Objetivo: Foi investigada a hipótese da hiperprolactinemia modular a resposta inflamatória alérgica pulmonar em ratos machos e em fêmas lactantes sem tratamento de domperidona. Métodos: Em ratos machos, a hiperprolactinemia foi de curta duração (5 dias) induzida pela domperidona (5,1 mg.kg-1 por dia, i.p). A resposta alérgica foi gerada por sensibilização e desafios inalatórios com ovoalbumina. Foi feita contagem de leucócitos totais e diferenciados do lavado bronco alveolar (BAL), lavado medular femoral (BFL) e sangue; a percentagem de produção de muco e colageno no pulmão, níveis de corticosterona e prolactina e citocinas TNF-α, IL-4, IL-6, IL-10, em explantes de pulmão e IFNg no BAL, foram medidos. Pela citometria foram avaliadaos os receptores de prolactina; Resultados: Hiperprolactinemia de curta duração feita antes do desafio inalatório disminuiu a resposta alérgica pulmonar na contagem de leucócitos no lavado broncoalveolar. Esse tratamento reduziu a celularidade no BFL e a percentagem de muco e aumentou a expressão de citocinas IL-4, IL-6, IL-10, TNFα e da expressão do IFNg. Níveis altos de prolactina diminuiram o número de eosinófilos ao pulmão no BAL. Pela citometria revelou-se que além de ter menor número de granulócitos migrados ao pulmão, estes apresentaram maior expressão do número de receptores por granulócito para prolactina no grupo tratado com domperidona. Alterações similares foram reveladas em fêmeas lactantes como foi a diminuição nos leucócitos do BAL, e no número de células do BFL. O tratamento profilático diminuiu a resposta alérgica tanto no grupo hiperprolactinêmico como no grupo veículo. O tratamento feito após o desafio inalatório não evidenciou alterações relevantes nas variáveis medidas. Conclusões: A hiperprolactinemia de curta duração, feita após a sensibilização e antes da inalação diminui a resposta inflamatória no pulmão em ratos. Os resultados deste estudo demonstram que a hiperprolactinemia induzida antes do desafio antigênico diminue a inflamação alérgica pulmonar. Assim, é provável que a prolactina endógena tenha um papel relevante como um imunomodulador da asma. Este estudo aponta a possibilidade futura do uso da domperidona para pacientes asmáticos. Durante a primavera muitas espécies de mamíferos têm seus filhotes e ocorre abundância de fatores alergenos no ar. Logo, um fator endógeno que favoreça a proteção de fêmeas durante a lactação, tal como a hiperprolactinemia, tem elevado valor adaptativo / Objective: It was investigated if hyperprolactinemia has modulatory actions on lung allergic inflammatory response in male rats. Lactating female rats were tested for pulmonary allergy as well. Methods: In male rats, short-term (5 days) hyperprolactinemia was induced by domperidone (5.1 mg.kg-1 per day, ip). Allergic response was generated by sensitization and inhalation challenge with ovalbumin. Total and differential leukocytes bronchoalveolar lavage (BAL), femoral medullary lavage (BFL) and blood; the percentage of collagen and mucus production in the lungs, plasma levels of corticosterone and prolactin cytokines and TNF-α, IL-4, IL-6, IL-10, IFNg explants lung and BAL, were measured. Flow cytometry was used to evaluate prolactin receptor; Results: Short-term hyperprolactinemia made before the inhaled challenge reduced the pulmonary allergic response in white blood cell counts in BAL. This treatment reduced the cellularity in BFL and the percentage of mucus and increased expression of cytokines IL-4, IL-6, IL-10, TNFa and IFNg expression. High prolactin levels decreased the number of eosinophils to the lung in BAL. There were fewer granulocytes migrated to the lung. These granulocytes showed higher expression prolactin receptors in hyperprolactinemia animals. Similar changes were revealed in lactating females. In these animals, there was a reduction in BAL leukocyte, and the number of cells BFL. Prophylactic treatment decreased the allergic response in both hyperprolactinemic and vehicle groups. The treatment made after inhalational challenge did not induce significant changes in the variables measured in this study. Conclusions: Short-term hyperprolactinemia, made after sensitization and before inhalation, decreases the inflammatory response in the lung of rats. The results of this study demonstrate that hyperprolactinemia, induced before antigen challenge, decreases pulmonary allergic inflammation. Thus, it is probable that the endogenous prolactin has an important role as an immunomodulator of asthma. This study points out the prospect of a future use of domperidone for asthmatic patients. For various mammalian species, parturition occurs during springtime. Pollen in the air might be an abundant allergic factor during springtime. Thus, protecting lactating females against this type allergy might have high adaptive value

Gu Jia-Shi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 117-148). / Abstracts in English and Chinese. / Title --- p.i / Abstract (English) --- p.iii / Abstract (Chinese) --- p.iv / Acknowledgments --- p.v / Statement of Originality --- p.vi / List of Abbreviations --- p.viii / List of Figures --- p.xi / List of Tables --- p.xiii / Table of Contents --- p.xiv / Chapter Section One : --- Introduction & Literature Review / Introduction & Objective --- p.2 / Introduction / Objective of the present study / Chapter Chapter. 1 --- A Review of Chronic lung disease (CLD) --- p.6 / Chapter 1. --- BPD 226}0ؤ an example of CLD / Chapter 2. --- Pathological change & Clinical presentation / Chapter 3. --- Clinical sequel of CLD infants / Chapter 3.1 --- O2 consumption of CLD infants / Chapter 3.1-1 --- Oxygen consumption / Chapter 3.1-2 --- Oxygen transportation / Chapter 3.1-2a --- Dissolved O2 / Chapter 3.1-2b --- Haemoglobin / Chapter 3.2 --- Energy expenditure of CLD infants / Chapter 3.3 --- Growth rate of CLD infants / Chapter 4. --- Treatment & Management of CLD infants / Chapter 4.1 --- Diuretics / Chapter 4.2 --- Bronchodilators / Chapter 4.3 --- Corticosteroids / Chapter 5. --- "Interpretations of the observed phenomena, why does CLD impair growth?" / Chapter 5.1 --- The traditional view / Chapter 5.2 --- Disagreement with the traditional view / Chapter Chapter 2 --- Measurement of oxygen consumption --- p.20 / Chapter 1. --- Invasive measurement of VO2 / Chapter 1.1 --- Cardiac output / Chapter 1.2 --- Fick method / Chapter 1.3 --- Advantages and Disadvantages of Fick method in estimating VO2 / Chapter 1.4 --- Measurement of cardiac output by thermodilution / Chapter 1.4-1 --- Advantages and Disadvantages of Thermodilution Method / Chapter 2. --- Non-invasive measurement of VO2 / Chapter 2.1 --- Metabolic analyzer---DeltatraćёØII / Chapter 2.2 --- Paramagnetic sensor / Chapter 3. --- Measured and calculated oxygen consumption / Chapter 3.1 --- Difference between mVO2 and cVO2 / Chapter 4. --- Summary / Chapter Chapter 3 --- Hypothesis --- p.34 / Chapter 1. --- Possible explanations for the difference between mV02 & cV02 / Chapter 1.1 --- Measurement variability and Mathematical error / Chapter 1.2 --- Oxygen consumption of the lung / Chapter 1.3 --- VO2pul with lung damage / Chapter 1.4 --- "Neutrophils, Macrophages and oxygen consumption" / Chapter 2. --- Hypothesis / Chapter Section Two : --- Methods & Materials / Chapter Chapter 1 --- Animal Model --- p.41 / Chapter Chapter 2. --- Materials --- p.43 / Chapter 1. --- Animals / Chapter 2. --- Chemicals used for inducing lung damage / Chapter 2.1 --- Acute damage group / Chapter 2.1-1 --- N-nitroso-N-methylurethane (NNNMU) / Chapter 2.1-2 --- Administrations to rabbits / Chapter 2.2 --- Chronic damage group / Chapter 2.2-1 --- Bleomycin (BLM) / Chapter 2.2-2 --- Pulmonary toxicity of Bleomycin / Chapter 2.2-3 --- Administration to animals / Chapter Chapter 3 --- Instruments --- p.50 / Chapter 1. --- Measurement of VO2 and VCO2 226}0ؤDeltatracIÍёØ Metabolic analzyer / Measurement of cardiac outpu´tؤCardiomax II model85 / Chapter Chapter 4 --- Methods --- p.58 / Chapter 1. --- N-nitroso-N-methylurethane (NNNMU) Preparation / Chapter 2. --- Bleomycin Preparation / Chapter 3. --- 2.5% pentobarbitone Preparation / Chapter 4. --- Animal Preparation / Chapter 4.1 --- Control (Normal) group / Chapter 4.2 --- A cute lung damage group / Chapter 4.3 --- Chronic lung damage group / Chapter 5. --- Preparation of the animals for VO2 measurement / Chapter 6. --- Measurement of oxygen consumption / Chapter 6.1 --- VO2wb measurement / Chapter 6.2 --- VO2b measurement / Chapter 7. --- Histopathology / Chapter 8. --- Statistics / Chapter Section Three : --- Results --- p.69 / Chapter 1. --- Healthy (Control) group / Chapter 1.1 --- Pulmonary histology / Chapter 2. --- Acute lung damage group / Chapter 2.1 --- Pulmonary histology / Chapter 3. --- Chronic lung damage group / Chapter 3.1 --- Pulmonary histology / Chapter 4. --- Comparison of the pulmonary oxygen consumption among the three groups / Chapter Section Four : --- Discussion --- p.97 / Chapter Section Five : --- Conclusion --- p.111 / Chapter Section Six : --- Future Studies --- p.114 / Chapter Section Seven : --- Bibliography --- p.118

The bronchiectatic lung is a diseased state in which the airways are chronically damaged and dilated. This state is found in the clinical entities of cystic fibrosis and non-cystic fibrosis bronchiectasis. These are two highly relevant chronic suppurative lung diseases in which an understanding of the microbiology of these patients is considered key to appropriate management. This has traditionally been via the use of traditional culture techniques. However, with the development of molecular methodologies, the previously perceived wisdom is being challenged. In both cystic fibrosis and non-cystic fibrosis bronchiectasis, Pseudomonas aeruginosa is considered the most significant pathogen. In CF there has been considerable concern surrounding the risk of transmission of Pseudomonas aeruginosa between patients on the basis of a significant quantity of research into this matter. In contrast, there has been very little research performed into the equivalent risk in non-cystic fibrosis bronchiectasis. In this thesis we describe an extensive single-centre epidemiological review of Pseudomonas aeruginosa spanning both these diseases. Via this we have shown evidence of cross-infection within a non-cystic fibrosis bronchiectasis cohort. This epidemiological review has included multiple genotyping methods including multilocus sequence typing and whole genome sequencing, As an extension of the epidemiological review, we have performed an in silico prediction of hypermutator status from the whole genome sequencing data to provide greater understanding of the likelihood of cross-infection, and have also demonstrated a culture-independent adaption of multilocus sequence typing for potential screening for cross-infection. In addition to Pseudomonas aeruginosa, we have also looked at the wider bacterial community in the lungs of patients with these two conditions via culture-independent techniques. We have shown that whilst Pseudomonas aeruginosa is often an important component, these are clearly complex communities. We have primarily investigated the cohort with non-cystic fibrosis bronchiectasis, but we have demonstrated associations between clinically-relevant markers and complexity of the bacterial communities within the lungs of both these cohorts of patients. Whilst we have used the gold-standard technique of 16S rRNA sequencing, we have also shown the validity of a simple and potentially more feasible profiling technique for standard clinical care. In summary, through the application of culture-dependent and independent molecular techniques, this research has shed light on the epidemiology of Pseudomonas aeruginosa within our respiratory cohorts, and the complexity and clinical relevance of the wider microbial communities within these patients. Such studies are essential if we are to advance our understanding of the bronchiectatic lung and optimise strategies for patient management.

Background: Lung cancer was the fifth leading cause of mortality globally in 2010, and the leading cause of cancer mortality in Canada, representing 26% of all cancer deaths for both men and women in 2017. Radon is a very modifiable environmental exposure that is the second most important cause of lung cancer. Objectives: The objectives of this thesis are to quantify the lung cancer burden associated with residential radon and to identify the most cost effective mitigation options to reduce residential radon in Canada. Methods: The global burden of lung cancer mortality attributable to radon in 2012 was estimated from the 66 countries for which a representative national radon survey was available, using several different models for excess relative risk (ERR) of lung cancer from radon studies. Cost-utility analyses are conducted for 20 practical radon interventions scenarios to reduce residential radon exposures in new and existing housing in Canada, each province/territory and 17 census metropolitan areas. A societal perspective and a lifetime horizon are adopted. A Markov cohort model and a discrete event simulation are used to model residents by household, based on a period-life table analysis, at a discount rate of 1.5%. Results: The estimates of the global median PAR were consistent, ranging from 16.5% to 13.6% for the three ERR models based on miners, and the mean estimates of PAR for Canada ranged from 16.3% to 14.6%. It is very cost effective to install radon preventive measures in new construction compared to no radon control in all regions across Canada. At a radon mitigation threshold of 100 Bq/m3, the sequential analysis recommends the combination of the activation of preventive measures in new housing with the mitigation of existing housing at current testing and mitigation rates for cost effectiveness thresholds between 51,889 and 92,072 $/QALY for Canada, between 27,558 and 85,965 $/QALY for Manitoba, and between 15,801 and 36,547 $/QALY for the Yukon. The discounted ICER for screening and mitigation of existing housing at current rates relative to no radon control measures is 62,451 (66,421) $/QALY using a Markov cohort model (discrete event simulation model) for mitigation of housing above a threshold of 200 Bq/m3, and is 58,866 (59,556) $/QALY using a Markov cohort model (discrete event simulation model) for mitigation of housing above a threshold of 100 Bq/m3. Conclusions: Cost effective residential radon interventions should be implemented across Canada to reduce exposures to this very modifiable cause of lung cancer and to help reduce the increasing lung cancer burden in an ageing Canadian population.

Objetivos: Desenvolver e caracterizar nanocápsulas poliméricas contendo erlotinib, bem como avaliar sua atividade antitumoral in vitro em células de adenocarcinoma de pulmão humano. Metodologia: As nanocápsulas contendo erlotinib (0,5 mg.mL-1) foram obtidas pelo método de nanoprecipitação utilizando poli(ε-caprolactona) e óleo de copaíba como parede polimérica e núcleo oleoso, respectivamente. Os parâmetros físico-químicos avaliados foram: diâmetro médio e distribuição de tamanho, índice de polidispersão, potencial zeta, concentração de partículas e pH. Para determinação do teor e eficiência de encapsulação do erlotinib, utilizou-se metodologia validada por CLAE-UV. O estudo de liberação in vitro, utilizando sacos de diálise, foi realizado para obter o perfil de liberação do fármaco a partir das nanocápsulas. As nanocápsulas contendo erlotinib foram avaliadas quanto ao seu potencial de inibir o crescimento, induzir a apoptose, interferir com o ciclo celular e sobrevivência clonogênica de células de adenocarcinoma de pulmão, linhagem A549. Resultados: As nanocápsulas apresentaram diâmetro médio de 171 ± 2 (PDI < 0, 10), potencial zeta de −8,17 ± 2.26 mV, número de partículas por mL de 6,97 ± 0,22 × 1013 e pH de 6,24 ± 0,02. O teor e a eficiência de encapsulação foram próximos de 100%. Com exceção do pH, todos parâmetros mantiveram-se iguais após 30 dias de armazenamento em temperatura ambiente. Observou-se uma liberação controlada do fármaco devido à nanoencapsulação. Os ensaios de citotoxicidade demonstraram que as nanocápsulas contendo erlotinib apresentaram maior atividade antitumoral quando comparado com o fármaco livre. Também foi demonstrado indução de apoptose, pela análise de ciclo celular e marcação por Anexina-V conjugada ao 7-AAD. No ensaio clonogênico, as nanocápsulas contendo erlotinib reduziram 100% o número de colônias formadas. Conclusões: Foram obtidas nanocápsulas com propriedades nanotécnologicas adequadas e capazes de controlar a liberação do erlotinib. Os estudos in vitro na linhagem celular A549 demonstraram aumento no efeito antitumoral e foi demonstrado que o encapsulamento do fármaco é imprescindível para essa melhor atividade. / Purpose: To develop and characterise erlotinib-loaded polymeric nanocapsules and to evaluate its in vitro antitumor activity in human lung adenocarcinoma cells. Methodology: The erlotinib-loaded nanocapsules (0.5 mg.mL-1) were obtained by nanoprecipitation method using poly (ε-caprolactone) and copaiba oil as the polymeric wall and oily core, respectively. The physicochemical parameters evaluated were: mean diameter and size distribution, polydispersity index, zeta potential, particle concentration and pH. An HPLC-UV validated method was used to determine the drug content and encapsulation efficiency. The in vitro release study using dialysis bags was performed to obtain the drug release profile from nanocapsules. The erlotinib-loaded nanocapsules were evaluated regarding their potential to inhibit the growth, induce apoptosis, interfere with the cell cycle and clonogenic survival of lung adenocarcinoma cell (A549). Results: The nanocapsule formulation presented z-average diameter of 171 ± 2 (PDI <0.10), zeta potential value of -8.17 ± 2.26 mV, number of particles per mL of 6.97 ± 0.22 × 1013, and pH value of 6.24 ± 0.02. The drug content and the encapsulation efficiency were nearly 100%. Except for the pH value, all these parameters remained the same after 30 days of storage. A controlled release of the drug was observed due to nanoencapsulation. The cytotoxicity assays demonstrated that the erlotinib-loaded nanocapsules showed higher antitumor activity compared to free drug. Induction of apoptosis was demonstrated by cell cycle analysis and Annexin-V/7AAD staining. In the clonogenic assay, erlotinib-loaded nanocapsules reduced 100% the number of colonies formed. Conclusions: Nanocapsules with appropriate nanotechnological properties and capable of controlling the erlotinib release were obtained. The in vitro studies in the A549 cell line showed an increase in antitumor effect and was demonstrated that the drug encapsulation is essential for this better activity.

Xu Feng. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (p. 205-233). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Glioblastoma Multiforme (GBM) is an extremely aggressive grade IV brain tumour that is highly infiltrative and can spread to other parts of the brain quickly. It is the most common primary brain tumour where patients have a median survival of 14.6 months. Symptoms vary depending upon the location of the tumour and include seizures, progressive headaches and focal neurological deficit. The poor prognosis is characterised by deregulation of many key signalling pathways involving survival, growth, apoptosis and evasion of immune surveillance. In this study, we investigated whether complement factor H related protein 5 (CFHR5) from primary GBM cells direct from patients exhibited functional activity similar to factor H. The presence of CFHR5 was validated by western blot and ELISA technique from B30, B31 and B33 primary GBM cells. The functional capacity of CFHR5 was examined through the alternative pathway, co-factor, and decay acceleration assay. We demonstrated that CFHR5 was able to inhibit the alternative pathway through the same mechanism as factor H. Emerging evidence had shown that the innate immune protein surfactant protein D (SP-D) and recombinant human SP-D (rhSP-D) were able to induce apoptosis in eosinophilic leukaemic cells. We studied the ability of rhSP-D to induce apoptosis in U87 GBM cells through apoptotic and viability assays. rhSP-D was unable to mediate cell death and instead increased cell viability. This led us to investigate the expression of SP-D in U87 and B30 GBM cells through western blot, ELISA and immuno-fluorescence detection. We demonstrated novel information about the production of SP-D by GBM cells. To extend our study, we investigated the interaction of THP-1 macrophage with rhSP-D bound U87 cells. We carried out live cell imaging, RT-qPCR, and cell viability assays, to study the changes in cytokine expression and viability of cells. THP-1 did not engulf U87 cells; however, it did reduce the number of cells and decrease the expression of pro-tumourigenic cytokines. This study highlights the ability of primary GBM cells to evade innate immune detection by the secretion of functionally active CFHR5. It also demonstrated the ability of U87 to evade destruction by rhSP-D and THP-1 highlighting the extremely aggressive behaviour of the tumour and lack of new treatment to improve prognosis in over a decade.

Mechanical ventilation is vital for the treatment of patients in respiratory intensive care and can be life saving. However, the risks of regional pressure gradients and over-distension must be balanced with the need to maintain function. For these reasons mechanical ventilation can benefit from the regional information provided by bedside imaging such as electrical impedance tomography (EIT). In this thesis we develop and test methods to retrieve clinically meaningful measures of lung function from EIT and examine the feasibility of closing the feedback loop to enable EIT-guided control of mechanical ventilation. Working towards this goal we develop a reconstruction algorithm capable of providing fast absolute values of conductivity from EIT measurements. We couple the resulting conductivity time series to a compartmental ordinary differential equation (ODE) model of lung function in order to recover regional parameters of elastance and airway resistance. We then demonstrate how these parameters may be used to generate optimised pressure controls for mechanical ventilation that expose the lungs to minimal gradients of pressure and are stable with respect to EIT measurement errors. The EIT reconstruction algorithm we develop is capable of producing low dimensional absolute values of conductivity in real time after a limited additional setup time. We show that this algorithm retains the ability to give fast feedback on regional lung changes. We also describe methods of improving computational efficiency for general Gauss-Newton type EIT algorithms. In order to couple reconstructed conductivity time series to our ODE model we describe and test the recovery of regional ventilation distributions through a process of regularised differentiation. We prove that the parameters of our ODE model are recoverable from these ventilation distributions apart from the degenerate case where all compartments have the same parameters. We then test this recovery process under varying levels of simulated EIT measurement and modelling errors. Finally we examine the ODE lung model using control theory. We prove that the ODE model is controllable for a wide range of parameter values and link controllability to observable ventilation patterns in the lungs. We demonstrate the generation and optimisation of pressure controls with minimal time gradients and provide a bound on the resulting magnitudes of these pressures. We then test the control generation process using ODE parameter values recovered through EIT simulations at varying levels of measurement noise. Through this work we have demonstrated that EIT reconstructions can be of benefit to the control of mechanical ventilation.

肺癌是導致男性死亡的最常見原因以及是排在乳腺癌和結腸癌之後的導致女性死亡的第三大原因。雖然肺癌如此嚴重，但是如今治疗肺癌仍然是一个挑战。現今對肺癌的治療主要集中在化學治療和靶點藥物治療，但是由於這些治療有著很大的副作用和低治愈率，尋找其他的醫學替代方法十分迫切。甘草是其中最常用的中藥，它常常用作食品工業中的甜味劑。以往的研究表明，甘草具有多種的生物活性。但是甘草提取物對於肺癌的治療卻是十分匱乏的。 / 本論文主要目的是評價甘草提取物以及其中的有效成份對非小型肺癌細胞株A549 的影響，以及其作用的機理。我們的數據表明，甘草的乙酸乙酯（EAL）成份比甘草的乙醇提取物有著比較強的抑制癌細胞的作用。另外，對甘草的五個單體進行的測試中發現lico-3 是最具有抑制肺癌作用的。利用高效液相色譜法對甘草活性成份分析表明，lico-3 是EAL中的其中一個單體。 / 乳酸脫氫酶滲漏（LDH）的檢測結果以及异硫氰酸荧光素-碘化丙啶（FITC-PI）雙染的結果表明，EAL 能夠引起肺癌細胞的凋亡現象而非壞死現象。實驗結果表明由EAL引起的A549細胞凋亡是跟Bcl-2家族及Caspase家族有關係，同時EAL還能夠抑制Akt途徑從而導致細胞的死亡。 / 致肺癌細胞死亡的原因進行進一步研究表明，EAL還能夠引起抑制細胞週期的運作，停留在G2/M 時期。這可能是由於EAL引發了p53與p21的上調作用從而抑制了細胞的生長與增殖。 / 實驗結果說明了EAL引起的肺癌細胞株A549的凋亡作用是跟多重細胞通路有關， 同時表明了EAL是具有抗擊肺癌作用的潛能，能夠作為治療肺癌的藥物。 / Lung cancer is the most common cause of cancer death in men and third in women followed by breast cancer and colon cancer, yet treatment of lung cancer remains a challenge. Current treatments including chemotherapy and targeted drug treatment come with side-effects and low successful rate. Alternative medicine for treatment of lung cancer is warranted. Glycyrrhiza uralensis (Gan-Cao), commonly called “licorice, is one of the most commonly used herbs in traditional Chinese medicine (TCM). It is also used as flavoring and sweetening agents in many of food products. Previous studies have indicated that licorice exhibits a variety of biological activities. However, anticancer effects of licorice extract on lung cancer remain unclear. / In this study, we evaluated effects of licorice extract and its chemical components on human lung cancer cell line A549, and studied its mode of action. Our results showed the ethyl acetate fraction of licorice (EAL) was more effective in inhibition of A549 cell growth followed by ETL (IC₅₀: 50μg/mL). Moreover, among the five compounds tested, lico-3 was more potent compound. The HPLC analysis of the active fraction indicated that lico-3 was one of the compounds distributed in the EA fraction. / The results of LDH assay and FITC-PI co-staining method suggested low concentration of EAL can trigger apoptosis but not necrosis. The experimental findings show that EAL induce apoptosis in A549 cell lines involved in Bcl-2 family and caspase cascade. Also, EAL can arrest the Akt survival pathway in A549. Furthermore, the results indicate that EAL triggered G2/M phase arrest. The studies suggest EAL can up-regulate p53 and p21 to promote cell cycle arrest resulting in inhibition of proliferation. / Experimental results indicate that EAL is involved in multiple signal pathways to induce lung cancer cell death. The result suggests EAL is a potential candidate for lung cancer therapy. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Zhou, Yanling. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 99-110). / Abstracts in Chinese. / Abstract --- p.III / 論文摘要 --- p.V / Acknowledgement --- p.VII / List of Contents --- p.VIII / List of Figures --- p.X / List of Tables --- p.XI / List of Abbreviations --- p.XII / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Lung cancer --- p.1 / Chapter 1.1.1 --- Overview --- p.1 / Chapter 1.1.2 --- Risk factors --- p.2 / Chapter 1.1.3 --- Types of lung cancer --- p.4 / Chapter 1.1.4 --- Stages and treatment of lung cancer --- p.5 / Chapter 1.1.5 --- Chemotherapy for lung cancer treatment --- p.8 / Chapter 1.2 --- Traditional Chinese Medicines --- p.11 / Chapter 1.2.1 --- Overview --- p.11 / Chapter 1.2.2 --- Licorice --- p.14 / Chapter 1.2.3 --- Chemical study of licorice --- p.16 / Chapter 1.2.4 --- Pharmacological activities of licorice --- p.16 / Chapter 1.3 --- Molecular mechanism of apoptosis --- p.21 / Chapter 1.3.1 --- Overview --- p.21 / Chapter 1.3.2 --- Bcl2 family --- p.21 / Chapter 1.3.3 --- Caspase pathway --- p.23 / Chapter 1.3.4 --- Akt pathway --- p.24 / Chapter 1.3.5 --- p53 protein --- p.26 / Chapter 1.3.6 --- Apoptosis and cancer --- p.27 / Chapter 1.4 --- Cell cycle --- p.29 / Chapter 1.4.1 --- Overview --- p.29 / Chapter 1.4.2 --- Cell cycle and p53 --- p.29 / Chapter 1.4.3 --- Cell cycle and cancer --- p.30 / Chapter 1.5 --- Aims of study --- p.32 / Chapter Chapter 2 --- Materials and Methods --- p.33 / Chapter 2.1 --- Cell culture and treatment --- p.33 / Chapter 2.1.1 --- Cell line --- p.33 / Chapter 2.1.2 --- Chemicals and reagents --- p.34 / Chapter 2.1.3 --- Preparation of solutions --- p.34 / Chapter 2.2 --- Preparation of Licorice sample --- p.35 / Chapter 2.3 --- HPLC analysis --- p.35 / Chapter 2.3.1 --- Chemical and materials --- p.35 / Chapter 2.3.2 --- Instrumentation --- p.36 / Chapter 2.3.3 --- Preparation of Standard solutions --- p.36 / Chapter 2.3.4 --- Preparation of samples --- p.37 / Chapter 2.3.5 --- HPLC conditions --- p.37 / Chapter 2.3.6 --- Method validation --- p.37 / Chapter 2.4 --- Cell viable assay --- p.38 / Chapter 2.4.1 --- Samples preparation --- p.39 / Chapter 2.4.2 --- Procedure --- p.39 / Chapter 2.5 --- LDH assay --- p.40 / Chapter 2.5.1 --- Reagent preparation --- p.40 / Chapter 2.5.2 --- Procedure --- p.41 / Chapter 2.6 --- Annexin V assay --- p.41 / Chapter 2.6.1 --- Reagent --- p.42 / Chapter 2.6.2 --- Procedure --- p.42 / Chapter 2.7 --- Cell cycle study --- p.43 / Chapter 2.7.1 --- Chemicals and reagent --- p.43 / Chapter 2.7.2 --- Procedure --- p.44 / Chapter 2.8 --- Caspase3/7 Assay --- p.44 / Chapter 2.8.1 --- Reagent preparation --- p.45 / Chapter 2.8.2 --- Procedure --- p.46 / Chapter 2.9 --- Western blotting --- p.46 / Chapter 2.9.1 --- Reagent and antibodies --- p.46 / Chapter 2.9.2 --- Procedure --- p.50 / Chapter 2.9.3 --- Determination of protein concentration --- p.51 / Chapter 2.10 --- Data analysis --- p.51 / Chapter Chapter 3 --- Results --- p.52 / Chapter 3.1 --- Chromatographic conditions and HPLC identity conformation --- p.52 / Chapter 3.1.1 --- Linearity, limits of detection and quantification --- p.56 / Chapter 3.1.2 --- Reproducibility --- p.56 / Chapter 3.1.3 --- Analysis of ethyl acetate of licorice (EAL) using the validated method --- p.56 / Chapter 3.2 --- Licorice induces apoptosis in nonsmall cell lung carcinoma --- p.61 / Chapter 3.2.1 --- Cell viability assay --- p.61 / Chapter 3.2.2 --- LDH leakage assay --- p.71 / Chapter 3.2.3 --- Annexin V and PI staining --- p.73 / Chapter 3.3 --- Protein expression in EALinduced apoptotic cells --- p.75 / Chapter 3.3.1 --- Bcl2 family --- p.75 / Chapter 3.3.2 --- Activation of caspases by EAL treatment --- p.77 / Chapter 3.4 --- EAL could block Akt survival pathway --- p.79 / Chapter 3.5 --- EAL induces cell cycle arrest in nonsmall cell lung carcinoma --- p.83 / Chapter Chapter 4 --- Discussion --- p.85 / Chapter 4.1 --- Chemical analysis of licorice --- p.85 / Chapter 4.2 --- Licorice induced apoptosis but not necrosis on lung cancer cell A549 --- p.86 / Chapter 4.2.1 --- Licorice exhibits specific cytotoxicity to different cancer cells in vitro --- p.86 / Chapter 4.2.2 --- EAL induces cell death via apoptosis but not necrosis --- p.87 / Chapter 4.3 --- Growth inhibition by EAL inducing apoptosis --- p.89 / Chapter 4.3.1 --- EAL induces apoptotic cell death through modification of Bcl2 family --- p.89 / Chapter 4.3.2 --- EAL activate the caspase proteins --- p.90 / Chapter 4.4 --- Growth inhibition by EAL inducing survival pathway arrest --- p.92 / Chapter 4.5 --- Growth inhibition by EAL inducing cellcycle arrest --- p.94 / Chapter 4.6 --- General discussion --- p.96 / Reference --- p.99

With ever increasing advancements in imaging, there is an increasing abundance of images being acquired in the clinical environment. However, this increase in information can be a burden as well as a blessing as it may require significant amounts of time to interpret the information contained in these images. Computer assisted evaluation is one way in which better use could be made of these images. This thesis presents the combination of texture analysis of images acquired during the treatment of cancer with machine learning in order to improve radiotherapy. The first application is to the prediction of radiation induced pneumonitis. In 13- 37% of cases, lung cancer patients treated with radiotherapy develop radiation induced lung disease, such as radiation induced pneumonitis. Three dimensional texture analysis, combined with patient-specific clinical parameters, were used to compute unique features. On radiotherapy planning CT data of 57 patients, (14 symptomatic, 43 asymptomatic), a Support Vector Machine (SVM) obtained an area under the receiver operator curve (AUROC) of 0.873 with sensitivity, specificity and accuracy of 92%, 72% and 87% respectively. Furthermore, it was demonstrated that a Decision Tree classifier was capable of a similar level of performance using sub-regions of the lung volume. The second application is related to prostate cancer identification. T2 MRI scans are used in the diagnosis of prostate cancer and in the identification of the primary cancer within the prostate gland. The manual identification of the cancer relies on the assessment of multiple scans and the integration of clinical information by a clinician. This requires considerable experience and time. As MRI becomes more integrated within the radiotherapy work flow and as adaptive radiotherapy (where the treatment plan is modified based on multi-modality image information acquired during or between RT fractions) develops it is timely to develop automatic segmentation techniques for reliably identifying cancerous regions. In this work a number of texture features were coupled with a supervised learning model for the automatic segmentation of the main cancerous focus in the prostate - the focal lesion. A mean AUROC of 0.713 was demonstrated with 10-fold stratified cross validation strategy on an aggregate data set. On a leave one case out basis a mean AUROC of 0.60 was achieved which resulted in a mean DICE coefficient of 0.710. These results showed that is was possible to delineate the focal lesion in the majority (11) of the 14 cases used in the study.