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浙江大学学报（医学版） 2012年第2期41卷 188-191页

作者：刘二刚傅旭春钱莺白海波浙江大学城市学院药物研究所浙江杭州310015 浙江大学药学院浙江杭州310058 浙江大学医学院附属妇产科医院浙江杭州310006 杭州华东医药集团生物工程研究所有限公司浙江杭州310012

目的:建立一种测定大鼠肾组织中羟脯氨酸(Hyp)含量的柱前衍生HPLC法。方法:将大鼠肾组织以6 mol·L-1盐酸于110℃水解24 h,加入9-芴甲氧基羰酰氯(FMOC)衍生后,进行HPLC分析。HPLC分析条件为:Phenomenex C18色谱柱(4.60 mm×150 ... 详细信息

目的:建立一种测定大鼠肾组织中羟脯氨酸(Hyp)含量的柱前衍生HPLC法。方法:将大鼠肾组织以6 mol·L-1盐酸于110℃水解24 h,加入9-芴甲氧基羰酰氯(FMOC)衍生后,进行HPLC分析。HPLC分析条件为:Phenomenex C18色谱柱(4.60 mm×150 mm,5(m);流动相:醋酸钠缓冲液(pH3.78)-甲醇-乙腈(梯度洗脱);流速:1.0 ml·min-1;柱温:40℃;检测波长:265 nm。结果:羟脯氨酸在浓度范围为15.30～612.00 mg.L-1内线性良好(r=0.9999);回收率为97.4%～103.9%。结论:该方法简单、准确、灵敏,可用于测定大鼠肾组织中羟脯氨酸的含量。

关键词：色谱法,高压液相肾/代谢羟脯氨酸/分析离心法,梯密度柱前衍生肾组织

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兔耳增生性瘢痕的形态学观察与羟脯氨酸含量变化

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中国组织工程研究 2002年第8期6卷 1226-1227页

作者：舒彬郝林林吴宗耀黄显凯沈岳袁纯唐其敏 Daping Hospital Third Military Medical University Chongqing 400042China

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正常家兔角膜羟脯氨酸与氨基己糖的测定方法

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中华眼科杂志 1986年第2期22卷 104-107页

作者：应世灏天津医学院附属医院眼科教研室

本文报告一种简便的、准确的测定角膜羟脯氨酸和氨基己糖的方法。以6N HCl、7 h、100℃的条件水解角膜标本(2～5 mg)。基本采用 Stagman 的方法测定羟脯氨酸,但将脱醛乙醇代替乙二醇甲醚,对二甲氨基苯甲醛的浓度由20%改为8%(W/V)。基本... 详细信息

本文报告一种简便的、准确的测定角膜羟脯氨酸和氨基己糖的方法。以6N HCl、7 h、100℃的条件水解角膜标本(2～5 mg)。基本采用 Stagman 的方法测定羟脯氨酸,但将脱醛乙醇代替乙二醇甲醚,对二甲氨基苯甲醛的浓度由20%改为8%(W/V)。基本采用 Elson 方法测定氨基己糖,但乙酰化在密封安瓿内、pH 9.5,1 h、92±2℃进行。测定前采用 Zerolit 离子交换树脂柱提纯,除去干扰物质。测得家兔角膜羟脯氨酸和盐酸氨基己糖的含量各为8.53±0.26%(W/W)和1.96±0.13%(W/W)。

关键词：己糖胺类/分析角膜/化学羟脯氨酸/分析

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化疗物对切口组织中羟脯氨酸含量的影响

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实用肿瘤学杂志 1994年第2期8卷 11页

作者：张艳徐广伟张洋齐红刘玉霞董永兰张云五张强程龙伟张卓娄刚毅许亚明吉林省肿瘤医院

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DIFFERENTIAL PULSE POLAROGRAPHIC-DETERMINATION OF SOME CARCINOGENIC NITROSAMINES

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ANALYTICAL CHEMISTRY 1975年第14期47卷 2412-2418页

作者： HASEBE, K OSTERYOUNG, J COLORADO STATE UNIV DEPT CIVIL ENGFT COLLINSCO 80523 COLORADO STATE UNIV DEPT MICROBIOLFT COLLINSCO 80523

The pulse polarographic behavior of N-nitrosopyrrolidine and two of its derivatives, N-nitrosoproline and N-nitroso-4-hydroxyproline, has been investigated in buffered solutions over a range of pH. The effects of temperature, mercury pressure, modulation amplitude, surfactant concentration, and solvent and supporting electrolyte have been studied. The reduction process is complicated and pH-dependent. In acid solutions, N-nitrosopyrrolidine and its derivatives give one irreversible reduction wave, while for pH greater than 6, only N-nitrosopyrrolidine itself is reduced. The reduction waves of N-nitrosoproline and N-nitroso-4-hydroxyproline at pH below 5 are diffusion-controlled in dc and normal pulse polarographic modes. In aqueous medium, N-nitrosopyrrolidine, N-nitrosoproline, and N-nitroso-4-hydroxyproline have been detected quantitatively at the 10^-7 M level by differential pulse polarography with adequate precision and accuracy. The importance of examining electrochemical behavior to aid in selecting conditions for analysis is discussed. © 1975, American Chemical Society. All rights reserved.

关键词：致癌物/分析羟脯氨酸/类似物和衍生物羟脯氨酸/分析亚硝胺类/分析极谱法/方法脯氨酸/类似物和衍生物脯氨酸/分析吡咯烷类/分析溶剂温度

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Functional tissue engineering of articular cartilage through dynamic loading of chondrocyte-seeded agarose gels

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JOURNAL OF BIOMECHANICAL ENGINEERING-TRANSACTIONS OF THE ASME 2000年第3期122卷 252-260页

作者： Mauck, RL Soltz, MA Wang, CCB Wong, DD Chao, PHG Valhmu, WB Hung, CT Ateshian, GA Columbia Univ Dept Biomed Engn Cellular Engn Lab New York NY 10027 USA Columbia Univ Dept Mech Engn New York NY 10027 USA Columbia Univ Dept Orthopaed Surg Orthopaed Res Lab New York NY 10032 USA

Due to its avascular nature, articular cartilage exhibits a very limited capacity to regenerate and to repair. Although much of the tissue-engineered cartilage in existence has been successful in mimicking the morphological and biochemical appearance of hyaline cartilage, it is generally mechanically inferior to the natural tissue. In this study, we tested the hypothesis that the application of dynamic deformational loading at physiological strain levels enhances chondrocyte matrix elaboration in cell-seeded agarose scaffolds to produce a more functional engineered tissue construct than in free swelling controls. A custom-designed bioreactor was used to load cell-seeded agarose disks dynamically in unfolded compression with a peak-to-peak compressive strain amplitude of 10 percent, at a frequency of 1 Hz, 3 X (1 hour on, 1 hour off)/day, 5 days/week for 4 weeks. Results demonstrated that dynamically loaded disks yielded a sixfold increase in the equilibrium aggregate modulus over free swelling controls after 28 days of loading (100 +/- 16 kPa versus 15 +/- 8 kPa, p<0.001). This represented a 21-fold increase over the equilibrium modulus of day 0 (4.8+/-2.3 kPa). Sulfated glycosaminoglycan content was also found to be greater in dynamically loaded disks compared to free swelling controls at day 21 (p<0.0001 and p=0.002, respectively).

关键词：软骨关节/细胞学软骨关节/生长和发育软骨细胞/化学软骨细胞/生理学抗压强度培养技术/方法凝胶类葡糖氨基聚糖类/分析羟脯氨酸/分析琼脂糖应力物理时间因素负重动物牛

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SIMPLE PROCEDURES FOR DETERMINATION OF [C-14]HYDROXYPROLINE

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ANALYTICAL BIOCHEMISTRY 1975年第2期66卷 330-339页

作者： BLUMENKRANTZ, N ASBOEHANSEN, G UNIV COPENHAGEN RIGS HOSPDEPT DERMATOLCONNECT TISSUE LAB2100 COPENHAGENDENMARK

Two improved and simplified procedures are presented for determination of [ 14 C]hydroxyproline. Simplification is achieved by boiling samples without previous toluene extractions and, after boiling, passing the toluene extracts through a silicic acid column. Procedure I avoids incomplete toluene extraction of [ 14 C]proline derivatives and uses instead a specific adsorption to a silicic acid column. Procedure IIA introduces inter alia the silicic acid column to reduce the interference of incompletely extracted [ 14 C]proline. Procedure IIB simplifies procedure IIA by replacing extraction of [ 14 C]proline derivatives with a silicic acid column. The new procedures are simple, handy, specific and reproducible methods for determination of [ 14 C]hydroxyproline and are preferable to any other method known today. Procedure IIB is specially recommended for routine use.

关键词：碳放射性同位素评价研究羟脯氨酸/分析指示剂和试剂方法脯氨酸

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BREAKDOWN OF PROTEOGLYCAN AND COLLAGEN INDUCED IN PIG ARTICULAR-CARTILAGE IN ORGAN-CULTURE

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ANNALS OF THE RHEUMATIC DISEASES 1975年第4期34卷 303-311页

作者： DINGLE, JT HORSFIELD, P FELL, HB BARRATT, MEJ STRANGEWAYS RES LAB TISSUE PHYSIOL DEPTWORTS CAUSEWAYCAMBRIDGE CB1 4RNENGLAND UNIV CAMBRIDGE DEPT PATHOLIMMUNOL DIVCAMBRIDGEENGLAND

Explants of articular cartilage from young pigs were maintained in organ culture for 10--16 days, and degradation of matrix was induced by retinol or complement-sufficient antiserum. The percentage breakdown of proteoglycan and collagen (as hydroxyproline release) was measured. The response of the cartilage depended on whether or not the explants were cut so as to include some of the invading marrow ('invasion zone'). In media containing retinol, cartilage lost up to three-quarters of its proteoglycan whether the invasion zone was present or not, but very little of its collagen unless this region was included. In the presence of complement-sufficient anti-serum, however, cartilage without the invasion zone was virtually unaffected, but both proteoglycan and hydroxyproline were released when invasion zone was included; here proteoglycan release began almost immediately, but there was a time-lag of 6--8 days before a substantial amount of hydroxyproline appeared in the medium. Histological examination of sample explants from the experiments supported the biochemical findings. The possible significance of the results in relation to rheumatoid arthritis is discussed.

关键词：骨髓/分析骨髓/代谢软骨关节/分析软骨关节/药物作用软骨关节/代谢胶原/分析胶原/代谢补体系统蛋白质类葡糖氨基聚糖类/代谢羟脯氨酸/分析免疫血清/药理学器官培养技术蛋白聚糖类/分析蛋白聚糖类/代谢猪维生素A/药理学动物

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ISOLATION AND INITIAL CHARACTERIZATION OF HUMAN BASEMENT-MEMBRANE COLLAGENS

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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 1977年第2期76卷 376-384页

作者： TRELSTAD, RL LAWLEY, KR HARVARD UNIV MASSACHUSETTS GEN HOSPSCH MED SHRINERS BURNS INSTDEPT PATHOLEXPTL PATHOL LAB BOSTON MA 02114 USA

Neutral solutions of pepsin extracted human collagens derived from glomeruli, kidney, aorta, lung, heart, bowel, spleen, skeletal muscle and skin were subjected to heat gelation at ***. C. Centrifugation of the gel provided 2 fractions: gelled pellet and non-gelled supernatant. Analysis of these 2 fractions by gel electrophoresis, molecular sieve and ion exchange chromatography, and amino acid and carbohydrate determinations indicated that the non-gelled supernatant contained a substantial enrichment of basement membrane like collagen. The initial characterization of lung derived basement membrane collagen indicated close similarities with those derived from glomeruli and whole kidney and differences with that obtained from the spleen.

关键词：氨基酸类/分析基膜/分析胶原/分析胶原/分离和提纯热温度羟赖氨酸/分析羟脯氨酸/分析肾肾小球肺脾人类

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An oral endothelin-A receptor antagonist blocks collagen synthesis and deposition in advanced rat liver fibrosis

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GASTROENTEROLOGY 2000年第6期118卷 1169-1178页

作者： Cho, JJ Hocher, B Herbst, H Jia, JD Ruehl, M Hahn, EG Riecken, EO Schuppan, D Univ Erlangen Nurnberg Dept Med 1 D-91054 Erlangen Germany Free Univ Berlin Klinikum Benjamin Franklin Dept Gastroenterol D-12200 Berlin Germany Humboldt Univ Dept Nephrol Berlin Germany Univ Hosp Eppendorf Inst Pathol Hamburg Germany

Background & Aims: Endothelin 1 induces contraction, proliferation, and collagen synthesis of hepatic stellate cells in vitro, which may be mediated via the endothelin A receptor. It is unknown if specific blockade of the endothelin A receptor inhibits hepatic fibrosis in vivo, Methods: Groups of 10-20 rats with bile duct occlusion were treated with the nonpeptide endothelin-A receptor antagonist LU 135252 at 80 mg . kg(-1) . day(-1) from week 1-6 or from week 4-6, or with LU at 10 mg . kg(-1) . day(-1) from week 1-6, Animals with bile duct occlusion alone and sham-operated rats without or with LU at 80 mg . kg(-1) . day(-1) over 6 weeks sewed as controls. After 6 weeks, parameters of fibrogenesis were determined. Results: LU treatment led to improved histology, paralleled by a dose-dependence up to 60% reduction of liver collagen, even when administered at an advanced fibrosis stage, This was accompanied by a decreased messenger RNA of hepatic procollagen alpha 1(I) and tissue inhibitor of metalloproteinase 1, 2 major effecters of fibrosis, and of serum procollagen type III, a surrogate marker of liver fibrogenesis, Conclusions: Selective endothelin-A receptor blockade can dramatically reduce collagen accumulation in rat secondary biliary fibrosis, a model refractory to most potential antifibrotic agents. Endothelin-A receptor antagonists are promising antifibrotic agents in chronic liver disease.

关键词：投药口服丙氨酸转氨酶/血液碱性磷酸酶/血液腹水/药物疗法腹水/代谢天冬氨酸氨基转移酶类/血液胆红素/血液胆汁淤积/药物疗法胆汁淤积/代谢胆汁淤积/病理学胶原/分析胶原/生物合成 DNA 互补疾病模型动物内皮缩血管肽1/分析羟脯氨酸/分析高血压门静脉/药物疗法高血压门静脉/代谢高血压门静脉/病理学黄疸/药物疗法黄疸/代谢肝/药物作用肝/代谢肝/病理学肝硬化实验性/药物疗法肝硬化实验性/代谢肝硬化实验性/病理学器官测量丙酸苯酯类/药理学嘧啶类/药理学 RNA 信使/分析大鼠 Wistar 受体内皮素A 受体内皮素B 受体内皮缩血管肽/分析受体内皮缩血管肽/代谢金属蛋白酶1组织抑制剂/遗传学动物女(雌)性大鼠

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