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 study the effect of splenectomy on the anti-tumor immunity in rats with induced hepatocellular carcinoma (HCC). Methods At the second and fourth month of the induced HCC, the NK cell activity, TNF-α level and total lymphcyte in blood were measured in the group of splenectomy and the control group. Results There were no different in the total lymphcyte and TNF-α in the blood in two groups, but there were significant difference in the NK cell activity between the group of splenectomy and the control group (Ｐ＜0.05). Conclusion There are some change in the anti-tumor immunity after splenectomy in rats, in which NK cell activity is at low level continuously. TNF-α isn′t affected after the second month after splenectomy.
ObjectiveTo observe the expression of interleukin (IL)-17, IL-4 and interferon γ (IFN-γ) in experimental autoimmune uveoretinitis (EAU). MethodsC57BL/6 mice were immunized with interphotoreceptor retinoid-binding protein 1-20 to induce EAU. The inflammatory reaction before and on 7, 14, 21, 28 days after immunization were observed. The level of IL-17, IL-4 and IFN-γ in the serum were measured by enzyme-linked immune sorbent assay (ELISA). mRNA and protein expression of spleen and retina were analysed using quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot at the same time, respectively. ResultsThe most serious inflammatory reaction occurred at the 14th day after immunization. The highest level of IFN-γ in serum, highest mRNA and protein expression of IFN-γ in spleen and retina of mice occurred at day 7 after being immunized. The highest level of IL-17, IL-4 in serum, highest mRNA and protein expression of IL-17, IL-4 in spleen and retina of mice occurred at day 14 after being immunized. The increase degree of IL-17 was more than IFN-γ and IL-4. At 7, 14 and 21 days after immunization, compared with the pre-immunization, the level of IL-17, IL-4, IFN-γ in serum of mice were significantly increased (F=1 817.346, 268.600, 164.621; P < 0.05). There was no difference in the levels of IL-17, IL-4, IFN-γin serum of mice between pre-and 28 days after immunization (P > 0.05). At 7, 14 and 21 days after immunization, compared with the pre-immunization, the protein expression of IL-17, IL-4, IFN-γ in spleen (F=312.67, 114.250, 216.220) and retina (F=271.504, 85.370, 80.722) of mice were significantly increased (P < 0.05). There was no difference in protein expression of IL-17, IL-4, IFN-γ in spleen and retina of mice between pre-and 28 days after immunization (P > 0.05). ConclusionsThere were IL-17, IL-4 and IFN-γ expression in EAU. IL-17, IL-4 and IFN-γ play a key role in the occurrence and development of the EAU.
Taking Wistar rats and pigs as models, the basic fibroblast growth factor (bFGF) was studied on wound healing. Five rats and five pigs were used in the experiment. Each rat had 2 roundshapedwound (1.8cm in diameter) and each pig had 4 wounds of the same size as that ofthe rat. One wound of each rat and 2 wound of each pig were dressed with bFGF saline (60U/cm2). While the other wounds of the rats and pigs were dressed with normal saline as control. The wounds treated with bFGF were completely covered with granulation tissue on the 7th day after injury, and in 14 days the wounds were almost completely covered by epithelium. The bFGF would enhance the growth of theepithelial cells, reepithelization of the wound and the tensile strength of thetissue. It was concluded that the bFGF could promote skin regeneration, whichmight be the direct action of bFGF on the bFGF receptors in the basic cells of skin.
Objective:To observe the effect of beta;estradiol on gluta mate concentration in rabbitsprime; retinae injured by ischemic reperfusion. Methods:Twenty r abbits ware randomly divided into two groups, the control group and the treatmen t group, with 10 rabbits in each group. Before examined by binocular flash elect roretinography (FERG), retinal ischemic reperfusion (RIR) model was induced in t h e right eyes of all the rabbits by increasing intraocular pressure to 120 mm Hg for 60 minutes; the left eyes were as the control eyes. The rabbits were hypoder mically injected with beta;estradiol (0.1 mg/kg) in treatment group and with phys i ological saline in the control group 2 hours before ischemia. The results of FER G of the right eyes in both of the 2 groups 0, 4, 8, and 24 hours after reperfus ion were record respectively and were compared with the results of FERG before r eperfusion. The retina tissue was collected after the last time of FERG. The con c entration of glutamate was detected by Hitachi L8800 amino acid analyzer. Results:In the right eyes in both of the 2 groups, the result of F ERG showed a beeli ne just after reperfusion. There was no significant difference of awave amplit u de between the 2 groups (t=1.357, 0.798, 0.835; Pgt;0.05); the b wave amplitudes i n experimental group were much higher than those in the control group (t=4.447, 2.188, 3.106; Plt;0.01). The concentration of glutamate in retina was (0.265plusmn;0.014) g/L in the right eyes and (0.207plusmn;0.013) g/L in the left eyes in the control group, and (0.231plusmn;0.007) g/L in the right eyes and (0.203plusmn;0 .014) g/L in the le ft eyes in the treatment group; the difference between the 2 groups was signific ant (F=50.807, P=0.000). There was statistical difference between righ t and left eyes both in the 2 groups and the significant difference of the right eyes betw een the two groups was also found (P=0.000); there was no statistical diffe rence of the left eyes between the 2 groups (P＝0.505). Conclusion:beta;-estradiol may prevent the increase of the concentration of glutamate in retina induced by RIR to protect retinal tissue.
Natural nonorganic bone(NNB) was obtained after the fresh bone of pig was heated to 100℃. The NNB was white and in a shape as its original bone.The tensile strength of the compact bone was 200kg/cm3 and that of the cancellous bone was 25kg/cm3. The ratio of calcium to phosphorus was 10∶6. The main componentwas hydroxyapatite. The material was composed of trabeculae and intertrabecular spaces. Three experiments were performed. Experiment 1: 18 pieces of NNB in a size of 0.5×0.5×0.5cm3 were implanted intothe back muscle of 18 rabbits. At 4, 8 and 12 weeks after the operation, 6 specimens were obtained seperately and were stained by HE, and then examined under microscope. The result showed that the mesenchymal cells had no regeneration and differentiation, and the NNB and the surrounding tissues had no evidence of formation of new bone or chondrosynthesis. This NNB did not produce rejection reaction between tissues but the new blood vessels could easily grow into the space of the NNB. The fibrils had intimate contact with NNB. Experiment 2: The NNB and hydroxyapatite(HA) were mixed to make a cylindroid body with 2mm in diameter and 4mm in length and was implanted in the bilateral tibias of 40 rabbits, respectively. The roentgenography, fluroscent microscopic examination and histological observation were carried out at 2, 3, 4 and 8 weeks after the implantation. Experiment 3: In 50 rabbits, a defect of 2.5cm was made on both radius, and in one sideNNB was implanted and the other side was served as the control. Another 50 rabbitsHA was implanted in the defect in one side and the other side was served as the control. The results showed that in the NNB group at the 16th week, the bone united in 16 of the 30 cases, while in the HA group, in the 30 cases,only 2 had the bone united, while those of the controls no union had occurred.It was suggested that NNB had more formation of new bone than HA did.
Objective To investigate the effects of lights with different wavelength on the retina of rd12 and C57BL/6J mice. Methods Thirty two rd12 mice and C57BL/6J mice were randomly divided into the control group, white light group, midwavelength light (505 nm) group and shortwavelength light (405 nm) group, with eight mice in each group. Besides the control group, other groups were exposed to cycle illuminations ［12 hours dark, 12 hours (800plusmn;130) Lux］ for seven days to establish the model of retinal light damage. Electroretinogram (ERG) responses of all mice were recorded at the day before illumination and 1st, 4th and 7th days after illumination. The eyes were enucleated at 7th days after illumination to assess levels of reactive oxygen species (ROS), expression of peroxiredoxin 6 (PRDX6), and activity of caspase-3. Results ERG amplitudes of all groups declined gradually in C57BL/6J mice, and the most significant effects was found in the short-wavelength light group. The amplitudes of photopic b-wave were significantly different at 1st, 4th and 7th days (F=4.412, 5.082, 9.980；P＜0.01). The amplitudes of cone b-wave of the four groups decreased to (85plusmn;10) %, (70plusmn;19) %, (57plusmn;22) % and (46plusmn;19) % at 7th days, respectively, and were significantly different between white light group and short-wavelength light group(t=3.19,P＜0.01). The levels of ROS were significantly different in rd12 mice (F=16.08,P＜0.01), and elevated obviously in shortwavelength light group. The expressions of PRDX6 of retina were significantly different in rd12 mice (F=7.214,P＜0.05), and were decreased obviously in short-wavelength light group. The caspase-3 relative activity was significantly different in rd12 retina (F=7.530,P＜0.05); but there was no significant difference in C57BL/6J mice (F=3.625, 1.993, 1.133; P＞0.05).The caspase-3 relative activity were significant different between rd12 mice and C57BL/6J mice in short wavelength light group (t=5.474,P＜0.05). Conclusions Short-wavelength light can induce retinal damage of mouse retina, especially in rd12 mouse. The retinal light damage possibly relates to the oxidative damage.
Objective To investigate the effect of the traditional Chinese medicine “Tetrandrine”(TET) and its significance on epidermal growth factor(EGF) and its receptor(EGFR) in the lung of nitrofen-induced congenital diaphragmatic hernia(CDH) rat model. Methods Twenty female rats were given maternal administration of a single oral dose (115 mg/rat) of nitrofen to induce CDH at 9.5 days after pregnancy and were dividedinto normal solution group(NS, n=5), dexamethasone group (Dex, n=5),tetrandrine group (TET, n=5) and Dex+TET group(n=5) at 18.5 days; 4 rats were given edible oil as controls. All fetuses were delivered by cesarean section at 21.5 days. Lung histologic evaluations and EGF, EGFR immunohistochemical staining and image analysis were performed. Results CDH was observed in 64 of the 137 rat fetuses (46.7%) in the experimental groups; no CHD was observed in 36 rat fetuses of control group. The lungs of CDH fetuses showed marked hypoplasia in NS group, in contrast to improved mesenchymal differentiationin that of Dex, TET, Dex+TET groups. The expression of EGF was weaker and weaker and that of EGFR was ber and ber as following order: NS, TET, DEX, T+D and control groups; showing significant differences between them (Plt;0.05). Conclusion Prenatal TET administration shows marked improvement in pulmonary hypoplasia through preregulating crest-time of EGF expression and upregulating EGFR expression in the lungs of nitrofen-induced CDH rat model. A combination of TET and Dex would generate evident synergistic effect.
Objective To establish a rat model of blood ocular barrier breakdown induced by anterior segment intraocular analogic surgery. MethodsOne hundred and fifty healthy adult male rats were randomly divided into control group and model group, 75 rats in each group. The rats were anesthetized with 1 ml/kg ketamine hydrochloride/xylazine hydrochloride solution. Three way pipes were attached to a phosphate buffer infusion bag and two intravenous catheters. One catheter was inserted 30° obliquely through the transparent cornea anterior to the limbus into the rat's anterior chamber. Then the needle was withdrawn and the sheath was indwelling. Another catheter was connected with a manometer. Intraocular pressure was varied from 0 to 12 mm Hg (1 mm Hg=0.133 kPa) 60 times, 30 times per min. The catheter was removed. The eyes were treated with ofloxacin ophthalmic solution after surgery. The 1st, 2nd, 3rd, 5th and 7th day after surgery, the integrity of the blood ocular barrier was assessed by immunohistochemical staining for albumin and quantitative measurement using Evan′s blue as a tracer. ResultsAlbumin immunohistochemical staining of the control group was confined to the iris and retinal blood vessels. The choroid was stained at each time point after surgery. Albumin immunohistochemical staining of the model group was abundant around the iris and the retinal vasculature on the 1st day after surgery. The albumin diffused throughout the iris and the retina on the 2nd and the 3rd day after surgery. The albumin reached the retinal vessels on the 5th and 7th day after surgery. The aqueous humor Evans blue leakages of the model group were higher than those of the control group on the 1st, 2nd, 3rd and 5th day after surgery. The differences were statistically significant (t=25.781, 37.433, 25.150, 19.171； P<0.01). The Evans blue leakage of the model group was close to that of the control group on the 7th day after surgery. The difference was no statistical significant（t=1.303, P=0.209）. The retinal Evans blue leakages of the model group were higher than those of the control group on the 1st, the 2nd and the 3rd day after surgery. The differences were statistically significant (t=11.997, 14.622, 23.014; P<0.01). The Evans blue leakage of the model group was close to those of the control group on the 5th and 7th day after surgery. The differences were not statistically significant（t=2.027, 0.756; P=0.058, 0.459）. Conclusion This study establishes a rat model of blood ocular barrier breakdown induced by imitating the injury to the anterior segment during intraocular surgery.