AIM: To investigate the effects of Msx2 on lens epithelium cell cycle, and evaluate the changes of the proliferation, apoptosis of lens epithelium cells. METHODS: Mice lens epithelium cells were cultured and transfect...AIM: To investigate the effects of Msx2 on lens epithelium cell cycle, and evaluate the changes of the proliferation, apoptosis of lens epithelium cells. METHODS: Mice lens epithelium cells were cultured and transfected with pEGFP-Msx2 and control. Msx2-deficient mice (Msx2(-/-)) lens tissue were isolated. Lens tissue and transfected cells were prepared for mRNA extraction using Trizol reagent. CyclinD1 and Prox1 expression were evaluated by real-time RT-PCR. BrdU incorporation and apoptosis rate were investigated by immunofluorescence and flow cytometry analysis. RESULTS: After transfected with pEGFP-Msx2, lens epithelium cells failed to incorporate BrdU and anti - phospho-histone-3 immunofluorescence failed to detect cell nuclei which GFP were positive. Msx2 over expression resulted in increasing apoptosis rate in lens epithelium cells. CyclinD1 and Prox1 expression increased significantly in Msx2 knockout mice by real time RT-PCR quantization and CyclinD1 expression decreased significantly in Msx2overexpressed cell. CONCLUSION: Msx2 has the effect of inhibiting proliferation and differentiation, triggering apoptosis on mice lens epithelium cells.展开更多
A long life election spin resonance (ESR) signal at g=2.0006 was observed in the normal lens epithelium and cortical fibers. During ultraviolet (UV) exposure, a new ESR signal at g = 2.0060 was found in the lens epith...A long life election spin resonance (ESR) signal at g=2.0006 was observed in the normal lens epithelium and cortical fibers. During ultraviolet (UV) exposure, a new ESR signal at g = 2.0060 was found in the lens epithelium. But this specific signal was not detected in the lens cortical fibers. This suggested that lens epithelial cells were more susceptible to the free radical formation which was induced by UV light. By means of ESR spin probe oxymetry, the oxygen uptake of lens epithelial cells was meas...展开更多
AIM:To explore the DNA methylation of COL4A1 in ultraviolet-B(UVB)-induced age-related cataract(ARC)models in vitro and in vivo.METHODS:Human lens epithelium B3(HLEB3)cells and Sprague Dawley rats were exposure to UVB...AIM:To explore the DNA methylation of COL4A1 in ultraviolet-B(UVB)-induced age-related cataract(ARC)models in vitro and in vivo.METHODS:Human lens epithelium B3(HLEB3)cells and Sprague Dawley rats were exposure to UVB respectively.The MTT assay was utilized to evaluate cell proliferation.Flow cytometry was employed for analysis of cell apoptosis and cell cycle.COL4A1 expression in HLEB3 cells and anterior lens capsules were assessed using Western blot and reverse transcription-polymerase chain reaction(RTPCR).The localization of COL4A1 in HLEB3 cells was determined by immunofluorescence.The methylation status of CpG islands located in COL4A1 promoter was verified using bisulfite-sequencing PCR(BSP).DNMTs and TETs mRNA levels was examined by RT-PCR.RESULTS:UVB exposure decreased HLEB3 cells proliferation,while increased the apoptosis rate and cells were arrested in G0/G1 phase.COL4A1 expression was markedly inhibited in UVB treated cells compared to the controls.Hypermethylation status was detected in the CpG islands within COL4A1 promoter in HLEB3 cells subjected to UVB exposure.Expressions of DNMTs including DNMT1/2/3 were elevated in UVB treated HLEB3 cells compared to that in the controls,while expressions of TETs including TET1/2/3 showed the opposite trend.Results from the UVB treated rat model further confirmed the decreased expression of COL4A1,hypermethylation status of the CpG islands at promoter of COL4A1 and abnormal expression of DNMT1/2/3 and TET1/2/in UVB exposure group.CONCLUSION:DNA hypermethylation of COL4A1 promoter CpG islands is correlated with decreased COL4A1 expression in UVB induced HLEB3 cells and anterior lens capsules of rats.展开更多
AIM： To evaluate human lens epithelium cell apoptosis and epithelial to mesenchymal transition （EMT） induced by femtosecond laser in femtosecond laser assisted cataract surgery （FLACS）. METHODS： Sixty cataract ...AIM： To evaluate human lens epithelium cell apoptosis and epithelial to mesenchymal transition （EMT） induced by femtosecond laser in femtosecond laser assisted cataract surgery （FLACS）. METHODS： Sixty cataract patients with N2 to N3 stage according to the LOCS III were enrolled in this study and divided into three groups randomly： FLACSl group （cataract surgery by FLACS with LenSx）, FLACS2 group （cataract surgery by FLACS with LensAR） and manual group （cataract surgery by phacoemulsification）. Patients in two FLACS groups performed anterior capsulotomy by LenSx or LensAR laser system. Patients in the manual group were performed continuous curvilinear capsulorrhexis （CCC） manually. The anterior capsules were fixed right after moved out of eye. Hematoxylin-eosine staining, immunofluorescence staining and real-time PCR were performed in order to observe human lens epithelium cells changes after cataract surgery. RESULTS： The capsule cutting edge was shown irregularity and roughness in two FLACS groups and smooth edge in manual capsulotomy by pathologic staining. Irregularities of the cell configuration with partly swollen and destroyed nuclei were observed in two FLACS groups. Femtosecond laser could induce a significantly higher cell apoptosis in human lens epithelium cell than manually performed CCC （P〈0.05）. Lens epithelium cells apoptosis were correlated with femtosecond laser duration according to Pearson correlation analysis. Decreased N-cadherin expression, alpha-SMA and FSP-1 level in two FLACS groups showed the inhibition of cell EMT. CONCLUSION： Femtosecond laser may affect the apoptosis and EMT of lens epithelium cells which are under the peeled central lens capsule.展开更多
目的探讨变色人工晶状体（IOL）对可见光诱导人视网膜色素上皮（RPE）细胞损伤的保护作用。方法实验研究。体外培养人RPE细胞（ARPE一19），用同一光照度的可见光照射，在光通路上置不同IOL。RPE细胞分为5组：阴性对照组（无光照，无IO...目的探讨变色人工晶状体（IOL）对可见光诱导人视网膜色素上皮（RPE）细胞损伤的保护作用。方法实验研究。体外培养人RPE细胞（ARPE一19），用同一光照度的可见光照射，在光通路上置不同IOL。RPE细胞分为5组：阴性对照组（无光照，无IOL）、阳性对照组（照光，无IOL）、无色IOL组（照光，FV-60AIOL）、变色IOL组（照光，BV-60AIOL）、黄色IOL组（照光，Acrys of Natural IOL）。用MTT法检测RPE细胞活力，透射电子显微镜、脱氧核糖核苷酸末端转移酶介导的缺口末端标记（TUNEL）检测RPE细胞凋亡情况；Hoechst染色计算凋亡指数（AI）；酶联免疫吸附法检测血管内皮生长因子（VEGF）、色素上皮衍生因子（PEDF）的浓度。对数据进行方差分析，多个均数间两两比较用q检验。结果阴性对照组、阳性对照组、无色IOL组、变色IOL组、黄色IOL组细胞吸光度（A）值分别为0．61±0．01、0．39±0．01、0．46±0．01、0．57±0．01及0．58±0．02；TUNEL染色和透射电镜下检查均发现光照后细胞出现凋亡改变。5个组细胞的AI值分别是（0．99±0．65）％、（9．28±0．59）％、（6．54±0．37）％、（4．69±0．74）％及（2．72±0．67）％。5个组VEGF浓度分别为（3931．64±25．26）ng／L、（5459．38±42．69）ng／L、（4800．55±42．12）ng／L、（4561．21±46．27）ng／L及（4420．35±28．66）ng/L；PEDF浓度分别为（182．85±1．68）μg／L、（156．52±0．64）μg／L、（155．57±3．43）μg／L、（167．57±2．88）μg／L及（170．90±2．99）μg／L。变色IOL组与黄色IOL组比较，细胞活力（q=0．94）、PEDF浓度（q=1．51）的差异无统计学意义（P〉0．05）；AI值（q=5．04）、VEGF浓度（q=6．78）的差异有统计学意义（P〈0．05）；变色IOL组与无色IOL组比较，细胞活力（q=18．92）、AI值（q=4．78）、VEGF浓度（q=11．52）及PEDF浓度（q=4．96）的差异有统计学意义（P〈0．05）。结论变色IOL能减轻RPE细胞的急性光损伤程度，保护作用优于紫外线阻断型IOL，弱于蓝光滤过型IOL。展开更多
基金Science Foundation of Liaoning Province,China (No. 2011225014)
文摘AIM: To investigate the effects of Msx2 on lens epithelium cell cycle, and evaluate the changes of the proliferation, apoptosis of lens epithelium cells. METHODS: Mice lens epithelium cells were cultured and transfected with pEGFP-Msx2 and control. Msx2-deficient mice (Msx2(-/-)) lens tissue were isolated. Lens tissue and transfected cells were prepared for mRNA extraction using Trizol reagent. CyclinD1 and Prox1 expression were evaluated by real-time RT-PCR. BrdU incorporation and apoptosis rate were investigated by immunofluorescence and flow cytometry analysis. RESULTS: After transfected with pEGFP-Msx2, lens epithelium cells failed to incorporate BrdU and anti - phospho-histone-3 immunofluorescence failed to detect cell nuclei which GFP were positive. Msx2 over expression resulted in increasing apoptosis rate in lens epithelium cells. CyclinD1 and Prox1 expression increased significantly in Msx2 knockout mice by real time RT-PCR quantization and CyclinD1 expression decreased significantly in Msx2overexpressed cell. CONCLUSION: Msx2 has the effect of inhibiting proliferation and differentiation, triggering apoptosis on mice lens epithelium cells.
基金The work was sponsored by National Natural Science Foundation of China (NSFC)
文摘A long life election spin resonance (ESR) signal at g=2.0006 was observed in the normal lens epithelium and cortical fibers. During ultraviolet (UV) exposure, a new ESR signal at g = 2.0060 was found in the lens epithelium. But this specific signal was not detected in the lens cortical fibers. This suggested that lens epithelial cells were more susceptible to the free radical formation which was induced by UV light. By means of ESR spin probe oxymetry, the oxygen uptake of lens epithelial cells was meas...
基金Supported by the Shaanxi Provincial Department of Science and Technology Agency Project(No.2022SF-502)Xi’an Medical University Scientific Research Capacity Improvement Project(No.2022NLTS104)+2 种基金The Fifth Batch of Key Disciplines of Xi’an Medical University(No.medical technology12202306)Yunzhen Optometry Special Fund(No.2021HXZR10)Innovation and Entrepreneurship Training Program for College Students(No.S202211840043).
文摘AIM:To explore the DNA methylation of COL4A1 in ultraviolet-B(UVB)-induced age-related cataract(ARC)models in vitro and in vivo.METHODS:Human lens epithelium B3(HLEB3)cells and Sprague Dawley rats were exposure to UVB respectively.The MTT assay was utilized to evaluate cell proliferation.Flow cytometry was employed for analysis of cell apoptosis and cell cycle.COL4A1 expression in HLEB3 cells and anterior lens capsules were assessed using Western blot and reverse transcription-polymerase chain reaction(RTPCR).The localization of COL4A1 in HLEB3 cells was determined by immunofluorescence.The methylation status of CpG islands located in COL4A1 promoter was verified using bisulfite-sequencing PCR(BSP).DNMTs and TETs mRNA levels was examined by RT-PCR.RESULTS:UVB exposure decreased HLEB3 cells proliferation,while increased the apoptosis rate and cells were arrested in G0/G1 phase.COL4A1 expression was markedly inhibited in UVB treated cells compared to the controls.Hypermethylation status was detected in the CpG islands within COL4A1 promoter in HLEB3 cells subjected to UVB exposure.Expressions of DNMTs including DNMT1/2/3 were elevated in UVB treated HLEB3 cells compared to that in the controls,while expressions of TETs including TET1/2/3 showed the opposite trend.Results from the UVB treated rat model further confirmed the decreased expression of COL4A1,hypermethylation status of the CpG islands at promoter of COL4A1 and abnormal expression of DNMT1/2/3 and TET1/2/in UVB exposure group.CONCLUSION:DNA hypermethylation of COL4A1 promoter CpG islands is correlated with decreased COL4A1 expression in UVB induced HLEB3 cells and anterior lens capsules of rats.
文摘AIM： To evaluate human lens epithelium cell apoptosis and epithelial to mesenchymal transition （EMT） induced by femtosecond laser in femtosecond laser assisted cataract surgery （FLACS）. METHODS： Sixty cataract patients with N2 to N3 stage according to the LOCS III were enrolled in this study and divided into three groups randomly： FLACSl group （cataract surgery by FLACS with LenSx）, FLACS2 group （cataract surgery by FLACS with LensAR） and manual group （cataract surgery by phacoemulsification）. Patients in two FLACS groups performed anterior capsulotomy by LenSx or LensAR laser system. Patients in the manual group were performed continuous curvilinear capsulorrhexis （CCC） manually. The anterior capsules were fixed right after moved out of eye. Hematoxylin-eosine staining, immunofluorescence staining and real-time PCR were performed in order to observe human lens epithelium cells changes after cataract surgery. RESULTS： The capsule cutting edge was shown irregularity and roughness in two FLACS groups and smooth edge in manual capsulotomy by pathologic staining. Irregularities of the cell configuration with partly swollen and destroyed nuclei were observed in two FLACS groups. Femtosecond laser could induce a significantly higher cell apoptosis in human lens epithelium cell than manually performed CCC （P〈0.05）. Lens epithelium cells apoptosis were correlated with femtosecond laser duration according to Pearson correlation analysis. Decreased N-cadherin expression, alpha-SMA and FSP-1 level in two FLACS groups showed the inhibition of cell EMT. CONCLUSION： Femtosecond laser may affect the apoptosis and EMT of lens epithelium cells which are under the peeled central lens capsule.
文摘目的探讨变色人工晶状体（IOL）对可见光诱导人视网膜色素上皮（RPE）细胞损伤的保护作用。方法实验研究。体外培养人RPE细胞（ARPE一19），用同一光照度的可见光照射，在光通路上置不同IOL。RPE细胞分为5组：阴性对照组（无光照，无IOL）、阳性对照组（照光，无IOL）、无色IOL组（照光，FV-60AIOL）、变色IOL组（照光，BV-60AIOL）、黄色IOL组（照光，Acrys of Natural IOL）。用MTT法检测RPE细胞活力，透射电子显微镜、脱氧核糖核苷酸末端转移酶介导的缺口末端标记（TUNEL）检测RPE细胞凋亡情况；Hoechst染色计算凋亡指数（AI）；酶联免疫吸附法检测血管内皮生长因子（VEGF）、色素上皮衍生因子（PEDF）的浓度。对数据进行方差分析，多个均数间两两比较用q检验。结果阴性对照组、阳性对照组、无色IOL组、变色IOL组、黄色IOL组细胞吸光度（A）值分别为0．61±0．01、0．39±0．01、0．46±0．01、0．57±0．01及0．58±0．02；TUNEL染色和透射电镜下检查均发现光照后细胞出现凋亡改变。5个组细胞的AI值分别是（0．99±0．65）％、（9．28±0．59）％、（6．54±0．37）％、（4．69±0．74）％及（2．72±0．67）％。5个组VEGF浓度分别为（3931．64±25．26）ng／L、（5459．38±42．69）ng／L、（4800．55±42．12）ng／L、（4561．21±46．27）ng／L及（4420．35±28．66）ng/L；PEDF浓度分别为（182．85±1．68）μg／L、（156．52±0．64）μg／L、（155．57±3．43）μg／L、（167．57±2．88）μg／L及（170．90±2．99）μg／L。变色IOL组与黄色IOL组比较，细胞活力（q=0．94）、PEDF浓度（q=1．51）的差异无统计学意义（P〉0．05）；AI值（q=5．04）、VEGF浓度（q=6．78）的差异有统计学意义（P〈0．05）；变色IOL组与无色IOL组比较，细胞活力（q=18．92）、AI值（q=4．78）、VEGF浓度（q=11．52）及PEDF浓度（q=4．96）的差异有统计学意义（P〈0．05）。结论变色IOL能减轻RPE细胞的急性光损伤程度，保护作用优于紫外线阻断型IOL，弱于蓝光滤过型IOL。
文摘目的探讨空中乘务员配戴角膜接触镜（contact lens，CL）对其角膜上皮损伤的影响及可能机制。方法回顾分析2015年1月至2016年10月在中国南方航空有限公司航卫中心行体检鉴定的配戴角膜接触镜的707例空中乘务员体检资料。根据受检者角膜接触镜史，将其分为3组：短期配戴组（配戴时间〈1年）、中期配戴组（配戴时间1～3年）、长期配戴组（配戴时间3～5年）。受检者均行裂隙灯显微镜、泪膜破裂时间（tear film break-up time，TBUT）、角膜荧光素染色、角膜知觉等检查。根据角膜上皮损伤的诊断标准及病变程度分级，统计空中乘务员角膜上皮损伤的检出率及其眼部体征（结膜充血、角膜上皮病变、泪膜异常、角膜知觉减退）的分布情况，并比较3个不同戴镜时间组间的上述差异。结果707例空中乘务员中结膜充血479例（67．75％）、角膜上皮病变116例（16．41％）、泪膜异常20例（2．83％）、角膜知觉减退12例（1．70％）。诊断为角膜上皮损伤116例，其中轻度88例、中度17例、重度儿例。3组间结膜充血、角膜上皮病变检出率的差异均有统计学意义（χ^2=94．094、10．035，P〈0．01）。3个不同戴镜时间组角膜上皮损伤的检出率分别为10．96％、15．26％、21．84％，且组间差异有统计学意义（χ^2=10．035，P〈0．05）；中重度角膜上皮损伤疾病构成比分别为13．04％、22．22％、29．82％。结论空中乘务员配戴角膜接触镜可以引起角膜上皮损伤，且随着配戴时间的延长，角膜上皮损伤的检出率明显增高、病变程度有加重趋势。角膜上皮损伤可能与角膜接触镜的机械损伤、炎症反应、泪膜异常等因素有关。
