ObjectiveTo observe the effect of epinephrine in intraocular irrigation solution on retinal vascular caliber and macular thickness. MethodsA prospective control study. 32 eyes of 32 patients with macular hole who underwent vitrectomy were enrolled in this study. The patients including 14 males (14 eyes) and 27 females (18 eyes), with the average age of (64.0±4.5)years. Uncorrected visual acuity, corrected visual acuity, slit lamp biomicroscopy, indirect ophthalmoscopy, fundus color photography and optical coherence tomography were performed in all patients. Retinal vascular caliber located in 0.5-1.0 disc diameter from optic disk was measured from digital fundus photographs and summarized as central retinal artery (CRAE) and vein (CRVE) equivalents in all eyes at baseline and at the 1 month, 3 months follow-up visit. The macular thickness is the distance from retinal interface of inner plexiform layer to retinal pigment epithelium layer. The macula was divided into inner ring ( < 3 mm) and outer ring (3-6 mm) according to the distance from the fovea. The patients were divided into experiment group (include epinephrine in intraocular irrigation solution, 1:1000) and control group (without epinephrine in intraocular irrigation solution), 16 eyes in each group. The difference of CRAE and CRVE between two groups was not significant (P > 0.05). The difference of macular thickness between inner ring and outer ring was not significant (P > 0.05). The average follow-up was 3.5 months. CRAE, CRVE and macular thickness in inner ring and outer ring before and 1 month, 3 months after surgery were comparatively analyzed. ResultsThe differences of CRAE and CRVE before and 1, 3 months after surgery both in experiment group (tCRAE=0.322, 0.148; tCRVE=0.317, 0.005) and control group (tCRAE=0.226, 0.137; tCRVE=0.284, 0.151) were not significant (P > 0.05). The differences of CRAE (t=0.624, 0.424) and CRVE (t=0.015, 0.041) between experiment group and control group also were not significant (P > 0.05). The differences of macular thickness in inner ring and outer ring before and 1, 3 months after surgery both in experiment group (tinner=0.322, 0.148;touter=0.317, 0.005) and control group (tinner=0.226, 0.137;touter=0.284, 0.151) were not significant (P > 0.05). The differences of macular thickness in inner ring (t=1.568, 0.373) and outer ring (t=-1.697, 0.536) between experiment group and control group also were not significant (P > 0.05). ConclusionEpinephrine (1:1000) in intraocular irrigation solution has no effect on retinal vascular caliber and macular thickness in patients with macular hole.
ObjectiveTo evaluate the protective effect of estrogen on survival of retinal ganglion cells (RGCs) after transient retinal ischemia-reperfusion (RIR) in rats.MethodsRIR was induced in 60 ovariectomized adult rats (OVX) by increasing intraocular pressure via an intracameral catheter. All of the rats were divided into two groups randomly: in experimental group, the rats underwent a subcutaneous injection with 17β-estrodiol（100 μg/kg） 2 hours before retinal ischemia; and in the control group, saline water was injected correspondingly. The number of RGCs and the thickness of the inner retinal layers were mesured by HE staining method before and 12, 24, 48, and 72 hours after reperfusion. TdT-mediated biotin-dUTP nick end labelling (TUNEL) staining technique was used to examine the apoptosis of RGCs.ResultsTwenty-four and 48 hours after reperfusion, the number of apoptotic cells in experimental group was obvious lower than that in the control group(Plt;0.05), and the number of RGCs in experimental group was higher than that in the control group(Plt;0.05).ConclusionEstrogen can protect retinal neurons from transient RIR in ovariectomized rats.(Chin J Ocul Fundus Dis, 2005,21:177-179)
ObjectiveTo observe the stoichiometry of vascular endothelial growth factor receptor 2 (VEGFR2) on the retinal vascular endothelial cell membrane by single-molecule fluorescence imaging.MethodsRhesus monkey retinal vascular endothelial cells (RF/6A) were divided into blank control group (normal culture) and plasmid transfection group [transfected with VEGFR2-green fluorescent protein (GFP) recombinant plasmid]. The expression of GFP in the plasmid transfected group was observed by confocal microscope, and the expression of VEGFR2 in the cells was detected by real-time fluorescent quantitative polymerase chain reaction (qPCR) and Western blot. The fluorescence intensity distribution and bleaching steps of single VEGFR2-GFP molecule on the cell membrane were recorded by single-molecule imaging. The distribution of fluorescence intensity and the number of fluorescence bleaching steps of GFP were recorded.ResultsGFP green fluorescence was observed in the transfected cells 12 hours after transfection. qPCR results showed that the expression of VEGFR2 and GFP mRNA in the plasmid transfected group was significantly higher than that in the blank control group (t=11.240, 12.330; P＜0.001, 0.001). Western blot results showed that the expression of VEGFR2 protein in the plasmid transfected group was significantly higher than that in the blank control group (t=8.346, P＜0.01). The results of single-molecule imaging showed that the fluorescence intensity distribution of VEGFR2-GFP on the surface of RF/6A cell membrane without ligand stimulation was bimodal, in which monomer and dimer were 86.0% and 14.0% respectively. By counting the steps of GFP fluorescence bleaching, the proportions of receptor monomer, dimer, trimer, and tetramer were 81.4%, 12.9%, 5.5%, and 0.3% respectively.ConclusionIn the absence of ligands, VEGFR2 coexists in the form of monomers and dimers on the surface of RF/6A cell membrane, and monomers are dominant.
OBJECTIVE :To investigate ocular vessel flow velocity in normal eyes by color Doppler imaging(CDI). METHODS: Ninety people (180 normal eyes)had flow velocity measured by CDI in three vessels,ophthalmic artery (OA),central retinal artery(CRA) and posterior ciliary artery (PCA),and the relations between velocity and affecting factors were analysed. RESULT: The diastolic and systolic velocity of OA,CRA and PCA were (31.7plusmn;10.9)cm/s, (7.2plusmn;2.6)cm/,s, (10.2plusmn;3.4)cm/s, (2.8plusmn;1.2)cm/s and (11.3plusmn;3.6)cm/s, (3.2plusmn;1.4)cm/s respectively. The velocity of OA had possitive correl-ativity with RBC,HCT and negative correlativity with age,while it revealed no relationship with sex, laterality of right or left eye,and normal IOP. CDI of ocular vessels in normal eyes is a triangle with three peaks and double sunkens in its frenquency spectum, revealed highly resistant,and both the pulsatility and resistive indexes were relatively high. The width of each frequency band basically was the same,and there was no obvious frequency window. CONCLUSION: The ocular application of CDI might lay the foundation of a comparatively comprehensive knowledge of the ocular hemodynamics. （Chin J Ocul Fundus Dis,1997,13: 99-101）
The retinal vessel changes are the primary and major features of retinal vascular diseases. The retinal vessel is part of systemic vessels with its own characteristics to sustain normal retinal function. These basic characteristics are important to the correct understanding and proper treatment of retinal vascular diseases. Always keep in mind that the retinal vessels is one part of the systemic vascular system, thus retinal vascular diseases may have systemic etiology, and systemic drug administration may have a profound effects to the whole body. However retinal vascular system also has its own structural and functional characteristics, thus retinal vascular diseases are also different from the systemic diseases. Finally the main function of retinal vascular network is to maintain the neuro-retinal function, thus we should balance the vision protection and treatments against abnormal retinal blood vessels. Over-treatments may damage the retinal vision.
ObjectiveTo measure and analyze the oxygen saturation and retinal blood vessel diameter in the eyes of patients with convalescence Vogt-Koyanagi-Harada (VKH) syndrome. MethodsIn this cross-sectional study, 28 eyes of 14 patients with convalescence VKH syndrome (VKH group) and 20 eyes of 10 healthy subjects (control group) were enrolled. The oxygen saturation and retinal blood vessel diameter were detected by spectrophotometric oximetry unit. Retinal images were collected using filters with wavelengths of 570 nm and 600 nm in the darkroom by the same technologist and then the fused image was obtained. The oxygen saturation of retinal vessels was marked in different colors. The measurement was repeated 2-3 times for each patient, then take an average. A top-quality image in each eye was selected to detect the oxygen saturation and diameter of retinal vessel which located in 1.5-3.0 disc diameter from the optic disc. Image analysis and data acquisition were completed by another technologist. ResultsRetinal venous oxygen saturation was (54.34±8.05)% in the VKH group and (60.07±7.91)% in the control group. The former was lower than the latter, the difference was significant (t=2.443, P=0.017). The mean diameter of retinal arteries was (102.8±18.1) μm in the VKH group and (112.9±19.8) μm in the control group. The former was smaller than the latter, the difference was significant (t=2.406, P=0.018). There was no significant difference of the mean diameter of retinal veins, oxygen saturation of retinal arteries and the arterial-venous difference between two groups (t=-0.330, 0.804, -0.631; P=0.743, 0.403, 0.536). ConclusionsRetinal venous oxygen saturation and the mean diameter of retinal arteries are significantly decreased in patients with VKH syndrome. There is no significant difference of diameter of retinal veins, oxygen saturation of retinal arteries and the arterial-venous difference between VKH syndrome patients and healthy subjects.
The activities and distributions of succinate dehydrogenase(SDH),malic dehydrogenase(MDH),lactic dehydrogenase(LDH),acid phosphatase(ACP) and alkaline phosphatase(AKP) in retinal vessels were studied and observed with enzymatic histochemical techniques. The retinal vessels showed a b LDH activity, moderate SDH and MDH activity. The dehydrogenase activity described above was evenly and equally distributed in the microvasculature between arterioles and venules, and was the best in arteries. AKP showed predominant activity in the endothelial cells of capillaries and arterioles which were stained bly. No activity for ACP observed in the retinal vessels. The observations above indicate that the retinal vessels are metabolically active and have a great capacity for glycolysis. （Chin J Ocul Fundus Dis,1992,8:6-9）
0bjective To explore the effect of endothelin(ET)、nitrioxide (NO) in plasma on retinopathv in the pregnancy-induced hypertension(PIH)． Methods The 1evel of ET and NO in plasma of 75 cases of in-patient women with PIH and 20 cases of women with the full terms and normal pregnancy before and after delivery was determined by radioimmunoassay．The retinopathy of the patients with FIH before and after delivery was detected by appointed doctor．The levels of ET and N0 in both groups were compared and the relationship between ET and N0 in plasma and the retinopathy before and after the delivery was detected.Results The levels of ET[(145．oo±54.41)ng／L] in serious PIH patients were much higher than that in the control[(81．50±43．80)ng／L]，the minor[(85.30±33.33)ng／L]and middling PIH group[(90．20±39．25)ng／L]．The levels of ET in plasma before and after pregnancy were not changed in PIH patients [(118．70±33．44)ng／L]，but were higher than that in the control group. The levels of plasma NO in serious[(87．56±35．58)ng／L]and middling[(78.11±28.96)ng／L] PIH group were both higher than that in the control group[(46．70±32.64)ng/L]，and the levels in minor(52．56±28．35)ng／L]and middling PIH group were lower than that in the serious PIH group.The level of N0 in plasma of PIH patients after the delivery was much lower than that before the delivery，while higher than that in the control．The positive correlation between levels of ET and NO and retinopathy was found in PIH patients.Conclusions The 1evels of plasma ET and N0 in PIH patients are related to the extent of the disease，and the level of ET in plasma is highly related to the retinopathy in PIH patients, ET and NO might be played an important role in pathogenesis of retinopathy and ET might be a good index in reflecting the rank of retinopathy in PIH.(Chin J Ocul Fundus Dis，2004，20：12-15)
Objective To investigate the effect of hypoxia on the exp ression and function of integrin receptor αvβ3 of bovine retinal vascular endotheliocytes. Methods Bovine retinal vascular endotheliocy tes in the culture dishes coated by vitronectin was put into the normal and hypoxemic condition, respectively. Enzyme linked immunosorbent assay and cell adhesion analysis were used to detect the expression and function of integrin receptor αvβ3 in bovine retinal vascular endotheliocytes, respectively. Results Under the condition of hypoxia, the expression of αvβ3 increased gradually, and reached the peak at the 48th hour. The expression of αvβ3 at the 60th and 72nd hour in hypoxia group was higher than that in the normal group. Bovine retinal vascular endotheliocytes absorbed more Vn of extra-cellular matrixes (ECM) after cultured under hypoxemic condition for 24 hours.Conclusion Hypoxia may up-regulate the expression of αvβ3, which promote the adsorbability of endotheliocytes.(Chin J Ocul Fundus Dis，2004，20：360-363)
ObjectiveTo observe the MiSeq sequencing analysis results of fulvic acid (FA) intervention in hypoxia-induced human retinal microvascular endothelial cell (hRMEC) gene expression profile.MethodshRMEC were cultured in vitro and divided into the hypoxia group (hypoxia treatment) and the FA intervention group (FA intervention after hypoxia). The MTT colorimetric method was used to detect the influence of different concentrations and different modes of FA on hRMEC activity. The optimal concentration of FA was chosen. RT-PCR was used to investigated the effect of FA on hypoxia-induced intercellular adhesion molecule-1 (ICAM-1), IL-1β, IL-4, IL-6, IL-6, IL-8, IL-10, MMP-2, TNF-α, TNF-β, other inflammatory factors in hRMEC, and inflammation-related factors mRNA expression. Cells in the hypoxia group and FA intervention group in the logarithmic growth phase were collected. MiSeq sequencing technology was applyed to complete the whole transcriptome sequencing of the two groups of cells, biological data were obtained, and the differentially expressed miRNA were analyzed on this basis. Gene annotation (GO) functionally significant enrichment analysis and Kyoto Encyclopedia of Genes and Genome (KEGG) pathway significant enrichment analysis were used to analyze the functions and signal pathways of differential miRNAs. The expression of inflammatory factors and inflammation-related factors were compared between groups. The expression level of the corresponding miRNA in the cell was regulated by miRNA mimic, and its effect on cell function was observed, so as to judge the effect of the miRNA.ResultsDifferent concentrations and different modes of action of FA had no effect on the cell viability of hRMEC. The mRNA expression of ICAM-1, IL-1β, IL-6 and TNF-β in the hypoxia group hRMEC were significantly up-regulated compared with the normal group, and the difference was statistically significant (t=3.426, 6.011, 5.282, 6.500; P=0.027, 0.004, 0.006, 0.003); the mRNA expression of ICAM-1, IL-6, TNF-α and TNF-β in the FA intervention group hRMEC was significantly lower than that of the hypoxia group, and the difference was statistically significant (t=9.961, 3.676, 3.613, 3.387; P=0.001, 0.021, 0.023, 0.028). There were 14 differentially expressed miRNAs between the hypoxia group and the FA intervention group, of which 9 were up-regulated genes and 5 were down-regulated genes. The predicted target genes of 4 differential miRNAs (hsa-miR-1285-3p, hsa-miR-30d-3p, hsa-miR-3170, hsa-miR-7976) were all ICAM-1. The results of significant enrichment analysis of GO function showed that the functions of differential genes were mainly enriched in the process of cell development, cell differentiation and single organism development. Significant enrichment analysis of the KEGG pathway showed that the differential miRNA expression was highly enriched in the proteoglycan pathway and the cytokine-cytokine receptor interaction pathway in cancer, and the arachidonic acid metabolism pathway and the amphetamine pathway were the more obvious differential expressions.ConclusionFA may affect the expression level of downstream ICAM-1 mRNA by regulating the expression of multiple miRNAs, thereby affecting the inflammatory state of cells after hypoxia-stimulated hRMEC.