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Clinical significance of plasma bone-specific alkaline phosphatase measurement and the alkaline phosphatase isozymes expression in osteosarcoma.

by Au Sze Ki. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves xii-xix). / Acknowledgement --- p.i / Table of Content --- p.ii / List of Abbreviation --- p.vi / Abstract --- p.viii / Chapter Chapter One : --- Introduction / Chapter 1.1. --- Osteosarcoma --- p.1 / Chapter 1.1.1. --- Definition --- p.1 / Chapter 1.1.2 --- "Incidence, geographic patterns of distribution and epidemiological consideration" --- p.1 / Chapter 1.1.3 --- "Age, sex and sites" --- p.3 / Chapter 1.1.4 --- Type and grade --- p.5 / Chapter 1.1.4.1. --- Grade --- p.5 / Chapter 1.1.4.2. --- Site --- p.5 / Chapter 1.1.4.3. --- Metastasis --- p.5 / Chapter 1.1.5 --- Histological features --- p.9 / Chapter 1.1.6 --- Clinical features --- p.9 / Chapter 1.1.7 --- Radiological features --- p.11 / Chapter 1.1.8. --- Molecuar genetics --- p.13 / Chapter 1.1.9 --- Treatment --- p.13 / Chapter 1.2 --- Biochemical Markers of Osteosarcoma --- p.14 / Chapter 1.2.1 --- Tumor marker --- p.14 / Chapter 1.2.2 --- Biochemical markers of bone turnover --- p.15 / Chapter 1.2.3 --- Change of biochemical marker in osteosarcoma --- p.17 / Chapter 1.3 --- Alkaline Phosphatase (ALP) --- p.17 / Chapter 1.3.1 --- ALPs Family --- p.20 / Chapter 1.3.2 --- Membrane binding --- p.22 / Chapter 1.3.3 --- Biochemical function and physiological role of ALP --- p.24 / Chapter 1.4 --- Normal values of serum ALP --- p.28 / Chapter 1.5 --- Clinical applications of ALP --- p.28 / Chapter 1.6 --- "Separation, identification and quantification of ALP isozymes" --- p.31 / Chapter 1.6.1 --- Themostability --- p.31 / Chapter 1.6.2 --- Inhibition studies --- p.31 / Chapter 1.6.3 --- Electrophoresis --- p.33 / Chapter 1.6.4 --- Isoelectric focusing --- p.34 / Chapter 1.6.5 --- Affinity precipitation --- p.34 / Chapter 1.6.6 --- Immunological studies --- p.35 / Chapter 1.7 --- Plasma BALP level as biochemical marker of osteosarcoma --- p.35 / Chapter 1.8 --- ALP in malignancies --- p.37 / Aim of study --- p.x / Chapter Chapter Two : --- Methods and Materials / Chapter 2.1 --- Plasma BALP measurement as a biochemical markerin osteosarcoma --- p.40 / Chapter 2.1.1 --- Patient groups --- p.40 / Chapter a) --- Normal subjects --- p.40 / Chapter b) --- Osteosarcoma patients --- p.40 / Chapter 2.1.2 --- Collection and preparation of patient bloods samples of patients --- p.40 / Chapter 2.1.3 --- Plasma total ALP measurement --- p.41 / Chapter a) --- Reagent --- p.41 / Chapter b) --- Procedure --- p.43 / Chapter 2.1.4 --- Plasma BALP measurements --- p.43 / Chapter a) --- Wheat germ lectin precipitation of BALP --- p.44 / Chapter i) --- Reagent / Chapter ii) --- Procedure / Chapter b) --- ABBOTT methods for plasma BALP activity measurement --- p.45 / Chapter c) --- COBAS MIRA methods for BALP measurement --- p.45 / Chapter d) --- ALKPHASE-B method of BALP measurement --- p.46 / Chapter 2.1.5 --- Inter-conversion of plasma BALP activity measurement in different methods --- p.47 / Chapter 2.1.6 --- Statistical analysis --- p.48 / Chapter 2.2 --- Alkaline phosphatase isozymes expression in human osteosarcoma --- p.48 / Chapter 2.2.1 --- In Vitro cultures of human SaOS-2 and U-2 OS osteosarcoma cell line --- p.48 / Chapter a) --- Reagent --- p.49 / Chapter b) --- Procedure --- p.50 / Chapter i) --- Storage of U-2 OS and SaOS-2 / Chapter ii) --- Subculture of confluent monolayer / Chapter 2.2.2 --- Protein assay --- p.51 / Chapter a) --- Standard Assay --- p.51 / Chapter i) --- Reagent / Chapter ii) --- Procedure / Chapter b) --- Mircoassay --- p.51 / Chapter i) --- Reagent / Chapter ii) --- Procedure / Chapter 2.2.3 --- Extraction of ALP from the cultured osteosarcoma cells --- p.52 / Chapter a) --- Reagent --- p.52 / Chapter b) --- Procedure --- p.52 / Chapter 2.2.4 --- "ALP extraction from human liver, placenta and osteosarcoma tissue" --- p.53 / Chapter a) --- Reagent --- p.53 / Chapter b) --- Procedure --- p.54 / Chapter 2.2.5 --- Isoelectric focusing of ALP --- p.55 / Chapter a) --- Preparation of the agarose IEF gel --- p.55 / Chapter b) --- Samples preparation --- p.56 / Chapter c) --- Isoelectric focusing --- p.57 / Chapter d) --- Protein detection --- p.59 / Chapter i) --- Reagent / Chapter ii) --- Procedure / Chapter e) --- Visualization of ALP isozyme --- p.60 / Chapter 2.2.6 --- Biochemical differentiation of ALP expressed in human osteosarcoma --- p.61 / Chapter a) --- Thermodenaturation of ALP --- p.61 / Chapter b) --- Ammino acid inhibition of ALP --- p.61 / Chapter 2.2.7 --- Immunohistostaining of placental ALP in human Osteosarcoma --- p.62 / Chapter a) --- Reagent --- p.62 / Chapter b) --- Preparation of human osteosarcoma cell line --- p.63 / Chapter c) --- Preparation of human osteosarcoma tissue --- p.63 / Chapter d) --- Immunohistostaining --- p.64 / Chapter Chapter Three : --- Results / Chapter 3.1 --- General information of the patients --- p.65 / Chapter 3.1.1 --- Age and sex distribution --- p.65 / Chapter 3.1.2 --- Sites --- p.65 / Chapter 3.1.3 --- Treatment and survival rate --- p.66 / Chapter 3.2 --- Clinical significance of plasma bone-specific alkaline phosphatase (BALP) activity measurementin osteosarcoma patients --- p.71 / Chapter 3.2.1 --- Plasma BALP activity measurement --- p.71 / Chapter 3.2.2 --- Normal reference of plasma BALP determination --- p.72 / Chapter 3.2.3 --- Diagnostic value of plasma BALP measurement in osteosarcoma --- p.75 / Chapter a) --- Plasma BALP level at admission --- p.75 / Chapter b) --- Plasma Total ALP level at admission --- p.78 / Chapter 3.2.4 --- Prognosis value of plasma BALP measurement in osteosarcoma Patients --- p.78 / Chapter a) --- Correlation of plasma BALP-Adm with the local relapse of the disease --- p.78 / Chapter b) --- Correlation of plasma BALP-Adm with survival rate of the patients --- p.90 / Chapter i) --- One year survival Rate / Chapter ii) --- two-year survival Rate / Chapter iii) --- Three-year survival rate / Chapter c) --- Correlation of the plasma BALP-Adm with the tumor volume --- p.90 / Chapter 3.2.5 --- Using plasma BALP measurement for monitoring of the disease --- p.91 / Chapter a) --- Effectiveness of pre-operative chemotherapy --- p.91 / Chapter b) --- Change of plasma BALP level during the treatment --- p.92 / Chapter i) --- Monitoring of pre-operative chemotherapy / Chapter ii) --- Detection of local recurrence and secondary metastasis / Chapter 3.3 --- Alkaline phosphatase isozyme expressionin osteosarcoma --- p.103 / Chapter 3.3.1 --- Isoelectric point (pI) gradient in isoelectric focusing (IEF)gel --- p.10? / Chapter 3.3.2 --- ALP isozyme standard --- p.103 / Chapter 3.3.3 --- Ectopic expression of ALP in human osteosarcoma cell line: U-2 OS and SaOS-2 --- p.104 / Chapter a) --- Isoelectric focusing separation --- p.104 / Chapter b) --- Biochemical differentiation of ALP extracts --- p.110 / Chapter 3.3.4 --- Alkaline phosphatase expression in osteosarcoma patient plasma sample --- p.110 / Chapter 3.3.5 --- Alkaline phosphatase isozyme expression in human osteosarcoma biopsy tissue --- p.111 / Chapter 3.3.6 --- Ectopic expression of placental ALP in human osteosarcoma by immunohistochemistry --- p.111 / Chapter a) --- Ectopic expression of placental ALP in human osteosarcoma cell line U-2 OS --- p.111 / Chapter b) --- Ectopic expression of placental ALP in human osteosarcoma tissue sections --- p.112 / Chapter Chapter Four : --- Discussion --- p.128 / Chapter Chapter Five : --- Conclusion --- p.142 / Bibliography --- p.xii / Appendix --- p.xx

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_322714
Date January 1997
ContributorsAu, Sze Ki., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish
Detected LanguageEnglish
TypeText, bibliography
Formatprint, ix, 144, (xii-xxi) leaves : ill (some col. mounted) ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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