Objective To observe the expression of cyclin dependent kinase 5 (Cdk5) and p25 in the pathogenesis of retinitis pigmentosa (RP) in Royal College of Surgeon (RCS) rats and its relationships with apoptosis. To explore the mechanism of Cdk5 and p25 induced photoreceptor apoptosis in the pathogenesis of RP. Methods Retinas of RCS and RCS-rdy+ rats were obtained at the ages of postnatal day 17, 25, 35, 60. The retinal structure and thickness of outer nuclear layer were measured by optical microscopy. The expression of Cdk5, p25, cleave-caspase 3 in the retina was evaluated by immunohistochemistry. The protein expression of cleave-caspase 3 in the retina was determined by Western blot. The apoptosis of retinal cells was examined by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL). The mean absorbance value of apoptotic cells was analyzed by SPSS 17.0 software. Results The retinal thickness of the RCS rats was significantly reduced in comparison to the RCS-rdy+ rats as the postnatal days progressed, particularly in the layer of rods and cones and the outer nuclear layer. The expression level of Cdk5, p25, cleave-caspase 3 of RCS rats increased from postnatal 17 days to postnatal 35 days, while decreased on postnatal 60 days; but there was no obvious change of above indexes in RCS-rdy+ rats. The protein expression of cleave-caspase 3 in the RCS rats was significantly increased with progression of postnatal days to postnatal 35; but there was no obvious similar change in RCS-rdy+ rats. The results of TUNEL showed that the apoptotic cells significantly increased in the outer nuclear layer of RCS rats from postnatal 17 days to postnatal 35 days, while decreased on postnatal 60 days; but there was no obvious change of above index in RCS-rdy+ rats. This study showed that there were significant correlations between the following variables: Cdk5 expression and p25 expression, Cdk5 expression and cleave-caspase 3 expression, Cdk5 expression and apoptotic cells, p25 expression and cleave-caspase 3 expression, p25 expression and apoptotic cells, cleave-caspase 3 expression and apoptotic cells. The partial correlation coefficients were 0.949, 0.808, 0.959, 0.887, 0.979, 0.852, respectively and the P value was 0.000. Conclusions The apoptotic cells significantly increases and the expression level of Cdk5, p25, cleave-caspase 3 of RCS rats increases from postnatal 17 days to postnatal 35 days. The tendency of apoptotic cells to increase is consistent with the change of Cdk5, p25, cleave-caspase 3 expression. The apoptosis of photoreceptor cells is related to increasing expression of Cdk5 and p25 in RCS rats. Cdk5 may be involved in the development of RP in RCS rats.
Objective To observe the characteristics of fundus autofluorescence (AF) in short wavelength AF (SW-AF) and Near Infrared AF (NIR-AF), and their relationship with visual fields. Methods Twelve patients (24 eyes) with primary RP were enrolled in this study. The patients included nine males (18 eyes) and three females (six eyes). The patients aged from 15 to 69 years, with a mean age of (35.33plusmn;15.03) years. All the patients were examined for color photography, SW-AF, NIR-AF, visual fields and optical coherence tomography examination. Results There were hyper-AF ring of varying sizes in posterior pole by SW-AF and NIR-AF examinations. The area of hypo-AF which located in SW-AF hyper-AF ring had a positive correlation with the area of hyper-AF in the NIR-AF (r=0.662，P＜0.05). OCT showed that outside the hyper-AF ring, there were disconnected inner segment/outer segment (IS/OS) junction and external limiting membrane, and thinned outer nuclear layer and retinal pigment epithelium. Peripheral retinal osteocytes-like pigmentation showed non fluorescence in SW-AF and NIR-AF. The plaque-like area showed mottled and low fluorescence examined by SW-AF. SW-AF hyper-AF ring had a positive correlation with visual fields (r=0.492，P＜0.05). Conclusions The area of hypo-AF inside of the SW-AF hyper-AF ring is related to visual fields in RP patients. The retinal structures in the hypo-AF area inside of the SW-AF hyper-AF ring, and in the NIR-AF hyper-AF region are normal.
Objective To observe the fundus characteristics of acquired immune deficiency syndrome （AIDS) with cytomegalovirus retinitis （CMVR）. Methods Twenty-seven AIDS patients （44 eyes）with CMVR were studied. All the patients had undergone the examinations of visual acuity, intraocular pressure, slit lamp microscope, indirect ophthalmoscope and color fundus photography. The fundus lesions were divided into active lesions and chronic lesions, and the active lesions were subdivided into central, peripheral and mixed types which involving both the posterior and peripheral fundus. Results Of 27 patients （44 eyes）, 19 patients（29 eyes）had active lesions. Five patients （six eyes, 13.6%） had central lesions （exudation， hemorrhage and vascular sheath in the posterior retina）, nine patients （15 eyes, 34.1%） had peripheral yellow and white granular lesions. Five patients （eight eyes, 18.2%） had mixed lesions. Chronic lesions were found in eight patients（15 eyes, 34.1%）, which showed pigment and scarring lesions along vascular branches. Conclusion The fundus lesions of AIDS with CMVR have distinct features.
Objective To analyze the characteristics of images of fundus fluorescein angiography (FFA) in patients with central serous chorioretinopathy (CSC).Methods We observed the positions of leakage spots of retinal pigment epithelium (RPE) and analyzed their characteristics in 598 CSC patients. The ages, visual acuty, detachment areas of sensory retina, and leakage patterns of RPE were recorded and the correlation of them were analyzed. Results The leakage concentrated in the posterior pole, which occurred more frequently in the superior quadrant, nosal quadrant, and near fovea regions than in the inferior quadrant,temporal quadrant, and far regions from fovea, respectively (chi;2=67.13,20.93, 212.715; Plt;0.01).We found that there were relations between the ages of patients and the patterns of the leakage. A total of 35 patients (54.7% out of 64) at the age of ge;50 had small leakage; 82 patients (53.2% out of 154) at the age of 35-39 had diffusing leakage. Smaller retinal detachment in patients with multiple serous leakage and bigger retinal detachment area in patients with single serous leakage were found; no statistical significance was found (F=1.925,Pgt;0.05). There was negative correlation between the visual acuity and the detachment area in CSC patients (t=-0.335,Plt;0.01), and there was no correlation between the visual acuity and the distances from the leakage spots to the fovea (t=-0.029, -0.145;Pgt;0.05).Conclusion The leakages occurs in various regions whose difference is significant; the patterns of leakage are related to the ages; there is no relation between the serous retinal detachment area and the number of the leakage.
Objective To investigate the relationship between the level of testosterone and estradiol in serum and central serous chorioretinopathy (CSC).Methods The clinical data of 200 patients with active phase CSC who diagnosed by clinical manifestation, examination of fundus and fluorescence fundus angiography (FFA), were retrospectively analyzed. Two hundreds healthy people were collected as a control group. The blood of ulnar vein was collected and the method of magnetic homogeneous enzyme immunoassay was used to detect the level of testosterone and estradiol in serum of two groups. The results were analyzed statistically by t test.Results The values of testosterone and estradiol of male were all higher in CSC group than that in control group，the differences were statistical significance（t=2.804，2.913；P=0.010，0.008）；it was also higher in female（t=2.078，2.807；P=0.049，0.010）. The value of testosterone/estradiol of male was higher than that of female in CSC group，the difference was statistical significance(t=2.231，P=0.046).Conclusions The level of testosterone and estradiol in serum of CSC group increased obviously, especially the value of testosterone/estradiol. The increase of estradiol and testosterone/estradiol may be an etiological factor of CSC.
Objective To investigate the feasibility of gene transfection into retinal pigment epithelial (RPE) cells and photoreceptors (PRs) in vivo electroporation. Methods A total of 147 Sprague-Dawley (SD) rats were divided into 5, 10, 15, 20, 25, 30 and 35 V group according to different voltage. The right eyes of rats underwent the injection of eukaryotic expressive plasmid of enhanced green fluorescent protein (EGFP) pEGFP-N1 into subretinal space as experimental eyes; the left eyes were injected with TE buffer as control eyes. Each group was divided into RPE and RP subgroups according to different transfection direction. There were same parameters of 99 ms pulse width, 0.5 s pulse interval and 5 consecutive pulses except different voltage in groups. With a negative charge in the electric field was transfected into RPE cell layer, reverse electrode set to be transfected into PR cell layer. Retina mounts were made on seven days after transfection and the fluorescence of EGFP was photographed by fluorescent microscope. The expression of EGFP mRNA and protein were detected by reverse transcription polymerase chain reaction technique (RT-PCR) and Western blot.Results On seven days after transfection, in RPE subgroups, there were no specific fluorescence expressions in RPE cell layer and retina mounts of control eyes, while there were fluorescence expressions in experimental eyes. Western blot showed that the grayscale ratio of EGFP protein and beta;actin protein bands rose with the increased voltage. RT-PCR showed that each group produced positive amplification bands, and the relative ratio of gray level of EGFP mRNA and GADPH mRNA amplified bands gradually increased with the increased voltage.Conclusion Electroporation is an effective method for gene delivery into RPE cells in vivo.
Objective To observe ocular manifestations of acquired immunodeficiency syndrome（AIDS）.Methods Fourtytwo AIDS patients（66 eyes）with ocular complaints received examinations of visual acuity, slit-lamp microscope, ophthalmoscope and fundus fluorescence angiography (FFA). The results were retrospectively analyzed. Results There are five types of ocular findings, including cytomegalovirus (CMV) retinitis (37 eyes, 56.0%), retinal microvasculopathy of human immunodeficiency virus (21 eyes, 32.0%), optic nerve diseases (three eyes, 4.5%), retinal neuroepithelial layer detachment (two eyes, 3.0%) and uveitis (three eyes, 4.5%).Conclusions The common ocular manifestations showed progressive necrotic retinitis, retinal hemorrhage and retinal vasculitis and attenuated，cotton-wool spots in AIDS patients.