ABSTRACT
AIM:TostudythechangeofradiosensitivityofU251cellsaftertreatedwithsodiumdichloroacetate ( DCA) and further to explore the possible mechanism .METHODS: The U251 cells were divided into 4 groups: control group, DCA group, X-ray irradiation without DCA pretreatment ( IR) group and X-ray irradiation with DCA pretreatment ( DIR) group.MTT assay was applied to determine the cell viability .The intracellular reactive oxygen species ( ROS) were detected by DHE fluorescence .The expression level of Bcl-2 was assessed by Western blot .The percentage of apoptosis of cells was determined by flow cytometry .RESULTS:No difference between control group and DCA group in cell viability (P>0.05) was observed.However, the cell viability of both IR group and DIR group was markedly reduced compared with control group ( P<0.05).Furthermore, the viability of DIR group was significantly decreased compared with IR group ( P<0.05 ) .The percentage of ROS positive cells was obviously increased in DIR group compared with IR group (P<0.05).The expression level of Bcl-2 was sharply decreased in DIR group (P<0.05) and the percentage of apoptosis of cells was significantly elevated ( P<0.05) in DIR group compared with IR group .CONCLUSION:The better antitu-mor effect was obtained by improving the radiosensitivity through pretreating the cells with DCA , and the possible mecha-nism was down-regulation of the Bcl-2 expression by developing the intracellular ROS .
ABSTRACT
Objective To comparatively study the differences of 18F‐FDG imaging agent by three kinds of different intravenous injection method for conducting PET / CT examination in aspects of the puncture success rate ,residual amount of drug injection and staff ray exposure time and their significance .Methods 240 patients with PET /CT examination were randomly divided into the group A ,B and C ,80 cases in each group .The drug injection adopted the traditional direct injection ,indwelling catheter injection and scalp venous needle connecting syringe(indwelling bubbles) .The puncture success rate ,drug residues and staff contacting radio‐pharmaceuticals time were compared among 3 groups .The obtained relevant data were statistically analyzed .Results The puncture success rate in the scalp venous needle connecting syringe (indwelling bubbles) and the indwelling catheter injection was higher than that in the traditional direct injection and the staff contacting radiopharmaceuticals time was significantly decreased ,the differ‐ences among them were statistically significant(P< 0 .01) ;the radioactive drugs residue in the scalp venous needle connecting syr‐inge was significantly decreased than that in other two kinds of injection method ,the difference was statistically significant (P <0 .01) .Conclusion The injection method of scalp intravenous needle connecting syringe (indwelling bubbles) significantly increases the puncture success rate ,reduces the radioactive drug residue ,at the same time decreases the staff radiation exposure time ,this method has the advantage in the radionuclide injection .
ABSTRACT
Cerebral vasospasm (CVS) is one of the serious complications of subarachnoid hemorrhage (SAH).Pathogenesis of CVS has not been fully clarified,and it may be associated with a variety of factors.With the development of molecular biology techniques,people have more understanding on SAH caused pathogenesis of CVS,and the research has also made considerable progress in the prevention of CVS.
ABSTRACT
Chronic ulcer of the lower extremities amounts for a grave and serious problem for public health. Western medicine focuses on controlling infection, improving blood circulation, surgical debridement, skin grafting, etc, but there are bottlenecks in the treatment. Traditional Chinese medicine (TCM) has a long history and a legacy of sound clinical efficacy in this area. TCM has developed a unique, effective external theory, and a large number of topical prescriptions and external technology. Through this research, a safe and effective treatment protocol of TCM for chronic ulcer of the lower extremities can be formed. To this end, during China's "Eleventh Five-Year" Plan, special research committees and projects on TCM external treatments and external technologies were established. This study on ulcer of the lower extremities constitutes one of the major research topics.
ABSTRACT
BACKGROUND: Recent researches indicate that ischemia and hypoxia can lead to abnormal brain metabolism and even energy failure, which is an important reason for brain damage and necrosis and identifies energy metabolism disorder as the key event in brain ischemia-reperfusion (IR)injury. Glucose transporter-3 plays the vital role in brain energy metabolism.OBJECTIVE: To observe the changes of cerebral infarct volume and glucose transporter-3 mRNA and protein expressions in cerebral cortical penumbra at different stages of focal cerebral ischemia and reperfusion in rats.DESIGN: Randomized controlled experiment.SETTING: Department of Neurosurgery, Second Hospital Affiliated to Sun Yat-sen University.MATERIALS: This experiment was conducted in the Animal Laboratory of Medical Research Center, Second Hospital Affiliated to Sun Yat-sen University between August and October 2002.Totally 56 SD rats were randomized into 3 groups which were subjected to ① ischemia for 1 hour followed by reperfusion (n=28), ② ischemia for 3 hours followed by reperfusion (n=24), and ③ sham operation (n=4). The rats in the first group were subdivided into 7 subgroups for examination at 1, 3, 6, 12, 24, and 72hours and 1 week after ischemia, with 7 rats in each subgroup; the rats in the second ischemia group were also subdivided in similar manner but without a 1 hour postischemic subgroup. The rats in the sham operation group only received the operation but without arterial occlusion.METHODS: Focal cerebral ischemia-reperfusion (IR) injury model was induced in the rats in the two ischemic groups by means of insertion of suture for arterial occlusion, and the ratio of central ischemic area to cerebral infarct volume in the ischemic penumbra was examined at the specified time points. Reverse transcription-PCR (RT-PCR) was used to detect the expression of glucose transporter-3 mRNA in the cerebral cortex in ischemic penumbra region, and semi-quantitative immunohistochemistry (IHC) employed to detect the level of glucose transporter-3 protein.MAIN OUTCOME MEASURES: Cerebral infarct volume after IR injury, changes of transporter-3 mRNA and protein expressions after IR injury.RESULTS: Totally 56 rats were used in this experiment and all entered result analysis. The post-IR cerebral infarct volume was obviously smaller in 1-hour ischemia group than in 3-hour ischemia group. Glucose transporter-3 mRNA expression began to increase 3 hours after ischemia in 1-hour ischemia group, reaching the peak level at 24 hours and still mainrained higher level than that of the sham operation 1 week; in 3-hour ischemia group, the mRNA expression was slightly decreased at 3 hours but began to increase afterwards till reaching the peak level at 24 hours, followed then by recovery of normal level at 1 week. The changes in glucose transporter-3 protein and mRNA expressions followed almost the same pattern.CONCLUSION: Glucose transporter-3 expression is up-regulated in the ischemic penumbra region, possibly as a protective response to cerebral IR injury.
ABSTRACT
AIM:To study the effects of neurological improvement and remyelination after intracranial hemorrhage(ICH)in rats by a novel therapeutic strategy with hepatocyte growth factor(HGF)gene transfected human umbilical cord mesenchymal stem cells(hUCMSCs)by lentiviral vector.METHODS:ICH was induced in 60 adult male Sprague-Dawley rats by a stereotactically guided injection of bacterial type IV collagenase into the right internal capsule.Non-modified hUCMSCs,HGF-modified hUCMSCs with lentiviral vector or PBS were administered left intraventricularly 7 d after right internal capsule ICH.All rats underwent modified neurological severity scores for 35 d.Luxol fast blue staining,immunohistological staining and Western blotting assessments for myelin basic protein(MBP)were applied.RESULTS:The ICH rats receiving HGF-modified hUCMSCs demonstrated significant functional recovery,determined by modified neurological severity scores,compared to the other groups from 2 weeks after cell therapy.As indicated by Luxol fast blue staining,the percent area of demyelination was obviously reduced in the HGF-hUCMSC treatment group compared to the PBS control group and hUCMSC-only treatment group at 5 weeks after ICH.The expression of MBP detected by immunohistological staining and Western blotting was significantly higher in HGF-hUCMSCs treated brain than that in other groups(P
ABSTRACT
AIM: To investigate the volume percentage of infarct and expression level of glucose transporter-3 (GLUT3) transcription and protein at different ischemic time points and different reperfusion time points in rat focal cerebral ischemic penumbra. METHODS: Focal ischemic models of middle cerebral artery occlusion (MCAO) in rats were made by inserting nylon thread. Brain samples were harvested from ischemic penumbra. Infarct volume was analyzed quantitatively by Kontron IBAS 2.5 image auto-analyses system. The change of GLUT3 mRNA was assessed by RT-PCR, and the expression of GLUT3 protein was assessed by immunohistochemistry. RESULTS: The infarction volume in MCAO 1 h/R group was obviously smaller than that in MCAO 3 h/R group. GLUT3 began to ascend at 3 h in MCAO 1 h/R group, reached to climax at 24 h and remained higher than normal at 1 week. In contrast, in the MCAO 3 h/R group, GLUT3 had a descent at 3 h. Later on, it ascended rapidly, and reached climax at 24 h. At 1 week, it approached to normal. The expression level of GLUT3 protein corresponds with that of mRNA. CONCLUSION: GLUT3 expression is up-regulated in the penumbra region after focal cerebral ischemia, it may be a protective reaction against ischemia/reperfusion injury. [
ABSTRACT
AIM: To investigate the volume percentage of infarct and expression of glucose transporter-1 (GLUT1) transcription and protein levels at different ischemic time point and different reperfusion time point in rat focal cerebral ischemic penumbra. METHODS: Focal ischemic models of middle cerebral artery occlusion (MCAO) in rats were made by inserting nylon thread. Brain samples were harvested from ischemic penumbra. Infarct volume were analyzed quantitively by Kontron IBAS 2.5 image auto-analyses system. The expressien of GLUT1 mRNA was assessed by RT-PCR, and the expression of GLUT1 protein was assessed by immunohistochemistry. RESULTS: The infarction volume in MCAO 1 h/reperfusion (R) group was obviously smaller than that in MCAO 3 h/R group. GLUT1 increased at (1 h) MCAO 1 h/R group, climbed to climax at 24 h and remained higher than normal at 1 week. In contrast, in the MCAO 3 h/R group, the corresponding index was at 3 h, 24 h and 1 week, but the increasing degree of GLUT1 was slighter than MCAO 1 h/R. GLUT1 protein began to ascend at 1 h, reached climax at 24 h and was higher than normal at 1 week in MCAO 1 h/R group, while in MCAO 3 h/R group, the corresponding index was at 3 h, 24 h and 1 week. CONCLUSION: GLUT1 expression is notably up-regulated in the penumbra region after focal cerebral ischemia, it may be a protective reaction against ischemic injury. [
ABSTRACT
Objective:After construction and identification of HBsAg gene recombinant backbone adenoviral vector,it is to prouduct HBsAg gene recombinant adenoviruses by packing PAd-Easy-1-HBs in 293 cells.Methods:The gene of interest was amplified from plasmid pEcob-6 PCR, the gene of interest which contained HBsAg gene was cloned into on adenoviral shuttle vector pAd-track-cmv. The pAd-track-cmv-HBs was linearized by digesting with restriction endonuclease Pme-1, and subsequently cotransformed into E.coli BJ-5183 cells with an adenoviral backbone vector pAd-Easy-1, Homologous recombinants were performed in bacterial cells. Finally, the linearized backbone adenoviral vector was transfected into adenoviruses packing cells lines,e.g. 293 cell by lipofectamine transfection. Transfections and viral productions can be minotored by green fluorescent protein(GFP). The expression of HBsAg in supermatant was investigated by ELISA. It was a certain HBsAg vaccine to amplify recombinant adenoviruses by repeating the infevtion cell to collect the viral supermatant.Results:GFP expression was visible by fluorescence microscopy after transfection. Adenoviral titer was monitored by GFP expression. GFP expression was visible after repeating the infection cell using the viral supernatant in more than 90 percent of the cells.The HBsAg also expressed in supermatant.Conclusion:HBsAg gene recombinant adenoviral backbone vector has been constructed successfully. HBsAg gene recombinant adenoviruses have been producted by packing in 293 cells. The study provides the possibility of further researches on the development of new anti-HBV vaccines.