Objective To establish a hemorrhagic retinal detachment (HRD) model for the study of the damage and treatment of HRD.Methods Fourteen rabbits (28 eyes) were divided into the HRD (12 eyes) and control (16 eyes) group randomly. Autologous anticoagulated blood (0.2 ml) was transvitreally injected into the rab bits′subretinal space with a special glass micropipette in HRD group （12 eyes ）; while 0.2 ml saline with or without heparin sodium （2.5 U/ml） was respectively injected into subretinal space respectively of the rabbits in heparin saline control group (6 eyes) and saline control group (3 eyes); furthermore, another 2 control groups, i.e.,pseudo injection group (3 eyes, single retinal puncturing without subretinal injection) and normal group (4 eyes of 2 normal rabbits) were also set. The conditions of the occurrence and representation of the retinal detachment (RD) were observed and analysed by means of ophthalmoscopy, optical coherence tomography (OCT) and ultrasound A and (or) B scan examinations in the subsequent 28 days after the operation.Results After the operation, HRD occurred in all eyes of the rabbits in HRD group. The area of HRD extended from 10 to 12 disc diameter(DD). The obvious elevation of RD maintained to 14 days, and the residual subretinal hemorrhage was still observed till 28 days. The obvious RD of the rabbits in hepar in saline and saline control group was only kept for no more than 12 hours. The retinal puncture hole in pseudo injection group disappeared 2 days after the operation, and there was no change in retina of rabbits in normal control group.Conclusion It is convenient, practical and effective to establish a HRD model by means of transvitreal subretinal injection of autologous anticoagulated blood. (Chin J Ocul Fundus Dis，2003，19：175-178)
Objective To determine the effect of methimazole (MMI) on retinal vascular development in neonatal rats, and to investigate the relationship between the concentration of insulin-like growth factor-I (IGF-I) in serum and the development of normal blood vessels and between the concentration of IGF-I and the formation of abnormal blood vessels. Methods There were 75 neonatal SpragueDawley rats in experimental group whose mothers were raised with water with 0.1% MMI at the first day of parturition. Another 50 neonatal rats were in the control group whose mothers were raised with normal water. The rats in the two groups were sub-divided into 4day and 10day subgroup, respectively. The retinal flatmount of the right eyes were stained with adenosine diphosphatase (ADPase); with the paraffin section of the left eyes, the number of nucleolus breaking through retinal inner limiting membrane was counted and the retinal blood vessels were evaluated. Serum IGF-I levels were detected by radioimmunoassay, and the weight of the neonatal rats in each group were observed and recorded. Results The incidence of retinal neovascularization in 10 day MMI group was 27%, and 0% in 4-day MMI group and control group. The serum IGF-I level in 4-day and 10-day MMI group (73.07 ng/ml, 175.13 ng/ml) was obviously lower than which in the 4-day and 10-day control group (168.73 ng/ml,306.38 ng/ml) （P=0.00）. Obvious slow growth of the neonatal rats was found in MMI group compared with which in the control group. Conculsions MMI may inhibit the normal growth of retinal blood vessels and lead neovascularization, which may relate to the initial decrease of the serum IGF-I level. (Chin J Ocul Fundus Dis, 2007, 23: 198-201)
Objctive To explore the relationship between the expression of Fas/FasL and the apoptosis occurs in retinal ischemia/reperfusion injury of rats , as well as the therapeutic effects of bFGF on the ischemic retina.Methods Th emodels of retinal ischemia/reperfusion injury was made by transient elevating introcular pressure. A total of 28 rats were divided into normal and operation group.The latter were subdivided into 1 hour, 6, 12, 24, 48 and 72 hours after reperfusion group, in which the left eyes of the rats were in the ischemia/reper fusion groups and the right ones were in the treatment groups (bFGF intracameral injection). Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) method, and the expression of Fas and Fas ligand was studied by strept avidin-biotin complex （SABC）immunohistochemistry. Results No positive cells were observed in the normal rats′retinae, but there was a significant number of TUNEL positive cells in 6-24 hours after transient ischemia followed by a decrease at the 48th hour. The number of TUNEL positive cells reached a maximum at the 24th hour after ischemia. The expression of Fas gradually increased as early as when it was at the 6th hour, reached a peak at the 24th hour, and then decreased at the 48th hour. Similarly, the expression of Fas ligand was at peak in 24-48 hours in GCL and INL of retina. Conclusions Retinal ischemia-reperfusion after transient elevated IOP induced apoptosis of cells in the retina. Fas/FasL may play an important role in the early events of the apoptotic pathways. bFGF can rescue RGCs from retinal ischemia/reperfusion injury through downregulation of the expression of Fas/FasL and may represent an important mechanism for therapeutic neuroprotection. (Chin J Ocul Fundus Dis，2003，19：160-163)
ObjectiveTo investigate the neuroprotective effect of Benztropine on retinal ganglion cells (RGCs) death and optic nerve injury in rats model of non-arteritis anterior ischemic optic neuropathy (rNAION).MethodsA total of 25 Sprague-Dawley rats were randomly divided into Benztropine treatment group (n=13) and PBS control group (n=12). The right eye was set as the experimental eye. rNAION model was established by using rose Bengal combined with laser photodynamic method. The rats in the Benztropine treatment group were received intraperitoneal injection with Benztropine 10 mg/kg (0.2 ml) daily for 3 weeks, while the rats in the PBS control group were received intraperitoneal injection with an equal volume of PBS. At 1, 3 and 7 days after modeling, the retinal and optic disc conditions of the rats were observed by direct ophthalmoscopy. Retrograde labeling, fluorescence microscopy and transmission electron microscopy were used to observe the survival of RGCs and the damage of the optic nerve myelin and axon at 4 weeks after modeling. The RGCs density and survival rate of the two groups were compared by One-Way Anova.ResultsAt 1 and 3 days after modeling, the optic disc edema was observed in the rats of rNAION model group. At 7 days after modeling, the optic disc edema decreased and the boundary was blurred compared with 3 days after modeling.After 4 weeks, the RGCs density in the PBS group was 308±194/mm2 and the survival rate was 13.7%. The density of RGCs in the Benztropine group was 1173+868/mm2 and the survival rate was 47.6%. The differences of RGCs density and survival rate were significant between the two groups (F=7.552, 8.184; P=0.015, 0.012). Myelin disintegration, axon degeneration, onion-like body and gliosis were observed in the optic nerve sections of rNIAON in the PBS group, while the damage of axon and myelin structure in the Benztropine group was significantly less than that in the PBS group.ConclusionsBenztropine group showed higher RGC survival rate, less damage of axon and myelin structure on rNAION model. This study explored the potential neuroprotective effect of Benztropine.
Objective To investigate the response of retinal ganglion cells （RGC）in detached and reattached retina in adult rats, and the effect of IL-1beta antibody and IL-1Ra on the loss of RGC. Methods A total of 73 Sprague-Dawley (SD) rats were subretinally injected with healon GV（1.4% hyaluronate）and retrograde labeled with fluorogold (FG), and 10 ng IL-1 Ra and 500 ng IL-1beta antibody were injected into the subretinal space combined with healon GV. The retinal flakes were observed under the fluoroscope and the number of RGC was counted 2 hours, 1, 3, 5, 7, 10, 20, 50, and 90 days after deta chment; 10 days after detachment and 30 days after reattachement; 90 days after detachment and 20 days after reattachement, and 1 and 10 days after injection with IL-1beta antibody and IL-1Ra,respectively. And the control group was only developed an intraocular injection of the same valume of healon GV. Result Two hours after detachment, the RGC loss was found, reached the peak at first day, and decreased gradually. RGC loss was also found in the non-detached area. The reattachment 10 days after detachment (early reattachment) stopped the loss of RGC, and the reattachment 90 days after detachment (late reattachment) promoted the loss, which rested on a certain level. Subretinal space injection of IL-1Ra and IL-1beta antibody decreased the loss of RGCs in the detached retina. Conclusion The RGCs loss were found both in the detached and attached retina. Early reattachment may stop the loss of RGC, and late reattachment may promote the loss. Both IL-1beta antibody and IL-1Ra have neuro protective effect on RGC. (Chin J Ocul Fundus Dis，2004，20：233-236)
Objective To establish an allogenic intraocular melanoma model and observe its pathological features.Methods Thirty-six kunming mice were devided randomly into 3 groups with 12 ones in each, and allogeneic melanoma cells B16F10(C57BL16) were inoculated into the anterior chamber (AC), vitreous cavity (VC) of right eyes and under the skin (subcutaneous, SC) of the back of right feet of each grup respectively. The incidence of tumor occurance, time of breaking through the eyeball and other general pathologic features of the tumor were observed by slip-lamp biomicroscopy and operating microscopy for continuous 32 days, and the results were statistically analyzed. Pathological examination was given for tumors at last.Results The incidence of tumor occurance in both AC (12/1 2 eyes) and VC group (11/11 eyes) was higher than that in SC group (2/12 feet)（χ2=17.143, P＝0 .000；χ2=16.218, P=0.000）. The time of eyeball diabrosis was 11-13 days in AC group and 13-32 days in VC group, and there was significant difference between these two groups (Log Rank=18.22, P=0.000). The intraocular melanomas could grow progressively, but reduced and fell off when they broke through eyeball and grew in or bit for a period. The average diameter of the tumor after 32 days after inoculation was (2.27±1.97) mm in AC group，(3.82±1.85) mm in VC group and (0.94±2.27) mm in SC group. There was significant difference between VC and SC group (t=3.322，P=0.003). In pathohistological examination, tumor tissue necrosis could be observed at the center of the subcutaneous melanomas but not in intraocular melanomas.Conclusions Allogeneic intraocular melanoma model is successfully established which is convenient, repeatable, and helpful to studying the mechanism of genesis and development of this tumor. (Chin J Ocul Fundus Dis，2003，19：333-404)
Objective To evaluate the effect of vascular endothelial cell growth factor (VEGF) antisense oligodeoxynucleotides (ASODNs) on the expression of VEGF in rats with oxygen-induced retinopathy. Methods Thirty newborn Sprague-Dawley (SD) rats were randomly divided into 3 groups:normal control group, disposal group and non-disposed group, The animal models with oxygen-induced proliferative retinopathy were established by raising the rats in hyperoxic environment. Retrobulbar injection was performed with VEGF ASODNs or normal saline on the rats in 3 groups respectively. The intraocular tissues (all the tissues except the cornea, sclera, and lens) and serum were collected, and the expressions of VEGF were determined by using competitive enzyme immunoassay.Results The expressions of VEGF in intraocular tissues of rats in disposal group were significantly lower than those in non-disposed group (P＜0.05), and there was no statistical difference between the disposal and normal control group (P＞0.05). There was no significant difference of the expressions of VEGF in serum of rats between the disposal and non-disposed group (P＞0.05), which were both lower than those in the normal control group (P＜0.05). Conclusion VEGF ASODNs could significantly inhibit the expression of VEGF in intraocular tissues. (Chin J Ocul Fundus Dis，2003，19：172-174)
Objective To observe the inhibitory effect of Bevacizumab on retinal neovascularization in oxygen-induced retinopathy in the mouse. Methods 90 one-week-old C57B L/6J mice were divided into four groups at random. 15 mice in the 1st group as normal control group, 15 mice in the 2nd group as oxygen control group, 30 mice in the 3rd group as high-dose Bevacizumab treatment group, 30 mice in the 4th group as low-dose Bevacizumab treatment group. The 2nd, 3rd and 4th groups were exposed to 75% oxygen for 5 days and then to room air. At the 12th day, One eye of each mouse of two control groups were received an intravitreal injection with Be vacizumab at 2 mu;l、1 mu;l respectively, and the same volume of BSS was injected into the other eye of the mice. The adenosine diphosphatase (ADPase) histochemical technique was used for retinal flat mount to assess the oxygen-induced change s of retinal vessels. The number of the endothelium cell nuclei of proliferative neovascularization was quantified by retinal microtome chromoscopy. Real-time PCR analysis was performed to examine the expression of VEGF mRNA. Results Comparing with oxygen control group, regular distributions, reduced density of retina l vascular and reduced endothelium cell nuclei which extending retinal membrane were observed in the treatment groups(P＜0.001). But the differences between two treatment groups are not statistically significant (P＞0.05). The expression of VEGF mRNA was not significantly different in oxygen control group whatever it whether accepted Bevacizumab treatment or high or low dose (P＞0.05). Conclusion Intravitreal injection with Bevacizumab can effectively inhibits the retinal neova scularization in oxygen-induced retinopathy in the mouse. Intravitreal injection with Bevacizumab might become to the new method to treat retinopathy of premature. （Chin J Ocul Fundus Dis，2008，24：184-188）
ObjectiveTo evaluate the protective effect of estrogen on survival of retinal ganglion cells (RGCs) after transient retinal ischemia-reperfusion (RIR) in rats.MethodsRIR was induced in 60 ovariectomized adult rats (OVX) by increasing intraocular pressure via an intracameral catheter. All of the rats were divided into two groups randomly: in experimental group, the rats underwent a subcutaneous injection with 17β-estrodiol（100 μg/kg） 2 hours before retinal ischemia; and in the control group, saline water was injected correspondingly. The number of RGCs and the thickness of the inner retinal layers were mesured by HE staining method before and 12, 24, 48, and 72 hours after reperfusion. TdT-mediated biotin-dUTP nick end labelling (TUNEL) staining technique was used to examine the apoptosis of RGCs.ResultsTwenty-four and 48 hours after reperfusion, the number of apoptotic cells in experimental group was obvious lower than that in the control group(Plt;0.05), and the number of RGCs in experimental group was higher than that in the control group(Plt;0.05).ConclusionEstrogen can protect retinal neurons from transient RIR in ovariectomized rats.(Chin J Ocul Fundus Dis, 2005,21:177-179)
Objective To investigate the expression of T cell receptor (TCR) Vβ8.3 gene on CD4+ T lymphocytes in the rats with experimental autoimmune uveoretinitis (EAU). Methods Eighteen Lewis rats were divided into EAU, complete Freund′s adjuvant, and the control group. Inter photoreceptor retinoid-binding protein (IRBP) R16 peptide was synthesized using Fmoc procedure for induction of EAU. Magnetic absorption cell sorting (MACS) me thod was used to isolate the CD4+T lymphocytes from the spleen of the rats. Flow cytometry was used to monitor the efficiency of isolation. The expression of TCR Vβ8.3 gene segment on CD4+T lymphocytes was determined by fluorescent quantitative polymerase chain reaction. Results EAU was successfully induced in the Lewis rats immunized with IRBP R16 peptide. The proportion of CD4+T lymphocytes isolated by means of MACS was statistically higher than that before isolation (P＜0.001). The expression of TCR Vβ8.3 gene segment on CD4+ T lymphocytes in EAU rats was significantly higher than that in the control (P＜0.05). Conclusions There is a predominant usage of antigen-specific TCR Vβ 8.3 gene in EAU rats induced by IR BP R16 peptide, which may serve as a target for immunotherapy of EAU. (Chin J Ocul Fundus Dis，2004，20：165-167)