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AIM: To construct recombinant lentiviral vector with short hairpin RNA (shRNA) of CREB gene, and to investigate the effect of CREB gene silencing on mitochondrial morphology and cell apoptosis in oxygen-glucose deprivation/reoxygenation (OGD/R)-induced cortical neurons.METHODS: Three lentiviral vectors pLentiLox3.7 (PLL) inserted shRNA fragments targeting CREB gene were co-transfected with the packaging plasmids psPAX2 and pMD2.G to the 293T cells, and the virus particles, which was infected with the primary cortical neurons, was encapsulated.The protein expression of CREB was detected by Western blot.The mitochondrial morphology, cell apoptosis and the expression of Bcl-2 and Bax were evaluated by the methods of MitoTracker red, TUNEL and Western blot in OGD/R induced cortical neurons after CREB gene silencing.RESULTS: The pLL-CREB-shRNA1 was the most effective shRNA, which inhibited 80% CREB gene expression in the cortical neurons.The mitochondrial was appeared dot and fragment morphology in OGD/R induced cortical neurons with transfected pLL-CREB-shRNA1 plasmid.In addition, the expression of Bcl-2 was decreased, the expression of Bax, and the apoptosis of the neurons were increased by tranfected with pLL-CREB-shRNA1.CONCLUSION: CREB shRNA recombinant lentiviral vector specifically inhibits the expression of CREB gene.CREB gene silencing promotes the cell apoptosis and mitochondrial morphological changes in the cortical neurons induced by OGD/R.
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Objective Evaluating the effect of situational questions for qualifying the students'abilities in physiolo-gy examination .Methods Comparing the difficulty coefficients and discrimination indexes between situational and traditional choice questions .Results Compared with the traditional choice questions , the difficulty coefficient of situational questions increased , while the discrimination indexes were more reasonable .The discrimination indexes of situational understanding questions were higher than those of the traditional memory and understanding questions . There were no difference between discrimination indexes of the situational application questions and those of tradi -tional application questions .Conclusions Situational questions not only improved the quality of examinations , but also facilitate evaluating students'learning ability .
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Mitochondria are important intracellular energy supply organelles .As semi-autonomous organelles , the mitochondrial biogenesis , damage and clearance were the dynamic processes , which are dual-regulated by mitochondrial genes and nuclear genes , and maintain mitochondrial homeostasis according to the needs of the cells for energy .Recent studies provide evidence that the disorder of mitochondrial biogenesis in the neurons participates in the pathological process after cerebral ischemia/reperfusion, resulting in metabolic disturbance and cell apoptosis .This paper reviews the research progress of mitochondrion and cerebral ischemia/reperfusion injury .
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Objective: To investigate the effect of central interleukin-1? (IL-1?) on motor behavioral responses in novelty stress rats. Methods: The novelty stress was elicited by novel environmental stimuli with novelty stress box. The intracerebrolventricular (ICV) cannula and microinjection were performed with rat brain stereotaxic system. Movement behaviors of rats were monitored by behavioral radio-telemetry system. As behavior index, mean percent immobility (MPI) was used to assess immobility of rats. Results: The decrease of MPI was remarkably elicited by novel environmental stimuli. In non-stressful condition, ICV anti-IL-1? antibody did not influence the MPI. The decrease of MPI induced by novel environmental stimuli was significantly blocked by ICV pretreatment with anti-IL-1? antibody. In novel stress, MPI of rats was not affected by ICV injection of non-specific IgG. Conclusion: Central interleukin-1? plays an important role in modulation of motor behavioral response to novelty stress.
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Objective:To observe the thrombolytic effect of Naoxuebao in vitro and in vivo.Methods:The changes of thrombus in the carotid arterial,plasma fibrinogen (FIB),thrombin time (TT) and the diameter of blood clot were observed using Naoxuebao (three dose groups) after thrombosis.Results:With Naoxuebao,the time of blood clot was longer,the diameter of blood clot was shortened;carotid arterial thrombosis was dissolved.The contents of FIB decreased,TT was longer,and FIB and TT had negative correlation;whereas,PLG had no obious change.Conclusion:Naoxuebao had thrombolytic effect,but the effect had no relation with plasmin activating.
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AIM and METHODS:To study the effect of Nao-xue-bao at three different doses on blood coagulation，platelet aggregation by observing the changes in activated partial thromboplastin time (APTT), prothrombin time (PT), antithrombin-Ⅲ(AT-III), fibrinogen(Fng), plasminogen(Plg) and platelet aggregation(PAG). RESULTS:Compared with thrombosis group, the rats treated with Naoxuebao showed that the plasma APTT,PT were longer, and the activity of AT-III was increased. The content of Fng was reduced, TT was longer, there was a negative correlation between Fng and TT. Furthermore PAG-1, PAG-5 and PAG-M were reduced. CONCLUSION:Nao-xue-bao could inhibit thrombosis in different keys of blood coagulation.
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AIM: To observe morphological changes in transplanted intracerebral rat gliomas and rat survival time with gliomas under chemotherapy with angiotensin II-induced hypertension. METHODS: C6 glioma cells were cultured, and the effects of carmustine, nimodipine or/and teniposide on gliomas cells was observed. In addition, the brain tumor model was established in Wistar rats by stereotaxic inoculation of C6 glioma cells. The tumor-bearing rats were treated with carmustine, nimodipine lisplatin or/and teniposide during angiotensin II-induced hypertension, the pathological changes in gliomas was also examined. RESULTS: In vitro experiments showed chemotherapy resulted in morphologic changes in glioma cells, including cell enlargement, degeneration. In vivo experiments, the survival time of tumor-bearing rats was longer, the voume of gliomas was smaller in chemotherapy with hypertension group than those in chemotherapy alone, and pathological examination showed necrosis in the gliomas. CONCLUSION: Chemotherapy with angiotensin II-induced hypertension has a better inhibitory effect on rat intracerebral gliomas than chemotherapy alone.
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AIM: To investigate the activity of interleukin-1? converting enzyme in transplanted intracerebral rat gliomas under angiotensin II-induced hypertension chemotherapy. METHODS: The brain tumor model was produced in Wistar rats by stereotaxic inoculation of C6 glioma cells (1?10 12 /L). Tumor-bearing rats were treated with carmustine, teniposide and lisplatin (chemotherapy) during angiotensin II-induced hypertension. Then, the survival time of tumor-bearing rats, tumor blood flow, the concentration of drug, volume of gliomas and the activity of interleukin-1? converting enzyme in glioma were examined.RESULTS: The survival time of tumor-bearing rats was significantly longer in chemotherapy with angiotensin II-induced hypertension group than that of chemotherapy alone. In addition, regional tumor blood flow, the concentration of chemotherapeutic drug and the activity of interleukin-1? converting enzyme in transplanted rat gliomas were increased, while the volume of gliomas was decreased in hypertention chemotherapy group compared with chemotherapy alone. CONCLUSION: Chemotherapy with angiotensin II-induced hypertension has a enhancing effect on chemotherapy for improving the drug delivery to tumor tissue by a increased tumor blood flow and enhancing activity of interleukin-1? converting enzyme.
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AIM: To observe the role of free radicals in the inhibitory effect of chemotherapy on glioma cells. METHODS: C6 glioma cells were cultured in vitro , and treated with carmustine (B), teniposide (V), or/and nimodipine (N). Furthermore, the glioma-bearing rats were treated with B plus N, B+V+lisplatin (D)+N, or B+V+D+N+angiotensin Ⅱ. The MDA content and superoxide dismutase (SOD) activity in the culture supernatant and cortical brain tissue were assayed. RESULTS: B, V and N significantly decreased MDA content and SOD activity in the supernatant of glioma cell culture and C6 glioma cells. Chemotherapy reduced MDA content and increased SOD activity in the cortical brain tissue of tumor-bearing rats, with highest efficiency in B+V+D+N+angiotensin Ⅱ group. The survival time of tumor-bearing rats in B+V+D+N+angiotensin Ⅱ group was longer than that in other chemotherapy group. CONCLUSION: The antitumor effects of combined chemotherapy may be involved in the free radical metabolism. [
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AIM and METHODS: To study the effect of Nao-xue-bao at three different doses on blood coagulation,platelet aggregation by observing the changes in activated partial thromboplastin time (APTT), prothrombin time (PT), antithrombin-Ⅲ(AT-III), fibrinogen(Fng), plasminogen(Plg) and platelet aggregation(PAG). RESULTS: Compared with thrombosis group, the rats treated with Naoxuebao showed that the plasma APTT,PT were longer, and the activity of AT-III was increased. The content of Fng was reduced, TT was longer, there was a negative correlation between Fng and TT. Furthermore PAG-1, PAG-5 and PAG-M were reduced. CONCLUSION: Nao-xue-bao could inhibit thrombosis in different keys of blood coagulation.
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AIM: To study the central mechanism of intracerebral interleukin-1? in restraint stress-induced pressor response in rats. METHODS: Cardiovascular radio-telemetry system, stereotaxic microinjection system and neuroelectrophysiological methods were used to investigate the role of intracerebral interleukin-1? in pressor response induced by restraint stress, and the relation with the changes of discharge in the rostral ventrolateral medulla (RVL) neuron. RESULTS: The pressor response was induced by restraint stress and was reduced by intracerebral-ventricular injection of (icv) IL-1 receptor antagonist (IL-Ira) in conscious rats. The pressor response was directly induced by IL-1 (icv), which is related to increase of the extracellular discharge frequency in RVL neurons. CONCLUSION: Intracerebral IL-1? mediates pressor response induced by restraint stress, the mechanism may be closely related to RVL.
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AIM: To explore the role of calcineurin i n th e expression of NF-?B and the neurotoxicity in cultured cortical neurons treate d with interleukin-1? (IL-1?) and NMDA. METHODS: The cultured rat cortical neurons were used in the expe riment, damage of neurons was induced by interleukin-1?(IL-1?) or excitator y amino acid (NMDA). The degree of neuron damage was examined with the methods o f MTT assay and LDH releasing rate assay, as well as the Annexin V and PI immuno fluorescence. The expression of NF-?B p65 on the neurons was tested by the West ern blot analysis. RESULTS: Viability of neurons was obviously lower in the IL-1? group and NMDA group respectively than that in control group (P0.05). Annexin V and PI immunofluoresc ence showed that IL-1? mainly induced the neuron apoptosis, and NMDA induced th e neuron necrosis. CONCLUSIONS: The calcineurin mediates the higher expression of N F -?B p65 and neuron damage induced by IL-1?, but not play a critical role in th e necrosis induced by NMDA in the cultured cortical neurons. These results indic ate that calcineurin is the key molecule in the apoptotic signaling pathway.