ABSTRACT
Objective To isolate the glioblastoma multiforme stem cells (GBM-SCs) from GBM specimens and to investigate the biological role ofmiR-153 in GBM-SCs so as to explore the application of gene therapy of GBMs.Methods CD133+ cells were separated using magnetic cell sorting technique (MACS) after primary culture.Immunofluorescence staining was employed to detect the CD133,nestin,glial fibrillary acidic protein (GFAP),microtubule-associated protein 2 (MAP2) expressions; real time-PCR was used to analyze miR-153 mRNA expression in CD 133-cells and CD 133+ cells.Lipofectamine RNAiMAX was used to transfect MiR-153 mimic (miR-153 group) and scrambled control oligonucleotides (NC group) into GBM-SCs; 7 d after that,sphere formation assay was performed to determine the self-renewal ability of GBM-SCs.Real time-PCR and immunofluorescence were carried out to examine the CD133,nestin,GFAP,MAP2 mRNA and protein expressions.At last,the proliferation ability of miR-153 treated GBM-SCs and NC cells was determined by CCK-8 at 24,48,72,96 and 120 h after the transfection and the apoptosis ratio was detected by flow cytometry 3 d after the transfection.Results GBM-SCs isolated from GBM specimens could express stem cell markers CD133 and nestin; after differentiation,the cells could express astrocyte marker GFAP and neuron marker MAP2; miR-153 expression in CD133+ cells was signficantly down-regulated as compared with that in CD133-cells (P＜0.05).Seven d after transfection,the number of spheres in the NC group was significantly larger than that in the miR-153 group (P＜0.05); real time-PCR indicated that the mRNA expressions of CD 133 and nestin in the miR-153 group were significantly decreased,and the GFAP and MAP2 mRNA expressions were statistically increased as compared with those in the NC group (P＜0.05).The results detected by immunofluorescence were in accordance with those by real time-PCR; 48,72,96h after the transfection,cell viability in cells from miR-153 group was statistically significant lower than that in the NC group (P＜0.05); flow cytometry showed that the apoptosis rate of cells from the miR-153 group (9.4 1％±1.98％) was significantlyhigher than that in the NC group (4.28％±0.31％,P＜0.05).Conclusion Reactivation of miR-153 expression suggests novel therapeutic strategy for GBM-SCs.
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Objective To study the factors affecting the prognosis of intracranial aneurysm operation for elderly patients. Methods Thirty elderly patients (≥60 years old) with intracranial aneurysm who were treated surgically were evaluated retrospectively. The factors that might affect the result of operation were analyzed by slng]e-factor and multi-factor. Result The factors that had significant correlation with the prognosis were Hunt-Hess grade,location of aneurysm,surgical timing (P =0.007,0.019,0.007).Conclusion The prognosis of intracranial aneurysm operation for elderly patients is affected by many factors,age is not a contraindication to aneurysm operation,early aneurysm operation treatment is suggested for elderly patients.
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BACKGROUND:At present,autogenous periosteum and artificial dura mater are usually applied as the substitute grafts for the dural defect by neurological surgery.However they do not accord with the developing trend of modern medicine,due to the limitations of material size and shape,operational complex and additional wound.OBJECTIVE:To observe and compare the evolution of a new type bio-artificial dura and autogenous periosteum in replacing orthotopic duraDESIGN:Controlled observation and trial.SETTING:Animal Testing Center in the 157 Hospital of Guangzhou City.MATERIALS:Nine New Zealand rabbits.aged 6 months and weighed 2-3 kg,either gender was selected.Twelve hybrid healthy dogs of both genders,aged 2 years and weighed 15-20 kg.New type dura mater(No.2006.3460627).METHODS:The experiment was carried out at the Animal Testing Center in the 157 Hospital of Guangzhou City from October 2003 to October 2005.After the general anesthesia and bilateral craniotomy,the bilateral dural defect and pia mater injury were induced partly,then dural neoplasty was performed using new type artificial dura and autogenous periosteum.MAIN OUTCOME MEASURES:At months 1,6,12 of modeling,each three rabbits were selected to isolate and expose the implanted materials,while each four dogs were selected at months 6,12,24 of modeling,died of disease or prior to death.General observation and microscopic assessment of samples were compared to analyze the development of implanted materials at difference stages.RESULTS:Except one experimental dog died during the anesthesia,9 rabbits and 11 dogs were involved in the final presented the extemal surface of adherence and separation with pedcranium,grew well with surrounding orthotopic dura.For the internal surface of materials,the new type artificial dura was more likely the orthotopic dura and did no adhere to pericranium, and filament-shaped adherence appeared occasionally, while there were filament-shaped even month 12 of grafting new type artificial dura into the experimental rabbits.inflammatory cellular reactions such as neutrophil and lymphocyte were not found,additionally no capsule wall formation occurred.The internal surface of artificial dura was covered with epithelial cells,which appeared fibroplasia,fibroblast proliferation,degradation of implants and obvious reduction of total cell amount.Moreover the blood capillary was also found.CONCLUSION:New type artificial dura can achieve the dural reconstruction through producing epithelial cells and being nibbled.degraded and substituted by autogenous tissue.And no adherence to cerebral tissues is found.New type artificial dura is superior to autogenous periosteum for repairing the dural defects.