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1.
Am J Ophthalmol ; 2021 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-34818515

RESUMO

PURPOSE: To evaluate the diagnostic yield of congenital ectopia lentis (EL) in a Chinese cohort by combining panel-based next-generation sequencing with clinical findings DESIGN: A cohort study. METHODS: In total, 175 patients with congenital EL and their available family members (n = 338) were enrolled. All congenital EL patients underwent genetic testing. Genotype-phenotype analyses were conducted to assess the biometric and structural ocular manifestations of congenital EL. RESULTS: In total, 175 patients with congenital EL and 338 of their relatives were included in this study. In these patients, 92.57% (162/175) of disease-related variants were detected in FBN1 (83.43%), CPAMD8 (1.71%), COL4A5 (0.57%), ADAMTSL4 (3.43%), LTBP2 (1.71%), and CBS (2.29%). Based on genetic and clinical findings, the primary diagnostic rate was increased to 40.57% from 19.43% with the exception of the 91 diagnoses of potential MFS, with a new diagnostic strategy for congenital ectopia lentis thus having been developed. Within this group of patients harboring FBN1 mutations, 16.44% (19/141) probands were diagnosed with ectopia lentis syndrome (ELS) and 2.13% (3/141) were diagnosed with Marfan syndrome (MFS). CONCLUSIONS: The results of this cohort study expand the genomic landscape associated with congenital EL in Chinese cohorts. FBN1 mutations represent the most common cause of congenital EL in this population, and we have developed a new diagnostic strategy for congenital EL subtypes via the use of a well-designed panel-based next-Generation Sequencing (NGS) which can be used to efficiently and precisely diagnose patients with congenital EL in a cost-effective manner.

2.
Hum Mutat ; 42(12): 1637-1647, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34550612

RESUMO

Mutations of fibrillin-1 (FBN1) have been associated with Marfan syndrome and pleiotropic connective tissue disorders, collectively termed as "type I fibrillinopathy". However, few genotype-phenotype correlations are known in the ocular system. Patients with congenital ectopia lentis (EL) received panel-based next-generation sequencing, complemented with multiplex ligation-dependent probe amplification. In a total of 125 probands, the ocular phenotypes were compared for different types of FBN1 mutations. Premature termination codons were associated with less severe EL and a thinner central corneal thickness (CCT) than the inframe mutations. The eyes of patients with mutations in the C-terminal region had a higher incidence of posterior staphyloma than those in the middle and N-terminal regions. Mutations in the TGF-ß-regulating sequence had larger horizontal corneal diameters (white-to-white [WTW]), higher incidence of posterior staphyloma, but less severe EL than those with mutations in other regions. Mutations in the neonatal region were associated with thinner CCT. Longer axial length (AL) was associated with mutations in the C-terminal region or TGF-ß regulating sequence after adjusting for age, EL severity, and corneal curvature radius. FBN1 genotype-phenotype correlations were established for some ocular features, including EL severity, AL, WTW, CCT, and so forth, providing novel perspectives and directions for further mechanistic studies.

3.
Int J Ophthalmol ; 14(8): 1218-1224, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34414087

RESUMO

AIM: To investigate whether the axial length (AL)/total corneal refractive power (TCRP) ratio is a sensitive and simple factor that can be used for the early diagnosis of Marfan's syndrome (MFS) in children. METHODS: The relationship between the AL/TCRP ratio and the diagnosis of MFS for 192 eyes in 97 children were evaluate. The biological characteristics, including age, sex, AL, and TCRP, were collected from medical records. Receiver operating characteristic (ROC) curve analysis was performed to investigate whether the AL/TCRP ratio effectively distinguishes MFS from other subjects. The Youden index was used to re-divide the whole population into two groups according to an AL/TCRP ratio of 0.59. RESULTS: Of 96 subjects (mean age 7.46±3.28y) evaluated, 56 (110 eyes) had a definite diagnosis of MFS in childhood based on the revised Ghent criteria, 41 (82 eyes) with diagnosis of congenital ectopia lentis (EL) were included as a control group. AL was negatively correlated with TCRP, with a linear regression coefficient of -0.36 (R 2=0.08). A significant correlation was found between age and the AL/TCRP ratio (P=0.023). ROC curve analysis showed that the AL/TCRP ratio distinguished MFS from the other patients at a threshold of 0.59. MFS patients were present in 24/58 (41.38%) patients with an AL/TCRP ratio of ≤0.59 and in 34/39 (87.18%) patients with an AL/TCRP ratio of >0.59. CONCLUSION: An AL/TCRP ratio of >0.59 is significantly associated with the risk of MFS. The AL/TCRP ratio should be measured as a promising marker for the prognosis of children MFS. Changes in the AL/TCRP ratio should be monitored over time.

4.
Ophthalmic Res ; 64(5): 811-819, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34034266

RESUMO

INTRODUCTION: Marfan syndrome (MFS) is characterized by ectopia lentis (EL) and elongated axial length (AL). The characteristics of AL in young patients with MFS and bilateral EL before the lens surgery are not fully understood. METHODS: This study reviewed MFS patients under 20 years old with bilateral EL from January 2015 to October 2020. The Z-scores were introduced in terms of the number of standard deviations from the mean of age-matched normative data. Using Z-scores, the distribution of AL and influence factors were evaluated. The correlations between AL and other biometrics were analyzed. RESULTS: We reviewed 183 patients and enrolled both eyes. The mean age was 8.44 ± 4.69 years. About 36% of the patients were children under 6 years old. The median AL increased from 23.16 mm under 5 years old to 26.20 mm in the 16-20 age group, and when plotted, the trend presented a logarithmic curvature (R2 = 0.145, p < 0.001). The median Z-AL score was 1.24. One-third of eyes had Z-score <0. About 20% of the patients had AL difference over 1 mm between the right and left eyes, and the right one had longer Z-AL scores (p = 0.013). The eye complicated with megalocornea (10, 7.04%) had larger Z-AL scores (4.72 ± 3.51 vs. 1.10 ± 2.25, p = 0.002). A positive correlation was found between Z-AL and Z-corneal curvature radius (r = 0.265, p < 0.001). CONCLUSION: Young patients with bilateral EL but small AL should not be excluded from MFS without systematic examination. The age-adjusted Z-score will facilitate further study of the individual variations in AL across different ages.

5.
Opt Lett ; 40(5): 744-7, 2015 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-25723422

RESUMO

A long-range surface plasmon resonance (LRSPR) sensor based on dielectric/silver-coated hollow fiber (HF) is proposed. It can detect the refractive index (RI) of sensed liquid filled in the hollow core of the sensor. A HF LRSPR sensor with 90-nm-thick silver layer and 260-nm-thick OC300 layer is fabricated. Experiments are taken to evaluate the performance of the sensor by measuring the transmission spectra. Theoretical analysis based on a ray model is also taken, and the results agree well with the experimental results. The proposed sensor has similar sensitivity but much smaller SPR dip width than the silver-coated HF SPR sensor. Thus figure of merit of the sensor is enhanced approximately five times. The stability of the sensor is also improved because the dielectric layer acts as a protection layer for the damageable silver layer.


Assuntos
Fibras Ópticas , Prata , Ressonância de Plasmônio de Superfície/instrumentação , Impedância Elétrica
6.
J Huazhong Univ Sci Technolog Med Sci ; 34(4): 548-553, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25135725

RESUMO

The changes of microRNA expression in rat hippocampus after traumatic brain injury (TBI) were explored. Adult SD rats received a single controlled cortical impact injury, and the ipsilateral hippocampus was harvested for the subsequent microarray assay at three time points after TBI: 1st day, 3rd day and 5th day, respectively. We characterized the microRNA expression profile in rat hippocampus using the microRNA microarray analysis, and further verified microarray results of miR-142-3p and miR-221 using quantitative real-time PCR. Totally 205 microRNAs were identified and up-/down-regulated more than 1.5 times. There were significant changes in 17 microRNAs at all three time points post-TBI. The quantitative real-time PCR results of miR-142-3p and miR-221 indicated good consistency with the results of the microarray method. MicroRNAs altered at different time points post-TBI. MiR-142-3p and miR-221 may be used as potentially biological markers for TBI assessment in forensic practice.


Assuntos
Lesões Encefálicas/metabolismo , Regulação da Expressão Gênica , Hipocampo/metabolismo , MicroRNAs/biossíntese , Animais , Biomarcadores/metabolismo , Lesões Encefálicas/patologia , Feminino , Genética Forense , Perfilação da Expressão Gênica , Hipocampo/patologia , Masculino , Ratos , Ratos Sprague-Dawley
7.
Opt Express ; 21(26): 32349-57, 2013 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-24514827

RESUMO

A new kind of surface plasmon resonance (SPR) sensor based on silver-coated hollow fiber (HF) structure for the detection of liquids with high refractive index (RI) is presented. Liquid sensed medium with high RI is filled in the hollow core of the HF and its RI can be detected by measuring the transmission spectra of the HF SPR sensor. The designed sensors with different silver thicknesses are fabricated and the transmission spectra for filled liquids with different RI are measured to investigate the performances of the sensors. Theoretical analysis is also carried out to evaluate the performance. The simulation results agree well with the experimental results. Factors that might affect sensitivity and detection accuracy of the sensor are discussed. The highest sensitivity achieved is 6,607 nm/RIU, which is comparable to the sensitivities of the other reported fiber SPR sensors.


Assuntos
Técnicas Biossensoriais/instrumentação , Tecnologia de Fibra Óptica/instrumentação , Refratometria/instrumentação , Soluções/análise , Soluções/química , Ressonância de Plasmônio de Superfície/instrumentação , Desenho de Equipamento , Análise de Falha de Equipamento , Porosidade
8.
Zhonghua Yan Ke Za Zhi ; 48(9): 829-35, 2012 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-23141580

RESUMO

OBJECTIVE: To explore the specific expression of HSV-tk gene and killing effects on ocular leading cells of the enhanced specific HSV-tk/GCV gene therapy system regulated by lens-specific promoter LEP503. METHODS: Experimental research. The enhanced specific HSV-tk/GCV gene system of two vectors were constructed (Lenti-LEP503-HSVtk-Cre and Lenti-HPGK-Loxp-EGFP-pA-Loxp-HSVtk). The lentiviral vectors were produced by transient transduction of transfering vectors, packaging vectors and enveloping vector into 293T cells. Virus was collected with ultracentrifugation and resuspended with 1 ml phosphate buffered saline and stored at -80°C. The HLEC and RPEC, NIH3T3, 293T cells were transduced with the enhanced specific HSV-tk gene system. The specific expressions of EGFP and HSV-tk were detected by fluorescence microscopy, flow cytometry and RT-PCR. The killing effects of HLEC and RPEC at the concentration of 20 mg/L GCV were assayed and compared by flow cytometry and CCK-8 kit. Difference of RPE cell viability among groups was evaluated by analysis of variance (ANOVA). RESULTS: Expression efficiency of EGFP in RPEC group was 62.3%, 68.3% in NIH3T3 group, 75.8% in 293T group, whereas 17.5% in HLEC group. There was higher expression of HSV-tk at mRNA level in HLEC group than that in RPEC group. The relative intensity of HSV-tk mRNA in HLEC group transduced with the enhanced specific HSV-tk gene system was 4.01, whereas 0.29 in RPEC group. At the concentration of 20 mg/L GCV after 72 hours, the percentage of apoptosis detected by the flow cytometry in HLEC group transduced by the enhanced specific HSV-tk gene system was 76.51%, and 2.44% in RPEC group. There was no significant difference in the RPE cell viability among the enhanced specific HSV-tk gene combination-RPE group, normal-RPE group and negative-RPE control group at the concentration of 20 mg/L GCV after 72 hours (MD(1) = -0.047, P = 0.671; MD(2) = 0.027, P = 0.912). CONCLUSIONS: The enhanced specific HSV-tk gene system express HSV-tk selectively in HLEC. At the concentration of 20 mg/L GCV, it is effective against the proliferation of HLEC in vitro, but has less kill effect on RPEC.


Assuntos
Proteínas de Ligação a DNA/genética , Técnicas de Transferência de Genes , Genes Transgênicos Suicidas/genética , Cristalino/citologia , Animais , Células Epiteliais , Fibroblastos/citologia , Expressão Gênica , Regulação da Expressão Gênica , Vetores Genéticos , Células HEK293 , Humanos , Camundongos , Células NIH 3T3 , Regiões Promotoras Genéticas , Epitélio Pigmentado da Retina/citologia , Simplexvirus/enzimologia , Timidina Quinase/genética
9.
PLoS One ; 7(8): e43173, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22916220

RESUMO

BACKGROUND: Cataract and geographic atrophy (GA, also called advanced "dry" age-related macular degeneration) are the two major causes of visual impairment in the developed world. The association between cataract surgery and the development of GA was controversial in previous studies. METHODS/PRINCIPAL FINDINGS: We performed a meta-analysis by pooling the current evidence in literature and found that cataract is associated with an increased risk of geographic atrophy with a summary odds ratio (OR) of 3.75 (95% CI: 95% CI: 1.84-7.62). However, cataract surgery is not associated with the risk of geographic atrophy (polled OR=3.23, 95% CI: 0.63-16.47). Further experiments were performed to analyze how the αA-crystallin, the major component of the lens, influences the development of GA in a mouse model. We found that theαA-crystallin mRNA and protein expression increased after oxidative stress induced by NaIO(3) in immunohistochemistry of retinal section and western blot of posterior eyecups. Both functional and histopathological evidence confirmed that GA is more severe in αA-crystallin knockout mice compared to wild-type mice. CONCLUSIONS: Therefore, αA-crystallin may protect against geographic atrophy. This study provides a better understanding of the relationship between cataract, cataract surgery, and GA.


Assuntos
Extração de Catarata , Catarata/metabolismo , Atrofia Geográfica/metabolismo , Atrofia Geográfica/cirurgia , Cadeia A de alfa-Cristalina/metabolismo , Animais , Western Blotting , Catarata/genética , Células Cultivadas , Eletrorretinografia , Atrofia Geográfica/genética , Humanos , Imuno-Histoquímica , Iodatos/farmacologia , Camundongos , Camundongos Knockout , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Cadeia A de alfa-Cristalina/genética
10.
Mol Vis ; 18: 2053-66, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22876133

RESUMO

PURPOSE: To establish a novel, targeted lentivirus-mediated LEP503-HSV-tk/GCV suicide gene therapy system combined with all trans-retinoic acid (ATRA) for the inhibition of human lens epithelial cell (HLEC) proliferation and treatment of posterior capsular opacification (PCO) after cataract surgery; to estimate the enhancement of the bystander effect by ATRA; and to explore the role of Connexin43 (Cx43) mediated gap junctional intercellular communication (GJIC) in the bystander effect of the HSV-K/GCV system. METHODS: A Lenti-LEP503-HSV-tk-EGFP vector was generated by cloning the lens-specific promoter LEP503 (lens specific promoter 503) from genomic DNA of HLECs by PCR. The vector was then inserted into the promoter-less vector from lentivirus-based (CMV)-HSV-tk-EGFP. The expressional specificity of the LEP503 promoter was assessed by investigating the expression of EGFP (enhanced green fluorescent protein) and HSV-tk (herpes simplex virus thymidine kinase) mRNA, both driven by Lenti-LEP503-HSV-tk-EGFP vector, by fluorescence microscopy, RT-PCR, flow cytometry, and western blot assays in HLECs, human adult retinal pigment epithelium cells (RPECs), human adult skin fibroblast cells (ASFCs), and Hela cells. Morphological changes were observed by fluorescence microscopy and cell viability was determined using the Cell Counting kit-8 Cell Proliferation (CCK-8) and MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays after Lenti-LEP503-HSV-tk/GCV system combined with ATRA treatment on HLECs. Flow cytometry, DNA fragmentation, and western blot assays were employed to analyze the mechanisms of bystander effects. RESULTS: The promoter LEP503-mediated HSV-tk was specifically expressed in HLECs, and ATRA dose-dependently strengthened the bystander effect following LEP503-mediated HSV-tk/GCV gene therapy against lens cells by upregulating the expression of the gap junction protein Cx43. CONCLUSIONS: The Lenti-LEP503-HSV-tk/GCV suicide gene therapy system, combined with ATRA as an adjuvant, may be a feasible supplementary method for PCO treatment that targets residual lens cells.


Assuntos
Efeito Espectador/efeitos dos fármacos , Conexina 43/genética , Proteínas de Ligação a DNA/genética , Cristalino/efeitos dos fármacos , Tretinoína/farmacologia , Efeito Espectador/genética , Proliferação de Células , Sobrevivência Celular/efeitos dos fármacos , Conexina 43/metabolismo , Proteínas de Ligação a DNA/metabolismo , Relação Dose-Resposta a Droga , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Junções Comunicantes/efeitos dos fármacos , Junções Comunicantes/genética , Junções Comunicantes/metabolismo , Expressão Gênica/efeitos dos fármacos , Genes Reporter , Genes Transgênicos Suicidas , Vetores Genéticos , Proteínas de Fluorescência Verde/genética , Células HeLa , Humanos , Cristalino/citologia , Cristalino/metabolismo , Lentivirus/genética , Regiões Promotoras Genéticas , Epitélio Pigmentado da Retina/citologia , Epitélio Pigmentado da Retina/efeitos dos fármacos , Epitélio Pigmentado da Retina/metabolismo
11.
Mol Vis ; 18: 74-80, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22262940

RESUMO

PURPOSE: To investigate whether apoptosis of human lens epithelial cells (HLECs) can be induced with the polyamidoamine (PAMAM)-mediated inhibition of bcl-2 (b-cell lymphoma 2) by small hairpin RNA (shRNA). METHODS: HLECs (SRA01/04) were transfected with the fifth generation of PAMAM (PAMAM G5) by bcl-2 shRNA. At 24, 48, and 72 h after transfection, the transfection rate was measured by flow cytometry. The transfection rates mediated by PAMAM and liposome were compared. The bcl-2 mRNA level was detected by real-time PCR. Whole cell protein was extracted and the bcl-2 protein level was detected by western blotting. The percentage of HLECs undergoing apoptosis was measured by Annexin V-FITC/PI staining. The nuclear morphology of HLECs was observed by staining with Hoechst 33258. The expression of cytochrome c and the activity of cleaved caspase-3 were analyzed by western blotting. RESULTS: At 24, 48, and 72 h after transfection, the rate of transfection of bcl-2 shRNA mediated by PAMAM was higher than in the liposome-mediated group (p<0.05). The mRNA and protein levels of bcl-2 were greatly downregulated. The percentage of HLECs undergoing apoptosis was greatly improved. Hoechst staining showed that bcl-2 shRNA transfected cells had a lower growth status with nuclear fragmentation. The expression of cytochrome c and the activity of cleaved caspase-3 was greatly improved (p<0.05). CONCLUSIONS: PAMAM-mediated bcl-2 shRNA can downregulate the expression of bcl-2 and induce the apoptosis of HLECs by engaging the mitochondrial pathway, including catalytic activation of the caspases.


Assuntos
Células Epiteliais/metabolismo , Cristalino/metabolismo , Poliaminas/química , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Mensageiro/biossíntese , RNA Interferente Pequeno/genética , Anexina A5/análise , Apoptose/genética , Western Blotting , Caspase 3/biossíntese , Linhagem Celular , Citocromos c/biossíntese , Células Epiteliais/citologia , Citometria de Fluxo , Regulação da Expressão Gênica , Inativação Gênica , Humanos , Cristalino/citologia , Lipossomos/química , Proteínas Proto-Oncogênicas c-bcl-2/genética , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais , Transfecção/métodos
12.
Ophthalmic Res ; 47(2): 87-97, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-21865765

RESUMO

AIM: To determine whether a diet containing excessive amounts of milk aggravates naphthalene-initiated cataracts in a common animal model of age-related human cataract. METHODS: Ninety Sprague-Dawley rats were fed a natural diet supplemented with either water (group A), normal amounts of milk (group B), excessive amounts of milk (group C), naphthalene plus water (group D), naphthalene plus normal amounts of milk (group E), naphthalene plus excessive amounts of milk (group F). Cataract development was monitored weekly using a slit lamp and lens gray value analysis. Concentrations of reactive oxygen species (ROS), reduced glutathione (GSH) and malondialdehyde (MDA) in rat lenses were measured to determine the role of oxidative stress in cataract induction. RESULTS: By week 4, the cortical gray value was significantly higher in group F than that in group D, and the cortical gray value was significantly higher in group D than in group A. However, by week 8, no significant differences were observed among groups C, F, B, E and A. ROS concentrations in lenses of rats of groups C and F were slightly higher than in those of groups B, E and A, but ROS concentrations in group F were significantly higher than in the other groups receiving naphthalene (i.e. groups D and E). GSH concentrations in group F were significantly lower than in the other groups. MDA concentrations in group F were significantly higher than in the other groups receiving naphthalene, indicating increased lipid peroxidation induced by naphthalene plus excessive intake of milk. CONCLUSIONS: Our results provide quantitative evidence that excessive intake of milk aggravates naphthalene-initiated cataracts, which is probably due to oxidative damage caused by increased ROS.


Assuntos
Catarata/induzido quimicamente , Leite/efeitos adversos , Estresse Oxidativo/fisiologia , Animais , Catarata/metabolismo , Catarata/fisiopatologia , Dieta , Modelos Animais de Doenças , Feminino , Glutationa/metabolismo , Cristalino/metabolismo , Masculino , Malondialdeído/metabolismo , Naftalenos , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Fatores de Risco
13.
Mol Vis ; 17: 291-9, 2011 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-21283526

RESUMO

PURPOSE: To establish a novel, targeted lentivirus-based HSV-tk (herpes simplex virus thymidine kinase)/GCV (ganciclovir) gene therapy system to inhibit lens epithelial cell proliferation for treatment of posterior capsular opacification (PCO) after cataract surgery. METHODS: An enhanced Cre recombinase (Cre/loxP) system with a lentiviral vector expressing Cre under the control of the lens-specific promoter LEP503 (Lenti-LEP503-HSVtk-Cre [LTKCRE]) was constructed, as well as another lentiviral vector containing a switching unit. The latter vector contains a stuffer sequence encoding EGFP (Lenti-hPGK-Loxp-EGFP-pA-Loxp-HSVtk [PGFPTK]) with a functional polyadenylation signal between two loxP sites, followed by the herpes simplex virus thymidine kinase (HSV-tk) gene, both under the control of the human phosphoglycerate kinase (hPGK) promoter. Expression of the downstream gene (HSV-tk) is activated by co-expression of Cre. Human lens epithelial cells (HLECs) or retinal pigmental epithelial cells (RPECs) were co-infected with LTKCRE and PGFPTK. The inhibitory effects on HLECs and RPECs infected by the enhanced specific lentiviral vector combination at the concentration of 20 µg/ml GCV were assayed and compared. RESULTS: The specific gene expression of Cre and HSV-tk in HLECs is activated by the LEP503 promoter. LTKCRE and PGFPTK co-infected HLECs, but not RPECs, expressed high levels of the HSV-tk protein. After 96 h of GCV treatment, the percentage of apoptotic HLECs infected by the enhanced specific lentiviral vector combination was 87.23%, whereas that of apoptotic RPECs was only 10.12%. Electron microscopy showed that GCV induced apoptosis and necrosis of the infected HLECs. CONCLUSIONS: The enhanced specific lentiviral vector combination selectively and effectively expressed HSV-tk in HLECs. A concentration of 20 µg/ml, GCV is effective against the proliferation of HLECs in vitro. This cell-type-specific gene therapy using a Cre/loxP lentivirus system may be a feasible treatment strategy to prevent PCO.


Assuntos
Extração de Catarata/métodos , Catarata/genética , Catarata/terapia , Ganciclovir/farmacologia , Terapia Genética/métodos , Simplexvirus/enzimologia , Timidina Quinase/genética , Antivirais/farmacologia , Apoptose , Linhagem Celular , Proliferação de Células , Citometria de Fluxo , Humanos , Integrases/metabolismo , Lentivirus/genética , Modelos Genéticos , Reação em Cadeia da Polimerase/métodos , Regiões Promotoras Genéticas
14.
Zhonghua Yan Ke Za Zhi ; 46(8): 679-85, 2010 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-21054990

RESUMO

OBJECTIVE: To evaluate the visual performance after implantation of the Tecnis ZM900 multifocal intraocular lens (IOL) (TMF) and the Restor SA60D3 multifocal IOL (Restor). METHODS: In a prospective study, TMF or Restor was implanted randomly in 73 patients (90 eyes). The following parameters were assessed 3 months after surgery: refraction, uncorrected and best corrected visual acuities (VA) for distance, intermediate, near and different contrast levels, reading ability, pupil size, wave-front error, defocus curve and position of IOL. Patient satisfaction (overall satisfaction, spectacle independence, photic phenomena) was assessed by a questionnaire. The chi-square test was applied to compare categorical variables and Mann-Whitney U test was used to compare the measured data. RESULTS: The uncorrected, best corrected and/or distance-corrected VA for distance, intermediate, near and different contrast levels did not show statistically significant differences between the two groups (P > 0.05). Near reading acuity and reading speed were better in TMF under low-light conditions (Z = -2.579, P = 0.009; Z = -5.244, P = 0.000). The curve of defocus showed that TMF had significantly better intermediate distance (at 50 cm) (Z = -5.300, P = 0.000) and worse near distance (from 25 to 28 cm) than those of Restor (Z = -3.745, P = 0.000; Z = -5.691, P = 0.000). Measurements under pupil diameter at 3.0 mm and 5.0 mm, ocular and intraocular Z (4, 0) were significantly lower (Z = -8.175, P = 0.000; Z = -5.210, P = 0.000 and Z = -4.453, P = 0.000; Z = -3.790, P = 0.000), the values of PSF Strehl Ratio and MTF AreaRatio A/D were significantly higher (Z = -3.047, P = 0.002; Z = -3.672, P = 0.008 and Z = -2.038, P = 0.042; Z = -2.579, P = 0.009) in TMF than those in Restor. On the questionnaire, there was no difference of overall satisfaction, spectacle independence and photic phenomena (P > 0.05). CONCLUSIONS: Implantation of the TMF and Restor offers excellent distant and near VA. Restor had better near VA than that of TMF based on the curve of depth, TMF had better VA at 50 cm-distance. Reading speed is faster in TMF. Compared to spherical Restor, TMF provides a better quality of vision due to a negative spherical aberration.


Assuntos
Catarata/terapia , Implante de Lente Intraocular , Lentes Intraoculares , Acuidade Visual , Sensibilidades de Contraste , Humanos , Estudos Prospectivos , Resultado do Tratamento
15.
Zhonghua Yan Ke Za Zhi ; 46(6): 518-24, 2010 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-21055197

RESUMO

OBJECTIVE: To evaluate the results of cataract surgery in myopia patients after laser in situ keratomileusis (LASIK) and to compare the predictability of various methods of intraocular lens (IOL) power calculation. METHOD: Seventeen cases (24 eyes) who had LASIK for myopia were divided into two group by with or without history of corneal power data. Corneal power was obtained by autokeratometry, corneal topography, Pentacam and IOL Master. The IOL power was calculated with the clinical history method, Feiz-Mannis formula, Feiz-Mannis method and other methods. Postoperative final refraction and the deviation of the final spherical equivalent (SEQ) from the refractive target were measured 3 month after the surgery. Two sample t-test, linear correlation and regression analysis, paired t-test and Bland-Altman method of agreement were used to analyze these data. RESULTS: In the group with history data, the mean corneal power was (43.28 ± 1.21) D and the mean SEQ was (-15.33 ± 4.36) D before the LASIK surgery. In the group without history data, the mean SEQ was (-10.11 ± 3.12) D. Before cataract surgery, the mean corneal power was (36.96 ± 2.07) D and (36.85 ± 1.40) D in these two groups. The mean arithmetic refractive prediction error after cataract surgery was (-0.66 ± 1.27) D and (-0.47 ± 0.82) D in these two groups, respectively. Data calculated by using Hamed-Wang-Koch method, Masket Formula, Koch/Maloney method, Shammar method and Pentacam ERK method were lower than the emmetropic IOL power. Data calculated by using Feiz-Mannis Formula, Latkany Method, Savini method, Armberri Double K method were overestimated. The mean arithmetic errors of clinic history method, Corneal Passby Method and Haigis-L Formula were not significantly different from the predict refraction (P = 0.364, 0.318 and 0.069; t = 0.956, -1.057 and -1.911, respectively). There was strong correlation between the value calculated by using Feiz-Mannis Method or Haigis-L Formula and the true power (r = 0.921, 0.915; P = 0.000 and 0.000, respectively). But none of the values calculated by these method could fully agree with the true value. CONCLUSIONS: IOL power should be calculated accurately to avoid undercorrection. We recommend the combination of clinical history method, Feiz-Mannis Method, Corneal Passby Method and Haigis-L Formula for the calculation of IOL power.


Assuntos
Catarata/terapia , Miopia/cirurgia , Procedimentos Cirúrgicos Refrativos/métodos , Adulto , Córnea/cirurgia , Humanos , Implante de Lente Intraocular , Lentes Intraoculares , Pessoa de Meia-Idade , Refração Ocular
16.
Graefes Arch Clin Exp Ophthalmol ; 248(5): 681-6, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20162296

RESUMO

BACKGROUND: To present the visual results and the complications of primary intraocular lens (IOL) implantation in infants aged 6 to 12 months between January 2002 and July 2007. METHODS: A total of 26 consecutive eyes, of 16 infants with cataract aged 6 to 12 months, were reviewed in the study. All patients had cataract extraction with anterior and posterior capsulorrhexis combined with anterior vitrectomy and primary hydrophobic acrylic IOL implantation. Six infants (six eyes) had unilateral congenital cataract and ten (20 eyes), bilateral cataract. Visual acuity and complications were recorded throughout the 46.4-month mean follow-up (range 22 to 79 months). RESULTS: All eyes had primary IOL implantation. The mean best-corrected visual acuity (logMAR) was 0.98 +/- 0.18,0.50 +/- 0.14 and 0.61 +/- 0.25 for unilateral, bilateral and all eyes respectively at the last follow-up. IOLs were implanted in the capsular bag of 25 eyes (96.2%) and in the sulcus of the remaining one eye (3.8%). Seven eyes (26.9%) developed visual axis opacification (VAO), and four eyes required secondary pars plana vitrectomy (PPV). IOL opacification occurred in one eye 54 months after implantation. Late onset open-angle glaucoma developed in one eye, and required trabeculectomy surgery. The predictors of good best-corrected visual acuity (BCVA) included partial cataract, bilateral cataract, absence of strabismus or nystagmus, and good amblyopic treatment. The greatest annual myopic change (5.15 +/- 2.08 D) was observed during the first 12 months after surgery. In unilateral cases, there was no significant difference in the axial length between the cataractous eye and the fellow normal eye both at the time of surgery (P = 0.891) and final follow-up (P = 0.693). CONCLUSIONS: Primary IOL implantation was safe and effective for infantile cataract surgery. Total or unilateral cataract, nystagmus or strabismus, and inadequate amblyopic therapy were predictors of poor BCVA. Significant myopic shifts occurred especially in infants in the first year of surgery. The pseudophakic eye had a similar growth rate, as measured by axial length, to that of the fellow normal eye, in unilateral cases.


Assuntos
Extração de Catarata , Catarata/congênito , Complicações Intraoperatórias , Implante de Lente Intraocular , Complicações Pós-Operatórias , Acuidade Visual/fisiologia , Capsulorrexe , Seguimentos , Humanos , Lactente , Estudos Retrospectivos , Transtornos da Visão/reabilitação
17.
Zhonghua Yan Ke Za Zhi ; 45(7): 636-40, 2009 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-19957693

RESUMO

OBJECTIVE: To investigate whether apoptosis of human lens epithelial cells (HLEC) can be induced by inhibition of bcl-2 shRNA. METHODS: It was an experimental study. Two pairs of oligonucleotides were synthesized and inserted into plasmid PGCsi to generate shRNA eukaryotic expression vectors, named P1 and P2. At 48 hours after transfection in HLEC (SRA01/04) with Lipofectamine 2000, the whole cell protein was extracted and detected by Western blot. The bcl-2 mRNA level was detected by real-time PCR. The percentage of HLECs undergoing apoptosis was measured by Annexin V-PI staining. The activity of caspase-3 was analyzed by Western blotting. RESULTS: The shRNA eukaryotic expression vectors were constructed successfully. At 48 hours after transfection, the rate of transfection of P1 and P2 was about 44.1% and 47.2% respectively. The protein and mRNA level of bcl-2 was 0.435 and 0.476, greatly downregulated (F = 1672.4, P < 0.05). The percentage of HLEC undergoing apoptosis was 42.3% and 45.4%, greatly improved (F = 1756.2, P < 0.05). The activity of caspase-3 was greatly improved. CONCLUSION: P1 and P2 can both down-regulate the expression of bcl-2, and induce the apoptosis of HLEC.


Assuntos
Apoptose , Células Epiteliais/citologia , Genes bcl-2 , Cristalino/citologia , Interferência de RNA , Catarata , Linhagem Celular , Regulação para Baixo , Células Epiteliais/metabolismo , Humanos , Cristalino/metabolismo , RNA Interferente Pequeno/metabolismo , Transfecção
18.
Zhonghua Yan Ke Za Zhi ; 44(10): 902-5, 2008 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-19176118

RESUMO

OBJECTIVE: To explore the expression of telomerase activity of the lens epithelial cells in posterior capsule opacification of rabbits. METHODS: Clear corneal tunnel phacoemulsification was performed in both eyes of 11 New Zealand rabbits. After the model of posterior capsule opacification succeed in all eyes. Then, telomerase activity of the lens epithelial cells in the equator and posterior capsule opacification of 20 eyes was detected with TRAP-ELISA and TRAP-PAGE techniques. RESULTS: HepG2 cells were used as positive controls. Telomerase activity was detected in the lens epithelial cells in the equator capsule and posterior capsule opacification in rabbits, which showed several dim gradient stripped electrophoresis pattern. A450 - 690 value of telomerase activity in the equator capsule and posterior capsule opacification was 0.85 +/- 0.23 and 0.67 +/- 0.19, respectively, indicating statistically significant difference (t = 2.526, 0.021; P < 0.05). CONCLUSIONS: Telomerase activity exists in the lens epithelial cells of posterior capsule opacification in rabbits. The telomerase activity in this area is lower than that in the equator capsule.


Assuntos
Catarata/metabolismo , Células Epiteliais/metabolismo , Cristalino/metabolismo , Telomerase/metabolismo , Animais , Catarata/patologia , Feminino , Células Hep G2 , Humanos , Cristalino/citologia , Cristalino/patologia , Masculino , Coelhos
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