Objective To observe the effect of intravitreal injection of bevacizumab (Avastin, IVB) on the expression of integrin-linked kinase (ILK) in fibrovascular membranes and the number of vascular endothelial cells (VECs) in proliferative diabetic retinopathy (PDR). Methods Twenty-four fibrovascular membrane samples were collected during pars plana vitrectomy in 24 patients with PDR. 12 PDR patients had received a single 1.25 mg IVB 7 days preoperatively (bevacizumab group), the other 12 patients (non-bevacizumab group) had not received IVB. For each of 24 fibrovascular membranes specimen, the number of VECs in the membranes were counted after staining with hematoxylin-eosin and von willebrand factor. Expressions of ILK in the fibrovascular membranes were detected through immunohistochemistry analysis. Results Immunohistochemistry revealed that ILK was highly expressed in all of 24 fibrovascular membranes of PDR.The average optical density of ILK expression level in bevacizumab and non-bevacizumab group were (127.78plusmn;15.08) and (129.03plusmn;16.26) respectively, the difference was not statistically significant (t=0.330，P=0.745).The number of VECs in fibrovascular membranes in bevacizumab and non-bevacizumab group were 21.50plusmn;3.94 and 41.33plusmn;7.44 respectively, the difference was statistically significant (t=3.872，P=0.003). Conclusions ILK was expressed in fibrovascular membranes of PDR. IVB can decrease the number of VECs during the process of PDR, but it can not affect the expression of ILK protein.
ObjectiveTo observe the efficacy and safety of individual dose of intravitreal conbercept (IVC) in the treatment of retinopathy of prematurity (ROP) before type 1 threshold.MethodsA retrospective case study. From January to July, 2019, 23 cases (46 eyes) of children with type 1 pre-threshold ROP were included in the study. Among them, 14 cases (28 eyes) were male and 9 cases (18 eyes) were female. The mean gestational age at birth was 28.06±1.73 weeks. The average birth weight was 1.14±0.19 kg. The mean corrected gestational age was 34.38±1.41 weeks at the time of first intravitreal injection of IVC. The axial length (AL) of children was measured by A-mode ultrasound before IVC for the first time. According to the calculation of AL, the corresponding injection dose range was 14.23-16.19, 16.20-17.57, 17.58-18.63 mm and the injection dose of IVC was 0.015, 0.020, 0.025 ml (including IVC was 0.15, 0.20, 0.25 mg, respectively). The first IVC dose was 0.015 ml. On the first day before IVC and on the first and seventh days after IVC, 2 ml of arterial blood was taken from children, serum vascular endothelial growth factor (VEGF) concentration was detected. The follow-up time after treatment was ≥1 year. After one year of follow-up, the effective rate and recurrence rate of IVC for the first time were tested by χ2 tests. The short-term changes of injection times, injection intervals, retinal vascularization time and serum VEGF concentration in children were tested by t test.ResultsRetinal neovascularization subsided and vascular buckling decreased in all eyes. Iris neovascularization subsided, 1-3 weeks after IVC for the first time. Within one year after the first IVC, 16 eyes underwent IVC twice with or without new blood vessels at the junction of the vascular area. The average corrected gestational age was 40.56±3.81 weeks. The injection dose of IVC was 0.015 ml and 0.020 ml for 2 eyes and 14 eyes, respectively.The mean interval from IVC for the first time was 40.89±8.99 days. Of the 16 eyes who underwent IVC twice, 8 eyes showed neovascularization again in the retinal area with or without blood vessels. The average corrected gestational age was 43.00±1.41 weeks. The injection dose of IVC was 0.020 ml and 0.025 ml for 3 eyes and 5 eyes, respectively. The mean interval of the second IVC was 28.60±6.07 days. The mean interval from the first IVC was 69.20±12.40 days. At the end of follow-up, all eyes were treated effectively (100%, 46/46). The mean time of retinal vascularization was 46.31±3.42 weeks. The average number of injections was 1.52±0.76. On the first day before IVC and on the first and seventh days after IVC, the average serum VEGF concentrations were 111.21±148.71, 25.60±27.71 and 42.99±38.01 pg/ml, respectively. Serum VEGF concentration was significantly lower than that before IVC on the 1st and 7th day after IVC (Z=−4.054, −2.779; P＜0.05). Serum VEGF concentration was higher 7 days after IVC than 1 day after IVC, and the difference was statistically significant (Z=−2.505, P＜0.05). All eyes were not treated by laser photocoagulation or vitrectomy. No eye complications such as lens opacification, endophthalmitis and retinal detachment related to drugs or treatment methods were found in all patients.ConclusionIntravitreal injection of individualized dose of IVC is effective in the treatment of type 1 pre-threshold ROP. Seven days after treatment, serum VEGF concentration of patients’serum decreases.
Objective To observe the expression of matrix metalloproteinase（MMP-2, MMP-9 and vascular endothelial growth factor (VEGF) in retinoblastoma (RB) and its relationship with the differentiation and optic nerve infiltration of RB.Methods Forty paraffin specimens of pathological confirmed RB were studied. They were divided into differentiated group (15 cases) and undifferentiated group (25 cases) , optic nerve infiltration group（13 cases） and without optic nerve infiltration group（27 cases）. The expression of MMP-2, MMP-9 and VEGF were detected by immunohistochemistry, their relationships with the differentiation and optic nerve infiltration were also analyzed.Results The positive rate of MMP-2, MMP-9 and VEGF expression in 40 RB cases were 52.5％，57.5％ and 72.5% respectively．The expression of MMP-2, MMP-9 and VEGF in the undifferentiated group were significantly higher than those in the differentiated group (chi;2=9.037, 9.253, 8.095; P＜0.05). The expression of MMP-2, MMP-9 and VEGF in RB with optic nerve infiltration group were significantly higher than those in RB without optic nerve infiltration group (chi;2=11.045,10.243, 8.956；P＜0.05). The expression of MMP-2，MMP-9 had a positive correlation with the expression of VEGF in RB (r=0.126,0.314；P＜0.05). Conclusions MMP-2, MMP-9 and VEGF expressed in RB tumor tissues. The expression of MMP-2, MMP-9 has a positive correlation with the expression of VEGF. The levels of MMP-2, MMP-9 and VEGF expression are related to optic nerve infiltration of RB cells.
Objective：To investigate the role of 17beta; estradiol on th e expressi on of vascular endothelial growth factor (VEGF) and on the releasing rate of lac tate dehydrogenase (LDH) in cultured anoxiainjured human retinal pigment epit h eliual (RPE) cells. Methods:Established the anoxiainjuried m odel of human RPE c ells with Cobalt Chloride (CoCl2) after RPE cells were pretreated with 17beta;E 2 and tamoxife, 17beta;E2 antagonist. The expression of VEGF mRNA was detecte d by re v erse transcriptionpolymerase chain reaction technique (RTPCR). The cultured RP E cells were divided into four groups: normal control group, anoxiainjured gro u p, 17beta;E2 pretreatment group and 17beta;E2 with tamoxifen pretreatment grou p. The releasing rate of LDH was detected by chromatometry. The expression of VEGF pro tein were detected by cellular immunohistochemistry. Results:T he expression of VEGF and LDH releasing rate were higher in anoxiainjured grou p than that in nor m al control group (P＜0.05), and were lower in 17beta;E2 pretreatment group than th at in anoxiainjured group （P＜0.05）. When the effect of 17beta;E2 was o bstructe d by tamoxifen, the expression of VEGF and LDH releasing rate increased but didn prime;t differ much from which in anoxiainjured group (P＞0.05). Conc lusion:The ex pression of VEGF increases in anoxiainjured human RPE cells. 17beta;E2 can do wnr egulate the expression of VEGF and decrease the releasing rate of LDH, which can be blocked by tamoxifen.
Objective To investigate the influence of vascular endothelial growth factor (VEGF) antagonist bevacizumab on the growth of human choroidal melanoma (CM) OCM-1 cell xenografts in nude mice, and to explore the probable mechanism.Methods OCM-1 cells were subcutaneously implanted on 18 nude mice to establish ectopic model of human CM. The nude mice with the tumor of 5 mm in diameter were randomly divided into three groups: untreated group (group A), normal saline （NS） group (group B), drug treated group (group C). Bevacizumab was intraperitoneally injected for 14 consecutive days in group C, and the same volume of NS was used at a same way in group B. The volume and weight of implanted tumor as well as inhibitory rates of drug on tumor were calculated, ki67 and survivin proteins were measured with immunohistochemistry, and the mRNA expression of VEGF and survivin were assessed by RT-PCR.Results The volume and weight of tumor was (598.86plusmn;321.81) mm3, (0.66plusmn;0.15) g; (1 715.15plusmn;278.16) mm3, (1.54plusmn;0.39) g and (1 750.23plusmn;206.36) mm3, (1.54plusmn;0.31) g in groups C, A and B, respectively. There were significant differences between group C and A (F=34.53, P=0.00) and group C and group B (F=8.69, P=0.01). The inhibitory rate of these three groups were 57.14%, 5.31%, 6.25%, respectively, and the proliferation index (PI) of ki67 in these three groups were (51.85plusmn;1.32)%, (46.30plusmn;1.39)%, (27.90plusmn;0.90)%, respectively, there were significant differences in ki67 PI between C group and A or B group (H=15.17, P=0.00). The expression of survivin mRNA was (0.49plusmn;0.02), (0.82plusmn;0.05) and (0.61plusmn;0.05) in groupss C, A and B, respectively, there were significant differences between C group and A or B group (F=15.17, P＜0.05) . The expression of VEGF mRNA was (0.32plusmn;0.08), (0.73plusmn;0.07), (0.80plusmn;0.04) in groups C, A and B, significant difference was found between group C and A or B group (F=12.05，P＜0.05). Conclusion Bevacizumab can inhibit the growth of human CM in nude mice probably by inhibiting the activity of VEGF and downregulating survivin expression of the tumor as well as inhibiting the growth of the tumor.
Anti-vascular dndothelial growth factor (VEGF) drugs have open up a new treatment channel for ocular neovascular diseases. A lots of clinical data has proved that anti-VEGF drugs are effective and safe. But we should also notice that long-term and excessive usage of anti-VEGF drugs brings some new problems and complications, and even affect the normal ocular physiological process of the angiogenesis and retinal blood flow. So, it is necessary to pay attention to the problems and potential risks of excessive usage of anti-VEGF therapies for ocular neovascular disease.
Objective To observe the influence of rAAV-mediated antisense vascular endothelial growth factor (rAAV-aVEGF165) on the expression of retinal VEGF in diabetic rats. Methods 40 Sprague-Dawley rats induced diabetic rat model by intraperitoneal injection with streptozotocin (STZ). 32 rats were involved in study besides death and blood sugar recovery in experimental process, 16 spragud-Dawleg (SD) rats were received intravitreal injection with rAAV-aVEGF165 (1010 pfu) as experimental group, another group of Sprague-Dawleg (SD) rats were injected with phosphate buffered saline (PBS) as control group. One and five month after model establishment, the expression of retinal VEGF was evaluate by immunhistochemistry and Western blot; the retinal vasular was examined by transmission electron microscopy. Results On 1 month,the expression of retinal VEGF was lowest in each group. On 5 month, the expression of retinal VEGF was decreased in experimental group which compared to control, the difference are statistically significant （t=23.87,Plt;0.01）. The transmission electron microscopy results showed that retina has no obvious chages in experimental group, however,contral group showed fragmental thickening and splitting of basement membrane, swelling and deformation of endothelia cells,fingerlike prcess into the capillary cavity,and uneven distibution of heterochromatin in pericytes. Conclusion rAAV-aVEGF165 can reduce the expression of retinal VEGF thereby preventing occurrence and development of diabetic retinopathy. rAAV is an effective vectors of eye antisense gene. （Chin J Ocul Fundus Dis，2008，24：255-258）
ObjectiveTo study the changes and correlation of cytokines in aqueous humor before and after intravitreal injection of conbercept (IVC) treatment in patients with proliferative diabetic retinopathy (PDR).MethodsA prospective clinical study. From March to December 2019, 36 patients (42 eyes) of PDR patients treated with IVC combined with pars plana vitrectomy (PPV) (the observation group) and 27 patients (31 eyes) underwent cataract surgery in the same period (control group) in Department of Ophthalmology of the First Affiliated Hospital of Guangzhou University of Chinese Medicine were included in this study. Before PPV 5-7 days, IVC treatment was performed, and the aqueous humor were extracted during IVC and second-stage PPV in the observation group. The aqueous humor was extracted during cataract surgery in the control group. Luminex assay was used to detect VEGF-A, placental growth factor (PLGF), platelet-derived growth factor-AA (PDGF-AA), platelet-derived growth factor-BB (PDGF-BB), angiopoietin-like protein 4 (ANGPTL4), IL-6, IL-8, IL-1β, monocyte chemoattractant protein-1 (MCP-1), intercellular adhesion molecule-1 (ICAM-1), tumor necrosis factor-α (TNF-α) cytokine expression. For normally distributed data, the independent sample t test was used for comparison between two independent samples; for non-normally distributed data, the Wilcoxon rank sum test was used for comparison between two independent samples. The correlation analysis used Spearman rank correlation test.ResultsBefore IVC treatment, the concentrations of VEGF-A, PLGF, PDGF-AA, ANGPTL4, IL-6, IL-8, MCP-1 and ICAM-1 in the aqueous humor of PDR patients were significantly higher than those in the control group (P＜0.05). After IVC treatment, the concentration of VEGF-A in the aqueous humor was significantly lower than that before treatment, and the concentrations of ANGPTL4 and IL-8 were significantly higher than those before treatment (P＜0.05). There were no significant differences in the concentrations of PLGF, PDGF-AA, PDGF-BB, IL-6, IL-1β, MCP-1, ICAM-1 and TNF-α before and after IVC treatment (P＞0.05). Before IVC treatment, the concentration of VEGF-A was positively correlated with PLGF, PDGF-AA, PDGF-BB, ANGPTL4, IL-6, IL-8, MCP-1 and TNF-α (P＜0.05).ConclusionsIVC treatment can reduce the concentration of VEGF-A and increase the concentrations of ANGPTL4 and IL-8 in aqueous humor in PDR patients before PPV.
ObjectiveTo analyze the concentrations of vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF) in aqueous humor of patients with proliferative diabetic retinopathy (PDR) before and after intravitreal injection of ranibizumab. MethodsTwenty-five eyes of 20 PDR patients were collected as the PDR group. Twenty-five eyes of 21 senile cataract patients were collected as the control group. There were no statistical significance in gender (χ2=0.223), age (Z=-1.555) and intraocular pressure (Z=-0.225) between the two groups (P > 0.05). Samples of aqueous humor (0.1 ml) were collected just before and 7 days after the injection of ranibizumab in PDR group. Samples of aqueous (0.1 ml) humor were collected just before cataract surgery in control group. The concentrations of VEGF and PEDF in the aqueous humor were measured by enzyme-linked immunosorbent assay. ResultsThe VEGF and PEDF concentration in the aqueous humor were reduced significantly after intravitreal injection of ranibizumab in PDR group (Z=-4.072, -4.319; P < 0.05). The concentrations of VEGF and PEDF in the aqueous humor before intravitreal injection of ranibizumab in PDR group were significantly higher than the control group (Z=-5.228, 4.706; P < 0.05). The VEGF concentration in the aqueous humor after intravitreal injection of ranibizumab in PDR group were similar to control group (Z=-1.557, P > 0.05). However, the concentration of PEDF in the aqueous humor after intravitreal injection of ranibizumab in PDR group still higher than control group (Z=-2.475, P < 0.05). The ratio of VEGF/PEDF before and after intravitreal injection of ranibizumab was statistically different (Z=-2.058, P < 0.05), but was the same between PDR group and control group (Z=-0.456, -0.844; P > 0.05). The aqueous humor concentrations of VEGF and PEDF were not significantly correlated with each other, neither in PDR group (r=-0.195, -0.174; P > 0.05) nor in control group (r=-0.286, P > 0.05). ConclusionsAqueous humor concentrations of VEGF and PEDF are significantly elevated in eyes with PDR. Intravitreal injection of ranibizumab significantly decreased the VEGF and PEDF in the aqueous humor after 7 days.
Proliferative vitreoretinopathy (PVR) is a common complication and major cause of blindness of ocular trauma. Many cytokines, including vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF), participate in the process of the pathogenesis of traumatic PVR. VEGF competitively inhibits binding of PDGF to its receptor (PDGFRα), enables indirect activation of PDGFRα by non-PDGF ligands, resulting in reduced p53 expression, cell proliferation and migration, which is a key point in the pathogenesis of traumatic PVR.