Volume 40 ·  Issue 08 ·  2004

Uveitis is a common blinding eye disease. According to reports, about 10% of blindness in the United States is caused by uveitis, which occupies the 3rd to 10th place among blinding eye diseases in China [1,2]. Therefore, it is widely concerned by ophthalmologists. However, due to various reasons, there are still many problems in the diagnosis and treatment of uveitis, mainly manifested in the simplification of diagnosis, the formatting of treatment and the complexity of medication [3-5].

Objective to investigate the expression of Fas and FasL mRNA in peripheral blood lymphocytes of patients with Vogt-Koyanagi-Harada (VKH) syndrome. Methods From January to June 2000, the peripheral blood of 16 patients with VKH syndrome and 19 normal subjects was collected, the total RNA of lymphocytes was extracted, and the RNA was reverse transcribed into cDNA. Then, the whole process of Fas and FasL PCR amplification was detected by fluorescent quantitative polymerase chain reaction (FQ-PCR) in real time, and the expression of Fas and FasL mRNA in the samples was calculated. Results The expression of Fas mRNA in peripheral blood lymphocytes of patients with VKH syndrome was significantly higher [(1.6 ± 2.0) ×106] than that of normal controls [(5.7 ± 2.0) ×105] (t =4.50, P<0.05), and the expression of FasL mRNA [(1.8 ± 1.5) ×106] was also significantly higher than that of normal controls [(4.8 ± 3.5) ×105] (t =9.57, P<0.05). Conclusion VKH

Objective to investigate the clinical application value of laser protein cell detector for the quantitative determination of aqueous humor flash and inflammatory cells in patients with anterior uveitis. Methods the noncontact quantitative detection of aqueous humor flash and inflammatory cells in 110 patients with anterior uveitis (194 eyes) and 52 normal subjects (52 eyes) from December 2001 to April 2002 in Zhongshan Eye Center was performed by FC-2000 laser protein cell meter (1aserflare cell meter, LFCM). Before the detection, all patients were examined in detail under slit lamp microscope, and the aqueous humor flash grading and quantitative detection of cells were performed. Results The average values of O ~2 aqueous humor flash in patients with anterior uveitis detected by LFCM were 7.9, 29.5 and 189.O pc/ms, respectively; The results of grade 3~4 aqueous humor flashes showed warning or undetectable due to the large background interference. The slit lamp microscopy of grade 0~2 aqueous humor flashes in patients with anterior uveitis showed a positive correlation with the results of laser protein determination (r =0.75, P<0.001); The mean values of inflammatory cells in aqueous humor were 1.5, 12.1, 33.9, 84.9 and 193.1/0.5 mm3 in patients with uveitis before LFCM (grade 0-4 aqueous humor flash), respectively, and there was a positive correlation between slit lamp microscopy and laser protein cell count (r =0.72, P<0.001). Compared with normal human aqueous humor (0.9/0.5 mm3), the values of inflammatory cells were significantly higher (t =7.351, P<0.05). Conclusion LFCM can accurately judge mild and moderate blood aqueous humor barrier destruction and inflammatory response changes, and this examination is of great value for judging the inflammatory response of anterior segment of eye and guiding clinical medication. (Chinese Journal of Ophthalmology, 2004, 40: 510-513)

Objective to investigate the long-term efficacy of combined anterior and posterior segment surgery with temporary artificial cornea in the treatment of complex ophthalmopathies.Methods From June 1994 to June 2001, 107 patients (107 eyes) with complex anterior and posterior segment lesions underwent vitreoretinal surgery with temporary artificial cornea and penetrating keratoplasty in our hospital. After operation, glucocorticoid was applied locally and systemically, and the visual acuity, intraocular pressure, corneal graft and fundus condition of the patients were followed up. (2) retinal repositioning; Results 78 eyes (72.9%) reached the standard of surgical cure, 92 eyes (86.0%) had eyeball preservation after surgery, 34 eyes (31.8%) had graft immune rejection after surgery, 13 eyes (12.2%) had eyeball atrophy, and 15 eyes (14.0%) had secondary glaucoma. The cure rate of retinal surgery in eyes with proliferative vitreoretinopathy (PVR) before surgery was significantly different from that in eyes without PVR (x2=3.90, P<0.05). Conclusion Combined anterior and posterior segment surgery of temporary subcorneal artificial eyeball is an effective method for the treatment of obvious corneal opacity combined with vitreoretinopathy. The main reasons for long-term failure are endothelial decompensation of corneal graft and proliferative vitreoretinopathy. (Chinese Journal of Ophthalmology, 2004, 40: 514-516)

Objective to explore the feasibility of using tissue engineering technique to construct corneal stromal layer tissues in rabbits.Methods Forty New Zealand Dabai cells, namely, 20 pairs of female rabbits and their biological children, were isolated and obtained.The corneal stromal cells of newborn rabbits were isolated, expanded, cultured, confluenced, inoculated with polyhydroxyacetic acid (PGA) to form cell-biomaterial complexes.The corneal stromal cells were labeled with green fluorescent protein (GFP) to trace the process of corneal stromal layer.Meanwhile, the contralateral cornea was only transplanted with PGA as the material control.After 8 weeks, the samples were collected and histological sections were performed, and the collagen type I was detected by Western blot and the diameter distribution of collagen fibers was determined under electron microscope.Results 8 weeks after operation, the experimental cornea gradually returned to transparency, and neonatal corneal stromal-like tissues were formed. The collagen was parallel to the corneal surface and arranged neatly, and the histological structure was close to normal stromal tissues.Western blot detection in the experimental group showed that the stromal cells in the neonatal tissues expressed collagen type I; There was no significant difference between the diameter of collagen fiber [(29.0±4.7) nm] and the normal corneal stromal tissue [(28.5±3.5) nm] under electron microscope (P>0.05). There was no new stromal tissue formation in the normal cornea of the control group. GFP-labeled corneal stromal cells traced the corneal stroma at the 8th week, and the new tissue showed green color under fluorescence microscope, suggesting the expression of GFP. Conclusion Tissue engineering technique can be used to construct corneal stromal tissue in rabbit recipient cornea. (Chinese Journal of Lumbar Medicine, 2004, 40: 517-521)

Objective to investigate the expression of vascular endothelial growth factor (VEGF) and its FLK1 receptor in choroidal neovascularization (CNV) induced by Krypton laser in brown Norwegian (BN) rats. Methods Retinal photocoagulation was performed on 30 male BN rats. CNV model was established by Krypton laser. Fluorescein fundus angiography (FFA) was performed at 3, 7, 14, 21, 28 and 56 days after photocoagulation, respectively. After sacrificing the rats, the eyeballs were removed, and histopathological samples were made for observation. VEGF mRNA was detected by in situ hybridization, and VEGF and FLK1 receptor were detected by immunohistochemistry. Results VEGF mRNA was expressed in the retinal ganglion cell layer, inner nucleus layer, pigment epithelium layer, retinal and choroidal vascular endothelial cells of normal BN rats. The expression level of VEGF mRNA in retina decreased gradually from 3 to 21 days (P<0.01); There was no significant difference in the expression level of VEGF mRNA between 21 days and 28 days and 56 days (P>0.05). 7 days after photocoagulation, FFA examination showed disc-shaped fluorescein leakage in the photocoagulation area. Pathological sections showed CNV formation under the retina, and the expression of VEGF mRNA in CNV was positive; The positive staining area and absorbance (A) value of VEGF mRNA in CNV increased gradually from 7 to 21 days (P<0.01); There was no significant difference in the expression of VEGF mRNA after 21 days (P>0.05). The expression of FLK1 receptor was positive in normal retinal and choroidal vascular endothelial cells and retinal ganglion cell layer; 7 days after photocoagulation, the expression of FLK1 receptor in CNV was positive; With the proliferation of CNV, the positive staining area and A value of FLK1 receptor increased gradually (P<0.01); There was no significant difference in the expression level of FLK1 receptor on 21 days compared with 28 days and 56 days (P>0.05). Conclusion VEGF and FLK1 receptor accumulate in the injured areas of retina and choroid, and after effective binding, they induce the differentiation of endothelial cells and the formation of CNV under the retina. (Chinese Journal of Ophthalmology, 2004,40:522-527)

Objective to investigate the inhibitory effect of basic fibroblast growth factor (bFGF) antisense oligonucleotide and its liposomes on the proliferation of rat lens epithelial cells (LEC). Methods Rat LEC were cultured in vitro, and bFGF antisense and sense oligonucleotide and its liposomes were added respectively after cell passage. After 24 h culture with nutrient solution and drug-free liposomes as controls, the proliferation of the cells was measured by thiazole blue (MTT) assay, and the expression of bFGF mRNA was measured by reverse transcription polymerase chain reaction (RT-PCR). Results The cell growth of rat LEC was observed in the shape of polygonal shape after 24h primary culture, and the cells converged into pieces after 2~3 weeks culture; The absorbance (A value) of bFGF antisense oligonucleotide group (group I), sense oligonucleotide group (group II) and nutrient solution group (group V) were 0.138 ± 0.074, 0.325 ± 0.097 and 0.370 ± 0.077, respectively. The A value of group I was significantly lower than that of group II and V (P =0.024, 0.005). The A values of bFGF antisense oligonucleotide liposome group (group III), sense oligonucleotide liposome group (group IV) and drug-free liposome group (group VI) were 0.128 ± 0.032, 0.238 ± 0.120 and 0.348 ± 0.032, respectively. The A values of group III were significantly lower than those of group VI (P =0.000). The results of RT-PCR showed that the amounts of bFGF antisense oligonucleotide, bFGF sense oligonucleotide and amplification products of nutrient solution were 0.33, 0.99 and 0.85 μ g, respectively, with 2 μ g total RNA as template and 30 cycles; The amounts of amplification products of bFGF antisense oligonucleotide liposome, bFGF sense oligonucleotide liposome and drug-free liposome were 0.23, 0.48 and 0.56 μ g, respectively. Conclusion Rat LEC can be cultured in vitro; The bFGF antisense oligonucleotide and its liposome can decrease the mRNA content of bFGF and inhibit the proliferation of LEC in cultured rats in vitro. (Chinese Journal of Ophthalmology, 2004, 40: 528-532)

Objective to observe the effects of fetal bovine serum organ culture medium and human umbilical cord serum organ culture medium on the morphology, histology, ultrastructure, enzyme activity and metabolism of porcine corneal endothelial cells. Methods Organs were cultured in closed culture at 31℃ within 4 weeks and dehydrated for 24h. 113 pig eyes were selected, of which 100 corneas were divided into two groups for paired organ culture and preservation. Group 1 (50 corneas): culture medium I (containing 10% fetal bovine serum) was applied; In group 2 (50 corneas, contralateral corneas in group 1), 13 corneas were used as normal controls in culture medium Ⅱ (containing 10% human umbilical cord serum). Twelve corneas were taken out from each group every week for morphological analysis of endothelial cells, HE staining and enzyme histochemical staining. Scanning electron microscopy was performed at 2 and 4 weeks of preservation. The pH value, glucose and lactate concentrations of the culture medium before and after preservation were measured. Microbiological examination was performed during organ culture. Results During organ culture, the monolayer of corneal endothelial cells remained intact, and the pleomorphism increased. There was no significant difference in the density of corneal endothelial cells, the coefficient of variation of cell area and the proportion of hexagonal cells between the two groups within 4 weeks of preservation. Compared with the normal corneas of porcine preserved for 4 weeks, the mean endothelial cell loss rate was 10.98% in group 1 and 10.85% in group 2. There was no significant difference in histology, ultrastructure and enzyme activity between the two groups after corneal organ culture. Enzyme histochemical staining showed that the enzyme activity of epithelial and endothelial cells was strong, while the enzyme activity of stromal cells decreased with the prolongation of preservation time. The corneal metabolism state was good, and the contamination rate of organ culture fluid was 6%. Conclusion Both organ culture fluids can maintain the activity of corneal endothelium for 4 weeks, and it is speculated that human umbilical cord serum can replace fetal bovine serum as the component of corneal organ culture fluid. (Chinese Journal of Ophthalmology, 2004, 40: 533-538)

Objective to investigate the possibility and mechanism of endocytosis of macromolecular soluble substances in photoreceptor extracellular matrix (IPM) in retinal pigment epithelium (RPE) cells. Methods Serum albumin (F-FSA) after fluorescence labeled formaldehyde was used as an indicator of scavenger receptor ligand. The endocytosis of F-FSA in RPE cells was observed by fluorescence microscope and electron microscope. This effect increased with the increase of ligand concentration and increased with the prolongation of incubation time; Competition exists between the same or similar ligands; Conclusion RPE cells can take up negative-charged macromolecular substances through receptor-mediated endocytosis, and scavenger receptor may be the receptor mediating this effect; Receptor-mediated endocytosis of RPE cells may be an important mechanism of macromolecular metabolism in IPM. (Chinese Journal of Ophthalmology, 2004, 40:539-544)

Objective to investigate the effects of epidermal growth factor (EGF), interleukin-1 (IL-1), gamma interferon (IFN-γ) and somatostatin-8 peptide (SS-8 peptide) on the formation of opacity in posterior lens capsule and the mechanism of influencing collagen synthesis.Methods Cells were inoculated in 96-well culture plates and adhered to the wall. After adhering to the wall, cell culture medium containing 10-2-102ng/ml EGF, 10-105ng/ml IL-1, 10-11-10-7mol/L SS-8 peptide and 10-105U/ml IFN-γ was added respectively. The effect on collagen synthesis was detected by 3H-proline incorporation method. Results EGF 10-1~102ng/ml, IL-1 102~105ng/ml significantly promoted collagen synthesis, IFN-γ 103~105U/ml, SS-8 peptide 10-10~10-7mol/L significantly inhibited collagen synthesis; 1 ng/ml of EGF and 103 ng/ml of IL-1 can increase the expression of type Ⅰ and Ⅲ procollagen mRNA, while 103U/ml of IFN-γ and 10-9mol/L of SS-8 peptide can decrease the expression of type Ⅰ and Ⅲ procollagen mRNA. Conclusion EGF and IL-1 are involved in the collagen synthesis in the formation of posterior lens capsule opacity, and their effects on collagen synthesis are at the level of transcription; The effects of IFN-γ and SS-8 peptides suggest that they can be used to prevent and treat posterior lens capsule opacity. (Chinese Journal of Ophthalmology, 2004,40:545-548)

Objective to compare the iatrogenic injury caused by different microinjection techniques of retinal pigment epithelium (RPE) transplantation into the subretinal space of Lewis rats. Methods Forty-five eyes of Lewis rats were divided into three groups, and the subretinal space was injected with hTERT-RPE1 cells transfected with fluorescein protein. The first group was injected with Hamilton syringe, the second group was injected with Eppendorf microsampler, and the third group was injected with input pump syringe. The degree of iatrogenic injury in the host eyes was evaluated under light microscope. Results 12 eyes of the injection pump syringe group had no injury, while 14 eyes of the Eppendorf microsampler group and the Hamilton syringe group had severe or mild injury. Conclusion The low-speed motor pump of the injection pump syringe can provide a slow and controlled fluid injection flow, which can significantly reduce the iatrogenic injury to the host eyes when RPE is transplanted into the subretinal space. (Chinese Journal of Ophthalmology, 2004, 40: 549-551)

In 1979, Sugitani et al. first reported the existence of about 2.00D accommodation in the surgical eye after intraocular lens implantation, and called it pseudo-accommodation force. Since 2001, we have conducted relevant research on pseudo-accommodation force in the surgical eye after phacoemulsification and folding intraocular lens implantation, and the results are reported as follows.

As the main structural protein of the lens, α-lens protein has molecular bridesmaid characteristics, which is of great significance for maintaining the transparency of the lens for a long time [1,2]. In order to investigate its mechanism, we compared and observed the inhibitory effect of α-lens protein on catalase thermal aggregation in the cortex and nucleus of the clear lens, the lens of patients with different ages and the lens with different degrees of opacity in age-related cataract.

Recent studies have found that nitric oxide (NO) is one of the important substances causing oxidative damage of cells and is related to many kinds of ocular diseases.In this study, in vitro lens culture technique was used to detect the changes of NO and nitric oxide synthase (NOS) contents in the lens during the formation of cataract, to explore the relationship between nitric oxide (NO) and cataract formation, and to observe the inhibitory effect of taurine.

tetramethylpyrazine is one of the active ingredients extracted from Ligusticum chuanxiong, also known as tetramethylpyrazine, which can be synthesized artificially and is widely used in clinic.It is mainly used for the treatment of cerebral thrombosis in occlusive cerebrovascular disease [1], and has good curative effect on improving visual function of glaucoma patients, simple diabetic retinopathy and retinal vein occlusion in ophthalmology [2-4]. In this study, tetramethylpyrazine hydrochloride was injected intraperitoneally to investigate its pharmacokinetic changes in the vitreous body of rabbits. The results are reported as follows.

Diabetic retinopathy is an important manifestation of diabetic microvascular disease and one of the serious complications of diabetes, while hyperglycemia is an important risk factor for the occurrence and development of diabetic microvascular complications [1-3].

transpupillary thermotherapy (TTT) is a kind of hyperthermia technique for the treatment of local diseases.The preliminary clinical results show that TTT has the advantages of high efficiency and small tissue damage in the treatment of choroidal melanoma, retinoblastoma, choroidal hemangioma and choroidal neovascularization (CNV).

The patient was a 70-year-old female who came to our hospital in December 2001 because of blurred vision in her right eye for 3 months. The patient had undergone phacoemulsification and intraocular lens (IOL) implantation in the left and right eyes in October and December 2000 because of uveitis, refractive error and complicated cataract in both eyes, respectively. During the operation, the right eye was implanted with hydrogel Model H60M (product serial number: SN: 5WJ239) folded IOL from Bausch & Lomb.

Uveitis is a common type of autoimmune eye disease. At present, there are about 3~4 million patients with uveitis in China, accounting for the 3rd ~10th blind eye disease [1]. The patients with blindness caused by uveitis in the United States account for about 10% ~15% of the total blind people [2]. Because the diagnosis of uveitis is mainly based on clinical manifestations, there is a lack of specific etiological examination methods; Therefore, ophthalmologists often feel very difficult when diagnosing and differentiating diseases and selecting relevant laboratory examination items [3-5]. In view of this, we expound and analyze the commonly used laboratory examination items and their indications.