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 changes of ocular hemodynamics in patients with retinal vein occlussion(RVO)． Methods The hemodynamic parameters(PSV,EDV,PI,Vmax)of central retinal artery(CRA)and central retinal vein(CRV)were measured in the involved eyes(n=48) with RVO and the contralateral clinically healthy eyes(n=39) and in the control eyes(n=40) by color Doppler imaging (CDI)(ATLHDI3000). Results Peak systolic velocity (PSV) and end diastolic velocity (EDV) were significantly lower in the CRA of involved eyes and clinically healthy eyes of patients with RVO compared with control eyes,and pulsatility index(PI)was significantly higher in the CRA of involved eyes of patients with RVO compared with control eyes．PSV were significantly lower in the CRA of involved eyes of patients with RVO compared with their clinically healthy eyes．Pulsatility index(PI)was significantly higher in the CRA of involved eyes of patients with RVO compared with their clinically healthy eyes．Maximun vein velocity (Vmax) was significantly lower in the CRV of involved eyes and clinically healthy eyes of patients with RVO compared with control eyes． Conclusion The changes of hemodynamics in CRA,CRV of involved eyes of patients with RVO may invade their clinically healthy eyes．CDI may be helpful to early diagnosis for RVO． (Chin J Ocul Fundus Dis,1998,14:111-113)
Objective It has been shown that pigment epitheliumderived factor (PEDF) is an effective anti-apoptosis agent on several kinds of cells of the central nervous system.This study aimed to evaluate the effect of PEDF on pressure induced retinal ischemia in a rat model. Methods Retinal ischemia was induced by increasing the intraocular pressure to 110 mm Hg for 45 minutes via an intracameral catheter.Ten microlit ers (0.1 mu;g/mu;l) PEDF was injected into the vitreous of 4 eyes of each group im mediately after reperfusion and 4 additional eyes received only normal saline as vehicle controls.The animals were euthanized at 2 or 7 days after reperfusion.T he effect of PEDF on retinal degeneration was assessed by measuring the thicknes s of the inner retinal layers (MTIRL) and counting the retinal ganglion cells (R GC) on plastic embedded retinal sections. Results The MTIRL and the RGC counting in eyes treated with intravitreal PEDF were significantly higher than those in vehicle controls (118.1plusmn;5.0) mu;m vs(94.9plusmn;3.0) mu;m (Plt;0.05);(6.0plusmn;1.0) cells/100 mu;m vs (4.5 plusmn;0.5) cells/100 mu;m (Plt;0.05) 7 days after reperfusion,respectively． Conclusion Intravitreal administration of PEDF can ameliorate an ischemiareperfusion retinal injury and may be useful to prevent neuronal degeneration in the inner retina. (Chin J Ocul Fundus Dis, 2001,17:138-140)
Objective To investigate the spatial and temporal regulation effect of VEGF on human fetal retinal vascularization and angiogenesis. Methods The posterior segmental retinas from 54 human fetuses of the 9th week to the 40th week were studied by immunohistodhemistry standing for the expressions of VEGF and PCNA. Results 1. The distribution of VEGF espression was spiking and the peaks were during the 9th-13th and around the 26th week. 2. PCNA immunoreactivity was localized in spindle cells and vascular endothelial cells. The expression level was fluctuated during the developmental process. The peaks were during the 9th-13th and around the 21st week. In these periods, the spindle cells kept proliferating and differentiating, and remodelled subsequently to form the inner side retinal vessels. From the 26th or 34th week, the PCNA immununoreactivity is fully expressed in the vascular endothelial cells of the inner and outer margin of inner nuclear layer(INL) and kept to full terms. 3. Significant positive correlation were shown between the content of VEGF in the retina and that of PCNA in spindle cells and vascular endothelial cells(r=0.736,p＜0.01). Conclusion VEGF was positively involved in modulating human fetal retinal vascularization and angiogenesis. (Chin J Ocul Fundus Dis,1999,15:12-15)
ObjectiveTo explore the effects of transthyretin (TTR) on biological behavior of retinal microvascular epithelial cell (RMVEC). MethodsRMVEC was cultured in medium with 0 μmol/L and 4 μmol/L TTR. The proliferation, migration and healing abilities (0, 24, 48 hours) of RMVEC with different concentrations of TTR were measured by methyl thiazol tetrazolium (MTT) assay, transwell assay and scarification test. ResultsMTT assay shows that RMVEC with the concentrations of 4 μmol/L TTR [absorbance (A) value=0.17±0.02] glows faster than with the concentrations of 0 μmol/L TTR (A value=0.40±0.03), the difference was statistically significant (t=15.47, P=0.000 1). The transwell assay shows RMVEC with the concentration of 4 μmol/L TTR [(140±7) cells] migrants faster than RMVEC with the concentration of 0 μmol/L TTR [(227±14) cells], the difference was statistically significant (t=5.44, P=0.000 6). The scarification test shows that the RMVEC with the concentration of 4 μmol/L TTR [(134.4±45.4) μm] heals faster than the RMVEC with the concentration of 0 μmol/L TTR [(330.0±23.1) μm], the difference was statistically significant (t=8.25, P<0.01). The cells in 48 hours and 4 μmol/L group were healed completely, but not healed in 0 μmol/L group. ConclusionTTR can promote the proliferation, migration and healing abilities of RMVEC.
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)
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.
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.
Objective To observe the expression of p53, bcl-2 genes, vascular endothelial cell growth factor（VEGF）, basic fibroblast growth factor（bFGF）, insulin-like growth factor-I （IGF-I）, and the receptors of these factors of retinal vascular endothelial cells (VECs) of 1- to 20-week diabetic rats, and the relationship between the expressions and cell cycle arrest.Methods Retinal sections of diabetic rats induced by alloxan were immunohistochemically stained and observed by light microscopy (LM) and electron microscopy (EM). Dot blotting and Western blotting were used to determine the expression of mRNA, proteins of p53 and bcl-2. Results Under LM, immunohistochemical positive expression of p53 and bcl-2 were found on the vessels of ganglion cell layer and inner nuclear layer of retinae of 8- to 20-week diabetic rats; under EM, these substances were observed depositing in VECs. The retinal VECs also expressed VEGF, bFGF, IGF-I and their receptors. There was no positive expression of other cell types in these retinae, all cell types of retinae in control group, or all cells of retinae of diabetic rats with the course of disease of 1 to 6 weeks. The result of dot blotting revealed that retinal tissue of 20-week diabetic rat expressed p53 and bcl-2 mRNA, and the result of Western blotting revealed that they also expressed p53 and bcl-2 proteins. But retinal tissues of control group did not. Positive expression of bax was not found in the retinae in control group or 1- to 20-week diabetic rats. Conclusion p53, bcl-2 may introduce cell cycle arrest of VECs of retinae in 8- to 20-week diabetic rats. High glucose might stimulate the expression of VEGF, bFGF, IGF-I and their receptors, and the growth factors may keep VECs surviving by self-secretion. (Chin J Ocul Fundus Dis，2003，19：29-33)