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Objective:To investigate the differential expression profile of circular RNA (circRNA) in high glucose-cultured human retinal vascular endothelial cells (HRVECs).Methods:HRVECs were divided into high glucose group, normal control group and hypertonic control group, and were cultured in 25 mmol/L glucose medium, 5.5 mmol/L glucose medium and 19.5 mmol/L mannitol+ 5.5 mmol/L glucose medium for 24 hours accordingly.The differentially expressed circRNA molecules between high glucose group and normal control group were screened by circRNA microarray analysis.The expression of the most significant differentially expressed circRNAs in different groups was verified by real-time quantitative PCR.The possible microRNA (miRNA) targets were analyzed through the Circular RNA Interactome database.Results:It was found that 448 circRNAs were differentially expressed (FC≥1.5 or FC≤0.67, P<0.05) in high glucose-cultured HRVECs, among which 182 were up-regulated and 266 were down-regulated.The top 3 significantly up-regulated circRNAs were hsa_circ_0002938, hsa_circ_0008036, and hsa_circ_0001946, and the top 3 significantly down-regulated circRNAs were hsa_circ_0035277, hsa_circ_0008344, and hsa_circ_0001874.Compared with normal control group and hypertonic control group, the relative expressions of top 3 up-regulated circRNAs were significantly enhanced and the relative expressions of top 3 down-regulated circRNAs were significantly reduced in high glucose group, showing statistically significant differences (all at P<0.05).No significant difference was found in the differentially expressed circRNAs between normal control group and hypertonic control group (all at P>0.05). Conclusions:CircRNAs are differentially expressed in high glucose-cultured HRVECs, and the differentially expressed circRNAs may be involved in the regulatory mechanism of diabetic retinopathy.
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Objective To investigate the nature of the suprachoroidal fluid by detecting the concentration of total protein (TP), lactate dehydrogenase (LDH), albumin (ALB), total cholesterol (CHOL), total bilirubin (TBIL) in suprachoroidal liquid of patients who have rhegmatogenous retinal detachment with choroid detachment (RRDCD). Methods Eighteen RRDCD patients (18 eyes) who underwent vitrectomy were enrolled in this study. There were 10 males (10 eyes) and 8 females (8 eyes), 8 right eyes and 10 left eyes. There were 8 patients with age of ≤55 years, 10 patients with age of >55 years. There were 7 patients with duration of≤30 days, 11 patients with duration of >30 days. There were 7 eyes with diopters of ≥-6.0 D, 11 eyes with diopters of <-6.0 D. There were 11 eyes with class C proliferative vitreoretinopathy (PVR), 7 eyes with class D PVR. Suprachoroidal fluid samples were collected from all the patients, and took preoperative serum samples as RRDCD group. Ten serum samples of normal people were set as control group. The concentration of TP, LDH, ALB, CHOL, TBIL in all the subjects were measured. The properties of the suprachoroidal fluid were identified by Light standard and concentration standard of ALB, CHOL, TBIL. Results There was no difference on the concentration of TP, LDH, ALB, CHOL, TBIL from suprachoroidal fluid samples in the patients with different age, sex, eyes, diopter, PVR grade (P>0.05). There was no difference on the concentration of TP, LDH, ALB, CHOL, TBIL from preoperative serum samples in the patients between RRDCD group and control group (P>0.05). There was no difference on the concentration of ALB and CHOL from suprachoroidal fluid samples and preoperative serum samples in the RRDCD patients (P>0.05), but there were significant differences on the concentration of TP, LDH, TBIL (P<0.05). According to the Light standard, there were 17 cases of exudates and 1 case of transudate. According to the concentration standard of ALB, CHOL and TBIL, there were 14, 18, and 16 cases of exudates, and 4, 0, and 2 cases of transudate, respectively. There was no difference on the identification result of Light standard and concentration standard of ALB, CHOL, TBIL (χ2=2.090, 1.029, 0.364;P>0.05). Conclusion The suprachoroidal fluid of RRDCD patients composed of TP, LDH, CHOL and TBIL. The suprachoroidal fluid is more likely to be exudate.
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Objective To investigate the nature of the suprachoroidal fluid by detecting the concentration of total protein (TP), lactate dehydrogenase (LDH), albumin (ALB), total cholesterol (CHOL), total bilirubin (TBIL) in suprachoroidal liquid of patients who have rhegmatogenous retinal detachment with choroid detachment (RRDCD). Methods Eighteen RRDCD patients (18 eyes) who underwent vitrectomy were enrolled in this study. There were 10 males (10 eyes) and 8 females (8 eyes), 8 right eyes and 10 left eyes. There were 8 patients with age of ≤55 years, 10 patients with age of >55 years. There were 7 patients with duration of≤30 days, 11 patients with duration of >30 days. There were 7 eyes with diopters of ≥-6.0 D, 11 eyes with diopters of <-6.0 D. There were 11 eyes with class C proliferative vitreoretinopathy (PVR), 7 eyes with class D PVR. Suprachoroidal fluid samples were collected from all the patients, and took preoperative serum samples as RRDCD group. Ten serum samples of normal people were set as control group. The concentration of TP, LDH, ALB, CHOL, TBIL in all the subjects were measured. The properties of the suprachoroidal fluid were identified by Light standard and concentration standard of ALB, CHOL, TBIL. Results There was no difference on the concentration of TP, LDH, ALB, CHOL, TBIL from suprachoroidal fluid samples in the patients with different age, sex, eyes, diopter, PVR grade (P>0.05). There was no difference on the concentration of TP, LDH, ALB, CHOL, TBIL from preoperative serum samples in the patients between RRDCD group and control group (P>0.05). There was no difference on the concentration of ALB and CHOL from suprachoroidal fluid samples and preoperative serum samples in the RRDCD patients (P>0.05), but there were significant differences on the concentration of TP, LDH, TBIL (P<0.05). According to the Light standard, there were 17 cases of exudates and 1 case of transudate. According to the concentration standard of ALB, CHOL and TBIL, there were 14, 18, and 16 cases of exudates, and 4, 0, and 2 cases of transudate, respectively. There was no difference on the identification result of Light standard and concentration standard of ALB, CHOL, TBIL (χ2=2.090, 1.029, 0.364;P>0.05). Conclusion The suprachoroidal fluid of RRDCD patients composed of TP, LDH, CHOL and TBIL. The suprachoroidal fluid is more likely to be exudate.
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Objective To observe the effect of panretinal photocoagulation (PRP) on the expression of cyclooxygenase-2 (COX-2),vascular endothelial cell growth factor (VEGF) in epiretinal membrane of proliferative diabetic retinopathy (PDR).Methods A total of 35 patients (35 eyes) with PDR and underwent plana vitrectomy were enrolled in this study.The patients were divided into non-PRP group (19 patients,19 eyes) and PRP group (16 patients,16 eyes) depends on if they had received PRP before surgery.The epiretinal membranes stripped during operation were collected for pathological examination.The histopathological features was observed by haematoxylin and eosin stain.The expression of CD34,COX-2 and VEGF,and microvessel density (MVD) were measured by immunohistochemistry method.Results Many new dispersed capillary blood vessels were found in the thick epiretinal membranes of nonPRP group,while scattered small blood vessels were found in the relatively thin epiretinal membranes of PRP group.MVD value was (7.42± 1.39) in the non-PRP group and (4.56± 1.22) in the PRP group,which was lower than the non-PRP group (t=6.41,P＜0.01).The expression of CD34,COX-2 and VEGF in the tissues of epiretinal membrane in PRP group were obviously lower than the non-PRP group (t=6.147,5.944,7.445;P＜0.01).Conclusion PRP can effectively inhibit the expression of COX-2 and VEGF in epiretinal membrane of PRP patients.
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Background Studies showed that some members of matrix metalloproteinases (MMPs) play an important role in the pathogenesis of diabetic retinopathy (DR).However,whether MMPs inhibitor can deter blood retinal barrier (BRB) from damage is below understood.Objective This study was to investigate the effects of GM6001,a MMPs inhibitor,on BRB permeability.Methods Twenty-four adult clean SD rats were randomized to the control group,diabetic group and diabetes+GM6001 group according to randomized number table.Diabetic models were induced by the intraperitoneal injection of streptozotocin in the rats of the diabetic group and the diabetes + GM6001 group,and equal volume of citrate buffer was used in the same way in the rats of the control group.GM6001 10 μ1 (100 μ mol/L) was intravitreously injected in the third and fourteenth day after modeling in the diabetes+ GM6001 group,and equal volume of normal saline solution was injected in the same way in the control group and the diabetic group.The rats were sacrificed and eyeballs were extracted 1 month after injection,and the relative expressions of MMP-2 mRNA and MMP-9 mRNA in rat retinas were detected by reverse transcription PCR (RT-PCR).Evens blue (EB) was infused via the right jugular vein,and paraformaldehyde solution 1％ was then infused via left ventricle at the perfusion pressure 120 mmHg.The eyeballs were extracted 2 minutes later,and the leakage of EB in rat retinas was examined.Results RT-PCR electrophoresis exhibited the response bands of MMP-2 mRNA,and MMP-9 mRNA and GAPDH,with the gene size of 436,536 and 484 bp,respectively.The difference of the MMP2 mRNA and MMP-9 mRNA was statisticaly significant (F =20.336,P =0.000 ; F =8.742,P =0.002) ; and the relative expressions of MMP-2 mRNA and MMP-9 mRNA were significantly higher in the diabetic group and diabetes +GM6001 group than those in the control group (all at P＜0.01),and the relative expressions of MMP-2 mRNA and MMP-9 mRNA in the diabetes+GM6001 group was significantly reduced in comparison with the diabetic group(both P=0.01,P=0.02).The standardized EB content in the retinas of the control group,diabetic group and diabetes+ GM6001 group was (12.60±3.50) ng/mg,(26.52±7.14) ng/mg and (17.55±2.65) ng/mg,showing a significant difference (F=17.032,P＜0.01),and EB content in rat retinas in the diabetic group was higher than that in the control group (P=0.003),and that in the diabetes+GM6001 group was lower in comparison with the diabetic group (P=0.020).Conclusions Intravitreal injection of GM6001 can down-regulate the expression of MMP-2 mRNA and MMP-9 mRNA in diabetic rats and therefore protect BRB.