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Objective To detect the gene expressions of phosphatase and tensin homolog on chromosome ten (PTEN) engineered mesenchymal stem cells (MSCs) and observe their effects on DBTRG glioma cells.Methods Primary culture of human umbilical cord MSCs was performed;48 h after PTEN transfection into MSCs by liposomes,transfection results were detected under microscope.(1) Transfected MSCsPTEN were used as experimental group and MSCs as control group.PTEN protein and gene expressions of the cells from the two groups were detected by Western blotting and real time-PCR;soluble PTEN protein in the culture supematants was measured using ELISA.(2) In vitro cultured DBTRG cells were divided into four groups,and MSCs supernatant and 25％,50％ and 100％ MSCsPTEN supernatants were added into the four groups,respectively;24 h after culture,calcein AM/EthD-1 staining was used to detect the activity of DBTRG cells which co-cultured with different percentages of MSCsPTEN medium.(3) In vitro cultured DBTRG cells were divided into three groups,and MSCs supernatant and 25％ and 100％ MSCsPTEN supernatants were added into the three groups;RTCA was used to observe the growth curve of DBTRG cells.Results (1) A large number of red fluorescence masses were noted in the MSCsPTEN cells by microscope;real time-PCR and Western blotting indicated that the gene and protein expressions of PTEN in the experimental group were significantly higher than those in the control group (P＜0.05).PTEN content in the supernatants of the experimental group was significantly higher than that in the control group (P＜0.05).(2) CalceinAM/EthD-1 staining indicated that 25％,50％ and 100％ MSCsPTEN supernatant groups had significantly larger number of death DBTRG cells than MSCs supernatant group (P＜0.05);and with the increase of MSCsPTEN supeenatant percentages,the number of death DBTRG cells became larger (P＜0.05).(3) RTCA indicated that as compared with the MSCs supernatant group,25％ and 100％ MSCsPTEN supernatant groups had decreased DBTRG cell index.Conclusion After MSCsPTEN transfection,MSCsPTEN cells can stablely express targeted gene,and their supernatant can obviously promote the death of DBTRG cells and inhibit the growth of glioma cells;PTEN engineered MSCs may be an effective gene therapy for gliomas.
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Aim To explore the effects of curcumin on inflammatory reaction and blood-brain barrier permeability in rats following cerebral ischemic injury,and to further investigate its potential mechanisms.Methods SD rats underwent the cerebral ischemia/reperfusion injury by the sutrure occlusion model were randomly divided into sham-control,cerebral ischemia and curcumin-treatment groups.Neurological deficit scores,cerebral infarction volume,brain water content and blood-brain barrier (BBB) permeability were measured,myeloperoxidase(MPO)activities in rat brain were measured as an index of neutrophil infiltration;content of tumor necrosis factor-α(TNF-α)in rat brain was detected by ELISA;expression of matrix metalloproteinase-9(MMP-9) in rat brain was determined by Western blot.Results Neurological deficit and cerebral infarction volume was decreased in curcumin-treatment group.The degree of neutrophilicgranulo cyte infiltration in cerebral tissues was decreased and the integrity of BBB was improved.Curcumin could also inhibit TNF-α and MMP-9 expression.Conclusion Curcumin exerts the neuroprotective effect on cerebral ischemia/reperfusion injury in rats through inhibiting the inflammatory reaction and improving BBB integrity,which may be associated with the inhibiton of TNF-α and MMP-9.
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@#Objective To explore the microsurgical technique and clinical value of endoscope-assisted microsurgery via the transsphe-noidal approach to the clivus. Methods According to the results of microanatomy of endoscope-assisted via the transsphenoidal approach to sellar and clival area, the clinical data of 12 cases (8 with invasive pituitary adenoma, 3 with chordoma, 1 with chondroma) treated by transsphenoidal approach were studied retrospectively. All cases were followed-up 3 months to 6 years after operation. Results The tumor was totally removed in 8 patients, removed subtotally in 3 patients, and removed partially in 1 patient. 6 patients occurred transient diabetes insipidus, 2 patients with transient cerebrospinal rhinorrhoea. There were no death or intracranial infection. Postoperative follow-up was performed for 3 months to 6 years, No recurrence occurred except for enlargement of 1 chordoma. Conclusion Transsphenoidal approach satisfactorily and quickly reaches and helps remove the larger tumors in sellar and clival area without severe complication. It has the advantages of low incidence of surgical complication and high total removal rate. Endoscope-assisted may be helpful for this approach.