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Objective:To study the expressione of autophagy-related protein Beclin-1 and LC3 in cortex and hippocampus of rats after cerebral ischemia reperfusion injury and the efficacy of Edaravone.Methods:Sprague Dawley rats were randomly divided into sham group,model group and Edaravone group.The cerebral ischemia reperfusion model was induced via Zea Longa with blocking the middle cerebral artery of 2 h and reperfusing of 24 h.Animals assigned to sham group were only separated left common carotid artery.Edaravone was injected intraperitoneally at a dose of 1 0 mg/kg at 15 min before reperfusion.The condition of nerve injury of rats was conducted by Neurobehavioral score.The degree of brain injury and success of model were determined based on 2,3,5-triphenyltetrazolium chloride(TTC)staining.The changes of neuron stained by hematoxylin and eosin (HE) in cortex and hippocampus were observed.The expression of Beclin-1 and LC3 was measured by immunohistochemistry.Results:After cerebral ischemia reperfusion the neurobehavioral score of edaravone group was(2.00± 0.67),which was obcviously less than(2.50± 0.53) of model group(P＜0.05).The infraction focus and the neuron injury in cortex and hippocampus neurons were also observed in model group,and the edaravone group reduced above expression.The positive rate of Beclin-1 of each group in cortex were (1 1.08± 0.85)％,(33.42± 1.57)％ and (25.61± 1.39)％,there was significant difference between model group with sham group and edaravone group (P＜0.05).The positive rate of Beclin-1 of each group in hippocampus were (10.34± 0.21)％,(31.82± 1.73)％ and (22.74± 1.26)％,there was significant difference between model group with sham group and edaravone group(P ＜ 0.05).The positive rate of LC3 of each group in cortex were (15.33± 0.47)％,(39.72± 1.73)％ and (28.53± 1.61)％,there was significant difference between model group with sham group and edaravone group(P＜0.05).The positive rate of LC3 of each group in hippocampus were (13.74± 0.37)％,(32.53± 1.43)％ and(25.38± 1.23)％,there was significant difference between model group with sham group and edaravone group (P＜0.05).Conclusion:The expression of Beclin-1 and LC3 was increased after cerebral ischemia reperfusion.Edaravone may reduce autophagy and brain injury through downregulation the expression of Beclin-1 and LC3.
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[ ABSTRACT] AIM:To investigate the changes of migration of lung adenocarcinoma cells promoted by IL-8 and the inner and outer mitochondrial membrane dynamic changes during this process.METHODS:Human lung adenocarci-noma cell line A549 was divided into control group and IL-8 group.Cell migration was analyzed by scratch detection and Transwell assay.The secretion of endogenous IL-8 was detected by ELISA.The protein levels of mitochondrial cytochrome C ( Cyt C) and mitochondrial outer membrane protein Tom20 was detected by Western blot.The mRNA expression of mito-chondrial fusion genes Mfn1, Mfn2 and OPA1 and fission genes Fis1, Drp1 and MTP18 was detected by RT-PCR.The morphological changes of mitochondria were observed by MitoTracker Red CMXRos dye staining and confocal microscopy. RESULTS:The migratory rate of A549 cells and endogenous secretion of IL-8 in A549 cells were higher than those in SPC-A-1 cells.The migratory rate of A549 cells was improved by IL-8 in a time-dependent manner.Compared with control group, the Tom20 protein expression was increased ( P0.05).Under confocal microscope, the punctate aggregates in the mitochondria of the A549 cells treated with IL-8 were observed.CONCLU-SION:The migratory rate of A549 cells is increased by IL-8, which is related to the changes of mitochondrial fusion genes and the fission genes.
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Visualized concept map can develop the ability of self-learning ability, knowledge integration ability and teaming and collaboration ability in medical students. Concept map is also a useful tool for improving student learning and clinical reasoning. Application of concept map in inte-grated problem based learning curriculum of basic medical sciences may promote meaningful learning and enhance the efficacy of PBL.
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Objective To investigate the role of human membrane associated sialidase (Neu3) in multidrug resistance ( MDR) of K562 cells. Methods K562 cells and K562/ADM cells were treated with DNR alone or with combination of DNR and NeuAC2en. The cell survival rate was measured by MTT assay; the expression of MDR related factors and apoptosis related factors were determined by Western blot and RT-PCR; Neu3 activity was detected by TBA reaction. Results The survival rate of K562/ADM cells was higher than that of K562 cells; NeuAC2en showed synergistic effect with DNR(P <0. 01) ; Neu3 activity of K562 and K562/ADM cells decreased after DNR or NeuAC2en induction and the decrease was most significant in the combination treatment group (P <0. 01). Under the identical condition, the protein expression of P-gp and Neu3 and the mRNA expression of MDR1, Neu3, BCL-xl and BCL-2 increased comparing with K562 cells. The mRNA level of BAX in K562 and K562/ADM groupsdid not changed significantly. MDR1, Neu3,BCL-xl and BCL-2 decreased after DNR induction and down-regulated most obviously in the groups treated by DNR combined with NeuAC2en. Conclusion Neu3 may be related with multidrug resistance of K562/ADM cell through regulating apoptosis and the expression of MDR1.
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Objective To observe the effects of Okinawa Habu apoxin protein-1 (OHAP-1) on the proliferation inhibition of rat C6 glioma cells and its mechanisms. Methods MTT colorimetric analysis was used to measure the inhibitory effect of OHAP-1 with different doses(2.5,5.0,and 10.0 mg?L-1) on C6 glioma cells .RT-PCR was used to evaluate the mRNA expressions of bcl-2 and bax genes.The activity of superoxide dismutase (SOD) and the level of maleicdialdehyde (MDA) in the C6 glioma cells were also examined. Results The proliferation of C6 glioma cells was significantly inhibited by different doses of OHAP-1(2.5,5.0,and 10.0 mg?L-1).The inhibitory rate were 49.77%,67.65%,and 76.42%,respectively.The inhibitory rate in 2.5,5.0, and 10.0 mg?L-1 groups were higher than that in control group(P
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AIM: To explore the molecular mechanism of brain tissue injury induced by lipopolysaccharide(LPS),the effects of panaxadiol(PDS) on the expression of nuclear factor kappa B(NF-?B) in cerebral cortex of rat with LPS shock were studied.METHODS: Rats were randomly divided into LPS group,LPS+dexamethasone group,LPS+PDS group and control group.The DNA binding activity and protein expression of NF-?B were observed.These indices were assayed at 1 h and 4 h after intravenous injection of LPS(4 mg?kg-1).RESULTS: EMSA showed that PDS inhibited NF-?B DNA-binding activity in nuclear extracts at both 1 h and 4 h after LPS injection,compared with the LPS group(P
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AIM:To study the effect of vitamin K3(VK3) on the induction of apoptosis in androgen-independent prostate cancer cell PC-3M in vitro.METHODS:Cell viability was estimated by MTT assay.AO/EB staining was performed to detect apoptotic cells.Apoptosis and the changes of cell cycle were detected by flow cytometry.NAC was used to observe the effect of growth inhibition by VK3.RT-PCR was used to confirm the changes in gene expression.Levels of intracellular peroxides were estimated by using an oxidation-sensitive fluorescent probe DCFH-DA.RESULTS:PC-3M cells growth was significantly inhibited by VK3(≥60 ?mol/L,P
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AIM: To investigate the expression of voltage-gated chloride channels (ClC)-3 protein and mRNA in human glioma specimen and its biological function. METHODS: The expression of C1C-3 was observed by immunohistochemical staining in 24 cases of human glioma, 4 cases of brain metastic cancer specimens and 5 cases of normal brain tissue as control; The C1C-3 mRNA expression were detected in the specimens with positive expression of ClC-3 protein by RT-PCR. RESULTS: ClC-3 protein was found negative in 4 cases of normal brain tissues and positive in 19 cases of human glioma and 4 cases of brain metastic cancer specimens. ClC-3 protein was mainly expressed in the membrane or cytoplasm of neoplastic cells and microvascular endothelial cells. The expression of ClC-3 mRNA was detected in 16 cases of human glioma and 4 cases of brain metastasis cancer specimens among the tissues with the positive expression of ClC-3 protein. The level of protein and RNA of ClC-3 in high malignant oligodendrogliomas was higher than that in low malignant ones. CONCLUSION: ClC-3 is generally expressed in human glioma and brain metastic cancer and is probably correlated with the classification of its pathological malignance.
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AIM: To explore the molecular mechanism of anti-shock effect of Shenmai (SM) injection and to observe its influence on TLR4 and I?B mRNA expression in hemorrhagic shock (HS) plus LPS rat lung. METHODS: Rats were divided into HS, HS+LPS, HS+LPS+ dexamethasone (DEX), HS+LPS+SM group, respectively. HS+LPS injury model of rats were induced by hemorrhagic shock (MAP 40 mmHg for 10 min) followed by intravenous administration of lipopolysaccharide (LPS, 1.5 mg/kg). Four hours later, the TNF-? contents in lung tissue were measured by ELISA, TLR4 mRNA and I?B? mRNA expression were assayed by RT-PCR, ultrastructural changes of the type Ⅱ alveolar cell were observed by electron microscope. RESULTS: The TLR4 mRNA expression level in HS+LPS+SM group was significantly decreased as compared to HS+LPS group (P