OBJECUIVE: To observe the therapeutic efficacy of gamma;-knife/lymphokine activated killing cells (LAK)in chorold malignant melanoma (CMM). METHODS:Five cases of CMM had keen treated by retrobulbar injection of LAK cells and gamma;-knife irradiation at multiple sites.Ophthalmologic,imageologic, fundus fluorescein angiographic and T lymphocyte subset examinations were done before and after treatment. Tile follow-up period of this series of cases was 6-24 months. RESUILS:Thc CMM of 4 in 5 treated cases became atrophic and withered up clinically after gamma;-kinfe/LAK therapy. Among the 4 cases,2 of them had been followed up for more than 2 years,and the other 2 for 20 and 14 months respectively. The tumor of the 5th patient wko was followed up for 6 months after treatment,reduced to 3/5 of the original size,and no blood flow was found within thee tumor mass under the clinical examination. CONCLUSION :The gamma;-knife/LAK therapy was effective in treating CMM in saving the affected eye from being enucleated. Chin J Ocul Fundus Dis,1997,13: 96- 98）
ObjectiveTo observe the protective effect of Zhicao Tea Mixture on Müller cells and the expression of inflammatory factors in mice with diabetic retinopathy.MethodsSeventy-five C57BL/6J mice were randomly divided into the normal control group, diabetes mellitus (DM) group, low concentrations group, medium concentrations group and high concentrations group, with 16 mice in each group. The diabetes model of mice in all groups except the normal control group were established by intraperitoneal injection of STZ (60 mg/kg). Four weeks after the successful modeling, the Zhicao Tea Mixture with low (30 ml/kg), medium (60 ml/kg) and high concentrations (120 ml/kg) were respectively administered by gavage. Weight and blood glucose of mice in each group were measured every two weeks. After 8 weeks, Western blot method was used to detect the mice retina Müller cells activation marker gelatinous fibrous acidic protein (GFAP). Immunofluorescence was performed to detect the expression GFAP and glutamine synthetase (GS). Real-time quantitative PCR (RT-qPCR) and ELISA were used to determine the mRNA and protein expression levels of mouse retinal VEGF, TNF-α, IL-1β and IL-6 respectively.ResultsThe weight of mice in the DM group was lower than that of the normal control group, and the blood glucose was increased. Zhicao Tea Mixture had no effect on the weight of DM mice, but had a significant hypoglycemic effect. The GFAP expression (t=38.318, P＜0.001) in the retina of mice in the DM group was increased and GS expression (t=29.737, P＜0.001) was decreased compared with the control group. The GFAP expression (t=13.677, 19.387, 16.305; P＜0.05) in the retina of mice in the low, medium and high concentrations group were decreased and GS expression (t=5.170, 19.399, 6.705; P＜0.05) were increased compared with the DM group. The expressions of retinal inflammatory factors VEGF, TNF-α, IL-1β and IL-6 in DM group all increased, while the expressions of the above-mentioned inflammatory factors in the retina of mice decreased in the low, medium and high concentrations group.ConclusionZhicao Tea Mixture can decrease the blood glucose of DM mice and reduces the diabetic retinal inflammatory response.
Tumor-derived exosomes play a role in helping tumor cells with escape from immune surveillance, and it may also activate tumor-specific immune responses to eradicate tumor cells. Tumor cells release exosomes with major histocompatibility complex molecules and antigenic peptides on the surface membranes, which can induce dendritic cells (DC) and cytokine-induced killer (CIK) cells in vitro to produce the tumor antigen-specific T cells, and the obtained DC-CIK cells have a dual antitumor function with specificity and non specificity. This provides a new method for the treatment of cancers. This review briefly summarized the latest progress of adoptive immunotherapy with exosomes and DC-CIK.
Ischemic retinopathy, resulting in multiple lesions like microvasculature damage, inflammation and neovascularization, is a major contributor of vision damage. In these pathological changes, retinal glia cannot be ignored in the development of retinopathy. They constitute a highly versatile population that interacts with various cells to maintain homeostasis and limit disease. Therefore, glial activation and gliosis are strikingly ubiquitous responses to almost every form of retinal disease. Both of microglial cells and Müller cells are major intrinsic retinal glial cells and they are in close relationship, which means they can influence each other, make joint action or even become interdependent. They exhibit morphological and functional changes to have an impact on degree of retinal injury through different responses, which mediated by glial cells are important not only for course of disease progression, but also for the maintenance of neuronal and photoreceptor survival. Thus, defining the mechanisms that underlie communications between microglial cells and Müller cells could enable the development of more selective therapeutic targets, with great potential clinical applications.
Aminoadipic acid(AAA) is known to damage retinal glia cells primarily when it is given to animals intravitreally. The present study is to demonstrate marked increase of retinal susceptibility to photic damage following administration of sub-thres-hold doses of this agent to albino rats. Right eyes were intravitreally injected with 10 ?l of 10 mM AAA, a dose which caused transient swelling of Muller cell nucleiimmediately after treatment, and total recovery by 24 hours. These rats were exposed to fluorescent light at 150 f.c. for one hour three days after injection. The left eyes were injected with the same amount of physiologic saline solution and exposed to light with an identical time schedule. The animals were killed at the 24th hour,third and seventh day, following light exposure. Cytologic changes in the retinae of both eyes were compared light microscopically. The light exposed left eyes showed mild disorganization of photoreceptor outer segements. Usually this change disappeared by the seventh day. AAA-injected right eyes showed marked destruction in the photoreceptor cell layer. The change in the photoreceptor cells was progressive and disappearance of outer segments and degeneration of numerous nuclei occurred during the following period. （Chin J Ocul Fundus Dis,1992,8:17-19）
ObjectiveTo observe the expression of probucol on high glucose-induced specificity protein 1(SP1), kelchlike ECH associated protein1 (Keap1), NF-E2-related factor 2 (Nrf2) and glutamate-cysteine ligase catalytic (GCLC) in the cultured human müller cells and preliminary study the antioxidation of the probucol on müller cells.MethodsPrimary cultured human müller cells were randomly divided into four groups: normoglycaemia group (5.5 mmol/L glucose), normoglycaemia with probucol group (5.5 mmol/L glucose+100 μmol/L probucol), hyperglycemia group (25.0 mmol/L glucose), hyperglycemia with probucol group (25.0 mmol/L glucose + 100 μmol/L probucol). Immunofluorescence staining was used to assess distribution of SP1, Keap1, Nrf2, GCLC in human Müller cells. SP1, Keap1, Nrf2 and GCLC messenger RNA (mRNA) expression was evaluated by quantitative real-time RT-PCR (qRT-PCR). Independent sample t test was used to compare the data between the two groups.ResultsAll müller cells expressed glutamine synthetase (＞95%), which confirmed the cultured cells in vitro were the purification of generations of müller cells. The expressions of SP1, Keap1, Nrf2, and GCLC protein were positive in human müller cells. qRT-PCR indicated that SP1 (t=28.30, P＜0.000), Keap1 (t=5.369, P=0.006), and Nrf2 (t=10.59, P=0.001) mRNA in the hyperglycemia group increased obviously compared with the normoglycaemia group; GCLC (t=4.633, P=0.010) mRNA in the hyperglycemia group decreased significantly compared with the normoglycaemia group. However, SP1 (t=12.60, P=0.000) and Keap1 (t=4.076, P=0.015) in the hyperglycemia with probucol group decreased significantly compared with the hyperglycemia group; Nrf2 (t=12.90, P=0.000) and GCLC (t=15.96, P＜0.000) mRNA in the hyperglycemia with probucol group increased obviously compared with with the hyperglycemia group.ConclusionProbucol plays an antioxidant role by inhibiting the expression of SP1, Keap1 and up-regulating the expression of Nrf2, GCLC in müller cells induced by high glucose.
ObjectiveTo observe the effect of tert-Butylhydroquinone (tBHQ) on the expression of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase (HO)-1 and phosphatidylinositol 3-kinase (PI3K) in high glucose cultured retinal Müller cells; and to investigate the anti-oxidative stress and anti-apoptotic effects of tBHQ.MethodsRetinal Müller cells were divided into normal glucose group (5.5 mmol/L, N group), high glucose group (45 mmol/L, HG group) and tBHQ intervention group (HG+tBHQ group). After retinal Müller cells were cultured with high glucose for 48 hours, the pretreatment with tBHQ (20 μmol/L) induced the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and HO-1. The Müller cells were identified by immunofluorescence staining. The expressions of Nrf2, HO-1, PI3K, B-cell lymphoma-2 (Bcl-2) and Bax were detected by Western blot and real-time fluorescence quantitative PCR. Flow cytometry was used to detect the apoptosis of retinal Müller cells in rats.ResultsMüller cytoplasm and nucleus GS showed strong positive, large cell body, abundant cytoplasm, uniform green fluorescence; nuclear DAPI staining round or oval, clear boundary. The expression of Nrf2 protein (t=4.114, P=0.006), HO-1 protein (t=9.275, P=0.000), Nrf2 mRNA (t=7.292, P=0.000) and HO-1 mRNA (t=15.014, P=0.000) in the HG group were higher than those in the N group. The expressions of Nrf2 protein (t=7.847, P=0.000) ,HO-1 protein (t=7.947, P=0.000), PI3K protein (t=5.397, P=0.002), Bcl-2 protein (t=6.825, P=0.000), Nrf2 mRNA (t=18.046, P=0.000), HO-1 mRNA (t=39.458, P=0.000), PI3K mRNA (t=4.979, P=0.003) and Bcl-2 mRNA (t=9.535, P=0.000) in the HG+tBHQ group were significantly higher than those in the HG group. The protein and mRNA expressions of Bax protein in the HG+tBHQ group were significantly lower than those in the HG group (t=14.998, 16.520; P=0.000, 0.000). Flow cytometry showed that the apoptosis rate of Müller cells in the HG group was significantly higher than that in the N group (t=39.905, P=0.000). The apoptosis rate of Müller cells in the HG+tBHQ group was significantly lower than that in the HG group (t=21.083, P=0.000).ConclusiontBHQ can inhibit the apoptosis of retinal Müller cells by up-regulating the expression of Nrf2, HO-1 and PI3K.
Objective To investigate the effect of hypoxia on expressions of erythropoietin（EPO）mRNA and protein in retinal Muuml;ller cells cultured in vitro. Methods Retina tissues from the new-born Wistar rats were dissected into cell suspension after digested by pancreatin.Muuml;ller cells were separated and purified by mechanical concussion and blowing and striking method.The expression of EPO mRNA and protein under the condition of hypoxia was detected by semi-quantitative reverse transcriptase（RT）-polymerase chain reaction（PCR）and immunocytochemical method. Results Retinal Muuml;ller cells were cultured successfully，95% of which were positively stained by glial fibrillary acidic protein（GFAP）.Positively stained EPO protein was located in the cytoplasm and protuberance.The expression of EPO mRNA and protein was faint in the normal retinal Muuml;ller cells，but increased obviously and time-dependently after hypoxia. Conclusion Expression of EPO mRNA and protein increases in Muuml;ller cells after hypoxia，which may be one of the protective factors for the nerves in anoxic retinopathy. (Chin J Ocul Fundus Dis, 2006, 22: 196-199)
Objective To investigate the effect of long-term intraocular retention of domestic perfluorocarbon liquid (PFCL) on morphology and histology of ocular tissues. Methods A total of 18 New-Zealand rabbits were randomly divided into 3 experimental groups, whose left eyes underwent intraocular injection with 0.3, 0.6, and 1.0 ml PFCL, respectively. All of the right eyes of the rabbits were in the control group. The morphological, electrophysiological and histological changes of the ocular tissue were observed 4, 8, and 12 weeks after the injection. Results No clinically significant retinopathy but only mild morphological changes were found in group 1 and 2, while obvious morphological and histological changes were found in group 3. Mild morphological and histological changes were found in all of the rabbits 4-8 weeks after the injection while significant ones were found 8-12 weeks after the injection. The results of electroretinography indicated a statistically significant decline of amplitude of b wave in group 3. Conclusions Long-term intraocular retention of few PFCL may cause mild histological changes but not affect the clinical function. Plentiful PFCL remains in eyes may lead to toxic reaction to the ocular tissue. (Chin J Ocul Fundus Dis, 2006, 22: 128-130)
Objective To summarize research progress of the mechanism of natural killer cells (NK cells) acted in regulating the T cell immunity in chronic infectious disease. Method Literatures about recent studies concerning how NK cells act as a regulator for T cells in chronic infectious disease were reviewed according to the results obtained from PubMed, Embase, CNKI, CBM, and Wanfang databases. Results NK cells that acted as regulators of T cell immunity could affect T cell immune responses through influencing antigen presentation, secreting cytokine, and presenting lytic activities, thus playing an important role in the immunological therapy of chronic infectious diseases. Conclusion NK cells are critical for T cell immune regulation, which could provide noval strategies for immunological therapy of chronic infectious disease, transplantation-related immune rejection, and autoimmune disease.