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Objective To investigate the characteristics of contingent negative variation(CNV) in patients with subcortical ischemic vascular cognitive impairment (SIVCI) and their relationship with continuous performance task (CPT) to establish a neuroelectrophysiological index of attention impairment in earlier period of Vascular Cognitive Impair-ment. Methods CNV was assessed in 45 SIVCI patients [30 patients with subcortical ischemic vascular cognitive impair-ment no dementia (SIVCIND ) and 15 patients with subcortical ischemic vascular dementia (SIVD )] and 15 normal con-trol (NC) by using a EB-Neuro ERP Instrument. Results ①CNV in SIVCI group were less regular compared with nor-mal control;CNV in SIVCIND group had decreased Expectancy Wave (EW) amplitudes (9.98± 4.10μV vs. 16.13±2.75μV) and EW areas (14848.10 ± 3199.16 μV · ms vs 20058.87 ± 1025.95 μV · ms) compared with normal control (P0.05);②Pearson correlation\Spearman rank correlation analysis revealed that there were significant, positive correlations between EW latency and CPT Reacting time(R=0.748, P0.05)、EW amplitude(R=-0.191,P>0.05)and there were significant, negative correlations between EW area and CPT Reacting time(R=-0.718,P<0.01)、CPT missing rate(R=-0.829 ,P<0.01), EW amplitude and CPT Reacting time(R=-0.616, P<0.01). Conclusion SIVCI patients in early stage have attention deficits in sustained attention and CNV EW ar-ea may be a good neuroelectrophysiological index for sustained attention impairment .
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Nuclear transfer (NT) is a new cloning technology developed in recent years. NT methods consist of electrofusion, NT mediated by polyethylene glycol (PEG) and microinjection. The success of somatic nuclear transfer depends on the source of donor nucleus, developmental stage of recipient cytoplasts, cell cycle synchrony of donor nucleus. Different methods of harvesting cells have effect on the efficiency of NT. The somatic nucleus will be reprogrammed after NT and will restore a totipotent state in order to undergo development.
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Animais , Humanos , Diferenciação Celular , Fisiologia , Divisão Celular , Células Cultivadas , Reprogramação Celular , Clonagem de Organismos , Transferência Embrionária , Microinjeções , Técnicas de Transferência Nuclear , Oócitos , Biologia Celular , FisiologiaRESUMO
Objective:To explore the diagnostic value of brainstem auditory evoked potential(BAEP) in patients with vertebrobasilar transient ischemic attack (TIA). Methods: BAEP was performed in 33 patients with vertebrobasilar TIA. Results: The abnormality rate of BAEP was 75.8%. It showed that theⅠ,Ⅲ or Ⅴ wave was abnormal in some patients. The peak latency (PL) of Ⅰ,Ⅴ waves and the interpeak latency (IPL) of Ⅲ～Ⅴ and Ⅰ～Ⅴ wave from vertebrobasilar TIA was longer than the control group (P
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BACKGROUND: Transplanted bone marrow mesenchymal stem cells (BMSCs) migrate to the injured regions and exert their therapeutic effects. The specific mechanisms involved in their directional migration to lesions remain unclear. OBJECTIVE: To investigate the chemokine receptor CXCR4 and CX3CR1 expression of human BMSCs in hypoxia culture. DESIGN, TIME AND SETTING: The cytology in vitro study was performed at the Central Laboratory, Xingqiao Hospital, Third Military Medical University of Chinese PLA from February 2008 to February 2009. MATERIALS: Cells harvested from the iliac heparinized bone marrow were obtained by iliac crest aspiration from healthy adult volunteers, aged 15 to 40 years old, at the Department of Hematology, Xinqiao Hospital, Third Military Medical University of Chinese PLA. METHODS: Bone marrow was obtained by puncture. Human BMSCs were harvested by combination of density and gradient centrifugation and different adherent method. Cells at passage 3 were incubated in a 25 cm2 flask. When 70%-80% confluence was found, cells were incubated at 37 ℃ and saturated humidity in an incubator containing 3% O2, 5% CO2, 92% N2 for 48 hours, and those incubated under normal oxygen as controls. MAIN OUTCOME MEASURES: Morphology was examined by phase contrast microscopy. Cell surface markers were tested by flow cytometer. The CXCR4 and CX3CR1 mRNA expression were detected by real-time PCR. The CXCR4 and CX3CR1 protein expression were determined by Western blot assay. RESULTS: All of the cells had a fibroblast-like morphology cultured in vitro and reached 90% confluence at 12-14 days, with the presence of polarity arrangement and whirlpool-shape. Cells were uniformly positive for CD105 (99.38%) and CD29 (99.13%), but negative for CD14 and CD45. Exposure of BMSCs to 3% O2 increased expression of the CXCR4 mRNA and CX3CR1 mRNA, which were respectively 2.130 times and 2.361 times of normal culture; expression of the CXCR4 protein and CX3CR1 protein was respectively 1.69 times and 1.93 times of normal culture. CXCR4 and CX3CR1 mainly expressed in membrane and cytoplasm of human BMSCs. CONCLUSION: Hypoxia (3% O2) can upregulate the expression of CXCR4 and CX3CR1 in human BMSCs, which might be one of the machenisms underlying the migration of BMSCs.
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BACKGROUND:Recent research has shown that transplanted bone marrow mesenchymal stem cells(BMSCs) migrate to the injured regions and exert their therapeutic effects in cases of intracranial trauma, stroke, inflammation and degenerative disease.The specific mechanisms involved in their migration to lesions are still to be fully elucidated.OBJECTIVE:To explore the effects of stromal cells derived factor-1(SDF-1) and its receptor CXCR4 on the migration of transplanted BMSCs to ischemic brain lesions.DESIGN, TIME AND SETTING:The cytological in vivo study was performed at the Central Laboratory, Xinqiao Hospital, Third Military Medical University of Chinese PLA from February 2008 to February 2009.MATERIALS:Bone marrow samples were obtained from normal or primary affection non-involved bone marrow patients aged 15-40 years at the Department of Hematology, Xinqiao Hospital, Third Military Medical University of Chinese PLA.A total of 72 healthy male Sprague Dawley rats aged 3-4 months were supplied by the Experimental Animal Center, Research Institute of Surgery, Third Military Medical University of Chinese PLA.METHODS:Human BMSCs were isolated by combination of gradient centrifugation and different adherent time method.The transient middle cerebral artery occlusion(MCAO) was induced using intraluminal vascular occlusion in 54 rats, based on the method described by Nagasawa et al.The remaining 18 rats served as sham operation group, only inserted with thread for 10 mm depth.At 2, 4 and 8 days after cerebral ischemia, the expression of SDF-1 in the ischemic brain was determined by real time RT-PCR and immunohistochemistry in 9 rats from either group.The remaining 36 rat models of cerebral ischemia/reperfusion were equally and randomly assigned into a cell transplantation and solution control groups.1 mL human BMSCs(2?109/L cells) or 1 mL phosphate buffered saline were slowly infused through the caudal vein at 24 hours following reperfusion.MAIN OUTCOME MEASURES:CXCR4 mRNA and protein expression in human BMSCs was determined.SDF-1 mRNA and protein expression following ischemia/reperfusion were detected.Migration of transplanted human BMSCs into the damaged region was observed through immunohistochemistry.RESULTS:RT-PCR showed that human BMSCs were positive for CXCR4 mRNA.Immunocytochemistry revealed that CXCR4 mainly expressed in cell membrane and cytoplasm of human BMSCs.At 2, 4 and 8 days following cerebral ischemia/reperfusion, SDF-1 mRNA levels showed an increased tendency, and showed significant difference compared with the sham operation group(P
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Objective To investigate the distribution and migration of bone mesenchymal stem cells (BMSCs) in the brain after intravenous injection into brain-ischemic rats and to explore the effects on the learning and memory in rats. Methods Bone mesenchymal stem cells from adult rats were cultured in vitro. Then the cells were labeled by Hoechst 33342 and infused into the adult rats on day 3 after the ischemia/reperfusion model was established by cerebral artery occlusion. Morris water maze test was used to explore the effects on learning and memory function of the rats at 3 weeks after transplantation. Fluorescence microscope was used to identify BMSCs in rat brains at 4 weeks after transplantation. Results All indices in Morris water maze test were significantly different from those of control group at 3 weeks BMSCs transplantation(P
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Objective To investigate the induction mechanism of MT ⅢmRNA in the brain after cerebral ischemia.Methods The forebrain ischemia reperfusion model was established in rats. The changes of the expression of MT Ⅲ mRNA in hippocampus after forebrain ischemia reperfusion were observed by in situ hybridization method. The changes of free Zn 2+ in hippocampus after forebrain ischemia reperfusion was examined using Zn 2+ specific fluorescent probe(TSQ). The Zn 2+ chelator (CaEDTA) was injected into the lateral ventricles for determining influences of Zn 2+ on the MT Ⅲ mRNA expression and the neuronal damage after forebrain ischemia/reperfusion.Results (1) The expression of MT ⅢmRNA in hippocampus increased gradually after cerebral ischemia and reached the peak in 96 hours after reperfusion. Seven days after reperfusion the expression of MT ⅢmRNA reduced to the normal level. (2) Zn 2+ fluorescence in the hilus of dentate gyrus, CA 3 region and the stratum radiatum and stratum oriens of CA 1 decreased slightly at 48 hours after reperfusion. From 72 to 96 hours after reperfusion, the fluorescence returned to normal, but some new fluorescence dots appeared in pyramidal neurons of CA 1 and the hilus of dentate gyrus increased gradually. Seven days after reperfusion, the fluorescence returned to normal. (3) The cell membrane impermeable Zn 2+ chelator could reduce the intracellular concentration of free Zn 2+ and the expression of MT Ⅲ mRNA.Conclusion The expression of MT Ⅲ mRNA can be induced by the increase in the concentration of intracellular free Zn 2+ after forebrain ischemia/reperfusion.
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Objective To explore the relationship between the serum neuron-specific enolase(NSE) level and stroke region and size,the degree of neurological deficit and prognosis at acute stage of stroke.Methods The level of serum NSE was detected by ELISA in 96 stroke patients at acute stage,including 58 cases of cerebral infarction (CI),27 cases of intracerebral hemorrhage(ICH),11 cases of subarachnoid hemorrhage (SAH) and 25 normal controls(NC group).The relationship between the level of serum NSE and China Stroke Score (CSS),Barthel index(BI),score of Glasgow-Pittsburgh Coma Scale,lesions region and volume were analysed among the various stroke types groups.Results The levels of serum NSE in the stroke patients at acute stage were significantly higher than that in NC group (allP