Purpose:To investigate the dynamics of heat shock protein 72(HSP72) expression in retinal ganglion cells(RGCs) in rat model of acute glaucoma treated with heat stress or intraperitoneal injection of zinc sulfate.Metho...Purpose:To investigate the dynamics of heat shock protein 72(HSP72) expression in retinal ganglion cells(RGCs) in rat model of acute glaucoma treated with heat stress or intraperitoneal injection of zinc sulfate.Methods:Twenty-seven male Wistar rats were used to make acute glaucoma models. Five others served as normal control. Acute glaucoma models were made by intracameral irrigation in the right eyes with balanced salt saline (BSS) at 102 mmHg for 2 hours. Nine model rats were killed at different intervals after intracameral irrigation without treatment,which served as damage control. Ten were treated with heat stress 40℃~42℃, and 8 were used for zinc sulfate administration 2 days posterior to intracameral irrigation. Treated model rats were sacrificed at designed intervals after treatment. Right eyes were enucleated immediately, and the retinas were dissected for Western blot.Results:No HSP72 was found in RGCs of normal Wistar rats. In damage control group, slight HSP72 was detected during 6~36 hours posterior to intracameral irrigation. HSP72 was detected significantly expressed in RGCs of both heat shock group and zinc sulfate group. But the dynamics of HSP72 production were quite different in these two treated groups. In heat shock group, HSP72 appeared at the sixth hour after treatment, and increased gradually until its peak production emerged at the 48th hour. HSP72 vanished 8 days later after treatment. In zinc sulfate group, HSP72 expression began 24 hours later after zinc administration, and reached its highest level at the 72th hour posterior to treatment. HSP72 expression then decreased slowly, and disappeared 21 days later after treatment. Conclusion:HSP72 can be induced in RGCs of rat acute glaucoma models with heat stress or zinc sulfate adddministration. But the dynamics of the HSP72 induction in those two groups were quite different. Eye Science 2004;20:30-33.展开更多
Purpose: To investigate the effect of high dose methylprednisolone (MP) on retinal ganglion cells (RGC) apoptosis.Methods: One hundred and twenty six Wistar rats were divided into normal control, crush control and MP ...Purpose: To investigate the effect of high dose methylprednisolone (MP) on retinal ganglion cells (RGC) apoptosis.Methods: One hundred and twenty six Wistar rats were divided into normal control, crush control and MP treatment groups. High dose MP and 0.9% sodium chloride were injected intravenously at 1 h after injury in the treatment group and crush control group, respectively. Apoptotic cells, Bcl-2 and Bax positive cells were located and calculated by terminal-deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling (TUNEL), immunohistochemical method and retinal whole-mount combined with section technique at day 4, 7, 14 after crush, respectively.Results: The TUNEL-labeling cells, Bcl-2 and Bax positive cells were detected in the ganglion cell layer of retina. Few apoptotic cells, a few of Bcl-2 and Bax positive cells were observed in normal control groups. The numbers of positive cells increased gradually after crush. The number of apoptotic cells and Bax positive cells in treatment groups was less than that in crush group significantly (P < 0.05), while the number of Bcl-2 positive cells in treatment group was more than that in crush group at day 4, 7 and 14 after crush significantly (P < 0.05) Conclusions: These results suggest that high dose MP can inhibit the apoptosis of RGC after optic nerve crush probably through up-regulation of Bcl-2 expression and down-regulation of Bax expression.展开更多
Purpose: To investigate the effects of culture medium of human amniotic membrane (AM) on corneal neovascularization (CNV) induced by basic fibroblast growth factor (bFGF) in mice.Methods: Culture medium of amniotic me...Purpose: To investigate the effects of culture medium of human amniotic membrane (AM) on corneal neovascularization (CNV) induced by basic fibroblast growth factor (bFGF) in mice.Methods: Culture medium of amniotic membrane was prepared by cultivating AM (with epithelium side up) in EGM basic medium for 3 days, and was collected separately to three groups, e.g. Control (EGM only), AM with epithelium (AM) and AM without epithelium (De-AM). Corneal neovascularization was induced in mice by using micropocket assay with Hydron polymer pellets containing 100 ng bFGF. Migration and proliferation of human umbilical cord vein endothelial cells (HUVEC) were performed in Boyden chambers and by using the CyQUANT fluorescence binding assay respectively.The levels of tissue inhibitors of metalloproteinase 1 and 2 (TIMP1, TIMP2) in culture medium were determined by ELISA assay.Results: CNV induced by bFGF was significantly suppressed by culture medium of amniotic membrane. When the medium was applied as an eyedrop 4 times a day for 7 days,the area of CNV was (2.48±0.76) mm2,(0.64±0.52) mm2 and (1.96±0.65) mm2 incontrol, AM and De-AM group respectively. The migration and proliferation of HUVEC were strongly inhibited by culture medium of AM with epithelium, while the De-AM had no effect on the migration of HUVEC cells. The high level of TIMP2 was found in AM group, but not in De-AM group, while there was no difference in the amount of TIMP1 in medium among three groups.Conclusion: Culture medium of amniotic membrane significantly suppresses the corneal nevovascularization induced by bFGF. The mechanism of which at least in part is that high level of TIMP2 protein secreted or released into the culture medium of AM and inhibition of migration and growth of vascular endothelial cells.展开更多
Purpose:To determine whether cultured human trabecular meshwork cells express CD44 and to discuss their possible relationship with primary open angle glaucoma.Methods:Human trabecular meshwork cells were cultured in D...Purpose:To determine whether cultured human trabecular meshwork cells express CD44 and to discuss their possible relationship with primary open angle glaucoma.Methods:Human trabecular meshwork cells were cultured in DMEM/F12 media. Total RNAs from the cells were extracted with Trizol reagent. Messenger RNA expression of CD44 in human trabecular meshwork cells was examined by using reverse transcriptase-polymerase chain reaction ( RT-PCR ) analysis. Expression of CD44 was confirmed by Western-blotting and immunofluorescent microscopy. Effect of CD44-specific antisense oligonucleotide on adhesion of trabecular meshwork cells to hyaluronate was determined by MTT assay. Results:A single RT-PCR product whose size was 471bp was obtained. A band about 80kD was stained by Western-blot. Immunofluorescent examination of expression of CD44 on the cell surface was positive and reactions were mainly localized in cell membranes. Adhesion of trabecular meshwork cells to hyaluronate was inhibited by CD44-specific antisense oligonucleotide. Conclusions: Cultured human trabecular meshwork cells express CD44. CD44 may play a role in pathogenesis of primary open angle glaucoma. Eye Science 2004;20:52-56.展开更多
Purpose:To observe Fas expression change of cultured human retinal pigment epithelial (RPE) cells by IL-1β and TNF-α.Methods:With flow cytometry, immunohischemistry, and color imaging system, Fas expressions by expo...Purpose:To observe Fas expression change of cultured human retinal pigment epithelial (RPE) cells by IL-1β and TNF-α.Methods:With flow cytometry, immunohischemistry, and color imaging system, Fas expressions by exposure to IL-1β and/or TNF-α were measured.Results:The gray degree values of Fas expression were 67.5±6.1 in IL-1β+TNF-α-treated group, 80.1±9.2 in IL-1β-treated group, and 70.4±6.4 in TNF-α-treated group, respectively. There were significant differences (P < 0.005) compared with control group (107.0±10.2). Flow cytometry showed that 15.0% cultured human RPE cells expressed Fas. Fas-positive in IL-1β, TNF-α, and IL-1β+TNF-α-treated groups expressed was 28.1%, 34.5%, and 65.2%, respectively. Conclusion:IL-1β, TNF-α, and combining both of them can up-regulate Fas protein expression, which may contribute to more Fas (+) cells in proliferative vitreoretinopathy (PVR). Minimizing this process by means of inducing apoptosis of Fas (+) proliferative cells of Fas/FasL pathway is a future preventive and therapeutic possibility for PVR. Eye Science 2004;20:39-41.展开更多
Purpose: Dexamethasone(DEX) was tested for its ability to modulate human retinal pigment epithelium (hRPE) cell proliferation in cell culture.Methods: DEX in different concentrations was added to cultured hRPE cells. ...Purpose: Dexamethasone(DEX) was tested for its ability to modulate human retinal pigment epithelium (hRPE) cell proliferation in cell culture.Methods: DEX in different concentrations was added to cultured hRPE cells. The effects were measured with MTT method, 3H-thymidine(3H-TdR) incorporation and flow cytometw.Results: DEX increased survival rate and DNA synthesis from 32 mg/L to 320 mg/L under hRPE culture conditions, but paradoxically reduced them at 1 000 mg/L and 3 200 mg/L in dose and time dependent fashion by both MTT assay and 3 H-TdR incorporation. The cell numbers in S phase and G2/M phase increased 28.32 % at DEX concentration 320 mg/L, in contrast, reduced 41.84 % at 1 000 mg/L.Conclusion: DEX increased proliferation from 32 mg/L to 320 mg/L, and inhibited proliferation at concentrations greater than 320 mg/L. The inhibiting effect of DEX may happen in s phase and G2/M phase. Eye Sciernce 2001; 17:27 ~ 30.展开更多
Purpose: To explore the effect of external approach microsurgery in retinal dialysis.Methods: Consecutive 30 eyes of 28 patients with retinal dialysis were enrolled for this study. The progresses of the external appro...Purpose: To explore the effect of external approach microsurgery in retinal dialysis.Methods: Consecutive 30 eyes of 28 patients with retinal dialysis were enrolled for this study. The progresses of the external approach microsurgery were following. Under the surgical microscopy, the preplacement of mattress sutures for buckling and/or encircling following retrobulbar anesthesia and scleral exposure, draining subretinal fluid, the cryotherapy of retinal breaks, checking the position of breaks on scleral buckle and gases injection were performed in turn.Results: After drainage of subretinal fluid, with scleral depression cryotherapy reaction around breaks could be observed clearly under the microscopy. All breaks were located on anterior slope of the buckle. Intraoperative complications were mild subretinal hemorrhage at drainage site and corneal epithelium exfoliation in 3 eyes, respectively.Postoperative complications were mainly secondary glaucoma and retinal redetachment.The one-operation reattachmentl rate was 96.7% (29 eyes), and the final reattachment rate was 100% after one eye had a second external approach microsurgery. The postoperative vision acuity (VA) was significantly better than the preoperative VA (X2=9.529, P< 0.01).Conclusion: External approach microsurgery has favourable effect on the surgery of retinal dialysis.展开更多
Purpose: To study the expression of brain-derived neurotrophic factor (BDNF) in the rat retina delivered by adenovirus. Methods: Adenovirus with BDNF gene was injected into the vitreous. Gene expression was detected b...Purpose: To study the expression of brain-derived neurotrophic factor (BDNF) in the rat retina delivered by adenovirus. Methods: Adenovirus with BDNF gene was injected into the vitreous. Gene expression was detected by immunofluorescence staining,and quantitative analysis was performed after injury and transfection by Enzyme-linked immunosorbent assay (ELISA). Results: The positive cells can be seen on the 3rd day and last 4 weeks by immunofluore-scence staining. Positive cells in the control group were fewer than those in the transfection group or the fluorescence intensity was lower at every time point. Quantitative analysis showed that the expression of BDNF groups was higher than that of the control group at every time point(P < 0.01), and that of the injured group without transfection was higher than that of the control group on the 3rd day and the 7th day (P < 0.01).Conclusion: Efficient and stable transfer of BDNF gene could be achieved by adenovirus delivery into the retina of rats. Injury can promote the expression of BDNF in early period.展开更多
Purpose: To investigate the expression of matrix metalloproteinases (MMPs) in choroidal neovascular membranes with age-related macular degeneration (AMD).Methods: Seventeen choroidal neovascular membranes surgically r...Purpose: To investigate the expression of matrix metalloproteinases (MMPs) in choroidal neovascular membranes with age-related macular degeneration (AMD).Methods: Seventeen choroidal neovascular membranes surgically removed from AMD patients with pars plana vitrectomy and subretinal membranes peeling were investigated. The expression of MMP-2 and MMP-9 was determined with immunohistochemical technique. Results: Immunohistochemistry staining in choroidal neovascular membranes for MMP-2 and MMP-9 was observed in 17 specimens. There was no detective of MMP-2 and MMP-9 in normal retinas. Conclusions: MMP-2 and MMP-9 were found in choroidal neovascular membranes, may degrade the Bruch membrane and be associated with the perforation of new vessels into Bruch membrane, involving a basic pathogenic process of AMD.展开更多
Purpose: This is the first report of a bilateral nonischemic central retinal vein occlusionscombined with artery occlusions in a patient with acquired immune deficiency syndrome(AIDS). Methods: Case report. Results: A...Purpose: This is the first report of a bilateral nonischemic central retinal vein occlusionscombined with artery occlusions in a patient with acquired immune deficiency syndrome(AIDS). Methods: Case report. Results: A 22-year-old Chinese(male) with a positive human immunodeficiency virus(HIV) infection developed bilateral nonischemic central retinal vein occlusions combinedwith artery occlusions and severe vision loss. The manifestations of the fundus andfluorescein angiography were similar in both eyes.Conclusion: This case report provides the evidences that central retinal vein and arteryocclusions are probably part of the spectrum of AIDS vascular diseases.展开更多
Purpose:To evaluate the potential for Mecobalamin as a neuroprotective agent in optic nerve crush injury.Methods: Twenty-four adult Sprague-Dawley rats were randomly divided into four groups. One group acted as normal...Purpose:To evaluate the potential for Mecobalamin as a neuroprotective agent in optic nerve crush injury.Methods: Twenty-four adult Sprague-Dawley rats were randomly divided into four groups. One group acted as normal controls, while in the other three groups the right eye was subjected to optic nerve crush injury. Of the three crush injury groups one group received no treatment, while the other two groups received intramuscular injections of VitaminB12 or Mecobalamin (10μg) immediately after crush injury and then every two days. All the rats were sacrificed one month post-treatment, and the eyes attached with optic nerves were removed for histology. The morphological changes of optic nerve axons and retinal ganglion cells (RGCs) were assessed under light microscope (LM) and transmission electromicroscope (TEM). The numbers of axons and RGCs were counted. Results: In this study we demonstrate the potential for Mecobalamin as a neuroprotective agent following optic nerve crush injury. We show here that the axons of optic nerves were loose in structure or destroyed. The mitochondria of the RGCs was swollen, and the Nissel body was less evident after the crush injury. Moreover, the number of axons and RGCs was significantly reduced (P < 0.001). However, these changes were less dramatic after the Mecobalamin-treatment. More axons and RGCs were remained in the group than those in the untreated injury group (P = 0.010 and 0.003 respectively), and those in the VitaminB12-treated group (P =0.037 and 0.035 respectively). More significantly, there were newly formed axons found in the Mecobalamin-treated group.Conclusions: Optic nerve crush injury in rats causes the loss of the axons and RGCs but this may be ameliorated by treatment with Mecobalamin.展开更多
Purpose:To report ophthalmologic and angiographic features of choroidal metastases from carcinoid tumor and analyze their common clinical manifestation.Methods:Ophthalmologic examinations and fundus fluorescein angiog...Purpose:To report ophthalmologic and angiographic features of choroidal metastases from carcinoid tumor and analyze their common clinical manifestation.Methods:Ophthalmologic examinations and fundus fluorescein angiography(FFA) were performed in 30 patients suffered from carcinoid tumor,and four patients diagnosed of breast cancer(2 cases),lung cancer(1 case)and maxillary sinus cancer(1 case)were confirmed with choroidal metastases.Results:Choroidal metastases were found as the initial manifestations of the malignant tumors on 2 patients whose initial and chief complaints were decreasing vision,theirfun鄄dusl esions were mainly presented in the posterir pole and FFA showed high density of fluorescence of the lesions.Conclusions:This study indicated choroidal metastasis might be the first sign of metastases for patients with cancer.For patients with unknown metastastic cancers,examinations of the choroids may be useful for diagnosis and prognosis.Eye Science 2004;20:15-18.展开更多
Purpose: To investiagate the effect of antisense oligonucleotides (ODN) to Nogo-A mRNA expression in oligodendrocytes and to establish the base for further research of repair of optic nerve injury.Method: (1)Oligodend...Purpose: To investiagate the effect of antisense oligonucleotides (ODN) to Nogo-A mRNA expression in oligodendrocytes and to establish the base for further research of repair of optic nerve injury.Method: (1)Oligodendrocytes were obtained by inoculating the optic nerve of newborn (2 days)rats and were identified with galactocerebroside(GC) antibody immunocytochemical stain. (2) In order to observe the effects of antisense ODN on cultured cells,we set up five groups, including the groups of three concentration of antisense Nogo-A ODN (2μM、5μM、l0μM),a group with the random sequence added to the medium and the control group. Reverse transcription-polymerase chain reaction(RT-PCR) was adopted to study the effects of ODN on the expression of Nogo-A in oligodendrocytes.Results: (1)Three days after inoculation, a few of round or fusiform shape cells migrated from optic nerve tissue; About 11 days later, the coverlips were completely covered by the cells;The cells identified with GC antibody immunocytochemical stain were positive cells.(2)The result of RT-PCR study showed that antisense Nogo-A ODN could significantly and specifically inhibit the expression of Nogo-A after 24 hours ( P< 0.01). Random sequence has no effect on Nogo-A expresson.Conclusion:Antisense Nogo-A ODN can effectively and specifically inhibit the expression of Nogo-A.展开更多
AIM: To investigate the association between GSTM1 and GSTT1 polymorphisms and the risk of hepatocellular carcinoma (HCC) in Chinese population. METHODS: Literature databases including PubMed, ISI web of science and ot...AIM: To investigate the association between GSTM1 and GSTT1 polymorphisms and the risk of hepatocellular carcinoma (HCC) in Chinese population. METHODS: Literature databases including PubMed, ISI web of science and other databases were searched.Pooled odds ratio (OR) and 95% CI were calculated using random- or fixed-effects model. Subgroup analysis and sensitivity analysis were also performed. RESULTS: Nineteen studies of GSTM1 (2660 cases and 4017 controls) and 16 studies of GSTT1 (2410 cases and 3669 controls) were included. The GSTM1/GSTT1 null genotypes were associated with increased risk of HCC in Chinese population (for GSTM1, OR = 1.487, 95% CI: 1.159 to 1.908, P = 0.002; for GSTT1, OR = 1.510, 95% CI: 1.236 to 1.845, P = 0.000). No publication bias was detected. In subgroup analysis, glutathione S-transferases polymorphisms were significantly associated with HCC risk among the subjects living in high-incidence areas, but not among the subjects living in low-incidence areas. CONCLUSION: The present meta-analysis suggests that GSTM1/GSTT1 null genotypes are associated with increased risk of HCC in Chinese population.展开更多
Objective: To investigate the effects of nerve growth factor(NGF)on proliferation and DNAthesis of cultured human fetal retinal pigment epithelium (RPE)cells in vitro.Methods: Primary culture and subculture of human f...Objective: To investigate the effects of nerve growth factor(NGF)on proliferation and DNAthesis of cultured human fetal retinal pigment epithelium (RPE)cells in vitro.Methods: Primary culture and subculture of human fetal retinal pigment epithelium cellswere established in vitro first. Cultured RPE cells were treated with NGF by variousconcentrations 0μg/L, 50μg/L, 100μg/L, 200μg/L and 300μg/L(final concentration)for 48 hs.After 48 hs, cells proliferation was measured with methyl thiazolyl tetrazolium(MTT)assay method and the amount of DNA was determined by the absorbance at 280nm of nucleic acid & protein analysis.Results: The A values of 100 μg/L, 200 μg/L, 300 μg/L NGF was(0. 213 7 ± 0. 23 3),(0. 218 8 ±0. 018 1), (0. 232 2 ±0. 016 4) as compared with(0. 189 7 ±0. 015 2) of Avalue of 0 μg/L NGF respectively, q value was 3.63,4.40, 6. 42 and P value was0. 015, 0. 000, 0. 000(q-test). The DNA concentrations of 100 μg/L, 200 μg/L, 300μg/L and 400 μg/L NGF was (981. 220 4 ± 123.535 7), (1 375. 848 4 ±244. 471 8),(1 658.707 1 ± 176. 938 1), (2 353.086 3 ±609. 906 4) μg/ml as compared with(666. 818 8 ± 141. 330 2) μg/ml of DNA concentration of 0 μg/L NGF respectively, qvalue was 3.63,8.20,11.47,19.46, P value was 0. 024,0. 000,0. 000,0. 000 (q-test).Conclusion: The data suggested that NGF could stimulate the proliferation and DNAsynthesis of cultured of hRPE cells in vitro in a dose-dependent manner.展开更多
Orbscan- anterior system was used for early diagnosis of keratoconus.4 8Eyes of 2 4 patients with suspicious keratoconus were examined by Orbscan- anterior system from Dec. 1999to Dec. 2 0 0 0 and followed up.The ...Orbscan- anterior system was used for early diagnosis of keratoconus.4 8Eyes of 2 4 patients with suspicious keratoconus were examined by Orbscan- anterior system from Dec. 1999to Dec. 2 0 0 0 and followed up.The values of Diff and anterior chamber depth (ACD) were recorded.Results indicated that values of Diff and ACD were increased in4 eyes of2 patients with keratoconus trend during follow- up.Taking advantage of Orbscan- anterior system to observe the values of Diff and ACD can early diagnose the sub- clinical keratoconus.The values of Diff and ACD can sensitively report the progression of keratoconus.展开更多
Purpose: To treat the Terrien marginal degeneration using lamellar keratoplasty with dried corneosclera and evaluate the clinical effect of this operation.Methods: The study included 78 cases with Terrien marginal deg...Purpose: To treat the Terrien marginal degeneration using lamellar keratoplasty with dried corneosclera and evaluate the clinical effect of this operation.Methods: The study included 78 cases with Terrien marginal degeneration who underwent the lamellar keratoplasty with dried corneosclera. Among them, 48 cases in experiment Ⅰ group and 30 cases in experiment Ⅱ group underwent lamellar keratoplasty with glycerin-preserved corneosclera and anhydrous calcium chloride-storeded corneosclera respectively. All the patients were followed-up from 6 months to 10 years. The control group consisted of 38 cases with Terrien marginal degeneration electively undergoing lamellar keratoplasty with fresh corneosclera simultaneously.Results: The area of the corneal thinning was mended and the eyeball was saved. The corneal astigmatism of the patients was corrected obviously.The patient’s vision was improved significantly. No recurrence and rejection were observed after the operation.There is no significant difference between the experiment group (experiment Ⅰ group or experiment Ⅱ group) and control group. (P > 0.05), despite longer duration of postoperative corneal edema in experiment group.Conclusions: The lamellar keratoplasty with dried corneosclera for treatment of Terrien marginal degeneration is a safe and effective operation. It is an alternative to lamellar keratoplasty with fresh corneosclera for treatment of Terrien marginal degeneration. It is suitable to be popularized in the local hospital of our country.展开更多
Purpose: To compare the differences of corneal topographies in myopic and hyperopic children and study the effect of Atropin on their changes.Methods: The refractive components of 136 eyes with different refractive co...Purpose: To compare the differences of corneal topographies in myopic and hyperopic children and study the effect of Atropin on their changes.Methods: The refractive components of 136 eyes with different refractive conditions were measured with A-Scan and their corneal topographies with and without cycloplegia were obtained respectively.Results: The mean corneal power of zones 3mm (MD3, P=0.031 ) and minor keratometer K2 (P=0.003) of myopia are greater than those of hyperopia without cycloplegia. MD3 (P=0.009) and Keratometer K1 (P = 0.025) increased in hyperopic eyes, while MD3(P=0.033), K1 (P = 0.035) and K2 (P = 0.002) decreased in myopic eyes significantly after cycloplegia. Similarly, the mean corneal power of zones 5mm (MD5) and 7mm (MD7) in myopic eyes decreased dramatically (P ≤ 0.001 ).Conclusions: The corneal power was found to be greater in myopia than that in hyperopia. The effect of Atropin on corneal shape of myopia and hyperopia was in the opposite direction.展开更多
Purpose:To study the densities and distribution of retinal ganglion cells(RGC)in adult rat retinae with flurogold(FG)labeling retogradely.Methods:FG was injected to the superior colliculi(SC)and dorsal lateral genicul...Purpose:To study the densities and distribution of retinal ganglion cells(RGC)in adult rat retinae with flurogold(FG)labeling retogradely.Methods:FG was injected to the superior colliculi(SC)and dorsal lateral geniculate nuclei(dLGN)in adult rats and the retinae were examined by fluorescence microscopy at various periods of time.Results;FG-labelled RGC were observed in the retina as early as 3days after application of FG.The labelled cells gradually increased in density,reached95%of the maximal number on days7and the maximal number on days30.The density of labelled cells was higher in the posterior pole than in the peripheral area.The fluorescence intensity in labelled cells maintained up to 60days.Conclusion:The FG retrograde labeling method is reliable and effective for quantity of RGC.Eye Science2000;16:29-33.展开更多
文摘Purpose:To investigate the dynamics of heat shock protein 72(HSP72) expression in retinal ganglion cells(RGCs) in rat model of acute glaucoma treated with heat stress or intraperitoneal injection of zinc sulfate.Methods:Twenty-seven male Wistar rats were used to make acute glaucoma models. Five others served as normal control. Acute glaucoma models were made by intracameral irrigation in the right eyes with balanced salt saline (BSS) at 102 mmHg for 2 hours. Nine model rats were killed at different intervals after intracameral irrigation without treatment,which served as damage control. Ten were treated with heat stress 40℃~42℃, and 8 were used for zinc sulfate administration 2 days posterior to intracameral irrigation. Treated model rats were sacrificed at designed intervals after treatment. Right eyes were enucleated immediately, and the retinas were dissected for Western blot.Results:No HSP72 was found in RGCs of normal Wistar rats. In damage control group, slight HSP72 was detected during 6~36 hours posterior to intracameral irrigation. HSP72 was detected significantly expressed in RGCs of both heat shock group and zinc sulfate group. But the dynamics of HSP72 production were quite different in these two treated groups. In heat shock group, HSP72 appeared at the sixth hour after treatment, and increased gradually until its peak production emerged at the 48th hour. HSP72 vanished 8 days later after treatment. In zinc sulfate group, HSP72 expression began 24 hours later after zinc administration, and reached its highest level at the 72th hour posterior to treatment. HSP72 expression then decreased slowly, and disappeared 21 days later after treatment. Conclusion:HSP72 can be induced in RGCs of rat acute glaucoma models with heat stress or zinc sulfate adddministration. But the dynamics of the HSP72 induction in those two groups were quite different. Eye Science 2004;20:30-33.
文摘Purpose: To investigate the effect of high dose methylprednisolone (MP) on retinal ganglion cells (RGC) apoptosis.Methods: One hundred and twenty six Wistar rats were divided into normal control, crush control and MP treatment groups. High dose MP and 0.9% sodium chloride were injected intravenously at 1 h after injury in the treatment group and crush control group, respectively. Apoptotic cells, Bcl-2 and Bax positive cells were located and calculated by terminal-deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling (TUNEL), immunohistochemical method and retinal whole-mount combined with section technique at day 4, 7, 14 after crush, respectively.Results: The TUNEL-labeling cells, Bcl-2 and Bax positive cells were detected in the ganglion cell layer of retina. Few apoptotic cells, a few of Bcl-2 and Bax positive cells were observed in normal control groups. The numbers of positive cells increased gradually after crush. The number of apoptotic cells and Bax positive cells in treatment groups was less than that in crush group significantly (P < 0.05), while the number of Bcl-2 positive cells in treatment group was more than that in crush group at day 4, 7 and 14 after crush significantly (P < 0.05) Conclusions: These results suggest that high dose MP can inhibit the apoptosis of RGC after optic nerve crush probably through up-regulation of Bcl-2 expression and down-regulation of Bax expression.
基金Supported by National Natural Science Foundation of China (30170997)
文摘Purpose: To investigate the effects of culture medium of human amniotic membrane (AM) on corneal neovascularization (CNV) induced by basic fibroblast growth factor (bFGF) in mice.Methods: Culture medium of amniotic membrane was prepared by cultivating AM (with epithelium side up) in EGM basic medium for 3 days, and was collected separately to three groups, e.g. Control (EGM only), AM with epithelium (AM) and AM without epithelium (De-AM). Corneal neovascularization was induced in mice by using micropocket assay with Hydron polymer pellets containing 100 ng bFGF. Migration and proliferation of human umbilical cord vein endothelial cells (HUVEC) were performed in Boyden chambers and by using the CyQUANT fluorescence binding assay respectively.The levels of tissue inhibitors of metalloproteinase 1 and 2 (TIMP1, TIMP2) in culture medium were determined by ELISA assay.Results: CNV induced by bFGF was significantly suppressed by culture medium of amniotic membrane. When the medium was applied as an eyedrop 4 times a day for 7 days,the area of CNV was (2.48±0.76) mm2,(0.64±0.52) mm2 and (1.96±0.65) mm2 incontrol, AM and De-AM group respectively. The migration and proliferation of HUVEC were strongly inhibited by culture medium of AM with epithelium, while the De-AM had no effect on the migration of HUVEC cells. The high level of TIMP2 was found in AM group, but not in De-AM group, while there was no difference in the amount of TIMP1 in medium among three groups.Conclusion: Culture medium of amniotic membrane significantly suppresses the corneal nevovascularization induced by bFGF. The mechanism of which at least in part is that high level of TIMP2 protein secreted or released into the culture medium of AM and inhibition of migration and growth of vascular endothelial cells.
文摘Purpose:To determine whether cultured human trabecular meshwork cells express CD44 and to discuss their possible relationship with primary open angle glaucoma.Methods:Human trabecular meshwork cells were cultured in DMEM/F12 media. Total RNAs from the cells were extracted with Trizol reagent. Messenger RNA expression of CD44 in human trabecular meshwork cells was examined by using reverse transcriptase-polymerase chain reaction ( RT-PCR ) analysis. Expression of CD44 was confirmed by Western-blotting and immunofluorescent microscopy. Effect of CD44-specific antisense oligonucleotide on adhesion of trabecular meshwork cells to hyaluronate was determined by MTT assay. Results:A single RT-PCR product whose size was 471bp was obtained. A band about 80kD was stained by Western-blot. Immunofluorescent examination of expression of CD44 on the cell surface was positive and reactions were mainly localized in cell membranes. Adhesion of trabecular meshwork cells to hyaluronate was inhibited by CD44-specific antisense oligonucleotide. Conclusions: Cultured human trabecular meshwork cells express CD44. CD44 may play a role in pathogenesis of primary open angle glaucoma. Eye Science 2004;20:52-56.
文摘Purpose:To observe Fas expression change of cultured human retinal pigment epithelial (RPE) cells by IL-1β and TNF-α.Methods:With flow cytometry, immunohischemistry, and color imaging system, Fas expressions by exposure to IL-1β and/or TNF-α were measured.Results:The gray degree values of Fas expression were 67.5±6.1 in IL-1β+TNF-α-treated group, 80.1±9.2 in IL-1β-treated group, and 70.4±6.4 in TNF-α-treated group, respectively. There were significant differences (P < 0.005) compared with control group (107.0±10.2). Flow cytometry showed that 15.0% cultured human RPE cells expressed Fas. Fas-positive in IL-1β, TNF-α, and IL-1β+TNF-α-treated groups expressed was 28.1%, 34.5%, and 65.2%, respectively. Conclusion:IL-1β, TNF-α, and combining both of them can up-regulate Fas protein expression, which may contribute to more Fas (+) cells in proliferative vitreoretinopathy (PVR). Minimizing this process by means of inducing apoptosis of Fas (+) proliferative cells of Fas/FasL pathway is a future preventive and therapeutic possibility for PVR. Eye Science 2004;20:39-41.
文摘Purpose: Dexamethasone(DEX) was tested for its ability to modulate human retinal pigment epithelium (hRPE) cell proliferation in cell culture.Methods: DEX in different concentrations was added to cultured hRPE cells. The effects were measured with MTT method, 3H-thymidine(3H-TdR) incorporation and flow cytometw.Results: DEX increased survival rate and DNA synthesis from 32 mg/L to 320 mg/L under hRPE culture conditions, but paradoxically reduced them at 1 000 mg/L and 3 200 mg/L in dose and time dependent fashion by both MTT assay and 3 H-TdR incorporation. The cell numbers in S phase and G2/M phase increased 28.32 % at DEX concentration 320 mg/L, in contrast, reduced 41.84 % at 1 000 mg/L.Conclusion: DEX increased proliferation from 32 mg/L to 320 mg/L, and inhibited proliferation at concentrations greater than 320 mg/L. The inhibiting effect of DEX may happen in s phase and G2/M phase. Eye Sciernce 2001; 17:27 ~ 30.
文摘Purpose: To explore the effect of external approach microsurgery in retinal dialysis.Methods: Consecutive 30 eyes of 28 patients with retinal dialysis were enrolled for this study. The progresses of the external approach microsurgery were following. Under the surgical microscopy, the preplacement of mattress sutures for buckling and/or encircling following retrobulbar anesthesia and scleral exposure, draining subretinal fluid, the cryotherapy of retinal breaks, checking the position of breaks on scleral buckle and gases injection were performed in turn.Results: After drainage of subretinal fluid, with scleral depression cryotherapy reaction around breaks could be observed clearly under the microscopy. All breaks were located on anterior slope of the buckle. Intraoperative complications were mild subretinal hemorrhage at drainage site and corneal epithelium exfoliation in 3 eyes, respectively.Postoperative complications were mainly secondary glaucoma and retinal redetachment.The one-operation reattachmentl rate was 96.7% (29 eyes), and the final reattachment rate was 100% after one eye had a second external approach microsurgery. The postoperative vision acuity (VA) was significantly better than the preoperative VA (X2=9.529, P< 0.01).Conclusion: External approach microsurgery has favourable effect on the surgery of retinal dialysis.
文摘Purpose: To study the expression of brain-derived neurotrophic factor (BDNF) in the rat retina delivered by adenovirus. Methods: Adenovirus with BDNF gene was injected into the vitreous. Gene expression was detected by immunofluorescence staining,and quantitative analysis was performed after injury and transfection by Enzyme-linked immunosorbent assay (ELISA). Results: The positive cells can be seen on the 3rd day and last 4 weeks by immunofluore-scence staining. Positive cells in the control group were fewer than those in the transfection group or the fluorescence intensity was lower at every time point. Quantitative analysis showed that the expression of BDNF groups was higher than that of the control group at every time point(P < 0.01), and that of the injured group without transfection was higher than that of the control group on the 3rd day and the 7th day (P < 0.01).Conclusion: Efficient and stable transfer of BDNF gene could be achieved by adenovirus delivery into the retina of rats. Injury can promote the expression of BDNF in early period.
文摘Purpose: To investigate the expression of matrix metalloproteinases (MMPs) in choroidal neovascular membranes with age-related macular degeneration (AMD).Methods: Seventeen choroidal neovascular membranes surgically removed from AMD patients with pars plana vitrectomy and subretinal membranes peeling were investigated. The expression of MMP-2 and MMP-9 was determined with immunohistochemical technique. Results: Immunohistochemistry staining in choroidal neovascular membranes for MMP-2 and MMP-9 was observed in 17 specimens. There was no detective of MMP-2 and MMP-9 in normal retinas. Conclusions: MMP-2 and MMP-9 were found in choroidal neovascular membranes, may degrade the Bruch membrane and be associated with the perforation of new vessels into Bruch membrane, involving a basic pathogenic process of AMD.
文摘Purpose: This is the first report of a bilateral nonischemic central retinal vein occlusionscombined with artery occlusions in a patient with acquired immune deficiency syndrome(AIDS). Methods: Case report. Results: A 22-year-old Chinese(male) with a positive human immunodeficiency virus(HIV) infection developed bilateral nonischemic central retinal vein occlusions combinedwith artery occlusions and severe vision loss. The manifestations of the fundus andfluorescein angiography were similar in both eyes.Conclusion: This case report provides the evidences that central retinal vein and arteryocclusions are probably part of the spectrum of AIDS vascular diseases.
文摘Purpose:To evaluate the potential for Mecobalamin as a neuroprotective agent in optic nerve crush injury.Methods: Twenty-four adult Sprague-Dawley rats were randomly divided into four groups. One group acted as normal controls, while in the other three groups the right eye was subjected to optic nerve crush injury. Of the three crush injury groups one group received no treatment, while the other two groups received intramuscular injections of VitaminB12 or Mecobalamin (10μg) immediately after crush injury and then every two days. All the rats were sacrificed one month post-treatment, and the eyes attached with optic nerves were removed for histology. The morphological changes of optic nerve axons and retinal ganglion cells (RGCs) were assessed under light microscope (LM) and transmission electromicroscope (TEM). The numbers of axons and RGCs were counted. Results: In this study we demonstrate the potential for Mecobalamin as a neuroprotective agent following optic nerve crush injury. We show here that the axons of optic nerves were loose in structure or destroyed. The mitochondria of the RGCs was swollen, and the Nissel body was less evident after the crush injury. Moreover, the number of axons and RGCs was significantly reduced (P < 0.001). However, these changes were less dramatic after the Mecobalamin-treatment. More axons and RGCs were remained in the group than those in the untreated injury group (P = 0.010 and 0.003 respectively), and those in the VitaminB12-treated group (P =0.037 and 0.035 respectively). More significantly, there were newly formed axons found in the Mecobalamin-treated group.Conclusions: Optic nerve crush injury in rats causes the loss of the axons and RGCs but this may be ameliorated by treatment with Mecobalamin.
文摘Purpose:To report ophthalmologic and angiographic features of choroidal metastases from carcinoid tumor and analyze their common clinical manifestation.Methods:Ophthalmologic examinations and fundus fluorescein angiography(FFA) were performed in 30 patients suffered from carcinoid tumor,and four patients diagnosed of breast cancer(2 cases),lung cancer(1 case)and maxillary sinus cancer(1 case)were confirmed with choroidal metastases.Results:Choroidal metastases were found as the initial manifestations of the malignant tumors on 2 patients whose initial and chief complaints were decreasing vision,theirfun鄄dusl esions were mainly presented in the posterir pole and FFA showed high density of fluorescence of the lesions.Conclusions:This study indicated choroidal metastasis might be the first sign of metastases for patients with cancer.For patients with unknown metastastic cancers,examinations of the choroids may be useful for diagnosis and prognosis.Eye Science 2004;20:15-18.
基金This work was supported by the Natural Science National Foundation of China (grant number 30270458)the Application Basic Research Foundation of Chongqing Scientific Committee (grant number 41A1152C)the Fifteenth Medicine and Health Scientific Researc
文摘Purpose: To investiagate the effect of antisense oligonucleotides (ODN) to Nogo-A mRNA expression in oligodendrocytes and to establish the base for further research of repair of optic nerve injury.Method: (1)Oligodendrocytes were obtained by inoculating the optic nerve of newborn (2 days)rats and were identified with galactocerebroside(GC) antibody immunocytochemical stain. (2) In order to observe the effects of antisense ODN on cultured cells,we set up five groups, including the groups of three concentration of antisense Nogo-A ODN (2μM、5μM、l0μM),a group with the random sequence added to the medium and the control group. Reverse transcription-polymerase chain reaction(RT-PCR) was adopted to study the effects of ODN on the expression of Nogo-A in oligodendrocytes.Results: (1)Three days after inoculation, a few of round or fusiform shape cells migrated from optic nerve tissue; About 11 days later, the coverlips were completely covered by the cells;The cells identified with GC antibody immunocytochemical stain were positive cells.(2)The result of RT-PCR study showed that antisense Nogo-A ODN could significantly and specifically inhibit the expression of Nogo-A after 24 hours ( P< 0.01). Random sequence has no effect on Nogo-A expresson.Conclusion:Antisense Nogo-A ODN can effectively and specifically inhibit the expression of Nogo-A.
基金Supported by (partially) The Heilongjiang Provincial Health Department No. 2009-201+1 种基金the Administration of Traditional Chinese Medicine of Heilongjiang Province No. ZHY10-293
文摘AIM: To investigate the association between GSTM1 and GSTT1 polymorphisms and the risk of hepatocellular carcinoma (HCC) in Chinese population. METHODS: Literature databases including PubMed, ISI web of science and other databases were searched.Pooled odds ratio (OR) and 95% CI were calculated using random- or fixed-effects model. Subgroup analysis and sensitivity analysis were also performed. RESULTS: Nineteen studies of GSTM1 (2660 cases and 4017 controls) and 16 studies of GSTT1 (2410 cases and 3669 controls) were included. The GSTM1/GSTT1 null genotypes were associated with increased risk of HCC in Chinese population (for GSTM1, OR = 1.487, 95% CI: 1.159 to 1.908, P = 0.002; for GSTT1, OR = 1.510, 95% CI: 1.236 to 1.845, P = 0.000). No publication bias was detected. In subgroup analysis, glutathione S-transferases polymorphisms were significantly associated with HCC risk among the subjects living in high-incidence areas, but not among the subjects living in low-incidence areas. CONCLUSION: The present meta-analysis suggests that GSTM1/GSTT1 null genotypes are associated with increased risk of HCC in Chinese population.
文摘Objective: To investigate the effects of nerve growth factor(NGF)on proliferation and DNAthesis of cultured human fetal retinal pigment epithelium (RPE)cells in vitro.Methods: Primary culture and subculture of human fetal retinal pigment epithelium cellswere established in vitro first. Cultured RPE cells were treated with NGF by variousconcentrations 0μg/L, 50μg/L, 100μg/L, 200μg/L and 300μg/L(final concentration)for 48 hs.After 48 hs, cells proliferation was measured with methyl thiazolyl tetrazolium(MTT)assay method and the amount of DNA was determined by the absorbance at 280nm of nucleic acid & protein analysis.Results: The A values of 100 μg/L, 200 μg/L, 300 μg/L NGF was(0. 213 7 ± 0. 23 3),(0. 218 8 ±0. 018 1), (0. 232 2 ±0. 016 4) as compared with(0. 189 7 ±0. 015 2) of Avalue of 0 μg/L NGF respectively, q value was 3.63,4.40, 6. 42 and P value was0. 015, 0. 000, 0. 000(q-test). The DNA concentrations of 100 μg/L, 200 μg/L, 300μg/L and 400 μg/L NGF was (981. 220 4 ± 123.535 7), (1 375. 848 4 ±244. 471 8),(1 658.707 1 ± 176. 938 1), (2 353.086 3 ±609. 906 4) μg/ml as compared with(666. 818 8 ± 141. 330 2) μg/ml of DNA concentration of 0 μg/L NGF respectively, qvalue was 3.63,8.20,11.47,19.46, P value was 0. 024,0. 000,0. 000,0. 000 (q-test).Conclusion: The data suggested that NGF could stimulate the proliferation and DNAsynthesis of cultured of hRPE cells in vitro in a dose-dependent manner.
文摘Orbscan- anterior system was used for early diagnosis of keratoconus.4 8Eyes of 2 4 patients with suspicious keratoconus were examined by Orbscan- anterior system from Dec. 1999to Dec. 2 0 0 0 and followed up.The values of Diff and anterior chamber depth (ACD) were recorded.Results indicated that values of Diff and ACD were increased in4 eyes of2 patients with keratoconus trend during follow- up.Taking advantage of Orbscan- anterior system to observe the values of Diff and ACD can early diagnose the sub- clinical keratoconus.The values of Diff and ACD can sensitively report the progression of keratoconus.
文摘Purpose: To treat the Terrien marginal degeneration using lamellar keratoplasty with dried corneosclera and evaluate the clinical effect of this operation.Methods: The study included 78 cases with Terrien marginal degeneration who underwent the lamellar keratoplasty with dried corneosclera. Among them, 48 cases in experiment Ⅰ group and 30 cases in experiment Ⅱ group underwent lamellar keratoplasty with glycerin-preserved corneosclera and anhydrous calcium chloride-storeded corneosclera respectively. All the patients were followed-up from 6 months to 10 years. The control group consisted of 38 cases with Terrien marginal degeneration electively undergoing lamellar keratoplasty with fresh corneosclera simultaneously.Results: The area of the corneal thinning was mended and the eyeball was saved. The corneal astigmatism of the patients was corrected obviously.The patient’s vision was improved significantly. No recurrence and rejection were observed after the operation.There is no significant difference between the experiment group (experiment Ⅰ group or experiment Ⅱ group) and control group. (P > 0.05), despite longer duration of postoperative corneal edema in experiment group.Conclusions: The lamellar keratoplasty with dried corneosclera for treatment of Terrien marginal degeneration is a safe and effective operation. It is an alternative to lamellar keratoplasty with fresh corneosclera for treatment of Terrien marginal degeneration. It is suitable to be popularized in the local hospital of our country.
文摘Purpose: To compare the differences of corneal topographies in myopic and hyperopic children and study the effect of Atropin on their changes.Methods: The refractive components of 136 eyes with different refractive conditions were measured with A-Scan and their corneal topographies with and without cycloplegia were obtained respectively.Results: The mean corneal power of zones 3mm (MD3, P=0.031 ) and minor keratometer K2 (P=0.003) of myopia are greater than those of hyperopia without cycloplegia. MD3 (P=0.009) and Keratometer K1 (P = 0.025) increased in hyperopic eyes, while MD3(P=0.033), K1 (P = 0.035) and K2 (P = 0.002) decreased in myopic eyes significantly after cycloplegia. Similarly, the mean corneal power of zones 5mm (MD5) and 7mm (MD7) in myopic eyes decreased dramatically (P ≤ 0.001 ).Conclusions: The corneal power was found to be greater in myopia than that in hyperopia. The effect of Atropin on corneal shape of myopia and hyperopia was in the opposite direction.
基金The project was supported by the National Natural Science Foundation of China (No.39900076)
文摘Purpose:To study the densities and distribution of retinal ganglion cells(RGC)in adult rat retinae with flurogold(FG)labeling retogradely.Methods:FG was injected to the superior colliculi(SC)and dorsal lateral geniculate nuclei(dLGN)in adult rats and the retinae were examined by fluorescence microscopy at various periods of time.Results;FG-labelled RGC were observed in the retina as early as 3days after application of FG.The labelled cells gradually increased in density,reached95%of the maximal number on days7and the maximal number on days30.The density of labelled cells was higher in the posterior pole than in the peripheral area.The fluorescence intensity in labelled cells maintained up to 60days.Conclusion:The FG retrograde labeling method is reliable and effective for quantity of RGC.Eye Science2000;16:29-33.