Objective To observe the ocular hemodynamic changes and flow direction of ophthalmic artery of patients with severe internal carotid stenosis (ICAS) and investigate the relationship between flow direction of ophthalmic artery and degree of stenosis. Methods Forty eyes of 40 patients with unilateral highgrade ICAS (29 eyes, 72.5%) and internal carotid artery occlusion (11 eyes, 27.5%) diagnosed by color Doppler flow imaging (CDFI) were enrolled in this study. There were 14 eyes (35.0%) with obvious ocular signs of ischemia, 26 eyes (65.0%) without obvious signs of ocular ischemia. The peak systolic velocity (PSV) of central retinal artery (CRA) was measured. The flow direction of the ophthalmic artery was observed by digital subtraction angiography (DSA). The PSV of CRA in eyes with different flow directions in the ophthalmic artery was comparatively analyzed. The relationship between flow direction of the ophthalmic artery and degree of stenosis was also analyzed. Results The PSV of CRA in ICAS eyes was (6.59plusmn;1.49) cm/s, which was decreased compared to fellow eye (8.95plusmn;1.35) cm/s, the difference was statistically significant (t=-7.24,P＜0.01). The PSV of CRA in eyes with signs of obvious ocular ischemia was (5.84plusmn;1.42) cm/s, which was decreased compared to eyes without signs of obvious ocular ischemia (7.00plusmn;1.39) cm/s, the difference was statistically significant (t=-2.49,P＜0.05). There were 15 eyes (37.5%) with retrograde flow in the ophthalmic artery, 25 eyes (62.5%) with forward flow of ophthalmic artery. The PSV of CRA in eyes with retrograde flow and forward flow of ophthalmic artery were (6.96plusmn;2.09), (7.01plusmn;1.42) cm/s, the difference was not statistically significant (t=-0.09，P＞0.05). Among 15 eyes with retrograde flow of ophthalmic artery, there were five eyes (33.3%) with unilateral high-grade ICAS, 10 eyes (66.7%) with internal carotid artery occlusion. The incidence rate of retrograde flow in the ophthalmic artery in eyes with internal carotid artery occlusion was higher than that in eyes with unilateral high-grade ICAS (P＜0.01). Conclusions The PSV of CRA in eyes with severe ICAS decreased compared to fellow eyes. The PSV of CRA in eyes with signs of obvious ocular ischemia also decreased compared to eyes without obvious signs of ocular ischemia. With the increase of the degree of the internal carotid artery stenosis, the incidence of retrograde flow of ophthalmic artery increased.
Objective To observe the serum lipid level of patients with branch retinal vein occlusion (BRVO). Methods A total of 71 BRVO patients (BRVO group) were enrolled in this study. The patients included 31 males and 40 females, with an average age of (52.75plusmn;10.2) years. All the patients were examined for visual acuity, slit lamp ophthalmoscopy combine with preset lens, fundus color photography and fundus fluorescein angiography (FFA) examination. Seventy-two age and sex matched normal subjects were enrolled in this study as control group. The subjects included 32 males and 40 females, with an average age of (53.10plusmn;9.5) years. The BRVO and control group were divided into four subgroup which including age with ＜40 years, 40-49 years, 50-59 years and ge;60 years. The plasma cholesterol and triglyceride level of BRVO group, control group, and age subgroups of BRVO and control group were comparatively analyzed. Results The average plasma cholesterol levels were (4.529plusmn;0.100) and (4.274plusmn;0.106) mmol/L in BRVO and control group, respectively. There was no difference between two groups (t=-1.738，P＞0.05). The average triglyceride levels were (1.500plusmn;0.129) and (1.319plusmn;0.095) mmol/L in BRVO and control group, respectively. There was no difference between two groups (t=-1.135，P＞0.05). There was no difference of average plasma cholesterol (t=-1.755, 1.850, -1.892, -0.507) and triglyceride (t=0.846, -0.074, -1.288, -1.887) level in age subgroups of BRVO and control subgroup (P＞0.05). Conclusion There is no significant difference of serum lipid level between BRVO patients and controls.
Objective To investigate the role of adenosine A2A receptor plays in retinal pathological neovascularization in mice. Methods A total of 202 mice were divided into room-air group (n=66) and oxygen induced retinopathy (OIR) group (n=136). Among room-air group, there were 18 A2A knock-out (KO) mice (KO subgroup) and 24 C57BL/6 mice as wide type (wide type subgroup). OIR group were divided into OIR control subgroup (n=48), A2A-OIR subgroup (n=24) and Caffeine-OIR subgroup (n=64). The retinal neovascularization of OIR group was induced by oxygen. The pathological neovascularization was determined by retinal sections. Fluorescent quantitative polymerase chain reaction (PCR) was used to measure the mRNA expression of A2A and vascular endothelial growth factor (VEGF). 0.1, 0.3, 1.0 g/L Caffeine was dissolve in drinking water of lactating females in Caffeine-OIR subgroup, non-perfusion areas of retina in mice at the age of 0 - 17, 0 - 7, 7- 17, 7-12, and 12- 17 days were analyzed in different dosage and when the dosage as 1.0 g/L. Results Compared with OIR control subgroup, the retinal non-perfusion areas and the numbers of endothelial cell nuclei breaking through the internal limiting membrane in A2A- OIR subgroup were reduced significantly (t=7.694, 7.747;P＜0.001). Compared with wide type subgroup, the level of A2A and VEGF mRNA in OIR control subgroup increased significantly (t=4.036, 2.230;P＜0.05). Compared with OIR control subgroup, the level of VEGF mRNA in A2A- OIR subgroup decreased significantly (t=3.122,P＜0.01). Compared with OIR control subgroup, the retinal non-perfusion areas in mice at the dosage of 0.1 and 1.0 g/L (t=2.397, 4.533) and at the age of 0 -17, 0 -7 days when the dosage as 1.0 g/L (t=4.070, 2.399) were reduced significantly (P＜0.05). Conclusions The expression of adenosine A2A receptor increases in oxygen-induced retinal pathological neovascularization. Adenosine A2A receptor may regulate the expression of VEGF. A2A receptor inactivation can inhibit oxygen-induced retinal pathological neovascularization.
Objective To investigate the effect of peptidoglycan (PGN) on the secretion of pro-inflammatory cytokines by dendritic cells (DCs) and the regulation of T helper 17 (Th17) responses in experimental autoimmune uveitis. Methods Bone marrow cells from naive mice were cultured with granulocyte macrophage-colony-stimulating factor and interleukin (IL)-4 to induce DCs. DCs cultured for six days were randomly divided into two groups: PGNtreated group and control group. The DCs in PGNtreated group were stimulated with PGN and the same volume of phosphate buffered saline was added to the DCs as control group. The relative mRNA expression levels of IL-23, tumor necrotic factor alpha; (TNF-alpha;), IL-6,IL-1beta;were measured by real-time reverse transcriptase polymerase chain reaction (RT-PCR). Peptide fragment of interphotoreceptor retinoidbinding protein (IRBP1-20)specific T cells, which were isolated from the spleen and draining lymph nodes of C57BL/6 mice immunized with IRBP1-20 peptide fragments 13 days earlier, were co-cultured with PGN-treated or untreated DCs, respectively. Total RNA from T cells cocultured for two days were isolated and the relative expression of retinoic acid receptor-related orphan receptor gamma;t (ROR-gamma;t), IL-17, T-box expression in T cells (T-bet), interferon gamma; (IFN-gamma;) mRNA were detected by realtime RT-PCR. On the second, the fifth and the seventh day, the cocultured T cells were analyzed by flow cytometry to detect the percentages of IFN-gamma;, IL-17 positive cells. Results The real-time RT-PCR results revealed that the level of IL-23, IL-1beta;, IL-6, TNF-alpha; mRNA from PGNstimulated DCs were significantly increased compared to the control group (t=-14.363, -5.627, -3.85, -28.151; P＜0.05). The level of RORgamma;t, IL-17 mRNA from the T cells cocultured with PGN-stimulated DCs were greatly increased compared with the control group (t=-5.601, -19.76；P＜0.05). However, the level of T-bet, IFN-gamma; mRNA from the T cells cocultured with PGNstimulated DCs were significantly decreased compared with the control group (t=4.717, 11.207; P＜0.05). Data of flow cytometry showed that at two days, five days, seven days after cocultured with PGN-treated DCs, the percentages of IL-17 positive T cells were increased compared to the control group (t=-2.944, -3.03, -4.81; P＜0.05), and the percentages of IFN-gamma; positive T cells had no remarkable change (t=-1.25, -0.18, -2.16; P＞0.05). Conclusion PGN can promote the secretion of Th17-related cytokines by DCs, which favors proliferation and differentiation of Th17 in experimental autoimmune uveitis.
Objective To observe the effect of high glucose on the expression of activating transcription factor 4 (ATF4) in cultured retinal Muuml;ller glia cells. Methods The retinal tissue of Sprague-Dawley (SD) rats was collected, and Muuml;ller cells were isolated and cultured. The glial fibrillary acidic protein (GFAP) and glutamine synthetase (GS) of Muuml;ller cells were identified by streptavidin-biotin-peroxidase complex. Cultured rat Muuml;ller cells were divided into control group (5.5 mmol/L glucose), group A (20 mmol/L glucose), group B (30 mmol/L glucose) and group C (40 mmol/L glucose). ATF4 protein expressions in Muuml;ller cells of four groups were measured by Western blot four days after cultured. Results GFAP and GS expressed in more than 95% of Muuml;ller cells. Over 95% of Muuml;ller cells of group A, B and C were positive for GFAP and GS. Western blots indicated that ATF4 protein in group A, B and C increased obviously compared with the control group (q=0.293, 0.754,0.484;P＜0.05). Conclusion High glucose can increase the expression of ATF4 protein and cause endoplasmic reticulum stress in retinal Muuml;ller glia cells in vitro.
Objective To observe the proportion changes of CD4+CD25+FOXP3+ T cells in peripheral blood of patients with VogtKoyanagiHarada disease (VKH) before and after one month of treatment. Methods he peripheral blood samples from 15 patients with VKH disease before and after one month of treatment by glucocorticoid, and from 15 healthy volunteers were collected，and lymphocytes were separated from them. CD4+CD25+ regulatory T cells were labeled by antibodies of cell surface marker CD4、CD25 and transcription factor FOXP3. The proportion of CD4+CD25+FOXP3+ T cells were detected by flow cytometry. Results Before the treatment, the percentage of CD4+CD25+FOXP3+ T cells in periphery blood was（0.30plusmn;0.19）% of CD4+ cell in VKH patients, and（1.41plusmn;0.52）% in control group, the difference was statistically significant（t=7.665,Plt;0.01）; after one month of treatment, the VKH patients group was（1.28plusmn;0.54）% which close to the control group. However there were two patients whose CD4+CD25+ T cells increased extraordinarily after one month of treatment. Conclusions The proportion of CD4+CD25+ FOCP3+ T cells in periphery blood in VKH patients were lower than control group obviously before treatment, but were close to control group after treatment. Those results indicated that VKH diseases may be associated with the decreased proportion of CD4+CD25+ regulatory T cells.
Objective To observe the characteristics of morphosis parameter of the optic disc of physiologic large cup. Methods 100 eyes with physiologic large cup and 74 eyes with normal cup were examined by Heidelberg Retina Tomograph (HRT ). The differences of morphosis parameters between two groups were analyzed comparatively on disc area (DA), cup volume (CV), cup/disc area ratio (C/DR), rim area (RA), cup volume (CV), rim volume (RV), mean cup depth (MeCD), maximum cup depth (MxCD), cup shape measure (CSM), height variation contour (HVC), mean retinal nerve fiber layer thickness (mRNFLt), and retinal nerve fiber layer cross-section area (RNFLcsa). The characteristics of the inferior, superior, nasal, and temporal quadrants of the physiologic large cups were analyzed. Results DA, CA, C/DR, CV, MeCD, CSM （P=0.00, respectively）and MxCD （P=0.04）were significantly larger in eyes with physiologic large cup than in eyes with normal cup. RA, RV, HVC, mRNFLt, RNFLcsa （P=0.00, respectively) were significantly smaller in eyes with physiologic large cup than in eyes with normal cup. The temporal quadrant of RV of the physiologic large cup is the narrowest. RNFLcsa decreased as the fol lowing order: superior, inferior, nasal, and temporal（P＜0.05）. Conclusions Mo rphosis parameter of the optic discs of physiologic large cup has its own repres entation on characteristics. Compared to normal cups, physiologic large cups had larger discs but smaller mRNFLt. The nasal quadrant of DA was larger than the i nferior quadrant. （Chin J Ocul Fundus Dis，2008，24：213-216）
Objective To observe the morphological changes of dendrite and soma in retinal ganglion cells (RGCs) which subsisted in early diabetic rats. Methods The RGCs of 3-months-course diabetic rats and coeval normal rats were marked by gene gun techniques. To collect RGCs photographs by Leica microscope with Z axis and CCD camera；to observe the changes of diameter, variance of structural features in dendritic field and somata after classification which according to the size and morphology. Thy-1 antibody marks on the retinal RGCs, taking a photograph under fluorescent microscope, counting the changes of retinal RGCs density in early diabetic rat. Results In three-month diabetic rats,the density of retinal RGCs was decreased obviously. Morphological changes of RGCs in the dendritic fields were observed with gene gun technique. There was no severe variation in all kinds of the bole of cell dendrite, in which some only showed crispation partially and sparseness also twisting in the dendritic ramus. The mean diameter of dendritic field and soma in class A of diabetic rats was (401plusmn;86) mu;m, the mean diameter of dendritic field in control group was (315plusmn;72) mu;m,compared with each other, there is statistically significant differences (t=21.249，Plt;0.001); the mean diameter of soma in class A of diabetic rats was (24plusmn;6) mu;m, the mean diameter of soma in control group was (22plusmn;5) mu;m, compared with each other, there is no statistically significant differences (t=0.927，Pgt;0.05); the mean diameter of dendritic field and soma in class B of diabetic rats were (170plusmn;36)、(14plusmn;2) mu;m respectively, in control group were (165plusmn;36)、(16plusmn;2) mu;m, the mean diameter of dendritic field and soma in class C of diabetic group were(265plusmn;78)、(17plusmn;5) mu;m respectively, in control group were (251plusmn;57)、(17plusmn;4) mu;m , compared with each other, there are on statistically significant differences(t=1.357，0.798，0.835，1.104，Pgt;0.05). Conclusions In short-term diabetes, the survived RGCs show good plasticity in adult diabetic rats, especially in class A. The changes of dendrites were more sensitive than the soma, which could be the leading index of the morphologic changes of RGCs in the early stage. The good plasticity showed by the RGCs and the time window from changing in dendrite to cell death provide us many evidences not only for the research but also for the nerve protection in clinic. (Chin J Ocul Fundus Dis,2008,24:249-254)
The pathogenesis of diabetic retinopathy (DR) is more complex. For the upstream of traditional pathogenesis, to looking for unifying mechanism theory which proposed in foundation of common promoters and the latest view of DR may be the result of chronic inflammation. Both of them provide the basic and clinical theraby of DR with new direction. Therefore, there are many related issues still needs to intensive study. （Chin J Ocul Fundus Dis，2008，24：237-239）
Objective To observe the influence of the expression of CD18 on the neutrophile and the leukocyte adhesion to retinal vascular endothelium by hypoxia-inducible factor-1 alpha (HIF-1alpha;) in early diabetic retinopathy rats. Methods Male Sprague-Dawley rats received intraperitoneal injection of streptozotocin to induce diabetes model. 18 diabetic rats were divided into 3 groups randomly after 2 months of diabetes induction, including diabetic group (group B), HIF-1alpha; anti-sense oligonucleotides (ASODN) injection group (group C) and HIF-1alpha; sense oligonucleotides (SODN) injection group (group D), the age and weigh matched health rats were chosen as control group (group A), with 6 rats in each group. Then group A and B rats received 5% glucose solution caudalis veins injection, group C and group D rats received HIF-1alpha; ASODN and HIF-1alpha; SODN caudalis veins injection, respectively(025 mg/kg).The level of CD18 on the neutrophil isolated from the peripheral blood was measured by flow cytometry. Retinal leukostasis was quantified with acridine orange leukocyte fluorography. Results The percentage of CD18 positive neutrophil cell was（44.93plusmn;3.60）% in group B，（18.66plusmn;1.52）% in group A，（31.66plusmn;4.72）% in group C，（51.00plusmn;5.66）% in group D. Compared with each other groups，the differences are statistically significant （F=42.46, Plt;0.001）. The number of positive staining cells of retinal leukocyte was （46.16plusmn;10.68）in group A，（133.83plusmn;20.43）in group B,（99.83plusmn;9.28）in group C，（121.33plusmn;10.23） in group C. Compared group B with group C，the number of positive staining cells raised about 2.89 times；compared group B with group C and D，the differences are statistically significant （P=0.12，95% confidence interval -3.69~28.69）. Conclusions In vivo, HIF-1alpha; can decreased the expression of CD18 on neutrophils from diabetic ratsprime; peripheral blood and the collection of retinal leukostasis in the diabetic animals. HIF-1alpha; may serve as a therapeutic target for the treatment and/or prevention of early diabetic retinopathy. （Chin J Ocul Fundus Dis，2008，24：268-271）