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)
Purpose To clarify the relationship between diabetic retinopathy (DR) and maculopathy (DM) and explore the clinical implication of independent graduation of DM. Methods Fundus fluorescein angiography and routine ophthalmological examination were performed on 582 cases of diabetes.Their ocular fundi and macular impairments were graded. Results In general,the severity of diabetic macular impairment was accompanied by retinal involvement,but discrepancy existed between DM and DR.Degree I DM occurred in 5.4% (16/294) among cases without DR,in stage IV DR,degree Ⅲ DM accounted for the most part ,54.5% (116/213).There were still 5.1% (2/39) cases without DM in stage Ⅴ DR. Conclusion The degree of the macular lesions in DM is often not in parallel with the gradation of general affections in retinal tissue other than in macular region in DR,therefore,independentg radation of diabetic maculopathy has its clinical significance for choosing the optimal period of treating maculopathy and preserving the macular function. (Chin J Ocul Fundus Dis,2000,16:153-154)
Objective The bone marrow mesenchymal stem cells (BMSCs) have the capacity to differentiate into insul in-producing cells (IPCs) in vitro. However, low differentiation efficiency and poor maturity are the main obstacles. To investigate the feasibil ity of BMSCs differentiation into IPCs in diabetic pancreatic microenvironment of pigs. Methods BMSCs were isolated and purified from the bone marrow of a 4-week-old male pig. Fifteen female pigs (aged 8 to 10 weeks, weighing 8 to 10 kg) were randomly divided into 3 groups: normal control group (group A, n=5), diabetic control group (group B, n=5), and BMSCs transplanted group (group C, n=5). The pigs of groups B and C were treated by auris vein injections of styeptozocin and alloxan for 3 days to induce diabetes mell itus (DM) model, whose blood glucose level 2 days all greater than 17 mmol/L was successful DM model. A total of 1.1 mL of the 3rd passage BMSCs labeled with enhanced green fluorescent protein (EGFP), with cell density of 5 × 107/ mL, were injected into subcapsular pancreas of group C at multi ple points, normal saline at the same dosage into those of groups A and B. After 30 days of monitoring blood glucose, the histological analysis of islet number and size were done; the immunofluorescence staining was used to detect the protein expression of insul in in the new-formed islets. The EGFP+ cells were collected from the sections using laser-capture microdissection; RT-PCR was used to detect insulin mRNA and pancreatic and duodenal homeobox factor 1 (PDX1) mRNA expressions from EGFP+ cells, and the insul in and sexdetermining region of the Y chromosome (SRY) genes were detected by fluorescence in situ hybridization (FISH). Results The blood glucose level decreased significantly in group C when compared with that in group B from 18 days and gradually decreased with time (P lt; 0.05). The histological observation showed that the number of islets was increased significantly in group C when compared with that in group B (10.9 ± 2.2 vs. 4.6 ± 1.4, P lt; 0.05), and there was no significant difference when compared with that in group A (10.9 ± 2.2 vs.12.6 ± 2.6, P gt; 0.05). The size of new-formed islets in group C was significantly smaller than that in group A [(47.2 ± 19.6) μm vs. (119.6 ± 27.7) μm, P lt; 0.05]. The immunofluorescence staining showed that new-formed islets of group C expressed insulin protein. RT-PCR showed that the microdissected EGFP+ cells of group C expressed insulin mRNA and PDX-1 mRNA. FISH showed that the new-formed islet cells of group C contained SRY gene in Y chromosome and insulin double positive cells. Conclusion BMSCs can differentiate into IPCs in diabetic pancreatic microenvironment of pigs.
ObjectiveTo investigate relationship between ultrastructural changes and expression of basic fibroblast growth factor of diabetic retinopathy in rats.MethodsDiabetes was induced in rats with a single injection of streptozotocin (STZ) and divided into normal control group and 1- , 3- and 5- month diabetes group. The paraffin slide was observed by in-situ hybridization and immunohistochemistry, and retinal ultrastructure was examined by transmission electron microscopy.ResultsNo change of retinal ultrastructure was found in the control group. Different degrees of ultrastructure lesion were found in 1-month diabetic rats with fragmental increase of thickness of basement membrane, swelling of endothelial cells and obvions fingerlike processes in the capillary cavity, disconcentration of heterochromatin both in endothelium and pericyte, and swelling and degeneration of mitochondrion. The edema of endothelial cells of 3-month diabetic rats was more serious than that of 1month ones, and the capillary cavity was nearly occluded. In 5-month diabetic rats, the basement membrane was unevenly thickened, or obviously split. The positive rate of in-situ hybridization in 3-month diabetic rats was 77.8% while the positive rate of immunohistochemical stain was 55.6%, which increased to 88.9% in 5-month diabetic rats.ConclusionsThe occurrence of the ultrastructural changes in STZ rats with diabetic retinopathy is earlier than that of the expression of bFGF.(Chin J Ocul Fundus Dis, 2003,19:348-351)
Objective To observe whether theograde axial flow of retinal ganglion cells (RGC) in diabetic rats at the early stage was damaged. Methods Diabetic model was induced by streptozotocin in 6 adult male Sprague-Dawley (SD)rats. Fluorogold (FG) was injected to the superior colliculi 4 weeks later.Streched preparation of retina was made 12 and 72 hours after the injection, and was stained after photographed by fluorescent microscope. The proportion of RGC with different sizes labeled by FG was calculated. Other 6 normal adult male SD rats were in the control group. Results Twelve hours after injection with FG, there was no difference of the total number of RGC in experimental and control group, but the ratio of small RGC was lower in experimental group than that in the control group; 72 hours after injection with FG, The number of RGC, especially the small RGC, decreased obviously in experimental group compared with the control group. Conclusion The speed of the retrograde axial flow of RGC in diabetic rats at the early stage is affected, and the small RGC are damageable. (Chin J Ocul Fundus Dis, 2006, 22: 4-6)
Objective To observe the serum betatrophin levels in patients with type 2 diabetes mellitus (T2DM) and to explore the role of betatrophin in the pathogenesis of diabetic retinopathy (DR). Methods A total of 59 patients with T2DM (DM group) and 14 healthy controls (NC group) were enrolled in the study. Vision, slit lamp microscope, indirect ophthalmoscope, fluorescein fundus angiography were performed on all the subjects. According to the results of the examination combined with the international DR clinical staging criteria, the patients were divided into no DR (Non-DR) group, non-proliferative DR (NPDR) group, and proliferative DR (PDR) group, with 30, 20 and 9 patients in each, respectively. The fasting blood glucose (FPG), insulin (FIN), C-peptide, glycated hemoglobin (HbA1c), total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL-C), low-density lipid Protein (LDL-C) levels were detected. The level of betatrophin in serum was determined by enzyme-linked immunosorbent assay. The correlation between betatrophin and other indicators was analyzed by Spearman correlation. The influencing factors of PDR were analyzed by logistic regression. Results Compared with subjects in the NC group, the level of FPG (F=-4.316, P＜0.001), FIN (F=2.142, P=0.001), HbA1c (F=-5.726, P＜0.001), TC (t=3.609, P=0.010), LDL-C (t=0.000, P=0.003), and betatrophin (F=-2.263, P=0.024) were significantly increased and HDL-C level (F=-3.924, P＜0.001) was decreases in the DM group. The difference of TG level between two groups was not statistically significant (F= -1.422, P=0.155). Compared with the Non-DR group and the NPDR group, the serum C-peptide (F=7.818, P=0.020) and betatrophin levels (F=12.141, P=0.002) were significantly increased in the PDR group. Spearman correlation analysis showed that the levels of betatrophin in the DM group was positively correlated to TC (r=0.304, P=0.019). The serum levels of betatrophin was positively correlated to body mass index in the Non-DR group (r=0.513, P=0.004). Furthermore, in the PDR group, a significant positive correlation was observed between the serum betatrophin levels and diastolic blood pressure (r=0.685, P=0.042). Logistic regression analysis showed that the duration of diabetes, serum C-peptide and betatrophin levels were risk factors for PDR. After controlling for the duration and serum C-peptide, the PDR risk for betatrophin levels great than or equal to 1.0 ng/ml was 12 times as much as betatrophin levels less than 1.0 ng/ml in T2DM patients. Conclusions The serum betatrophin content of patients with T2DM is abnormal. Betatrophin may be involved in the occurrence and development of PDR.
Objective To determine the clinical efficacy of probucol in patients with diabetic macular edema (DME) and elevated serum lipids after focal/grid laser photocoagulation. Methods A prospective randomized controlled study included 48 type 2 diabetic patients with DME and dyslipidemia which were randomly divided into three groups. For patients with bilateral disease only the more severe eye was included. All patients were subjected to strict metabolic and blood pressure control during enrollment. All cases received macular laser photocoagulation. Besides, sixteen patients in group A were treated with probucol, 16 members in group B with atorvastatin and 16 members in group C were not treated with any lipid-lowering therapy for about three months. The outcome measurements were status of macular edema and hard exudates, visual acuity, foveal thickness, serum lipids and urine 8-hydroxydeoxyguanosine (8-OHdG) during the three months. Results The study included 20 men and 28 women with noninsulin dependent diabetes mellitus who could achieve good metabolic and blood pressure control within three months of inclusion in the study. Thirteen of 16 patients in group A, twelve of 16 patients in group B and five of 16 patients in group C showed reduction in hard exudates. Regression of macular edema was seen in twelve patients in group A, 11 in group B and eight in group C (χ2=2.368,P＞0.05). The difference of foveal thickness in group A, B and C was statistically significant (t=4.929, 4.669; P=0.000). Nine patients in group A, eight in group B and six in group C showed improving of visual acuity (χ2=1.169,P＞0.05). Three months after treatment, triglycerides (TG) (t=7.954, 6.832; P＜0.05), total cholesterol (TC) (t=6.643, 5.368; P＜0.05) and low-density lipoprotein cholesterol (LDLC) (t=3.279, 3.835; P＜0.05) decreased in group A and group B but not in group C, and high-density lipoprotein cholesterol showed no significant difference in the three groups. 8-OHdG decreased gradually during the first and third month in group A and group B but not in group C. In the first month post treatment, 8-OHdG showed no difference between group A and group B. In the third month, the 8-OHdG was lower in group A than group B, and the difference was statistically significant (t=2.947,P＜0.05). ConclusionsIn type 2 diabetes patients with DME and dyslipidemia, oral probucol can reduce the severity of hard exudates and macular edema, improve the visual acuity, and inhibit the levels of TG, TC, LDLC and 5-OHdG. The effect of probucol was similar to atorvastatin. Probucol could be an adjunct treatment of those patients.
The islet transplantation site can be divided into two categories: orthotopic islet transplantation and ectopic islet transplantation. Orthotopic islet transplantation refers to that the insulin secreted and released from the transplanted islet will be metabolized into the liver through the hepatic portal vein system, which does not change the original insulin metabolic pathway, including the portal vein of the liver, the greater omentum. The insulin secreted by the ectopic islet transplantation changes the original metabolic pathway of insulin. The ideal islet transplantation site generally has the following characteristics: high success rate transplantation, high long-term survival rate of graft, simple operation, less trauma, less complications, low risk, easy to repeat detection and so on. This article provides a review of the current research status of each islet transplantation site, in order to provide reference for future related research.
Purpose To study changes of cell cycle of vascular endothelial cell in non-proliferative diabetic retinopathy. Methods Alloxan induced Wistar-rats were employed and immunohistochemistry,Western blotting methods were used. Results The vascular endothelial cells of retinas of 8～20 weeks diabetic rats were observe to be cyclinD1,cyclinD3,cyclinB1,p21 and p27 positive stained with light and electronmicroscopies.CyclinE immuno-stained vascular endothelial cells was observed occasionally.Meanwhile,the evidences of morphologic changes of the vascular en dothelial cells were proved:less plasma,thinner cell,more bubble organelles than those of controls.But,the ultra-structures of pericytes and other type of retinal cells did not change and they also immunostain negative.Komas blue and Western blotting methods also proved that the vascular endothelial cells of retina of 20th week diabetic rats expressed cyclinD1,cyclinB1,p21 and p27 protein. Conclusion Glucose induced retinal vascular endothelial cells of 8～20th weeks diabetic rats enter cell cycle and were arrested at G1/S restriction point.This study also suggested that retinal vascular endothelial cells may possess the ability to resist glucose damage and mechanism of selfstability during very early stage of diabetes. (Chin J Ocul Fundus Dis,2000,16:173-176)
Objective To explore the related risk factors for diabetic retinopathy (DR) in type 2 diabetes. Methods The clinical data of 412 type 2 diabetes patients, diagnosed between 2003 and 2010, were analyzed retrospectively. The diagnosis of DR and proliferative diabetic retinopathy (PDR) was confirmed by ophthalmoloscopy and fundus fluorescein angiography. Glycated hemoglobin A1c, glucose, insulin, and Cpeptide of fasting plasma, and 1, 2 and 3 hours postprandial plasma were measured. According to the abovementioned data, get the fluctuation of glucose, insulin and C-peptide of 1, 2 and 3 hour postprandial plasma. Results The morbidity of DR and PDR increased following the longer disease duration. Age, diabetic duration，body mass index (BMI), hypertension grade, HbA1C, fasting plasma insulin and C-peptide, 2 and 3 hours postprandial plasma glucose, 1 and 2 hours postprandial plasma insulin, 1, 2 and 3 hour postprandial plasma C-peptide, 1, 2 and 3 hours postprandial plasma glucose, insulin and C-peptide fluctuation are different statistically among non-DR group, non-PDR group and PDR group (P＜0.05). 3 hours postprandial plasma glucose and fasting plasma insulin were risk factors of DR (P＜0.05). Conclusions Postprandial plasma glucose and fasting plasma insulin were risk factors of DR. Nevertheless, postprandial insulin, fasting and postprandial C-peptide, postprandial plasma glucose, insulin and C-peptide fluctuation were useful for DR diagnosis.