生理体液法分析肝病患者的氨基酸研究
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摘要
目的：(1)采用生理体液方法分析肝硬化、肝癌及慢性重型肝炎患者体液氨基酸的浓度,建立生理体液法分析氨基酸模型。(2)通过比较肝硬化、肝癌及慢性重型肝炎患者血清氨基酸代谢特点,探讨其氨基酸代谢模式。(3)通过分析肝硬化患者腹水氨基酸与血清氨基酸的变化,探讨其临床意义。
方法：随机选取天津市第三中心医院体检患者60例作为对照组(N组),选取住院患者70例,其中30例为肝硬化组(A组)、20例为肝癌组(B组)、20例为慢性重型肝炎组(C组),于入院后第一天早晨空腹抽取静脉血检测肝功能、凝血功能并采用生理体液法进行氨基酸分析。10例肝硬化伴腹水患者抽取腹水采用生理体液法进行氨基酸分析。
结果：(1)肝功能：与N组相比,试验组血清中白蛋白、前白蛋白浓度逐渐降低,血清谷草转氨酶、总胆汁酸、总胆红素浓度逐渐升高,C组最明显,与N组比较差异有统计学意义(P<0.05)。(2)凝血功能：与N组比较,试验组血清凝血酶原时间、活化部分凝血激酶时间及凝血酶时间逐渐延长,C组升高最明显,差异有统计学意义(P<0.05)；血清纤维蛋白原浓度降低,与N组比较,差异无显著统计学意义(P＞0.05)。(3)氨基酸分析结果：与N组比较,A组患者血清牛磺酸(Tau)、天门冬氨酸(Asp)、胱硫醚(Cysthi)、异亮氨酸(Ile)、精氨酸(Arg)浓度及支芳比值下降；p-丙氨酸(p-Ala)、p-氨基乙丁酸(p-Aiba)、γ-氨基乙丁酸(GABA)、α-氨基己二酸(α-AAA)、蛋氨酸(Met)、酪氨酸(Tyr)、苯丙氨酸(Phe)浓度升高,差异有统计学意义(P<0.05)。B组患者血清Asp、Cysthi、Ile、Arg、亮氨酸(Leu)、赖氨酸(Lys)浓度及支芳比值下降；Met、β-Ala、β-Aiba、GABA、α-AAA、Tyr、Phe、3甲基组氨酸(3Mehis)浓度升高,差异有统计学意义(P<0.05)。C组患者血清Tau、Asp、Cysthi、Ile、Leu、Lys、Arg、谷氨酸(Glu)、丙氨酸(Ala)、缬氨酸(Val)浓度及支芳比值下降；苏氨酸(Thr)、Met、β-Ala、β-Aiba、GABA、3Mehis、Tyr、Phe、1-甲基组氨酸(1Mehis)浓度升高,差异有统计学意义(P<0.05)。其中A组与B组比较,血清Val、Lys、Arg浓度下降,Tau、3Mehis浓度升高,差异有统计学意义(P<0.05)；A组与C组比较,血清Thr、Met、Tyr、3Mehis浓度升高,Ala、Glu、α-AAA、Val、Leu及Arg浓度下降,差异有统计学意义(P<0.05)；B组与C组比较,血清Thr、Met、Tyr浓度升高,Asp、Glu、Ala、Val、Cysthi及α-AAA浓度下降,差异有统计学意义(P<0.05)。(4)腹水氨基酸分析：与N组血清氨基酸比较,腹水Tau、Asp、Ser、Glu、Cysthi、Ile、Leu、α-氨基正丁酸(α-ABA)浓度降低,瓜氨酸(Cit)、胱氨酸(Cys)、et、Tyr、Phe、β-Ala、β-Aiba、GABA、3Mehis浓度升高,差异有统计学意义(P<0.05)。
结论：(1)成功采用生理体液法分析氨基酸,通过分析患者血清及腹水氨基酸的变化,得到更多氨基酸代谢信息,为研究肝脏疾病的氨基酸代谢变化提供了一种理想的方法；(2)通过比较肝硬化、肝癌及慢性重型肝炎患者与健康人血清氨基酸的变化,发现其氨基酸代谢变化不同,为临床诊治提供了一定的参考意义；(3)通过比较肝硬化、肝癌及慢性重型肝炎患者之间血清氨基酸浓度的差异,发现一些氨基酸在不同疾病进程中可能起重要作用,建立完善代谢物资源数据库,可能成为今后诊断和治疗肝病的努力方向；(4)与血清氨基酸相比较,腹水中氨基酸浓度变化与血清变化相一致,它同样反映了肝脏的损害情况,为揭示肝硬化的代谢异常提供了线索。
Objective:(1) we use physiological fluid method to analyze the concentration of amino acids in patients of hepatic ciirrhosis, hepatocellular carcinoma and chronic hepatitis and found a standard model of physiological fluid analysis. (2) By comparing different serum amino acids concentration of hepatic ciirrhosis, hepatocellular carcinoma and chronic hepatitis,we investigate the model of amino acids metabolism of different liver disease. (3)By analyzing the amino acid changes of ascites in patients with cirrhosis,we evaluate their clinical significance.
Methods:We randomly selected Tianjin Third Central Hospital of 60 patients as a medical control group.Experiment group was dedived into hepatic cirrhosis group(A group),hepatic carcinoma group(B group) and chronic severe hepatitis group(C group). In the first morning after admission,the blood samples were collected to carry out the liver function,blood coagulation and we adopted the physiological fluid method to analyze the amino acids. The ascites of 10 cases in liver cirrhosis patients with ascites was analyzed by physiological fluid method.
Results:(1)The liver function:Compared with the N group, serum albumin(Alb)、prealbumin of experimental group decreased gradually, and the serum aspartate aminotransferase, total bilirubin and total bile acid gradually increased, C group was the most significant compared with the N group (P＜0.05). (2)The blood coagulation:With the N group, the serum in prothrombin time, activated partial thromboplastin time and thrombin time of experimental group gradually extended, and C group increased most significantly(P＜0.05); the serum fibrinogen was decreased, and comparing with the N group, the difference was not statistically significant (P＞0.05). (3)The amino acids:Compared with the N group, the serum concentration of taurine(Tau)、aspartic acid(Asp)、isoleucine(Ile)、arginine(Arg)、cystathionine(Cysthi) and BCAA/AAA ratio of A group decreased significantly (P＜0.05); while the concentration of P-Alanine(β-Ala)、β-Amino isobutyric acid(β-Aiba)、γ-Amino-n-butyric acid(GABA)、α-amino adipic acid(a-AAA)、methionine(Met)、tyrosine(Tyr) and phenylalanine(Phe) increased significantly (P＜0.05). We found that these amino acids(Asp、Cysthi、Ile、Arg、leucine(Leu)、 lysine(Lys))and BCAA/AAA ratio was decreased in B group(P＜0.05), and the concentration of Met、β-Ala、β-Aiba、GABA、α-AAA、Tyr、Phe、3-Methylhistidine(3Mehis) was higher than N group (P＜0.05). The concentration of Tau、Asp、Cysthi、ILe、Leu、Lys、Arg、glutamic acid(Glu)、alanine(Ala)、valine(Val) and BCAA/AAA ratio in C group was decreased(P＜0.05); while Met、β-Ala、β-Aiba、GABA、3Mehis、Tyr、Phe、threonine(Thr)、1-Methylhistidine(1Mehis) increased significantly(P＜0.05). Between A and B group, the concentration of Val、Lys and Arg decreased significantly (P＜0.05), Tau and 3Mehis was increased significantly (P＜0.05). While compared A and C group, serum concentration of Thr、Met、Tyr、3Mehis was increased significantly (P＜0.05), Ala、Glu、α-AAA、Val、Leu and Arg was decreased significantly(P＜0.05). While compared B and C group, serum concentration of Thr、Met、Tyr was increased significantly (P＜0.05), the concentration of Asp、Glu、Ala、Val、Cysthi and a-AAA was decreased significantly(P＜0.05). (4)The amino acids concentration in ascites:With the N group, the concentration of Tau、Asp、Ser、Glu、Cysthi、Ile、Leu、α-amino-n-butyric acid(α-ABA) was decreased, while the concentrition of citrulline(Cit)、cystine(Cys)、Met、Tyr、Phe、β-Ala、β-Aiba、GABA and 3Mehis was increased significantly(P＜0.05).
Conclusion:(1) We have a successful adoption of physiological fluid method in amino acid analysis, and by analyzing different amino acids in serum metabolites in various patients, we get more information about amino acids metabolism in petients. It provides an ideal method for the study of amino acid metabolism in liver disease. (2) By comparing amino acids concentration in serum of hepatic cirrhosis、hepatocellular carcinoma and chronic hepatitis patients with N group, we found they have different changes in amino acid metabolism and it provides some reference value for clinical diagnosis. (3) By comparing the difference between the serum amino acid concentrations in hepatic cirrhosis、hepatocellular carcinoma and chronic hepatitis, we found that a number of amino acids play an important role in the disease process, and establishing and improving metabolites resource database may be the direction in diagnosis and treatment of liver disease in the future. (4) Compared with the serum amino acids, the changesof amino acids in ascites and serum in hepatic ciirrhosis patients was consistent, it also reveals liver damage and provid the clues of metabolic abnormalities in hepatic cirrhosis.
方法：随机选取天津市第三中心医院体检患者60例作为对照组(N组),选取住院患者70例,其中30例为肝硬化组(A组)、20例为肝癌组(B组)、20例为慢性重型肝炎组(C组),于入院后第一天早晨空腹抽取静脉血检测肝功能、凝血功能并采用生理体液法进行氨基酸分析。10例肝硬化伴腹水患者抽取腹水采用生理体液法进行氨基酸分析。
结果：(1)肝功能：与N组相比,试验组血清中白蛋白、前白蛋白浓度逐渐降低,血清谷草转氨酶、总胆汁酸、总胆红素浓度逐渐升高,C组最明显,与N组比较差异有统计学意义(P<0.05)。(2)凝血功能：与N组比较,试验组血清凝血酶原时间、活化部分凝血激酶时间及凝血酶时间逐渐延长,C组升高最明显,差异有统计学意义(P<0.05)；血清纤维蛋白原浓度降低,与N组比较,差异无显著统计学意义(P＞0.05)。(3)氨基酸分析结果：与N组比较,A组患者血清牛磺酸(Tau)、天门冬氨酸(Asp)、胱硫醚(Cysthi)、异亮氨酸(Ile)、精氨酸(Arg)浓度及支芳比值下降；p-丙氨酸(p-Ala)、p-氨基乙丁酸(p-Aiba)、γ-氨基乙丁酸(GABA)、α-氨基己二酸(α-AAA)、蛋氨酸(Met)、酪氨酸(Tyr)、苯丙氨酸(Phe)浓度升高,差异有统计学意义(P<0.05)。B组患者血清Asp、Cysthi、Ile、Arg、亮氨酸(Leu)、赖氨酸(Lys)浓度及支芳比值下降；Met、β-Ala、β-Aiba、GABA、α-AAA、Tyr、Phe、3甲基组氨酸(3Mehis)浓度升高,差异有统计学意义(P<0.05)。C组患者血清Tau、Asp、Cysthi、Ile、Leu、Lys、Arg、谷氨酸(Glu)、丙氨酸(Ala)、缬氨酸(Val)浓度及支芳比值下降；苏氨酸(Thr)、Met、β-Ala、β-Aiba、GABA、3Mehis、Tyr、Phe、1-甲基组氨酸(1Mehis)浓度升高,差异有统计学意义(P<0.05)。其中A组与B组比较,血清Val、Lys、Arg浓度下降,Tau、3Mehis浓度升高,差异有统计学意义(P<0.05)；A组与C组比较,血清Thr、Met、Tyr、3Mehis浓度升高,Ala、Glu、α-AAA、Val、Leu及Arg浓度下降,差异有统计学意义(P<0.05)；B组与C组比较,血清Thr、Met、Tyr浓度升高,Asp、Glu、Ala、Val、Cysthi及α-AAA浓度下降,差异有统计学意义(P<0.05)。(4)腹水氨基酸分析：与N组血清氨基酸比较,腹水Tau、Asp、Ser、Glu、Cysthi、Ile、Leu、α-氨基正丁酸(α-ABA)浓度降低,瓜氨酸(Cit)、胱氨酸(Cys)、et、Tyr、Phe、β-Ala、β-Aiba、GABA、3Mehis浓度升高,差异有统计学意义(P<0.05)。
结论：(1)成功采用生理体液法分析氨基酸,通过分析患者血清及腹水氨基酸的变化,得到更多氨基酸代谢信息,为研究肝脏疾病的氨基酸代谢变化提供了一种理想的方法；(2)通过比较肝硬化、肝癌及慢性重型肝炎患者与健康人血清氨基酸的变化,发现其氨基酸代谢变化不同,为临床诊治提供了一定的参考意义；(3)通过比较肝硬化、肝癌及慢性重型肝炎患者之间血清氨基酸浓度的差异,发现一些氨基酸在不同疾病进程中可能起重要作用,建立完善代谢物资源数据库,可能成为今后诊断和治疗肝病的努力方向；(4)与血清氨基酸相比较,腹水中氨基酸浓度变化与血清变化相一致,它同样反映了肝脏的损害情况,为揭示肝硬化的代谢异常提供了线索。
Objective:(1) we use physiological fluid method to analyze the concentration of amino acids in patients of hepatic ciirrhosis, hepatocellular carcinoma and chronic hepatitis and found a standard model of physiological fluid analysis. (2) By comparing different serum amino acids concentration of hepatic ciirrhosis, hepatocellular carcinoma and chronic hepatitis,we investigate the model of amino acids metabolism of different liver disease. (3)By analyzing the amino acid changes of ascites in patients with cirrhosis,we evaluate their clinical significance.
Methods:We randomly selected Tianjin Third Central Hospital of 60 patients as a medical control group.Experiment group was dedived into hepatic cirrhosis group(A group),hepatic carcinoma group(B group) and chronic severe hepatitis group(C group). In the first morning after admission,the blood samples were collected to carry out the liver function,blood coagulation and we adopted the physiological fluid method to analyze the amino acids. The ascites of 10 cases in liver cirrhosis patients with ascites was analyzed by physiological fluid method.
Results:(1)The liver function:Compared with the N group, serum albumin(Alb)、prealbumin of experimental group decreased gradually, and the serum aspartate aminotransferase, total bilirubin and total bile acid gradually increased, C group was the most significant compared with the N group (P＜0.05). (2)The blood coagulation:With the N group, the serum in prothrombin time, activated partial thromboplastin time and thrombin time of experimental group gradually extended, and C group increased most significantly(P＜0.05); the serum fibrinogen was decreased, and comparing with the N group, the difference was not statistically significant (P＞0.05). (3)The amino acids:Compared with the N group, the serum concentration of taurine(Tau)、aspartic acid(Asp)、isoleucine(Ile)、arginine(Arg)、cystathionine(Cysthi) and BCAA/AAA ratio of A group decreased significantly (P＜0.05); while the concentration of P-Alanine(β-Ala)、β-Amino isobutyric acid(β-Aiba)、γ-Amino-n-butyric acid(GABA)、α-amino adipic acid(a-AAA)、methionine(Met)、tyrosine(Tyr) and phenylalanine(Phe) increased significantly (P＜0.05). We found that these amino acids(Asp、Cysthi、Ile、Arg、leucine(Leu)、 lysine(Lys))and BCAA/AAA ratio was decreased in B group(P＜0.05), and the concentration of Met、β-Ala、β-Aiba、GABA、α-AAA、Tyr、Phe、3-Methylhistidine(3Mehis) was higher than N group (P＜0.05). The concentration of Tau、Asp、Cysthi、ILe、Leu、Lys、Arg、glutamic acid(Glu)、alanine(Ala)、valine(Val) and BCAA/AAA ratio in C group was decreased(P＜0.05); while Met、β-Ala、β-Aiba、GABA、3Mehis、Tyr、Phe、threonine(Thr)、1-Methylhistidine(1Mehis) increased significantly(P＜0.05). Between A and B group, the concentration of Val、Lys and Arg decreased significantly (P＜0.05), Tau and 3Mehis was increased significantly (P＜0.05). While compared A and C group, serum concentration of Thr、Met、Tyr、3Mehis was increased significantly (P＜0.05), Ala、Glu、α-AAA、Val、Leu and Arg was decreased significantly(P＜0.05). While compared B and C group, serum concentration of Thr、Met、Tyr was increased significantly (P＜0.05), the concentration of Asp、Glu、Ala、Val、Cysthi and a-AAA was decreased significantly(P＜0.05). (4)The amino acids concentration in ascites:With the N group, the concentration of Tau、Asp、Ser、Glu、Cysthi、Ile、Leu、α-amino-n-butyric acid(α-ABA) was decreased, while the concentrition of citrulline(Cit)、cystine(Cys)、Met、Tyr、Phe、β-Ala、β-Aiba、GABA and 3Mehis was increased significantly(P＜0.05).
Conclusion:(1) We have a successful adoption of physiological fluid method in amino acid analysis, and by analyzing different amino acids in serum metabolites in various patients, we get more information about amino acids metabolism in petients. It provides an ideal method for the study of amino acid metabolism in liver disease. (2) By comparing amino acids concentration in serum of hepatic cirrhosis、hepatocellular carcinoma and chronic hepatitis patients with N group, we found they have different changes in amino acid metabolism and it provides some reference value for clinical diagnosis. (3) By comparing the difference between the serum amino acid concentrations in hepatic cirrhosis、hepatocellular carcinoma and chronic hepatitis, we found that a number of amino acids play an important role in the disease process, and establishing and improving metabolites resource database may be the direction in diagnosis and treatment of liver disease in the future. (4) Compared with the serum amino acids, the changesof amino acids in ascites and serum in hepatic ciirrhosis patients was consistent, it also reveals liver damage and provid the clues of metabolic abnormalities in hepatic cirrhosis.
引文
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[41]Wu H, Xue R, Dong L, et al. etabolic profiling of human urine in hepatocellular carcinoma patients using gas chromatography/mass spectrometry[J]. Anal Chim Acta,2009,648(1):98-104.
[42]Kimura T, Noguchi Y, Shikata N, et al. Plasma amino acid analysis for diagnosis and amino acid-based metabolic networks[J]. Curr Opin Clin Nutr Metab Care,2009,12(1):49-53.
[43]王景泉,姜太一,沈成利,等.血清氨基酸指标在病毒性肝炎及肝炎肝硬化诊断中的意义[J].临床肝胆病杂志,2003，19(3)：188-190.
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[1]Hirs HW, Moore S, Stein WH. The chromatography of amino acid on ion exchange resins use of volatile acids for elution[J]. Journal of American Chemical Society,1954,76(1):6063-6065.
[2]Moore S, Spackman DH, SteinWH. Chromatography of amino acids on sulfonated polystyrene resins[J]. Aanlytical Chemistry,1958,30(7):1185-1190.
[3]Spackman DH, Stein WH, Moore S. Automatic recording apparatus for use in the chromatography of amino acids[J]. Analytical Chemistry,1958, 30(7):1190-1206.
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[1]Hirs HW, Moore S, Stein WH. The chromatography of amino acid on ion exchange resins use of volatile acids for elution[J]. Journal of American Chemical Society,1954,76(1):6063-6065.
[2]Moore S, Spackman DH, SteinWH. Chromatography of amino acids on sulfonated polystyrene resins[J]. Aanlytical Chemistry,1958,30(7):1185-1190.
[3]Spackman DH, Stein WH, Moore S. Automatic recording apparatus for use in the chromatography of amino acids[J]. Analytical Chemistry,1958, 30(7):1190-1206.
[4]Kirk AD, Frederick LA, Glover SG. Optical Resolution of the D-and L-amino acid family by liquid-solid chromatography[J]. Journal of American Chemical Society,1980,102:7122-7123.
[5]Dobashi A, Hara S. Optical resolution of the D-and L-amino acid derivatives by silica gel liquid chromatohy with chiral additive N-acetyl-L-valine tert-butylamide[J]. Anaiytical Chemistry,1983,55:1805-1806.
[6]Nolan TG, Hart BK, Doviehi NJ. Photothermal refraction as a microbore liquid chromatography detector in femtomde amino acid determination[J]. Analytical Chemistry,1985,57:2703-2705.
[7]Gartenmann K, Kochhar S. Short chain peptide analysis by high performance liquid chromatography coupled to electrospray ionization mass spectrometer after derivatization with 9-fluorenylmethyl chloroformate[J]. J Agric Food Chem,1999,47(12):5068-5071.
[8]Frank MP, Powers RW. Simple and rapid quantitative high-performance liquid chromatographic analysis of plasma amino acids[J]. J Chromatogr B,2007, 852(1/2):646-649.
[9]Krizmn M, Virant K, Proek M. Determination of derivatized amino acids in human embryo culture media by gas chromatography[J]. J Chromatogr B,2007, 858(1/2):292-295.
[10]Tuma P, Samcov AE, Andelova K. Determination of free amino acids and related compounds in amniotic fluid by capillary electrophoresis with contactless conductivity detection[J]. J Chromatogr B,2006,839(1/2):12-18.
[11]朱曙东,赵异皓.氨基酸的高效液相色谱分析[J].色谱,1994,12(1)：20-24.
[12]Rfeifer R, Karol R, Burgoyne R, et al. Application of HPLC to amino acid analysis[J]. American Laboratery,1983,15(3):77-78.
[13]刘惠文.柱前和柱后衍生高效液相色谱分析[J].氨基酸方法进展与评述,1995,17(2)：50-55.
[14]Kopple JD. Effect of nutrition on morbidity and mortality in maintenance dialysis patients[J]. Am J Kidney Dis,1994,24(6):1002-1009.
[15]吴华,徐中武,唐志毅.肾病综合征患者氨基酸代谢临床观察[J].临床荟萃,2003,18(10)：563-564.
[16]Holm Eggrrt, Sedlaczed Oliver, Grips Eva. Amino Acid metabolism in liver disease[J]. Curr Opin Clin Nutr Metab Care,1999,2(1):47-53.
[17]Kano T, Nagaki M, Takahashi T, et al. Plasma free amino acid pattern in chronic hepatitis as sensitive and prognostic index[J]. Gastroenterol JPN,1991, 26(3):344-349.
[18]杨芊,李兰娟,傅素珍.慢性重型肝炎患者氨基酸研究[J].浙江医学,2000,22(10)：586-588.
[19]杨晶,陆伟,邹满意等.荷肝癌小鼠血浆和肿瘤组织游离氨基酸代谢变化及其与肿瘤体积相关性研究[J].实用肝脏病杂志,2006,9(1)：22-24.
[20]Bozzetti F, Gavazzi C, Cozzaglio L, et al. Total Parenteral nutrition and tumor growth in malnourished patients gastric cancer[J]. Tumori,1999, 85(3):163-166.
[21]Nishihira T, Takagi T, Mori S. Leucine and manifestation of antitumor activity by valine-depleted amino acid imbalance[J]. Nutrition,1993,9(2):146-152.
[22]Fu YM, Yu ZX, Fe VJ, et al. Tyrosine and phenylalanin restriction induces G0/G1 cell cycle arrest in murine mclanoma in vitro and in vivo[J]. Nutr Cancer, 1997,29(2):104-113.
[23]Hoshiya Y, Guo H, Kubota T, et al. Human tumors are methionine dependent in vivo[J].Anticancer Res,1995,15(3):717-718.
[24]Parnetti L. Hyperhomocysteinemia:a risk factor for cerebrovascular disease[J]. Clin Exp Hypertents,2002,24(7):501-510.
[25]Wald DS, Law M, Morris JK. Homocysteine and cardiovascular disease:evidence on causality from a meta-analysis[J]. B M J,2002, 325(7374):1202-1225.
[26]Tsukishiro T, Shimizu Y, Higuchi K, et al. Efect of branched-chain amino acids on the composition cytolytic activity of liver-associated lymphocytes in rats[J]. J Gastroenterol Hepatol,2002,15(8):849-859.
[27]Als-Nielsen B, Koretz RL, Kjaergard LL, et al. Branched-chain amino acids for hepatic encephalopathy(Cochrane review). In:The Cochrane Library Issue 2. Chichester:John Wiley, Sons, Ltd,2003.
[28]The Italian BCAA Study Group. Nutritional supplementation with branched chain amino acids in advanced cirhosis:a double blind, randomized trial[J]. Gastroenterology,2003,124(7):1792-1801.
[29]Muto Y, Sato S, Watanabe A, et al. Effects of oral branched chain amino acid granules on event-free survival in patients with liver cirhosis[J]. Clin Gastroenterol Hepatol,2005,3(7):705-713.
[30]Saito Y, Saito H, Nakamura M, et al. Effect of the molar ratio of branched-chain to aromatic amino acids on growth and albumin mRNA expression of human liver cancer cell lines in a serum-free medium [J]. Nutr Cancer,2001, 39(1):126-131.
[31]Cerra FB, Upson D, Angelico R, et al. Branched-chains spport postoperative protein synthesis[J]. Surgery,1982,92(2):192-199.
[32]Garcia-de-Lorenzo A, Ortiz-Leyba C, Planas M, et al. Parenteral administration of diferent amounts of branch-chain amino acids in septic patients:clinical and metabolic aspects[J]. Crit Care Med,1997,25(3):418-424.
[33]Meric F, Buchholz TA, Mirza NQ, et al. Long-term complications associated with breast-conservation surgery and radio therapy[J].Ann Surg Oncol,2002, 9(6):543-549.
[34]Kokkinakis DM, Hoffman RM, Frenkel EP, et al. Synergy between methionine stress and chemotherapy in the treatment of brain tumor xenografts in athymic mice[J]. Cancer Res,2001,61 (10):4017-4023.
[35]Van Bokhorst DC, Van Der Schueren MA, Quak JJ, et al. Effect of perioperative nutrition with and without arginine supplementation on nutrirtional status, immune function, postoperative morbidity and survival in severely malnourished head and neck cancer patients[J]. Am J Clin Nutr,2001, 73(2):323-332.