PURPOSE: To investigate the treatment of severe bacterial endophthalmitis. METHODS:The curative effects of vitrectomy after intravitreal antibiotics and steroids (IVAS)for the treatment of 23 patients with bacterial endophthalmitis
(group I)and vitrectomy and IVA at the same time for the treatment of 28 patients with bacterial endopbthalmitis (group I)were analyzed retrospectively. RESULTS: The rate of curative effects of two groups were similar,while the marked curative effects in group I (47.8% )was significantly higher than that of the group I (17.9%). The average period of eliminating infiamation of group I was longer than that of group I , and the incidence of postoperative retinal detachment of group Ⅱ was 3 times more than that of group I . CONCLUSION :It was indicated that vitrectomy after IVAS may increase the security of vitrectomy and the curative effects of severe bacterial ndophthalmitis.
Objective To observe the regulation effect of transforming growth factor alpha (TGFalpha;) on expression of glutamate transporter（GLAST）and ingestion activity of retinal Muuml;ller cells in mice. Methods To take the retinal tissue of Kunming mouse at postnatal 7～10 day, and then cultured Muuml;ller cells according to literature. The 3～4 generation cultured cells of the same primary cell were divided into two groups at random: ① TGFalpha; group: maintained in different concentrations of TGFalpha; as 50, 75, 125 and 150 ng/ml, 3 holes in each concentration;② Control group: cultured by Eagle culture medium which improved from Dulbeccon and contained 20% fetal calf serum. The influence of different concentrations TGFalpha; on GLAST activity in Muuml;ller cells were observed by L-3H-glutamate uptake detection; the expression of GLAST mRNA in Muuml;ller cells was determined by RT-PCR; the expression of GLAST protein was detected with immunocytochemical staining. Results With the increase of TGFalpha; concentration, both L3H glutamate uptake and GLAST mRNA expression were increased. The L-3H-glutamate accumulation had got to the maximum uptake at concentration of 125 ng/ml, which was 266% of that in control group, meanwhile, the expressions of GLAST mRNA also got to the maximum as 4 times of control group. Immunocytochemical staining indicated that the effect of 125ng/ml TGFalpha; on expression of GLAST protein was higher than that in the control group, the differences between two groups were statistically significant (Plt;0.05). Conclusion TGF-alpha; can increase GLAST activity through up-regulating the expression of GLAST mRNA and protein.
Objective
To observe the effects on rabbit corneas and retinas after single intravitreal injection of voriconazole at different doses.
Methods
According to the randomization table, 25 healthy rabbits were randomly divided into control group, and voriconazole 50, 100, 200, and 400 μg groups. Therefore, there were 5 rabbits in each group. The eyes of control group received intravitreal injection of 0.1 ml balanced saline solution, and those treatment groups received 0.1 ml voriconazole injection of corresponding dose. Before the injection and 1, 7, and 14 days after the injection, endothelial cell counts and corneal thicknesses were measured; full-field electroretinogram were performed and b-wave amplitudes in maximal combined reaction (Max-R) were recorded. On 14 days after the injection, histologic structures were observed by light microscope and transmission electron microscope.
Results
There was no significant difference in endothelial cell counts (F=0.320, 0.291, 0.467, 0.649) and corneal thicknesses (F=0.214, 0.284, 0.360, 0.225) with those of control group at any time points (P > 0.05). Before and 1 day after the injection, b-wave amplitudes of each voriconazole group had no significant difference compared with those of control group (F=0.220, 0.106; P > 0.05). On 7 days after the injection, b-wave amplitudes decreased significantly at doses of 200 μg and 400 μg (P < 0.05). On 14 days after the injection, there was no significant difference between the the amplitude of 200 μg group and that of control group (P > 0.05). However, the amplitude of the 400 μg group decreased continuously and there was still significant difference (P < 0.05). Light microscopy did not reveal any corneal abnormality in both control group and voriconazole groups. The retinas were normal except that of the 400 μg group, which hadathinner and degenerated inner nuclear layer and disordered photoreceptor layer. Under transmission electron microscope, there were no ultrastructure damages of corneas in both control group and voriconazole groups, either. The rabbit retinas of the 50 μg and 200 μg group have normal inner nuclear layer and photoreceptor layer, but degrees of changes in both layers were observed in the eyes of 200 μg and 400 μg group.
Conclusions
There is no obvious effects on rabbit corneas and retinas after single intravitreal injection of voriconazole at he dose less than or equal 100 μg. There are no obvious effects on rabbit corneas at the dose of 200 μg and 400 μg, while there are damages to the retinas in both functions and histological structures.
Objective To observe the efficacy of the anti-tumor necrosis factor-alpha; monoclonal antibody (TNF-alpha; MCAb) in the treatment of experimental autoimmune uveoretinitis (EAU). Methods EAU animal models were induced by interphotoreceptor retinoid-binding protein (IRBP) R16 peptide with immunization. The rats were divided into 2 groups according to the injection times. TNF-alpha; MCAb was administered intravenously on day 6 or 4, 6 and 8 post-immunization respectively, and then to observe the clinical expression by slit-lamp microscope. Meanwhile, take the rats which did not accept TNF-alpha; MCAb as control group. Delayed type hypersensitivity (DTH) responses were measured on day 13 post-immunization of IRBP R16; the rats were killed on day 14 post-immunization of IRBP R16, and then enucleated the eyes for histopathological examination. To detect the cytokine level of IFN-gamma;, IL-4 in serum and IFN-gamma; in aqueous humor by enzyme-liked immunosorbent assay (ELISA) on day 14 post-injection. The hyperplasia responses of antigen specific lymphocyte of draining lymph node cells were detected. Results The TNF-alpha; MCAb group had mitigated ocular inflammation and decreased pathological grades compared with the control group; the IFN-gamma; concentrations in aqueous humor and serum were decreased, IL-4 was increased in serum; DTH responses were decreased; the hyperplasia responses of draining lymphocytes to IRBP R16 peptide were decreased, all the differences were statistically significant (P＜0.01). The rats accepted TNF-alpha; MCAb thrice had much better curative effect than the rats injected once (P＜0.05). Conclusions Injection of TNF-alpha; MCAb can inhibit ocular inflammation and specific immune cells of EAU remarkably and change the Th1/Th2 balance. Many times injections of TNF-alpha; MCAb were more effective than once.
Objective To observe the therapeutic effects of ganciclovir (GCV) with different injection methods on experimental acute retinal necrosis (ARN). Methods The right eyes of 41 pigmented rabbits were infected by herpes simplex virus (HSV-1) (COS strain) to establish ARN animal model. After 24 and 72 hours, GCV was given by intravitreal injection (10 eyes), intravenous injection (11 eyes) and the intravitreal+intravenous injection (10 eyes); intravitreal injection of GCV and dexamethasone (6 eyes) was also included. Four eyes were not treated as the control. The dosage of GCV in intravitreal and intravenous injection was 800mu;g and 5mg/kg weight, respectively. Retina necrosis was observed and the grade was recorded 1-21 days after injection according to the grade standard of retinopathy. The maximum grades of retinal necrosis in different groups were compared. Results The grade of retinal necosis was 3.8 in the control group, and 0.2, 0.4, 0.8, and 2.2 in intravitreal injection, intravitreal+intravenous injection, intravitreal injection with GCV and dexamethasone, and intravenous injection, respectively, 24 hours after the model was set up. The effects of the first 3 groups were obviously better than the last group (P=0.003, 0.011, 0.045); while the difference among the first 3 groups were not significant (P=0.881、0.054、0.107). Seventy-two hours after the model was set up, the grades of retinal necrosis were above 1.4 in 4 groups, and the differences among the 4 groups were not apparent (P=0.214). Conclusions In the animal model of ARN, intravitreal injection with GCV can effectively decrease the grade of retinal necrosis. The difference among intravitreal injection, intravitreal+intravenous injection, intravitreal injection with GCV and dexamethasone, and intravenous injection is not significant.
Objective To observe the influence of cisplan on the expression of B7-H1 in retinoblastoma (RB) cells，and to investigate its mechanism. Methods Human RB cell line HXO-Rb44 cells were treated by 6 different concentrations of cisplan (0.000, 0.375, 0.750, 1.500, 3.000, 6.000 mu;g/ml), and their B7-H1 mRNA expression was determined by the reversetranscription polymerase chain reaction (RT-PCR) and fluorescence quantitative PCR (FQ-PCR); the B7-H1 protein expression was determined by immunofluorescence and flow cytometry. HXO-Rb44 cells were treated by 1.5 mu;g/ml cisplan for 0, 15, 30, 60, 120 min, then the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) was detected by Western blot.Results The expression of B7-H1 mRNA and protein in the 0.375, 0.750, 1.500, 3.000, 6.000 mu;g/ml group were significantly higher than that of the blank control group (F=395.478，112.03; P=0.000）. Western blot showed that cisplan (1.5 mu;g/ml) could activate ERK1/2 by increasing its phosphorylation in HXO-Rb44 cells. After cisplan treatment, the phosphorylation of ERK1/2 increased gradually and reached its peak at 30 min, and then went down gradually.Conclusion Cisplan can promote the expression of B7-H1 and activate ERK1/2 in RB cells.
Objective To observe the effect of medicineinduced posterior vitreous detachment (PVD) on proliferative vitreoretinopathy (PVR). Methods PVR was induced in the left eyes of 24 pigmented rabbits by intravitreal injection with platelet rich plasma. The rabbits were randomly divided into two experimental groups (group A and B) and one control group with 8 eyes in each group. Three hours later, the eyes in group A and B and the control group underwent intravireal injection with 1 U plasmin 0.05 ml+20 U hyaluronidase 0.05 ml, plasmin 0.1 ml, and balance salt solution 0.1 ml, respectively. The grade of PVR was recorded 1, 7, and 28 days after the intravitreal injection, and the eyes were examined by flash electroretinogram (FERG), B-scan, and retinal histopathological examination. Results The PVR models of rabbit eyes were induced successfully. On the 7th day after injection, complete and partial PVD was found in 5 and 3 eyes respectively in group A; partial PVD in 5 eyes and no complete PVD was observed in group B; there was no PVD in the other 3 eyes in group B and also in the eyes in the control group. On the 28th day after intravitreal injection, PVR grade of group A and B were both obviously lower than that of the control group（D=75.6, 98.9；P=0.003，P=0.011）; On the 7th and 28th day after injection, the b-wave amplitude in group A and B was significantly higher than that in the control group; PVR grade of the PVD eyes was lower than that of nonPVD eyes; PVR grade of the complete PVD eyes was only 0～1. Conclusions Three hours after the PVR models of rabbit eyes were induced, complete PVD induced by intravitreal injection of plasmin combined with hyaluronidase could prevent the development of PVR of rabbit eyes in some degree; partial PVD induced by plasmin alone or combined with hyaluronidase could relieve the development of PVR.
Objective
To investigate the effects of QUE on proliferation and DNA synthesis of cultured retinal pigment epithelium(RPE) cells with or without EGF.
Methods
With or without EGF, cultured RPE cells were treated with QUE by various concentrations(200,100,50,1mu;mol/L) and with QUE 200mu;mol/L at different times(24-168 hr), cells proliferation and DNA synthesis were evaluated by cell count method and the uptake of thymidine. The viability of cells was determined by trypanblue exclusion.
Results
The best concentration of QUE which inhibits proliferation and DNA synthesis of PRE cells was 200mu;mol/L. The significant inhibition effect of QUE occurred at 48hr, and the best inhibition of QUE occurred at 96hr. QUE had more powerful effect of antiproliferation on RPE cells, and the viability of RPE cells was over85%.
Conclusion
The results suggested that QUE could inhibit the proliferation of RPE cells in a dose-dependent and time-dependent manner, especially inhibit the proliferation induced by EGF stimulating. QUE had no cyto-toxic effect on RPE cells cultured in vitro.
(Chin J Ocul Fundus Dis,1999,15:27-29)
Objective To evaluate the safety repeated intravitreal injection of bevacizumab (Avastin) with different dosage in rabbitsprime;eyes. Methods Fourteen chinchilla rabbits were randomly divided into 3 groups, including both eyes of 2 rabbits in the control group,the right eyes of the other 12 rabbits in the experimental group,and the left eyes of the 12 rabbits in the experimental control group. The eyes in the experimental group underwent intravitreal injection of bevacizumab with the dosage of 2.5 mg/0.1 ml and 5.0 mg/0.2 ml of bevacizumab; the eyes in the experimental control group underwent intravitreal injection of normal saline with the same dosage as in the experimental group. Injections were performed every two weeks and lasted six weeks. Clinical observation and retinal function tests were performed before and two days after every injection. The eyes were sacrificed 1 week and 4 weeks after last intravitreal injection respectively.Electron and optical microscope and TUNEL were performed.Results After intravitreal injection,no obvious anterior chamber flare, abnormal change of the ocular fundus, or vitreous opacity and hemorrhage was observed in all of the eyes.No change was found by indirect ophthalmoscope,Bultrasonic inspection, ultrasound biomicroscopy and optical coherence tomography. The number of anterior chamber flare before and after the injection with the dosage of 2.5 and 5.0 mg, the difference among the 3 groups didnprime;t differ much from each other (Pgt;0.05).Amplitude and pattern of ERG responses and flash VEP were similar between the control and experimental groups (Pgt;0.05). Some inflammatory cells were found in the some bevacizumabinjected eyes 1 week after injection, and vanished 3 weeks later. The histological configuration of the retina didnprime;t change in both experimental control and the control group. Electron microscopy showed that plasma cells were presented and vacuolelike change was observed in part of the photoreceptor cells in 5.0 mg experimental group 1 week after injections.Cellular apoptosis was observed in the photoreceptor cell layer. The number of apoptotic cells was more in 5.0 mg experimental group than that in the control and experimental control group 1 week after injections (Plt;0.01). Conclusion Multiintravitreal injection with 5.0 mg bevacizumab may have mild toxicity to the retina in the rabbits.
Objective To investigate the therapeutic effects of throm bolytic drug infusion via carotid artery on experimental central retinal artery occlusion (CRAO), and observe the changes of fibrinolytic activity in the system ic circulation. Methods To dissolve the thrombi in 15 cats (30 eyes) with CRAO established by laser irradiating a branch of central retinal a rtery after intravenous injection of photochemical drugs, urokinase (UK) was dir ectly infused via carotid artery in 5 cats (10 eyes) in group A or intravenously injected in 5 cats (10 eyes) in group B, and isotonic saline solution was intra venously injected in 5 cats (10 eyes) in group C respectively. The patency of the artery was evaluated by fundus fluorescein angiography. Moreover, the changes of fibrinolitic activity in the blood were observed by blood biochemical examination. Results Four hours after UK infusion, the complete repatency proportion was 80% (5 cats 8 eyes) in group A, and 50% (4 cats 5 eyes) in group B. There was significant difference between the two groups. Besides, after the infusion, the indexes of coagulation, fibrinolysis, and anti-fibrinolysis in group A were better than those in group B and C (Plt;0.01). Conclusion In the treatment of experimental CRAO, thrombolytic drug infusion via carotid artery is better and more effective than via intravenous injection, which may provide a new method of thrombolytic drug delivery and animal models. (Chin J Ocul Fundus Dis，2004，20：186-188)
