Postoperative complications of phacoemulsification,such as corneal edema caused by human corneal endothelial cell(CEC)injury,are still a matter of concern.Although several factors are known to cause CEC damage,the inf...Postoperative complications of phacoemulsification,such as corneal edema caused by human corneal endothelial cell(CEC)injury,are still a matter of concern.Although several factors are known to cause CEC damage,the influence of ultrasound on the formation of free radicals during surgery should be considered.Ultrasound in aqueous humor induces cavitation and promotes the formation of hydroxyl radicals or reactive oxygen species(ROS).ROS-induced apoptosis and autophagy in phacoemulsification have been suggested to significantly promote CEC injury.CEC cannot regenerate after injury,and measures must be taken to prevent the loss of CEC after phacoemulsification or other CEC injuries.Antioxidants can reduce the oxidative stress injury of CEC during phacoemulsification.Evidence from rabbit eye studies shows that ascorbic acid infusion during operation or local application of ascorbic acid during phacoemulsification has a protective effect by scavenging free radicals or reducing oxidative stress.Both in experiments and clinical practice,hydrogen dissolved in the irrigating solution can also prevent CEC damage during phacoemulsification surgery.Astaxanthin(AST)can inhibit oxidative damage,thereby protecting different cells from most pathological conditions,such as myocardial cells,luteinized granulosa cells of the ovary,umbilical vascular endothelial cells,and human retina pigment epithelium cell line(ARPE-19).However,existing research has not focused on the application of AST to prevent oxidative stress during phacoemulsification,and the related mechanisms need to be studied.The Rho related helical coil kinase inhibitor Y-27632 can inhibit CEC apoptosis after phacoemulsification.Rigorous experiments are required to confirm whether its effect is realized through improving the ROS clearance ability of CEC.展开更多
AIM:To compare the corneal outcome in Fuchs’endothelial dystrophy(FED)patients between femtosecond laser-assisted cataract surgery(FLACS)and conventional phaco surgery(CPS).METHODS:This was a randomized controlled st...AIM:To compare the corneal outcome in Fuchs’endothelial dystrophy(FED)patients between femtosecond laser-assisted cataract surgery(FLACS)and conventional phaco surgery(CPS).METHODS:This was a randomized controlled study comparing one eye surgery by FLACS and the contralateral eye operated by CPS(stop and chop technique)in FED patients.Central corneal thickness,corneal light backscatter,corneal densitometry,and central corneal endothelial cell count and hexagonality(noncontact endothelial cell microscope),and corrected distance visual acuity(CDVA)were assessed preoperatively and at day 1,40,and 180 postoperatively.RESULTS:Totally 31 patients(16 women)were included.At day 40 postoperatively,the mean endothelial cell loss(ECL)was 23.67%by FLACS and 17.30%by CPS(P=0.53).At day 180 postoperatively,ECL was 25.58%in FLACS and 21.32%in CPS(P=0.69).Densitometry data in all layers and all annuli from anterior layer to posterior layer in annuli 0-2,2-6,6-10 and 10-12,total densitometry with all layers and all annuli was performed.A significant difference was found in 6-10(posterior layer)at day 1 with-1.42 grayscale units(GSU;95%CI:-2.66 to-0.19,P=0.02).In 10-12(anterior layer,central layer and all layers)at day 40 were significant different with 7.7(95%CI:1.89 to 13.50,P=0.009),3.97(95%CI:0.23 to 7.71,P=0.03),4.73 GSU(95%CI:0.71 to 8.75,P=0.02),respectively.In the remaining parameters we found no difference between the two groups(P>0.05).Three CPS eyes suffered from corneal decompensation.CONCLUSION:There is no significant difference in corneal outcome between FLACS and CPS.Endothelial cell density and pentacam corneal outcome may be inadequate as outcome parameters in FED patients.展开更多
AIM:To transfect the cat corneal endothelial cells(CECs) with recombinant human β-nerve growth factor gene adeno-associated virus(AAV-β-NGF) and to observe the effect of the expressed β-NGF protein on the prolifera...AIM:To transfect the cat corneal endothelial cells(CECs) with recombinant human β-nerve growth factor gene adeno-associated virus(AAV-β-NGF) and to observe the effect of the expressed β-NGF protein on the proliferation activity of cat CECs.METHODS:The endothelium of cat cornea was torn under the microscope and rapidly cultivated in Dulbecco's modified Eagle's medium(DMEM) to form single layer CECs and the passage 2 endothelial cells were used in this experiment.The recombinant human AAV-β-NGF was constructed.The recombinant human AAV-β-NGF was transferred into cat CECs directly.Three groups were as following:normal CEC control group,CEC-AAV control group and recombinant CECAAV-β-NGF group.Forty-eight hours after transfection,the total RNA was extracted from the CEC by Trizol.The expression of the β-NGF target gene detected by fluorescence quantitative polymerase chain reaction;proliferation activity of the transfected CEC detected at 48 h by MTT assay;the percentage of G1 cells among CECs after transfect was detected by flow cytometry method(FCM);cell morphology was observed under inverted phase contrast microscope.RESULTS:The torn endothelium culture technique rapidly cultivated single layer cat corneal endothelial cells.The self-designed primers for the target gene and reference gene were efficient and special confirmed through electrophoresis analysis and DNA sequencing.Forty-eight hours after transfect,the human β-NGF gene mRNA detected by fluorescence quantitative polymerase chain reaction showed that there was no significant difference between normal CEC control group and CECAAV control group(P>0.05);there was significant difference between two control groups and recombinant CEC-AAV-β-NGF group(P<0.05).MTT assay showed that transfect of recombinant AAV-β-NGF promoted the proliferation activity of cat CEC,while there was no significant difference between normal CEC control group and CEC-AAV control group(P>0.05).FCM result showed that the percentage of G1 cells in CECAAV-NGF group was 76.8% while that in normal CEC control group and CEC-AAV control group was 46.6% and 49.8%.CONCLUSION:Recombinant AAV-β-NGF promotes proliferation in cat CECs by expressing bioactive β-NGF protein in high efficiency and suggests that its modulation can be used to treat vision loss secondary to corneal endotheiiai dysfunction.展开更多
Dear Sir,Iam Dr.Jaya Kaushik from the Department of Ophthalmology of the Post Graduate Institute of Medical Education and Research,Chandigarh,India.I write to present a case report of phacoemulsification in a rare cas...Dear Sir,Iam Dr.Jaya Kaushik from the Department of Ophthalmology of the Post Graduate Institute of Medical Education and Research,Chandigarh,India.I write to present a case report of phacoemulsification in a rare case of cataract associated with keratoconus and Fuch’s endothelial展开更多
AIM:To investigate the effects of a selective inhibitor of Rho-associated kinase(ROCK),Y-27632,on inbred Wuzhishan porcine corneal endothelial cells(PCECs)in vitro and in vivo studies.METHODS:Primary PCECs were trypsi...AIM:To investigate the effects of a selective inhibitor of Rho-associated kinase(ROCK),Y-27632,on inbred Wuzhishan porcine corneal endothelial cells(PCECs)in vitro and in vivo studies.METHODS:Primary PCECs were trypsinized from Wuzhishan miniature porcine corneal tissues.The optimal concentration of Y-27632 on PCECs was determined through MTT and 5-ethynyl-2'-deoxyuridine(EdU)-labeling assays.Seven New Zealand rabbits were used as a corneal endothelial dysfunction model,and a PCECs suspension supplemented with Y-27632 was injected into the anterior chamber of the rabbits.The progression of rabbit corneal opacity and edema were observed by slit lamp examination.The rabbits were sacrificed,and rabbit globes were enucleated for trypan blue-alizarin red staining,hematoxylineosin staining,and immunofluorescence analysis.RESULTS:Administration of 100μmol/L Y-27632 facilitated PCECs'proliferation obviously.The rabbit corneas injected with PCECs suspension and 100μmol/L Y-27632 were restored to transparency significantly after 14d.CONCLUSION:The 100μmol/L Y-27632 treatment improves PCECs'proliferation significantly.And our results suggest that Y-27632 and PCECs can be used to treat corneal endothelial dysfunction.展开更多
Background:The goal of this project was to analyze the relationship between cell morphology and proteases/proteases inhibitors(PIs)secretion profile in fuchs endothelial corneal dystrophy(FECD)corneal endothelial cell...Background:The goal of this project was to analyze the relationship between cell morphology and proteases/proteases inhibitors(PIs)secretion profile in fuchs endothelial corneal dystrophy(FECD)corneal endothelial cells(CECs).Methods:Cell morphology was determined using a circularity index(4π×area/perimeter2)for each CECs population extracted from surgical FECD specimens(N=2)and healthy Eye bank corneas(N=3).CECs were cultured 28 days post-confluency.Supernatant was collected and analysed using Proteome Profiler Array detecting 35 proteases and 32 PIs(R&D Systems).Proteome signal was analyzed using Image Studio Lite and correlated with the population’s circularity index.Results:Calculation of circularity index reported different morphologies among FECD populations(0.59±0.18 and 0.64±0.17)and healthy populations(0.44±0.18,0.66±0.13 and 0.71±0.11).Proteome arrays revealed the presence of 10 proteases(ADAMTS1,Cathepsin A,B,D,and X/Z/P,DPPIV/CD26,MMP-2,3 and 12,uPA/Urokinase)and 10 PIs(Protease Nexin II,Cystatin B and C,EMMPRIN/CD147,Latexin,Lipocalin-1,Serpin E1,TFPI,TFPI-2,TIMP-1,2 and 4).Healthy and FECD specimens showed similar variation patterns according to morphology for secretion of ADAMTS1,MMP-3 and 12.However,opposing patterns between healthy and FECD populations were observed for Cathepsin B and D.Moreover,some proteins did not show variation according to phenotype in healthy CECs,but did in FECD CECs:Cathepsin A,Cystatin C,TFPI-2 and total TIMPs.For the other proteins,secretion did not vary according to morphology or no specific pattern was distinguishable.Conclusions:To conclude,our results suggest that cell phenotype is linked to the secretion of certain proteases/PIs in both groups.However,there seems to be differences in secretion of particular proteases and PIs between FECD and healthy specimens as morphology did not have a similar influence.These differences might initiate an imbalance between proteases and PIs explaining the irregular thickening of the Descemet membrane seen in FECD.展开更多
Three months after surgery, the research group showed significantly statistical improvement in visual acuity, a statistically significant decrease in corneal endothelial cell density, a statistically significant incre...Three months after surgery, the research group showed significantly statistical improvement in visual acuity, a statistically significant decrease in corneal endothelial cell density, a statistically significant increase in the percentage of 5 and 8 sided cells and a statistically significant decrease in the percentage of six sided cells. Central corneal thickness and the percentage of 4 and 7 and more than 8 sided did not change in a statistically significant way. Comparing the test group and control group, no statistically significant differences were detected in the examined parameters. The present study also shows that the cornea in the eyes with congenital cataract does not show statistically significant changes in the density and the morphology of the corneal endothelial cells and the thickness of the cornea and in terms of corneal thickness in comparison to the corneas of healthy eyes. Although in corneas undergoing cataract occurs statistically significant changes, the influence of the cornea does not affect the improvement in visual acuity which was also demonstrated in this study.展开更多
AIM:To investigate the safety and efficacy of using a onestep viscoelastic agent technique for posterior chamber phakic implantable collamer lens with a central hole(ICL V4 c)implantation for myopia correction.METHODS...AIM:To investigate the safety and efficacy of using a onestep viscoelastic agent technique for posterior chamber phakic implantable collamer lens with a central hole(ICL V4 c)implantation for myopia correction.METHODS:The one-step viscoelastic agent technique for ICL V4 c implantation was used in 100 eyes of 52 patients.Refractive outcomes,intraocular pressure(IOP),corneal endothelial cell,and corneal densitometry values were evaluated at 1 d,1 wk,1 and 3 mo postoperatively.RESULTS:All the surgeries were uneventful.No corrected distance visual acuity was lost after 3 mo.IOP was 16.12±3.18 mm Hg before surgery,and 14.74±3.08 mm Hg at 1d and 14.50±2.56 mm Hg at 3 mo after surgery(P<0.05).Corneal endothelial cell density was 2580±242 cell/mm^(2),the coefficient of variation in cell size was 42.11%±7.92%,and the percentage of hexagonal cells was 40.98%±9.46%before surgery.No significant difference was found when these outcomes were compared between the studied time points(P>0.05).The corneal densitometry values of the central 2 mm and 2 to 6 mm areas showed similar regularities.After surgery,the values significantly increased at 1 d,then decreased to the preoperative values at 1 wk,and then continued to decrease at 3 mo(P<0.05).CONCLUSION:The one-step viscoelastic agent technique for ICL V4c implantation is found to be safe and effective for myopia correction and causes little disturbance to the cornea.展开更多
AIM: To compare the corneal endothelial cell counts pre-and post-operation, ultrastructure of anterior lens capsule and surgical completion after diathermic highfrequency capsulorhexis(DHC) and continuous curvilinear ...AIM: To compare the corneal endothelial cell counts pre-and post-operation, ultrastructure of anterior lens capsule and surgical completion after diathermic highfrequency capsulorhexis(DHC) and continuous curvilinear capsulorrhexis(CCC) applied in phacoemulsification(PHACO) of white cataract surgery.METHODS: Sixty-six eyes of 66 patients(33 males and 33 females) with cataract aged between 60 and 80 y(mean 72.5±5.5) were recruited and undergone the surgery from June 2014 to November 2016. Anterior lens capsule, derived from two kinds of capsulorhexis, were randomly divided into two groups according to random number table. The ultrastructure of the capsule edge and its closer tissue were observed by transmission electron microscopy(TEM) and optical microscopy respectively. The surgical completion conditions and corneal endothelial cell counts were analyzed pre-and post-operation after two capsulorhexis.RESULTS: The capsule derived from CCC had smooth edge, well-organized cellular structure and the cells filling into the cutting edge under TEM and optical microscopy. The capsule derived from DHC had an approximate 60 banded area of cell degeneration and necrosis, with dentiform prominences at the edge of the capsule, and no cell structure was observed at this area. The corneal endothelial cell counts of both groups were slightly declined 1 wk post-operation compared with that of preoperation. There was no statistical difference between the two groups(t=1.63, P>0.05).CONCLUSION: DHC shows good clinical value in white mature and hypermature cataract surgery.展开更多
Background:Fuchs endothelial corneal dystrophy is a hereditary disease and the most frequent cause of corneal transplantation in the worldwide.Its main clinical signs are an accelerated decrease in the number of endot...Background:Fuchs endothelial corneal dystrophy is a hereditary disease and the most frequent cause of corneal transplantation in the worldwide.Its main clinical signs are an accelerated decrease in the number of endothelial cells,thickening of Descemet’s membrane and formation of guttae in the extracellular matrix.The cornea’s ability to maintain stromal dehydration is impaired,causing painful epithelial bullae and loss of vision at the point when the amount of corneal endothelial cells cannot be compensated.At present,apart from corneal transplantation,there is no other effective treatment that prevents blindness.Main text:In this review,we first summarized the mutations of COL8A2,TCF4,TCF8,SLC4A11 and AGBL1 genes in Fuchs endothelial corneal dystrophy.The molecular mechanisms associated with Fuchs endothelial corneal dystrophy,such as endoplasmic reticulum stress and unfolded protein response pathway,oxidative stress,mitochondrial dysregulation pathway,apoptosis pathway,mitophagy,epithelial-mesenchymal transition pathway,RNA toxicity and repeat-associated non-ATG translation,and other pathogenesis,were then explored.Finally,we discussed several potential treatments related to the pathogenesis of Fuchs endothelial corneal dystrophy,which may be the focus of future research.Conclusions:The pathogenesis of Fuchs endothelial corneal dystrophy is very complicated.Currently,corneal transplantation is an important method in the treatment of Fuchs endothelial corneal dystrophy.It is necessary to continuously explore the pathogenesis of Fuchs endothelial corneal dystrophy and establish the scientific foundations for the development of next-generation corneal therapeutics.展开更多
Due to the limited capacity of corneal endothelial cells(CECs)division,corneal endothelial diseases have become a great challenge.The cornea is subjected to various mechanical stimuli in vivo,which may have a positive...Due to the limited capacity of corneal endothelial cells(CECs)division,corneal endothelial diseases have become a great challenge.The cornea is subjected to various mechanical stimuli in vivo,which may have a positive or negative influence.Thus,it is significant to gain an insight into the mechanism of mechanobiology of CECs for seeking more possible treatment.The purpose of this study was to determine the impacts of mechanical stretch and substrate stiffness on the morphology and fundamental cell behavior of CECs.Rabbit corneal endothelial cells(RCECs)were subjected to a 5%mechanical stretch or cultured on substrates of different stiffness.The impacts of mechanical stimulus on cell area,aspect ratio,circularity,cell density,nuclear shape,cytoskeleton,and cell viability were investigated.The expressions of the corneal endothelium-related markers ZO-1 and Na^(+)/K^(+) ATPase were also evaluated by confocal immunofluorescence microscopy in the stiffness group.Our results suggested that mechanical stretch promoted the rearrangement of the cytoskeleton while decreasing the cell circularity,nuclear area,and cell density as well as cell viability.RCECs cultured on 10 kPa substrates,which was close to the physiological stiffness of rabbit Descemet's membrane(DM),showed better cell morphology,more stable actin cytoskeleton assembly,and more robust expression of the functional marker compared with other softer or stiffer substrates.In summary,mechanical stretch and substrate stiffness have profound influences on the morphology and function of CECs,which may have implications for the understanding and possible treatment of corneal endothelial diseases.展开更多
文摘Postoperative complications of phacoemulsification,such as corneal edema caused by human corneal endothelial cell(CEC)injury,are still a matter of concern.Although several factors are known to cause CEC damage,the influence of ultrasound on the formation of free radicals during surgery should be considered.Ultrasound in aqueous humor induces cavitation and promotes the formation of hydroxyl radicals or reactive oxygen species(ROS).ROS-induced apoptosis and autophagy in phacoemulsification have been suggested to significantly promote CEC injury.CEC cannot regenerate after injury,and measures must be taken to prevent the loss of CEC after phacoemulsification or other CEC injuries.Antioxidants can reduce the oxidative stress injury of CEC during phacoemulsification.Evidence from rabbit eye studies shows that ascorbic acid infusion during operation or local application of ascorbic acid during phacoemulsification has a protective effect by scavenging free radicals or reducing oxidative stress.Both in experiments and clinical practice,hydrogen dissolved in the irrigating solution can also prevent CEC damage during phacoemulsification surgery.Astaxanthin(AST)can inhibit oxidative damage,thereby protecting different cells from most pathological conditions,such as myocardial cells,luteinized granulosa cells of the ovary,umbilical vascular endothelial cells,and human retina pigment epithelium cell line(ARPE-19).However,existing research has not focused on the application of AST to prevent oxidative stress during phacoemulsification,and the related mechanisms need to be studied.The Rho related helical coil kinase inhibitor Y-27632 can inhibit CEC apoptosis after phacoemulsification.Rigorous experiments are required to confirm whether its effect is realized through improving the ROS clearance ability of CEC.
文摘AIM:To compare the corneal outcome in Fuchs’endothelial dystrophy(FED)patients between femtosecond laser-assisted cataract surgery(FLACS)and conventional phaco surgery(CPS).METHODS:This was a randomized controlled study comparing one eye surgery by FLACS and the contralateral eye operated by CPS(stop and chop technique)in FED patients.Central corneal thickness,corneal light backscatter,corneal densitometry,and central corneal endothelial cell count and hexagonality(noncontact endothelial cell microscope),and corrected distance visual acuity(CDVA)were assessed preoperatively and at day 1,40,and 180 postoperatively.RESULTS:Totally 31 patients(16 women)were included.At day 40 postoperatively,the mean endothelial cell loss(ECL)was 23.67%by FLACS and 17.30%by CPS(P=0.53).At day 180 postoperatively,ECL was 25.58%in FLACS and 21.32%in CPS(P=0.69).Densitometry data in all layers and all annuli from anterior layer to posterior layer in annuli 0-2,2-6,6-10 and 10-12,total densitometry with all layers and all annuli was performed.A significant difference was found in 6-10(posterior layer)at day 1 with-1.42 grayscale units(GSU;95%CI:-2.66 to-0.19,P=0.02).In 10-12(anterior layer,central layer and all layers)at day 40 were significant different with 7.7(95%CI:1.89 to 13.50,P=0.009),3.97(95%CI:0.23 to 7.71,P=0.03),4.73 GSU(95%CI:0.71 to 8.75,P=0.02),respectively.In the remaining parameters we found no difference between the two groups(P>0.05).Three CPS eyes suffered from corneal decompensation.CONCLUSION:There is no significant difference in corneal outcome between FLACS and CPS.Endothelial cell density and pentacam corneal outcome may be inadequate as outcome parameters in FED patients.
基金Supported by National Natural Science Foundation(No.30572011)Natural Science Foundation of Shandong Province(No.ZR2010HQ041)
文摘AIM:To transfect the cat corneal endothelial cells(CECs) with recombinant human β-nerve growth factor gene adeno-associated virus(AAV-β-NGF) and to observe the effect of the expressed β-NGF protein on the proliferation activity of cat CECs.METHODS:The endothelium of cat cornea was torn under the microscope and rapidly cultivated in Dulbecco's modified Eagle's medium(DMEM) to form single layer CECs and the passage 2 endothelial cells were used in this experiment.The recombinant human AAV-β-NGF was constructed.The recombinant human AAV-β-NGF was transferred into cat CECs directly.Three groups were as following:normal CEC control group,CEC-AAV control group and recombinant CECAAV-β-NGF group.Forty-eight hours after transfection,the total RNA was extracted from the CEC by Trizol.The expression of the β-NGF target gene detected by fluorescence quantitative polymerase chain reaction;proliferation activity of the transfected CEC detected at 48 h by MTT assay;the percentage of G1 cells among CECs after transfect was detected by flow cytometry method(FCM);cell morphology was observed under inverted phase contrast microscope.RESULTS:The torn endothelium culture technique rapidly cultivated single layer cat corneal endothelial cells.The self-designed primers for the target gene and reference gene were efficient and special confirmed through electrophoresis analysis and DNA sequencing.Forty-eight hours after transfect,the human β-NGF gene mRNA detected by fluorescence quantitative polymerase chain reaction showed that there was no significant difference between normal CEC control group and CECAAV control group(P>0.05);there was significant difference between two control groups and recombinant CEC-AAV-β-NGF group(P<0.05).MTT assay showed that transfect of recombinant AAV-β-NGF promoted the proliferation activity of cat CEC,while there was no significant difference between normal CEC control group and CEC-AAV control group(P>0.05).FCM result showed that the percentage of G1 cells in CECAAV-NGF group was 76.8% while that in normal CEC control group and CEC-AAV control group was 46.6% and 49.8%.CONCLUSION:Recombinant AAV-β-NGF promotes proliferation in cat CECs by expressing bioactive β-NGF protein in high efficiency and suggests that its modulation can be used to treat vision loss secondary to corneal endotheiiai dysfunction.
文摘Dear Sir,Iam Dr.Jaya Kaushik from the Department of Ophthalmology of the Post Graduate Institute of Medical Education and Research,Chandigarh,India.I write to present a case report of phacoemulsification in a rare case of cataract associated with keratoconus and Fuch’s endothelial
基金Supported by National Natural Science Foundation of China(No.81470608)。
文摘AIM:To investigate the effects of a selective inhibitor of Rho-associated kinase(ROCK),Y-27632,on inbred Wuzhishan porcine corneal endothelial cells(PCECs)in vitro and in vivo studies.METHODS:Primary PCECs were trypsinized from Wuzhishan miniature porcine corneal tissues.The optimal concentration of Y-27632 on PCECs was determined through MTT and 5-ethynyl-2'-deoxyuridine(EdU)-labeling assays.Seven New Zealand rabbits were used as a corneal endothelial dysfunction model,and a PCECs suspension supplemented with Y-27632 was injected into the anterior chamber of the rabbits.The progression of rabbit corneal opacity and edema were observed by slit lamp examination.The rabbits were sacrificed,and rabbit globes were enucleated for trypan blue-alizarin red staining,hematoxylineosin staining,and immunofluorescence analysis.RESULTS:Administration of 100μmol/L Y-27632 facilitated PCECs'proliferation obviously.The rabbit corneas injected with PCECs suspension and 100μmol/L Y-27632 were restored to transparency significantly after 14d.CONCLUSION:The 100μmol/L Y-27632 treatment improves PCECs'proliferation significantly.And our results suggest that Y-27632 and PCECs can be used to treat corneal endothelial dysfunction.
文摘Background:The goal of this project was to analyze the relationship between cell morphology and proteases/proteases inhibitors(PIs)secretion profile in fuchs endothelial corneal dystrophy(FECD)corneal endothelial cells(CECs).Methods:Cell morphology was determined using a circularity index(4π×area/perimeter2)for each CECs population extracted from surgical FECD specimens(N=2)and healthy Eye bank corneas(N=3).CECs were cultured 28 days post-confluency.Supernatant was collected and analysed using Proteome Profiler Array detecting 35 proteases and 32 PIs(R&D Systems).Proteome signal was analyzed using Image Studio Lite and correlated with the population’s circularity index.Results:Calculation of circularity index reported different morphologies among FECD populations(0.59±0.18 and 0.64±0.17)and healthy populations(0.44±0.18,0.66±0.13 and 0.71±0.11).Proteome arrays revealed the presence of 10 proteases(ADAMTS1,Cathepsin A,B,D,and X/Z/P,DPPIV/CD26,MMP-2,3 and 12,uPA/Urokinase)and 10 PIs(Protease Nexin II,Cystatin B and C,EMMPRIN/CD147,Latexin,Lipocalin-1,Serpin E1,TFPI,TFPI-2,TIMP-1,2 and 4).Healthy and FECD specimens showed similar variation patterns according to morphology for secretion of ADAMTS1,MMP-3 and 12.However,opposing patterns between healthy and FECD populations were observed for Cathepsin B and D.Moreover,some proteins did not show variation according to phenotype in healthy CECs,but did in FECD CECs:Cathepsin A,Cystatin C,TFPI-2 and total TIMPs.For the other proteins,secretion did not vary according to morphology or no specific pattern was distinguishable.Conclusions:To conclude,our results suggest that cell phenotype is linked to the secretion of certain proteases/PIs in both groups.However,there seems to be differences in secretion of particular proteases and PIs between FECD and healthy specimens as morphology did not have a similar influence.These differences might initiate an imbalance between proteases and PIs explaining the irregular thickening of the Descemet membrane seen in FECD.
文摘Three months after surgery, the research group showed significantly statistical improvement in visual acuity, a statistically significant decrease in corneal endothelial cell density, a statistically significant increase in the percentage of 5 and 8 sided cells and a statistically significant decrease in the percentage of six sided cells. Central corneal thickness and the percentage of 4 and 7 and more than 8 sided did not change in a statistically significant way. Comparing the test group and control group, no statistically significant differences were detected in the examined parameters. The present study also shows that the cornea in the eyes with congenital cataract does not show statistically significant changes in the density and the morphology of the corneal endothelial cells and the thickness of the cornea and in terms of corneal thickness in comparison to the corneas of healthy eyes. Although in corneas undergoing cataract occurs statistically significant changes, the influence of the cornea does not affect the improvement in visual acuity which was also demonstrated in this study.
基金the National Natural Science Foundation of China for Young Scholars(No.81700872)the National Natural Science Foundation of China(No.81770955)+4 种基金the Project of Shanghai Science and Technology(No.17411950200No.20410710100)the Joint Research Project of New Frontier Technology in Municipal Hospitals(No.SHDC12018103)the Clinical Research Plan of SHDC(No.SHDC2020CR1043B)the Project of Shanghai Xuhui District Science and Technology(No.2020-015)。
文摘AIM:To investigate the safety and efficacy of using a onestep viscoelastic agent technique for posterior chamber phakic implantable collamer lens with a central hole(ICL V4 c)implantation for myopia correction.METHODS:The one-step viscoelastic agent technique for ICL V4 c implantation was used in 100 eyes of 52 patients.Refractive outcomes,intraocular pressure(IOP),corneal endothelial cell,and corneal densitometry values were evaluated at 1 d,1 wk,1 and 3 mo postoperatively.RESULTS:All the surgeries were uneventful.No corrected distance visual acuity was lost after 3 mo.IOP was 16.12±3.18 mm Hg before surgery,and 14.74±3.08 mm Hg at 1d and 14.50±2.56 mm Hg at 3 mo after surgery(P<0.05).Corneal endothelial cell density was 2580±242 cell/mm^(2),the coefficient of variation in cell size was 42.11%±7.92%,and the percentage of hexagonal cells was 40.98%±9.46%before surgery.No significant difference was found when these outcomes were compared between the studied time points(P>0.05).The corneal densitometry values of the central 2 mm and 2 to 6 mm areas showed similar regularities.After surgery,the values significantly increased at 1 d,then decreased to the preoperative values at 1 wk,and then continued to decrease at 3 mo(P<0.05).CONCLUSION:The one-step viscoelastic agent technique for ICL V4c implantation is found to be safe and effective for myopia correction and causes little disturbance to the cornea.
基金Supported by the Shaanxi Provincial Department of Science and Technology Agency Project (No.2017SF-288)
文摘AIM: To compare the corneal endothelial cell counts pre-and post-operation, ultrastructure of anterior lens capsule and surgical completion after diathermic highfrequency capsulorhexis(DHC) and continuous curvilinear capsulorrhexis(CCC) applied in phacoemulsification(PHACO) of white cataract surgery.METHODS: Sixty-six eyes of 66 patients(33 males and 33 females) with cataract aged between 60 and 80 y(mean 72.5±5.5) were recruited and undergone the surgery from June 2014 to November 2016. Anterior lens capsule, derived from two kinds of capsulorhexis, were randomly divided into two groups according to random number table. The ultrastructure of the capsule edge and its closer tissue were observed by transmission electron microscopy(TEM) and optical microscopy respectively. The surgical completion conditions and corneal endothelial cell counts were analyzed pre-and post-operation after two capsulorhexis.RESULTS: The capsule derived from CCC had smooth edge, well-organized cellular structure and the cells filling into the cutting edge under TEM and optical microscopy. The capsule derived from DHC had an approximate 60 banded area of cell degeneration and necrosis, with dentiform prominences at the edge of the capsule, and no cell structure was observed at this area. The corneal endothelial cell counts of both groups were slightly declined 1 wk post-operation compared with that of preoperation. There was no statistical difference between the two groups(t=1.63, P>0.05).CONCLUSION: DHC shows good clinical value in white mature and hypermature cataract surgery.
基金supported by the Heilongjiang Postdoctoral Fund(Grant No.LBH-Z18185)the Heilongjiang Provincial Colleges and Universities Basic Scientific Research Fund(Grant No.2018-KYYWF-0484).
文摘Background:Fuchs endothelial corneal dystrophy is a hereditary disease and the most frequent cause of corneal transplantation in the worldwide.Its main clinical signs are an accelerated decrease in the number of endothelial cells,thickening of Descemet’s membrane and formation of guttae in the extracellular matrix.The cornea’s ability to maintain stromal dehydration is impaired,causing painful epithelial bullae and loss of vision at the point when the amount of corneal endothelial cells cannot be compensated.At present,apart from corneal transplantation,there is no other effective treatment that prevents blindness.Main text:In this review,we first summarized the mutations of COL8A2,TCF4,TCF8,SLC4A11 and AGBL1 genes in Fuchs endothelial corneal dystrophy.The molecular mechanisms associated with Fuchs endothelial corneal dystrophy,such as endoplasmic reticulum stress and unfolded protein response pathway,oxidative stress,mitochondrial dysregulation pathway,apoptosis pathway,mitophagy,epithelial-mesenchymal transition pathway,RNA toxicity and repeat-associated non-ATG translation,and other pathogenesis,were then explored.Finally,we discussed several potential treatments related to the pathogenesis of Fuchs endothelial corneal dystrophy,which may be the focus of future research.Conclusions:The pathogenesis of Fuchs endothelial corneal dystrophy is very complicated.Currently,corneal transplantation is an important method in the treatment of Fuchs endothelial corneal dystrophy.It is necessary to continuously explore the pathogenesis of Fuchs endothelial corneal dystrophy and establish the scientific foundations for the development of next-generation corneal therapeutics.
基金National Natural Science Foundation of China(NSFC)Research Grants(U20A20390,11827803,11402017).
文摘Due to the limited capacity of corneal endothelial cells(CECs)division,corneal endothelial diseases have become a great challenge.The cornea is subjected to various mechanical stimuli in vivo,which may have a positive or negative influence.Thus,it is significant to gain an insight into the mechanism of mechanobiology of CECs for seeking more possible treatment.The purpose of this study was to determine the impacts of mechanical stretch and substrate stiffness on the morphology and fundamental cell behavior of CECs.Rabbit corneal endothelial cells(RCECs)were subjected to a 5%mechanical stretch or cultured on substrates of different stiffness.The impacts of mechanical stimulus on cell area,aspect ratio,circularity,cell density,nuclear shape,cytoskeleton,and cell viability were investigated.The expressions of the corneal endothelium-related markers ZO-1 and Na^(+)/K^(+) ATPase were also evaluated by confocal immunofluorescence microscopy in the stiffness group.Our results suggested that mechanical stretch promoted the rearrangement of the cytoskeleton while decreasing the cell circularity,nuclear area,and cell density as well as cell viability.RCECs cultured on 10 kPa substrates,which was close to the physiological stiffness of rabbit Descemet's membrane(DM),showed better cell morphology,more stable actin cytoskeleton assembly,and more robust expression of the functional marker compared with other softer or stiffer substrates.In summary,mechanical stretch and substrate stiffness have profound influences on the morphology and function of CECs,which may have implications for the understanding and possible treatment of corneal endothelial diseases.
