1.Promoting effect of connective tissue growth factor on expression of E-cadherin in human Tenon capsule fibroblasts
Chinese Journal of Experimental Ophthalmology 2015;33(8):695-698
Background Scarring of filtration channel following glaucoma filtering surgery is a main cause of the failure of the surgery.The proliferation,epithelial-mesenchymal transition and extracellular matrix remodeling of fibroblasts are thought to be the primary pathological mechanism of scarring.Connective tissue growth factor (CTGF) plays a promoting role in the formation of scar.Whether CTGF participates in mesenchymal-epithelial transition of human Tenon capsule fibroblasts (HTFs) is not clear yet.Objective This study attempted to investigate the effect of CTGF on the mesenchymal-epithelial transition of HTFs in vitro.Methods HTFs were cultured and passaged in high glucose DMEM medium with 10% fetal bovine serum,and the cells of generation 3-6 were used in this experiment.The cells were divided into the blank control group and CTGF-treated group and were routinely cultured in the blank control group.CTGF was added in the medium in the CTGF-treated group,with the final concentration 50 ng/ml CTGF.Immunofluorescence staining was used to identify and locate the expression of E-cadherin protein in the cells,and Western blot assay was employed to quantitatively analyze the expression level of E-cadherin protein in 48 hours after culture.Results The HTFs grew well with the spindle-like shape and vortex-like arrangement.The red fluorescence (E-cadherin protein) in the cytoplasm and blue fluorescence in the cellular nucleus were seen in the CTGF-treated group,but only nucleus with blue fluorescence were obtained in the blank control group.Western blot assay showed that the E-cadherin protein expression was absent in the blank control group,however,the relative expression level of E-cadherin protein in the cells was 0.63± 0.08.Conclusions E-cadherin protein is not expressed in fibroblast derived from mesenchymal tissue.However,CTGF can induce the expression of E-cadherin in HTFs.This study suggests that CTGF promotes the mesenchymal-epithelial transition of HTFs in vitro.
2.A GO analysis of gene expression on the inhibitory effect of endostatin on mouse choroidal neovascularization
Journal of Xi'an Jiaotong University(Medical Sciences) 2014;(3):333-337
Objective To study the gene expression on the inhibitory effect of endostatin on choroidal neovascularization in C57BL/6J mice and the mechanisms.Methods Photocoagulation using 532 nm diode laser was performed to establish a CNV model in mice’s eye.The mice were randomly divided into three groups:blank group (model group);endostatin group (experimental group),in which endostatin of 0.01 mg/2μL was given intravitreally;and saline group (control group),which received intravitreal injection of 2μL of 0.9 g/L saline.Four samples selected respectively from experimental group and control group were used to complete the gene expression profiling analysis.By comparing the differences of gene expression between the two groups,we selected the genes with expression difference ≥ 1.5 times and P ≤ 0.05. Results CD105 marking showed that choroidal neovascularization was significantly lower in endostatin group than in control group.The gene expression analysis showed that 1 1 6 genes were up-regulated and 1 0 6 genes were down-regulated in endostatin group compared with control group.GO analysis indicated that endostatin could inhibit cell activity and growth,but did not initiate the activity of the immune system,and even suppress it.Conclusion Endostatin can inhibit the activity and locomotion of endothelial cells and synergically inhibits the immune system,thus suppressing choroidal neovascular-ization.
3.Changes of cytokines in the aqueous humor following Avastin intravitreous injection
Jinan XIAO ; Qingyu BAI ; Anming XIE
Chinese Journal of Tissue Engineering Research 2014;(42):6805-6810
BACKGROUND:Studies on the inhibitory mechanism of Avastin for proliferative diabetic retinopathy are mostly confined to the vascular endothelial growth factor, while connective tissue growth factor and angiogenesis inhibitor also play an important role in the disease.
OBJECTIVE:To evaluate the changes of cytokines in the aqueous undergoing Avastin intravitreous injection or not prior to pars plana vitrectormy in the treatment of proliferative diabetic retinopathy.
METHODS:Thirty patients with proliferative diabetic retinopathy (30 eyes) were selected and randomly divided into groups A, B and C, each group containing 10 patients. Group A was treated with single pars plana vitrectormy;and group B and group C had 0.05 mL/1.25 mg and 0.1 mL/1.25 mg Avastin via intravitreous injection at 7 days before pars plana vitrectormy, respectively. Aqueous humor samples were col ected during pars plana vitrectormy. The levels of vascular endothelial growth factor, connective tissue growth factor and pigment epithelium-derived factor were assessed by using ELISA technique.
RESULTS AND CONCLUSION:The levels of connective tissue growth factor and pigment epithelium-derived factor in the aqueous humor of two Avastin injection groups were increased significantly compared to the group with single pars plana vitrectormy. However, the level of vascular endothelial growth factor was lower in the Avastin injection groups. In addition, there was no significant difference between two Avastin injection groups. These results reveal that intravitreous injection of Avastin assisted vitrectomy for proliferative diabetic retinopathy is more effective than single pars plana vitrectormy treatment. The underlying mechanisms may be that Avastin could inhibit the intraocular angiogenesis by not only reducing the secretion of some growth factors but also increasing the levels of anti-angiogenesis related cytokines.
4.Characteristics of CT imaging of adult ischemic moyamoya disease
Anming XIE ; Yaojun DING ; Gongjie LI
China Medical Equipment 2015;(10):12-15
Objective: To study the characteristics of CT imaging of adult ischemic type moyamoya disease, involving CT plain scan, CT perfusion imaging (CTP) and CT angiography (CTA). Methods:A retrospective analysis was made of the imaging data of 20 adult ischemic type moyamoya patients, including CT plain scan, CT perfusion imaging and CT angiography. CT vascular imaging features were graded I-VI with reference to Suzuki vascular grading. Results:Among the 20 patients with adult ischemic type moyamoya , ①CT plain scan: Old infarct lesions occurred in 13 cases(65%), 1 case suffered from acute cerebral infarction(5%), and negative patients totalled 6 (30%);②CT perfusion:5 cases(25%) were normal perfusion, 15 cases(75%) showed obviously low perfusion and local high perfusion;③CT angiography:patients of grade I to VI were respectively 1, 2, 5, 6, 4 and 2. Conclusion:Cerebral infarction lesions associated with adult ischemic type moyamoya disease are distributed mainly in the frontal and parietal cortex, or in watershed regions. Cerebral perfusion is characterized by normal or uneven blood perfusion, especially low perfusion. Vascular imaging manifests mostly degree III and IV, which belong to the middle phase of moyamoya disease.
5.Changes of cytokines in the aqueous humor following Avastin intravitreous injection in early diabetic retinopathy rats
Jinan XIAO ; Qingyu BAI ; Anming XIE
Chinese Journal of Tissue Engineering Research 2015;(18):2933-2939
BACKGROUND:Studies on the inhibitory mechanism of Avastin for early diabetic retinopathy rats are mostly confined to the vascular endothelial growth factor, while connective tissue growth factor and pigment epithelium-derived factor also play an important role in the disease.
OBJECTIVE:To evaluate the changed cytokines of aqueous humor and significance of Avastin intravitreous injection in early diabetic retinopathy rats.
METHODS:Rat models of early diabetic retinopathy were established by inducing with streptozotocin for 10 weeks. Avastin (1.25 mg and 2.5 mg) and physiological saline were injected in the vitreous space.
RESULTS AND CONCLUSION:Enzyme linked immunosorbent assay results revealed that compared with the physiological saline injection group, mass concentration of vascular endothelial growth factor was reduced, mass concentrations of pigment epithelium-derived factor and connective tissue growth factor was increased in aqueous humor of the two Avastin injection groups (P<0.05), but no significant difference in concentration of above cytokines was detected between the two groups (P>0.05). Results verified that Avastin intravitreous injection decreased vascular endothelial growth factor levels, decreased pigment epithelium-derived factor mass concentration and increased connective tissue growth factor levels in rat models of early diabetic retinopathy.
6.Collateral circulation characteristics of CT angiography imaging of adult ischemic type moyamoya disease
Anming XIE ; Yaojun DING ; Gongjie LI
China Medical Equipment 2016;13(4):64-66,67
Objective:To explore the characteristics of collateral circulation of moyamoya disease in CT angiography imaging.Methods: Data of 120 moyamoya disease patients diagnosed by the 94th hospital were collected. All the patients underwent CT angiography imaging and were divided into groups according to compensatory ways of collateral circulation. Lightspeed VCT was used in all patients to conduct CTA check, and assessed the clinical performance of 4 groups. Results: Group 1 consisted of 15 cases (12.5%) of ischemic type moyamoya disease. Group 2 had 53 cases (44.2%), 8 cases were ischemic type moyamoya disease, and 45 cases were beeding type group moyamoya disease. Group 3 had 38 cases (31.7%), who showed moyamoya vessel formation in the bottom of the brain, 7 of these cases were ischemic moyamoya disease, and 31 were bleeding type moyamoya disease. Group 4 had 14 cases (11.6%), all of whom belonged to bleeding type moyamoya disease, characterized by ophthalmic artery, temporal artery, middle meningeal artery, occipital artery communicating with terminal cortex intracranial vascular.Conclusion: Compensatory characteristics of collateral circulation vessels were closely correlated with the types of moyamoya disease.
7.Comparative analysis on the MR imaging characteristics between Ischemic moya-moya ;disease and hemorrhagic moya-moya disease
Anming XIE ; Yaojun DING ; Gongjie LI
China Medical Equipment 2015;(1):35-37,38
Objective: To improve the accuracy of forecasting hemorrhagic moya-moya disease by analyzing the difference in MR imaging between ischemic moya-moya disease and hemorrhagic moya-moya disease. Methods: Retrospective analysis was conducted of clinical and MR imaging data of 64 patients with moya-moya disease between 2009 and 2014 years in Hospital 94 of PLA. Results: Among the 64 patients aged 26 to 49 (average age was 38.2), 21 cases (32.8%) were diagnosed with ischemic moya-moya diseases, while 16 cases (76.2%) diagnosed with hemorrhagic moya-moya diseases, ischemic lesions were distributed mainly in frontal and parietal area, while hemorrhagic lesions were mainly distributed in the dorsal thalamus (28 cases, 65.1%), in the basal ganglia (9 cases, 20.9%), in the simple intraventricular (4cases, 9.3%) and in pure subarachnoid (2 cases, 4.6%). In the ischemic-typed moyamoya disease and hemorrhagic-typed moyamoya disease, cerebral bottom dorsal smoke abnormal vascular network, anterior choroidal artery and callosal artery thickening of the posterior cerebral artery, cortical pial vascular thickening, thickening of vascular branches of ophthalmic artery and external carotid artery thickening were respectively occurred in 15 cases of 28 branch (71.4%) and 38 cases of 62 branches (88.4%), 12 cases with 24 branches (57.1%) and 35 cases with 45 branches (81.4%), 8 cases with 16 branches (38.1%) and 30 cases with 58 branches (69.8%), 5 cases with 10 branches (23.8%) and 13 cases of the 24 branch (30.2%), 7 cases with 11 side branches (33.3%) and 27 patients with 54 branch (62.8%). Conclusion:The tortuous and dilated choroid artery and abnormal hyperplasia vascular network in skull base are the main causes of bleeding in moya-moya diseases.
8.Changes of cytokines in aqueous humor of patients with proliferative diabetic retinopathy after intravitreal bevacizumab and their correlation analysis
Guolong DING ; Anming XIE ; Jianqin LEI ; Guopeng DING ; Yaguang HU
Recent Advances in Ophthalmology 2017;37(4):358-361
Objective To study the changes of cytokines in aqueous humor of patients with proliferative diabetic retinopathy (PDR)after intravitreal bevacizumab (IVB) and the correlation of these cytokines.Methods Fifty-six patients(63 eyes) with severe PDR underwent pars piana vitrectomy (PPV) with IVB pretreatment in our hospital from April,2014 to February,2015 were collected.All the patients received IVB therapy from 3 days to 7 days before PPV.Aqueous humor samples were collected at the time of IVB and PPV treatment.The concentrations of vascular endothelial growth factor (VEGF),interleukin-6 (IL-6),interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1) were measured by cytoinetric bead array (CBA).The changes of these cyto-kines after IVB were analyzed by using Wilcoxon rank sum test for paired data.The correlation between these cytokines was analyzed by using Spearman rank test.Results From 3 days to 7 days after IVB,the concentration of VEGF in aqueous humor was (6.57 ± 12.29)pg · mL-1,lower than the pre-operation of (278.71 ±340.20)pg · mL-1,the difference was statistically significant (P < 0.05).The concentration of IL-8 was (385.60 ± 1099.47) pg · mL-1,higher than pre-operation of(298.87 ± 1005.79) pg ·mL-1,the difference was statistically significant (P < 0.05).IL-6 and MCP-1 were increased after IVB,but the difference was not statistically significant (all P > 0.05).VEGF,IL-6,IL-8 and MCP-1 had an obviously relationship between each two factors(all P < 0.05).Conclusion Angiogenesis in patients with PDR are regulated by a variety of cytokines,and targeted reduction of VEGF may compensatory increase the concentration of other inflammatory cytokines.
9.Correlative factors analysis of affecting anatomical reattachment and vision restoration after scleral buckling surgery
Lei ZHU ; Lin LI ; Xiaoyan TIAN ; Tao LIU ; Meng CHEN ; Pei XU ; Jing ZOU ; Anming XIE
Recent Advances in Ophthalmology 2017;37(2):167-171
Objeetive To analyze the effect of treatment of rhegmatogenous retinal detachment(RRD) by scleral bucking as well as the relative risk factors affecting the anatomical reattachment and visual recovery.Methods One hundred and fortyeight patients (148 eyes) with RRD treated by sclera buckling surgery in our hospital during January 2012 to January 2016 were retrospectively analyzed.The rate of postoperative retinal anatomic reattachment,the best corrected visual acuity (BCVA) and complications were observed.Logistic regression analysis was performed to analyze the correlative factors affecting the anatomical reattachment and postoperative vision restoration.R~ults Retinal reattachment achieved in 91.9% after initial surgery and the final success rate for anatomic reattachment was 97.3% assessed with ophthalmoscope and fundus photography.But these two rates were assessed with the optical coherence tomography (OCT) were 60.1% and 80.4% respectively.Single factor Logistic regression analysis showed that retinal detachment was affected by multiple breaks and Grade C1 PVR(all P <0.05);Single factor Logistic regression analysis showed that preoperative BCVA,course of disease,retinal detachment range,macular involvement or not had an impact on the postoperative recovery of BCVA (all P < 0.05),preoperative age,refractive status,releasing retinal fluid or not,intravitreal gas injection,combined scleral buckling,and postoperative subretinal fluid,all of these factors had no effect on BCVA recovery after surgery (all P > 0.05).And through multiple factors Logistic regression analysis,preoperative BCVA was an independent risk factor for BCVA recovery after surgery (P < 0.05).Conclusion Scleral bucking is an effective technique for managing RRD,but multiple breaks and Grade C1 PVR are significant risk factors for anatomic.Preoperative BCVA,course of disease,retinal detachment range,macular involvement or not have the impact on the BCVA recovery after scleral buckling,and the preoperative BCVA is the key factor.Early diagnosis and early treatment as well as protecting the preoperative visual acuity can improve prognosis.
10.Biological behavior changes and intergrin-linked kinase (ILK) expression after exposure to TGF-β2 in human Tenon fibroblasts
Lijun CUI ; Na SUN ; Yunlin TIAN ; Qun WANG ; Cheng PEI ; Anming XIE
Recent Advances in Ophthalmology 2017;37(7):605-609
Objective To investigate the biological behavior and expression of intergrin-linked kinase (ILK) after exposure to transforming growth factor beta 2 (TGF-β32) in human Tenon fibroblasts (HTFs).Methods The primary ceils of HTFs were cultured by tissue attached culture method and identified by immunofluorescence analysis with Vimentin and keratin.The proliferation levels of HTFs induced by different concentrations of TGF-β2 were analyzed by MTT.The α-smooth muscle actin (α-SMA) and ILK mRNA expression were analyzed by quantitative Real-time PCR.The protein expression of α-SMA,ILK and E-cadherin were analyzed by Western Blot,and the protein expression of α-SMA and E-cadherin were also analyzed by immunofluorescence staining.Results MTT analysis showed that the optical density levels of 5.0 μg · L-1 and 10.0 μg · L-1 TGF-β2 were significantly higher than those of 0.1 μg · L 1,1.0 μg · L-1 and the control after exposure for 48 hours and 72 hours,and all these optical density levels were significantly higher than that for 24 hours (all P <0.05).The expression of α-SMA and ILK mRNA increased significantly when cells were treated with 5.0 μg · L-1 TGF-β2 for 48 hours in comparison with the control group (all P < 0.05).The protein of α-SMA,ILK and E-cadherin were expressed both in TGF-β2 treated groups and control group,and TGF-β2 up-regulated the expression of them.There were significant differences when compared with the control group (all P < 0.05).Immunofluorescent staining showed that α-SMA and E-cadherin were detected in TGF-β2 treated groups.α-SMA expressed in cytoplasm,while E-cadherin both in cytoplasm and nucleus.Conclusion TGF-β2 can induce the proliferation,transdifferentiation and adhesion of HTFs,and up-regulate the expression of ILK in vitro,suggests that ILK may play a role in the process of scar formation after glaucoma filtration surgery.