A desconexão ázigo-portal e esplenectomia (DAPE) é a operação mais aceita e realizada em nosso meio para a profilaxia da recidiva hemorrágica por ruptura de varizes esofágicas ou gástricas em pacientes portadores de esquistossomose hepato-esplênica. Menores índices de ressangramento são obtidos através da associação da DAPE com escleroterapia ou ligadura elástica endoscópica das varizes esofágicas realizada no pós-operatório. Faltam trabalhos mostrando a evolução, a longo prazo, dos doentes esquistossomóticos submetidos a este tratamento. Este estudo retrospectivo tem como objetivo avaliar a evolução destes pacientes com tempo mínimo de seguimento de 5 anos. Foram analisados os prontuários dos pacientes operados no Serviço de Cirurgia de Fígado e Hipertensão Portal da Disciplina de Cirurgia do Aparelho Digestivo do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo, no período compreendido entre março de 1989 a março de 2001 e que foram acompanhados prospectivamente. Avaliamos dados clínicos, laboratoriais, endoscópicos e ultra-sonográficos de 97 pacientes com seguimento de 116,4 ± 46,7 meses. Nenhum paciente cursou com insuficiência hepática clínica ou laboratorial. Ocorreu, no pós-operatório tardio, correção da anemia, leucopenia e plaquetopenia, diminuição dos níveis de bilirrubinas total e indireta séricas e aumento do tempo de atividade da protrombina. Houve significativa redução do número e calibre das varizes esofágicas, assim como da presença de sinais de manchas vermelhas e de varizes gástricas. Houve aumento na freqüência de gastropatia congestiva, entretanto, sem repercussão clínica significativa. A recidiva hemorrágica xviii ocorreu em 24,7% dos pacientes, sendo em 14,6% quando considerada apenas por varizes esofágicas, gástricas ou duodenais. A probabilidade estimada de não ocorrer ressangramento em 20 anos é de 67,1%, sendo de 82,5% quando considerada recidiva por varizes. Em quatro pacientes a recidiva hemorrágica ocorreu por varizes, mesmo após o relato, em exame endoscópico prévio, de erradicação das varizes esofágicas. À avaliação ultrassonográfica observou-se redução do calibre da veia porta no pósoperatório tardio, em comparação ao pré-operatório. Concluímos que a desconexão ázigo-portal com esplenectomia, associada ao tratamento endoscópico de varizes esofágicas no pós-operatório, propicia bons resultados do ponto de vista clínico com baixa morbidade e mortalidade; permite melhora laboratorial da função hepática e correção do hiperesplenismo; determina a redução da incidência dos sinais endoscópicos preditivos de sangramento digestivo por hipertensão portal (varizes esofágicas de grosso calibre, sinais de manchas vermelhas e varizes de fundo gástrico), porém, a gastropatia congestiva é mais freqüente; permite adequada profilaxia da recidiva hemorrágica em 67% dos pacientes ao longo de 20 anos. A recidiva hemorrágica por varizes pode ocorrer mesmo após a erradicação das varizes esofágicas, tanto por recidiva de varizes como por varizes de outro sítio (gástrica ou duodenal). Ocorre redução do calibre da veia porta no pós-operatório tardio, observado em exame ultrassonográfico em comparação ao pré-operatório. / Esophagogastric devascularization and splenectomy (EGDS) is nowadays the most performed operation for esophageal varices bleeding recurrence prophylaxis in hepatosplenic schistosomiasis. Lower rebleeding rates are obtained through the association of EGDS with postoperative endoscopic sclerotherapy or elastic bandage of esophageal varices, however, there is a lack of studies showing long term results. The objectives of this study were to evaluate retrospectively EGDS results in patients with at least five years of follow-up. Clinical, laboratorial, endoscopic and ultrasonographic data of 97 patients submitted to EGDS from March 1989 to March 2001 were analyzed. The mean follow-up was 116.4 months. There was no postoperative clinical or laboratorial hepatic insufficiency. In the late follow-up we observed normalization of preexisting anemia, leucopenia, thrombocytopenia, hyperbilirubinemia, and a prothrombin activity time increase. There was a significant esophageal varices caliber and number reduction, cherry red spots signs and gastric varices decrease. Congestive gastropathy was observed more frequently but without clinical importance. Bleeding recurrence occurred in 24.7% of the patients, however, in 14.6% when esophageal varices hemorrhage was considered. Estimated probability of rebleeding prophylaxis over 20 years is 67.1% and 82.5% when variceal recurrence was considered. Bleeding recurrence occurred in four patients even after endoscopic evaluation showing esophageal varices eradication. There was a significant portal vein caliber reduction on late ultrasound assessment, compared to preoperative. We concluded that the EGDS with postoperative endoscopic treatment leads to good clinical results with low morbidity and mortality; provides laboratorial liver function improve and xx hypersplenism correction; determines endoscopic predictive signs of portal hypertension digestive bleeding decrease (large esophageal varices, cherry red spots signs and gastric varices), however congestive gastropathy is more frequent; allows appropriate bleeding prophylaxis in 67% of the patients over 20 years. Variceal hemorrhagic recurrence may occur even when esophageal varices eradication is reached suggesting the need of an endoscopic surveillance even in this group of patients.

O Câncer de Esôfago é uma das neoplasias mais agressivas do trato gastrointestinal; é considerado como a nona neoplasia mais comum no mundo, além de ter altas taxas de mortalidade. A doença pode ser desencadeada por hábitos de vida, tais como etilismo e tabagismo, e também por fatores intrínsecos ao indivíduo. De todo modo, interfere significativamente em vários setores da vida do sujeito. Uma doença como o câncer pode conter uma tentativa simbólica para compensar uma atitude unilateral do indivíduo, ao relacionar-se com o ambiente e consigo mesmo. Acredita-se que conhecer aspectos do funcionamento e da dinâmica psíquica do sujeito pode contribuir para o foco do tratamento psicológico, de forma a propiciar intervenções psicológicas precoces e a auxiliar as demais equipes assistenciais nos cuidados e no modo de interação com o paciente. Entre elementos da estrutura psíquica do sujeito, buscou-se a tipologia psicológica dos referidos pacientes; para tanto, foi utilizado o Questionário de avaliação tipológica - QUATI (Zacharias, 2003). Além disso, valemo-nos de aspectos contidos na avaliação psicológica, rotina da instituição, com a finalidade de acessarmos as particularidades de cada um dos pacientes, especialmente no que se refere à reação ao diagnóstico e ao significado atribuído à doença. O estudo pode ser considerado como transversal, exploratório e descritivo. Foram incluídos 90 pacientes, derivados de análise estatística. Foram sujeitos da pesquisa pacientes maiores de 18 anos com diagnóstico de câncer de esôfago; com ensino fundamental completo; virgens de tratamento; atendidos em primeira consulta, no ambulatório de cirurgia do aparelho digestivo, do Instituto do Câncer do Estado de São Paulo (ICESP-HCFMUSP). Encontramos esses pacientes em sala de espera da consulta médica, onde foram informados sobre o objetivo da pesquisa; e, àqueles que aceitaram participar, foi aplicado o termo de consentimento livre e esclarecido, seguido da avaliação psicológica (rotina da instituição) e aplicação do instrumento. Os resultados indicaram que a maior parte da nossa amostra tem uma atitude introvertida. O tipo psicológico mais comum foi o introvertido sensação, com função auxiliar sentimento. A partir da avaliação psicológica, constatou-se uma variedade de mecanismos defensivos para minimização da angústia. A grande parte dos pacientes valeu-se de defesas adaptativas diante do processo de adoecimento. Conclui-se que, no atendimento com esses pacientes, torna-se necessária uma postura prática e objetiva dos profissionais de saúde, em que se busque fornecer orientações com base na realidade dos pacientes, assim como ter um modo de agir empático, com o estabelecimento de vínculo significativo, mas respeitando os limites dos pacientes, sobretudo ao que se refere à forma reservada e introspectiva de interagirem com o mundo e com as pessoas que os cercam / Esophageal cancer is one of the most aggressive neoplasms of the gastrointestinal tract, and can be triggered by life habits such as alcoholism and smoking and factors intrinsic to the individual. A disease such as cancer can hold a symbolic attempt to compensate for a onesided attitude of the individual. It is believed that knowing aspects of the psychic dynamics of the person, can contribute to the focus of a psychological treatment, in order to provide early psychological interventions, as well as assisting other attending teams in the care of and interaction with the patient. Among the psychic structure elements of a subject, we looked for the psychological typology of these patients. Typological Assessment Questionnaire was used to evaluate the psychological typology. In addition, we drew on aspects from within the psychological assessment, routine of the institution, in order to access the particularities of each one of the patients, especially concerning their reaction to the diagnosis and the meaning attributed to the disease. Ninety patients were included. They were over 18 years old, having completed middle-school education, diagnosed with esophageal cancer, and interviewed during their first medical appointment at the Digestive Surgery Clinic at the Cancer Institute of the State of São Paulo (ICESP-HCFMUSP). We met these patients in the waiting room, where they were informed of the purpose of the survey. Those who agreed to participate, were given the terms of free and informed consent, followed by a psychological evaluation and the application of the instrument used. The results indicated a predominant introverted attitude. The most common psychological type was the introverted feeling, with feeling as a secondary function. From this psychological evaluation, we found a variety of defensive mechanisms to minimize the anguish. Most of the patients made use of adaptive defenses when facing the illness process. Therefore, when attending these patients, health professionals must have a practical and objective posture, seeking to guide the patients according to their reality. In addition, they should be empathetic in their manner, establishing significant bond, but respecting the patients\' limits, especially when it comes to their reserved and introspective way of interacting with the world and the people around them

INTRODUÇÃO: Há relatos de ocorrência de displasia e adenocarcinoma esofágico em pacientes com esôfago de Barrett submetidos a tratamento cirúrgico com fundoplicatura bem sucedido, comprovado pela realização de pHmetria esofágica prolongada sem evidência de acidez em esôfago, o que sugere que pode ocorrer refluxo distal ao cateter de pHmetria, não detectado por esse método. Essa hipótese motivou o desenvolvimento de metodologia para avaliar a presença de refluxo 1cm acima da borda superior do esfíncter inferior do esôfago (EIE). OBJETIVO: Comparar a exposição ácida em 3 diferentes níveis: 5cm acima da borda superior EIE, 1cm acima da borda superior do EIE e em região intra-esfincteriana. CASUÍSTICA E MÉTODO: 11 pacientes com esôfago de Barrett, submetidos à fundoplicatura à Nissen para tratamento do refluxo gastroesofágico, sem sintomas de refluxo e com endoscopia e estudo radiográfico contrastado de esôfago sem sinais de recidiva, foram selecionados. Foram submetidos à manometria esofágica para avaliar a localização e a extensão do EIE. Realizou-se então pHmetria esofágica com 4 canais: canal A: 5cm acima da borda superior do EIE; canal B: 1cm acima; canal C: intraesfincteriano, na porção média entre a borda superior e inferior do EIE; canal D: em posição intragástrica. Avaliou-se o escore de DeMeester no canal A, para detectar refluxo patológico. Comparou-se o número de episódios de refluxo ácido, o número de episódios de refluxo prolongado e a fração de tempo com pH < 4,0 nos canais A e B. Comparou-se a fração de tempo de pH < 4,0 nos canais B e C. A fração de tempo com pH < 4,0 no canal D foi usada como parâmetro para não migração proximal do cateter. RESULTADOS: Houve maior número de episódios de refluxo e maior fração de tempo com pH < 4,0 no canal B do que no canal A, com significância estatística, sendo a mediana do tempo de exposição ácida menor que 1,5%. Por outro lado, houve menor fração de tempo com pH < 4,0 no canal B do que no canal C, sugerindo competência do esfíncter em conter o refluxo gastroesofágico. Um paciente apresentou refluxo patológico no canal A, apesar de não ter evidência até então de refluxo. Dois casos de adenocarcinoma esofágico foram diagnosticados nos pacientes do grupo estudado. CONCLUSÕES: A região 1cm acima da borda superior do EIE está mais exposta a acidez do que a região 5cm acima da borda superior do EIE, embora a exposição ácida seja em níveis bem reduzidos. A região 1cm acima da borda superior do EIE está menos exposta a acidez do que a região intraesfincteriana, demonstrando eficácia da fundoplicatura em conter o refluxo nesse nível / INTRODUCTION: There are reports of dysplasia and esophageal adenocarcinoma occurring in patients with Barrett´s esophagus after successful surgical treatment with fundoplication, confirmed with prolonged esophageal pH monitoring showing no acid in esophagus, suggesting that there might be reflux distal to the catheter, not detected by this method. This hypothesis led to the development of methodology to evaluate the occurrence of reflux 1cm above the upper border of the lower esophageal sphincter (LES). OBJECTIVE: Compare the acid exposition in three different levels: 5cm above the upper border of the LES, 1cm above the upper border of the LES and in the intrasphincteric region. CASUISTIC AND METHODS: 11 patients with Barrett´s esophagus, submitted to Nissen fundoplication as treatment for gastroesophageal reflux, with no symptoms of reflux and with endoscopy and contrasted esophageal radiograph, were selected. The patients underwent esophageal manometry to evaluate the location and extension of the LES. After that, they underwent a 4-channel pH monitoring: channel A: 5cm above the upper border of the LES; channel B: 1cm above the upper border of the LES; channel C: intrasphincteric, in the mid-portion between the upper and lower border of the LES; channel D: intragastric. The DeMeester score was assessed in channel A, to detect pathologic reflux. The number of reflux episodes, the number of prolonged episodes and the fraction of time with pH < 4,0 were compared in channels A and B. The fraction of time with pH < 4,0 was compared in channels B and C. The fraction of time with pH < 4,0 in channel D was used as a parameter to ensure no proximal migration of the catheter. RESULTS: There were more episodes of reflux and a higher fraction of time with pH < 4,0 in channel B than in channel A, with statistical significance. On the other hand, there was lesser fraction of time with pH < 4,0 in channel B than in channel C, suggesting competence of the sphincter in preventing gastroesophageal reflux. One patient presented pathologic reflux in channel A, besides no other evidence of that. Two cases of esophageal adenocarcinoma were diagnosed in the studied patients. CONCLUSIONS: the region 1cm above the upper border of the LES is more exposed to acid than the region 5cm above the upper border of the LES, although this exposure occurred in reduced levels. The region 1cm above the upper border of the LES is less exposed to acid than the intrasphincteric region

Nas operações de cardiomiotomia para tratamento do megaesôfago ocorre recidiva dos sintomas em até 15% dos pacientes, sendo que alguns necessitam reoperação. Foram estudados de forma retrospectiva, 20 pacientes com megaesôfago previamente tratados por cardiomiotomia e submetidos a reoperação, por cardioplastia com gastrectomia parcial em Y-de-Roux, conforme técnica proposta por Serra Dória et al (1970). A causa de recidiva dos sintomas foi esofagite de refluxo em nove (45,0%), miotomia incompleta em um (5,0%), cicatrização da miotomia em cinco (25,0%) e presença de megaesôfago avançado em cinco (25,0%). Foram analisadas as complicações intra e pósoperatórias. Os pacientes foram avaliados sob o ponto de vista clínico (disfagia, regurgitação, pirose e variação de peso), radiológico e endoscópico, no pré e no pós-operatório imediato e tardio. Cinco (25,0%) doentes apresentaram complicações no pós-operatório imediato. Não houve mortalidade. Todos os doentes melhoraram da disfagia, com quase total desaparecimento da regurgitação e pirose. Houve manutenção ou aumento de peso em 64,7% dos pacientes. O estudo radiológico mostrou diminuição do calibre do esôfago em 53,0% e manutenção nos demais. Não houve aumento do calibre em nenhum dos pacientes. No exame endoscópico realizado no pós-operatório tardio em 17 pacientes, observou-se que seis entre nove que apresentavam esofagite de refluxo, melhoraram; dois entre oito, que apresentavam esôfago normal no préoperatório, desenvolveram esofagite. Concluiu-se que a operação de Serra Dória para tratamento do megaesôfago, operado por cardiomiotomia com recidiva dos sintomas apresentou baixa morbidade e ausência de mortalidade. Permitiu expressivo alívio dos sintomas e diminuição do calibre do esôfago em vários doentes. Possibilitou também melhora da esofagite de refluxo, havendo, entretanto, a possibilidade de sua manutenção, bem como do seu aparecimento em doentes que não a apresentavam / After cardiomyotomy for the treatment of megaesophagus, recurrence of symptoms occur in up to 15% of the patients, but only some require a reoperation. Twenty patients with megaesophagus were retrospectively studied. They were previously treated by cardiomyotomy, and underwent to reoperation through cardioplasty with Roux-en-Y partial gastrectomy, according to the technique proposed by Serra Dória et al (1970). The etiology of symptoms recurrence was reflux esophagitis in nine (45.0%) patients, incomplete myotomy in one (5.0%), healing of the myotomy in five (25.0%) and end staging megaesophagus in five (25.0%). Intra and postoperative complications were analyzed. The patients were studied by clinical (dysphagia, regurgitation, heartburn and weight gain), radiological and endoscopic evaluation, in the preand postoperative period. Five (25.0%) patients had complications in the immediate postoperative period. No deaths were observed. Dysphagia improved in all the patients. Regurgitation and heartburn almost disappeared in the whole group. Weight was maintained or increased in 64,7% of the patients. Radiological studies showed a decrease in the caliber of the esophagus in 53.0%, while the remaining patients maintained the pre-operative diameter. Endoscopy, performed during the late postoperative period in 17 patients, showed that six among the nine with reflux esophagitis improved; two among the eight with a normal esophagus during the preoperative period, developed esophagitis. It was concluded that the Serra Dória procedure for the treatment of megaesophagus in patients who had already undergone a cardiomyotomy and whose symptoms recurred, presented a low morbidity and no mortality. It afforded a significant relief of symptoms with a decrease of the caliber of the esophagus in several patients. The patients also improved with regards to reflux esophagitis. In some cases reflux was still present after surgery. Others with normal esophagus in the pre operative period developed esophagitis

Embora esteja comprovada a superioridade da ligadura elástica sobre a esclerose endoscópica na profilaxia secundária da hemorragia varicosa, ainda há discussão se esta vantagem também é observada no tratamento da fase aguda do sangramento. O presente estudo tem como objetivo comparar os resultados da ligadura elástica com a esclerose endoscópica em pacientes admitidos no Pronto-Socorro (PS) do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HCFMUSP) por hemorragia digestiva alta provocada por rotura de varizes de esôfago. Tratase de estudo unicêntrico, prospectivo, com alocação aleatória dos pacientes sem crossover. A fim de se detectar diferença de 20% entre a capacidade de hemostasia de cada um dos métodos, cada grupo deveria ser constituído por 260 pacientes, considerando poder estatístico de 80% e nível de significância inferior a 5%. Após diagnóstico endoscópico de rotura de varizes de esôfago, foi realizado sorteio para inclusão dos pacientes em dois grupos: esclerose endoscópica (EE) x ligadura elástica (LE). A esclerose foi realizada através da injeção intravasal de oleato de etanolamina a 3%, em alíquotas de 5ml, acima e abaixo do ponto de ruptura, respeitando o valor máximo por sessão de 20ml. No grupo LE, procurou-se ligar a variz sobre o ponto de ruptura. Se isto não foi possível, procurou-se ligar todo o tecido varicoso dos 3cm finais do esôfago. Foi utilizado o kit de ligadura MBL-6 e cateter VINF 23 (Cook, E. Tamusssino). As variáveis estudadas foram: a taxa de hemostasia inicial (até 5 dias), recidiva hemorrágica precoce (5 dias a 6 semanas), complicações e mortalidade. De maio de 2005 a maio de 2007, foram admitidos, no PS do HCFMUSP, 480 pacientes com hemorragia digestiva alta (HDA) provocada por hemorragia varicosa esofágica. Destes, 380 foram excluídos pelos seguintes motivos: mais de um tratamento prévio com ligadura ou esclerose (n=180), não randomização (n=85), uso de outra técnica hemostática como adesivo tissular de cianoacrilato (n=62) ou tratamento clínico incompleto no momento do exame endoscópico (n=53). Cem pacientes, 50 no grupo EE e 50 no LE foram incluídos no estudo. Destes, 72 eram homens e 28 mulheres, média de idade 52 anos. Os grupos se mostraram homogêneos quanto ao sexo, idade, Child-Pugh, hemoglobinemia à admissão, presença de choque hipovolêmico e calibre das varizes. Não foram encontradas diferenças estatisticamente significantes entre os grupos com relação ao controle inicial do sangramento (5 dias), ressangramento precoce (5 dias a 6 semanas), complicações e mortalidade (9 no grupo EE e 10 no grupo LE). Ao final de 6 semanas, 36 (80%) pacientes no grupo esclerose e 33 (77%) no grupo ligadura elástica estavam vivos e sem sinais de sangramento. Foi encontrada associação estatisticamente significante entre a classificação de Child-Pugh e mortalidade (p<0,001), que foi de 16% nos graus A ou B e 84% nos pacientes Child-Pugh C. Os resultados obtidos com esta casuística limitada sugerem que EE e a LE são igualmente eficazes no controle da hemorragia varicosa aguda. / Despite the superiority of banding over endoscopic sclerosis for secondary prophylaxis of variceal bleeding, there is still debate if this advantage is also observed for the acute bleeding setting. The study aims to compare band ligation (BL) with endoscopic sclerosis (SCL) in patients admitted to the emergency unit for rupture of esophageal varices. Prospective study carried out in a single center, with random allocation of the patients without crossover. In order to detect a 20% difference between the results of each method, each group should consist of 260 patients, considering an 80% statistical power and level of significance less than 5%. After an endoscopic diagnosis of rupture of esophageal varices, the patients were randomly allocated in two groups: SCL and BL. Sclerosis was performed by ethanolamine oleate intravascular injection, above and below the rupture point (maximum volume of 20 ml). In the BL group, banding was attempted at the point of rupture followed by ligation of the whole variceal tissue of the distal esophagus. Six-shooter® and VINF23® catheter (Cook, W. Salem, USA) were employed. Studied variables were initial failure in control bleeding (5 days), early rebleeding rates (5 days to 6 weeks), complications and mortality. From May 2005 to May 2007, 480 patients with an episode of variceal bleeding were admitted to the emergency room. From them, 380 were excluded because more than one previous treatment with SCL or BL (n=180), non-randomization (n=85), the use of another hemostatic technique such as cyanoacrylate tissular adhesive (n=62) and incomplete clinical treatment (n=53).One hundred patients, 50 in the SCL and 50 in the BL group were included in the study (72 male, 28 female, mean age 52 years). No differences between the groups were detected regarding gender, age, Child-Pugh status, the presence of shock at admission, mean hemoglobin levels and varices size. No statistically significant differences were found between the groups regarding control bleeding (5 days) and early rebleeding rates (5 days to 6 weeks), complications and mortality (9 in the SCL vs. 10 in the BL group). By the end of 6 weeks 36 (80%) patients in the SCL group and 33 (77%) in the EBL group were alive and free of bleeding. A statistically significant association was found between Child-Pugh status and mortality (p<0,001), which was 16% for A and B grades and 84% for grade C patients. The results obtained with this limited number of patients suggest that SCL and BL are equally efficient in the control of acute variceal bleeding.

Wang, Yajun. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 118-126). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.v / Table of Contents --- p.vi / List of Figures --- p.xi / List of Tables --- p.xii / Abbreviations and Symbols --- p.xiii / List of Publications and Sequence Submissions during the Study --- p.xv / Chapter Chapter One: --- General Introduction --- p.1 / Chapter Chapter Two: --- Literature Review --- p.8 / Chapter 2.1 --- DNA methylation --- p.8 / Chapter 2.1.1 --- Epigenetic changes --- p.8 / Chapter 2.1.2 --- Differential methylation pattern in normal and tumor cells --- p.10 / Chapter 2.2 --- TSGs --- p.13 / Chapter 2.2.1 --- "Cancer initiation, progression and cancer genes" --- p.13 / Chapter 2.2.2 --- TSGs could be inactivated through promoter hypermethylation --- p.14 / Chapter 2.3 --- NPC --- p.17 / Chapter 2.3.1 --- Epidemiology ofNPC --- p.18 / Chapter 2.3.2 --- Molecular genetic and epigenetic studies ofNPC --- p.19 / Chapter 2.3.3 --- NPC and chromosome 11q --- p.21 / Chapter 2.4 --- ESCC --- p.21 / Chapter 2.4.1 --- Epidemiology of ESCC --- p.22 / Chapter 2.4.2 --- Genetic and epigenetic studies of ESCC --- p.23 / Chapter 2.4.3 --- ESCC and chromosome 11q --- p.24 / Chapter 2.5 --- Chromosome 11q and other carcinomas --- p.24 / Chapter 2.5.1 --- Breast cancer --- p.24 / Chapter 2.5.2 --- Ovarian cancer --- p.25 / Chapter 2.5.3 --- Neuroblastoma --- p.26 / Chapter 2.5.4 --- Melanoma --- p.27 / Chapter 2.5.5 --- Multiple myeloma --- p.27 / Chapter 2.5.6 --- Lung Cancer --- p.27 / Chapter 2.6 --- Important candidate genes located at the project study 1 lq region --- p.28 / Chapter 2.6.1 --- ETS1 --- p.28 / Chapter 2.6.2 --- FLI1 --- p.29 / Chapter 2.6.3 --- P53AIP1 --- p.30 / Chapter 2.6.4 --- RICS --- p.30 / Chapter 2.6.5 --- BARX2 --- p.30 / Chapter 2.6.6 --- ST14 --- p.32 / Chapter 2.6.7 --- ADAMTS8 --- p.33 / Chapter 2.6.8 --- ADAMTS15 --- p.35 / Chapter 2.6.9 --- HNT --- p.36 / Chapter 2.6.10 --- OPCML --- p.36 / Chapter Chapter Three: --- Materials and Methods --- p.37 / Chapter 3.1 --- Cell lines and primary tumor samples --- p.37 / Chapter 3.2 --- Cell line demethylation treatment --- p.38 / Chapter 3.3 --- DNA and RNA extraction from cell lines and tissues --- p.39 / Chapter 3.4 --- Semiquantitative RT-PCR --- p.41 / Chapter 3.5 --- DNA bisulfite treatment --- p.42 / Chapter 3.6 --- Promoter analysis and identification of 5' CpG islands of target genes --- p.45 / Chapter 3.7 --- Methylation-Specific PCR (MSP) --- p.45 / Chapter 3.8 --- Bisulfite Genomic Sequencing (BGS) --- p.46 / Chapter 3.8.1 --- BGS PCR reaction --- p.46 / Chapter 3.8.2 --- TA cloning of the PCR products into the sequencing vector --- p.47 / Chapter 3.8.3 --- Plasmid mini-preparation on 96-well plate --- p.48 / Chapter 3.8.4 --- Plasmid sequencing --- p.49 / Chapter 3.9 --- Homozygous deletion detection --- p.50 / Chapter 3.10 --- Construction of expression plasmids --- p.51 / Chapter 3.10.1 --- The strategy of full length cDNA cloning --- p.51 / Chapter 3.10.2 --- Obtaining of full length covered cDNA by cloning PCR --- p.53 / Chapter 3.10.3 --- Ligation and transformation --- p.54 / Chapter 3.10.4 --- Mini preparation of plasmid in Eppendorf tubes --- p.54 / Chapter 3.10.5 --- Verification of correct inserts in the plasmid --- p.55 / Chapter 3.10.6 --- Subcloning --- p.55 / Chapter 3.10.7 --- Bacteria storage --- p.57 / Chapter 3.11 --- Colony formation assays (CFA) --- p.57 / Chapter 3.11.1 --- Midiprep of the transfection grade plasmid --- p.57 / Chapter 3.11.2 --- Transfection --- p.58 / Chapter 3.11.3 --- Selection of the transfected cells with G418 --- p.59 / Chapter 3.11.4 --- Colony staining --- p.60 / Chapter 3.12 --- Statistical analysis --- p.60 / Chapter Chapter Four: --- Results --- p.61 / Chapter 4.1 --- Narrow down the candidate genes for further study --- p.61 / Chapter 4.1.1 --- Define the study chromosome region --- p.61 / Chapter 4.1.2 --- Database search of all candidate genes --- p.61 / Chapter 4.1.3 --- Transcriptional expression analysis of the candidate genes --- p.63 / Chapter 4.1.4 --- Selection of the genes with tumor specific expression downregulation for further intensive study --- p.64 / Chapter 4.2 --- Further characterization of ADAMTS8 --- p.69 / Chapter 4.2.1 --- Tissue transcriptional expression panel --- p.69 / Chapter 4.2.2 --- Semiquantitative RT-PCR results in tumor cell lines --- p.70 / Chapter 4.2.3 --- Promoter CpG island identification and promoter methylation study --- p.70 / Chapter 4.2.4 --- Transcription reactivation by demethylation treatment --- p.72 / Chapter 4.2.5 --- High resolution promoter methylation analysis by BGS --- p.72 / Chapter 4.2.6 --- Detection of homozygous deletion --- p.73 / Chapter 4.2.7 --- Analysis of ADAMTS8 promoter methylation in clinical samples --- p.74 / Chapter 4.2.8 --- ADAMTS8 full length cDNA cloning --- p.74 / Chapter 4.2.9 --- Colony formation assay --- p.75 / Chapter 4.3 --- Further characterization of HNT --- p.80 / Chapter 4.3.1 --- Tissue transcriptional expression panel --- p.80 / Chapter 4.3.2 --- Semiquantitative RT-PCR results in tumor cell lines --- p.80 / Chapter 4.3.3 --- Promoter CpG island identification and promoter methylation study --- p.81 / Chapter 4.3.4 --- Transcription reactivation by demethylation treatment --- p.82 / Chapter 4.3.5 --- HNT full length cDNA cloning --- p.82 / Chapter 4.4 --- Further characterization of BARX2 --- p.87 / Chapter 4.4.1 --- Tissue transcriptional expression panel --- p.87 / Chapter 4.4.2 --- Semiquantitative RT-PCR results in tumor cell lines --- p.87 / Chapter 4.4.3 --- Promoter CpG island identification and promoter methylation study --- p.88 / Chapter 4.4.4 --- Transcription reactivation by demethylation treatment --- p.89 / Chapter 4.4.5 --- BARX2 full length cDNA cloning --- p.89 / Chapter 4.5 --- Further study of other downregulated genes --- p.92 / Chapter 4.5.1 --- FLII --- p.92 / Chapter 4.5.2 --- ADAMTS15 --- p.94 / Chapter 4.5.3 --- P53AIP1 --- p.97 / Chapter Chapter Five: --- Discussion --- p.100 / Reference List --- p.118 / Appendix I: Reagents Preparation Recipe --- p.127 / Appendix II: PCR Primers for cDNA Cloning --- p.129

Orientador: Fernando Gomes Romeiro / Resumo: Introdução: O tratamento das alterações ósseas em portadores de cirrose ainda não está bem definido, fazendo com que vários pacientes permaneçam sem tratamento e venham a ter complicações graves como fraturas e dores crônicas. Objetivos: Avaliar a segurança e a eficácia do tratamento da osteoporose e osteopenia em indivíduos com cirrose e varizes de esôfago de baixo risco de sangramento. Materiais e métodos: indivíduos com osteoporose receberam risedronato associado a suplementação de cálcio e vitamina D, e os com osteopenia receberam somente a suplementação nutricional. A análise sobre a segurança foi realizada por endoscopias pré-tratamento, após 6 e 12 meses, bem como por consultas após um ano para avaliar se houve eventos adversos. Esses resultados foram comparados entre o grupo osteoporose e o grupo osteopenia pelo teste exato de Fisher. A análise sobre a eficácia foi realizada por meio da Dual Energy X-Ray Absorptiometry (DXA), comparando valores pré-tratamento e após 2 anos em cada indivíduo através da estimativa do intervalo de confiança e pelo teste t de Student. Resultados: Os achados endoscópicos e eventos adversos foram semelhantes entre os grupos. No grupo osteoporose houve aumento do T-escore da coluna lombar (p<0,001), sem sinais de perda óssea no colo femoral. O grupo osteopenia não teve melhora óssea vertebral e sofreu redução do T-escore do colo do fêmur (p= 0,019). Conclusão: Os tratamentos foram seguros, mas somente o com risedronato proporcionou melhora ó... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction: The treatment of bone loss in patients with cirrhosis is not well defined, so that many patients do not receive effective therapy and develop complications such as fractures and chronic pain. Aim: To evaluate the safety and the efficacy of osteoporosis and osteopenia treatment in patients with liver cirrhosis and low-risk esophageal varices. Materials and methods: The osteoporosis treatment consisted of risedronate associated with calcium and vitamin D supplementation, while the osteopenia treatment comprised only the nutritional supplementation. The safety assessment was done by endoscopic exams before and during the treatment (6 and 12 months), as well as by outpatient appointments carried out 1 year after starting the treatment. These results were compared between the osteoporosis and osteopenia groups through the Fisher exact test. The efficacy analysis was measured through the T-score assessed by Dual Energy X-Ray Absorptiometry (DXA), which was performed before and at 2 years of treatment by calculating the 95% confidence interval and applying the t-Student test. Results: The endoscopic findings and the adverse events were similar between the groups. The osteoporosis group had a significant increase in the lumbar spine T-score (p <0.001), with no bone loss in the femoral neck. The osteopenia group had no improvement in vertebral bones and suffered a clear reduction in the femoral neck T-score (p = 0.019). Conclusion: The treatments were safe but only the o... (Complete abstract click electronic access below) / Doutor

O Objetivo desse trabalho é avaliar a Dinâmica Gástrica e o Trânsito Oro-Esofágico em imagens cintilográficas por meio de uma técnica de fluxo óptico chamada Técnica Aprimorada de Demons. Essa técnica permite obter a velocidade dos pixels, também chamada Força de Deformação, em imagens sequenciais aos pares por meio de 3 características das imagens: o gradiente, a intensidade dos pixels e a informação ortogonal ao gradiente. Para alcançar o nosso objetivo, foram utilizadas imagens cintilográficas obtidas no Banco de Imagens do Hospital das Clínicas da USP de Ribeirão Preto. Essas imagens foram adquiridas segundo 2 protocolos: Trânsito Oro-Esofágico e Enchimento Gástrico. Sob o protocolo de Trânsito Oro-Esofágico, foram selecionadas imagens referentes a 9 voluntários saudáveis. Esses, em um período de jejum mínimo de 8 horas, fizeram 10 deglutições de 5 ml de solução fisiológica sendo as deglutições de número 1, 4, 7 e 10 feitas com a solução fisiológica \"marcada\'\' com aproximadamente 26 MBq (cerca de 700 µCi) de 99mTecnécio ligado a moléculas de fitato e adquiridas, pela gamma câmara, como um conjunto de 240 imagens por deglutição. As imagens adquiridas no protocolo de Trânsito Oro-Esofágico foram corrigidas e corregistradas. Os campos de velocidade obtidos foram correlacionados com uma técnica capaz de obter os tempos de trânsito do bolus nas diferentes porções do esôfago. Foi obtida uma alta correlação inversa entre as velocidades e o tempo de trânsito ao longo de todo o esôfago, como esperado. A média das velocidades obtidas no esôfago proximal se aproximou consideravelmente da velocidade da onda peristáltica primária descrita na literatura. Sob o protocolo de Enchimento Gástrico, foram selecionadas imagens referentes a 13 voluntários saudáveis. Os voluntários, em um período de jejum mínimo de 8 horas, ingeriram uma refeição teste líquida \"marcada\'\' com aproximadamente 72 MBq (cerca de 2 mCi) de 99mTecnécio ligado a moléculas de fitato enquanto ocorria a aquisição de um conjunto de 330 imagens pela gamma câmara. As imagens adquiridas no protocolo de Enchimento Gástrico também foram corrigidas e corregistradas. Os campos de velocidades obtidos foram somados para cada par de imagens adquiridas e foi analisada a frequência dominante do módulos desses campos somados no tempo de aquisição por meio da Trânsformada Rápida de Fourier (FFT), com filtro Butterworth bidirecional, em janelas de 4 minutos. Foi obtida a média das frequências dominantes por voluntário e, posteriormente, uma média e um desvio padrão para a amostra de voluntários. Foi feito um teste t de student que revelou que a média amostral da frequência dominante do estômago proximal é condizente com a frequência de contração conhecida na literatura para essa região (1 contração por minuto). Ao se estudar a região distal, foi observado também uma predominância de janelas com frequências próximas a conhecida para contrações do estômago distal (3 contrações por minuto) na maioria dos voluntários. As contrações no estômago também foram analisadas visualmente por meio do divergente do campo de velocidades e do perímetro do estômago nas imagens adquiridas. / The goal of this project is to evaluate the gastric dinamic and the Oro-esophageal transit in scintigraphic images trough an optical flow tecnique called Improved Demons Tecnique. This tecnique provides the velocity of the pixels (also called, Deformation Force - DF) in sequencial images by pairs trough 3 image features: gradient, intensity of the pixels and orthogonal gradient information. To achieve our goal, scintigraphic images from Hospital das Clínicas of Ribeirão Preto database were used. This images were acquired trough 2 protocols: Oro-Esophageal Transit and Gastric Filling. At the Oro-Esophageal Transit protocol, images from 9 healthy volunteers were chosen. The volunteers, under a fasting of at least 8 hours, made ten swallows of 5 ml saline. The swallows number 1, 4, 7 and 10 were made with the saline labelled with about 26 MBq (about 700 µCi) 99mTc-phytate and acquired, by gamma-camera, as a set of 240 images per swallowing. The images acquired under the protocol Oro-Esophageal Transit were corrected and registered. The velocity fields obtained were correlated to a tecnique capable of obtain the relative transit times of the bolus in each region of the oesophagus. A high inverse correlation were obtained between the velocities and the times of transit in the oesophagus as a whole, as expected. The mean of the velocities obtained in the proximal oesophagus approached considerably of the velocity of primary peristaltic pump described in literature. At the Gastric filling protocol, images from 13 healthy volunteers were chosen. The volunteers, under a fasting of at least 8 hours, ingested a liquid meal labeled with about 72 MBq (about 2 mCi) 99mTc-phytate while a set of 330 images were acquired by gamma-camera. This images were also corrected and registered. The velocity fields were summed for each pair of images acquired and the frequency of the magnitude of this summed field, in the time of acquisition, were analyzed by Fast Fourier Transform, with a bidirectional Butterworth filter, in sliding windows of 4 minutes. A mean of the frequencies per volunteer were obtained and, after, a mean and standard deviation for the sample of volunteers. It was made a t student test that showed that the sample mean of the frequency of the proximal estomach is consistent with the known frequency of the contraction for this region (1 contraction per minute). We also studied the distal region. It was observed a predominance of slider windows with frequencies close to the frequency known for contractions of distal stomach (3 contractions per minute) in most of the volunteers. The stomach contractions were also analyzed visually through the divergent of the velocity fields and the perimeter of the stomach in the acquired images.

Yu, Le. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 171-198). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese.

In brief, mRNA expression profiling of candidate genes in each locus was performed using semi-quantitative RT-PCR in a panel of ESCC and NPC cell lines, normal tissues and immortalized epithelial cell lines. Genes downregulated in cancer cells but with high expression in normal tissues and immortalized epithelial cells were subjected to promoter methylation analysis using methylation-specific PCR (MSP), bisulfite genomic sequencing (BGS) and pharmacological demethylation treatment. Genes with tumor-specific downregulation and methylation were further selected as candidates and their tumor suppressive roles were verified via functional studies. / In conclusion, RAB39 and WDRX, epigenetically silenced in multiple cancer cell lines, were identified as novel TSG candidates in this study. Meanwhile, the tumor suppressive functions of ADAMTS8 were further validated, proving the efficiency of this integrative approach. Further study on these novel TSG candidates may help to elucidate the detailed molecular mechanisms for ESCC and NPC, and provide novel therapeutic targets and biomarkers. / In this study, RAB39 and WDRX were identified as candidate TSGs in 11q22.3 and 15q21.3, respectively. Both genes were broadly expressed in normal tissues and immortalized epithelial cell lines, but significantly downregulated and methylated in multiple cancer cell lines. Demethylation treatment with 5-Aza-2'-deoxycytidine restored their mRNA expression, indicating that CpG methylation directly contributed to their transcriptional inactivation. Methylation of RAB39 and WDRX was detected in primary ESCC and NPC, but rarely observed in normal tissues, implicating that their tumor-specific methylation might be used as biomarkers. Ectopic expression of both genes significantly inhibited the clonogenicity of multiple cancer cell lines, supporting their potential roles as functional TSGs. Moreover, WDRX repressed WNT/beta-catenin signaling, underscoring a possible anti-tumorigenic mechanism for it. In addition, ADAMTS8 was revealed to inhibit clonogenicity of NPC and ESCC cell lines, acting as a negative modulator for ERK pathway and a potential pro-apoptotic metalloprotease. / Inactivation of tumor suppressor genes (TSGs) contributes to the genesis of cancers including esophageal squamous cell carcinoma (ESCC) and nasopharyngeal carcinoma (NPC), two prevalent causes of death in Hong Kong. Apart from genetic abnormalities, epigenetic disruptions including CpG methylation represent another major mechanism for TSG inactivation. Promoter methylation of multiple TSGs was detected in different cancer types, suggesting that it could be utilized as therapeutic target or biomarker for disease diagnosis and prognosis. / TSGs are often located at frequently deleted chromosomal regions and subjected to tumor-specific methylation, making it possible to use an integrative epigenetic and genomic approach combining array comparative genomic hybridization (aCGH) with epigenetic profiling to screen for novel TSGs. Previous aCGH revealed that several loci in 11822.3, 15q14, 15q21.1 and 15q21.3 underwent frequent copy number loss in ESCC cell lines. Loss of heterozygosity (LOH) of these regions was also reported in other cancers, indicating that TSGs might reside within them. The aim of this study was thus to identify the candidate TSGs in these loci and study their anti-tumorigenic roles. In addition, the tumor suppressive function of ADAMTS8, a silenced 11q25 candidate TSG previously identified in our lab via this approach, was also studied. / Li, Jisheng. / Adviser: Qian Tao. / Source: Dissertation Abstracts International, Volume: 72-04, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 136-159). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.