Objective To observe the different effect such as high concentration of glucose and high concentration of insulin on GLUT1 of Rabbit Retinal Muuml;ller Cell in vitro. Methods Rabbit retinal Muuml;ller cells were cultured in vitro with suspended constitution，which were divided as the following groups: common control group，high glucose group，insulin group，high glucose combined insulin group. Laser confocal microscope combined with immunocytochemical and fluorescence staining method to quantitatively analyze the expression condition of GLUT1. Results The expression of GLUT1 has been enhanced obviously by high glucose and high insulin，which locates mainly in the cytoplasm that near to the nucleus. Conclusion Rabbit retinal Muuml;ller cells can express GLUT1，and the expression of GLUT1 can be reinforced by high glucose and high insulin. （Chin J Ocul Fundus Dis，2008，24：265-267）
Objective To investigate the effect of methylprednisolone on the expression of glial fibrillary acidic protein (GFAP) and proliferating cell nuclear antigen (PCNA) in Müller cells of rats’ retinae injured by laser. Methods Forty SD rats were randomly divided into two groups and inflicted with laser photocoagulation.The rats in treatment group were given methylprednisolone by intraperitoneal injection with a dose of 30 mg/kg for 3 days.At the 3rd,7th,14th,and 28th day after photocoagulation respectively, the eyes were enucleated,fixed and cut into sections.Immunohistochemical examination was used to detect the expression of PCNA and GFAP. Results After photocoagulation the Müller cells expressed PCNA both in the treatment and control group,and the expression of PCNA decreased sharply after 3 days. The expression of PCNA in treatment group was less than that in control group. After photocoagulation the Müller cells also expressed GFAP and the expression of GFAP lasted for at least 28 days ,and the expression of GFAP expression in the treatment group was less than that in the control group. Conclusion Methylprednisolone can reduce the expression of GFAP and PCNA in Müller cells of rats’ retinae injured by laser. (Chin J Ocul Fundus Dis, 2002, 18: 299-301)
Objective To observe the regulation effect of transforming growth factor alpha (TGFalpha;) on expression of glutamate transporter（GLAST）and ingestion activity of retinal Muuml;ller cells in mice. Methods To take the retinal tissue of Kunming mouse at postnatal 7～10 day, and then cultured Muuml;ller cells according to literature. The 3～4 generation cultured cells of the same primary cell were divided into two groups at random: ① TGFalpha; group: maintained in different concentrations of TGFalpha; as 50, 75, 125 and 150 ng/ml, 3 holes in each concentration;② Control group: cultured by Eagle culture medium which improved from Dulbeccon and contained 20% fetal calf serum. The influence of different concentrations TGFalpha; on GLAST activity in Muuml;ller cells were observed by L-3H-glutamate uptake detection; the expression of GLAST mRNA in Muuml;ller cells was determined by RT-PCR; the expression of GLAST protein was detected with immunocytochemical staining. Results With the increase of TGFalpha; concentration, both L3H glutamate uptake and GLAST mRNA expression were increased. The L-3H-glutamate accumulation had got to the maximum uptake at concentration of 125 ng/ml, which was 266% of that in control group, meanwhile, the expressions of GLAST mRNA also got to the maximum as 4 times of control group. Immunocytochemical staining indicated that the effect of 125ng/ml TGFalpha; on expression of GLAST protein was higher than that in the control group, the differences between two groups were statistically significant (Plt;0.05). Conclusion TGF-alpha; can increase GLAST activity through up-regulating the expression of GLAST mRNA and protein.
Objective To investigate the expression of induced heat shock protein (HSP) 70 in ratprime;s retinal neurons (RNs) and Muuml;ller cells, and evaluate the protective effect of HSP 70 on RNs injured with glucose deprivation and glutamate. Methods Ratprime;s RNs and Muuml;ller cells cultured in vitro were treated with heat shock (42℃ for 1 hour), and duration of the expression of HSP70 was detected by immunocytochemical techniques. Viability of the cells was measured by methyl thiazolyl tetrazolium (MTT) chromatometry after incitant toxic injury with glucose deprivation (0.56 mmol/L glucose for 6 hours) and glutamate (100 mu;mol/L for 6 hours). Simultaneously, the expression was interdicted by HSP70. Results Hypereffective expression of HSP70 was found in cultured RNs and Muuml;ller cells after heat shock. The viability of RNs pretreated by heat shock after injured with glucose deprivation and glutamate significantly increased which could be interdicted by HSP70 antibody. Conclusion Hypereffective expression of HSP 70 may be induced by heat shock, which enhances the ability of tolerance of RNs to the incitant toxic injury by glucose deprivation and exitotoxicity. (Chin J Ocul Fundus Dis, 2005,21:110-113)
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 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.
Objective To investigate the expression and clinical significance of T lymphocyte subsets, natural killer (NK) cells and CD19+ B cells in the elderly with primary immune thrombocytopenia (ITP) before and after treatment. Methods The elderly ITP patients diagnosed and treated in the Songjiang Hospital Affiliated to Shanghai Jiaotong University School of Medicine (preparatory stage) between January 2014 and June 2019 were retrospectively selected as the observation group. The healthy elderly in the same period were selected as the control group. According to the treatment, the observation group was divided into effective group and ineffective group. The expression levels of T lymphocyte subsets (CD3+, CD4+, CD8+ and CD4+/CD8+), NK cells and CD19+ B cells were observed and analyzed. Results A total of 75 subjects were included, including 35 in the observation group and 40 in the control group. The total effective rate was 85.71% (30/35). Before treatment, the expression levels of T lymphocyte subsets (CD3+, CD4+ and CD4+/CD8+) in the observation group were lower than those in the control group (P<0.05). There was no significant difference in other indexes between the two groups (P>0.05). After treatment, except for CD8+, the expression levels of T lymphocyte subsets (CD3+, CD4+ and CD4+/CD8+) in the observation group were higher than those before treatment (P<0.05). The expression levels of NK cells and CD19+ B cells were lower than those before treatment (P<0.05). The expression levels of T lymphocyte subsets (CD3+, CD4+ and CD4+/CD8+) in the effective group were higher than those before treatment (P<0.05), while the expression level of CD19+ B cells was lower than that before treatment (P<0.05). There was no significant difference in other indexes before and after treatment (P>0.05). There was no significant difference in the expression levels of T lymphocyte subsets (CD3+, CD4+, CD8+ and CD4+/CD8+), NK cells and CD19+ B cells in the ineffective group before and after treatment (P>0.05). Conclusions T lymphocyte subsets are abnormal in elderly ITP patients. The immune abnormality of T lymphocyte may be one of the reasons for elderly patients with ITP. With the improvement of therapeutic effect, immune cell subsets have also been improved.
Objective To develop a method for the primary culture of retinal Muuml;ller cells of adult rabbit in vitro. Methods Retina was isolated from adult rabbit, cut into 1 mm times; 1 mm pieces, and placed into Dulbecco modified Eagle medium/F12 containing 20% fetal bovine serum to culture. Cultured cells were identified by inverted phase contrast microscope, transmissim electron microscope and immunohistochemistry staining method. Results Visible cell processes grew out from the retinal tissues after three days culture, and more cells grew radically around the retina after seven days culture. The cultured cells were often inflated at one side and had one long process at another side, and the nuclei were elliptical and there were two or more than two nucleoli under inverted phase contrast microscope. The cytoplasm was rich and contained abundant microfilaments in eight to ten nanometers under transmission electron microscope. Immunohistochemistry assay showed that 95% of the cells were positive for glial fibrillary acidic protein and cellular retinaldehydebinding protein. Conclusion Rabbit retinal Muuml;ller cells can be cultured by the explant culture method.
Abstract: Objective To investigate the effect of cytokineinduced killer (CIK) cells immunotherapy on immunity function of non-small cell lung cancer (NSCLC) patients after operation. Methods Fifty patients with histological or cytological diagnosis of NSCLC on Ⅰstage,Ⅱstage andⅢa stage of tumor, nodes, metastasisclassification were randomly divided CIK cells therapy group and conventional therapy group, 25 cases each group. The immunity function of patients with NSCLC, including the levels of CD3+, CD4+ T cells, ratio of CD4+/CD8+, natural killer(NK) cells, and the levels of Th1/Th2 cytokine were detected before treatment, and the 2nd, 4th, 8th week after treatment. Results The levels of CD3+, CD4+ T cells, NK cells, ratio of CD4+/CD8+, interleukin-2(IL-2), interferon-γ(INF-γ) in CIK cells therapy group at the 2nd week after treatment were more higher than those before treatment (Plt;0.01), their levels reached the peak at 4th week, from then on, it began to decrease. Meanwhile, the levels of Th2 of CIK cells therapy group began to decrease at the 2nd week after treatment, a low ebb at 4th week. At the 2nd, 4th and 8th week,the levels of CD3+,CD4+ T cells, ratio of CD4+/CD8+, NK cells,IL-2, INF-γ, interleukin-4(IL-4), interleukin-10(IL-10) of CIK cells therapy group compared with those inconventional therapy group,there were statistical significance difference［(Plt;0.05),at 4th week after treatment, CD3+ 70.2%±9.1% vs.46.3%±5.8%; CD4+40.2%±7.1% vs.22.9%±4.5%; CD4+/CD8+ 1.82±0.43 vs. 1.09±0.34; NK 15.7%±5.4% vs.10.5%±2.5%; IL-2 34.8±11.7 ng/L vs. 19.8±12.1 ng/L; INF-γ63.7±23.3 ng/Lvs. 30.8±10.6 ng/L; IL-4 10.2±8.6 ng/L vs. 25.8±6.3 ng/L; IL-10 10.6±3.4 ng/L vs. 21.4±8.6 ng/L］. Conclusion The results indicate that CIK cells immunotherapy can enhance the immune function of NSCLC patients after operation.
Objective： To observe the effect of estrogen on the expr ession of pigment epithelium derived factor (PEDF) in cultured retinal Muuml;ller cells under the anoxic condition． Methods：After the anoxic retinal Muuml;ll er cells were tre ated with estrogen (E2) with the concentration of 10-6、10-5 and 10-7 mmol/L, t he level of expression of PEDF mRNA and the protein was detected by reverse tran scriptionpolymerase chain reaction and Western blotting analysis. Results：Th e expression of PEDF mRNA and protein decreased 24 hours after anoxia. E2 with t he concentration of 10-5 and 10-6 mmol/L inhibited the decrease of expression of PEDF mRNA and protein induced by anoxia, which related to the concentration of E2． Conclusion：strogen can regulate the expression of PEDF, which ma y play an important role in the regulation of retinal neovascularization．