Abstract:The introduction of highly active antiretroviral therapy (HAART) has greatly changed the pattern and natural history of ocular diseases of HIV-infected patients, resulting from the immune recovery and reduction of opportunistic infections. However, ophthalmic complication continues to be concern in AIDS even in the HAART era, especially in developing areas, where absolute majority of HIV-positive patients live. Lack of test facilities and experience, poor conditions of hygiene, different microbiological environment, absence of effective treatment etc., characterize the ophthalmic manifestation of HIV-infected patients in developing countries from that in developed regions and thus pose a great challenge to the ophthalmic treatment in developing area. Not only varied from region to region, ocular complications are distinctive between adults and children. At the same time, the side effects due to the application of HAART pose their own risks of ocular comp- lication and should, therefore, be given more resear

Abstract:AIM: To study the effects of 10g/L hydralazine eye drops on 35mg/kg NaIO₃ -induced degeneration in rat eyes. METHODS: Various doses of NaIO₃ and/or saline alone were injected into Brown Norway rats from hypoglossal vein. After 3, 7, 14 or 28 days of injection, ERG a-, b-, c-wave, fast oscillation (FO) and light peak (LP) were measured along with retinal colored pictures and fluorescein angiography(FA) taken. Some rats were chosen to study the histology of retinas by light microscopy and autofluorescence of retina flatmounts. Different concentrations of NaIO₃ were given to RPE-19 cells, and cell proliferation rate was measured. For hydralazine study, 35mg/kg NaIO₃ was injected into Brown Norway rat from hypoglossal vein. NaIO_{3 group was treated with saline alone after NaIO3 injection, 10g/L hydralazine+ NaIO3 group was treated with 10g/L hydralazine eyedrops after NaIO3 injection whereas normal group was treated with saline alone without NaIO3 injection. All eyedrops were instilled locally 3 times a day for 4 weeks and ERG c-wave was measured at the end of 2 and 4 weeks. RESULTS: After NaIO3 administration, the amplitude of all ERG waves fell markedly in large dose groups at 30, 40 or 60mg/kg NaIO3. Not many changes were observed in groups treated with <30mg/kg NaIO3. Some retinal necrosis appeared from 3 days post-injection (PI) in 30mg/kg NaIO3 group, which became more serious in larger dose groups or longer treatment time, but no apparent change was found in smaller dose groups. Similarly, on the retina flatmount, RPE monolayer showed necrosis from 3 days PI in the 30mg/kg NaIO3 and larger dose groups. On histological examination, no significant change was seen in 30mg/kg NaIO3 and lower concentration groups. In cell culture experiment, changes were found in RPE-19 cells proliferation rate with a concentration of NaIO3 at 30mg/L or higher. In hydralazine experiments, 4 weeks after injection of NaIO3, ERG c-wave fell markedly in NaIO3 group to 31% of control group(P <0.01). The ERG c-wave of hydralazine +NaIO3 group fell only to 50% of control group (P <0.05). This was a 61% reversal of the c-wave of NaIO3 treated group. CONCLUSION: Retinal pigment epithelium(RPE) degeneration induced by NaIO3 was both dose and time dependent. Around 30 to 40mg/kg NaIO3 would be the optimal to be used as a non-exudative age-related macular degeneration(AMD) rat model. Hydralazine may postpone the development of non-exudative AMD.}

Abstract:AIM: To evaluate the antioxidant activity of naringenin in human retinal pigment epithelium(ARPE-19) cells and human umbilical vein endothelial cells (HUVEC). METHODS: MTT assay was used to measure the viability and proliferation of ARPE-19 cells and HUVEC. RESULTS: Three and 10mg/L naringenin significantly increased the proliferation of ARPE-19 cells by 10.8% and 11.4%, respectively. Ten mg/L naringenin increased hypoxia-, 0.3mmol/L NaN₃^-, and 200umol/L H₂O₂ -induced damage of ARPE-19 cells by 55.2%, 69.2%, and 50.3%, respectively. One mg/L naringenin increased the viability of 50umol/L t-BHP-, and 30mg/L NaIO₃-treated ARPE-19 cells by 20.2% and 30.4% , respectively. Thirty mg/L naringenin also increased the proliferation of 50umol/L t-BHP-treated ARPE- 19 cells by 32.2%, and 1mg/L naringenin increased the proliferation of 30, 100 and 300mg/L NaIO₃-treated ARPE-19 cells by 30.3% , 10.3% and 18.5% , respectively. The reduction of HUVEC was 23.9%, 70.4% and 77.9% in the 3, 10 and 30mg/L naringenin-treated groups, respectively. Furthermore, 1 and 3mg/L naringenin increased hypoxiainduced damage in HUVEC by 10.7% and 13.1% , and 300mg/L NaIO₃-induced damage in HUVEC by 41.2% and 37.7%. Three mg/L naringenin increased 200 and 400umol/L H2O2-in-jured HUVEC by 20.1%and 21.5%, respectively. CONCLUSION: Naringenin increases the proliferation of ARPE-19 cells and inhibits the growth of HUVEC, and has potent antioxidant activity in ARPE-19 cells and HUVEC.

Abstract:AIM: To examine the expression of MMP-9 and TIMP-3 mRNA during choroidal neovascularization (CNV) in a murine model and to investigate the role of them in the development of CNV. METHODS: CNV was induced in C57BL/6J mice by intensive diode laser (810nm) photocoagulation (120mW, 75μm, 0.1s) of the fundus whereafter eyes were enucleated at 1, 3days, 1, 2, and 4 weeks. The MMP-9 and TIMP-3 mRNA expressions were analyzed using in situ hybridization and image analysis system. RESULTS: Both expression of MMP-9 and TIMP-3 mRNA had dynamic changes. For MMP-9, the expression was 1, 2, 4 wk>3d>1d (P <0.05), whereas TIMP-3 mRNA, 3d, 1, 2, 4 wk>1d (P <0.05). CONCLUSION: The imbalance between the changes of MMP-9 and TIMP-3 may accelerate the degrading of extracelluar matrix, and then be involved in the pathogenesis of CNV.

Abstract:AIM: To determine whether acellular porcine cornea stroma (APCS) could support the growth of the rabbit corneal cells in vitro . METHODS: APCS was prepared. The rabbit's corneal epithelium and stromal cells were cultured and seeded on APCS in vitro . The observation of phase contrast photograph and histological examination were performed. RESULTS: Histological examination showed the epithelium grew on the scaffold of APCS in 2-3 layers at 10th day. The stromal cells adhered to the surface of the scaffold after 24 hours and invaded into the interlaminar of the material at 5th day. CONCLUSION: These results indicate that APCS can support the growth and proliferation of the corneal epithelium and stromal cells in vitro .

Abstract:AIM: To explore the skills and characteristics of corneal neovascular model in rat induced by micropocket assay. METHODS: Nine eyes of nine Sprague-Dawley rats were studied. Pellets made of vascular endothelial growth factor (VEGF), poly-2-hydroxylethylmethacrylate and sucralfate were implanted into the corneal stroma no closer than 1mm from the limbus. Biomicroscopic features of corneal neovascular were observed on 1, 3, 5, 7th day after the implantation. RESULTS: On day 1 after operation, the limbal vessels were dilated, with no angiogenesis appeared On day 3, angiogenesis began to invade pericornea with a brush shape, the area of CNV was (2.23±0.59)mm². On day 5, new vessels reached the lower margin of pellet densely, and the area of CNV was (6.81±1.35)mm². On day 7, new vessels continued to elongate, parts of them extended as loops toward the pellet, and the area of CNV was (8.92 ±1.79)mm². Neither hyphema or other complications occurred. CONCLUSION: Corneal neovascularization induced by micropocket assay in rat grows steadily without complication, and is suitable for quantitative researches.

Abstract:AIM: To investigate the effect of β1-integrin overexpression on the apoptosis of rabbit corneal epithelial cells and the related mechanism. METHODS: The plasmid expressing β1-integrin-GFP fusion protein was constructed by polymerase chain reaction (PCR), and this plasmid (β1 group) or the empty vector (mock group) was transfected into rabbit corneal epithelial cells, respectively. The expression of β1-integrin-GFP fusion gene was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The adhesion of transfected cells to extracellular matrix (ECM) proteins was determined by adhesion assay. The apoptosis of rabbit corneal epithelial cells was assayed by Hoechst 33342 staining and DNA ladder. The phosphorylation of mitogen-activated protein (MAP) kinase was examined by Western blot. RESULTS: Rabbit corneal epithelial cells overexpressing β1-integrin-GFP fusion gene were successfully established. Compared with mock group, β1-integrin transfection significantly promoted the adhesion of rabbit corneal epithelial cells to ECM proteins such as laminin, fibronectin, collagen I and collagen IV. β1-integrin overexpression inhibited apoptosis and induced MAP kinase phosphorylation in rabbit corneal epithelial cells (P <0.05). CONCLUSION: These data suggest that overexpression of β1-integrin confers resistance to apoptosis in rabbit corneal epithelial cells, and MAP kinase pathway may play an important role in this process.

Abstract:AIM: To investigate the roles of feeder cells in stratification of murine corneal epithelial cells and build an ideal method to engineer stratified epithelial sheet. METHODS: Using contact feeder culture, separated feeder culture, compound feeder culture and culture without feeder cells by air-lifting method in Transwell chamber culture system, tissue engineered corneal epithelium was reconstructed. Corneal sheets were stained with hematoxylin and eosin (HE) for histological observation. The expression of p63 and keratin 19 (K19) and involucrin (IVL) was investigated by immunocyto- chemistry analysis. RESULTS: Stratification was limited to three to four layers in the contact feeder group, whereas separate feeder sheets were slightly more stratified. The compound feeder group produced a stratified epithelium with five to seven layers of cells. The group without 3T3 feeder cells formed only two to three layers of cells. Immunostaining images in the compound feeder group showed expression of progenitor markers p63 and K19 in the basal and suprabasal layer, as well as differentiation marker involucrin in all layers. CONCLUSION: The remarkable stratification as well as the limbal phenotype makes the compound feeder system a candidate tool for cultivating transplantable epithelial sheets.

Abstract:AIM: To observe the effects on human keratocytes by cationic liposome Lipofectamine^TM2000 (LF2000), to investigate the efficiency and safe range applied in human keratocytes, and establish basis for gene therapy of human keratocytes. METHODS: Human keratocytes cultured in vivo within 3 to 5 passages were used in experiment after being identified. The effects on proliferation of cultured human keratocytes by LF2000 with different concentrations and time were evaluated By MTT; the effects of LF2000 on the survival rate and its relation with 5,10,20,40,80mg/L concentration and time were detected by trypan blue staining. RESULTS: LF2000's effects on human keratocytes were related with concentration and time. The cellular proliferation and survival rate declined when concentration of LF2000 was above certain level, and this effect increased as time became longer. LF2000 had no effect with concentration under 40mg/L for 24 hours. CONCLUSION：LF2000 did not cause cytotoxicity during a concentration range“tested”, and it is hoped to play an important role in gene therapy of human keratocytes.

Abstract:AIM: To study the feasibility of recombinant adeno-associated virus (rAAV) as a vector to transfer the green fluorescent protein (GFP) gene as a target gene into rabbit retina. METHODS: Intravitreal injection of rAAV-gfp was performed in either eye for each rabbit with the other eye taken as control. At the 3rd, 7th, and 14th day after injection, the eyeballs were removed, and the retinas were flat-mounted on glass slides to inspect the retinal fluorescence, respectively. RESULTS: After intravitreal injection of rAAV-gfp, the presence of fluorescent spots in the cytoplasm of retinal cells indicated that GFP gene was efficiently transferred and expressed in the rabbit retina. CONCLUSION: Recombinant adeno-associated virus is a reliable and simple vector for transferring target gene, e.g., GFP gene, to the retina.

Abstract:AIM: To culture astrocytes from human donor eyes in order to understand the function of astrocytes in remodelling events in the glaucomatous optic nerve head (ONH). METHODS: Primary cultures were prepared by explantation of human ONH tissue in order to get astrocytes. Laminar criborsa (LC) cells were prepared concurrently for com- parison. Astrocyte cultures could be separated from LC cells by selecting medium. Similar procedures were used for LC. RESULTS: Primary cells grew from human optic nerve head explants 4-8 weeks after explantation. Astrocytes had different morphologies and growth characteristics from LC cells. Type 1B astrocyte cells could grow in medium without FBS. Purified cultures were obtained by second passage and could be harvested by third to fifth passage, which were prepared to use for further study, including being characterized by positive glial fibrillary acidic protein (GFAP) and neural cell adhesion molecule (NCAM) staining. CONCLUSION: Precise dissection of fragment is the most important step to get clear explants for primary culture. Economic and rapid method could be useful to select cells by different mediums, which will help us to get more purified cells for further study.

Abstract:AIM: To evaluate the extent of significant posterior capsular opacification (PCO) after implanting polyme-thylmethacrylate (PMMA) and soft acrylic intraocular lenses. METHODS: A total of one hundred patients (110 eyes) undergoingintraocular lens(IOL) implant surgerywererandomly divided into two groups based on the type of IOL implanted: Group I: AcrySof (SA60AT) and Group II: PMMA (LX10BD). The density of posterior capsular opacification (PCO) was assessed 3, 6, 12 and 18 months after surgery by taking best corrected visual acuity (BCVA) and the digital photographs. RESULTS: At the end of 3 months postoperatively, the PCO density in the Group I (PMMA) increased significantly (3.6%) while no increase was found in the Group II (Acrylic). By the end of 18 months follow-up, the incidence of significant PCO was found to be less in the acrylic group (14.5%) as compared to the PMMA group (34.5%).On the basis of density, the PCO was more extensive with the PMMA lens than the AcrySof lens, which led to more severe visual loss. CONCLUSION: The intraocular implantation of the AcrySof IOL helps to reduce the incidence of PCO to a greater degree as compared to the PMMA IOL.

Abstract:AIM: To identify the predisposing factors, causative fungi and to improve the facilities for the laboratory diagnosis of fungal keratitis. METHODS: Two hundred and forty eyes of 240 patients of clinically suspected microbial corneal ulcer were included in the study. Data was collected through history and slit lamp examination. Using standard techniques, corneal scraping was performed.A portion of each scraping was examined by direct microscopy for the presence of fungi, bacteria and acanthamoeba by using 100g/L potassium hydroxide and also by Gram and Giemsa stainings. Another portion was inoculated directly on the surface of solid media such as blood agar, Mac-Conkey agar, chocolate agar and Sabouraud agar in C-shaped streaks for culture. RESULTS: A total of 240 patients with suppurative corneal ulcer were enrolled in the study, out of which fungi were identified in 84 (35%) patients. Of these, 48 (57.1%) were males and 36 (42.9%) females. The age ranged between 22-80 years. Sixty patients (71.4%) belonged to rural population and twenty four (28.6%) to urban population. Trauma with vegetative material was the most frequent cause noted in eighteen (21.4%) patients. Peak incidence was in the months of October-November. Out of 84 eyes with fungal keratitis, fungi alone were the etiologic agents in 74 (80.10%) cases and bacteria with fungi were identified in 10 (11.90%). The most frequently isolated organism was Candida albicans which was found in 66 (78.6%) patients. CONCLUSION: Fungal keratitis is the leading cause of infective corneal ulcer and Candida albicans being the most commonly isolated pathogen in the patients belonging to Southern Pakistan. The direct microscopic examination with potassium hydroxide 100g/L method is a simple, rapid, inexpensive and reliable method in the diagnosis of this infection.

Abstract:AIM: To determine the rate of bacterial colonization in hydrogel disposable contact lenses and solutions and to identify the contaminating organisms. METHODS: A cross sectional study with purposive sampling was carried out. One hundred hydrogel contact lenses were collected from wearers among undergraduate students of Health Campus, University Sains Malaysia. All lenses and storage solutions were sent for microbiological culture and gram staining. RESULTS: The majority of study participants were females (98%). The mean age was 21.36±1.63 years. Forty-one subject participants (82%) showed positive bacterial colonization of the lenses. From storage solutions 32% yielded positive colonization by bacteria. The most common organisms were coagulase negative staphylococcus, Staph aureus and streptococci while Pseudomonas sp. and Serratia sp. were isolated more frequently from contact lenses. CONCLUSION: Contact lens wearing is potentially dangerous as a result of high rate of bacterial colonization of the lenses and its storage solutions. Extreme precaution and adherence to strict hygienic practice is recommended during lens handling and wearing.

Abstract:AIM: To present the outcome of anterior single flap external dacryocystorhinostomy (DCR) in Sudanese patients. METHODS: The data of 200 consecutive patients were retrospectively analyzed, who had anterior single flap external DCR with a minimum of one year follow-up. The surgeries were performed by the same surgeon (the first author) and patients were followed up for one year postoperatively. Presence of epiphora at the end of one year and no response to syringing and probing was considered failure. RESULTS: The mean age of the study sample was 29.7 years (ranged 4-65 years). The ratio of male to female was 1∶2. The success rate was 98%. Failure was reported in 4 patients, two of them were traumatic cases with preexisting orbital disfigurement. CONCLUSION: This study adds on to the evidence of the usefulness of anterior single flap DCR. Although it is simpler and easier to master the technique, this procedure showed a success rate comparable to that of the conventional method in literature.

Abstract:AIM: To investigate the conditions of traumatic visual loss of inpatients in Yazd, Iran from 2005 to 2006, and to explore the possible causes and preventive methods. METHODS: We retrospectively analyzed data on all patients with eye injuries (70 cases) hospitalized at Shahid Sadoughi Hospital between August 2005 and August 2006, including age, gender, causes, type of injury, time of initial treatment, visual acuity during admission and discharge, surgical procedures, and final outcome. Furthermore, standardized international classification of ocular trauma (Birmingham Eye Trauma Terminology) was used for eye injury categorization. · RESULTS: A total of 70 inpatients with ocular injuries were selected by simple approach. The male/female ratio was 2.8 to 1. The most frequent causes of eye injury were metallic objects (34.3%) especially in age group of ≤10 and ≥30 years old, accidents (22.8%) and assaults (17.2%). Waiting time to initial treatment was 6-24 hours in most cases (51.4%). Trauma was blunt in 38.6% and penetrating in 61.4%. The most and the least final visual acuity in blunt and penetrating trauma was LP (51.8%, 41.8%) and HM (7.5%, 4.7%) respectively. Surgery outcome was globe saving with acceptable visual acuity in 72.9% and enucleation in 24.2%. CONCLUSION: With regard to the high incidence of ocular trauma and consequent severe visual loss, parents' attention, eye safety protection and early treatment should be considered as final preserving globe in most accidents.

Abstract:AIM: To evaluate the prevalence of glaucoma, treatment patterns and patient attitudes in Ethiopia. METHODS: A survey was administered to glaucoma patients in hospitals in Addis Ababa, Ethiopia. RESULTS: Of the 415 qualified patients, exfoliative glaucoma (17%) was most commonly found in the Gurage population(33%), whereas chronic angle-closure glaucoma (5%) was found in the Amhara/Tigre (6%) population (P =0.006). Patients were treated with an average of 1.4±0.8 medications: 52% admitted recent noncompliance and 36% had undergone filtering surgery. Patients believed their physician was trying to help them (93%). God wanted them to receive treatment (89%) and their spouse was sympathetic to their disease (82%). Patients noted their community was unaware of their condition (87%) with Muslims [most common in the Gurage population (31%, P <0.0001)] reporting this the most(25%,P =0.01).Amhara/Tigre patients strongly believed their doctors were concerned about them (94%, P = 0.04). CONCLUSION: The prevalence of glaucoma type varies among ethnic groups in Ethiopia with exfoliation more common in the Gurage population and chronic angle-closure glaucoma more frequent in the Amhara/Tigre population.

Abstract:Surgical treatment of glaucoma has been classified as cyclodestructive (reducing inflow) or filtering (increasing outflow). Cyclodestructive procedures have traditionally been reserved for eyes with poor visual prognoses and refractory glaucomas including post-trauma, aphakia, congenital and developmental glaucoma. Since Uram described the first use of endoscopic cyclophotocoagulation (ECP) in 1992, short and long-term outcomes for ECP have been promising. In the present article, we conduct a Pubmed search and review of published English literature on endoscopic cyclophotocoagulation and comparison with limited results in a single Singapore ophthalmic tertiary hospital. Safety and efficacy of ECP and combined phacoemulsification-ECP procedures in treatment of pediatric and adult glaucomas of various etiologies and severities is reported. Local short-term unpublished results from a single Singapore tertiary ophthalmic service is reported and concurs with previously published results.Published reports and current experience with ECP has demonstrated that ECP with direct visualization of the target tissues avoids the complications associated with blind trans-scleral cyclophotocoagulation by applying optimum energy to target tissue ciliary epithelium with endoscopic visualization and infrared laser wavelength application. Significant financial barriers exist to introducing this service. It is safe and effective in controlling IOP and reducing reliance on anti- glaucoma medications. Widespread acceptance and use of this technique awaits large-scale randomized controlled studies.

Abstract:Idiopathic intracranial hypertension (IIH) is the syndrome of raised intracranial pressure (ICP) without clinical, laboratory or radiological evidence of intracranial pathology. IIH is a relatively rare disease but rapidly increasing incidence is reported due to a global increasing incidence of obesity. Disease course is generally said to be self-limiting within a few months. However, some patients experience a disabling condition of chronic severe headache and visual disturbances for years that limit their capacity to work. Permanent visual defects are serious and not infrequent complications. The pathophysiology of IIH is not fully understood yet. Advances in neuroimaging techniques have facilitated the exclusion of associated conditions that may mimic IIH. No causal treatment is yet known for IIH, and existing treatment is symptomatic and rarely sufficient. The aim of this review is to provide an updated overview of this potentially disabling disease. Theories of pathogenesis, diagnostic criteria and treatment strategies are discussed.

Abstract:AIM: To report a rare case of mesenchymal chondrosarcoma in the orbit and to explore its clinic manifestations, pathologic characters, management and prognosis. METHODS: We report a case of mesenchymal chondrosarcoma of the orbit. The clinical materials, including ophthalmological examination, computed tomography scan of the orbit, histopathology and immunohistochemistry of the biopsy specimen were reported, and its pertinent literatures were reviewed. RESULTS: A 36-year-old female was seen with proptosis and decreased vision. Histopathology demonstrated an admixture of undifferentiated mesenchymal cells and islands of mature hyaline cartilage. Immunohistochemical studies revealed positivity for vimentin and S-100, which was consistent with the diagnosis of mesenchymal chondrosarcoma. · CONCLUSION: Mesenchymal chondrosarcoma in the orbit is extremely rare malignant tumor. Multi-modality treatments (surgery, chemotherapy and radiotherapy) may lead to long-term survival.