Objective o observe the expression of Notch1 and Delta-like ligand 4 (Dll4) on the fibrovascular membranes in proliferative diabetic retinopathy (PDR), and investigate its relationship with vascular endothelial growth factor receptor 2 (VEGFR2). Methods Fifty-seven PDR patients (60 eyes) who underwent vitrectomy were enrolled in this study. The PDR patients were divided into non-injection group (30 patients, 32 eyes) and injection group (27 patients, 28 eyes). The eyes in injection group received intravitreal injection with ranibizumab at 2 to 7 days before surgery. The preretinal fibrovascular membranes were obtained from the PDR patients during vitrectomy. Eighteen epiretinal membranes were obtained from the non-diabetic patients was served as controls. The real-time polymerase chain reaction (RT-PCR) and immunohistochemical methods were used to detecting the expression of Notch1, Dll4 and VEGFR2. In the meantime, the numbers of the nucleus of vascular endothelial cells in the membranes stained with hematoxylin were counted. Results The immunohistochemical staining revealed that there were positive expression of Notch1, Dll4 and VEGFR2 in all PDR membranes, regardless of the injection of the ranibizumab. The levels of Notch1, Dll4 and VEGFR2 protein in non-injection group were higher than those of injection group (t=3.45, 6.01, 4.08;P=0.030, 0.008, 0.023). In injection group, the number of endothelial cells in the membranes reduced (17.17±2.48) compared with that of the non-injection group (41.50±5.57). There was significant difference in the number of endothelial cells in the membranes between the two groups (t=9.58,P＜0.05). RT-PCR showed that the differences of the mRNA expression of Notch1, Dll4 and VEGFR2 were all statistically significant among the PDR group and control group (H=12.50, 12.50, 12.02;P＜0.05).The expression of Notch1, Dll4 and VEGFR2 in the PDR membranes was higher than that of epiretinal membranes from non-diabetic patients. In the PDR group, the expression of Notch1, Dll4 and VEGFR2 of non-injection group was higher than that of injection group. Spearman correlation analysis showed that the expression of mRNA between VEGFR2 and Dll4 (r=0.83), VEGFR2 and Notch1 (r=0.81), Notch1 and Dll4 (r=0.87) were all significantly correlated (P＜0.05). Conclusions The expression of Notch1 and Dll4 in the PDR membranes are higher than that of the control group, and it is positively correlated with the expression of the VEGFR2. Notch1 and Dll4 play a regulatory rule in the neovascularization in PDR, the acting way may be correlated with VEGFR2.
Objective To investigate auto-cortex of crystalline lens-induced neovascular epiretinal membrane（NVERM）by micro-injuring posterior c apsule of crystalline lens. Methods twenty four C57BL/6 mouse between 4-6 weeks were selected, and divided into two groups randomly: auto-cortex of crystalline group and the control group. The auto-cortex of crystalline group was treated by penetrating the posterior capsule of lens and washing out the lens cortex into the mouse vitreous using PBS (phosphate buffered solution), while the control group were injected PBS into vitreous merely. Clinical change s were followed by slit-lamp examination and photograph. The eye balls were enu cleated at the day of 3, 7, 14 and 28 after operation. Both HE and immunohistoch emistry were used to detect the pathological changes. Results postoperative one to three days, 11 of 12 mouse in autocortex of crystalline g roup, lens appear to alba turbid at different levels one after another, and then develop into highdensity chinaware white. Postoperative (po) three days, HE s taining shows cortex of lens debris transmigrated in vitreous cavity, and some o f which approached to internal limiting membrane and lead it to rough and discon tinue; Po7-14 days, the capillary in retina expanded, migrated and broke though t internal limiting membrane which got to the pro retina and became the new ves sels. And typical NVERM were observed. Po28 days, some vascularslike structure formed in vitreous cavity. None of mouse in control group developed NVERM. Conclusion Auto-cortex of crystalline lens can induced neovascular epiretinal membrane in C57BL/6 mouse. (Chin J Ocul Fundus Dis,2008,24:118-121)
Objective To investigate the viability and the characters of proliferation and differentiation of retinal stem cells (RSCs) after cryopreservation and anabiosis. Methods The RSCs of a Long Evans rat with the embryonic age of 17 days were separated and cultured in vitro. The third-passage RSCs in the cryopreservation liquid consisted of 80% Dulbecco modified Eagle medium (DMEM)/F12，10% bovine serum albumin (BSA)，10% dimethylsulfoxide (DMSO)，and basic fibroblast growth factor (bFGF) (20 ng/ml), were stored in liquid nitrogen. After 1, 2, 4, 8, 12, and 16 weeks of freezing period, these cells were thawed. The livability of the cells was counted and the differentiation was induced while the proliferation and characters of differentiation were detected by immunofluorescence. Results The effects of different durations of cryopreservation on the livability of RSCs did not differs much (Pgt;0.05). These cells were reculturd well and presented specific marker of RSCs. In addition, they also could be induced and differentiated into several types of retinal cells. Conclusion Cryopreservation and anabiosis of RSCs does not affect the cellular intrinsic characters of proliferation and differentiation. (Chin J Ocul Fundus Dis, 2007, 23: 94-97)
Objective To explore the changes in the serum Cu2+and the indexes for the liver pathology and biochemistry before and after the copper needle retained in the central veins of the rabbit ears. Methods Fortynine New Zealandrabbits were randomly divided into 5 groups: Group A (copper needles retained for 1 d), Group B (3 d), Group C (5 d), Group D (7 d), and Group E (the control group, without any copper needles retained). In each experimental group, there were 10 rabbits, and in the control group there were 9 rabbits. The rabbits in each group were arranged to have their venous blood drawn for determination of theCu2+concentration, and for observation on the changes in the liver biochemicalindexes for 5 times before and after the copper needles were retained in the central veins of the rabbit ears. At the same time, a piece of the liver tissue ineach rabbit was taken for examination of the pathological changes. All the liver samples were given the basic pathological examination; if the liver sample hadsome extraordinary pathological features, the specific pathological examinationwould be given, even using the transmission electron microscope. Results After the copper needles were retained in the central veins of the rabbit ears, the Cu2+concentration increased with the passing time. The concentrations in the groups were 1.40±0.49 μg/ml in Group A, 1.45±0.53 μg/ml in Group B, 2.01±0.40 μg/ml in Group C, 2.38±0.83 μg/ml in Group D, and 1.34±0.45 μg/mlin Group E, respectively. There was a statistically significant difference between Group D and Group E(P＜0.05). There were no significant changes in the ALT and AST levels when compared with those before the copper needles were retained(P＞0.05); however, there was a considerably positive correlation on 1st day (r=0.686, P＜0.05), 5th day (r=0.712, P＜0.05), and 7th day (r=0.768, P＜0.01) when compared with those after the copper needles were retained. The histological examination showed that aseptic inflammation subsided with the time in part of the liver. The Masson staining and the Ag staining showed that there were no obvious changes in the hepatic lobules, with no fibrosis of the liver tissues found under light microscope. Conclusion There are no obvious toxic and side effects on the rabbit liver after the copper needles are retained in its central veins.
Objectivｅ To investigate the histopathologic charact eristic of the vitreous herniation out of sclerotomy site during vitrectomy. Methods Twenty specimens of tissues herniated at vitrectomy site were collected. The paraffin sections or fresh smears were stained with hematoxylineosin and examined under light microscope. The specimens were collected from the affected eyes with rhegmatogenous retinal detachment (9 cases), traumatic retinal detachment (1 case), miscellaneous vitreous hemorrhage (6 cases) and intraocular foreign body (4 cases). Results The herniated tissues were found to be retina in 4 cases, ciliary tissue in 1 case, retina and ciliary tissue in 1 case, uvea in 1 case, and hyaloid tissue in 13 cases. Conclusion There were not only vitreous, ciliary epithelial cells and pigment containe depithelia, but also ciliary body, retina and uvea in the prolapsed tissues of sclerotomy site, which might be related to the occurence of some clinical complications. (Chin J Ocul Fundus Dis,2001,17:99-101)
ObjectiveTo analyze the relationship between maximum standardized uptake value (SUVmax) of primary tumor detected by 18F-FDG positron emission tomography/computed tomography (PET/CT) and clinicopathologic factors in stageⅠnon-small cell lung cancer (NSCLC), and investigate the prognostic value of PET/CT on pathological feature. MethodsWe retrospectively analyzed clinical data of 182 patients with stageⅠNSCLC who underwent 18F-FDG PET/CT scan before lobectomy or segmentectomy in China-Japan Friendship Hospital from April 2013 to June 2014. There were 121 male and 61 female patients with their ages of 34-85 (68.1±9.8) years. Clinicopathologic factors including sex, age, smoking history, histology, TNM stage, T stage, tumor size, lymphatic vessel invasion, blood vessel invasion (BVI) and visceral pleural invasion were evaluated to identify the independent factors affecting SUVmax by univariate and multivariate regression analysis. The diagnostic efficiency and best cut-off point of SUVmax were calculated by the receiver operating characteristic curve. ResultsThe univariate analysis identified that sex (P=0.015), smoking history (P=0.001), histology (P < 0.001), TNM stage (P=0.004), T stage (P=0.001), tumor size (P < 0.001), BVI (P=0.001) were factors affecting SUVmax. Only histology (P=0.001), tumor size (P=0.006), BVI (P=0.009) were found to be significant independent factors according to multivariate regression analysis. The SUVmax of primary tumor was a predictor for BVI with the highest diagnostic accuracy at a cut-off value of 4.85, the sensitivity and specificity were 65.5% and 71.7%. ConclusionThe SUVmax is correlated with histology, tumor size and BVI in stageⅠNSCLC, higher in patients with non-adenocarcinoma, lager tumor and positive BVI. Furthermore, the probability of BVI could be predicted by SUVmax of the primary tumor.
ObjectiveTo observe the changes of glaucoma optic nerve head (ONH) parameters and macular ganglion cell complex (GCC) structure in preperimetric glaucoma (PPG) patients. Methods Eighteen PPG patients (18 eyes, PPG group), 22 primary open-angle glaucoma (POAG) patients (22 eyes, POAG group), and 20 patients (20 eyes) with physiologic large optic cup (physiological big optic cup group) were included in this study. Seventeen healthy volunteers (17 eyes) were the normal control. The optic nerve head and macular was scanned by fourier-domain optic coherence tomography (FD-OCT) for all subjects. The following 15 parameters, including nerve fiber layer thickness (RNFL), the optic disk rim volume (RV), optic nerve head volume (NHV), optic disc area (ODA), rim area (RA), cup volume (CV), cup/disc area ratio (CDAR), vertical cup/disc ratio (VCDR), horizontal cup/disc ratio (HCDR) and optic cup area (CA), macular GCC, superior GCC, inferior GCC thickness, focal loss of volume (FLV) and global loss of volume (GLV), were measured at 10 different quadrants. The relationship between macular GCC thickness or optic disc RNFL thickness and RA was analyzed by simple linear regression analysis. ResultsThe RNFL thickness of PPG patients was (99.29±19.93) μm (superior quadrant), (97.29±22.86) μm (inferior), (114.61±15.64) μm (superior temporal, ST), (119.22±26.19) μm (inferior temporal, IT), (116.11±39.32) μm (superior nasal, SN), (111.33±37.65) μm (inferior nasal, IN), (77.56±17.22) μm (temporal upper, TU), (76.78±10.34) μm (temporal lower, TL), (88.94± 42.54) μm (nasal upper, NU), and (82.33±43.83) μm (nasal lower, NL) respectively, which was thinner than normal control group and physiologic large cup group, but thicker than POAG patients. Compared to normal controls and physiologic large cup patients, PPG patients also had 4 parameters reduced (RV, NHV, ODA and RA), and 5 parameters increased (CV, CDAR, VCDR, HCDR and CA), the differences are statistically significant (P < 0.05). However, these parameters were similar to POAG patients (P > 0.05). For macular GCC parameters, PPG patients also had 3 parameters reduced (average GCC, superior and inferior GCC thickness), and 2 parameters increased (GLV and FLV) compared to normal control group and physiologic large cup patients (P < 0.05). However, these parameters were similar to POAG patients (P > 0.05).Simple linear regression analysis showed that, with the GCC macular thinning, reducing the number of ganglion cells reduced, optic disc RNFL thickness became thinner (regression coefficient=1.25, P=0.00) and RV reduced (regression coefficient=0.037, P=0.00). ConclusionsPPG patients and normal control had a similar distribution of optic disc RNFL. Five parameters (RV, NHV, ODA, RA, macular GCC thickness) were less than normal control and physiological big optic cup group, but had no significant differences compared with POAG group.