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 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 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.
Objective?To explore the glucometabolic state of angiographically documented inpatients with coronary artery disease (CAD) but without diagnosed diabetes mellitus (DM). Methods?The study recruited 449 patients, who were performed a coronary angiography as well as an oral glucose tolerance test (OGTT) when admitted in the cardiovascular medical ward in our hospital from January 2007 to May 2009. According to the results of coronary angiography, the patients were divided into a coronary artery disease (CAD) group and a non-coronary artery disease (non-CAD) group, and abnormal glucose metabolism (AGM) status was compared between the two groups. Results?The random plasma glucose (RPG) and fasting plasma glucose (FPG) had no significant differences (P values were 0.249 and 0.444, respectively) in the two groups, while the OGTT 2-hour plasma glucose (2hPG) was much higher in the CAD group, with a significant difference (Plt;0.001) compared with the non-CAD group. The CAD group had a prevalence of AGM up to 74.0%, of which 32.1% were newly diagnosed DM patients, and 39.0% were impaired glucose tolerance (IGT) patients, much higher than that in the non-CAD group, respectively, there being a significant difference (P=0.006). Logistic regression analyses revealed that the risk of IGT and newly diagnosed DM was 1.6 times (OR=1.603, 95% CI 1.023 to 2.512, P=0.04) and 2.3 times (OR=2.292, 95% CI 1.391 to 3.777, P=0.001) as much as that in non-CAD patients, respectively; when adjusted for the factors such as hypertension, dyslipidemia, BMI, hs-CRP, and other factors, CAD patients still had a higher risk of newly diagnosed DM (OR=1.852, 95%CI 1.064 to 3.223, P=0.029), compared with the non-CAD patients. Conclusion?AGM is common in the admitted patients with CAD but undiagnosed diabetes, most of whom need an OGTT to be diagnosed timely and accurately. OGTT should be considered to be a routine inspection item to diagnose AGM in the inpatients with CAD; if possible, all hospitalized patients with cardiovascular disease should be performed an OGTT routinely.
Objective To analyze the expression of apoptosis-related genes of retinal blood vessel in early diabetic rats by gene chip technology. Methods To make diabetic rat model by intraperitoneal injection of streptozotocin (STZ). On the 6th week after blood pressure increased, 10 rats were executed in Diabetic group and normal control group respectively. 20 retinal blood vessels were extracted and the RNA was isolated. The probe was made of alpha;-32 P-deoxyadenosine triphosphate (dATP)-labeled sample which hybridized 1176 nylon chips, and then analyzed by software. Three different expression genes were selected to verify by reverse transcription polymerase chain reaction (RT-PCR). Results On the 6th week, 136 （11.5％） genes were differentially expressed ［up-regulated genes were 90(7.6%), down-regulated genes were 46(3.9%)］in diabetic group. These genes involved into different groups according to their function. Especially in 72 apoptosis-related genes, 15 genes were differentially expressed. The up-regulated genes were some TNF receptor family members such as TNFRSF12, TRAIL, TNFRSF9, FADD；Bcl-2 family members such as bcl-w, bax, bak1 and AKT. The down-regulated genes were FAF1 which related to fas. Conclusions The expression of retinal vascular gene in early diabetic rats has been changed complicatedly. In particular, the multiple apoptosis-related genes have been changed in early diabetic, and most of them are at the upstream of apoptosis pathway. These findings indicate that the development of diabetic retinopathy is associated with multiple signaling pathways leading to apoptosis, while the alterations on the level of molecular biochemistry are still limited in apoptosis induction period. （Chin J Ocul Fundus Dis，2008，24：244-248）
Diabetic kidney disease (DKD) is a major complication of diabetes mellitus. One third of patients with advanced diabetes mellitus can develop to uremia, which seriously endangers people’s health. In recent years, with the improvement of people’s living standards and the increasing incidence of diabetes, it has become the main cause of end stage renal disease in China. Over the past two decades, the understanding of diagnosis and treatment of DKD has been improved, such as putting forward the new concept of normoalbuminuric DKD and developing a variety of new anti-diabetic drugs. However, at present, the basic strategies of DKD treatment are still lifestyle modification, glucose control, blood pressure control and lipid control.
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)
ObjectiveTo investigate the effects of interferon gene stimulating protein (STING) inhibitor (C176) on human retinal microvascular endothelial cells (hRMEC) under oxidative stress. MethodsAn animal experimental study. In vivo experiment: 48 healthy male C57BL/6J mice were randomly divided into wild type mice group (WT group) and diabetes (DM) group, with 24 mice in each group. DM mice were induced by streptozotocin to establish DM model. After successful modeling, DM group was divided into DM+dimethyl sulfoxide (DMSO) group and DM+C176 group, with 12 mice in each group. The mice in the DM+DMSO group were intraperitoneally injected with DMSO at the dose of 50 mg/kg. Mice in DM+C176 group were intraperitoneally injected with STING inhibitor C176 750 nmol at the dose of 50 mg/kg. Four weeks after modeling, immunohistochemical staining, Western blot and real-time fluorescence quantitative polymerase chain reaction were used to detect the expression of STING in the retina of WT and DM mice. The leukocyte adhesion test was used to detect the number of leukocytes adhering to hRMEC in mice with WT, DM+DMSO and DM+C176 groups. In vitro experiment: hRMEC was randomly divided into conventional culture cell group (N group), dimethyl sulfoxide (DMSO) group (with DMSO intervention) and C176 group (with C176 intervention). The cells were induced by 150 μg/ml glycation end products for each group. In vitro leukocyte adhesion test combined with 4', 6-diamino-2-phenylindole staining was used to detect the number of leukocytes adhering to hRMEC. The adherent leukocytes were quantitatively analyzed by flow cytometry; H2DCFDA/reactive oxygen species (ROS) fluorescence probe was used to detect ROS expression in cells; Seahorse XFe96 cell energy metabolism analyzer was used to measure the level of intracellular glycolysis. t-test was used to compare the two groups; single factor analysis of variance was used to compare the three groups. ResultsIn vivo experiment: compared with WT group, the expression level of STING (t=73.248) and the relative expression amount of mRNA (t=67.385) in the retina of DM group mice increased significantly (P＜0.05). Compared with WT group, the number of leukocytes adhering to the retinal vessels of mice in DM+DMSO group was significantly increased, while that in DM+C176 group was significantly decreased (F=84.352, P＜0.01). In vitro: compared with N group and DMSO group, the number of leukocyte adhesion on hRMEC in C176 group decreased significantly (F=35.251, P＜0.01). Compared with N group, the number of leukocytes adhering to hRMEC in DMSO group and C176 group decreased significantly (F=26.374, P＜0.01). The ROS level in hRMEC in C176 group was significantly lower than that in N group and C176 group (F=41.362, P＜0.01). Compared with N group and DMSO group, the glycolysis level of hRMEC in C176 group was significantly reduced, with a statistically significant difference (F=68.741, P＜0.01). ConclusionInhibiting the expression of STING in retinal vascular endothelial cells can improve the progress of DM by inhibiting leukocyte adhesion, ROS production and glycolysis level.
Objective To explore the effects of CO2 pneumoperitoneum on pancreatic function in diabetic rabbits. Methods Forty-eight rabbits were divided into 4 groups: control group (the group of N0, n=4), the group of T0 (n=4), the group of T10 (n=20), and the group of T15 (n=20). The animal used in the groups of T0, T10 and T15 was diabetic rabbit, and the pressures of pneumoperitoneum of the three groups were 0 mm Hg, 10 mm Hg and 15 mm Hg respectively.The model of diabetic rabbits were made through intrvenous administration of Allxon. Arterial blood samples were collected before the onset of CO2 pneumoperitoneum, 0, 2, 6, 12 hours after deflation for measuring blood glucose, amylase, insulin and C-peptid. Then the rabbits were sacrificed and their pancreases were removed for measuring SOD activity and MDA content. Results After abdominal deflation, the blood glucose, amylase, insulin, C-peptid, MDA content were significantly increased （P＜0.05), and SOD activity was significantly decreased（P＜0.05). Twelve hours after abdominal deflation, the levels of blood glucose, amylase, insulin, C-peptid, MDA content returned to those before pneumoperitoneum was established in group T10. But, those in group T15 were higher （P＜0.05) than the levels before insufflation. The SOD activities in both group T10 and group T15 twelve hours after abdominal deflation were significantly different （P＜0.05） from those before pneumoperitoneum was established. There were statistically significant differences （P＜0.05) between group T10 and T15 in amylase, C-peptid, MDA content and SOD activity. Conclusion CO2 pneumoperitoneum has an certain adverse influence on pancreatic function of the diabetic rabbits. The degree of injury is correlated with the pressure of pneumoperitoneum. Pancreatic function may returned to preoperative level soon after abdominal deflation in group T10, but did not return in group T15.
ObjectiveTo observe the features of the full field electroretinogram (FF-ERG) in type 1 diabetes (T1D) children without diabetic retinopathy (DR). MethodsRetrospective case study. Forty-one T1D children and 25 age-matched normal controls underwent a complete ophthalmic examination, including best-corrected visual acuity, refraction, intraocular pressure, slit lamp, fundus photography, indirect ophthalmoscopy, and spectral domain optical coherence tomography to exclude DR. All FF-ERG tests were performed by an experienced technician. The ERG series includes six protocols: dark-adapted 0.01 ERG (r-b 0.01); dark-adapted 3 ERG (mix-a 3.0, mix-b 3.0); dark-adapted 10 ERG (mix-a 10.0, mix-b 10.0); dark-adapted oscillatory potentials (OPS); light-adapted 3 ERG (c-a 3.0, c-b 3.0); light-adapted 30 Hz flicker (30 Hz FP) ERG. To compare the amplitudes and implicit times of the FF-ERG between the T1D and control group children. ResultsCompared with the control subjects, the FF-ERG amplitudes decreased and the implicit times increased in T1D. Except for r-b 0.01 (t=-0.228, P > 0.05), the amplitudes of other FF-ERGs were all significantly attenuated (t=-1.664, -3.645, -4.324, -6.123, -5.846, -12.9, -14.4, -5.23; P < 0.05) in T1D children. The implicit times of mix-b 3.0, mix-b 10.0, c-b 3.0 and OP2 significantly increased (t=5.242, 2.879, 5.378, 3.506; P < 0.05). The implicit times of r-b 0.01, mix-a 3.0, mix-a 10.0, c-a 3.0 and 30Hz FP changes were not significantly (t=2.331, 1.677, 0.557, 0.84, 0.064; P > 0.05). ConclusionThe FF-ERG amplitudes decreased and implicit times increased in T1D children compared with the control normal subjects.