Preliminary Application Of SELDI-TOF-MS In Congenital Defect

Background and Purpose:Congenital defect is a widely-used term for a congenital malformation, including structural abnormalities,functional defects,metabolic disorders,genetic abnormalities and behavioral aspects of all the anomalies.In a variety of human congenital defects,their etiology and pathogenesis have not yet completely clear, but the epidemiology,genetics,molecular biology studies suggest the direct cause was genetic factors,environmental factors or the interaction of genetic factors and environmental factors.For 65-70%of anomalies there seems to be a "multi factorial" cause,meaning a complex interaction of genetic factors and environmental factors. For 20%-25%of anomalies have a purely genetic cause,only 10%of anomalies have a purely environmental cause(including physical factors,biological factors, chemical factors and nutritional factors and else).According to statistics,the incidence of congenital developmental defects of the total number of births was 2% -7%,nervous system malformations and chromosome abnormalities were the two most common defects.Reduce congenital defects,improve the quality of newborn babies are human mutual concern.Due to our country large population base and large birth population,congenital defects have become an important affecting factor to our national economic development and people normal life.Improve the quality of newborn babies is the prerequisite and foundation of improve the quality of the Chinese population,and promote China's economic prosperity and social stability development. Prenatal diagnosis,also known as intrauterine diagnosis or antenatal diagnosis,is employs a variety of techniques to determine the health and condition of an unborn fetus,determining the diagnosis of certain congenital,hereditary diseases,planning for selective abortion,or intrauterine treatment,reducing the birth rate of birth defects children and achieve the purpose of eugenics.Prenatal diagnosis is a major step forward in modem medical science and an important part of clinical obstetrics.In recent years,with the rapid progress of medical imaging,cell genetics, biochemistry and molecular biology,prenatal diagnosis has constantly enhanced.However,the current diagnostic methods are mainly limit to the discovery of fetal anomalies and reduce these the incidence of birth defects,rather than study a variety of abnormal fetal biological changes,pathogenesis,disease-related molecular markers,drug targets and else,it is imperative to select an appropriate method to inspect a variety of fetal abnormalities biological changes,pathogenesis, disease-related molecular markers,drug targets,screening of high sensitivity and specificity of fetal abnormalities markers.The emergence of proteomics has brought hope and the dawn for us to overcome this problem.Proteomics is the large-scale study of proteins,including all expressed proteins of cell,tissue or body fluids.Proteomics research the composition and the law of all protein encoded by genome in different time and space,the dynamic description of gene regulation,the quantitative study of proteins,the disease and drug impact to life processes,as well as the interpretation of gene expression and regulation mechanism.The cores of proteomics research is compare the common pathology individuals and search the specific disease-related protein molecules,in order to explore the pathogenesis,design new drugs for the disease,and provide the molecular level experimental and theoretical basis for early diagnosis and prognosis prediction.As genomics research development,proteomics research has become an extremely active part of the world's life sciences,the core of the functional genomics era or "post-genome era",and a powerful tool of the biomedical field.Proteomic technology is only recently beginning to be employed in pregnancy research.Although proteomics study is not widely used in reproductive medicine, but the emergence of proteomics research added a new research technology and a new research dimension to the field of reproductive medicine.Application of these high throughput methodologies in pregnancy-related pathology has contributed to the comprehension of the underlying pathophysiologies and the successful identification of relevant protein biomarkers that can potentially change early diagnosis and treatments of several medical conditions related to human pregnancy and significantly improve maternal health.Currently,proteomics research is mainly using the traditional proteomics techniques such as two-dimensional gel electrophoresis,mass spectrometry,but these technologies are not suitable for a large scale difference proteome study because of their high request,complicated procedure,time consuming,et al. Surface-enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS) which convergence the two major advantages of mass spectrometry and chip technology is a new proteomics research methods in recent years,which has numerour advantages,such as high-throughput,high sensitivity and eles.It has the ability of identification of potential biomarkers for a variety of diseases through overall view analysis in samples and provided a brand-new technology platform for early diagnosis and treatment efficacy monitoring tool of diseases.Amniotic fluid is the nourishing and protecting liquid contained by the amnion of a pregnant woman and an important and indispensable element for maintenance fetal life.The magic water can be the potential and rich biomarkers source of maternal and fetal disease.Based on the current traditional maternal serum prenatal screening,ultrasound screening for fetal malformation,ultrasound-guided puncture the amniotic cavity,amniotic cell culture,chromosome karyotype analysis,this study has been used SELDI-TOF-MS to study amniotic fluid proteomics of normal fetuses and chromosome abnormalities fetuses,normal fetuses and nervous system malformations fetuses,to establish a normal fetal amniotic fluid proteomic fingerprint / database,to study the amniotic fluid protein expressed difference between physiological and pathological state of fetuses by using proteomics, genomics and bioinformatics technologies,to search specific protein molecules for abnormal fetuses,to explore etiology,pathogenesis,pathophysiological changes in protein levels of chromosome abnormalities fetuses and nervous system malformations fetuses,to provide molecular biomarker for early diagnosis of abnormal fetuses,to avoid a heavy burden to families and society by timely prenatal diagnosis of chromosome abnormalities fetuses and nervous system malformations fetuses prevention,to provide new molecular targets for designed new prevention and treatment drugs.Partâ… Detection of Distinct Protein in Amniotic Fluid of Chromosome Abnormalities fetuses by SELDI-TOF-MS TechnologyObjective To detect the distinct protein in amniotic fluid between chromosome abnormalities fetuses and chromosome normal fetuses.Methods Surface-enhanced laser desorption-ionization/time-of-flight mass spectrometry was used to characterize amniotic fluid peptides in amniotic fluid between chromosome abnormalities fetuses and normal fetuses.WCX2 protein chips were used to characterize amniotic fluid peptides in amniotic fluid.Protein chips were examined in PBSâ…¡C protein reader,the protein profiling was collected by proteinchip software 3.1 and analyzed by Biomarker Wizard software.Results 5 distinct proteins were identified in amniotic fluid between chromosome abnormalities fetuses and normal fetuses.Compared with control group,three proteins with m/z 4967.526Da,5258.056 Da,6868.197 Da were down-regulated,and two proteins with m/z 4134.51 Da,5423.397 Da up-regulated in chromosome abnormalities fetuses.Conclusion Proteomic analysis was able to detect distinct proteins in protein profiling of amniotic fluid between chromosome abnormalities fetuses and normal fetuses. Objective To detect the distinct protein in amniotic fluid between nervous system malformations fetuses and normal fetuses.Methods Surface-enhanced laser desorption-ionization/time-of-flight mass spectrometry was used to characterize amniotic fluid peptides in amniotic fluid between nervous system malformations fetuses and normal fetuses.WCX2 protein chips were used to characterize amniotic fluid peptides in amniotic fluid.Protein chips were examined in PBSâ…¡C protein reader,the protein profiling was collected by proteinchip software 3.1 and analyzed by Biomarker Wizard software.Results 9 distinct proteins were identified in amniotic fluid between nervous system malformations fetuses and normal fetuses.Compared with control group,three proteins with m/z 4967.5Da,5258.0 Da,11717.0 Da were down-regulated,and six proteins with m/z 2540.4 Da,3107.1 Da,3396.8 Da,4590.965 Da,5589.2 Da,6429.4 Da up-regulated in nervous system malformations fetuses.Conclusion:Proteomic analysis was able to detect distinct proteins in protein profiling of amniotic fluid between nervous system malformations fetuses and normal fetuses.Conclusion1.This study is a multidisciplinary,interdisciplinary and comprehensive research coordinate with medical imaging,ultrasound intervention,genetic,basic medicine, obstetrics.The work flow of prenatal screening,prenatal diagnosis of fetal abnormality in the second trimesters had laid a solid foundation for the preliminary application of SELDI-TOF-MS in prenatal diagnosis.2.SELDI-TOF-MS was a stable and reliable technology for fetal amniotic fluid protein expression with good repeatability and can be an effective method for the fetal amniotic fluid proteomics. 3.This study preliminarily established the amniotic fluid proteomic fingerprint / database of normal fetuses,chromosome abnormalities fetuses,nervous system malformations fetuses with the application of SELDI-TOF-MS SELDI technology.4.Five proteins peaks M / z 4134.51 Da,4967.526Da,5258.056 Da,5423.397 Da, 6868.197 Da could be specific biomarkers for chromosome abnormalities fetuses.5.Nine proteins peaks M / z 2540.4 Da,3107.1 Da,3396.8 Da,4590.965 Da, 4967.5 Da,5258.0 Da,5589.2 Da,6429.4 Da,11717.0 Da could be specific biomarkers for nervous system malformations fetuses.6.SELDI-TOF-MS technique has a broad application prospects in prenatal diagnosis.Main Innovative Points1.Based on a complete,scientific,effective work flow for congenital defect in the second trimesters,this study first used SELDI-TOF-MS to study amniotic fluid proteomics of fetuses for prenatal diagnosis.2.SELDI-TOF-MS technology application preliminarily established amniotic fluid proteomic fingerprint / database of normal fetuses,chromosome abnormalities fetuses and nervous system malformations fetuses.3.This study was the first to investigate the value of proteomic in the diagnosis of fetal chromosomal abnormalities,and five proteins could be specific biomarkers for chromosome abnormalities fetuses.4.This study was the first to investigate the value of proteomic in the diagnosis fetal nervous system malformations,and nine proteins could be specific biomarkers for nervous system malformations fetuses.