PURPOSE:In search of the mechanism for photic retinal injury. METHODS:A visible light damage model was established in the primary cultured healthy,adult human RPE cells by using intense fluorescence light (2 400 Lx). RESULTS:Electron microscopy revealed swelling of the mitochondria and obscurity of nuclear membranous structure in the light damaged cells. The decrease or dissolution of organelle,vacuolization of cytoplasm and myelinic degeneration were found in some severely damaged cells. The level of intracellular SOD was decreased to 41% of that of the controls (P＜0.05). CONCLUSION:The structure of the RPE was damaged by the light radiation and the level of intracellular SOD was decreased. These suggested the light damage might be associated with the production of free radicals and the lipid perioxide reaction in membranous structure of cell. （Chin J Ocul Fundus Dis,1996,12: 174-175 ）
ObjectiveTo investigate the effectiveness of arthroscopic treatment of pigmented villonodular synovitis (PVNS) of the ankle. MethodsTwelve patients who were initially diagnosed as having PVNS of the ankle were treated between January 2005 and May 2012.There were 6 males and 6 females,aged 20-50 years (mean,35.4 years).Disease duration ranged from 6 months to 12 years (median,3.6 years).One case of recurrence was included.The preoperative American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score was 55.5±7.6.According to degree and range of the PVNS lesions,4 cases of local PVNS were treated with arthroscopic debridement,and 8 cases of diffuse PVNS were treated with arthroscopically assisted arthrotomy;and local radiotherapy was given in all patients after operation. ResultsPrimary healing of incision was obtained in all patients.The mean follow-up time was 2.8 years (range,1-6 years).At 12 months after operation,no obvious pain,swelling,and limited range of motion of the ankle were observed.The AOFAS score was increased to 84.3±3.4 at 12 months,and it was significantly higher than that at preoperation (P<0.05) and at 3 months after operation (82.8±3.8)(P<0.05).There was no recurrence during follow-up. ConclusionArthroscopic arthrotomy combined with postoperative radiotherapy are recommended for PVNS of the ankle according to the PVNS lesion degree and range.And arthroscopically assisted surgery has many advantages of less traumas and hemorrhage,fast recovery,and less complications.
Objective To investigate the histological and keratinous variation of prefabricated urethra in the capsule with micro-mucosa and gelatin sponge compound graft. Methods Five 8-week-old Guizhou miniature pigs (2 females and3 males) weighing 20-25 kg were used. Eight tissue expanders were bilaterally inserted into subcutaneous position on the dorsal thorax of each pig. Forty inserted expanders were randomized into two groups (n=20 per group). For the experimental group, the free buccal mucosa was cut into particles less than 1 mm in diameter, spread onto the gelatin sponge (3 cm × 2 cm) and then transplanted to the capsule; the area expansion ratio of autogenous micro-mucosa was 8 ∶ 1. For the control group, soft tissue expander without mucosa graft was implanted. The pressure in inserted expander was about 40 mm Hg (1 mm Hg=0.133 kPa). Inflation should be stopped when the injected sal ine volume reached 15 mL. The animals were killed 1 and 2 weeks and 1, 2, and 4 months after the implant to receive examination. Macroscope, histology, and immunohistochemistry changes were observed. Results All the animals survived to the end of the experiment and the wounds healed by first intention. There was no obvious degeneration of gelatin sponge, and some of the mucosa survived 1 week after implant. The gelatin sponge was partly absorbed, most of the mucosa survived 2 weeks after implant. Visual examination showed complete epithel ial ization of the entire cavity 1 month after implant. The experimental group at 2 and 4 months were similar to that of at 1 month in gross observations.The neo-mucosa was not found in the control group at different time points after implant. Histology examination revealed that compound implant was mainly infiltrated by inflammatory cells and the micro-mucosa survived well 1 week after implant in the experimental group. The stratified squamous epithel ium presented obvious polarity and the submucous neovascularization was abundant 2 weeks after implant. The compound implant achieved complete epithel ial ization 1 month after implant. The epithel ium degeneration occurred 2 months after implant. The stratified squamous epithel ium presented no abovious polarity 4 months after implant. No neo-mucosa was evident in control group at different time points. The experimental group was positive for the pan-cytokeratin staining at 1, 2 weeks, and 1, 2 months after implant, but negative at 4 months after implant The pan-cytokeratin staining was negative in the control group at different time points. Conclusion The buccal micromucosa and gelatin sponge compound graft can grow well on the expanded capsule 1 month after implant and the epithel ium degeneration is evident 2 months after implant. Environment of implanted mucosa has great influence on epithel ium mucosa.
Objective To investigate the protective effect of nerve growth factor (NGF) on apoptosis of cultured human fetal retinal pigment epithelium (HFRPE) cells induced by indomethacin (IN) in vitro.Methods Subcultured HFRPE cells were treated with different concentrations of IN to establish apoptotic model. The protective effect of NGF on apoptosis of cultured HFRPE cells were assessed using an acridine orange (AO) staining method and transmission electron microscopy (TEM).Results HFRPE cells exposed by 200-600 μmol/L IN for 24 hours elicited typical apoptosis morphological changes, including condensed chromation, nuclear fragmentation and reduction of nuclear size and cell volume. There was a statistically difference in HFRPE cells with apoptosis between 200 μmol/L IN+500 μg/L NGF and 200 μmol/LIN groups ( q=3.9204，P=0.0320); there was a significant difference in HFRPE cells with apoptosis in 400 μmol/L IN+500 μg/L NGF and 400 μmol/ L IN as well (q=9.7915，P=0.0001). Conclusion NGF has an protective effect on IN-induced HFRPE cells apoptosis. (Chin J Ocul Fundus Dis，2003，19：38-41)
Purpose To investigate retinoic acid (RA) induced apoptosis in retinal pigment epithelial (RPE) cells. Methods 10-5、10-6、10-7 mol/L were added to cultured PRE cells.Aridine orange fluorescence and TdT-mediated dUTP nick end labelling(TUNEL) techniques were used to observe apoptotic changes. Resultss 10-5、10-6、10-7 mol/L RA induced apoptosis in RPE cells.Cell shringkage,chromatin condensation and nuclear DNA fragmentation of RPE cells were observed by TUNEL technique.When 10-7、10-6、10-5mol/L RA treated RPE cells for 5 days,apoptotic index(AI)was 36.9%、4409% and 61.4% respectively,and 48.0%、59.9%、74.2% for 6 days.At the same concentration of RA,AI increased when time prolonged．At the same day,AI increased when the concentration of RA rose.There was significant difference in the results(Plt;0.05). Conclusion Our results showed that RA-induced apoptosis in RPE cells was detected with a good dose and time response. (Chin J Ocul Fundus Dis,1998,14:153-155)
【Abstract】 Objective To explore the effect of early scrotal dermatoplasty on spermatogenic functional rehabilitation of testis in juvenile pigs with third degree burn wound of the scrotum. Methods Thirty healthy male Guizhou miniature pigs (weighing 10-15 kg, 2-month-old) were divided into 3 groups: control group (group A, n=10), natural healing group (group B, n=10), and dermatoplasty group (group C, n=10). In group A, the pig was not given any treatment; after third degree burn model of the scrotum was prepared, wounds were not treated in group B and the burn skin was excised and whole hypogastric pachydermia was used for dermatoplasty in group C. At 3 months and 1 year after model preparation, bilateral testis were collected from 5 pigs, respectively. HE staining was performed to observe the effects of different repair method on the morphology of spermatogenic cells and immunohistochemical staining was used to detect Survivin protein expression. Results All pigs survived to the end of the experiment and the wound healed successfully. Histological observation showed that spermatogenic cells had normal shape at all stages and mature sperms were seen in lumens in group A; the thickness of seminiferous epithelium was thinner, having one layer or two layers of spermatogenic cells in group B; the spermatogenic cells in group C were slightly more than that in group B with some spermatids; and in groups B and C, the spermatogenic cells at 1 year were more than that at 3 months. Immunohistochemistry staining showed that the Survivin protein expression in groups B and C was less than in group A, and group B was less than group C, showing significant differences at 3 months and 1 year (P lt; 0.05), but no significant difference between 3 months and 1 year in the same group (P gt; 0.05). Conclusion Dermatoplasty has inhibitory effect on spermatogenic functional rehabilitation of testis. Dermatoplasty can decrease spermatogenic cells and reduce Survivin protein expression, but some spermatids still survive in seminiferous tubule.
Objective To investingate the ultrastructural changes of retinal pigment epithelium(RPE) and its permeability in spontaneously hypertensive rats(SHR)and explore the relation between these changes and hypertensive retinopathy.MethodsThe ultrastructure of RPE cells in the SHR aged five,six,seven months wasobserved with transmission electronmicroscope and compared to its normotensive control strain(WKY) with the same age.Then,lanthanum tracer procedures were carried out to investigate pathological changes of the blood-retinal barrier.Results (1)In SHR the main pathological changes involved swelling of mitochondria,enlargement of endoplasmic reticula,decrease of RPE cell infolding,and sparseness of microvilli.These degenerations were more serious in older rats with higher blood pressure.(2)The breakdown of outer blood-retinal barrier with permeation of lanthanum tracers were evident in SHR aged six or seven month,however,in WKY and five-month SHR the traces were prevented from passing by tight junctions.ConclusionThe degeneration of RPE owing to ischemia and anoxia arises in early periosd of hypertensive retinopathy.The pathological changes of ultrastructure and permeability might interact with the damage of visual cells and play a main role in the hypertensive retinopathy.
Purpose To investigate the effects of human vitreous fluid on proliferation of cultured human retinal pigment epithelial (RPE) cells and vascular endothelial cell lines(VEC304). Methods Human RPE cells and VEC304 were cultured and treated in different human vitreous-conditioned medium (VCM) with or without serum, vitreous volume concentrations of VCM were 1∶8, 1∶4 and 1∶2. Cells proliferation was assayed by tetrazolium (MTT) colorimetry at the 2nd, 4th and 6th day respectively. Results In the presence of serum, 1∶4, 1∶2 VCM had a significantly stimulative effect on RPE cells proliferation compared with control group at the 2nd, 4th, and 6th day retrospectively (P＜0.01), but exerted a bly inhibitory effect on VEC304 proliferation compared with control group at the 2nd, 4th, and 6th day retrospectively (Plt;0.05). In the absence of serum, only 1∶2 VCM had a stimulative effect on RPE cells growth compared with control group at the 2nd day (P＜0.05) and obviously at the 4th and 6th day respectively (P＜0.01). Conclusion Human vitreous fluid influences human RPE cells and VEC304 growth in vitro. This result suggests that vitreous may play different role in proliferative vitreoretinopathy and intraocular neovascular disease. (Chin J Ocul Fundus Dis, 2002, 18: 140-142)
Porpose To investigate the optimal concentration of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) on DNA synthesis and their synergism indensity arrested human retinal pigment epithelial (RPE) cells. Methods Growth factor effects in cultured human RPE of the 6th generation were assessed by [3 H]-thymidine incorporation and radioautography. Results EGF and bFGF were potent stimulators when used alone,and their optimal concentrations were 10ng/ml in DMEM and 1ng/ml in 2% serum DMEM.When used in combination (10ng/ml EGF and 10ng/ml bFGF),they caused a significant enhancement of [3 H]-thymidine incorporation about 2.96 times. Conclusion EGF and bFGF were potent stimulators in RPE cells,and demonstrated synergism in their action. (Chin J Ocul Fundus Dis,1998,14:98-100)