Volume 43 ·  Issue 05 ·  2007

Since the 21st century, the ophthalmology research in China has made great progress in all aspects, and many fields have reached the international advanced level. However, there is still a gap between the ophthalmology research in China and the western developed countries, and one of the reasons is the lack of original innovation and interdisciplinary in-depth research. In order to further improve the ophthalmology research level in China and narrow the gap with the developed countries, the workers engaged in ophthalmology research need to accelerate the introduction, digestion, absorption and integration of research results and new technologies from other disciplines, so as to produce more original and innovative research. In the "post-genome" era, it is difficult for a single discipline to solve such complex life science problems, and the interdisciplinary and integration of multiple disciplines bring opportunities for ophthalmology research, especially the newly emerging disciplines such as bioinformatics and systems biology, which will help to promote the change of thinking mode and research means of ophthalmology scientific research.

The 3rd Global Chinese Ophthalmology Congress and the 11th National Ophthalmology Congress of Chinese Medical Association were held in Beijing International Convention Center from August 31 to September 4, 2006. Chinese ophthalmologists and visual researchers from all over the world exchanged extensively the achievements of basic and clinical research in ophthalmology. At the closing ceremony of this academic conference, the Visual Physiology Group reported the achievements and progress made by the Visual Physiology Group to the conference under the title of "Pioneering and Innovating in Interdisciplinary Exploration and Research".

Objective to construct a lentiviral vector that enhances the co-expression of green fluorescent protein (EGFP) and the suicide gene herpes simplex virus thymidine kinase (HSV-tk) regulated by the broad-spectrum promoter (CMV) of human cytomegalovirus, and to obtain an efficient lentiviral transgenic platform. Methods HSV-tk gene was inserted into the lentiviral vector Lenti-ires-EGFP by restriction endonuclease digestion and T4 DNA ligase ligation, and the lentiviral vector Lenti-CMV-HSV-tk-EGFP was constructed. The successfully constructed lentiviral tri-plasmid system was transfected into packaging cells-293T derived from human embryonic kidney cell line by calcium phosphate precipitation method. The virus particles were collected 28 h later, and the virus titers were measured. Human lens epithelial cell lines, retinoblastoma cell lines, neuroblasts cell lines and Hela cells were infected, and the expression of EGFP was observed under fluorescence microscope. The expression of HSV-tk and EGFP was detected by RT-PCR. Results the constructed lentiviral vector Lenti-CMV-HSV-tk-EGFP could transfect various cells efficiently and stably expressed. The results of RT-PCR showed that Lenti-CMV-HSV-tk-EGFP infected cells could co-express HSV-tk and EGFP. The transfection efficiency of Lenti-CMV-HSV-tk-EGFP was close to 100% when the degree of complex infection (MOI) was 100, and the transfection efficiency could still be maintained after 6 months of subculture. Conclusion TK suicide gene was successfully constructed to transfect many kinds of cells

Objective to investigate the role of glycosylation end product (AGE) in diabetic retinopathy (DR). Methods Cultured bovine retinal capillary pericytes were co-cultured with different concentrations (0.47, 1.88, 7.50 μ mol/L) for 4 days. The effects of apoptosis, caspase-3 activity and caspase-3 inhibitor Z-DEVD-fmk on pericyte apoptosis and the ratio of apoptosis regulatory gene Bcl-2/Bax were detected respectively. Results AGE could induce apoptosis and increase the activity of caspase-3 in cultured bovine retinal capillary pericytes in a dose-dependent manner (r =0.867, P<0.01), while selective caspase-3 inhibitor Z-DEVD-fmk could significantly inhibit apoptosis and increase the ratio of apoptosis regulatory gene Bcl-2/Bax under AGE. Conclusion Apoptosis is one of the mechanisms of selective loss of retinal capillary pericytes in DR, and the increase of caspase-3 activity is the cause of apoptosis induced by AGE

Objective to investigate the expression changes of Occludin and glial fibrillar acidic protein (GFAP) in the retinal endothelial cells of diabetic rats and their relationship with blood-retinal barrier (BRB). Methods the rat diabetic model was established by intraperitoneal injection of streptozotocin. After 1, 3 and 6 months, Evans blue (EB) was injected to evaluate the morphological changes of blood-retinal barrier destruction. The expression changes of Occludin and GFAP were observed by immunofluorescence histochemistry. Results At 1 month, the expression of GFAP in the nerve fiber layer and ganglion cell layer of the rat retina was significantly increased, and the linear fluorescence arrangement of Occludin was disordered, but the fluorescence intensity was not weakened or interrupted. The number of GFAP-positive Müller cells gradually increased at 3~6 months, and the expression of Occludin was weakened and the range of continuous interruption was continuously expanded. EB injection showed a synchronous development trend of blood-retinal barrier damage. Conclusion The activation of astrocytes in the early stage of diabetic retinopathy may play an important role in maintaining BRB function.

Objective to establish the expression profile of fibrin in normal lens of adult male.Methods the total proteins of lens fibers were preliminarily separated by polyacrylamide gel electrophoresis, and the peptides digested in gel were identified by reversed phase liquid chromatography tandem mass spectrometry.According to the matching rate of high abundance ions, the continuity of matched ions and the random score, the results were screened by SEQUEST and AMASS softwares.The same sample was repeated for three times, and the proteins with confidence>98%, two or more qualified spectra and not less than two experiments were identified as positive results. Results A total of 60 proteins were identified in the laboratory, and 22 lens proteins were identified in previous lens proteomic studies.Conclusion This study established the expression profile of fibrin in normal lens of adult male.The combination of polyacrylamide gel electrophoresis and liquid chromatography tandem mass spectrometry is a simple and highly sensitive proteomic method, It is suitable for studying the changes of lens fiber protein with age and cataract formation.

Objective to study the recovery of binocular vision in early patients after multifocal intraocular lens (MIOL) implantation. Methods 31 cases (46 eyes) of cataract patients were implanted with MIOL as MIOL group, including 16 cases undergoing monocular surgery in Ms group and 15 cases undergoing binocular surgery in Md group. 32 cases (47 eyes) of cataract patients were implanted with monofocal intraocular lens (SIOL) as SIOL group, including 17 cases undergoing monocular surgery in Ss group and 15 cases undergoing binocular surgery in Sd group. Three weeks after surgery, the binocular vision of patients were observed, namely, homoscopic function, fusion function, distant and near stereoscopic function, visual acuity and visual symptoms, etc. Results All patients had good homoscopic function and fusion function, and the difference between the two groups was not statistically significant (P>0.05). The patients with distant stereoscopic function accounted for 38.7% (12/31) in MIOL group and 40.6% (13/32) in SIOL group, and the difference between the two groups was not statistically significant (P>0.05). The patients with close stereoscopic function accounted for 41.9% (13/31) in MIOL group and 15.6% (5/32) in SIOL group There was no significant difference in stereoscopic function between Ms and Md, Ss and Sd (P>0.05). Only 2 (6.5%) patients with MIOL implantation in one eye complained of glare at night, while the other patients had no obvious discomfort, and the degree of satisfaction was higher. 80.6% (25/31) of MIOL group and 56.2% (18/32) of SIOL group complained of significant improvement in stereoscopic vision such as picking dishes at meal, flushing hot kettles, going up and down stairs, and the difference between the two groups was statistically significant (P>0.05). Conclusion Stereoscopic function of patients in the early stage after MIOL implantation has reached the normal level of the elderly; Moreover, the recovery of stereoscopic function of monocular and binocular implants is comparable. The recovery of near-range stereoscopic function in MIOL group is faster than that in SIOL group. MIOL has obvious advantages in providing patients with good whole-course vision and stereoscopic function.

Objective to investigate the characteristics of ocular manifestations in patients with human immunodeficiency virus (HIV) infection, so as to provide evidence for early diagnosis and timely treatment. Methods The clinical data of 278 patients with HIV infection in Central African Republic from September 2002 to July 2004 were retrospectively analyzed, and the course, symptoms, signs, systemic conditions and the causes of the first diagnosis were analyzed.Results 102 cases (36.7%) of HIV infection had ocular lesions, accompanied by chronic systemic wasting symptoms and signs, and CD4 cell value decreased.20 cases (7.2%) of them took ocular lesions as the causes of the first diagnosis. Ocular lesions were opportunistic infection and sarcoma formation, followed by HIV retinopathy, uveitis, extraocular muscle palsy, ocular herpes zoster and eyelid Kaposi's sarcoma, etc. HIV retinopathy is not specific, and it is manifested as retinal hemorrhage, cotton plaque, vascular occlusion, retinal necrosis, retinal detachment and optic nerve atrophy, etc. Conclusion A variety of ocular lesions may occur after HIV infection, Severe retinal and uveal lesions are important factors that cause blindness in patients. The number of HIV infected patients in China is increasing day by day, and their early symptoms and signs should be paid close attention by ophthalmologists.

Objective to study the correlation between primary open-angle glaucoma (POAG) and primary closed-angle glaucoma (PACG) patients and translipoprotein E (APOE) genes in Northeast China. Methods Polymerase chain reaction amplification (PCR) and restriction fragment length polymorphism (RFLP) were used to type APOE genes in 36 patients with POAG, 69 patients with PACG and 57 controls, and the frequencies of APOE alleles and genotypes in each group were calculated respectively. Results the frequencies of ε 3/ε 4 genotypes in POAG and ε 2/ε 4 genotypes in PACG (41.7% and 43.5%, respectively) were significantly higher than those in the control group (14.0% and 21.1%, respectively, P<0.05); The epsilon 4 allele frequency of APOE in POAG and PACG groups (37.5% and 39.2%, respectively) was significantly higher than that in the control group (17.5%, P<0.05), and the epsilon 2 allele frequency in POAG group (8.3%) was significantly lower than that in the control group (15.8%, P<0.05). Conclusion the epsilon 4 allele of APOE may be a risk factor of primary glaucoma, and epsilon 2 allele may have a certain protective effect on the occurrence of POAG.

Objective to observe the efficacy and safety of modified viscoelastic Schlemm tube incision (VCO) and trabeculectomy (TE) in the treatment of primary open-angle glaucoma.Methods A prospective randomized controlled method was used to treat 100 patients with primary open-angle glaucoma who had inadequate intraocular pressure control with drugs. The patients were randomly divided into 50 cases (50 eyes) in modified VCO group and 50 cases (50 eyes) in TE group. The average follow-up period was 28 months.Results One month after operation, the average intraocular pressure of patients in modified VCO group and TE group was (11.22+4.34) and (12.35+3.79) mmHg (1 mmHg =0.133 kPa), respectively. The success rate of operation (intraocular pressure<21 mmHg without any intraocular pressure lowering drugs after operation) was 98.0%; At 12 months after operation, the mean intraocular pressure of the two groups was (14.50±3.22) and (16.58±4.73) mmHg, respectively, and the success rate of operation was 87.5% and 70.0%, respectively, and the difference between the two groups was statistically significant (P<0.05); At 24 months after operation, the mean intraocular pressure of the two groups was 16.0 and 15.4 mmHg, respectively, and the success rate of operation was 76.1% and 67.3%, respectively, and the difference between the two groups was not statistically significant (P>0.05); The early complications such as shallow anterior chamber, choroidal detachment and cystoid filtering vesicles were significantly more in TE group than in modified VCO group, and the difference between the two groups was statistically significant (P<0.01). Conclusion In the treatment of primary open angle glaucoma, the success rate of modified VCO group was higher than that of TE group, and the early postoperative complications were relatively few.

Objective to observe the morphological characteristics of rabbit limbal epithelial cells cultured on human amniotic membrane in vitro and the formation of junction complexes with amniotic basement membrane. Methods The limbal tissues of 15 New Zealand white rabbits (2 mm ×2 mm) were treated with 1.2 U/ml lyase Ⅱ in a 5% CO2 incubator at 37℃ for 20 min, and then cultured on the epithelial surface of human amniotic membrane with infusion solution for 2 weeks, followed by air-liquid culture for 2 weeks. During the culture period, transmission electron microscopy was performed regularly every week to observe the formation process of junction complexes. After the 4th week of culture, the differentiated epithelial cells were identified by AE5 immunohistochemistry, periodate-Schiff, hematoxylin-eosin staining methods, and goblet cells were identified, and the morphological characteristics of the cells were observed. Results Rabbit limbal epithelial cells could fill the surface of human amniotic membrane for 10~14 days in vitro, The epithelial cells differentiated by AE5 immunohistochemical staining were positive, but the epithelial cells differentiated by periodate-Schiff staining were negative, and hematoxylin-eosin staining showed that there was no significant difference between the epithelial cells differentiated from rabbit limbal epithelial cells and normal rabbit corneal epithelial cells; No junction complex with amniotic basement membrane was formed from the culture to the 14th day; Conclusion the differentiated cells of rabbit limbal epithelial cells cultured on human amniotic membrane in vitro are corneal epithelial cells, which have the same morphology as normal rabbit corneal epithelial cells, and only form the initial junction complex with the basement membrane of amniotic membrane.

Objective to investigate the effects of hydrogen peroxide (H2O2) on the distribution and expression of plasma membrane microencapsulin (CP) and phosphorylated CP-1 in human lens epithelial cells. Methods Human lens epithelial cells (SRA01/04) were stimulated with H2O2 at different concentrations and at different time points. The distribution of CP and phosphorylated CP-1 was observed by laser confocal microscope and immunofluorescence microscope. The expression level of CP and phosphorylated CP-1 were observed by immunoblotting. Results The plasma membrane and cytoplasm of human lens epithelial cells contained abundant CP by laser confocal microscope and fluorescence microscope; When the cells were stimulated by H2O2, the distribution of CP in the cytoplasm increased; When the stimulation time reached 1 h, the cell membrane was destroyed, but the distribution of CP could still be observed. In addition, the stimulation of H2O2 could cause the phosphorylation of CP-1. Immunoblotting assay showed that with the prolongation of the action time of H2O2 and the increase of the stimulation concentration, the expression of CP in the plasma membrane and the total protein of cells was downregulated. Conclusion After stimulation of H2O2, CP redistributed and may destroy the structure of plasma membrane microcapsules (caveolae), resulting in the downregulation of CP expression in human lens epithelial cells.

The study of proteomics can not only explain the law of life activity, but also provide theoretical basis for the elucidation of the mechanism of occurrence and development of some diseases and the way to solve them. Through the analysis and comparison of proteomics in individual physiology or pathology, disease-specific protein molecules can be found, which can provide molecular markers for early diagnosis of diseases, and can also become molecular targets for new drug design.The application of proteomics in ophthalmology research is reviewed.

Anticardiolipin antibody belongs to one of autoantibodies, which is related to thrombosis, thrombocytopenia and spontaneous abortion.In recent years, foreign scholars have done a lot of research from different angles, which shows that thrombotic diseases are related to anticardiolipin antibody.Therefore, the author reviewed the relevant literature in recent years, and summarized the epidemiology, immunological pathogenesis, clinical manifestations, ocular characteristics, diagnostic basis, differential diagnosis, prevention and treatment of anticardiolipin antibody positive patients.

The 17th International Congress of Eye Research (XVII International Congress of Eye Research) was successfully held in Buenos Aires, Argentina from October 29 to November 3, 2006. The International Eye Research Congress is an International event hosted by The International Society for Eye Research, which aims to exchange and improve The level of International Research in The field of ophthalmology. Founded in 1968, the International Association for Eye Research held a symposium on lens research in the Netherlands in 1971 and the first international congress on eye research in Italy in 1974. After that, the association decided to hold a conference every two years and chose the address of the conference all over the world. It is currently the largest and representative international conference in the field of ophthalmology research.

Purtscher retinopathy refers to a special retinopathy that occurs after severe crush injury or comminuted fracture of thorax and abdomen. The fundus manifests as cotton wool patches around the optic head, bleeding, yellow-white retina patches and other changes. Purtscher-like retinopathy refers to the special manifestation of Purtscher retinopathy in the fundus of non-traumatic systemic diseases, such as systemic lupus erythematosus, acute pancreatitis, chronic renal failure and other diseases.

lactoferrin (LF) is the most important antibacterial protein in tears [1-2], which plays an active role in protecting the ocular surface tissue, and its content can reflect the exocrine function of lacrimal gland to a great extent. Dry eye is an ocular surface disease closely related to tear film, with complex etiology and diverse clinical manifestations, and it is difficult to diagnose.

In recent years, with the widespread application of antibiotics and glucocorticoids, fungal ophthalmopathy has increased year by year. In December 2005, a patient with severe fungal infection of both eyelids was diagnosed and treated in our hospital.

megalocomea is a rare abnormal development of the anterior segment of the eye, with congenital, bilateral, non-progressive and hereditary characteristics. It is generally considered that megalocomea is an independent disease. The transverse diameter of the cornea is more than 13 mm and the vertical diameter is more than 12 mm. The lens, the suspensory ligament of the lens and the ciliary ring are also enlarged accordingly, and the cornea itself is histologically normal. Clinically, its changes are limited to the anterior segment of the eye, while the intraocular pressure, fundus and visual function are all within the normal range.

Van der Hoeve syndrome is blue sclera-brittle bone (deafness) syndrome.Nevus of ota is congenital ocular skin melanocytosis nevus, also known as ota syndrome.It is a blue nevus surrounding the orbit, eyelid and eyebrow skin, accompanied by ipsilateral conjunctiva, sclera and uveal diffuse nevus.The genetic mode is mostly autosomal dominant inheritance, but there are also recessive ones [1]. The author diagnosed and treated a patient with this disease in August 2006.

Case 1: A 76-year-old female came to the emergency department of our hospital on July 18, 2002 due to swelling and pain in her left eye after being smashed by a clothes hanger, blurred vision, and nausea and vomiting for 6 h. The history of acute angle-closure glaucoma in both eyes was 13 years.

Pilocarpine is a post-holiday cholinergic drug, which has the effect of constricting pupils and lowering intraocular pressure, and is mainly used for the treatment of glaucoma.In 2003, a patient with immediate systemic adverse reactions was treated in our hospital due to the use of 2% pilocarpine ophthalmic solution. The report is as follows.

The patient was a 34-year-old female who came to the clinic due to upper respiratory tract infection, fever, pain in both eyes and headache (the symptoms were very similar to medical symptoms at the time of consultation, so the intraocular pressure was not measured). The patient had no history of ophthalmic diseases, trauma, systemic diseases such as hypertension and diabetes, and no family history of glaucoma.

The patient was a 39-year-old male who came to our hospital on November 9, 2006 due to visual field defect under the left eye and visual acuity decrease for 16 days. The patient had a history of myopia for more than 20 years, and had undergone laser surgery for binocular myopia in 2003. The specific surgical procedure is unknown.

The standard compatibility type question is one of the single-choice type B questions (type B1), in the form of five alternative answers and several questions, that is, several groups of examination questions are provided, each group of examination questions uses the five alternative answers listed before the examination questions together, and one answer closely related to the examination question is selected from them. Each alternative answer may be selected once, multiple times or not selected.

Led by Professor Ren Qiushi, chief scientist of the National Major Basic Research Program (973) and director of Shanghai Key Laboratory of Laser Medicine, the Institute of Laser and Photonic Biomedicine of Shanghai Jiaotong University is an institution specializing in the basic function principle and application of laser and optoelectronic technology in the biomedical field. The study takes the operating concept of Wellman Laboratories of Photo-medicine at Harvard University, Beckman Laser Institute and Medical Clinics at the University of California as the model, and combines teaching, scientific research, talent training and transformation of scientific research achievements. At present, the Institute has established cooperative relations with many internationally renowned universities, such as Wellman Joint Laboratory of Laser Medicine of Harvard University, Beckman Institute of Laser Medicine of University of California, and Medical Physics Research Center of Heidelberg University in Germany. The Institute belongs to the Department of Biomedical Engineering, School of Life Science and Technology, Shanghai Jiaotong University, which is a national key first-class discipline, doctoral program and postdoctoral mobile station.