Objective To establish a method for primary culture of iris pigment epithelial cells (IPE). Methods Enzyme Assisted microdissection was used to isolate and cultivate the IPE cells. An identification was made with microscopic and immunohistochemical observations. Results IPE were successfully cultured and showed on differences with RPE in primary culture and subculture. Conclusion Enzyme Assisted microdissection is a reliable and quick method for the isolation of IPE.

Objective To investigate the effects of genistein with different concentration on proliferation and apoptosis of cultured human retinal pigment epithelial (RPE) cells in vitro. Methods The effect of genistein with the concentration of 5,10,25,50,75,and 100 mg?L -1 on the proliferation of cultured RPE was examined by tetrazolium salt (MTT) assay and AgNORs staining. The cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick-end labeling (TUNEL) methods, in the mean time, the morphologic changes of cell apoptosis were observed by light microscopy and transmission electronic microscopy, the results of which were compared with the normal RPE cells. Results Genistein with the concentration of 25, 50, 75, and 100 mg?L -1 had a dose-dependent and time-dependent antiproliferative effects on RPE cells with the inhibitory rate of 12.0%-64.6% (P

To detect the effects of adrenaline and dexamethasone on proliferation of the lens epithelial cells and discuss the mechanism, the central area 5～7mm in diameter of the anterior capsule was obtained in cataract surgery (phacoemulsification, Phaco), and the specimens were divided into three parts, and were cultured in vitro and treated with 10 -6 mol/L adrenaline and dexamethasone for 48 hours. And then the positive area ratio of proliferating cell nuclear antigen (PCNA) in immunohistochemistry was estimated by medical color image analysis. The data were analyzed with the statistical software. The results indicated that the 10 -6 mol/L adrenaline and dexamethasone had obvious inhibition on LEC proliferation. The positive area ratio of PCNA was (2 614?0 922)%( t =5 594, P =0 0000) and (3 338?0 838)%( t =3 361, P =0 0013). It is suggested that adrenaline and dexamethasone could inhibit the proliferation of LEC and could be used to treat PCO after cataract surgery. The experiments provide the scientific basis for selection of drugs to prevent PCO after cataract.

Objective To observe the influence of the expression of CD18 on the neutrophile and the leukocyte adhesion to retinal vascular endothelium by hypoxia-inducible factor-1 alpha (HIF-1α) in early diabetic retinopathy rats. Methods Male Sprague-Dawley rats received intraperitoneal injection of streptozotocin to induce diabetes model. 18 diabetic rats were divided into 3 groups randomly after 2 months of diabetes induction, including diabetic group (group B), HIF-1α anti-sense oligonucleotides (ASODN) injection group (group C) and HIF-1α sense oligonueleotides (SODN) injection group (group D), the age and weigh matched health rats were chosen as control group (group A), with 6 rats in each group. Then group A and B rats received 5 % glucose solution caudalis veins injection, group C and group D rats received HIF-1α ASODN and HIF-1α SODN caudalis veins injection, respectively(0.25 mg/kg). The level of CD18 on the neutrophil isolated from the peripheral blood was measured by flow cytometry. Retinal leukostasis was quantified with acridine orange leukocyte fluorography. Results The percentage of CD 18 positive neutrophil cell was (44.93±3. 60)% in group B, (18.66±1.52)% in group A, (31.66±4.72)% in group C,(51.00±5.66)% in group D. Compared with each other groups,the differences are statistically significant (F= 42. 46, P<0.001). The number of positive staining cells of retinal leukocyte was (46.16±10.68)in group A, (133.83±20.43)in group B, (99.83±9.28)in group C, (121.33±10. 23) in group C. Compared group B with group C,the number of positive staining cells raised about 2.89 times;compared group B with group C and D,the differences are statistically significant (P= 0.12, 95% confidence interval -3.69～28. 69 ). Conclusions In vivo, HIF-1α can decreased the expression of CD18 on neutrophils from diabetic rats' peripheral blood and the collection of retinal leukos- tasis in the diabetic animals. HIF-1α may serve as a therapeutic target for the treatment and/or prevention of early diabetic retinopathy.

Objective To investigate the effects of cytokines on the expression of syndecan-1 in cultured human retinal pigment epithelial (RPE) cells and the signal transduction pathway. Methods Reverse transcription polymerase chain reaction and immunofluorescence staining were used to detect the expression of syndecan-1 mRNA and protein in normal RPE cells. The expression of syndecan-1 in RPE cells stimulated by different cytokines was detected and quantitatively analyzed by image process of immunofluorescence. The stimulation included 7 and 35 ng/ml tumor necrosis factor (TNF)-? for 24 hours, 1 and 6 ?g/ml lipopolysaccharide (LPS) for 11 hours, 7 ng/ml TNF-? for 0 to 24 hours (once per 2 hours, and 13 times in total), and 30% supernatant of monocyte/macrophage strain (THP-1 cells) for 3, 14 and 43 hours. The effect of 30% supernatant of THP-1 cells was assayed after pretreated by PD098059 [the specific inhibitor of extracellular signal regulated kinase(ERK) 1/2] for 2 hours. After exposed to 30% supernatant of THP-1 cells for 3 hours and treated by 0.25% trypsin for 5 minutes, RPE cells attaching was evaluated by methyl thiazolyl tetrazolium assay. Results In normal human RPE cells, expressions of syndecan-1 mRNA and protein were detected, and strong syndecan-1 positive yellowish green fluorescence was found in the cell membrane and cytoplasm while light green fluorescence was in the nucleus. As the concentration and stimulated time of TNF-? or LPS increased, the fluorescence intensity decreased(P

0.05).Statistical analysis showed that there was a correlation between the expression of CTGF and TGF-?RⅡ,FN,and collagen Ⅰ and Ⅲ protein,respectively. Conclusions The expression of CTGF and TGF-?RⅡ protein is up-regulated in PRM of PVR,which suggests that the activation of TGF-?RⅡ is involved in the production of CTGF,and CTGF is closely related to the production of ECM and play an important role in the pathogenesis of PVR.

Objective To investigate the expression of hepatocyte growth factor receptor (HGFR) in epiretinal membranes (ERM) of eyes with proliferative vitreoretinopathy (PVR) and cultured retinal pigent epithelium (RPE) cells. Methods Fifteen human epiretinal membranes were obtained from eyes undergone vitrectomy for rhegmatogenous retinal detachment complicated with PVR and observed by immunohistochemical examination to study the expression of HGFR. Using the immunohistochemical technique to evaluate the expression of HGFR in cultured RPE cells. Results In 6 membranes of PVR grade C, HGFR were expressed in 5/6, and 7 cases were detected in 9 membranes of PVR grade D.RPE cells express readily detectable levels of HGFR. Conclusion The findings indicate that HGF might be involved in the formation of epiretinal membranes in PVR.

Objective To establish a hemorrhagic retinal detachment (HRD) model for the study of the damage and treatment of HRD. Methods Fourteen rabbits (28 eyes) were divided into the HRD (12 eyes) and control (16 eyes) group randomly. Autologous anticoagulated blood (0.2 ml) was transvitreally injected into the rabbits′ subretinal space with a special glass micropipette in HRD group (12 eyes); while 0.2 ml saline with or without heparin sodium (2.5 U/ml) was respectively injected into subretinal space respectively of the rabbits in heparin saline control group (6 eyes) and saline control group (3 eyes); furthermore, another 2 control groups, i.e.,pseudo injection group (3 eyes, single retinal puncturing without subretinal injection) and normal group (4 eyes of 2 normal rabbits) were also set. The conditions of the occurrence and representation of the retinal detachment (RD) were observed and analysed by means of ophthalmoscopy, optical coherence tomography (OCT) and ultrasound A and (or) B scan examinations in the subsequent 28 days after the operation. Results After the operation, HRD occurred in all eyes of the rabbits in HRD group. The area of HRD extended from 10 to 12 disc diameter(DD). The obvious elevation of RD maintained to 14 days, and the residual subretinal hemorrhage was still observed till 28 days. The obvious RD of the rabbits in heparin saline and saline control group was only kept for no more than 12 hours. The retinal puncture hole in pseudo injection group disappeared 2 days after the operation, and there was no change in retina of rabbits in normal control group. Conclusion It is convenient, practical and effective to establish a HRD model by means of transvitreal subretinal injection of autologous anticoagulated blood.

Purpose To investigate bax expression and induction of apoptosis in normal cultured retinal pigment epithelium (RPE) cells . [WT5”HZ]Methods [WT5”BZ]Cultured human RPE cells were transfected by P MDNA3 hbax ,which incoded the whole bax gene and may be induced by Zn 2+ under the MTII promoter, through lepofectin mediated protocol.The tested RPE cells were divided into three groups of A, P MDNA3 hbax transfected ;B,P MDNA3 (nude vector) transfected and C,normal RPE cells.After transfection, DNA gel electrophoreses were performed ,the tested RPE cell cycles were analyzed with flow cytometry (FCM). Results The gel electrophoretogram showed DNA ladder phenomenon,FCM confirmed the apoptosis of RPE cells P MDNA3 bax transfected , consisting of significant apoptotic peak sited before the G 1 phase and the apoptotic rate was 36%. Conclusion The foreign bax gene can be effectively conducted into the RPE cell through lepofectin mediated protocol and induced expression . The foreign bax overexpression may induce the cultured human RPE cell susceptibility to apoptosis.

Objective To evaluate the visual results,surgical technique and safety of secondary intraocular lens (IOL) implantation in aphakic eyes following vitrectomy and lensectomy for complicated ocular trauma or retinal detachment. [WT5”HZ]Methods The clinical records of 32 cases (32 eyes),received these surgeries during November 1996 and December 1999,were reviewed retrospectively.During the secondary operation,intraocular infusion through the pars plana was performend and the type of IOL was chosen based on the integrity of lens capsule. [WT5”HZ]Results The study included 30 eyes suffering from trauma (foreign bodies in 15 eyes,penetrating injury with traumatic endophthalmitis and with vitreous hemorrhage in 6 eyes respectively,blunt trauma with lens dislocation in 3 eyes),and 2 eyes with primary retinal detachment.Those eyes all received vitrectomy,lensectomy,and/or removal of foreign bodies and corneal suture.The interval of two operations ranged from 1 to 16 months with an average of 6.8?3.7 months.Posterior chamber IOL was implanted in the ciliary sulcus in 25 eyes with a whole or 2/3 of lens capsule,trans scleral suture fixation of IOL in 5 eyes,anterior chamber IOL and IOL with artificial iris in one eye respectively.Silicone oil was removed in 5 eyes during the secondary operation.Post operative visual improvement was achieved in 29 eyes.Main complications were corneal edema and low intraocular pressure after operation. [WT5”HZ]Conclusion Intraocular infusion and proper IOL implantation during the secondary operation following vitrectomy can provide selected aphakic eyes with better visual recovery.