基于UPLC-Q/TOF和1HNMR的代谢组学技术在乳腺癌步研究中的应用
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摘要
实验目的:
初步建立基于1HNMR技术和的UPLC-Q/TOF(液质联用)技术代谢组学方法,并将该方法应用于乳腺癌患者血清中内源性代谢物的变化,寻找乳腺癌相关诊断标志物。在此基础上初步比较了化疗方案AC(阿霉素+环磷酰胺)干预前后乳腺癌患者血清中代谢物的变化,观察化疗方案干预后患者体内血清中内源性代谢物的变化趋势。
研究方法:
1.研究对象的纳入:选取体检结果正常的11例健康人作为正常对照组;选取符合纳入标准的16例初诊乳腺癌患者作为实验组。
2.标本采集:采集11例正常女性静脉血,分离并收集血清;分别采集实验组16例乳腺癌患者在化疗前及化疗一个疗程后的静脉血,分离并收集
3.建立基于1HNMR技术的血清代谢组学分析方法,包括实验方法和模式识别方法的选择。并将建立的方法正常女性和实验组化疗前血清的代谢组学分析,找出血清中代谢物的差异;同时将方法应用于AC方案干预前后乳腺癌患者血清中的代谢物变化。通过文献检索和数据查询找出血清中差异。
4.建立基于液质联用技术(UPLC-Q/TOF)技术的血清代谢组学分析平台,并将建立的方法正常女性和实验组化疗前血清的代谢组学分析,找出差异;同时将方法应用于AC方案干预前后乳腺癌患者血清中的代谢物变化。和1HNMR技术研究结果进行比较。
5.数据分析:通过多种分析统计分析方法对产生的海量数据进行分析,分析过程包括:归一化、修正80%规则、数据集分割和数据缩放等方法对数据集进行预处理;通过数据分析:采用主成分分析(principal componentanalysis,PCA)、正交偏最小二乘判别分析(OrthogonalPartial Least-Squares Discriminant Analysis,OPLS-DA)进行模式识别分析;根据模型的变量重要性因子(VIP值)、非参数检验结果和P值筛选潜在标记物。
实验结果:
1.建立了1H-NMR的血清代谢组学方法,健康女性和乳腺癌患者化疗前、化疗方案干预前后两个组经代谢组学方法得到很好的区分,并找到了相对应的16个乳腺癌肿瘤标志物。乳酸(lactate,1.33、4.12),缬氨酸(valine,0.99/1.04),N-乙酰糖蛋白(N-acetylgly-coproteins,2.04),谷氨酰胺(glutamine,2.41),磷酸胆碱/胆碱(phosphocholine/cholie,3.22),葡萄糖(a-glucose和B-glucose,3.40-3.90,4.66,5.22),酪氨酸(tyrosine,6.87,7.17),组氨酸(histidine), LDL/VLDL,不饱和脂肪酸等峰有增大或减小的表现。
2.建立了UPLC-Q/TOF-MS的血清代谢组学研究方法,发现胆碱类和苯丙氨酸、异亮氨酸在正常人和乳腺癌患者化疗前有不同,其中化疗后的乳腺癌患者血清中的胆碱类有向正常变化单位趋势。1HNMR技术所得到的标记物和UPLC-Q/TOF得到的标记物大部分不同,但是分组聚类结果类似。
结论:
~1HNMR技术和UPLC-Q/TOF技术作为常用的代谢组学分析工具,在标记物发现中有一定的互补性。通过对正常人和乳腺癌患者化疗前后的代谢组学研究,得到了三者的代谢谱,发现本文所建立的NMR技术和UPLC-Q/TOF技术为基础的代谢组学方法应用于乳腺癌患者血清中具有良好的聚类效果。
Objective:
~1HNMR and UPLC-Q/TOF based metabolic profiling was used to investigate the differences of serum metabolic patterns between patients with Breast cancer and healthycontrols to find tentative diagnostic biomarkers.Based on the findings above,we tried to describe the metabolite changes related to the continuous phases ofradiotherapy.It will provide a novel technological platform for the mechanism study of Breast cancer.
Methods:
1.Patients:16 newly diagnosed patients with Breast cancer who match the selection conditions served as Breast cancer group.11 healthy persons served as control group.
2.Sample collection:Serum samples were collected from 11 persons in the control group through venous blood.Serum samples of 16 patients in the Breast cancer group were collected before chemotherapy and two weeks after chemotherapy.
3.NMR analysis of serum samples:The NMR technique based metabonomic method was established,and the serum was introduced to NMR.Applying the NMR metheod,the serum samples from the health and Breast cancer were collected and detected by the machine. The serum endogenous metabolites were identified.
4. Eatabilished the approach with Ultra-performance-Liquid Chromatography- ElectroSpray Ionization-quatrupole-Time Of Flight mass spectrometry (UPLC-ESI-Q-TOF)for metabonomic study,and introduced the method to the collecded serum to judge the state of disease.
5. Data analysis:Then the data was imported SIMCA-P+ software for denoising. Through a variety of analysis of statistical analysis method of analysis produced massive data,analyzing process including: through the normalization, correction 80% rules, data set and data zoom in a set of data pretreatment method; Through theanalysis of data: principal by principal component analysis (PCA), slant component analysis, the least squares discriminant analysis (discriminant squares - order extra transportation charges nothing, PLS - DA) two methods of the samples classification; According to the model of variable importance factor (VIP value) and non-parameter test results and z value screening potential markers.
Results:
1. Eatabilished the approach with NMR for metabonomic study, some specific serum endogenous metabolites changes in the metabolic composition of serum samples from Breast cancer,there are lactate,valine, N-acetylgly-coproteins, glutamine,phosphocholine/choline, a-glucose/B-glucose,tyrosine,histidine,LDL/VLDL.
2. Eatabilished the approach with for metabonomic study.Examination of the metabonomic data obtained by Ultra-performance-Liquid Chromatography -ElectroSpray Ionization-quatrupole-Time Of Flight mass spectrometry (UPLC-ESI-Q-TOF) ,the acquired metabonomic data was analyzed by multivariate analysis.Significant difference in endogenous metabolite profilea was observed in the control and patients with chemtheropy before and after.
Conclusions:
The relusts demonstrated that metabonomics is a potential and powerful tool for characterization of the metabolic perturbation under different physiopathologic status,and also indicated that the metabonomic analysis is promising to provide an integrative criterion to evaluate the severity of the Breast cancer and the prognosis.
初步建立基于1HNMR技术和的UPLC-Q/TOF(液质联用)技术代谢组学方法,并将该方法应用于乳腺癌患者血清中内源性代谢物的变化,寻找乳腺癌相关诊断标志物。在此基础上初步比较了化疗方案AC(阿霉素+环磷酰胺)干预前后乳腺癌患者血清中代谢物的变化,观察化疗方案干预后患者体内血清中内源性代谢物的变化趋势。
研究方法:
1.研究对象的纳入:选取体检结果正常的11例健康人作为正常对照组;选取符合纳入标准的16例初诊乳腺癌患者作为实验组。
2.标本采集:采集11例正常女性静脉血,分离并收集血清;分别采集实验组16例乳腺癌患者在化疗前及化疗一个疗程后的静脉血,分离并收集
3.建立基于1HNMR技术的血清代谢组学分析方法,包括实验方法和模式识别方法的选择。并将建立的方法正常女性和实验组化疗前血清的代谢组学分析,找出血清中代谢物的差异;同时将方法应用于AC方案干预前后乳腺癌患者血清中的代谢物变化。通过文献检索和数据查询找出血清中差异。
4.建立基于液质联用技术(UPLC-Q/TOF)技术的血清代谢组学分析平台,并将建立的方法正常女性和实验组化疗前血清的代谢组学分析,找出差异;同时将方法应用于AC方案干预前后乳腺癌患者血清中的代谢物变化。和1HNMR技术研究结果进行比较。
5.数据分析:通过多种分析统计分析方法对产生的海量数据进行分析,分析过程包括:归一化、修正80%规则、数据集分割和数据缩放等方法对数据集进行预处理;通过数据分析:采用主成分分析(principal componentanalysis,PCA)、正交偏最小二乘判别分析(OrthogonalPartial Least-Squares Discriminant Analysis,OPLS-DA)进行模式识别分析;根据模型的变量重要性因子(VIP值)、非参数检验结果和P值筛选潜在标记物。
实验结果:
1.建立了1H-NMR的血清代谢组学方法,健康女性和乳腺癌患者化疗前、化疗方案干预前后两个组经代谢组学方法得到很好的区分,并找到了相对应的16个乳腺癌肿瘤标志物。乳酸(lactate,1.33、4.12),缬氨酸(valine,0.99/1.04),N-乙酰糖蛋白(N-acetylgly-coproteins,2.04),谷氨酰胺(glutamine,2.41),磷酸胆碱/胆碱(phosphocholine/cholie,3.22),葡萄糖(a-glucose和B-glucose,3.40-3.90,4.66,5.22),酪氨酸(tyrosine,6.87,7.17),组氨酸(histidine), LDL/VLDL,不饱和脂肪酸等峰有增大或减小的表现。
2.建立了UPLC-Q/TOF-MS的血清代谢组学研究方法,发现胆碱类和苯丙氨酸、异亮氨酸在正常人和乳腺癌患者化疗前有不同,其中化疗后的乳腺癌患者血清中的胆碱类有向正常变化单位趋势。1HNMR技术所得到的标记物和UPLC-Q/TOF得到的标记物大部分不同,但是分组聚类结果类似。
结论:
~1HNMR技术和UPLC-Q/TOF技术作为常用的代谢组学分析工具,在标记物发现中有一定的互补性。通过对正常人和乳腺癌患者化疗前后的代谢组学研究,得到了三者的代谢谱,发现本文所建立的NMR技术和UPLC-Q/TOF技术为基础的代谢组学方法应用于乳腺癌患者血清中具有良好的聚类效果。
Objective:
~1HNMR and UPLC-Q/TOF based metabolic profiling was used to investigate the differences of serum metabolic patterns between patients with Breast cancer and healthycontrols to find tentative diagnostic biomarkers.Based on the findings above,we tried to describe the metabolite changes related to the continuous phases ofradiotherapy.It will provide a novel technological platform for the mechanism study of Breast cancer.
Methods:
1.Patients:16 newly diagnosed patients with Breast cancer who match the selection conditions served as Breast cancer group.11 healthy persons served as control group.
2.Sample collection:Serum samples were collected from 11 persons in the control group through venous blood.Serum samples of 16 patients in the Breast cancer group were collected before chemotherapy and two weeks after chemotherapy.
3.NMR analysis of serum samples:The NMR technique based metabonomic method was established,and the serum was introduced to NMR.Applying the NMR metheod,the serum samples from the health and Breast cancer were collected and detected by the machine. The serum endogenous metabolites were identified.
4. Eatabilished the approach with Ultra-performance-Liquid Chromatography- ElectroSpray Ionization-quatrupole-Time Of Flight mass spectrometry (UPLC-ESI-Q-TOF)for metabonomic study,and introduced the method to the collecded serum to judge the state of disease.
5. Data analysis:Then the data was imported SIMCA-P+ software for denoising. Through a variety of analysis of statistical analysis method of analysis produced massive data,analyzing process including: through the normalization, correction 80% rules, data set and data zoom in a set of data pretreatment method; Through theanalysis of data: principal by principal component analysis (PCA), slant component analysis, the least squares discriminant analysis (discriminant squares - order extra transportation charges nothing, PLS - DA) two methods of the samples classification; According to the model of variable importance factor (VIP value) and non-parameter test results and z value screening potential markers.
Results:
1. Eatabilished the approach with NMR for metabonomic study, some specific serum endogenous metabolites changes in the metabolic composition of serum samples from Breast cancer,there are lactate,valine, N-acetylgly-coproteins, glutamine,phosphocholine/choline, a-glucose/B-glucose,tyrosine,histidine,LDL/VLDL.
2. Eatabilished the approach with for metabonomic study.Examination of the metabonomic data obtained by Ultra-performance-Liquid Chromatography -ElectroSpray Ionization-quatrupole-Time Of Flight mass spectrometry (UPLC-ESI-Q-TOF) ,the acquired metabonomic data was analyzed by multivariate analysis.Significant difference in endogenous metabolite profilea was observed in the control and patients with chemtheropy before and after.
Conclusions:
The relusts demonstrated that metabonomics is a potential and powerful tool for characterization of the metabolic perturbation under different physiopathologic status,and also indicated that the metabonomic analysis is promising to provide an integrative criterion to evaluate the severity of the Breast cancer and the prognosis.
引文
[1] J.K.Nicholson, J.C.Lindon,E.Holmes. Metabonomics: understanding the Metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data[J]. Xenobiotica, 1999, 29(11): 1181-1189.
[2] J.K. Nicholson, J. Connelly, J.C. Lindon, et al . Metabonomics : a platform for studying drug toxicity and gene function [J]. Nat Rev Drug Disc,2002, 1 (2) :153-161.
[3] O.Fiehn . Combining genomics, metabolome analysis, and biochemical modeling to understand metabolic networks [J] .Comp Funct Genomics ,2001, (2): 155-168.
[4] J.K.Nicholson,I.D.Wilson. Opinion:understanding‘global’systems biology:Mera-bonomies and the continuum of metabolism.Nat Rev Drug Discov,2003,2:668-676.
[5] B.R.Baggett, J.D.Cooper , E.T. Hogan, et al . Profiling is of lavonoids found in legume root extracts using capillary electrophoresis[J]. Electrophoresis, 2002, 23 (11) : 1642-1651.
[6] J.L .Griffin , H.J.Williams ,et al. Metabolic profiling of genetic disorders : a multitissue 1H nuclear magnetic resonance spectroscopic and pattern recognition study into dystrophic tissue[J] . Anal Bio chem ,2001 ,293 (1) :16-21.
[7] Pelander,Ojanperal I,Lakes S.Toxicological screening with formula-based metabolite identification by liquid chromatography/ time-of-flight mass spectrometry [J] . Anal Chem ,2003 ,75(2):5710 -5718.
[8] Peiyuan Yina, Patamu Mohemaitib,et al.Serum metabolic profiling of abnormal savda by liquid-chromatography/mass-spectrometry[J].Journal of Chromatography B,2008 ,871(2):322-327.
[9] Bethanne, Warracka, et al. Normalization strategies for metabonomic analysis of urine samples[J]. Journal of Chromatography B, 2009,877(3): 547-552.
[10] Masahiro Sugimoto, David T. Wong, Akiyoshi Hirayama, etal.Capillary electrophoresis mass spectrometry-based saliva metabolomics identified oral, breast and pancreatic cancer-specific Profiles[J]. Metabolomics, 2010,6(1): 78-95.
[11] Kitano H. Systems biology: a brief overview[J]. Science, 2002,295 (5560) : 1662-1664.
[12] J.K.Nicholson, J.C.Lindon,HolmesE. Metabonomics: understanding the Metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data[J]. Xenobiotica.1999;29(11):1181-1189.
[13] FiehnO. Combining genomics, metabolome analysis, and biochemical modeling understand metabolic networks [J] .Comp Funct Genomics ,2001,2 :155-168.
[14]朱勇飞,张天宝.代谢组学在毒理学研究中的应用[M].国外医学:卫生学分册,2005 ,32(3) :156 - 159.
[15]杨军,宋硕林,Jose CP等.代谢组学及其应用[J].生物工程学报,2005,1(21):1-5.
[16]许国旺,杨军.代谢组学及其研究进展[J].色谱,2003,1(21): 316-320.
[17] C.W.Mun, J.Y.Cho, W.J.Shin.et a1.Exvivo proton MR spectroscopy(1H-MRS) for evaluation of human gastric carcinoma[J].Magn Reson Imaging, 2004, 22(6):861-870.
[18] Katz-Brull R, Seger.D, Rivenson-Segal.D, et a1. Metabolic markers of Breast cancer enhanced choline metabolism and reduced choline-ether-phospholipid synthesis[J].Cancer Res, 2002, 62(7) :1966-1970.
[19] Heintz P, Ehreneim Ch, Hundeshagen H. In Application of NMR Techniques on the Body Composition of Live Animals[EB/OL]. London, Elsevier, 1989 .
[20] Clare A, Daykin, Peta J.D. Foxall, Susan C.Connor, et al.The Comparison of Plasma Deproteinization Methods for the Detection of Low-Molecular-Weight Metabolites by 1H Nuclear Magnetic Resonance Spectroscopy[J]. Analytical Biochemistry ,2002,304(7):220-230.
[21] J. K. Nicholson , J. D . Foxall. 750 MHz IH and IH-l3C NMR Spectroscopy of Human Blood Plasma[J]. Anal.Chem, 1996,67(5):793-811.
[22] Kunle Odunsi,Robert M. Wollman,Christine B. Ambroson, Detection of epithelial ovarian cancer using 1H-NMR-based metabonomics[J], Int. J. Cancer, 2005,113(5):782-788.
[23]艾斯克尔.吐拉洪,哈木拉提.吾甫尔,豪富华.基于NMR的维吾尔医异常黑胆质型肿瘤患者血浆代谢组学分析[J],科技导报,2009,27(13):27-31.
[24]陆强,黄一红,丛辉等.肝细胞癌、肝硬化患者血清中代谢物组研究[J],分析化学,2009,39(2): 194-198.
[25]姜玉章,孙晓阳,史昆波,食管鳞癌患者血清1H NMR指纹图谱研究[J].南京医科大学学报,2009,29(11):1560-1563.
[26] H.J.Roeijmans, J.D.De Hoog, C.S.Tan. et al. Molecular taxonomy and GC/MS of metabolites of Scytalidum hyalinum and Nattrassia mangiferae (Hendersonula tofllloidea)[J], Med Vet Mycol, 1997, 35(3): 181-189.
[27] D.V.Huhman, L.W.Sumner. Metabolioc profiling of saponin glycosides in Medicago sativa and Medicago truncatula using HPLC coupled to an electrospray ion trap mass spectometer [J]. Phytochemistry, 2002, 59 (3) : 347-360.
[28] Soga T, Ueno Y, Naraoka H, et al.Simultaneous determination of anionic intermediates for bacillus subtilis metabolic pathways by capillary electrophoresis electros pray ionization mass spectrometry[ J ]. Anal Chem, 2002, 74 (10) : 2233-2239.
[29] H.J.Major, R.Williams, A.J.Wilson, I.D.Wilson. A metabonomic analysis of plasma from Zucker rat strains using gas chromatography/mass spectrometry and pattem recognition[J]. Rapid Commun Mass Spectrom ,2006,20(22): 3295-3302.
[30] D.S.Wishart, A.C.Guo, et al. HMDB: a knowledgebase for the human metabolome[DB]. Nucleic Acids Res. 2009 Jan; 37(Database issue): D603-610.
[31] J. T .Brindle, J. K. Nicholson, P. M.Schofield,et al. Application of chemometrics to 1H NMR spectroscopic data to investigate a relationship between human serum metabolic profiles and hypertension[J]. Analyst, 2003, 128(1):32-36.
[32] J.T..Brindle, H.Anni, E.Holmes, G.Tranter, J.K.Nicholson, H.W. Bethell, S .Clarke, P. M.Schofield, E. McKilligin, D. E. Mosedale, D. J. Grainger, Nat. Med. 2002, 8:1439-1442. [33 ] Ramadan Z, Jacobs D, Grigorov M, Kochhar S: Metabolic profiling using principal component analysis, discriminant partial least squares, and genetic algorithms[J]. Talanta, 2006, 68(5):1683-1691
[34] Odunsi K, Wollman RM, Ambrosone CB, Hutson A, McCann SE, Tammela J, Geisler JP, Miller G, Sellers T, Cliby W, Qian F, Keitz B, Intengan M, Lele S, Alderfer JL. Detection epithelial ovarian cancer using 1H-NMR-based metabonomics[J]. Int J Cancer, 2005, 113(5): 782-788.
[35] B.M. Beckwith-Hall, J.T .Brindle, R.H .Barton,et al. Application of orthogonal signal correction to minimise the effects of physical and biological variation in high resolution 1H NMR spectra of biofluids[J]. Analyst. 2002, 127:1283-1288.
[36] Holmes E,Antti H,Chemometric contributions to the evolution of metabonomics: mathematical solutions to characterising and interpreting complex biological NMR spectra[J]. Analyst, 2002, 127(12):1549-1557
[37] Millis K,Weybright P, CampbeilL N, et al . Classification of human liposarcoma and lipoma using exvivo proton NMR spectroscopy[J]. Magn Res on Med, 1999, 41 (2) : 257-267.
[38] J.T .Brindle,Antti H, Holmes E, et a1.Rapid and non invasive diagnosis of the presence and severity of coronary heart disease using 1H-NMR-based metabonomics[J]. Nat Med,2002,8(12):1439-1444.
[39] K.P.Chiang, Niessen S, Saghatelian A, et al. An enzyme that regulates ether lipid signaling pathways in cancer annotated by multi-dimensional profiling[J]. Chem Biol, 2006, 13 (10) : 1041-1050.
[40] J.L .Griffin , R.A. Kauppinen. A metabolomics perspective of human brain tumours [J] . FEBSJ , 2007 , 274 ( 5 ) : 1132 -1139.
[41] C.L.Florian, N.E.Preece, K.K.Bhakoo. Characteristic metabolic profiles revealed by 1 HNMR spectroscopy for three types of human brain and nervous system tumours[J].NMR Biomed , 995, 8(6) : 253 - 264.
[42] Odunsi K, R.M.Wollman, Ambrosone. Detection of epithelial ovarian cancer using 1 H-NMR-based metabonomics[J] . Cancer ,2005 ,113 (5) :782-788.
[43] Yang J. Diagnosis of liver cancer using HPLC-based metabonomics avoiding false-positive result from hepatitis and hepatocirrhosis diseases [J]. Chromatogr B Analyt Technol Biomed Life Sci ,2004 ,813 (1-2) :59- 65.
[44] Yang J , Xu G, Zheng Y. Strategy for metahonomics researchbased on high-performan liquid chromatography and liquidchromatography coupled with tandem mass spectromet ry[J ] .Chromatogr A ,2005 ,1084 (1 -2) : 214 - 221.
[45]陆强,黄一红,丛辉等.肝细胞癌、肝硬化患者血清中代谢物组研究[J],分析化学, 2009, 39(2):194-198.
[46]吴逸明等.高效液相色谱法测定肺癌患者尿中假尿嘧啶核苷和肌苷量比值[J].郑州大学学报(医学版)2002,37(4):433-435.
[47] Yan SK, Wei BJ, Lin ZY, et al. A metabonomic approach to the diagnosis of oral squamous cell carcinoma,oral lichen planus and oral ieukoplakia [J]. Oral Oncology, 2008, 44(5): 477-483.
[48]姜玉章,孙晓阳,史昆波.食管鳞癌患者血清1H NMR指纹图谱研究[J].南京医科大学学报,2009,29(11):1560-1563.
[49] Kubom A,M M Meguid,D C Hitch. Amino acid profiles correlate diagnostically with organ site in three kinds of malignant tumors[J]. Cancer, 1992,69(9): 2343-2348.
[50]汪晋,郁兰芳,沈朋,王书芳.气相色谱-质谱法分析乳腺癌患者血清代谢组[J],浙江大学报, 2009, 38(5): 45-48.
[51]何永频,陆瑞芳,吴岷.血清游离多不饱和脂肪酸的组分变化与乳腺癌的关系[J],中国临床营养杂志, 1999, 6(1): 21-23.
[52] Xu G, H.R.Schmid, ,et al. Excretionpattern investiation ofurinary normal and modified nucleosides of breas tcance rpatients by RP--HPLC andfaetoranalysis method[J]. Biomed Chromatogr, 2000, 14(7):459-466.
[53] Max Parkin,Freddie Bray,J.Ferlay,etal,Global Cancer Statistics,2002.CA Cancer ,2005,55:74-108
[54]中国乳腺癌流行病学调研项目[OL]: http://www.qgjk.org/html/2368.html.2011-03-25.
[55]李树云.触诊在乳癌诊断中的作用[J].江西医学院学报, 2002, 12( 5) : 48.
[56]于泽平,李幼生,王少华等.吸针穿刺吸取细胞学检查对乳腺癌的诊断价值[J].医学研究生学报, 2006,19(5) : 474-475.
[57]陈利平,乳腺癌发病机理的认识与治疗.2007亚太国际肿瘤生物学和医学学术会议暨首届解放军总医院肿瘤综合防治高峰论坛及第二届中国中青年肿瘤专家论坛论文集[C],2007.
[58]杨举伦,王丽.乳腺癌的分子病理学研究[J].临床与实验病理学杂志, 2011, 27(1): 10-14.
[59] Fattaneh A, T avassoli P D. Pathology and Genetics of the Breast and Female Genital Organs[ M ]. Lyon :I ARC Press , 2003: 49 -50.
[60]《乳腺癌HER2检测指南(2009版)》编写组.乳腺癌HER2检测指南( 2009版) [J].中华病理学杂志, 2009 , 38( 12) : 836-840.
[61]杨举伦,普苹,蔡学敏等.乳腺不典型增生组织中Hras基因突变的检测[J].临床与实验病理学杂志, 2001 , 17 ( 1 ) : 12-14.
[62]曹红婷,核磁共振代谢组学方法的体液制谱方法和制谱参数研究[D].厦门:厦门大学,2009:25-35.
[63] GoelA, J.R.anknecht. Concerted activation of ETS protein ER81 by p160 coactivators, the acetyltransferase p300 and the receptor tyrosine kinase HER2 /Neu[ J]. J Biol Chem, 2004, 279 ( 15) : 14909-14916.
[64]杨举伦,蔡学敏,普苹等.乳腺增生病P53基因第5外显子突变及其蛋白表达[J] .临床与实验病理学杂志, 2002 , 18( 3 ): 264-267.
[65]杨举伦, David K, Michael J B等.乳腺癌发生过程中NOEY2基因启动子区甲基化及mRNA表达[J] .临床与实验病理学杂志, 2005 , 21( 5 ) : 543-547.
[66] Yang J L , Ren Y, Wang L , et al . PTEN mutation spectrumin breast cancers and breast hyplasia[J]. J Cancer Res Clin Onco, 2010, 136( 9 ) : 1303-1311.
[67]杨海捷,王丽,魏万里等.散发性乳腺癌及乳腺不典型导管增生组织BRCA1基因启动子区甲基化分析[J] ,临床与实验病理学杂志, 2008 , 24( 2 ): 141- 145.
[68]谷永强,朱秋娟,肖俊英等. CA15-3和CEA检测在乳腺癌诊断中的意义[J],中国误诊学杂志, 2001, 1(11): 1619-1620.
[69]于鹏,杨兴无,杨春明等.乳腺癌手术前后肿瘤标志物CA153的检测及其临床意义[J],中华普通外科杂志, 2002, 17(8): 496.
[70]谢翠华,陆亚平. CA153在乳腺癌诊断和肿瘤分期中的应用[J],实用心脑肺血管病杂志, 2008, 16(11): 44-45.
[71]陈智周,范振符,杨剑等.肿瘤标志物CA153的免疫放射分析及其临床应用[J],中华肿瘤杂志, 1998, 20(2): 125-128..
[72] W.D.Dupont, F.F.Parl, W.H.Hartmann, et al. Breast cancer risk associated with proliferative breast disease and atypical hyperplasia[J]. Cancer, 1993, 71(4): 1258-1265.
[73]王鸿利.实验诊断学[M[.北京:人民卫生出版社, 2001: 240-242.
[74]朱海龙. CEA、AFP、CA125、CA15-3、CA19-9检测对恶性肿瘤的诊断价值探讨[J],现代医药卫生, 2006, 22(22): 3422-3423.
[75]朱波,黄文成,黄玲莎.血清CA153、CEA、TSGF联合检测在128例乳腺癌中的临床应用[J],福建医药杂志, 2003, 25(4): 155-174.
[76]王旭东.血清VEGF、CA153及CEA联合检测对乳腺癌的诊断价值[J],中国医师进修杂志, 2007, 30(4): 28-31.
[77]叶蓓,柳光宇,陆劲松.常用的乳腺癌血清肿瘤标志物在早期诊断中的应用价值不高[J],《中国癌症杂志》, 2009, 19(10): 807-808.
[78]沈坤炜,胡震.乳腺肿块的诊断和鉴别诊断[J].中国实用外科杂志,2005, 25(2):160-162.
[79]王颀,连臻强.乳腺癌早期诊断的临床思路[J],中华乳腺病杂志(电子版),2010,4(4):357-360.
[80] B.D.Ross, GK,Radda, D.G.Gadian,et a1. Preliminary observations on the metabolic responsesto exercise in humans, using 31-phosphorus nuclear magnetic resonance[J]. Ciba Found Symp,1982 ,87:145-152.
[81] Suna,SalnIiIlen,A.Soininen,P.Laatikamell,et al. l HNMR metabonomics of plasma lipoprotein subclasses:elucidation of metabolic clusteriIlng by self-oranising maps[J]. NMR Biomed, 2007, 20(7): 658-672.
[82] Akira K, Masu S, Imachi M, et al. 1HNMR-based metabonomic analysis of urine from young spontaneously hypertensive rats[J]. J Pham and Biomed Anal, 2008, 46(3): 550-556.
[83] Shi C, Wu CQ, Cao AM, et al. NMR-spectroscopy-based metabonomic approach to the analysis of bay41-4109, a novel anti-HBV compound,induced hepatotoxicity in rats[J]. Toxicol. Lett, 2007, 173(3):161-167.
[85] ] Liao PQ,Wei L,Zhang XY,et al. Metabolic pofiling of serum f rom gadolinium chloride Treated rats by 1HNMR spectroscopy[J]. Anal Biochem, 2007, 364(2):112-121.
[86] Ramadan Z, Jacobs D, et al. Metabolic Profiling using Principcal component diseriminant partial least squares,and genetic algoriths[J]. Talanta, 2006, 68(5):1683-1691.
[87] Liao PQ, Wei L, Zhang XY, et al. Metabolic profiling of serun from gadolinium chloride-treated Rats by 1HNMR,spectroscopy[J]. Anal Biotech, 2007, 364(2): 112-121.
[88] Wu HF, Zhang XY, Li XJ, et al. Comparison of metabolic profiles from serum from hepatotoxin-treted rats by nuclear-magnetic-resonnce-based metabonomic analysis [J]. Anal Biochem, 2005,340(1):99-105.
[89] Hewer R, Vorster, Steffens FE, et al. Applying biofluid 1HNMR-based metabonomic techniques to diatinguish between HIV-1 postive /AIDS patients on antiretroviral trentmentand HIV-1 negative individuala[J]. J Pham and Biomed Anal, 2006, 4(4): 1442-1446.
[90] J.T.Brindle, H.Anni, E.Holmes, J.K.Nicholson, H.W.Bethell, S.Clarke, P.M.Schofield, E .McKilligin, D.E. Mosedale, D. J. Grainger, Nat. Med. 2002, 8(11): 1439-1442.
[91] Ramadan Z, Jacobs D, Grigorov M, Kochhar S: Metabolic profiling using principal component analysis, discriminant partial least squares, and genetic algorithms[J]. Talanta ,2006, 68(5):1683-1691
[92] Odunsi K, R.M .Wollman, C.B. Ambrosone, Hutson A, S.E. McCann, Tammela J, J.P Geisler, Miller G, Sellers T, Cliby W, Qian F, Keitz B, Intengan M, Lele S, J.L .Alderfer. Detection epithelial ovarian cancer using 1H-NMR-based metabonomics[J]. Int J Cancer, 2005, 113(5): 782-788.
[93] Peiyuan Yin, Patamu Mohemaiti, Jing Chen, et al. Serum metabolic profiling of AbnormalSavda by liquid chromatography/mass spectrometry[J]. Journal of Chromatography B, 2008,43.(5): 1692-1698.
[94]冒海蕾,徐晏,王斌等.正交信号校正在正常人血清1H NMR谱的代谢组分析中的滤噪作用评价[J].化学学报, 2007, 65(2): 152-158.
[95]杨永霞,杨生义,梁敏锋.基于核磁共振波谱的乙肝患者血清代谢组研究[J],第二军医大学学报, 2010,31(3):388-291.
[96]阿仙姑·哈斯木,巴吐尔.买买提明,伊力亚尔.夏合丁.不同模式识别方法分析食管癌患者血浆代谢组的磁共振氢谱[J],中华肿瘤杂志, 2010, 32(9):681-684.
[97]高岗,杨根金,娄子洋.肾虚证大鼠尿液的核磁共振谱代谢组学研究[J].第二军医大学学报, 2009, 30: 565 - 568.
[98]艾斯克尔.吐拉洪,哈木拉提.吾甫尔,豪富华.基于NMR的维吾尔医异常黑胆质型肿瘤患者血浆代谢组学分析[J],科技导报,2009,27(13):27-31.
[99] C.W. Mun,J.Y. Cho,W.J .Shin, et a1. Ex vivo proton MR spectroscopy(1H-MRS) forevaluation of human gastric carcinoma[J].Magn Reson Imaging, 2004, 22(6):861-870.
[100] K.W .Jordan, L.L. Cheng. NMR-based metabolomics approach to target biomarkers for human prostate cancer [J]. Expert Rev Proteomics , 2007 , 4 (3): 389 - 400.
[101] K .Odunsi, R.M. Wollman, C.B.Ambrosone, et a1. Detection of epithelial ovarian cancer using 1 H-NMR-based metabonomics[J]. Int J Cancer, 2005, 113(5):782-788.
[102] J. E. Entire, K.C.Kuo, M.E.Smith, et a1. Classfication of lung-cancer patients and controls by chromatography of modified nucleosides in serum[J]. Cancer Res, 1989, 49(15):1057.
[103] Odunsi K, R.M .Wollman, C.B. Ambrosone. Detection of epithelial ovarian cancer using 1 H-NMR-based metabonomics[J]. 1nt J Cancer, 2005, 113 (5): 782 - 788.
[104] Denkert C , Budczies J , Kind T. Mass Specromery-based metabolic profiling reveals different metabolite patterns in invasive ovarian carcinomas and ovarian borderline tumors[J] . Cancer Res ,2006 ,66 (15): 10795-10804.
[105] Bethanne M.Warracka, Serhiy Hnatyshyna, Karl-Heinz Ott, Normalization strategies for metabonomicanalysisof urine samples[J]. Journal of Chromatography B, 2009, 877:547-552.
[106] S. Bijlsma, L.Bobeldijk, E.R..Verheij, et al. Large-scale human metabolomics studies: A strategy for data (pre-) processing and validation[J]. Anal Chem, 2006, 78(2): 567-574.
[107] G .B .Mills, W.H.Moolenaar. The emerging role of lysophosphatidic acid in cancer[J]. Nat Rev Cancer, 2003, 3(8): 582-591.
[108] P.Y .Yin, X. J .Zhao, Q .R,Li , J .S. Wang,et al. Metabonomics study of intestinal fistulas based on ultraperformance liquid chromatography coupled with Q-TOF massspectrometry (UPLC/Q-TOF MS)[J]. J Proteome Res, 2006, 5(9): 2135-2143.
[109] J.C.Lindon, E.Holmes, J.K.Nicholson. Metahonomics: Systems biology in pharmaceutical research and development[J]. Curr Opin Mol Ther, 2004,6(3): 265-272.
[110] User's Guide to SIMCA-P, SIMCA-P+ version 12.0.
[111]林青认,超高液相-质谱联用对胃癌代谢组学研究[D].杭州:浙江大学, 2010: 8-10.
[112] S.D .Hey, G.M.Park, M.A. MacnurLatl, et al. Stimulalation of protein synthesis in human tumors by parentetal nutrition:evidence for modulation of tumor growth[J]. Br J Surg, 1991, 78(4): 483-487.
[113]洪毅,谈冶雄,王红阳.代谢组学-肿瘤学研究的重要技术平台[J],中国肿瘤生物治疗杂志,2007,14(6):585-588.
[114] A.M .Proenza, Oliver J, Palou A, et al, Breast and lung cancer are associated with a decrease in blood cell aminoacid content[J]. J Nutr Biochem, 2003, 14(3):133-138.
[115]王喜,基于代谢组学的胃癌化疗敏感性和毒性预测[D].上海:第二军医大学, 2008:20-35.
[116] Cai H, Erhardt P. Hydrolysis of phosphatidylcholine couples Ras to activation of Raf protein kinase during mitogenic signal transduction[J]. Mol Cell Biol, 1993, 13(12): 7645-7651.
[117] Mulder C, L.O Wahlund, Decreased lysophosphatidylchoine/phosphatid ylcholine ratio in cerebrospinal fluid in Alzheimer's disease[J], J Neural Transm, 2003, 110(8): 949-955.
[118] Lee ES,Chen H,Inhibitory effects of lysophpsphatidylcholine on the dopaminergic system[J]. Neuro Chem Res, 2004, 29(7): 1333-1342.
[119] Y.L.Ting, D.Sherr. Variations in energy and Phospholipid metabolism in normal and cancer human mammary epithelial cells[J]. Anti caneer Res, 1996, 16(3B): 1381-1388.
[120] C.G.Gaetano, N.Samadi, Inhibition of autaxin prodution or activity blocks Lysophosphatidyl choline-induced migration of human breast cancer and mekanoma cells[J]. Mol Carcinog, 2009, 48(9): 801-809.
[121] D.N.Brindley, Hepatic secretion of lysphosphatidylcholine: A novel transport system for polyunsaturated fatty acids and choline[J]. J Nutr Biochem, 1993, 4: 442-449.
[122] E.Koh, R.W. Bandle, Novel point mutations attenuate autotaxin activity.Lipids [J]. Lipids Health Dis,2009,8:4
[123] Fang X, Schummer M, Lysophosphatidic acid is a bioactive meidtor in ovarian cancer[J], Biochim.biophys,actal, 2002, 82(15):254-264.
[124] Tarah L, Pua,Feng-qiang,Wang, Roles of LPA in ovarian cancer developmengt and proression[J], Future Oncol, 2009, 5(10):1649-1673
[2] J.K. Nicholson, J. Connelly, J.C. Lindon, et al . Metabonomics : a platform for studying drug toxicity and gene function [J]. Nat Rev Drug Disc,2002, 1 (2) :153-161.
[3] O.Fiehn . Combining genomics, metabolome analysis, and biochemical modeling to understand metabolic networks [J] .Comp Funct Genomics ,2001, (2): 155-168.
[4] J.K.Nicholson,I.D.Wilson. Opinion:understanding‘global’systems biology:Mera-bonomies and the continuum of metabolism.Nat Rev Drug Discov,2003,2:668-676.
[5] B.R.Baggett, J.D.Cooper , E.T. Hogan, et al . Profiling is of lavonoids found in legume root extracts using capillary electrophoresis[J]. Electrophoresis, 2002, 23 (11) : 1642-1651.
[6] J.L .Griffin , H.J.Williams ,et al. Metabolic profiling of genetic disorders : a multitissue 1H nuclear magnetic resonance spectroscopic and pattern recognition study into dystrophic tissue[J] . Anal Bio chem ,2001 ,293 (1) :16-21.
[7] Pelander,Ojanperal I,Lakes S.Toxicological screening with formula-based metabolite identification by liquid chromatography/ time-of-flight mass spectrometry [J] . Anal Chem ,2003 ,75(2):5710 -5718.
[8] Peiyuan Yina, Patamu Mohemaitib,et al.Serum metabolic profiling of abnormal savda by liquid-chromatography/mass-spectrometry[J].Journal of Chromatography B,2008 ,871(2):322-327.
[9] Bethanne, Warracka, et al. Normalization strategies for metabonomic analysis of urine samples[J]. Journal of Chromatography B, 2009,877(3): 547-552.
[10] Masahiro Sugimoto, David T. Wong, Akiyoshi Hirayama, etal.Capillary electrophoresis mass spectrometry-based saliva metabolomics identified oral, breast and pancreatic cancer-specific Profiles[J]. Metabolomics, 2010,6(1): 78-95.
[11] Kitano H. Systems biology: a brief overview[J]. Science, 2002,295 (5560) : 1662-1664.
[12] J.K.Nicholson, J.C.Lindon,HolmesE. Metabonomics: understanding the Metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data[J]. Xenobiotica.1999;29(11):1181-1189.
[13] FiehnO. Combining genomics, metabolome analysis, and biochemical modeling understand metabolic networks [J] .Comp Funct Genomics ,2001,2 :155-168.
[14]朱勇飞,张天宝.代谢组学在毒理学研究中的应用[M].国外医学:卫生学分册,2005 ,32(3) :156 - 159.
[15]杨军,宋硕林,Jose CP等.代谢组学及其应用[J].生物工程学报,2005,1(21):1-5.
[16]许国旺,杨军.代谢组学及其研究进展[J].色谱,2003,1(21): 316-320.
[17] C.W.Mun, J.Y.Cho, W.J.Shin.et a1.Exvivo proton MR spectroscopy(1H-MRS) for evaluation of human gastric carcinoma[J].Magn Reson Imaging, 2004, 22(6):861-870.
[18] Katz-Brull R, Seger.D, Rivenson-Segal.D, et a1. Metabolic markers of Breast cancer enhanced choline metabolism and reduced choline-ether-phospholipid synthesis[J].Cancer Res, 2002, 62(7) :1966-1970.
[19] Heintz P, Ehreneim Ch, Hundeshagen H. In Application of NMR Techniques on the Body Composition of Live Animals[EB/OL]. London, Elsevier, 1989 .
[20] Clare A, Daykin, Peta J.D. Foxall, Susan C.Connor, et al.The Comparison of Plasma Deproteinization Methods for the Detection of Low-Molecular-Weight Metabolites by 1H Nuclear Magnetic Resonance Spectroscopy[J]. Analytical Biochemistry ,2002,304(7):220-230.
[21] J. K. Nicholson , J. D . Foxall. 750 MHz IH and IH-l3C NMR Spectroscopy of Human Blood Plasma[J]. Anal.Chem, 1996,67(5):793-811.
[22] Kunle Odunsi,Robert M. Wollman,Christine B. Ambroson, Detection of epithelial ovarian cancer using 1H-NMR-based metabonomics[J], Int. J. Cancer, 2005,113(5):782-788.
[23]艾斯克尔.吐拉洪,哈木拉提.吾甫尔,豪富华.基于NMR的维吾尔医异常黑胆质型肿瘤患者血浆代谢组学分析[J],科技导报,2009,27(13):27-31.
[24]陆强,黄一红,丛辉等.肝细胞癌、肝硬化患者血清中代谢物组研究[J],分析化学,2009,39(2): 194-198.
[25]姜玉章,孙晓阳,史昆波,食管鳞癌患者血清1H NMR指纹图谱研究[J].南京医科大学学报,2009,29(11):1560-1563.
[26] H.J.Roeijmans, J.D.De Hoog, C.S.Tan. et al. Molecular taxonomy and GC/MS of metabolites of Scytalidum hyalinum and Nattrassia mangiferae (Hendersonula tofllloidea)[J], Med Vet Mycol, 1997, 35(3): 181-189.
[27] D.V.Huhman, L.W.Sumner. Metabolioc profiling of saponin glycosides in Medicago sativa and Medicago truncatula using HPLC coupled to an electrospray ion trap mass spectometer [J]. Phytochemistry, 2002, 59 (3) : 347-360.
[28] Soga T, Ueno Y, Naraoka H, et al.Simultaneous determination of anionic intermediates for bacillus subtilis metabolic pathways by capillary electrophoresis electros pray ionization mass spectrometry[ J ]. Anal Chem, 2002, 74 (10) : 2233-2239.
[29] H.J.Major, R.Williams, A.J.Wilson, I.D.Wilson. A metabonomic analysis of plasma from Zucker rat strains using gas chromatography/mass spectrometry and pattem recognition[J]. Rapid Commun Mass Spectrom ,2006,20(22): 3295-3302.
[30] D.S.Wishart, A.C.Guo, et al. HMDB: a knowledgebase for the human metabolome[DB]. Nucleic Acids Res. 2009 Jan; 37(Database issue): D603-610.
[31] J. T .Brindle, J. K. Nicholson, P. M.Schofield,et al. Application of chemometrics to 1H NMR spectroscopic data to investigate a relationship between human serum metabolic profiles and hypertension[J]. Analyst, 2003, 128(1):32-36.
[32] J.T..Brindle, H.Anni, E.Holmes, G.Tranter, J.K.Nicholson, H.W. Bethell, S .Clarke, P. M.Schofield, E. McKilligin, D. E. Mosedale, D. J. Grainger, Nat. Med. 2002, 8:1439-1442. [33 ] Ramadan Z, Jacobs D, Grigorov M, Kochhar S: Metabolic profiling using principal component analysis, discriminant partial least squares, and genetic algorithms[J]. Talanta, 2006, 68(5):1683-1691
[34] Odunsi K, Wollman RM, Ambrosone CB, Hutson A, McCann SE, Tammela J, Geisler JP, Miller G, Sellers T, Cliby W, Qian F, Keitz B, Intengan M, Lele S, Alderfer JL. Detection epithelial ovarian cancer using 1H-NMR-based metabonomics[J]. Int J Cancer, 2005, 113(5): 782-788.
[35] B.M. Beckwith-Hall, J.T .Brindle, R.H .Barton,et al. Application of orthogonal signal correction to minimise the effects of physical and biological variation in high resolution 1H NMR spectra of biofluids[J]. Analyst. 2002, 127:1283-1288.
[36] Holmes E,Antti H,Chemometric contributions to the evolution of metabonomics: mathematical solutions to characterising and interpreting complex biological NMR spectra[J]. Analyst, 2002, 127(12):1549-1557
[37] Millis K,Weybright P, CampbeilL N, et al . Classification of human liposarcoma and lipoma using exvivo proton NMR spectroscopy[J]. Magn Res on Med, 1999, 41 (2) : 257-267.
[38] J.T .Brindle,Antti H, Holmes E, et a1.Rapid and non invasive diagnosis of the presence and severity of coronary heart disease using 1H-NMR-based metabonomics[J]. Nat Med,2002,8(12):1439-1444.
[39] K.P.Chiang, Niessen S, Saghatelian A, et al. An enzyme that regulates ether lipid signaling pathways in cancer annotated by multi-dimensional profiling[J]. Chem Biol, 2006, 13 (10) : 1041-1050.
[40] J.L .Griffin , R.A. Kauppinen. A metabolomics perspective of human brain tumours [J] . FEBSJ , 2007 , 274 ( 5 ) : 1132 -1139.
[41] C.L.Florian, N.E.Preece, K.K.Bhakoo. Characteristic metabolic profiles revealed by 1 HNMR spectroscopy for three types of human brain and nervous system tumours[J].NMR Biomed , 995, 8(6) : 253 - 264.
[42] Odunsi K, R.M.Wollman, Ambrosone. Detection of epithelial ovarian cancer using 1 H-NMR-based metabonomics[J] . Cancer ,2005 ,113 (5) :782-788.
[43] Yang J. Diagnosis of liver cancer using HPLC-based metabonomics avoiding false-positive result from hepatitis and hepatocirrhosis diseases [J]. Chromatogr B Analyt Technol Biomed Life Sci ,2004 ,813 (1-2) :59- 65.
[44] Yang J , Xu G, Zheng Y. Strategy for metahonomics researchbased on high-performan liquid chromatography and liquidchromatography coupled with tandem mass spectromet ry[J ] .Chromatogr A ,2005 ,1084 (1 -2) : 214 - 221.
[45]陆强,黄一红,丛辉等.肝细胞癌、肝硬化患者血清中代谢物组研究[J],分析化学, 2009, 39(2):194-198.
[46]吴逸明等.高效液相色谱法测定肺癌患者尿中假尿嘧啶核苷和肌苷量比值[J].郑州大学学报(医学版)2002,37(4):433-435.
[47] Yan SK, Wei BJ, Lin ZY, et al. A metabonomic approach to the diagnosis of oral squamous cell carcinoma,oral lichen planus and oral ieukoplakia [J]. Oral Oncology, 2008, 44(5): 477-483.
[48]姜玉章,孙晓阳,史昆波.食管鳞癌患者血清1H NMR指纹图谱研究[J].南京医科大学学报,2009,29(11):1560-1563.
[49] Kubom A,M M Meguid,D C Hitch. Amino acid profiles correlate diagnostically with organ site in three kinds of malignant tumors[J]. Cancer, 1992,69(9): 2343-2348.
[50]汪晋,郁兰芳,沈朋,王书芳.气相色谱-质谱法分析乳腺癌患者血清代谢组[J],浙江大学报, 2009, 38(5): 45-48.
[51]何永频,陆瑞芳,吴岷.血清游离多不饱和脂肪酸的组分变化与乳腺癌的关系[J],中国临床营养杂志, 1999, 6(1): 21-23.
[52] Xu G, H.R.Schmid, ,et al. Excretionpattern investiation ofurinary normal and modified nucleosides of breas tcance rpatients by RP--HPLC andfaetoranalysis method[J]. Biomed Chromatogr, 2000, 14(7):459-466.
[53] Max Parkin,Freddie Bray,J.Ferlay,etal,Global Cancer Statistics,2002.CA Cancer ,2005,55:74-108
[54]中国乳腺癌流行病学调研项目[OL]: http://www.qgjk.org/html/2368.html.2011-03-25.
[55]李树云.触诊在乳癌诊断中的作用[J].江西医学院学报, 2002, 12( 5) : 48.
[56]于泽平,李幼生,王少华等.吸针穿刺吸取细胞学检查对乳腺癌的诊断价值[J].医学研究生学报, 2006,19(5) : 474-475.
[57]陈利平,乳腺癌发病机理的认识与治疗.2007亚太国际肿瘤生物学和医学学术会议暨首届解放军总医院肿瘤综合防治高峰论坛及第二届中国中青年肿瘤专家论坛论文集[C],2007.
[58]杨举伦,王丽.乳腺癌的分子病理学研究[J].临床与实验病理学杂志, 2011, 27(1): 10-14.
[59] Fattaneh A, T avassoli P D. Pathology and Genetics of the Breast and Female Genital Organs[ M ]. Lyon :I ARC Press , 2003: 49 -50.
[60]《乳腺癌HER2检测指南(2009版)》编写组.乳腺癌HER2检测指南( 2009版) [J].中华病理学杂志, 2009 , 38( 12) : 836-840.
[61]杨举伦,普苹,蔡学敏等.乳腺不典型增生组织中Hras基因突变的检测[J].临床与实验病理学杂志, 2001 , 17 ( 1 ) : 12-14.
[62]曹红婷,核磁共振代谢组学方法的体液制谱方法和制谱参数研究[D].厦门:厦门大学,2009:25-35.
[63] GoelA, J.R.anknecht. Concerted activation of ETS protein ER81 by p160 coactivators, the acetyltransferase p300 and the receptor tyrosine kinase HER2 /Neu[ J]. J Biol Chem, 2004, 279 ( 15) : 14909-14916.
[64]杨举伦,蔡学敏,普苹等.乳腺增生病P53基因第5外显子突变及其蛋白表达[J] .临床与实验病理学杂志, 2002 , 18( 3 ): 264-267.
[65]杨举伦, David K, Michael J B等.乳腺癌发生过程中NOEY2基因启动子区甲基化及mRNA表达[J] .临床与实验病理学杂志, 2005 , 21( 5 ) : 543-547.
[66] Yang J L , Ren Y, Wang L , et al . PTEN mutation spectrumin breast cancers and breast hyplasia[J]. J Cancer Res Clin Onco, 2010, 136( 9 ) : 1303-1311.
[67]杨海捷,王丽,魏万里等.散发性乳腺癌及乳腺不典型导管增生组织BRCA1基因启动子区甲基化分析[J] ,临床与实验病理学杂志, 2008 , 24( 2 ): 141- 145.
[68]谷永强,朱秋娟,肖俊英等. CA15-3和CEA检测在乳腺癌诊断中的意义[J],中国误诊学杂志, 2001, 1(11): 1619-1620.
[69]于鹏,杨兴无,杨春明等.乳腺癌手术前后肿瘤标志物CA153的检测及其临床意义[J],中华普通外科杂志, 2002, 17(8): 496.
[70]谢翠华,陆亚平. CA153在乳腺癌诊断和肿瘤分期中的应用[J],实用心脑肺血管病杂志, 2008, 16(11): 44-45.
[71]陈智周,范振符,杨剑等.肿瘤标志物CA153的免疫放射分析及其临床应用[J],中华肿瘤杂志, 1998, 20(2): 125-128..
[72] W.D.Dupont, F.F.Parl, W.H.Hartmann, et al. Breast cancer risk associated with proliferative breast disease and atypical hyperplasia[J]. Cancer, 1993, 71(4): 1258-1265.
[73]王鸿利.实验诊断学[M[.北京:人民卫生出版社, 2001: 240-242.
[74]朱海龙. CEA、AFP、CA125、CA15-3、CA19-9检测对恶性肿瘤的诊断价值探讨[J],现代医药卫生, 2006, 22(22): 3422-3423.
[75]朱波,黄文成,黄玲莎.血清CA153、CEA、TSGF联合检测在128例乳腺癌中的临床应用[J],福建医药杂志, 2003, 25(4): 155-174.
[76]王旭东.血清VEGF、CA153及CEA联合检测对乳腺癌的诊断价值[J],中国医师进修杂志, 2007, 30(4): 28-31.
[77]叶蓓,柳光宇,陆劲松.常用的乳腺癌血清肿瘤标志物在早期诊断中的应用价值不高[J],《中国癌症杂志》, 2009, 19(10): 807-808.
[78]沈坤炜,胡震.乳腺肿块的诊断和鉴别诊断[J].中国实用外科杂志,2005, 25(2):160-162.
[79]王颀,连臻强.乳腺癌早期诊断的临床思路[J],中华乳腺病杂志(电子版),2010,4(4):357-360.
[80] B.D.Ross, GK,Radda, D.G.Gadian,et a1. Preliminary observations on the metabolic responsesto exercise in humans, using 31-phosphorus nuclear magnetic resonance[J]. Ciba Found Symp,1982 ,87:145-152.
[81] Suna,SalnIiIlen,A.Soininen,P.Laatikamell,et al. l HNMR metabonomics of plasma lipoprotein subclasses:elucidation of metabolic clusteriIlng by self-oranising maps[J]. NMR Biomed, 2007, 20(7): 658-672.
[82] Akira K, Masu S, Imachi M, et al. 1HNMR-based metabonomic analysis of urine from young spontaneously hypertensive rats[J]. J Pham and Biomed Anal, 2008, 46(3): 550-556.
[83] Shi C, Wu CQ, Cao AM, et al. NMR-spectroscopy-based metabonomic approach to the analysis of bay41-4109, a novel anti-HBV compound,induced hepatotoxicity in rats[J]. Toxicol. Lett, 2007, 173(3):161-167.
[85] ] Liao PQ,Wei L,Zhang XY,et al. Metabolic pofiling of serum f rom gadolinium chloride Treated rats by 1HNMR spectroscopy[J]. Anal Biochem, 2007, 364(2):112-121.
[86] Ramadan Z, Jacobs D, et al. Metabolic Profiling using Principcal component diseriminant partial least squares,and genetic algoriths[J]. Talanta, 2006, 68(5):1683-1691.
[87] Liao PQ, Wei L, Zhang XY, et al. Metabolic profiling of serun from gadolinium chloride-treated Rats by 1HNMR,spectroscopy[J]. Anal Biotech, 2007, 364(2): 112-121.
[88] Wu HF, Zhang XY, Li XJ, et al. Comparison of metabolic profiles from serum from hepatotoxin-treted rats by nuclear-magnetic-resonnce-based metabonomic analysis [J]. Anal Biochem, 2005,340(1):99-105.
[89] Hewer R, Vorster, Steffens FE, et al. Applying biofluid 1HNMR-based metabonomic techniques to diatinguish between HIV-1 postive /AIDS patients on antiretroviral trentmentand HIV-1 negative individuala[J]. J Pham and Biomed Anal, 2006, 4(4): 1442-1446.
[90] J.T.Brindle, H.Anni, E.Holmes, J.K.Nicholson, H.W.Bethell, S.Clarke, P.M.Schofield, E .McKilligin, D.E. Mosedale, D. J. Grainger, Nat. Med. 2002, 8(11): 1439-1442.
[91] Ramadan Z, Jacobs D, Grigorov M, Kochhar S: Metabolic profiling using principal component analysis, discriminant partial least squares, and genetic algorithms[J]. Talanta ,2006, 68(5):1683-1691
[92] Odunsi K, R.M .Wollman, C.B. Ambrosone, Hutson A, S.E. McCann, Tammela J, J.P Geisler, Miller G, Sellers T, Cliby W, Qian F, Keitz B, Intengan M, Lele S, J.L .Alderfer. Detection epithelial ovarian cancer using 1H-NMR-based metabonomics[J]. Int J Cancer, 2005, 113(5): 782-788.
[93] Peiyuan Yin, Patamu Mohemaiti, Jing Chen, et al. Serum metabolic profiling of AbnormalSavda by liquid chromatography/mass spectrometry[J]. Journal of Chromatography B, 2008,43.(5): 1692-1698.
[94]冒海蕾,徐晏,王斌等.正交信号校正在正常人血清1H NMR谱的代谢组分析中的滤噪作用评价[J].化学学报, 2007, 65(2): 152-158.
[95]杨永霞,杨生义,梁敏锋.基于核磁共振波谱的乙肝患者血清代谢组研究[J],第二军医大学学报, 2010,31(3):388-291.
[96]阿仙姑·哈斯木,巴吐尔.买买提明,伊力亚尔.夏合丁.不同模式识别方法分析食管癌患者血浆代谢组的磁共振氢谱[J],中华肿瘤杂志, 2010, 32(9):681-684.
[97]高岗,杨根金,娄子洋.肾虚证大鼠尿液的核磁共振谱代谢组学研究[J].第二军医大学学报, 2009, 30: 565 - 568.
[98]艾斯克尔.吐拉洪,哈木拉提.吾甫尔,豪富华.基于NMR的维吾尔医异常黑胆质型肿瘤患者血浆代谢组学分析[J],科技导报,2009,27(13):27-31.
[99] C.W. Mun,J.Y. Cho,W.J .Shin, et a1. Ex vivo proton MR spectroscopy(1H-MRS) forevaluation of human gastric carcinoma[J].Magn Reson Imaging, 2004, 22(6):861-870.
[100] K.W .Jordan, L.L. Cheng. NMR-based metabolomics approach to target biomarkers for human prostate cancer [J]. Expert Rev Proteomics , 2007 , 4 (3): 389 - 400.
[101] K .Odunsi, R.M. Wollman, C.B.Ambrosone, et a1. Detection of epithelial ovarian cancer using 1 H-NMR-based metabonomics[J]. Int J Cancer, 2005, 113(5):782-788.
[102] J. E. Entire, K.C.Kuo, M.E.Smith, et a1. Classfication of lung-cancer patients and controls by chromatography of modified nucleosides in serum[J]. Cancer Res, 1989, 49(15):1057.
[103] Odunsi K, R.M .Wollman, C.B. Ambrosone. Detection of epithelial ovarian cancer using 1 H-NMR-based metabonomics[J]. 1nt J Cancer, 2005, 113 (5): 782 - 788.
[104] Denkert C , Budczies J , Kind T. Mass Specromery-based metabolic profiling reveals different metabolite patterns in invasive ovarian carcinomas and ovarian borderline tumors[J] . Cancer Res ,2006 ,66 (15): 10795-10804.
[105] Bethanne M.Warracka, Serhiy Hnatyshyna, Karl-Heinz Ott, Normalization strategies for metabonomicanalysisof urine samples[J]. Journal of Chromatography B, 2009, 877:547-552.
[106] S. Bijlsma, L.Bobeldijk, E.R..Verheij, et al. Large-scale human metabolomics studies: A strategy for data (pre-) processing and validation[J]. Anal Chem, 2006, 78(2): 567-574.
[107] G .B .Mills, W.H.Moolenaar. The emerging role of lysophosphatidic acid in cancer[J]. Nat Rev Cancer, 2003, 3(8): 582-591.
[108] P.Y .Yin, X. J .Zhao, Q .R,Li , J .S. Wang,et al. Metabonomics study of intestinal fistulas based on ultraperformance liquid chromatography coupled with Q-TOF massspectrometry (UPLC/Q-TOF MS)[J]. J Proteome Res, 2006, 5(9): 2135-2143.
[109] J.C.Lindon, E.Holmes, J.K.Nicholson. Metahonomics: Systems biology in pharmaceutical research and development[J]. Curr Opin Mol Ther, 2004,6(3): 265-272.
[110] User's Guide to SIMCA-P, SIMCA-P+ version 12.0.
[111]林青认,超高液相-质谱联用对胃癌代谢组学研究[D].杭州:浙江大学, 2010: 8-10.
[112] S.D .Hey, G.M.Park, M.A. MacnurLatl, et al. Stimulalation of protein synthesis in human tumors by parentetal nutrition:evidence for modulation of tumor growth[J]. Br J Surg, 1991, 78(4): 483-487.
[113]洪毅,谈冶雄,王红阳.代谢组学-肿瘤学研究的重要技术平台[J],中国肿瘤生物治疗杂志,2007,14(6):585-588.
[114] A.M .Proenza, Oliver J, Palou A, et al, Breast and lung cancer are associated with a decrease in blood cell aminoacid content[J]. J Nutr Biochem, 2003, 14(3):133-138.
[115]王喜,基于代谢组学的胃癌化疗敏感性和毒性预测[D].上海:第二军医大学, 2008:20-35.
[116] Cai H, Erhardt P. Hydrolysis of phosphatidylcholine couples Ras to activation of Raf protein kinase during mitogenic signal transduction[J]. Mol Cell Biol, 1993, 13(12): 7645-7651.
[117] Mulder C, L.O Wahlund, Decreased lysophosphatidylchoine/phosphatid ylcholine ratio in cerebrospinal fluid in Alzheimer's disease[J], J Neural Transm, 2003, 110(8): 949-955.
[118] Lee ES,Chen H,Inhibitory effects of lysophpsphatidylcholine on the dopaminergic system[J]. Neuro Chem Res, 2004, 29(7): 1333-1342.
[119] Y.L.Ting, D.Sherr. Variations in energy and Phospholipid metabolism in normal and cancer human mammary epithelial cells[J]. Anti caneer Res, 1996, 16(3B): 1381-1388.
[120] C.G.Gaetano, N.Samadi, Inhibition of autaxin prodution or activity blocks Lysophosphatidyl choline-induced migration of human breast cancer and mekanoma cells[J]. Mol Carcinog, 2009, 48(9): 801-809.
[121] D.N.Brindley, Hepatic secretion of lysphosphatidylcholine: A novel transport system for polyunsaturated fatty acids and choline[J]. J Nutr Biochem, 1993, 4: 442-449.
[122] E.Koh, R.W. Bandle, Novel point mutations attenuate autotaxin activity.Lipids [J]. Lipids Health Dis,2009,8:4
[123] Fang X, Schummer M, Lysophosphatidic acid is a bioactive meidtor in ovarian cancer[J], Biochim.biophys,actal, 2002, 82(15):254-264.
[124] Tarah L, Pua,Feng-qiang,Wang, Roles of LPA in ovarian cancer developmengt and proression[J], Future Oncol, 2009, 5(10):1649-1673