The expression of bcl-2 and p16 in 25 cases of gallbladder mucosa hyperplasia, 40 cases of gallbladder carcinoma and 25 adjacent normal gallbladder mucosa tissue was detected with immunohistochemical staining. Normal gallbladder mucosa was uniformly negative. Bcl-2 and p16 positive rate were significantly higher in hyperplasia and carcinoma than in normal tissue (Ｐ＜0.01). Expression of bcl-2 and p16 got higher with the increasing degree of hyperplasia, p16 was associated with lymph node metastasis and depth of invasion of carcinoma of the gallbladder. The results suggest that bcl-2 and p16 expression play an important role in tumorigenesis, p16 can be used as a prognostic index for gallbladder carcinoma.
Objective To observe the inhibiting effects of alginate sodiumretinoic acid（AGS-RA）microspheres release system on the laser coagulationinduced subretinal proliferation.Methods RA were dissolved by absolute alcohol,then mixed with 1.5% AGS and made into AGSRA microspheres by a microcapsule electrostatic generator. The parameter of laser injury include irradiation time (0.20 s)，spot diameter (200 mu;ｍ) and output power (420 ｍＷ).Thirty pigmented rabbits were randomly divided into 3 groups (laser injury，experimental and control group).After laser coagulation,AGSRA or blank microspheres were immediately injected into the vitrous of experimental and control rabbits respectively.The height,width and area of 6 retinal spots of laser coagulation at each timepoint were analyzed histopathologically with serial retinal sections at 1,2,3,4,and 6 weeks after laser coagulation.Results Histopathological examination showed that there were morphological and distribution changes of retinal cells in all layers, and localized fibroblasts proliferation in the retina after laser injury. The laserinduced responses in experimental group were much milder（P＜0.01）, while the laser injury group and control group have same width（P＞0.05）and height/area of laser spots（P＞0.05）.Conclusion AGSRA release system can alleviate the subretinal proliferate after laser injury.
ObjectiveTo investigate the therapeutic effects of scleral buckling operations on treating longstanding retinal detachment with subretinal proliferation.MethodsThe clinical data of 36 patients (40 eyes) with long-standing retinal detachment with subretinal proliferation who had undergone scleral buckling operation were retrospectively analyzed, and the clinical features, therapeutic methods and curative effects were summarized.ResultsThe diagnosis of long-standing retinal detachment mainly based on the examination of ocular fundus. The features of the affected eyes were: flat retinal detachment, thin and transparent retina, and formation of subretinal cords. In 40 eyes undergone scleral encircling and buclking, 36 (90%) had one-off successful operation, and the visual acuities over 0.05 were found in 77.5% of the whole eyes.ConclusionsScleral encircling and buckling procedures can be used to treat long-standing retinal detachment with subretinal proliferation with fairish cured rate. (Chin J Ocul Fundus Dis, 2005,21:150-152)
Objective To investigate the activation and role of signal transduction pathway of epidermal growth factor (EGF)-epidermal growth factor receptor (EGFR)-mitogen activated protein kinase (MAPK) in proliferation of human retinal pigment epithelial (RPE) cells. Methods Human RPE cells were stimulated with 0.1%，10% foetal calfserum (FCS) and EGF(0.1, 1, 10, 50 and 100 ng/ml)in 0.1% FCS Dulbeco′s modified Eagle′s medium (DMEM) and in 10% FCS DMEM for 3 days, respectively. Immunohistochemical staining and in situ hybridization were used to observe the expressions of EGFR protein and EGFR mRNA，respectively. Activation of MAPK was detected by immunohistochemical method with specific anti-phosphorylated ERK 1/2 antibody. Results The optimal concentrations of EGF were 10 ng/ml in 0.1% FCS DMEM and 1 ng/ml in 10% FCS DMEM. After 3 days of stimulation with EGF, phosphorylated ERK 1/2 staining was detectable in nucleus of RPE cells, whereas cells presented immunostaining for phosphorylated ERK 1/2 in the cytoplasm before stimulation. Conclusions EGF may improve the expression of EGFR protein and EGFR mRNA of RPE cells, and induced MAPK nuclear translocation in a concentration-dependent manner. EGF-EGFR-MAPK signal transduction pathway may play a key role in RPE cells proliferation, and serum exerts an important acceclerating function in the process. (Chin J Ocul Fundus Dis，2004，20：67-132)
Objective To clarify the relationship between inhibition of proliferation and cxpression of Ki-67 in cultured human retinal pigment epithelial(RPE) cells. Methods The cultured human RPE cells were treated with daunoblastina at a dose of 180 mu;g/L for 12h.Twenty-four hours later,DNA inhibiting rate was studied by using tritium-labelled thymidine deoxyribose(3H-TdR)incorporation assay.The expression of Ki-67 was evaluated by immunocytochemical staining technique and image analysis system.Flow cytometry was used to analyse cell cycle. Results DNA inhibiting rate was directly proportional to the dosage of daunoblastina.The proportion of the cells positive staining to Ki-67 in the control and the daunoblastina-treated group were 89.3% and 45.6%(Plt;0. 01),and the integral optical density values for expression of Ki-67 in the two groups were 68.1plusmn;6.2 and 27.3plusmn;5.5(Plt;0.01),respectively.The percen tage of cells in G2 phase of cell cycle increased from 8.9% to 29.5%. Conclusion G2 block was induced and poliferation was inhibited by daunoblastina in cultured human RPE cells.There is a relatively good correlation between Ki-67 immunostaining and inhibition of RPE cell proliferation. (Chin J Ocul Fundus Dis，2000，16：1-70)
Objective To investigate the effect of HGF on proliferation and migration in cultured human RPE cells. Methods Human RPE cells cultured in serum-free medium were treated with HGF(1，2，10，50，100 mu;g/L), and MT T assay was used to detect the growth of the cells; an in vitro wound healing model was used to count the number of cells that had entered the denudate area in RPE mig ration treated with HGF (1，2，10，50，100 mu;g/L) after 20 h. Results HGF(10,50,100 mu;g/L) increased proliferation rates of cultur ed human RPE (18.2 % to 34.8 %), and at a concentration of 50 mu;g/L on day 3 HGF induced the maximal increase of proliferation (Plt;0.01）; HGF showed effects on migration of 9.3 %, 113.0 %, 91.7 % and 50.3 % at the concentrations of 2,10,50, 100 mu;g/L, respectively. HGF stimulated t he best migratory response in RPE cells at a concentration of 10 mu;g/L (113 %). Conclusion HGF can induce the proliferation and obvious migration of RPE cells, consequently HGF was a mitogen and potent migratory factor for human cultured RPE cells. (Chin J Ocul Fundus Dis, 2001,17:307-310)
Purpose To investigate the expression of intercellular adhesion molecules ICAM-1 and Mac-1，in epiretinal membanes (ERM) of eyes wi th proliferative vitreoretinopathy (PVR)． Methods Twenty epiretinal membranes were obtained from eyes undergone vitrectomy for retinal detachment complicated with PVR and observed by immunohistochemical examination． Results Expressions of ICMA-1 and Mac-1 were observed in 18 and 15 membranes respectively．Expression of both adhesion molecules in 12 membranes． Conclusion The findings indicate that adhesion molecules might be involved in the development of PVR． (Chin J Ocul Fundus Dis，2000，16：71-138)
Objective To investigate the effects of glycation pro ducts on growth and cytosoic free calcium (［Ca2+］i) of bovine retinal capillary pericytes. Methods The changes of growth and ［Ca2+］i of bovine retinal pericytes,which were cultured in early glycation products of bovine serum albumin (EG-BSA) and advanced glycation end products of bovine serum albumin (AGE-BSA),were studied by counting cell numbers,MTT colorimetric assay,［3H］thymidine incorporating,and fluorescent indicator fura-2 acetoxymeth1 ester (Fura-2AM). Results The number of alive pericytes in groups of EG-BSA and AGE-BSA were 17.87plusmn;2.36 and 14.77plusmn;3.72 which comparing with their control groups (20.54plusmn;0.82 and 20.31plusmn;0.93)were de creased 13．00% and 27．00% (Plt;0.01) by counting cell numbers on a counting plate after four days.The results were 0.4619plusmn;0.0946 and 0.3884plusmn;0.1031 which comparing with their control groups (0.5236plusmn;0.0539 and 0.5227plusmn;0.0519)were decreased 12．00% and 25.70% (Plt;0.01) by MTT colorimetric assay.Amount of ［3H］thymidine incorporating in groups of EG-BSA and AGE-BSA were 39450.16plusmn;887 0.68 and 33667.85plusmn;10581.70 which comparing with their control groups (56373.63plusmn;2317.97 and 56542.04plusmn;1961.23)were decreased 30．00% and 40.40% (Plt;0.01).The ［Ca2+］i concentration of pericytes in groups of EG-BSA and AGE-BSA were (129.55plusmn;30.41) nmol/L and (179.71plusmn;56.69) nmo l/L which comparing with their control groups ［(79.70plusmn;6.94) nmol/L and (83.plusmn;6.39) nmo l/L］ were increased to 163．00% and 214．00%. Conclusion Both EG-BSA and AGE-BSA can inhibit the proliferation and DNA syntheses of retinal capillary pericytes,and increased ［Ca2+］i concentration in pericytes,especially the AGE-BSA. (Chin J Ocul Fundus Dis，2000，16：139-212)
Purpose To investigate the expression of the interleukin-6(IL-6)and tumor necrosis factor alpha(TNF-alpha;) in epiretinal membranes(ERM) of eyes with proliferative vitreoretinopathy(PVR). Methods Nineteen epiretinal membranes were obtained form eyes undergoing vitrectomy for retinal detachment complicated with PVR and observed by immunohistochemical methods. Results Expression of IL-6 and TNF-alpha; were observed in 12 and 15 membranes respectively with positive staining mostly in extracellular matrix of epiretinal membranes.Only one membrane showed positive to IL-6 intracellularly,and expression for IL-6 and TNF-alpha; simultaneously in membranes. Conclusion The findings indicate that IL-6、and TNF-alpha;might be involved in the development of PVR. (Chin J Ocul Fundus Dis,1998,14:219-221)
Objective To study the expression of receptor of advanced glycation end products (RAGE) in autogenous vein graft of streptozotocin induced diabetic rats and the inhibitory effects of aminoguanidine on intimal hyperplasia. Methods Sixty male Sprague-Dawley rats were randomly divided into three groups: aminoguanidine group, distilled water group and control group. Autogenous vein graft models were established in all groups. Streptozotocin was injected into abdominal cavity to induce diabetes in both aminoguanidine group and distilled water group, and they were intragastric administrated with aminoguanidine or distilled water, respectively before and after transplantation. Specimens were collected from autogenous vein graft 7 days and 14 days after surgery to undergo histological examination. At the same time, the level of serum advanced glycation end products (AGE) was tested. Western blotting and immunohistochemistry were used to detect the protein expression of RAGE and NF-κB p65. RAGE and NF-κB p65 mRNA were measured by reverse transcription-PCR. Results The mRNA and protein expressions of RAGE, NF-κB p65, the level of serum AGE and the intimal thickness of vein graft in distilled water group increased in comparison with those in control group 7 days and 14 days after surgery （P＜0.05). The level of serum AGE, mRNA and protein expressions of NF-κB p65 and the intimal thickness of vein graft in aminoguanidine group were lower than those in distilled water group （P＜0.05), and showed no significant difference compared with control group （P＞0.05). Conclusion The over-expression of RAGE in vein graft activats NF-κB in streptozotocin-induced diabetic rat, which has a close relation with intimal hyperplasia. Aminoguanidine can block the binding of AGE and RAGE by inhibiting the production of AGE, which will prevent intimal hyperplasia of vein graft.