Objective To observe the therapeutic effect of ultrasonic microbubble combined with bevacizumab (Avastin) on choroidal neovascularization induced by photocoagulation in rabbits.Methods CNV was induced by photocoagulation with argon laser in 30 rabbits (60 eyes).All of the rabbits underwent fundus fluorecein angiography (FFA) 21 days after photocoagulation; 6-8 hours later, 3 rabbits were randomly chosen to be executed to having the immunohistochemical examination.Twenty one days after photocoagulation, 27 rabbits were divided randomly into 3 groups: bevacizumb, ultrasonic microbubble + bevacizumb,and control group; each group has 9 rabbits (18 eyes).The rabbits in control group had no interference treatment; while the rats in bevacizumb and ultrasonic microbubble + bevacizumb group underwent injection with bevacizumb or ultrasonic microbubble + bevacizumb respectively.FFA was performed on all of the rabbits 7,14,and 28 days after photocoagulation to observe the inhibition of CNV; immunofluorecence and Western blot were used to detect the expression of VEGF in retina and choroid.Twentyeight days is the time point to determine the therapeutic efficacy. The expression of VEGF and the results of FFA were the sdandards of the judgement of therapeutic efficacy.Results Proliferaion of CNV to the retinal inner layer and the obvious leakage of fluoresein in the photocoagulation area indicated that the model of CNV was set up successfully. Twenty eight days after injection,obvious fluorescent leakage was found in the control group, and the average fluorescent leakage in bevacizumab group differed much from the control group（t=16.2952，Plt;0.05）; while the difference between ultrasonic microbubble + bevacizumb group and bevacizumab group was also significant (t=4.7955，Plt;0.05) . At the same time point, the expression of VEGF in bevacizumab group detected by immunofluorecent assay and Western blot differed much from the control group (t=7.0327，9.2596；Plt;0.05),and the difference of VEGF between ultrasonic microbubble + bevacizumb group and bevacizumab group was significant（t=2.9724,17.1937；Plt;0.05）. this experiment show that ultrasound combined bevacizumab intravitreal injection of the therapeutic effect of CNV superior to other groups(Plt;0.01).Conclusion Ultrasound microbubble combined with bevacizumab injection may improve the therapeutic effect on CNV by inhibiting the expression of VEGF.
Objective To investigate auto-cortex of crystalline lens-induced neovascular epiretinal membrane（NVERM）by micro-injuring posterior c apsule of crystalline lens. Methods twenty four C57BL/6 mouse between 4-6 weeks were selected, and divided into two groups randomly: auto-cortex of crystalline group and the control group. The auto-cortex of crystalline group was treated by penetrating the posterior capsule of lens and washing out the lens cortex into the mouse vitreous using PBS (phosphate buffered solution), while the control group were injected PBS into vitreous merely. Clinical change s were followed by slit-lamp examination and photograph. The eye balls were enu cleated at the day of 3, 7, 14 and 28 after operation. Both HE and immunohistoch emistry were used to detect the pathological changes. Results postoperative one to three days, 11 of 12 mouse in autocortex of crystalline g roup, lens appear to alba turbid at different levels one after another, and then develop into highdensity chinaware white. Postoperative (po) three days, HE s taining shows cortex of lens debris transmigrated in vitreous cavity, and some o f which approached to internal limiting membrane and lead it to rough and discon tinue; Po7-14 days, the capillary in retina expanded, migrated and broke though t internal limiting membrane which got to the pro retina and became the new ves sels. And typical NVERM were observed. Po28 days, some vascularslike structure formed in vitreous cavity. None of mouse in control group developed NVERM. Conclusion Auto-cortex of crystalline lens can induced neovascular epiretinal membrane in C57BL/6 mouse. (Chin J Ocul Fundus Dis,2008,24:118-121)
ObjectiveTo analyze the regulative rule of mRNA of vascular endothelial growth factor (VEGF) in mice with oxygen-induced retinopathy, and to elucidate the possible mechanism of occurrence of neovascularization in retinopathy of prematurity (ROP).MethodsSixty 7-day-old C57BL/6J mice were divided into oxygen-induced retinopathy group and control group. In oxygen-induced retinopathy group, 36 mice were exposed to 75% oxygen for 5 days and then to room air for 5 days; in control group, 24 mice were raised in room air. Vascular perfusion of fluorescein and retinal stretched preparation were used to observe the morphologic changes of retinal vessels. Reversal transcriptionpolymerase chain reaction (RT-PCR) was used to observe changes of VEGF mRNA in each group. ResultsIn oxygen-induced retinopathy group, the morphologic characteristics of retinal vessels were the unperfused area at the center of superficial and deepseated vessels, and the neovascularization appeared at mid-peripheral retina after 2 days in relative hypoxia condition. The results of RT-PCR showed space-time corresponding relation between expression of VEGF and neovascularization, which meant that the transcription of VEGF mRNA decreased in hyperxia conditionand increased in relative hypoxia condition. ConclusionHypoxia is the main reason of occurrence of retinal neovascularization; increased expression of VEGF caused by relative hypoxia after hyperxia might be effective in reducing the occurrence of neovascularization in ROP.(Chin J Ocul Fundus Dis, 2005,21:292-295)
The therapeutic effects of panretinal cryotherapy(PRC)on proliferative diabetic retinopathy(PDR)were prospectively investigated in 80 eyes with PDR of 44 patients.Forty eyes with PDR(20 of them stage Ⅳ and 20 stage Ⅴ)received PRC operation.Of the 80 eyes,the other 40 ones with stage Ⅳ and Ⅴ similar to the formers were observed conservatively as controls.The follow up duration was 2 years.We found that in the cases of stage Ⅳ,no more remarkable visual loss was found after operation.There was a significant difference comparing with the control（P＜0.002）,and the retinal neovascularization regressed more noticeably than the control group（P＜0.001）.In the cases of stage Ⅴ,the incidence of the traction retinal detachment was 55% in the operated group,and was 20% in the control group.There was a statistic difference between them（P＜0.05）.The clearance of the vitreous hemorrhage was more rapid in the operated group than the control（P＜0.025）.The above results suggest that cryotherapy is suitable for the cases of earlier stage which cannot be performed with photocoagulation for any reasons,but not for the patients with advanced retinal proliferation.Photocoagulation for any reasons,but not for the patients with advanced retinal proliferation. （Chin J Ocul Fundus Dis,1993,9:148-151）
Objective To determine the expression of the growth factors and the receptors related to angiogenesis in the intraocular tissues incarcerating in the sclerotomy sites. Methods Ten specimens from prolapsing intraocular tissues in sclerotomy sites during vitrectomy were obtained and serially sectioned in cryostate and were stained with a group of polyclonal antibodies against vascular endothelial growth factor(VEGF), basic fibroblast growth factor (bFGF), platelet-derived growth factor-A(PDGF-A) and transforming growth factor-β1(TGF-β1) as well as their receptors by using a streptavidin peroxidase system. Results The tissues prolapsed from the sclerotomy sites were identified as retina(3 cases), vitreous tissues(3 cases), degenerated red blood cell components(2 cases), ciliary body(one case) and fibrous tissue(one case). All specimens expressed VEGF and bFGF as well as their receptors. PDGF-A, TGF-β1 and their receptors expressed in the most of specimens. The positive cells included retinal cells, ciliary non-pigmented epithelial cells and pigmented epithelial cells, fibrous cells and the cells in vitreous. Conclusions The intraocular tissues incarcerated in the sclerotomy entries express the growth factors and receptors related to angiogenesis. This might be one of the potential factors of developing anterior proliferative vitreoretinopathy. (Chin J Ocul Fundus Dis, 2002, 18: 34-37)
Purpose To investigate the status of proliferation and activation of vascular endothelial cells in preretinal neovascular membranes from patients with insulin dependent diabetetes mellitus(IDDM)by means of immunohistochemical techniques. Methods Status of vascular endothelial cells in 18 preretinal neovascular membranes from 18 patients with IDDM was studied by double-immunofluorescence technique and the alkaline phosphataes-anti-alkaline phosphatase(APAAP)technique and compared the findings with the main clinical features of the patients. Results Of 18 vascularized membranes,16(88.9%)contained proliferating endothelial cells (positive for proliferating vascular endothelial cell marker EN 7/44) and 14 (77.8%) were positive for endothelial cell activation marker anti-VCAM-1;furthermore,by using a double staining technique,we found that in 14 of the 16 cases(87.5%) the proliferating vascular endothelial cells were activated (expressing VCAM-1). Conclusion The proliferation and activation of the vascular endothelial cells of the newly formed vessels in preretinal neovascular membranes suggests the significance of the vascular endothelial cells in the pathophysiology and the progress of proliferative diabetic retinopathy. (Chin J Ocul Fundus Dis,1998,14:141-143)
Objective To compare two kinds of myofascial flap encapsulating adi pose-derived stromal cells (ADSCs) in adi pogenic efficacy in vivo, and to provide experimental basis for the efficient transplantation of free adi pose tissue. Methods ADSCs were isolated from the subcutaneous adipose tissue in the neck of 10 New Zealand rabbits (aged 3-4 months old, male and female, weighing 2.0-2.5 kg), and primary culture and subculture of ADSCs were conducted. When the cells at passage 3 covered 70%-80% of the bottom of the culture flask, BrdU (10 μg/mL) was appl ied to label the cells for 48 hours before performing immunofluorescence staining. Oil red O staining observation was conducted to thosecells 2 weeks after being induced towards adi pocyte, al izarin red staining observation was performed 3 weeks after being induced towards osteoblast, and alcian blue staining was conducted 2 weeks after being induced towards chondrocyte. Besides, after being induced towards adipocyte for 2 weeks, 1 × 107 ADSCs/piece at passage 3 labeled by BrdU was seeded into Col I (10 mm × 10 mm × 5 mm/piece) to prepare cell carrier complex. The experiment was divided into two groups: group A in which vascular pedicled dextral latissimus dorsi fascial flap was adopted to encapsulate the complex; group B in which dextral gluteus maximus fascial flap with no specific vessel pedicle was appl ied to encapsulate the complex. Rabbits in each group went through autogenous ADSCs transplant and self control. The implants were dislodged 8 weeks after operation, HE staining and immunohistochemistry staining were performed to testify cambium, the wet weight and micro vessel count of the cambium in each group were tested, immunofluorescence staining was performed to determine the origin of cambium and microvascular endothel ium. Results The nucleus of ADSCs positive for BrdU label ing showed green fluorescence under fluorescence microscope, with the positive label ing ratio of ADSCs above 90%. For ADSCs at passage 3, the formation of red l ipid droplets within cells was observed 2 weeks after being induced towards adipocyte, red calcium nodules were evident 3 weeks after being induced towards osteoblast, and highly congregated cell mass positive for alcian blue staining appeared 2 weeks after being induced towards chondrocyte. Eight weeks after operation, neogenetic blood vessel grew into scaffolds and no obvious fibreencapsulation was observed in group A, while few blood vessel grew into scaffolds in group B. The wet weight of cambium in group A and B was (0.149 5 ± 0.017 3) g and (0.095 3 ± 0.012 7) g, respectively, indicating there was a significant difference between two groups (P lt; 0.01). HE staining showed the formation of neogenetic adipose tissue and the growth of micrangium in the implant, and the degradation and absorption of scaffold. The micro vessel count of group A and B was 31.2 ± 4.5 and 19.3 ± 2.6, respectively, indicating there was a significant difference between two groups (P lt; 0.01). Eight weeks after operation, the immunofluorescence staining of cambium showed that the cell nucleus of regenerated adi pocytes and partial capillary endothel ium in groups A and B presented green fluorescence. Conclusion ADSCs encapsulated by vascular pedicled latissimus dorsi fascial flap and collagen protein scaffold complex has a higher adi pogenic efficacy in vivo than the gluteus maximus fascial flap with no specific vessel pedicle.
The effect and opportunity of argon laser photocoagulation for the retinal neovascularization in branch retinal vein occlusion in 30 patients were investigated with a control group of 34 patients received nonlaser but routine treatment. The results of the therapeutic effect demonstrated that the neovascularization disappeared completely in 23 cases and became smaller in 7 cases after laser photocoagulation. The incidnce of vitreous hemorrhage in laser group was 43.3% before laser treatment and none after treatment in the duration of observation,and 70.6% in control group. The progression of visual acuity after treatment in laser group was much better than in control group(P＜0. 005)at the time of the latest examination. We found the therapeutic effect was relation to the area, location of the neovascularization in retina,as well as whether the new vessels protruding into vitreous or not. （Chin J Ocul Fundus Dis,1994,10:195-198）
ObjectiveTo observe the efficacy of intravitreal injection of conbercept (IVC) in the treatment of type 1 macular neovascularization (MNV) with different types of pigment epithelial detachment (PED) in neovascular age-related macular degeneration (nAMD). MethodsA retrospective clinical study. From June 2018 to June 2021, 42 patients with 42 eyes of nAMD type 1 MNV patients with different types of PED diagnosed in the ophthalmological examination of the Department of Ophthalmology, General Hospital of Central Theater Command were included in the study. All eyes underwent best corrected visual acuity (BCVA) and optical coherence tomography (OCT). The OCT examination was performed with a 3D-OCT 2000 instrument from Topcon Company in Japan. The fovea was scanned, and the PED height (PEDH), PED area (PEDA), PED volume (PEDV), and central foveal thickness (CFT) were measured. According to the OCT image features of PED, the affected eyes were divided into serous PED (sPED), fibrovascular PED (fPED), and hemorrhagic PED (hPED), and were grouped accordingly. Among the 42 eyes, 16 (38.1%, 16/42), 14 (33.3%, 14/42), and 12 (28.6%, 12/42) eyes were in the sPED group, fPED group, and hPED group, respectively. All patients received IVC treatment once a month for 3 consecutive months, and then on-demand treatment after assessment. BCVA and OCT were re-examined 3, 6, and 12 months after treatment, and the changes of BCVA, PEDH, PEDA, PEDV, and CFT in the affected eyes before and after treatment were compared, and repeated measures analysis of variance was used for statistical analysis. ResultsAt 12 months after treatment, the PEDH, PEDA and PEDV of the affected eyes in the sPED group, fPED group and hPED group were significantly lower than those before treatment, and the difference was statistically significant (P<0.05). The difference in the degree of improvement was －318.67±258.09 μm, －6.50±6.33 μm2, －1.95±1.78 μm3 in the hPED group; －119.31±224.13 μm, －0.86 ±5.00 μm2, －0.56±1.64 μm3 in the sPED group; fPED group were －53.93±92.51 μm, －0.76±2.54 μm2, －0.19±0.46 μm3. The improvement degree of the affected eyes in hPED group was significantly greater than that in sPED group and fPED group, and the difference was statistically significant (F=5.918, 6.029, 5.494; P＜0.05). Compared with the BCVA and CFT before treatment, 12 months after treatment, the difference was statistically significant in the fPED group and the hPED group (P＜0.05); there was no significant improvement in the sPED group (P＞0.05). There was no significant difference in the BCVA of the affected eyes in the three groups compared with those before treatment (F=0.817, 0.741, 0.848; P＞0.05). ConclusionConbercept can effectively improve or stabilize the visual function and anatomical morphology of eyes with type 1 MNV in nAMD with sPED, fPED and hPED, among which the anatomical effect is better for hPED.
ObjectiveTo assess the occurrence of CNV in patients presenting with flat irregular pigment epithelial detachments (FIPED). MethodsForty-five patients (49 eyes) with FIPED on OCT were enrolled in this retrospective study. There were 25 males (28 eyes) and 20 females (21 eyes). The mean age was 61.022±9.292 years. FFA, ICGA, spectral domain OCT and OCT angiography (OCTA) were performed in all patients during the same period. The FIPED was defined as an irregular elevation of the RPE allowing distinct visualization of Bruch’s membrane on OCT B-scan. The abnormal vascular signals from the deep retinal layer to the choroid layer on OCTA was defined as CNV. The CNV was classified into a type 1 CNV and a type 2 CNV according to the OCT characteristics. The CNV was classified into a typical and occult CNV according to the characteristics of the FFA image. Of all 49 eyes, fundus angiography revealed 18 eyes (36.7%) with CNV, and 31 eyes (63.3%) with no characteristic signs of CNV. FFA examination found that CNV in 8 eyes (classic CNV in 1 eyes, occult CNV in 7 eyes), which confirmed by OCT were type 1 CNV; transmitted fluorescence in 41 eyes. ICGA examination showed that CNV-like hyperfluorescence spots in 18 eyes, suspicious hyperfluorescence spots in late stage in 20 eyes, and choroidal high permeability in 11 eyes, respectively; and 18 CNV eyes were confirmed to be type 1 CNV by OCT. To compare the detection of CNV by OCTA and fundus angiography. ResultsOf the 49 eyes with FIPED, OCTA detected 36 eyes (73.5%) of type 1 CNV, and full or partial strong reflex signals were seen in FIPED; 13 eyes (26.5%) were not associated with CNV, and some strong reflection signals were found in FIPED in 9 eyes, 4 eyes with weak reflection signal. The FFA was examined for 1, 7 eyes of the classic and occult CNV, which confirmed to be type 1 CNV by OCTA. Among the 18 eyes with CNV which detected by ICGA, OCTA also found type 1 CNV. Among the 20 eyes with ICGA’s late suspicious strong fluorescent spots, OCTA showed 17 eyes of type 1 CNV; in 11 eyes with high choroidal permeability, OCTA showed type 1 CNV in 1 eye. Among the 36 eyes with CNV which detected by OCT, there were SRD in 32 eyes, no SRD in 2 eyes and retinal interlamellar cavities in 2 eyes. ConclusionOCTA can detect 73.5% of FIPED eyes with CNV. Compared with traditional fundus angiography, OCTA has a higher detection rate of CNV under FIPED. The FIPED of the internal strong reflection signal has a certain diagnostic value for the type 1 CNV.