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1.
Article in Chinese | WPRIM | ID: wpr-883358

ABSTRACT

RNA-seq is a transcriptome analysis method using deep sequencing technology, which uses high-throughput next-generation sequencing technology to investigate, characterize and quantify transcriptome.It can quickly and comprehensively detect almost all transcripts and gene sequences of specific cells in specific tissue of a species, and has been widely used in basic research, clinical diagnosis and drug development and other fields.Its measurement accuracy has been confirmed comparable to microarray and quantitative polymerase chain reaction by several studies.RNA-seq technology can accurately detect mRNA and non-coding RNA which play important roles in the pathogenesis of diabetic retinopathy (DR). Using this technology to investigate the gene regulation and molecular mechanism of DR is helpful to find biomarkers for early diagnosis and prognosis of DR, to comprehensively and systematically study and analyze the molecular mechanism of specific biological process, and to find new therapeutic targets.The application of RNA-seq will be of great significance for the basic research and clinical treatment of DR.In this article, RNA-seq technology was comprehensively and systematically expounded from three aspects, which were the advantages of RNA-seq technology, the selection of sequencing platform as well as library preparation and data analysis.The progress of this technology in DR research was summarized and analyzed.

2.
Article in Chinese | WPRIM | ID: wpr-865325

ABSTRACT

With the development of intraocular injection of anti-vascular endothelial growth factor (VEGF) drugs, early treatment of polypoidal choroidal vasculopathy (PCV) has more understanding and development.Once PCV combined with vitreous hemorrhage, treatment becomes much more difficult.The causes of vitreous hemorrhage in PCV, treatment of PCV with vitreous hemorrhage by different surgical methods, including simple vitrectomy, vitreous gas injection, intraoperative combined with vitrectomy and PDT, vitrectomy combined with CF 6, retinal pigment epithelium transplantation and other vitrectomy, as well as common operation complications, postoperative recurrence and prognosis are summarized in this article.

3.
Article in Chinese | WPRIM | ID: wpr-871844

ABSTRACT

Objective:To observe the changes in refractive status of eyes with idiopathic macular hole (IMH) after vitrectomy and phacoemulsification and IOL implantation (combined surgery).Methods:A retrospective clinical study. From January 2016 to June 2019, 51patients (56 eyes) of IMH who underwent combined surgery at the Tianjin Medical University Eye Hospital. were included in the study. Among them, there were 17 males and 34 females with the average age of 66.79±4.33 years. All the affected eyes underwent BCVA, retinoscopy and axial length (AL) measurement. The IOL power was calculated according to the SRK-T formula and the refractive power (predicted value) was predicted. The average BCVA of the affected eye was 0.20±0.13. The average anterior chamber depth was 2.89±0.28 mm. The average △corneal astigmatism was 0.73±0.43 D, the average AL was 22.92±0.70 mm, the average predicted refractive power was 0.10±0.66 D. All the affected eyes underwent standard transciliary flat part three-channel 25G combined surgery. Six months after the operation, the actual value (actual value) of the diopter after the operation was measured with the same equipment and method before the operation. Paired t test was used to compare the difference between the predicted value and the actual value. Results:Six months after the operation, the actual value of the refractive power was -0.19±0.64 D. Compared with the pre-operative refractive power, the difference was not statistically significant ( t=1.665, P=0.102). The difference between the actual value and the predicted value was -0.33± 0.81 D. Conclusions:The refractive status of the IMH eye undergoes myopia drift after combined surgery. The preoperative IOL power budget can be appropriately reserved for +0.3 D hyperopia.

4.
Article in Chinese | WPRIM | ID: wpr-871754

ABSTRACT

Accurate collection and preservation of vitreous and retina-related tissue specimens is the basis for clinical diagnosis and rigorous basic research. The clinical uses of vitreous specimens include microbial culture, cytological detection, detection of degenerative diseases, PCR analysis, and cytological detection of cell morphology. The experimental research uses include DNA gene analysis, protein quantitative analysis, metabolite examination, RNA content quantitative analysis, cytokine determination and so on. Retinal specimens collecting was mainly used for PCR analysis of retinal proliferative membrane, immunohistochemical staining, immunofluorescence examination, microvascular density evaluation, cell isolation and culture, etc. Understanding the collection of vitreoretinal surgical specimens and the application of relevant detection techniques and materials can provide a more comprehensive idea for the diagnosis of vitreoretinal diseases and a broader reference for the related basic research.

5.
Article in Chinese | WPRIM | ID: wpr-871729

ABSTRACT

Objective:To observe the changes of follistatin-like protein 1 (FSTL1) in serum of patients with proliferative diabetic retinopathy (PDR).Methods:Twenty PDR patients confirmed by clinical examination and 20 normal people were included in the study. Human retinal vascular endothelial cells (HRCEC) were divided into HRCEC blank control group, 3 h hypoxia group, 6 h hypoxia group. Human umbilical vein endothelial cell (HUVEC) were divided into HUVEC blank control group, 3h hypoxia group, 6h hypoxia group. Real-time quantitative PCR (RT-PCR) and ELISA were used to determine the expression of FSTL1, TGF-β, VEGF, connective tissue growth factor (CTGF) mRNA and protein in peripheral blood and cells of all groups from all subjects.Results:The expressions of FSTL1, TGF-β1, CTGF, VEGF mRNA in blood samples of patients with PDR were 1.79±0.58, 0.97±0.21, 1.85±0.69 and 1.38±0.44. The expressions of FSTL1, TGF-β1 protein were 1.19±0.50, 0.71±0.24 ng/ml and 734.03±116.45, 649.36±44.23 ng/L. Compared with normal people, the differences were statistically significant ( tmRNA=0.90, 0.21, 2.85, 1.77; P=0.00, 0.00, 0.04, 0.02. tprotein=1.88, 7.68; P=0.00, 0.02). The cell viability of HRCEC cells in the 3 h hypoxia group and the 6 h hypoxia group were 0.66±0.05 and 0.64±0.04, respectively. Compared with the blank control group, the difference was statistically significant ( F=13.02, P=0.00). The cell viability of HUVEC cells in the 3 h hypoxia group and the 6 h hypoxia group were 0.63±0.06 and 0.68±0.06, respectively. Compared with the blank control group, the difference was statistically significant ( F=26.52, P=0.00). Comparison of FSTL1, TGF-β1, CTGF, and VEGF mRNA expression in HRCEC blank control group and 3 h hypoxia group, the differences were statistically significant ( F=14.75, 44.93, 85.54, 6.23; P=0.01, 0.00, 0.00, 0.03). Compared with the HRCEC blank control and 3 h hypoxia group, the expressions of FSTL1 and TGF-β1 protein were statistically significant ( P<0.05). There was a statistically significant difference in TGF-β1 protein expression in the hypoxic 6 h group ( P=0.03) and no significant difference in FSTL1 protein expression ( P=0.68). Comparison of FSTL1, TGF-β1, CTGF, and VEGF mRNA expression in HUVEC blank control group and 3h hypoxia group, the differences were statistically significant ( F=19.08, 25.12, 22.89, 13.07; P=0.00, 0.00, 0.00, 0.01). Immunofluorescence staining results showed that FSTL1, TGF-β1, CTGF, and VEGF proteins were positively expressed in cells in the 3h hypoxia and 6h hypoxia groups. Conclusion:The expression of FSTL1 gene and protein in serum of PDR patients was significantly higher than that of normal people.

6.
Article in Chinese | WPRIM | ID: wpr-746192

ABSTRACT

Objective To observe the effect ofpolypyramidine tract binding protein-associated splicing factor (PSF) towards advanced glycation end products (AGEs) induced the apoptosis of Müller cells in vitro.Methods Experimental study.Müller cells were cultured and divided into groups according to the project design,plasmid enhanced green fluorescent protein-PSF were transfected into the cells to achieve the overexpression of PSF Müller cells in vitro,then cells were exposed to AGEs and the Morphological changes were observed by HE staining and Hoechst 33258 staining while the survival rate of cells were detected by MTT assay.The effects of PSF on AGEs-induced Müller apoptosis was measured by Cell Death Detection ELISA kit.Meanwhile,2',7'-diehlorofluorescin diaeetate staining was performed to monitor the protective effects of PSF on AGEs-induced Müller cells ROS.Results The morphology of cells in normal group was full and the cytoplasm staining was uniform.In N+AGEs group and Vec+AGEs group,cell volume decreased,cytoplasm was dense and concentrated,and eosinophilic staining was enhanced.The cell morphology of PSF+AGEs group was still full,with uniform cytoplasm staining and uniform nucleus staining.The viability of N+AGEs group,Vec+AGEs group and PSF+AGEs group were 0.42±0.11,0.35±0.12 and 0.68±0.12.The apoptosis values were 1.08 ± 0.16,0.96± 0.20 and 0.44± 0.08.The intracellular ROS levels were 28 833.67± 3 550.06,28 356.67±4 854.81,186 163.00±382.54.Compared with N+AGEs group and Vec+AGEs group,the cell viability of PSF+AGEs group was significantly improved (F=20.65,P=0.000),ce11 apoptosis value (F=43.43,P=0.000) and intracellular ROS level (F=1 8.86,P=0.000).Conclusion PSF overexpression play a protective role in AGEs-induced apoptosis by inhibiting the production of ROS in Müiller cells.

7.
Article in Chinese | WPRIM | ID: wpr-711972

ABSTRACT

Objective To construct the connective tissue growth factor (CTGF) recombinant interference vector (shRNA) and observe its inhibitory effect on the expression of endogenous CTGF in retinal vascular endothelial cells.Methods The human CTGF shRNA was constructed and the high-titer CTGF shRNA lentivirus particles was acquired via three-plasmid lentivirus packaging system to infect retinal vascular endothelial cells.The optimal multiplicity and onset time of lentivirus infection were identified by tracing down the red florescent protein in interference vector.The cells were classified into three groups:blank control group,infection control group and CTGF knockdown group.The differences in cells migrating ability was observed through Transwell allay.The mRNA and protein expression of CTGF,fibronectin,a-smooth muscle actin (α-SMA) and collagen Ⅰ (Col Ⅰ) were quantified through real-time PCR testing and Western blot system.Data between the three groups were examined via one-way analysis of variance.Results The result showed that an optimal multiplicity of 20 and onset time of 72 hours were the requirements to optimize lentivirus infection.Transwell allay result showed a contrast in the number of migrated cells in the CTGF knockdown group and that in the blank control group and infection control group (F=20.64,P=0.002).Real-time PCR testing showed a contrast in related gene expression (CTGF,fibronectin,α-SMA and Col Ⅰ) in the CTGF knocked-down group and that in the blank control group and infection control group (F=128.83,124.44,144.76,1 374.44;P=0.000,0.000,0.000,0.000).Western blot system showed the statistical significance of the contrasted number of related protein expression (CTGF,fibronectin,α-SMA and Col Ⅰ) in the knockdown group and that in the blank control group (F=22.55,41.60,25.73,161.68;P=0.002,0.000,0.001,0.000).Conclusion The success in producing CTGF shRNA lentivirus particle suggests that CTGF shRNA lentivirus can effectively knock down CTGF expression.

8.
Article in Chinese | WPRIM | ID: wpr-711957

ABSTRACT

Intravitreal injections of anti-vascular endothelial growth factor (anti-VEGF) drugs,including monoclonal antibodies (such as bevacizumab and ranibizumab) and fusion protein agents (such as aflibercept and conbercept) have been proven to be effective in the treatment of wet age-related macular degeneration (wAMD).However,there are still some patients with poor efficacy,such as no response to initial treatment or poor response,and even relapse during the course of treatment.In view of the different targets and molecular characteristics of anti-VEGF drugs,the switch of anti-VEGF drugs and the adjustment of delivery pattern,dosages and intervals have been the strategies to cope with the poor efficacy in clinic.However,there are some differences in the results of current studies.Overall,the recovery of retinal anatomical outcome achieves more benefits,and it is relatively difficult to improve visual acuity.To determine which regimen would get the biggest benefits,a large number of randomized controlled clinical trials and long study period will be needed.

9.
Article in Chinese | WPRIM | ID: wpr-711918

ABSTRACT

Objective To observe RNA-Seq analysis of gene expression profiling in human retinal vascular endothelial cells after anti-vascular endothecial growth factor (VEGF) treatment.Methods Cultured the retinal vascular endothelial cells in vitro and logarithmic growth phase cells were used for experiments.The cells were divided into VEGF group and VEGF combined with anti-VEGF drugs group.The VEGF group cells were treated with 50 ng/ml VEGF for 72 h to simulate the high VEGF survival conditions of vascular endothelial cells in diabetic retinopathy.VEGF combined with anti-VEGF drug group cells was treated with 50 ng/ml VEGF and 2.5 μg/ml anti-VEGF drugs for 72 h to imitate the microenvironment of cells following the anti-VEGF drugs treatment,and whole transcriptome sequencing approach was applied to the above two groups of cells through RNA-Seq.Now with biological big data obtained as a basis,to analyze the differentially expressed genes (DEGs).And through enrichment analysis to explain the differential functions of DEGs and their signal pathways.Results The gene expression profiles of the two groups of cells were obtained.Through analysis,328 DEGs were found,including 194 upregulated and 133 downregulated ones.The functions of DEGs were influenced by regulations over molecular biological process,cellular energy metabolism and protein synthesis,etc.Among these genes,SI,PRX and HPGD were related to protein synthesis,BIRCT to cellular apoptosis,and ABLIM 1 and CRB2 to retinal development,and ABCG 1,ABCA9 and ABCA12 were associated with the cholesterol of macrophage and the transfer of phospholipid.GO enrichment analysis showed that DEGs mainly act in three ways:regulating biological behavior,organizing cellular component and performing molecular function.Pathway enrichment analysis showed that gene expressions of the two cell groups were differentiated in ECM receptor pathway,and Notch,mitogen-activated protein kinase,transforming growth factor (TGF)-β and Wnt signal pathways.Among them,the gene expression in TGF-β signal pathway attracts most attention,where the DEGs,such as CAMK2B,COL3A1,CYGB,PTGER2 and HS6ST2,among others,were closely related to fibrosis process.Conclusion The anti-VEGF drugs may enhance the expression ofCAMK2B,COL3A1,CYGB,PTGER2 and others genes related to TGF-β signal pathway and aggravate retinal fibrosis disease.

10.
Article in Chinese | WPRIM | ID: wpr-711894

ABSTRACT

Objective To observe the effect ofpolypyramidine tract binding protein-associated splicing factor (PSF) on hydrogen peroxide (H2O2) induced apoptosis of retinal pigment epithelial (RPE) cells in vitro.Methods RPE cells were cultured and divided into a normal group,normal+H2O2 group,Vec+H2O2 group,PSF+H2O2 group according to the experimental design.Overexpression of PSF in RPE cells were achieved by pEGFP-PSF plasmid transient transfection into RPE cells,then RPE cells were exposed to H2O2.The morphological changes were observed by hematoxylin-eosin (HE) staining and Live/Dead staining while the survival rate of cells was detected by MTT assay.The effect of PSF on H2O2-induced RPE apoptosis was analyzed by Cell Death Detection ELISA kit.Meanwhile,intracellular reactive oxygen species (ROS) level was detected by using DCFH-DA method.Results Overexpression of PSF could effectively alleviate the morphological changes induced by H2O2 stimulation shown by HE staining,and effectively reduce dead cells number shown by Live/Dead staining.After H2O2 stimulation,the survival rate,apoptosis rate and ROS production level in PSF overexpression group were 0.68± 0.12,0.44± 0.08 and 18 616± 3 382.54 respectively,showing significant difference in comparison with the vector plasmid group and normal group (P<0.05).Conclusion PSF overexpression plays a protective role in H2O2-induced apoptosis by inhibiting the production of ROS in RPE cells.

11.
Article in Chinese | WPRIM | ID: wpr-511135

ABSTRACT

Objective To evaluation clinical efficacy and complications characteristics of vitrectomy on type 1 diabetes of diabetic retinopathy.Methods This was a retrospective review study.From January 2010 to January 2016,18 cases (28 eyes) under the age of 30 diagnosed as type 1 diabetes of diabetic retinopathy were treated in our hospital.All the patients were performed vitrectomy.The postoperative visual acuity and complications in and after operation were observed.Results The follow-upperiod was 6-72 months with the average of 43.0 months.Postoperative visual acuity improved or remained unchanged in 23 eyes (82.1%),and decreased in 5 eyes(17.9%).Our primary anatomic success rate was 92.3%.Complication in operation was iatrogenic retinal tear in 3 eyes.Postoperative complications consisted of anterior chamber fibrin exudation in 9 eyes,hyphema in 5 eyes,recurrent vitreous hemorrhage in 2 eyes,retinal detachment in 3 eyes,rubeosis of iris in 5 eyes and neovascular glaucoma in 4 eye.Conclusion For young patients of type 1 proliferative diabetic retinopathy,vitrectomy can restore retinal anatomical structure and function.Panrentinal photocoagulation and preoperative anti-VEGF injections in vitreous is necessary with serious neovascularization-related complications.

12.
Tianjin Medical Journal ; (12): 657-659, 2017.
Article in Chinese | WPRIM | ID: wpr-612359

ABSTRACT

The incidence of high myopia has increased yearly, which has been an area of focus for investigation. The complications of high myopia are contributing factors to blindness,such as posterior staphyloma, chorio-retinal atrophy, choroidal neovascularization, optic nerve head and macular defects. The study of histological changes helps clarify the mechanism. With the development of modern technology such as spectral-domain optical coherence tomography and enhanced depth imaging-technique, histological changes in high myopia can be measured accurately. The purpose of this review is to provide an update on various pathologic changes, including in sclera, Choroid/Bruch's membrane, optic nerve, vitreous and retina.

13.
Article in Chinese | WPRIM | ID: wpr-381455

ABSTRACT

Objective To observe the therapeutic efficacy and postoperative complications of combined surgery and sequential surgery in treating proliferative diabetic retinopathy (PDR) and cataract.Methods The cilinical data of 59 patients (66 eyes) with PDR were retrospectively analyzed.The patients were divided into combined-surgery group and sequential-surgery group.Combined surgery was performed on patients with obvious lens opacity which was an obstacle to the ocular fundus surgery,while sequential surgery was performed on the patients with transparent lens or lens with light opacity on which ocular fundus surgery could be performed.Lens excision in cataractopoiesis could only be performed when the state of ocular fundus was stable and cataractopoiesis was the main cause of vision damage.A total of 28 patients (32 eyes) in combined-surgery group underwent vitrectomy combined with phacoemulsification and IOL implantation;31 patients (34 eyes) in sequential-surgery group underwent vitrectomy,lens excision and IOL implantation.Corrected visual acuity (BCVA) and complications were observed.The mean follow-up period was (25±8.5) months.The measurement data were analyzed with t test and enumeration data with 2 test.Results In combined-surgery group,visual acuity improvement was achieved in 27 eyes (84.4%),remained unchanged in 2 eyes (6.3%) and decreased in 3 eyes (9.4 %).In sequential-surgery group,visual acuity improved in 26 eyes (76.5%),remained unchanged in 2 eyes(5.8%)and decreased in 6 eyes (17.7%.).However,anterior chamber fibrin exudation occurred in 4 eyes in the combined-surgery group and no eyes in the sequential-surgery group;compared with each other,the difference is statistically significant (χ2=4.524,P=0.033).Conclusion Combined surgery and sequential surgery are all safe and effective on treating PDR and postoperative complications have no obvious correlation with the surgery procedure.

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