Objective To investigate the retinal toxicity and verify the safe dose of intravitreal injecting fluconazole. Methods Twelve healthy adult white rabbits were divided at random into 6 groups：a normal control group and 5 groups received intravitreal injection of a single dose of fluconazole ranging from 10 to 200 mu;g respectively.Retinal toxicity was examined by ophthalmoscopy, electroretinography, light and transmission electron microscopy (TEM) on the third and fourteenth day after injection. Results The ultrastructures of the retinal tissues of the normal control group and fluconazole 10～150 mu;g groups were normal on the third and fourteen day after injection.The light microscopy and TEM showed that cells of all the retinal layers in the 200 mu;g group revealed apparent degenerative changes on the fourteenth day after injection, and the light microscopic picture showed the vacuolar degeneration of outer segments of photoreceptors, the nuclei in outer nuclear layer drop out into inner segments, the vacuolar degeneration of nerve fiber layer, and the proliferation of pigment epithelium. TEM revealed expansion of paranucl eus space and karyopyknosis of the bipolar cells, the swelling of nerve fibers and disappearance of the synapses in the inner plexiform layer, the vacuolation and disappearance of microvilli of the pigment epithelium cells. Conclusion The safe dose of fluconazole injected intravitreally should be 100～150 mu;g. (Chin J Ocul Fundus Dis，2000，16：139-212)
OBJECTIVE:To verify the safe dose of cephradine in intravitreal injection. METHODS:After injecting different doses of cephradine(100mu;g,200mu;g,250mu;g,300mu;g,400mu;g)into vitreous cavity of different group of rabbits the activities of the retinal enzymes (SDH,LDH )on different time (Id,3d, 7d ) were determined respectively, and the histological and ultrastructural changes of retinas were also observed simuhaneously. RESULTS:The activity of rellnal SDH and LDH was found to be decreased gradually with tbe icreasing of the dosage of intravitreal cephradine. The activities of SDH and LDH were found in the lowest level on tile 3rd and lsl day,but they recover to normal levels on tile 7th day after intravitreal in}eetion in 100mu;g,200mu;g groups,and still lower tban normal in the other groups. Histologically,retinal edema was found both in 100mu;g and 200mu;g groups,but degradation of retinal cells,and loss of cones and rods were round in the 250mu;g, 300mu;g and 400mu;g groups. CONCLUSION: The safe dose of intravitreal injection of cepbradlnc is 200mu;g. （Chin J Ocul Fundus Dis,1997,13:139-142 ）
Objective To investigate the retinal toxicity and verify the safe dose of intravitreal injection of nonsteroid anti-inflamatory drug,diclofenac sodium.Methods Twenty-eight healthy adult white rabbits were divided at random into 7 groups and received in every right eye the intravitreal injection of a single dose of diclofenac sodium solution ranging from 0.4-0.1 mg/0.1ml respectively ,the left eyes were regarded as conreol ones.Before injection and on the 1st,3rd,7th,14th,21st,and 28th day after injection the electroretinography on both eyes was examined.On the 28th day after injection the retinas of two rabbits of every group were examined by using light microscopy.On the 10th and 30th day after injection the retinal tissues around the optic nerve sisk of two eyes from every group at random were tested by using transmission electron microscopy.Results The retio of amplitude ofb wave of electroretinography in 0.4mg and 0.5mg groups had no sighnificant difference from groups before injection,the retinal tissues showed no structural changes in light and ecectron microscopy examination.The ratio of amplitude ofb wave of photoptic electroretinogrphy in 0.6mg groups in the early stage after injection was markedly reduced（P＜0.05）and returned to that before injection with time,reversible change of the edematou retina was discovered.The ratio of amplitude of b wave of electroretinography in 0.7-1.0mg groups was distinctly descreaded after injection（P＜0.05 or P＜0.01）,the cells of all the retinal layer revealed apparent and irreversible damage.Conclusion The largest dose of safety of intravitreal diclofenac sodium should be not more than 0.6mg.The toxic effect of intravitreal diclofenac sodium on retina is concerned mostly to cones and rods.
Objective To investigate the effects of gamma;-interferon (IFN-gamma; on the expression of costimulatory molecules B7-1 (CD80) and B7-2 (CD86) and Fas/FasL in human retina. Methods Nine human eyes were obtained from the eye-bank of Zhongshan Ophthalmic Center. Six eyes were used for making retinal wholemounts, and 12 retinal wholemounts from each eye were put into the 24-hole culture board which had 2ml DMEM/F12 culture medium in each hole. The wholemounts were divided into 3 groups whose concentration of IFN-gamma; was 0, 200, and 1000 U/ml respectively. After cultured in 37℃ culture box (95%O 2，5%CO 2) for 24 hours, the expressions of B7-1 (CD80), B7-2 (CD86), and Fas/FasL on these retinal wholemounts were detected by immunohistochemical method. The retinal wholemounts from 3 healthy people were detected by immunohistochemical method as the control. Results Expression of FasL but not B7-1, B7-2 or Fas was found in the control group, while the expression of B7-1, B7-2 and Fas and increased expression of FasL were found after cultured with IFN-gamma;. Conclusion IFN-gamma; may be involved in the occurrence of ocular immune response and induction of apoptosis via the stimulation of expression of costimulatory molecules and Fas/FasL, which plays an important role in the activation of T lymphocytes. (Chin J Ocul Fundus Dis, 2006, 22: 117-119)
Objective To investigate the correlation of ascorbic acid distribution and retinal susceptibility to iron toxicity of the retina.Methods Autoclaved iron particles of 5 mg and 15 mg were implanted into the vitreous cavities of 32 Spragu-Dawley (SD) rats and 9 rabbits, respectively. The retinal sections of rats and rabbits were examined after hemotoxylin-eosin (HE) staining. Apoptos is of rabbits′retinal neurons was investigated by TdT-mediated dUTP-biotin nick-end labeling (TUNEL). Chinoy′s method was used to observe the distribution of as corbic acid in the retinae of the 2 kinds of animals.Results In rats, histological and structural densification was observed only in the photoreceptor cells after implantation of the iron particles. In rabbits, however, histological and structural destruction as well as TUNEL-positive nuclei were observed in all neuronal layers of the retina 3 days after the implantation of the iron particles. Silver granules reduced by ascorbic acid from silver nitrate were observed only in the outer nuclear layer in normal rats retinae, while they were observed evenly throu ghout all layers of rabbits′retinae. Conclusions The suscept ibility of retina to iron toxicity is correlated to the distribution of ascorbic acid in retina. (Chin J Ocul Fundus Dis，2003，19：269-332)
Objective To inverstingate the effect of perfluorohexyloctane(F6H8)to the retina of rabbit eyes. Methods Fifteen vitrectomized New Zealand white rabbits were injectedF6H8(experiment group,12 rabbits ) and BSS(control group,3 rabbits) into vitreous cavity.Slit-lamp biomicroscopy and indirect ophthalmoscopy were performed pre- and postoperatively in all the eyes.Histopathological examination was done after the rabbits were sacrificed at the end of the study. Results A large clear balb was formed after intravitreal injection of theF6H8 in the vitreous was injected and no retinal detachment and cataract were found.The OPL was edematous and then thinned out in 4th week in experimental group.Degenerating cells was found in inner and outer nuclear layers.Cellular vaculoar degeneration was present in TEM. ConclusionF6H8 in vitreous cavity may cause significant side effects on retina,we could not recommend it to be used as an intraocular temponade.
ObjectiveTo evaluate the security of intravitreal injection with ciproflaxacin to retina.MethodsTweenty-four rabbits were randomly divided into 4 groups with 6 rabbits in each group. 0.1 ml ciproflaxacin in doses of 2 500,5 000,and 10 000 μg was intravitreally injected into the rabbits eyes, retrospectively. And 0.1 ml saline solution was injected into the vitreous body of the rats in the control group. Indirect microscope, light microscope and electroretinogram (ERG) were used to observe the changes of ocular fundus.ResultsNormal results of light microscopy and ultrastructure were found in 250 μg and 500 μg groups; irregularly arranged outer and inner nuclear layers, dropsical or even lost ganglion cells, and ultrastructural changes were in 1 000 μg group. There was no apparent difference of ERG′s a and b amplitudes before and after intravitreal injection with ciproflaxacin in each group.ConclusionIntravitreal injection with ciproflaxacin is safe, and 500 μg or less is the secure dosage in rabbits' eyes. (Chin J Ocul Fundus Dis, 2005,21:180-182)
Objective To observe the effects of high concentr at ion glucose on the calcium-activated potassium channel of rabbits′ retinal Müller cells. Methods The rabbits′retinal Müller cells were cultured in vitro under the condition of high concentration glucose, and identified by immunohistochemical staining and transmission electron microscopy. Patch-clamp technique was used to observe the changes of the calcium-activated potassium channel of retinal Müller cells caused by high concentration glucose at different time.Results High concentration glucose could inhibit the calcium-activated potassium channel of cultured retinal Müller cells in a time-dependent manner. Conclusion High concentration glucose may reduce the biological functions of Müller cells by inhibiting calcium-activated potassium channel. (Chin J Ocul Fundus Dis，2003，19：164-167)
Objective To investigate the effect of tetrandrine (Tet) on experimental choroidal neovascularization and the effect of Tet on retinal structure and function. Methods Choroidal neovascularization was induced in 20 Brown Norway (BN) rats (40 eyes) by diode laser (wavelength: 810 nm; exposal time: 0.1 second; facular diameter:100 mu;m; energy: 120 mW), and the rats were divided randomly into experimental and control group with 10 rats (20 eyes) in each group. In experimental group, 0.05 ml Tet with the concentration of 3.21 mu;mol/L was injected intravitreously 0 and 3 days after laser photocoagulation; in the control group, the rats underwent an intravitreous injection with the same volume of sodium chloride solution. The incidence of CNV was evaluated by fundus fluorescein angiography (FFA) 14 days after laser photocoagulation. Five right eyes of another Five healthy BN rats underwent intravitreous injection with 0.05 ml Tet with the concentration of 3.21 mu;mol/L, and an intravitreous injection with the same volume of sodium chloride solution was performed on the left eyes. Before injection, 1 hour, and 1 day after the first injection, and 1 hour, 1 day, 7 days, 14 days after e second injection the electroretinography (ERG) was performed on these 5 rats; 14 days after the second injection, the retinae were examined by light microscopy and transmission electron microscopy. Results The incidence of CNV was 23.26% in experimental group，which was obviously lower than that in the control group (63.33%) (Plt;0.01). The ratio of amplitude of b wave of ERG in the rats undergone intravitreous injection with 3.21 mg/ml Tet didnprime;t differ much from which before the injection (Pgt;0.05). There were no structural changes of retinal tissues examined by light and electron microscopy. Conclusion Tet may inhibit choroidal neovascularization in rats; there isnprime;t any significant toxic effect of intravitreous injection with Tet on retina at the dosage of 3.21 mu;mol/L. (Chin J Ocul Fundus Dis, 2006, 22: 242-244)
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.