Two-dimensional electrophoretogram of acute brain injury-associated proteins Comparison between Inju

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BACKGROUND:To this date,specific molecular markers for early diagnosis and prognosis monitoring ofcraniocerebral injury in clinical medicine do not exist.Therefore,differential detection of specific proteinsmight play an important role in diagnosis and treatment of this type of brain injury.OBJECTIVE:To compare differential cerebral cortical protein expression of craniocerebral injury patientsand normal subjects through the use of proteomics.DESIGN:Contrast observation.SETTING:Department of Neurosurgery,Xiangya Hospital of Central South University.PARTICIPANTS:Ten patients(6 males and 4 females,20-58 years old),with severe craniocerebral injury,were selected at the Department of Neurosurgery,Xiangya Hospital of Central South University,from June2004 to December 2006.All patients were diagnosed with CT test and Glasgow test(scores <8).Surgery was performed 4-12 hours after craniocerebral injury,and injured cortical tissues of the frontal and temporal lobes were resected for sampling.At the same time,control cortical tissues were collected from frontal and temporal lobes of 2 epileptic patients who underwent hippocampus-nucleus amygdala resection,and 2 lateral ventricular tumor patients who underwent tumor resection.The participants and their relatives provided confirmed consent,and this study received confirmed consent from the local ethics committee. METHODS:Ten samples from injured patients and 4 normal samples were compared through the use of proteomics.Total protein was separated by using two-dimensional electrophoresis with immobilized pH gradients,and the differential protein expressions were compared using image analysis after blue-sliver staining. Differential protein spot expressions were analyzed with a matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI/TOF MS) and electrospray ionization-quadrupole time of flight mass spectrometry(ESI-Qq TOF MS).MAIN OUTCOME MEASURES: ①Two-dimensional electrophoresis of protein from cerebral cortex; ②differential protein expression. RESULTS:①Two-dimensional electrophoresis of protein from cerebral cortex:two-dimensional gel electrophoretogram,which is considered to have high resolution and consistent duplication,was performed on injured cortical tissues and normal cortical tissues.The image analysis system detected 21 differential protein pots.②Differential protein spot expressions:mass spectrometry resulted in 17 differential protein spots that related to metabolic response,oxidative stress response,and signal transduction.CONCLUSION:MALDI/TOF MS and ESI-Qq TOF MS are exceptional methods for evaluating differential protein expression.Results from this study indicated 17 different craniocerebral injury-associated proteins.
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