual outcomes were reviewed at the final follow-up visit.·RESULTS:The mean gestational age of ROP patients was29.3±1.6 wk(range:27-32 wk),comprising 8 males and 10 females.The average birth weight was 1363.0±300.0 g.All the eyes had corneal opacity and flat or disappeared anterior chambers pre-operatively.Two eyes had complicated cataract and 7 eyes had retrolental fibroplasia.Six eyes had posterior pupillary synechiae or membranes.Seven(31.8%)eyes had vascularly active retinas.The average interval between two procedures was 6.8±4.6 mo(2.5-18.5 mo).After surgeries,all the patients had normal anterior chambers.Fourteen eyes had clear corneas.The intraocular pressure of 3 eyes with glaucoma was controlled by medication.Two eyes had ocular phthisis.The retina was reattached in 3 eyes and partially attached in 11 eyes.Visual acuity ranged from no light perception to hand motion.·CONCLUSION:Staged lensectomy and vitrectomy are procedures that can halt progression to further complications and preserve some useful eyesight in stage5 C ROP patients with corneal opacification.The earlier the lensectomy is performed,the better the prognosis is.展开更多
文摘●AIM:To explore whether autophagy functions as a cellular adaptation mechanism in lens epithelial cells(LECs)under hyperosmotic stress.●METHODS:LECs were treated with hyperosmotic stress at the concentration of 270,300,400,500,or 600 mOsm for 6,12,18,24h in vitro.Polymerase chain reaction(PCR)was employed for the mRNA expression of autophagyrelated genes,while Western blotting detected the targeted protein expression.The transfection of stub-RFP-sens-GFPLC3 autophagy-related double fluorescence lentivirus was conducted to detect the level of autophagy flux.Scanning electron microscopy was used to detect the existence of autolysosome.Short interfering RNA of autophagy-related gene(ATG)7,transient receptor potential vanilloid(TRPV)1 overexpression plasmid,related agonists and inhibitors were employed to their influence on autophagy related pathway.Flow cytometry was employed to test the apoptosis and intracellular Ca^(2+)level.Mitochondrial membrane potential was measured by JC-1 staining.The cell counting kit-8 assay was used to calculate the cellular viability.The wound healing assay was used to evaluate the wound closure rate.GraphPad 6.0 software was utilized to evaluate the data.●RESULTS:The hyperosmotic stress activated autophagy in a pressure-and time-dependent manner in LECs.Beclin 1 protein expression and conversion of LC3B II to LC3B I increased,whereas sequestosome-1(SQSTM1)protein expression decreased.Transient Ca^(2+)influx was stimulated caused by hyperosmotic stress,levels of mammalian target of rapamycin(mTOR)phosphorylation decreased,and the level of AMP-activated protein kinase(AMPK)phosphorylation increased in the early stage.Based on this evidence,autophagy activation through the Ca^(2+)-dependent AMPK/mTOR pathway might represent an adaptation process in LECs under hyperosmotic stress.Hyperosmotic stress decreased cellular viability and accelerated apoptosis in LECs and cellular migration decreased.Inhibition of autophagy by ATG7 knockdown had similar results.TRPV1 overexpression increased autophagy and might be crucial in the occurrence of autophagy promoted by hyperosmotic stress.●CONCLUSION:A combination of hyperosmotic stress and autophagy inhibition may be a promising approach to decrease the number of LECs in the capsular bag and pave the way for improving prevention of posterior capsular opacification and capsular fibrosis.
基金Supported by the National Natural Science Foundation of China(No.82171069No.82371070)+3 种基金Shanghai Science and Technology Committee(No.22015820200)Shanghai Municipal Health Commission Innovative Medical Device Application Demonstration Project(No.23SHS03500-03)Project of Shanghai Municipal Commission of Health and Family Planning(No.202140224)Grants from Interdisciplinary Program of Shanghai Jiao Tong University(No.YG2021QN52).
文摘AIM:To characterize the N6-methyladenosine(m6A)modification patterns in long non-coding RNAs(lncRNAs)in sporadic congenital cataract(CC)and age-related cataract(ARC).METHODS:Anterior capsule of the lens were collected from patients with CC and ARC.Methylated RNA immunoprecipitation with next-generation sequencing and RNA sequencing were performed to identify m6A-tagged lncRNAs and lncRNAs expression.Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses and Gene Ontology annotation were used to predict potential functions of the m6A-lncRNAs.RESULTS:Large amount of m6A peaks within lncRNA were identified for both CC and ARC,while the level was much higher in ARC(49870 peaks)than that in CC(18688 peaks),yet those difference between ARC in younger age group(ARC-1)and ARC in elder age group(ARC-2)was quite slight.A total of 1305 hypermethylated and 1178 hypomethylated lncRNAs,as well as 182 differential expressed lncRNAs were exhibited in ARC compared with CC.On the other hand,5893 hypermethylated and 5213 hypomethylated lncRNAs,as well as 155 significantly altered lncRNA were identified in ARC-2 compared with ARC-1.Altered lncRNAs in ARC were mainly associated with the organization and biogenesis of intracellular organelles,as well as nucleotide excision repair.CONCLUSION:Our results for the first time present an overview of the m6A methylomes of lncRNA in CC and ARC,providing a solid basis and uncovering a new insight to reveal the potential pathogenic mechanism of CC and ARC.
基金Supported by the National Natural Science Foundation of China(No.81770963,No.81500725,No.81770964)Shanghai Shen Kang Hospital Development Center Program(No.16CR4017A).
文摘AIM:To demonstrate local dry vitrectomy combined with segmental scleral buckling and viscoelastic tamponade for the treatment of partial rhegmatogenous retinal detachment(RRD)with local vitreous traction in patients at high-risk for proliferative vitreoretinopathy(PVR).METHODS:Eleven eyes of 11 patients were retrospectively studied,including 5 retinal dialysis and 6 retinal detachment(RD;5 eyes with peripheral retinal hole and I eye with giant tear).All patients exhibited partial RD and local vitreous traction.Combined local dry vitrectomy without conventional infusion and segmental scleral buckling was performed.Viscoelastic fluid was injected into the vitreous cavity if needed Demographic information,preoperative and postoperative complications,and outcomes were recorded.RESULTS:The mean age of the patients at presentation was 26.55±13.52y.All 11 patients obtained retinal reattachment after a single surgical intervention.Postoperative visual acuities were improved or remained stable in all patients.None of them developed complications,except for temporary mildly increased intraocular pressure in 3 cases.CONCLUSION:Combined local dry vitrectomy and segmental scleral buckling are effective for patients of RRD with local vitreous traction.The technique avoids many complications associated with regular surgery and was minimally invasive to both the external and internal eye.
基金Supported by the National Natural Science Foundation of China(No.81770963No.81770964)。
文摘·AIM:To verify the feasibility and safety of staged lensectomy and vitrectomy in stage 5 C retinopathy of prematurity(ROP)with corneal opacification.·METHODS:This was a retrospective,interventional,consecutive case series.Twenty-two eyes of 18 stage 5 C ROP patients with corneal opacification were included.Regular combined lensectomy and vitrectomy were not prescribed due to the invisible fundus.Staged lensectomy and posterior vitrectomy were performed.The anatomical and visual outcomes were reviewed at the final follow-up visit.·RESULTS:The mean gestational age of ROP patients was29.3±1.6 wk(range:27-32 wk),comprising 8 males and 10 females.The average birth weight was 1363.0±300.0 g.All the eyes had corneal opacity and flat or disappeared anterior chambers pre-operatively.Two eyes had complicated cataract and 7 eyes had retrolental fibroplasia.Six eyes had posterior pupillary synechiae or membranes.Seven(31.8%)eyes had vascularly active retinas.The average interval between two procedures was 6.8±4.6 mo(2.5-18.5 mo).After surgeries,all the patients had normal anterior chambers.Fourteen eyes had clear corneas.The intraocular pressure of 3 eyes with glaucoma was controlled by medication.Two eyes had ocular phthisis.The retina was reattached in 3 eyes and partially attached in 11 eyes.Visual acuity ranged from no light perception to hand motion.·CONCLUSION:Staged lensectomy and vitrectomy are procedures that can halt progression to further complications and preserve some useful eyesight in stage5 C ROP patients with corneal opacification.The earlier the lensectomy is performed,the better the prognosis is.
