by Paul, Liu Po Lung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 86-94). / ACKNOWLEDGMENT --- p.i / TABLE OF CONTENT --- p.ii / "LIST OF TABLE, FIGURE & PHOTO" --- p.viii / ABSTRACT --- p.x / Chapter CHAPTER ONE : --- INTRODUCTION --- p.1 / Chapter 1.1 --- ALKALINE PHOSPHATASE / Chapter 1.1.1 --- Alkaline Phosphatase Isoenzyme --- p.2 / Chapter 1.1.2 --- The Properties of Alkaline Phosphatases --- p.4 / Chapter 1.1.3 --- Serum Alkaline Phosphatases --- p.6 / Chapter 1.1.3.1 --- Placental Alkaline Phosphatase --- p.7 / Chapter 1.1.3.2 --- Intestinal Alkaline Phosphatase --- p.7 / Chapter 1.1.3.4 --- Skeletal Alkalne Phosphatase --- p.8 / Chapter 1.1.3.5 --- Hepatic Alkaline Phosphatase --- p.8 / Chapter 1.1.3.3 --- Renal Alkaline Phosphatase --- p.9 / Chapter 1.1.3.6 --- Miscellaneous Alkaline Phosphatase --- p.9 / Chapter 1.1.4 --- Problems in Discriminating the Skeletal and Hepatic Alkaline Phosphatase in Serum --- p.11 / Chapter 1.1.5 --- Quantitative measure of the Bone-Specific Alkaline Phosphatase --- p.12 / Chapter 1.1.6 --- Qualitative Detection of ALP isoenzymes --- p.14 / Chapter 1.2 --- OSTEOSARCOMA --- p.17 / Chapter 1.2.1 --- Definition --- p.17 / Chapter 1.2.2 --- Epidemiology and Statistics --- p.17 / Chapter 1.2.3 --- Clinical Presentation --- p.18 / Chapter 1.2.4 --- Radiographic finding --- p.19 / Chapter 1.2.5 --- Staging of Musculoskeletal Neoplasms --- p.20 / Chapter 1.2.6 --- Treatment of osteosarcoma --- p.21 / Chapter 1.2.6.1. --- Chemotherapy in Prince of Wales Hospital --- p.21 / Chapter 1.3 --- PLASMA AND TISSUE ALKALINE PHOSPHATASE IN NORMAL AND NEOPLASTIC CONDITION --- p.23 / Chapter 1.3.1 --- Normal values of plasma alkaline phosphatase --- p.23 / Chapter 1.3.2 --- Clinical use of elevated plasma & tissue alkaline phosphatase level in neoplastic conditions --- p.25 / Chapter 1.3.2.1 --- Helping the Diagnosis of the Osteosarcoma --- p.25 / Chapter 1.3.2.2 --- Monitoring the effect of chemotherapy --- p.26 / Chapter 1.3.2.3 --- Predicting the clinical course --- p.26 / Chapter 1.3.3 --- Qualitative measurement of ALP in plasma and tissue extract of osteosarcoma patient --- p.29 / Chapter 1.4 --- AIM AND SCOPE OF THE PRESENT DISSERATION --- p.30 / Chapter CHAPTER TWO : --- MATERIALS AND METHODS --- p.32 / Chapter 2.1 --- DIFERENT GROUPS OF PATIENTS --- p.33 / Chapter 2.1.1 --- Monitering the plasma bone specific ALP --- p.33 / Chapter 2.1.1.1 --- Osteosarcoma group --- p.33 / Chapter 2.1.1.2 --- Benign bone tumour group --- p.34 / Chapter 2.1.1.3 --- Metastasis group --- p.34 / Chapter 2.1.2 --- Collection of plasma samples preserve of tumor tissue --- p.34 / Chapter 2.2 --- QUANTITATIVE ANALYSIS OF THE PLASMA AND TISSUE BONE SPECIFIC ALKALINE PHOSPHATASE --- p.36 / Chapter 2.2.1 --- Extraction of tissue ALP --- p.36 / Chapter 2.2.1.1. --- Reagent --- p.36 / Chapter 2.2.1.2. --- Homogenization of the bone tissue --- p.36 / Chapter 2.2.1.3. --- Extraction of ALP --- p.37 / Chapter 2.2.2 --- Assay for Bone-specific ALP --- p.38 / Chapter 2.2.2.1. --- Reagents --- p.38 / Chapter 2.2.2.2. --- Procedures --- p.38 / Chapter 2.3 --- QUALITATIVE MEASUREMENT OF ALP ISOENZYME --- p.40 / Chapter 2.3.1 --- Equipment required --- p.40 / Chapter 2.2.2 --- Practical procedure --- p.40 / Chapter 2.3.3.1 --- Gel casting --- p.40 / Chapter 2.3.3.2 --- Sample preparation and application --- p.42 / Chapter 2.3.3.3 --- Electrofocusing --- p.42 / Chapter 2.3.3.4 --- Western blotting of the protein --- p.43 / Chapter 2.3.3.5 --- Detection methods --- p.45 / Chapter 2.4 --- METHOD OF STATISTICAL ANALYSIS --- p.48 / Chapter CHAPTER THREE : --- RESULTS --- p.49 / Chapter 3.1 --- QUANTITATIVE MEASUREMENT OF PLASMA AND TISSUE BONE SPECIFIC ALKALINE PHOSPHATASE --- p.50 / Chapter 3.1.1 --- General Information of the patients monitoring --- p.50 / Chapter 3.1.2 --- Pretreatment evaluation --- p.52 / Chapter 3.1.3 --- Correlation between the pretreatment plasma ALP levels and prognosis in the osteosarcoma patient group --- p.57 / Chapter 3.1.4 --- "Correlation between the pre-operational, post- operational plasma ALP levels and the prognosis of osteosarcoma" --- p.59 / Chapter 3.1.5 --- Analysis of plasma ALP levels at the time of relapse in osteosarcoma patients --- p.61 / Chapter 3.1.6 --- Usefulness of the plasma ALP levels for monitoring the effectiveness of chemotherapy --- p.62 / Chapter 3.1.7 --- Correlation between the ALP levels in the tumor extract and the prognosis of the osteosarcoma --- p.64 / Chapter 3.2 --- QUALITATIVE ANALYSIS OF THE PLASMA AND TISSUE ALKALINE PHOSPHATASE LEVEL --- p.67 / Chapter 3.2.1 --- Comparison of the result of Isoelectric focusing of the plasma ALP of the osteosarcoma patients and the normal subjects --- p.67 / Chapter 3.2.1 --- Result of Isoelectric focusing of the ALP isoenzymes in the tissue extract of the osteosarcoma and normal bone --- p.70 / Chapter CHAPTER FOUR : --- DISCUSSION --- p.72 / Chapter 4.1 --- USE OF QUANTITATIVE MONITORING OF PLASMA ALP AND MEASURING TISSUE ALP IN OSTEOSARCOMA PATIENTS --- p.73 / Chapter 4.2 --- ISOELECTRIC FORCUSING AS A TECNIQUE FOR QUALITATIVE MEASUREMENT OF PLASMA AND TISSUE ALKALINE PHOSPHATASE --- p.80 / Chapter CHAPTER FIVE : --- CONCLUSION --- p.83 / Chapter CHAPTER SIX : --- BIBILOGRAPHY --- p.85

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

D.Litt. et Phil. ( Psychology) / A literature survey has confirmed that the growing number of cancer survivors in general are experiencing both physical as well as psychological and social adaptation difficulties following their cancer treatment. This is no different for patients diagnosed with Osteosarcoma and Ewing sarcoma (bone sarcomas) during adolescence. At present there is no formal psychosocial intervention program available to the patients presenting with Ewing and Osteosarcomas. Because of the multicultural nature of the patients as well as the fact that many childhood and adolescent cancers do not include major surgery, as is the case with bone sarcomas, the direct applicability of generalized psychosocial intervention programs for adolescents with cancer other than bone sarcoma is doubtful ...