Objective To observe the expression of matrix metalloproteinase（MMP-2, MMP-9 and vascular endothelial growth factor (VEGF) in retinoblastoma (RB) and its relationship with the differentiation and optic nerve infiltration of RB.Methods Forty paraffin specimens of pathological confirmed RB were studied. They were divided into differentiated group (15 cases) and undifferentiated group (25 cases) , optic nerve infiltration group（13 cases） and without optic nerve infiltration group（27 cases）. The expression of MMP-2, MMP-9 and VEGF were detected by immunohistochemistry, their relationships with the differentiation and optic nerve infiltration were also analyzed.Results The positive rate of MMP-2, MMP-9 and VEGF expression in 40 RB cases were 52.5％，57.5％ and 72.5% respectively．The expression of MMP-2, MMP-9 and VEGF in the undifferentiated group were significantly higher than those in the differentiated group (chi;2=9.037, 9.253, 8.095; P＜0.05). The expression of MMP-2, MMP-9 and VEGF in RB with optic nerve infiltration group were significantly higher than those in RB without optic nerve infiltration group (chi;2=11.045,10.243, 8.956；P＜0.05). The expression of MMP-2，MMP-9 had a positive correlation with the expression of VEGF in RB (r=0.126,0.314；P＜0.05). Conclusions MMP-2, MMP-9 and VEGF expressed in RB tumor tissues. The expression of MMP-2, MMP-9 has a positive correlation with the expression of VEGF. The levels of MMP-2, MMP-9 and VEGF expression are related to optic nerve infiltration of RB cells.
Objective To expolre the factors which affect the size of diabetic,macular fobeal avascss scular zone(FAZ). Methods Making ten years of duration of diabetes a limit,79 nonproliferative and early proliferative diabetic patients were divided into 2 groups.Diabetic retinopathy severity level was diveided into 4 stages,and the macular edema was subdivided into focal、diffuse and cystoid according to fluorescein leakage of foveomacular region.All patients were measured FAZ with Heidelberg scanning laser fluoresceion angiography system and then compaired the size of FAZ of patients with different duration of diabetes、diabetic retinopathy severity level and macular edema status.The results were performed analysis of variance and t test. ResultsThe study shown the size of FAZ was not directly related to the duration of diabetes(t=1.3854,Pgt;0.1);There were significant differences about the size of FAZ of patients with different diabetic retinopathy severity level(F=7.6251,P＜0.01)and macular edema status(F=5.4369,P＜0.01)． Conclusion The size of FAZ was significantly increased in diabetic patients.It was enlarged with the development of diabetic retinopathy severity level,but it was not related to duration of diabetes. (Chin J Ocul Fundus Dis,2000,16:155-156)
ObjectiveTo explore the risk factors, pathophysiological mechanism, pathogenic bacteria distribution, diagnosis, and treatment of postoperative intra-abdominal infection, and to provide a theoretical basis for further understanding the mechanism and treatment of postoperative intra-abdominal infection.MethodThe related literatures in PubMed, CNKI, WanFang, and other databases were searched to summarize the research progress of postoperative intra-abdominal infection.ResultsPostoperative intra-abdominal infection was associated with a variety of risk factors, and timely identification and control were conducive to the prevention of intra-abdominal infection. Postoperative intra-abdominal infection had a complex pathophysiological mechanism, mainly involving changes in the immune system, which provided a target for immunotherapy. Pathogenic bacteria were widely distributed in postoperative intra-abdominal infection, and the problem of drug resistance was also a big problem nowadays. In the treatment of postoperative intra-abdominal infection, comprehensive treatment measures should be taken to control the infection, in which the control of the source of infection was the basis and played a key role.ConclusionsThe treatment of postoperative intra-abdominal infection needs to be more individualized and refined, and comprehensive treatment measures such as controlling the source of infection, nutrition therapy, organ function support, and so on, should be taken. Immunotherapy is a new potential treatment measure.
Objective To explore and evaluate the protective effects of soluble fms-like tyrosine kinase recptor-1(sFlt-1) gene 2-3 and 2-4 transcellular region on retinal vascular leakage and phosphatidyl inositol-3 kinase (PI3K)/Akt pathway under hypoxia and/or hyperglycemia. Methods The plasmids pcDNA3.1-EGFP/sFlt-1(2-3) and pcDNA3.1-EGFP/sFlt-1(2-4) were constructed.. Two of these plasmids with carboxy methylation glucan (CMD) magnetic particles were transfected into human umbilical vein endothelial cells (HUVEC), which were cultured under hypoxia and/or hyperglycemia. The blood retinal barrier was evaluated by Evans blue permeation (EBP). Then the expression of p-Akt mRNA and protein were detected by reverse transcriptionpolymerase chain reaction (RT-PCR) and Western blot separately. Results In the diabetic rabbits, The blood retinal barrier breakdown was detected by the retinal vascular leakage of EBP. The expression of p-Akt mRNA and protein in hypoxia and hyperglycemia groups were also obviously increased. These changes were largely prevented by transfection the plasmids pcDNA3.1-EGFP/sFlt-1(2-3) and pcDNA3.1-EGFP/sFlt-1(2-4) (P＜0.01 in both groups). Conclusion Both sFlt-1(2-3) and sFlt-1(2-4) can make the retinal blood vessels less leaky and may be beneficial in preventing vision loss from diabetic retinopathy.
Objective To investigate the possible mechanism of arsenic trioxide (As2O3) inducing P16 gene demethylation and transcription regulation in the retinoblastoma (RB) Cell Line Y79． Methods The induced growth inhibition of Y79 cell was assayed by MTT; The DNA content of Y79 cell was analyzed by flow cytometry after being exposed to As2O3; the methylation status of the P16 gene in Y79 cell line before and after treatment with As2O3 was detected by the nestedmethylation specific PCR and DNA sequencing; the mRNA of P16,DNA methyltransferases (DNMT3A and 3B)gene were determined by RT-PCR． Results As2O3 was able to inhibit the growth of Y79 cell and increase the cell number in G0-G1 phase；P16 gene was not expressed in Y79 cell line and As2O3 can induce itrsquo;s mRNA expression；after 48 hour disposal of As2O3，the methylation levelof P16 gene was apparently attenuated in Y79 cell line,the expression of DNMT3A and DNMT3B was obviously down-regulated． Conclusions P16 gene is the hypermethylation in the retinoblastoma cell line Y79, and As2O3 can inhibite the methylation of P16 gene and upregulate the expression of p16 gene mRNA which inhibits the proliferation of Y79 cell by inducing the G0-G1 arrest, by inhibiting the expression of DNA methyltransferases.
Objective To investigate the expression of pigment epitheliumderived factor (PEDF) and vascular endothelial growth factor (VEGF) in choroidal melanoma. Methods The expression of VEGF and PEDF protein in fifty-eight cases of paraffinembeded choroidal melanoma samples was measured by immunohistochemistry, the expression of PEDF mRNA in thirtynine choroidal melanoma samples was assayed by in situ hybridization. Results PEDF protein was detected in 13/58 cases (22.4%) of choroidal melanoma, the positive rate in nonsclerainvasion group (12/38, 31.6%) was higher (Plt;0.05) than that in sclerainvasion group (1/20, 5%). VEGF protein was detected in 43/58 cases (64%) of choroidal melanoma, the positive rate in nonsclerainvasion group (25/38, 65.8%) was lower (Plt;0.05) than that in sclerainvasion group (18/20, 90%). The expression of PEDF mRNA was detected in 18/39(46.2) cases, the positive rate in nonsclerainvasion group was higher (Plt;0.05) than that in sclerainvasion group. Conclusions Imbalanced expression of VEGF and PEDF in choroidal melanoma may play a key role in the angiogenesis, tumor progression and metastasis.
Objective To study the proximal diameter changes of retinal blood vessel following branch retinal vein occlusion (BRVO). Methods Color fundus photographs and fundus fluorescein angiography (FFA) photographs of 48 patients with typical unilateral BRVO were analyzed using IMAGEnet software. The diameter of retinal artery (RAD) and vein (RVD) close to optic disc (within one DD from the optic disc) in four quadrants including the affected quadrant were measured with linear measuring tools.Results The proximal diameter of RAD and RVD in corresponding normal quadrants of the BRVO eye had no significant change comparing with the contralateral eye. The proximal diameter of RAD, but not RVD of the affected quadrant such as superotemporal （t=-2.342, P=0.026）or inferotemporal （t=-3.069, P=0.010）quadrant, increased remarkably. Conclusions In corresponding affected quadrant with BRVO, only RAD close to optic disc increases markedly, RVD has no significant change.
Objective To observe the oxidative damage of mtDNA, apoptosis and expression of adhesion molecules in retinal capillary cells of diabetic rat with different disease courses. Methods One hundred Sprague-Dawley rats were randomly divided into the control group and the experimental group. The rats of experimental group were induced with streptozotocin (STZ) injection creating a diabetic model. Then they were divided into DR1m, DR2m DR3m group according to disease courses. The rats of control group were divided into NR1m, NR2m, NR3m group. Rat retinal capillaries were prepared, and then the contents of undamaged mtDNA were examined by Southern blot combined with Fpg. The expression of cyclooxygenase (COX)-1 encoded by mtDNA and transcription factors A (mtTFA) mRNA were detected by real-time quantitative polymerase chain reaction (RT-PCR). Apoptosis and expression of intercellular adhesion molecule-1 (ICAM-1) were detected by terminal dUT nick endlabeling (TUNEL) immuno-fluorescence and immunohistochemistry respectively. Results The contents of undamaged mtDNA in rats of DR1m, DR2m, DR3m were less than those of NR1m、NR2m、NR3m. The contents of undamaged mtDNA in diabetic rats decreased with the increase of disease courses. In addition, the mRNA levels of COX-1 and mtTFA were downregulated in diabetic rats. The positive cells of TUNEL and ICAM-1TUNEL and ICAM-1 in diabetic rats increased with the increase of disease courses. Conclusion With the increase of disease courses, mtDNA damage and apoptotic cells are increased, while the expression of mRNA encoded by mtDNA and ICAM-1 decreased in retinal capillary cells in diabetic rats.
Objective To observe the changes of tortuosity and bifurcation angle of retinal arteries and veins in each quadrant of the posterior pole in eyes with high myopia.Methods The tortuosity and bifurcation angle of retinal vessels in each quadrant of the posterior pole in 32 patients (52 eyes) with high myopia and 22 healthy people (30 eyes) were observed and compared. The outcomes were analyzed by multivariate analysis of variance. Results The tortuosity of macular vessels and the artery from optic disc in eyes with high myopia was (1.29plusmn;1.10)times;10-4 and (5.39plusmn;1.93)times;10-5 respectively, and in the normal eyes was (4.15plusmn;2.38) times;10-4 and (9.75plusmn;4.99)times;10-5 respectively; there was significant difference between the two groups （t=1.99, 2.00;Plt;0.05）. The bifurcation angle of superior nasal and inferior nasal retinal arteries in eyes with high myopia was（66.17plusmn;14.04）deg; and (61.20plusmn;11.02) deg; respectively, and in the normal eyes was （77.66plusmn;14.12）deg; and (85.86plusmn;16.45) deg; respectively; there was significant difference between the two groups （F=0.77, 0.83; Plt;0.05）. The bifurcation angle of superior temporal and inferior temporal retinal veins in eyes with high myopia was（92.39plusmn;20.36）deg; and (83.56plusmn;23.50) deg; respectively, and in the normal eyes was （79.45plusmn;15.94）deg; and (70.59plusmn;17.27) deg;; there was significant difference between the two groups （F=2.34, 1.83; Plt;0.05）.Conclusions The vessel tortuosity of retinal arteries and the vessels extending from the optic disc to macula is smaller in eyes with high myopia, while the venous tortuosity has no change. The bifurcation angle of retinal arteries in the superior nasal and inferior nasal field was smaller in eyes with high myopia, while the venous tortuosity has no change. The bifurcation angle of retinal veins in the superior temporal and inferior temporal field was larger in eyes with high myopia.
Objective To observe the expression of vascular endothelial growth factor receptor-1 (VEGFR-1) and VEGFR-2 in hypoxic chorioretinal endothelial cells of monkeys (RF/6A), and to evaluate the effect of minocycline. Methods RF/6A was cultured and divided into four groups: control group, hypoxia group, hypoxia and low dose of minocycline group (0.5 mu;mol/L), hypoxia and medium dose of minocycline group (5 mu;mol/L), and hypoxia and high dose of minocycline group (50 mu;mol/L). Real-time reverse transcriptionpolymerase chain reaction (RT-PCR) and immunohistopathological staining were used to measure the mRNA and protein expression of VEGFR-1 and VEGFR-2, respectively. Results RT-PCR showed that the expression of VEGFR-1 mRNA did not vary significantly between groups (F 24 h=0.17,F 48 h=1.53,F72 h=2.04;P＞0.05). Compared with hypoxia group, the expression of VEGFR-2 mRNA in all minocycline treated groups were significantly downregulated (low minocycline, medium minocycline, high minocycline: t=4.69, 20.16, 17.12; P＜0.001). The immunohistopathological study showed the cells with positive staining of VEGFR-1 can be observed in all groups, and the staining was relatively weak and mainly located in cell membrane and cytoplasm. The optical density value analysis showed that the protein expression of VEGFR-1 did not vary significantly between groups at all time points（F 24 h=0.251,F 48 h=0.340,F72 h=0.589;P＞0.05）. The VEGFR-2 positive staining cells were also observed in all groups, and the staining was relatively high. Brown staining particles of VEGFR-2 were observed in the cell membrane with minor staining particles in cytoplasm. The staining density of VEGFR-2 was significantly higher in hypoxia group than control group. Compared with the hypoxia group, the protein expression of VEGFR-2 in minocycline treated groups was significantly lower（F 24 h=19.147,F 48 h=14.893,F72 h==11.984; P＜0.05). Conclusion The expression of VEGFR-2 is upregulated in RF/6A, and minocycline somewhat shows an inhibition effect.