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.
Neural stem cell is a kind of stem cells that can differentiate into neural and glial cells. While Müller cells, the main endogenous neural stem cell in retina,have the features to reentry into the cell cycle and differentiate into neural cells after retinal damage. Although it is highly effective for retinal Müller cell differentiation spontaneously after retinal injury in vertebrates, this feature is rigorous restricted in mammals. Recently, some transcription factors,such as Ascl1, Sox2, Lin28, Atoh7, are sufficient to drive quiescent Müller cells back in proliferation to generate new retinal neurons. Moreover, combining Ascl1 expression with a histone deacetylase inhibitor can bypass the limitation and increase the generation of new neurons in the adult retina. These regenerated neurons integrate the existing neuronal network and are able to respond to light, indicating that they can likely be used to restore vision. While these results are extremely promising, the regenerative response is still limited, likely because the proliferative capacity of mammalian Müller cells is low compared to their zebrafish counterparts. It is indeed necessary to identify new factors increasing the efficiency of the regenerative response.
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.
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)
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 investigate the safety and clinical efficacy of dendritic cell (DC)-cytokine induced killer (CIK) cell adoptive immunotherapy combined with chemotherapy in patients with gastric cancer after radical gastrectomy.MethodsForty-eight patients with gastric cancer after the radical gastrectomy receiving the DC-CIK cell adoptive immunotherapy combined with XELOX or FOLFOX chemotherapy were enrolled as a study group in the First Hospital of Lanzhou University from January 2014 to January 2016. In addition, 48 patients with gastric cancer after the radical gastrectomy in the same period and only receiving XELOX or FOLFOX chemotherapy were collected as a control group. The CD3+, CD3+CD4+, CD3+CD8+, CD3–CD56+ (NK cell), and CD3+CD56+ (NKT cell), toxic reaction, quality of life were evaluated in both groups before and after the treatment, and the long term effect were compared in both groups.Results① There were no significant differences in the gender, age, clinical stage, etc. between the two groups (P>0.05). ② The CD3+, CD3+CD4+, CD3+CD8+, CD3–CD56+, and CD3+CD56+ cells in the peripheral blood had no significant changes between before and after treatment in the study group (P>0.05), which were decreased after the treatment in the control group as compared with before the treatment and were significantly lower than those in the study group (P<0.05). ③ The levels of CEA, CA19-9, and CA724 in the peripheral blood after the treatment in the study group and the control group were significantly lower than those before the treatment (P<0.05), which in the study group were significantly lower than those in the control group after the treatment (P<0.05). ④ The incidences of leukopenia, thrombocytopenia, and diarrhea in the study group were significantly lower than those in the control group (P<0.05). ⑤ Compared with before the treatment, the body function and emotional function after the treatment were significantly improved in the study group (P<0.05). And in the body function, emotion function, role function, cognitive function, and social function were significantly improved than those in the control group (P<0.05) after the treatment. ⑥ The progression-free survival in the study group was significantly better than that in the control group (P<0.05). There was no significant difference in the overall survival between the study group and the control group (P>0.05).ConclusionDC-CIK cell adoptive immunotherapy combined with chemotherapy could significantly improve immune status and quality of life of patients with gastric cancer after radical gastrectomy, reduce adverse effects of chemotherapy, improve long term effect, and prolong progression-free survival.
【Abstract】Objective The effects of tumor infiltrating lymphocytes (TIL) on cellular immunologic function of patients with breast cancer were studied. Methods Twenty five patients with breast cancer were treated by the TIL that were isolated from tissue of tumor. T cell subgroups and natural killer cell (NK cell) activity of peripheral blood, the levels of serum soluble interleukin-2 receptor (sIL-2R) were assayed before and after treatment. Results CD3, CD4, CD4/CD8 and NK cell activity were ascended obviously, and CD8, sIL-2R were descended obviously after the treatment of TIL. Conclusion TIL can enhance the cellular immunologic function of patients with breast cancer.
ObjectiveTo investigate the influence of programmed cell death protein-1 (PD-1) monoclonal antibody on the anti-lung cancer effect of cytokine-induced killer cells (CIK) which were programmed in vitro. MethodsPeripheral blood mononuclear cells from 20 patients (8 males and 12 females with an average age of 56.45±5.89 years ranging from 42 to 65 years) diagnosed with advanced lung cancer from January to May 2019 at the Department of Oncology of Dalian Central Hospital were collected and induced to amplify into CIK cells in vitro. PD-1 monoclonal antibody combined with CIK cell culture group, individual cell culture group and PD-1 monoclonal antibody group were set up to detect the cell killing activity of CIK cells against lung cancer under different effective target ratio conditions, and the ratio of perforin and granzyme positive expression in PD-1 monoclonal antibody combined CIK cell culture group and individual CIK cell culture group was detected by flow cytometry. ELISA method was used to detect the interleukin-2 (IL-2), interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) cytokine secretion levels in the two groups.ResultsThe killing effect of CIK cells on A549 lung cancer cells increased with the increase of effective target ratio by CCK8, and PD-1 monoclonal antibody increased the killing effect of CIK cells on A549 lung cancer cells under different effective target ratio, E∶T=5∶1 (28.5%±1.9% vs. 20.3%±1.8%), 10∶1 (40.6%±2.4% vs. 31.7%±2.1%), 20∶1 (57.4%±3.5% vs. 44.7%±3.8%), 40∶1 (74.1%±8.3% vs. 60.8%±5.3%). The killing effect of PD-1 monoclonal antibody combined with CIK cells and CIK cells alone on A549 lung cancer cells was statistically different (P<0.05). The killing effect of cells in both groups on lung cancer A549 cells was stronger than that of the PD-1 monoclonal antibody group (P<0.01). The results of flow cytometry showed that PD-1 monoclonal antibody increased the positive ratio of perforin and granzyme release in CIK cells, and the positive ratios of perforin release (46.7%±3.5%% vs. 35.1%±2.2%) and granzyme release (34.6%±3.8% vs. 25.7%±3.3%) in PD-1 monoclonal antibody combination with CIK cells group and CIK cells group were statistically different (P<0.05). Similarly, the secretion levels of IL-2, TNF-α, and IFN-γ cytokines were also increased in the PD-1 monoclonal antibody combined with CIK cells group compared with the CIK group (5 409.0±168.8 pg/mL vs. 4 300.0±132.3 pg/mL, 252.7±16.7 pg/mL vs. 172.5±8.6 pg/mL, 327.2±23.5 pg/mL vs. 209.7±16.0 pg/mL, P<0.05).ConclusionPD-1 monoclonal antibody can promote the release of tumoricidal substances in CIK cells and improve the killing effect of CIK cells on lung cancer A549 cells. It is speculated that the infusion of PD-1 monoclonal antibody before CIK cell adoption in lung cancer patients may be more beneficial to the treatment of disease. PD-1 monoclonal antibody combined with CIK cell therapy is promising as a new type of lung cancer immunotherapy.
Objective To observe the influence of interleukin-1beta; (IL-1beta;) on the expression of phosphorylated signal transducers and activators of transcription 3 (pSTAT 3) in rat retinal Muuml;ller cells.Methods For in vitro study cultured Muuml;ller cells were treated with IL-1beta; of different concentrations (0, 0.1, 1, 5 and 10 ng/ml) for 24 hours. For in vivo study, 32 Sprague-Dawley（SD）rats were divided into 4 groups randomly (control group，100，500 and 1000 ng/ml group) with 8 rats in each group. After 24 hours of injection with phosphate buffered solution (PBS), or 100,500,1000 ng/ml IL-1beta; into the vitreous cavities of the above rats, retinas were harvested. The expressions of pSTAT3 in cultured Muuml;ller cells or treated retinas were evaluated by indirect immunofluorescence and western blotting.Results After 24 hours of incubation without IL-1beta;, pSTAT3 has little expression in cultured Muuml;ller cells, but was upregulated by 1 ng/ml or higher IL-1beta; in a dosagedependent manner (F=46.64, 43.78;P＜0.01). pSTAT3 was not expressed in adult rat retina, but was upregulated by vitreous injection of 100 ng/ml or higher IL-1beta; in a dosagedependent manner (F=73.53,43.70;P＜0.01).pSTAT3 expressed mainly in inner nuclear layer and ganglion cell layer. Doublelabeling showed that there was no costaining of pSTAT3 and glial fibrillary acidic protein (GFAP) in retina of control group, but there were many costained Muuml;ller cells in retinas treated with IL-1beta;.Conclusions Expression of pSTAT3 in Muuml;ller cells could be activated by IL-1beta; which may represent one pathway link to reactive gliosis.
Obiective
lt;brgt;To investigate the change of the activity of proliferation in cultivated Muuml;ller cells treated by advanced glycation endoproducts (AGEs), and the effect of these changes on expression of occludin in bovine retinal vascular endothelial cells (BREC).
lt;brgt;Methods
lt;brgt;The cultivated Muuml;ller cells were devided into normal growth group and cultured with AGEs group. The cultured BREC were devided into 4 groups:group 1, without any medium; group 2, with normal growth Muuml;ller cell medium (MCM); group 3,MCM treated by AGEs; group 4, without cell as the control. Enzyme-linked immuno sorbent assay was used to detect the content of occludin in the medium in the 4 groups.
lt;brgt;Results
lt;brgt;The content of expression of occludin was the most in group 2, less in group 1, and the least in group 3.
lt;brgt;Conclusion
lt;brgt;AGEs may promote the abnormal proliferation of Muuml;ller cells and inhibit the expression of occludin in BREC.
lt;brgt;(Chin J Ocul Fundus Dis, 2006, 22: 28-30)
