ABSTRACT
AIM: To investigate the role and mechanism of N6-methyladenosine(m6A)methyltransferase 3(METTL3)in the pathogenesis of diabetic cataract.METHODS: We cultured SRA01/04 cells in low and high sugar media for 24h and measured changes in epithelial-mesenchymal transition(EMT)indicators(E-Cadherin, N-Cadherin, ZO-1 and α-SMA)using RT-qPCR and Western blot assays. Cell migration was also assessed using transwell and scratch assays. To investigate the expression level and localization of METTL3 in human lens anterior capsules tissues. Additionally, we used m6A dot blot assay to detect the m6A methylation level of cells cultured in low and high glucose media for 24h, and employed RT-qPCR and Western blot experiments to detect RNA and protein expression of METTL3 in cells. We then treated the cells with METTL3 inhibitor and measured changes in EMT markers by RT-qPCR and Western blot; m6A methylation level was detected by m6A dot blot test; cell migration was detected by Transwell. Finally, the expression of transforming growth factor-β(TGFβ1)in cultured cells was assessed by immunofluorescence staining and the expression levels of TGFβ1 and SNAIL in cells were determined using RT-qPCR and Western blot.RESULTS: Under high glucose conditions, the expression of EMT markers, METTL3, and m6A methylation levels were significantly increased in cells(P<0.05). Furthermore, the migratory ability of cells was higher in high-sugar medium than in low-sugar medium. In human lens anterior capsules, METTL3 expression was higher in patients with diabetic cataract compared to those with age-related cataract. Importantly, treatment with the METTL3 inhibitor STM2457 inhibited EMT in cells, the expression of TGFβ1 and SNAIL, as well as m6A methylation levels in cells(all P<0.05)compared to high-sugar + dimethyl sulfoxide(DMSO)group. Moreover, the migratory capacity of cells was reduced after the addition of STM2457 compared to the high-sugar + DMSO group.CONCLUSION:METTL3 promotes the EMT in human lens epithelial cells under high glucose conditions by activating the TGFβ1/SNAIL pathway, thus contributing to the development of diabetic cataracts.
ABSTRACT
AIM: To investigate the role and mechanism of methyltransferase-like 3(METTL3)-mediated N6-methyladenosine(m6A)methylation modification in regulating biological activity of vascular endothelial cells in the pathogenesis of choroidal neovascularization.METHODS: Human umbilical vein endothelial cells(HUVEC)cultured in vitro were divided into the following groups: control group(normal culture), low density lipoprotein(LDL)group, fluorescence-labelled LDL(Dil-LDL)group, 12.5μg/mL and 25μg/mL oxidized LDL(ox-LDL)groups, 12.5μg/mL and 25μg/mL fluorescence-labelled ox-LDL(Dil-ox-LDL)groups, DMSO group, STM2457(METTL3 inhibitor)group, DAPT group; and monkey retina-choroidal endothelial cells(RF/6A)cultured in vitro were divided into control group, DMSO group, 12.5 μg/mL ox-LDL group, and DAPT group. Endocytosed lipoprotein level was examined through fluorescence microscopy. RNA m6A methylation level was detected through a dot blot assay. Protein and RNA levels of METTL3 or angiogenesis-related markers were measured through Western blot assays and real-time quantitative polymerase chain reaction(RT-qPCR), respectively. METTL3 expression and localization were investigated through immunofluorescence. Cell migratory and tube formation capacities were assessed through transwell migration and tube formation assays, respectively.RESULTS: Endocytosed lipoprotein levels in HUVECs exposed to Dil-LDL, 12.5μg/mL and 25μg/mL Dil-ox-LDL groups were significantly higher than those in the control group. 12.5μg/mL and 25μg/mL ox-LDL groups significantly increased m6A methylation(all P<0.05), METTL3 protein expression(all P<0.01), and cell migration and angiogenesis capacities(all P<0.01). METTL3 mRNA level was significantly unregulated in the 12.5μg/mL ox-LDL group(P<0.05). In comparison to the DMSO group, the addition of STM2457 caused significant decrease in m6A methylation level(P<0.05), expression of VEGF and other angiogenesis-related markers(all P<0.05), cell migration and angiogenesis capacities(all P<0.01)and the expression of NICD(P<0.05). However, there were no significant differences in METTL3 protein and mRNA levels(all P>0.05). The expression of VEGF and NICD(all P<0.05), as well as the ability of cell migration and angiogenesis of RF/6A, was all significantly decreased in the DAPT group compared to the DMSO group(all P<0.01).CONCLUSION: METTL3-mediated m6A methylation modification promotes angiogenesis in vascular endothelial cells via the Notch signaling pathway in the pathogenesis of choroidal neovascularization.
ABSTRACT
Objective Ultraviolet radiation can induce the expression of matrix metalloproteinases (MMPs) in pterygium epithelial cells. To investigate the effects of curcumin on the expression of MMP 2 and MMP 9 in human pterygium fibroblasts (HPFs) on UVA-irradiation and its possible mechanism.Methods After optimizing the dose of UVA irradiation and the concentration of curcumin, HPFs in the second generation were divided into control group (no exposure to UVA, no medication), UVA group (exposure to UVA), and UVA + curcumin group (exposure to UVA + curcumin). MMP-2 and MMP-9 levels were tested by zymography. The mRNA expression of MMP-2 and MMP-9 was detected by RT-PCR. The NF-κB-DNA binding activity was detected by electrophoretic mobility shift assay (EMSA).Results Compared with the control group, the secretion of MMP-2 and MMP-9 in the UVA group was increased significantly (P<0.05), which could be suppressed by curcumin (P<0.05). The MMP-9 mRNA levels were also significantly increased in UVA group \[(100±0)% vs (247.0±10.8)%, P<0.05\], and could be inhibited by curcumin \[(88.7±5.1)% vs (247.0±10.8)%, P<0.05\]. The NF-κB-DNA binding was remarkably increased in UVA group (P<0.05), and significantly decreased after treatment with curcumin (P<0.05).Conclusion Curcumin can inhibit the expression of MMP-2 and MMP-9 in UVA-irradiated HPFs, and the mechanism involved in this protective effect might be its inhibitory effect on NF-κB activation.
ABSTRACT
Objective Systemic lupus erythematosus (SLE) cause irreversible damage to multiple organs of the body.This article evaluates the changes in tear film quality and visual quality in patients with SLE.Methods In the study,we selected 66 patients with SLE (66 right eyes) for the study group and 59 healthy volunteers (59 right eyes) for the control group,The measurements of BUT,corneal fluorescein staining(FL) score and SIt were taken in turn for both two groups.Anterior segment OCT was used for measuring the CET,CCT,TMD,TMH and TMA.OQAS Ⅱ system was taken for MTFcutoff,SR,static OSI,OV (100％,20％,9％) and dynamic OSI.Differences of the parameters were compared between the two groups.The data correlation was assessed by Spearman rank correlation analysis.Results There were significant differences in SIt,BUT,FL score,CET,TMH,TMD and TMA between the two groups(P＜0.05),but no significant difference in CCT (P＞0.05).The differences of static OSI,MTFcutoff,SR,OV100％,OV20％,OV9％ between the two groups were statistically significant(P＜0.01).The △OSI(differences between dynamic and static OSI) had the difference between the two groups(t=4.615,P＜0.001).TMA was positively correlated with CET,SIt,and BUT (P＜0.05),and had no correlation with OSI,MTFcutoff,SR,OV100％,OV20％,OV9％ (P＞0.05).Anti-ds-DNA is positively correlated with SIt,TMA,MTFcutoff and SR (r =0.346,0.208,0.257,0.197,P＜0.05) and negatively correlated with OSI(r=-0.292,P＜0.05).There was no correlation between ANA,C3,C4 and the parameters of tear film and visual quality (P＞0.05).Conclusion SLE may affect the amount and stability of the patient's tears and affects the corneal epithelial thickness and visual quality.
ABSTRACT
Orthokeratology is a kind of rigid contact lens which have reverse geometric desi, with higher oxygen permeability and security.Overnight wearing of orthokeratology can decrease the central corneal curvature and increase peripheral corneal curvature by flatting the central department of corneal, thus reduce the refraction of myopia.Through a period of time of wearing orthokeratology lens, patients can obtain good eyesight without frame glasses.Insisting on wearing orthokeratology lens can control the development of myopia.Orthokeratology is widely applied in the control of juvenile myopia, so we need scientific evaluation system to measure the visual quality after wearing orthokeratology lens.Here are the methods that used to evaluate the visual quality after wearing orthokeratology lens.
ABSTRACT
AIM:To evaluate the effect of rigid gas permeable ( RGP ) lens on the tear film and the visual quality in adolescents with a double-pass system. METHODS:This was a prospective study comprised 23 myopia patients ( 39 eyes ) wearing rigid gas permeable contact lens ( RGP ) between Jun. 2015 and Aug. 2015 in the Department of Ophthalmology of Jinling Hospital. Uncorrected visual acuity ( UCVA ) , sphere refractive, cylinder refractive, Schirmer I test, tear film break-up time ( BUT ) , the visual quality and the tear film quality with OQAS II were obtained before and 1wk, 1, 3, 6mo after wearing. The data was analyzed using t-test and One-way. RESULTS:The UCVA in before and after 1wk, 1, 3, 6mo were 0. 23±0. 10, 0. 81±0. 23, 0. 99±0. 11, 1. 01±0. 09, 0. 95±0. 14, the difference was statistically significant ( F=723. 36, P0. 05). Schirmer I test showed no statistically significant difference before and after wearing RGP ( F=4.88, P>0. 05). The stability of tear film was reduced after wearing RGP, and the difference was statistically significant (F=135. 11, P<0. 01). The modulate transfer function ( MTF ) cut- off frequency was reduced after wearing RGP, the object scatter index ( OSI ) was increased after wearing RGP, and the difference was statistically significant, the tear film quality OSI was increased after wearing RGP, and the difference was statistically significant (F=6. 16, 19. 23, 10. 62, P<0. 01).CONCLUSION:The UCVA is significantly increased after wearing RGP. However, there was no significant change in the base-line of tear secretion, but BUT is significantly reduced after wearing RGP. The stability of the tear film is reduced, which can lead to a decline in visual quality.
ABSTRACT
AIM: To evaluate the efficacy of phacoemulsification and intraocular lens implantation surgical intervention of complicated cataract in patients with uveitis. ●METHODS:Retrospective study. A total of 57 cases (57 eyes ) with complicated cataract with uveitis were involved in the study from Jan. 2015 to Dec. 2015. All cases underwent phacoemulsification and intraocular lens implantation surgery successfully. The postoperative reaction, effect, complications and so on were retrospectively analyzed after phacoemulsification and intraocular lens implantation surgery. The date of visual outcome was analyzed using Non - parametric Wilcoxon test. ●RESULTS: lris were bleed in 21 eyes ( 37%) , 4 eyes ( 7%) with posterior capsule rapture and posterior chamber intraocular lens was not implanted in 4 eyes ( 7%) . The uncorrected visual acuity ( UCVA ) was significantly increased after surgery. The UCVA of 8 eyes (14%) were 0. 1 or better before surgery, and the UCVA of 42 eyes (74%) were 0. 1 or better 3mo after surgery, the difference was statistically significant ( Z=23. 42, P ● CONCLUSION: The phacoemulsification combined intraocular lens implantation surgical intervention of complicated cataract in patients with uveitis has good effect and fewer complications.
ABSTRACT
?AIM: To evaluate the effects on recovering of corneal wound and postoperative discomfort of different methods for primary pterygium.?METHODS: Forty-seven cases ( 60 eyes ) of primary pterygium were excised under microscope with limbal epithelial transplantation, with sharp dissection ( 24 cases, 30 eyes, Group A) and blunt dissection (23 cases, 30 eyes, Group B).All cases were followed up for 1d to 1mo.?RESULTS: The recovering of corneal wound was better in Group B on 1st day and 3rd day after surgery.Pain, photophobia and tears, foreign body sensation were more serious in group A on 1st day after surgery with a statistically significant difference (P=0.005,0.015,0.012). Pain, photophobia and tears, foreign body sensation were more serious in Group A on 3rd day after surgery with a statistically significant difference ( P=0.019,0.018, 0.015).There was no statistically significant difference on 1wk and 1mo after surgery (P>0.05).? CONCLUSION: Compared with sharp dissection, primary pterygium excised with blunt dissection can significantly improve recovering of corneal wound and postoperative discomfort.
ABSTRACT
To evaluate the influence of phacoemulsification on corneal endothelium of senile cataract with lupus nephritis (LN). ●METHODS:This clinical trial involved 40 cataract patients with lupus nephritis (40 eyes), and 50 cases (50 eyes) without lupus nephritis. All of them underwent phacoemulsification+lOL implantation. The parameters of corneal endothelial cell including central corneal endothelium cell density ( CED ), average area of endothelial cell ( AVE) and coefficient of variation ( CV) were recorded by corneal endothelial microscope pre -operation and at one month after operation. The data were analyzed by SPSS 13. 0 statistical software. ● RESULTS: Both LN group and control group, the morphology of coneal endothelial was statistical significant differences between pre - operation and 1mo postoperation. The CED was lower, AVE and CV were higher ( P ● CONCLUSlON: The corneal endothelial cell in lupus nephritis patients is more fragile. Safe and reliable operation should be selected for these patients.
ABSTRACT
AIM:To discuss Daidzein intravitreal injection whether has protective and recovery effects on acute nerve damages. METHODS:After the crush models of acute optic nerve were set up, 72 males SD rats were divided into 4 groups randomly as common group without surgery, FBS negative control group, Daidzein treatment group ( 10μmol/L, 100μmol/L, 1000μmol/L ) and positive control group using rats nerve growth factor ( mNGF, 100ng/mL ). Three days after interference, all experimental animals were executed. HE staining was used to evaluate morphologic change of the retina, immunohisochemical staining and western-blot tests for identifying and quantifying the distinct expression of Caspase-3 and GAP-43 among the groups. RESULTS: Compared with the normal group and negative control group, retinal morphology of different concentrations of each Daidzein treatment group and positive control group was more complete, the expression of Caspase-3 protein was relatively lower, the expression of GAP-43 protein was relatively higher, the differences have statistically significance (P<0. 05).CONCLUSION: Daizein injection in the vitreous cavity has the capacity of protection and restoration in rat's acute nerve damages.
ABSTRACT
Background Phlyctenular ophthalmia is a delayed hypersensitivity reaction to some microprotein and affected mainly by adolescent in high incidence.Objective This study was to investigate the microscopy findings of phlyctenular ophthalmia and evaluate the histological changes by laser scanning confocal microscope.Methods Twenty-nine eyes suffered from phlyctenular ophthalmia and twenty normal eyes were examined using laser scanning confocal microscope.The pictures were taken by a CCD camera.All the cases had initially chest X-ray,tuberculin test,bacterial and mycobacteria culture.Results Dendritic and inflammatory cells were increased and concentrated in conjunctiva,and epithelial cells were deformed and squamatizated.The capillaries engorged and the goblet cells were injured.The corneoscleral Vogt meshing of the phlyctenular keratitis was obscured and dendritic cells were intruded into the corneas.The corneal epithelium of phlyctenular keratitis was absent and the subepithelial nerve plexus were bended and fractured,and the dendritic and inflammatory cells were intruded.Scarring of corneal stroma was seen under the laser scanning confocal microscope.Conclusions Laser scanning confocal microscopy is valuable for basic research and clinical diagnosis of phlyctenular ophthalmia.
ABSTRACT
BackgroundPterygium is an ocular surface disease of abnormal cell proliferative kind and angiogenesis plays an important role in its development and recurrence.Several anti-angiogenic therapies have been used to treat pterygium,but there very few studtes for the in vivo observation of the microvessles in pterygium.ObjectiveThis study was to observe angiogenesis in pterygium with a high-resolution confocal microscope in vivo and to perform immunohistochemical study in vitro.MethodsA prospective case-controlled study was designed.Twenty eyes of 20 consecutive patients with primary pterygia and 20 age- and sex-matched patients with inner eye diseases and strabismus with normal conjunctiva were enrolled in this study.An in vivo confocal microscopy imaging system (Heidelberg Retina Tomograph Ⅱ Rostock Cornea Module) was used to collect microvascular pictures from the anterior part of pterygia and normal nasal conjunctiva of controls,and then immunochemistry was performed to examine the expression of CD31 in microvessel in vitro.The vascular density values were compared between these two groups.The correlation of vascular density values between in vivo Heidelberg Retina Tomograph and in vitro immunohistochemistry was calculated.Written informed consent was obtained from pationts before any examination and surgery.ResultsUnder the in vivo confocal microscope,the microvessel density was (8929±2993) μm/mm2 and (4202 ±692)μm/mm2,respectively,in pterygium and the normal conjunctiva group with a statistically significant difference between them (t =6.881,P＜0.01 ).Immunochemistry revealed that the expression of CD31 to measure vascular density was ( 21.00 ± 4.06/400 × field ) and ( 6.07 ± 1.75/400 × field ) in pterygium and the normal conjunctiva group,showing significant difference (t =12.312,P＜0.01 ).Positive correlations were found in the vascular density values between in vivo corneal laser confocal microscopy examination and in vitro immunochemistry examination in both the pterygium group and normal conjunctiva group (pterygium group:r=0.649,P＜0.01 ;normal conjunctiva group: r=0.572,P＜0.01 ) ConclusionsIn vivo confocal microscopy imaging is superior to in vitro immunochemistry in evaluating the microvessel of pterygium.The results of this study offer a new way index for further investigation of the biological behavior of pterygium and its mechanism.
ABSTRACT
AIM:To compare the contrast sensitivity obtained through anterior surface modified prolate intraocular lens with the contrast sensitivity obtained through standard spherical intraocular Iens.METHODS:Patients presenting for cataract surgery in one eye were randomized to receive either the Acrysef IQ intraocular lens(AIcon)or the AR40e intraocular lens(AMO).50 eyes after phacoemulsification with IOL implantation were included,23 eyes received Acrysof IQ,and 27 eyes received AP40e intraocular lens (AMO).Contrast sensitivity testing under photopic conditions (85cd/m2)served as the prindpal outcome measure.The contrast sensitivity function was measured with CSV-1000E Vision Testing Instrument(provided by VECTOR ⅥSION COMPANY) at 30 days after surgery.RESULTS:The Acrysof IQ intraocular lens provided statistically significantly better contrast sensitivity at 6,12 and 18 cydes per degree under photopic conditions.CONCLUSION:The use of anterior surface modified prolate intraocular lens for cataract surgery has the potential to improve functional vision.
ABSTRACT
· AIM: To inspect and compare the functional vision of an aspheric intraocular lens (Tecnis) with those of conventional monofocal silicone and acrylic intraocular lens and multifocal intraocular lens (Array).· METHODS: The IOLs were tested in the eye model, which was designed to be optically equivalent to the theoretical eye model. The eye model is a combination of a spherical photographic lens with 35mm focal length ( IOL put in a water cell)and a charge coupled device (CCD) camera. The images constructed by the lenses are observed on a monitor of personal computer and the contrasts of the images are analyzed by using commercial image processing software.· RESULTS: The modulation transfer function of the eye model equals the scale produced by the theoretical eye model. The images constructed by changing the diameter of aperture stop and IOL.· CONCLUSION: The proposed eye model is useful for testing functional vision and for inspecting differences of intraocular lens.