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Glioma remains the most common malignant tumor in the brain and is also the most difficult to treat. Immunotherapy achieving long-lasting tumor remission in multiple cancer types has received considerable attention due to its potential to improve the treatment outcomes of patients with glioma. However, clinical trials have not yet demonstrated major improvements in prognoses, which might be attributable to the extrinsic components and intrinsic mechanisms involved in the tumor microenvironment and immune system. It is particularly noteworthy that there is emerging evidence that current routine treatment modalities and the physical and psychological characteristics of patients have different impacts on the efficacy of glioma immunotherapy. This article addresses how these factors interact with the host immune system and tumor microenvironment, and highlights their potential roles in glioma immunotherapy, with the ultimate goal of developing better immunotherapybased personalized medicine strategies.
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Objective To investigate the cholinergic neuron properties and the changes of proliferation and apoptosis of PC 12 cell induced by different concentrations of all trans retinoic acid(RA). Methods PC 12 cell was induced by RA of concentrations of 1-50 ?mol/L. Changes of cell growth curve and cycle, apoptosis and changes of activities of acetylcholinesterase(AchE) and choline acyltransferase(ChAT) were detected by MTT assay, flow cytometry, TUNEL method, spectrophotometry and radiochemistry assay, respectively. Results ① RA significantly enhanced the activity of ChAT with concentrations between 1 to 20 ?mol/L. The activity of ChAT with concentration of 10 ?mol/L was higher than those with other concentrations. ② The apoptosis of RA induced PC 12 cells was markedly enhanced when concentration was higher than 10 ?mol/L. ③ A concentration dependent inhibition of proliferation was demonstrated in the RA induced PC 12 cells. ④ PC 12 cells proliferation was blocked in the G 1→S phase. Conclusion RA can be used to induce PC 12 cells to show the properties of cholinergic neuron with an optimal concentration of 10 ?mol/L.
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Objective To investigate the myelination of rat brain at different developmental stages.Methods Luxol fast blue staining,immunohistochemistry and Western blotting were applied to determine the distribution and alternation of myelin sheath in the brains of SD rats at different developmental stages.Results Negative LFB and MBP staining were shown at the stages of embryonicage 18 d(E18),postnatal age 0 d(P0) and P2.At P7,the corpus callosum was weakly stained by LFB.The positive stain of LFB and MBP occurred in P15 on the corpus callosum.With the development going on,the stain turned out stronger,especially in P30,which was similar with that in P90 and P720.The results of Western blot analysis showed the expression of MBP in rat brain was gradually increased with the development of rat brain.Conclusion The myelination starts before P15 in rat brain and becomes mature in P30.The optimal observation time of myelin is 30 d born after.
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Objective To investigate the effects of mutated PS-1 on the proliferation and apoptosis of RA-induced PC12 cells . Methods The MTT assay, flow cytometry, TUNEL method were used in the research. Results An inhibition of proliferation was demonstrated in the RA-induuced PC12 cells expressing PS-1 mutant L286V. The proliferation of the cells was blocked in the G1→S phase. Whether with or without fetal bovine serum(FBS), PS-1 mutant L286V was able to accelerate the apoptosis of RA-induced PC12 cells. This action was enhanced markedly under the culture without FBS. Conclusions The RA-induced PC12 cells expressing PS-1 mutant L286V shows that the proliferation is inhibited and blocked in the G1→S phase. Whether with or without FBS, PS-1 mutant L286V can accelerate the apoptosis of RA-induced PC12 cells. This action is enhanced markedly under the culture without FBS.
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Objective To study the relationship between GFAP and the malignant degree of astrocytoma. Methods After successful transfer of retrovirus with GFAP (PLBsKG) into C6 cells mediated with liposome, the morphology, growth curve and the proliferative cycle of the target cells were observed. The expression of the protein and mRNA of GFAP was determined with RT-PCR and immunocytochemical staining. In addition, the relationship between the staining GFAP-IR and the malignant degree of astrocytoma was analyzed according to Kermohan′s grading standard in 20 samples of the cells of astrocytoma with imaging analytic system. Results PLBskG resulted in the decrease of GFAP in the target C6 cells, and changed the morphology and increased the proliferation of C6 cells. The number of the cells in the G 2+M phages was increased. The intensity of GFAP-IR staining was negatively correlated to the malignant potency of astrocytoma. The more intensive staining of the cells, the lower of the malignant degree of astrocytoma and vice versa. Conclusion There is a close relationship between GFAP and malignant degree of astrocytoma. The level of GFAP expression serves the index of the malignant degree and prognosis of astrocytoma in certain degree.
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The changes of the morphology and percentage of the macroglia surrounding focus of brain stabbing injury (BSI) were observed by immunohistochemistry,immunoinfluorescence stain and flow cytometer(FCM) and the effect of magroglia during glial scar forming were elucidated.The results showed that a large number of GFAP-immunoreactive positive cells were accumulated around the focus of BSI.These cells were hyperplastic,hypertrophic,and emerged swollen cytoplasmic processes.The most marked changes were observed at 1-2 week after BSI.The results of FCM showed that the percentage of GFAP positive cells increased gradually and reached to a peak during 1～2 week after BSI.The peak ratio of GFAP positive was about 46%.However,the changes of morphology and number of GC positive cells were not detected after BSI.The authors believed that astrocyte is the main macroglia during glial scar formatting .The oligodendrocytes is not an active cell during this course.
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Objective To study the occurence, development and regulation of reactive gliosis with astrocyte (Ast) in vitro. Methods Ast was isolated and cultured in vitro and its model of reactive gliosis was established by scratching the cultured astrocytes. The reactivity and rules of Ast to injury was studied by morphological changes, RT-PCR, immunocytochemistry, in situ hybridization and imaging analysis. Results After scratching, the astrocytes showed typical features of reactive gliosis, with the hypertrophic cell body, thickened and lengtheded processes, and enhanced glial fibrillary acidic protein (GFAP) staining. In situ hybridization and RT-PCR analysis confirmed that the expression of GFAP mRNA was markedly increased. These changes occurred 1 d after scratching and reached the peak 5 to 7 d after injuring. Conclusion A model of reactive astrogliosis was successfully established in vitro which showed an active reaction to injury. The characteristics of reactive gliosis parallel that seen in vivo.
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Objective To investigate features and significance of the temporal and spatial expression of GFAP and Vimentin in olfactory system of Xenopus from metamorphosis to adult. Methods Xenopus tadpoles from stage 48 to 63 were made into serial sections(20??m)by Cryostat,each contains the nose,the olfactory nerve,and the olfactory bulb.The immunohistochemistry staining was done on these sections by anti-GFAP and anti-Vimentin,and then observed by fluorescence microscope.Results Olfactory nerve showed very strongly GFAP-IR staining during metamophosis of Xenopus.In the olfactory bulb,GFAP-IR positive staining was found only in the nerve layer,but not in glomeruli.By contrast,Vimentin-IR decorated radial glia in the olfactory bulb but faintly stained the olfactory nerve.Conclusion GFAP and Vimentin present complementary staining patterns,GFAP is expressed in the peripheral olfactory system while vimentin is expressed in the central part of olfactory system.