RESUMO Contexto: Aproximadamente 71 milhões de pessoas estão infectadas pelo vírus da hepatite C em todo o mundo. Um número significativo desses indivíduos desenvolverá cirrose hepática e/ou carcinoma hepatocelular. Além do fígado, há evidências científicas que associam doenças cardiovasculares e hepatite C crônica; no entanto, os mecanismos biológicos implicados na ocorrência dessas condições ainda não foram completamente esclarecidos. Objetivo: Avaliar a associação entre histologia hepática, comorbidades clínicas e perfil lipídico em pacientes com hepatite C crônica. Investigar associações entre histologia hepática e parâmetros demográficos, nutricionais, bioquímicos e virológicos. Métodos: Oitenta e cinco pacientes com hepatite C crônica foram prospectivamente submetidos à biópsia hepática. Biópsias hepáticas foram obtidas de cada paciente por via percutânea com agulha de Menghini. A fibrose foi avaliada de acordo com o sistema de pontuação METAVIR, como segue: F0, sem fibrose; F1, expansão portal fibrosa; F2, alargamento portal fibroso com poucos septos; F3, fibrose em ponte com distorção arquitetônica; e F4, cirrose hepática. A atividade foi classificada com base no grau de infiltração de linfócitos e necrose de hepatócitos, de A0 a A3. O diagnóstico da doença hepática foi baseado em métodos clínicos, bioquímicos, histológicos e radiológicos. Os dados foram analisados por modelos de regressão logística. Resultados: Neste estudo transversal, realizado em um ambulatório do hospital universitário, foram incluídos 85 pacientes que tinham média de idade de 57,2±10,7 anos, sendo 45 (52,9%) do sexo feminino. Havia 10 pacientes com cirrose. Os pacientes com METAVIR F3-F4 eram significativamente mais velhos (P=0,02) e tinham níveis mais elevados de ALT (P=0,0006), AST (P<0,0001), γ-GT (P=0,03) e bilirrubina (P=0,001) e, maior tempo de protrombina do que pacientes com escore F0-F2. Os níveis de albumina (P=0,01) foram significativamente mais baixos naqueles classificados como METAVIR F3-F4. Idade (OR=1,09; IC95%=1,02-1,16; P=0,02), esteatose (OR=4,03; IC95%=1,05-15,45; P=0,04) e HDL-C <60 mg/dL (OR=7,67; 95%IC=1,71-34,49; P=0,008) foram independentemente associados à fibrose. Hipertensão (OR=6,36; IC95%=1,31-30,85; P=0,02) e HDL-C <60 mg/dL (OR=9,85; IC95%=2,35-41,39; P=0,002) foram independentemente associados à atividade necroinflamatória. Hipertensão (OR=6,94; IC 95%=1,92-25,05; P=0,003) e HDL-C <60 mg/dL (OR=3,94; IC95%=1,27-12,3; P=0,02) foram associados à atividade inflamatória de interface. Os triglicerídeos (TG >150 mg/dL) permaneceram associados à atividade inflamatória lobular. Conclusão: Níveis de coleterol HDL <60 mg/dL foram independentemente associados à atividade necroinflamatória na hepatite C crônica. Pacientes com hipertensão têm risco aumentado de desenvolver atividade necroinflamatória. Contexto 7 mundo eou ou hepatocelular fígado entanto esclarecidos Objetivo demográficos nutricionais virológicos Métodos Menghini segue F0 F F1 fibrosa F2 septos F3 arquitetônica F4 hepatócitos A3 clínicos radiológicos logística Resultados transversal universitário 8 572107 57 2 57,2±10, anos 4 52,9% 529 52 9 (52,9% feminino 1 F3F4 FF F3-F P=0,02 P002 P 0 02 (P=0,02 P=0,0006, P00006 P=0,0006 , 0006 (P=0,0006) P<0,0001, P00001 P<0,0001 0001 (P<0,0001) γGT γ GT P=0,03 P003 03 (P=0,03 P=0,001 P0001 001 (P=0,001 F0F2. F0F2 F2. F0-F2 P=0,01 P001 01 (P=0,01 F3F4. F4. OR=1,09 OR109 OR 09 (OR=1,09 IC95%=1,021,16 IC95102116 IC95%=1,02 1,16 IC95 16 IC95%=1,02-1,16 P=0,02, OR=4,03 OR403 (OR=4,03 IC95%=1,0515,45 IC951051545 IC95%=1,05 15,45 05 15 IC95%=1,05-15,45 P=0,04 P004 04 HDLC 60 <6 mgdL mg dL OR=7,67 OR767 67 (OR=7,67 95%IC=1,7134,49 95IC1713449 95%IC=1,71 34,49 95 34 49 95%IC=1,71-34,49 P=0,008 P0008 008 OR=6,36 OR636 6 36 (OR=6,36 IC95%=1,3130,85 IC951313085 IC95%=1,31 30,85 31 30 IC95%=1,31-30,85 OR=9,85 OR985 (OR=9,85 IC95%=2,3541,39 IC952354139 IC95%=2,35 41,39 35 41 39 IC95%=2,35-41,39 P=0,002 P0002 002 OR=6,94 OR694 94 (OR=6,94 95%=1,9225,05 951922505 95%=1,92 25,05 92 25 95%=1,92-25,05 P=0,003 P0003 003 OR=3,94 OR394 3 (OR=3,94 IC95%=1,2712,3 IC95127123 IC95%=1,27 12,3 27 12 IC95%=1,27-12,3 interface TG 150 >15 lobular Conclusão 57210 5 57,2±10 52,9 (52,9 F3F P=0,0 P00 (P=0,0 P0000 P=0,000 000 (P=0,0006 P<0,000 (P<0,0001 P=0,00 P000 00 (P=0,00 F0F F0-F OR=1,0 OR10 (OR=1,0 021 IC95%=1,021,1 IC9510211 IC95102 IC95%=1,0 116 1,1 IC9 IC95%=1,02-1,1 OR=4,0 OR40 (OR=4,0 0515 IC95%=1,0515,4 IC95105154 IC95105 1545 15,4 IC95%=1,05-15,4 < OR=7,6 OR76 (OR=7,6 7134 95%IC=1,7134,4 95IC171344 95IC171 95%IC=1,7 3449 34,4 95%IC=1,71-34,4 OR=6,3 OR63 (OR=6,3 3130 IC95%=1,3130,8 IC95131308 IC95131 IC95%=1,3 3085 30,8 IC95%=1,31-30,8 OR=9,8 OR98 (OR=9,8 3541 IC95%=2,3541,3 IC95235413 IC95235 IC95%=2,3 4139 41,3 IC95%=2,35-41,3 OR=6,9 OR69 (OR=6,9 9225 95%=1,9225,0 95192250 95192 95%=1,9 2505 25,0 95%=1,92-25,0 OR=3,9 OR39 (OR=3,9 2712 IC95%=1,2712, IC9512712 IC95127 IC95%=1,2 123 12, IC95%=1,27-12, >1 5721 57,2±1 52, (52, P=0, P0 (P=0, (P=0,000 P<0,00 (P<0,000 OR=1, OR1 (OR=1, IC95%=1,021, IC951021 IC9510 IC95%=1, 11 1, IC95%=1,02-1, OR=4, OR4 (OR=4, 051 IC95%=1,0515, IC9510515 154 15, IC95%=1,05-15, OR=7, OR7 (OR=7, 713 95%IC=1,7134, 95IC17134 95IC17 95%IC=1, 344 34, 95%IC=1,71-34, OR=6, OR6 (OR=6, 313 IC95%=1,3130, IC9513130 IC9513 308 30, IC95%=1,31-30, OR=9, OR9 (OR=9, 354 IC95%=2,3541, IC9523541 IC9523 IC95%=2, 413 41, IC95%=2,35-41, 922 95%=1,9225, 9519225 9519 95%=1, 250 25, 95%=1,92-25, OR=3, OR3 (OR=3, 271 IC95%=1,2712 IC951271 IC9512 IC95%=1,27-12 > 572 57,2± (52 P=0 (P=0 P<0,0 (P<0,00 OR=1 (OR=1 IC95%=1,021 IC951 IC95%=1 IC95%=1,02-1 OR=4 (OR=4 IC95%=1,0515 IC951051 IC95%=1,05-15 OR=7 (OR=7 95%IC=1,7134 95IC1713 95IC1 95%IC=1 95%IC=1,71-34 OR=6 (OR=6 IC95%=1,3130 IC951313 IC95%=1,31-30 OR=9 (OR=9 IC95%=2,3541 IC952354 IC952 IC95%=2 IC95%=2,35-41 95%=1,9225 951922 951 95%=1 95%=1,92-25 OR=3 (OR=3 IC95%=1,271 IC95%=1,27-1 57,2 (5 P= (P= P<0, (P<0,0 OR= (OR= IC95%= IC95%=1,02- IC95%=1,051 IC95%=1,05-1 95%IC=1,713 95IC 95%IC= 95%IC=1,71-3 IC95%=1,313 IC95%=1,31-3 IC95%=2,354 IC95%=2,35-4 95%=1,922 95%= 95%=1,92-2 IC95%=1,27- 57, ( (P P<0 (P<0, (OR IC95% IC95%=1,05- 95%IC 95%IC=1,71- IC95%=1,31- IC95%=2,35- 95% 95%=1,92- P< (P<0 (P<

ABSTRACT Background: Approximately 71 million people are chronically infected with hepatitis C virus (HCV) worldwide. A significant number of these individuals will develop liver cirrhosis and/or hepatocellular carcinoma. Beyond the liver, there is a sizeable body of scientific evidence linking cardiovascular disease and chronic hepatitis C (CHC); however, the biological mechanisms behind the concurrence of these conditions have not been completely clarified yet. Objective: To evaluate associations between hepatic histology, clinical comorbidities and lipid profile in patients with CHC. To investigate associations between liver histology and demographic, nutritional, biochemical and virological parameters. Methods: Eight-five patients with CHC prospectively underwent hepatic biopsy. Liver fragments were obtained from each patient by percutaneous route using a Menghini needle. Fibrosis was evaluated according to the METAVIR scoring system, as follows: F0, no fibrosis; F1, fibrous portal expansion; F2, fibrous portal widening with few septa; F3, bridging fibrosis with architectural distortion; and F4, liver cirrhosis. The activity was classified based on the degree of lymphocyte infiltration and hepatocyte necrosis, from A0 to A3. The diagnosis of liver disease was based on clinical, biochemical, histological, and radiological methods. The data were analyzed by logistic regression models. Results: This cross-sectional study included 85 outpatients followed at the tertiary care ambulatory centre with a mean age of 57.2±10.7 years and 45 (52.9%) were females. There were 10 patients with cirrhosis. Patients with a METAVIR F3-F4 were significantly older (P=0.02) and had higher levels of ALT (P=0.0006), AST (P<0.0001), γ-GT (P=0.03) and bilirubin (P=0.001) and higher prothrombin time than patients with F0-F2 score. Albumin levels (P=0.01) were significantly lower in METAVIR F3-F4. Age (OR=1.09; 95%CI=1.02-1.16; P=0.02), steatosis (OR=4.03; 95%CI=1.05-15.45; P=0.04) and high-density lipoprotein cholesterol (HDL-C) <60 mg/dL (OR=7.67; 95%CI=1.71-34.49; P=0.008) were independently associated with fibrosis. Hypertension (OR=6.36; 95%CI=1.31-30.85; P=0.02) and HDL-C <60 mg/dL (OR=9.85; 95%CI=2.35-41.39; P=0.002) were independently associated with necroinflammatory activity. Hypertension (OR=6.94; 95%CI=1.92-25.05; P=0.003) and HDL-C <60 mg/dL (OR=3.94; 95%CI=1.27-12.3; P=0.02) were associated with interface inflammatory activity. Triglycerides (TG ≥150 mg/dL) remained associated with lobular inflammatory activity. Conclusion: cholesterol levels <60 mg/dL were independently associated with necroinflammatory activity in chronic hepatitis C. Patients with hypertension are at an increased risk of developing necroinflammatory activity. Background 7 HCV (HCV worldwide andor or carcinoma (CHC) however yet Objective demographic nutritional parameters Methods Eightfive Eight five biopsy needle system follows F0 F F1 expansion F2 septa F3 distortion F4 necrosis A3 histological methods models Results crosssectional cross sectional 8 572107 57 2 57.2±10. 4 52.9% 529 52 9 (52.9% females 1 F3F4 FF F3-F P=0.02 P002 P 0 02 (P=0.02 P=0.0006, P00006 P=0.0006 , 0006 (P=0.0006) P<0.0001, P00001 P<0.0001 0001 (P<0.0001) γGT γ GT P=0.03 P003 03 (P=0.03 P=0.001 P0001 001 (P=0.001 F0F2 F0-F score P=0.01 P001 01 (P=0.01 F3F4. F4. OR=1.09 OR109 OR 09 (OR=1.09 95%CI=1.021.16 95CI102116 CI 95%CI=1.02 1.16 95 16 95%CI=1.02-1.16 P=0.02, OR=4.03 OR403 (OR=4.03 95%CI=1.0515.45 95CI1051545 95%CI=1.05 15.45 05 15 95%CI=1.05-15.45 P=0.04 P004 04 highdensity high density HDLC HDL (HDL-C 60 <6 mgdL mg dL OR=7.67 OR767 67 (OR=7.67 95%CI=1.7134.49 95CI1713449 95%CI=1.71 34.49 34 49 95%CI=1.71-34.49 P=0.008 P0008 008 OR=6.36 OR636 6 36 (OR=6.36 95%CI=1.3130.85 95CI1313085 95%CI=1.31 30.85 31 30 95%CI=1.31-30.85 OR=9.85 OR985 (OR=9.85 95%CI=2.3541.39 95CI2354139 95%CI=2.35 41.39 35 41 39 95%CI=2.35-41.39 P=0.002 P0002 002 OR=6.94 OR694 94 (OR=6.94 95%CI=1.9225.05 95CI1922505 95%CI=1.92 25.05 92 25 95%CI=1.92-25.05 P=0.003 P0003 003 OR=3.94 OR394 3 (OR=3.94 95%CI=1.2712.3 95CI127123 95%CI=1.27 12.3 27 12 95%CI=1.27-12.3 TG 150 ≥15 Conclusion (CHC 57210 5 57.2±10 52.9 (52.9 F3F P=0.0 P00 (P=0.0 P0000 P=0.000 000 (P=0.0006 P<0.000 (P<0.0001 P=0.00 P000 00 (P=0.00 F0F OR=1.0 OR10 (OR=1.0 021 95%CI=1.021.1 95CI10211 95CI102 95%CI=1.0 116 1.1 95%CI=1.02-1.1 OR=4.0 OR40 (OR=4.0 0515 95%CI=1.0515.4 95CI105154 95CI105 1545 15.4 95%CI=1.05-15.4 < OR=7.6 OR76 (OR=7.6 7134 95%CI=1.7134.4 95CI171344 95CI171 95%CI=1.7 3449 34.4 95%CI=1.71-34.4 OR=6.3 OR63 (OR=6.3 3130 95%CI=1.3130.8 95CI131308 95CI131 95%CI=1.3 3085 30.8 95%CI=1.31-30.8 OR=9.8 OR98 (OR=9.8 3541 95%CI=2.3541.3 95CI235413 95CI235 95%CI=2.3 4139 41.3 95%CI=2.35-41.3 OR=6.9 OR69 (OR=6.9 9225 95%CI=1.9225.0 95CI192250 95CI192 95%CI=1.9 2505 25.0 95%CI=1.92-25.0 OR=3.9 OR39 (OR=3.9 2712 95%CI=1.2712. 95CI12712 95CI127 95%CI=1.2 123 12. 95%CI=1.27-12. ≥1 5721 57.2±1 52. (52. P=0. P0 (P=0. (P=0.000 P<0.00 (P<0.000 OR=1. OR1 (OR=1. 95%CI=1.021. 95CI1021 95CI10 95%CI=1. 11 1. 95%CI=1.02-1. OR=4. OR4 (OR=4. 051 95%CI=1.0515. 95CI10515 154 15. 95%CI=1.05-15. OR=7. OR7 (OR=7. 713 95%CI=1.7134. 95CI17134 95CI17 344 34. 95%CI=1.71-34. OR=6. OR6 (OR=6. 313 95%CI=1.3130. 95CI13130 95CI13 308 30. 95%CI=1.31-30. OR=9. OR9 (OR=9. 354 95%CI=2.3541. 95CI23541 95CI23 95%CI=2. 413 41. 95%CI=2.35-41. 922 95%CI=1.9225. 95CI19225 95CI19 250 25. 95%CI=1.92-25. OR=3. OR3 (OR=3. 271 95%CI=1.2712 95CI1271 95CI12 95%CI=1.27-12 ≥ 572 57.2± (52 P=0 (P=0 P<0.0 (P<0.00 OR=1 (OR=1 95%CI=1.021 95CI1 95%CI=1 95%CI=1.02-1 OR=4 (OR=4 95%CI=1.0515 95CI1051 95%CI=1.05-15 OR=7 (OR=7 95%CI=1.7134 95CI1713 95%CI=1.71-34 OR=6 (OR=6 95%CI=1.3130 95CI1313 95%CI=1.31-30 OR=9 (OR=9 95%CI=2.3541 95CI2354 95CI2 95%CI=2 95%CI=2.35-41 95%CI=1.9225 95CI1922 95%CI=1.92-25 OR=3 (OR=3 95%CI=1.271 95%CI=1.27-1 57.2 (5 P= (P= P<0. (P<0.0 OR= (OR= 95CI 95%CI= 95%CI=1.02- 95%CI=1.051 95%CI=1.05-1 95%CI=1.713 95%CI=1.71-3 95%CI=1.313 95%CI=1.31-3 95%CI=2.354 95%CI=2.35-4 95%CI=1.922 95%CI=1.92-2 95%CI=1.27- 57. ( (P P<0 (P<0. (OR 95%CI 95%CI=1.05- 95%CI=1.71- 95%CI=1.31- 95%CI=2.35- 95%CI=1.92- P< (P<0 (P<

ABSTRACT BACKGROUND: Occult hepatitis B virus infection (OBI) is defined as the presence of hepatitis B virus (HBV) deoxyribonucleic acid (DNA) in the liver of individuals with undetectable hepatitis B virus surface antigen (HBsAg) in the serum. The actual prevalence of OBI and its clinical relevance are not yet fully understood. OBJECTIVE: To evaluate the prevalence of HBV DNA in liver biopsies of HBsAg-negative patients with chronic liver disease of different etiologies in a referral center in Brazil and compare two different HBV DNA amplification protocols to detect HBV. DESIGN AND SETTING: This cross-sectional observational study was conducted at the Liver Outpatient Clinic, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil, between January 2016 and December 2019. METHODS: HBV DNA was investigated in 104 liver biopsy samples from individuals with chronic liver disease of different etiologies, in whom HBsAg was undetectable in serum by nested-polymerase chain reaction (nested-PCR), using two different protocols. RESULTS: OBI, diagnosed by detecting HBV DNA using both protocols, was detected in 6.7% of the 104 individuals investigated. Both protocols showed a good reliability. CONCLUSION: In addition to the differences in the prevalence of HBV infection in different regions, variations in the polymerase chain reaction technique used for HBV DNA amplification may be responsible for the large variations in the prevalence of OBI identified in different studies. There is a need for better standardization of the diagnostic methods used to diagnose this entity. BACKGROUND (OBI (HBV (DNA (HBsAg understood OBJECTIVE HBsAgnegative negative SETTING crosssectional cross sectional Clinic Clínicas Gerais Horizonte MG 201 2019 METHODS 10 nestedpolymerase nested nestedPCR, nestedPCR PCR , (nested-PCR) RESULTS 67 6 7 6.7 reliability CONCLUSION regions studies entity 20 1 (nested-PCR 6. 2

RESUMO Introdução O carcinoma hepatocelular (CHC) é a neoplasia maligna primária mais comum no fígado. O CHC se desenvolve gradualmente a partir de múltiplos estágios que controlam a proliferação e a apoptose. Na hepatocarcinogênese, múltiplas vias de sinalização já foram descritas, como a via Hedgehog (Hh). No entanto, poucos estudos investigaram a expressão de proteínas Hh como um potencial fator prognóstico no CHC humano. Este estudo teve como objetivo investigar a expressão da proteína Shh no CHC e correlacionar com características prognósticas clínicas e morfológicas do tumor. Métodos A expressão imuno-histoquímica da proteína Shh em tumor e parênquima cirrótico foi realizada em 36 amostras de CHC de pacientes submetidos a transplante hepático no Hospital das Clínicas - UFMG. Correlação entre a expressão e etiologia do tumor Shh, número de nódulos, tamanho dos nódulos, níveis de proteína alfa-feto (AFP), pontuação MELD, diferenciação tumoral e invasão vascular foram realizadas. Resultados Em nosso estudo, a marcação da proteína Shh aumentou gradualmente do parênquima normal para o cirrótico e neoplásico. Grau de diferenciação tumoral e invasão vascular foram correlacionados com alta expressão da proteína Shh (p = 0,014 ep = 0,003, respectivamente). As demais variáveis não apresentaram correlação estatisticamente significativa com a marcação de Shh. Conclusão A via Hedgehog tem importância na hepatocarcinogênese. O estudo imuno-histoquímico da via de sinalização Hh pode ter um papel promissor como fator prognóstico para CHC, principalmente devido à correlação positiva entre a expressão de Shh e o grau de diferenciação tumoral e invasão vascular.

ABSTRACT Introduction Hepatocellular carcinoma (HCC) is the most common primary malignant neoplasm in the liver. HCC develops gradually from multiple stages that control proliferation and apoptosis. In hepatocarcinogenesis, multiple signaling pathways were already described, such as the Hedgehog pathway (Hh). However, few studies have investigated the expression of Hh proteins as a potential prognostic factor in human HCC. This study aimed to investigate the expression of the Shh protein in HCC and to correlate with clinical and morphological prognostic characteristics of the tumor. Methods Immunohistochemical expression of Shh protein in tumor and cirrhotic parenchyma was performed in 36 HCC samples from patients who underwent liver transplantation at Clinical Hospital - UFMG. Correlation between the Shh tumor expression and etiology, number of nodules, size of the nodules, levels of alpha-fetus-protein (AFP), MELD score, tumor differentiation, and vascular invasion were performed. Results In our study, Shh protein labeling gradually increased from the normal to the cirrhotic and neoplastic parenchyma. Degree of tumor differentiation and vascular invasion were correlated with high Shh protein expression (p = 0.014 and p = 0.003, respectively). The other variables did not show a statistically significant correlation with Shh labeling. Conclusion Hedgehog pathway has importance in hepatocarcinogenesis. The immunohistochemical study of the Hh signaling pathway may have a promising role as a prognostic factor for HCC, especially due to the positive correlation between the Shh expression and the degree of tumor differentiation and invasion vascular.

ABSTRACT BACKGROUND: Occult hepatitis B virus infection (OBI) is defined as the presence of hepatitis B virus (HBV) deoxyribonucleic acid (DNA) in the liver of individuals with undetectable hepatitis B virus surface antigen (HBsAg) in the serum. The actual prevalence of OBI and its clinical relevance are not yet fully understood. OBJECTIVE: To evaluate the prevalence of HBV DNA in liver biopsies of HBsAg-negative patients with chronic liver disease of different etiologies in a referral center in Brazil and compare two different HBV DNA amplification protocols to detect HBV. DESIGN AND SETTING: This cross-sectional observational study was conducted at the Liver Outpatient Clinic, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil, between January 2016 and December 2019. METHODS: HBV DNA was investigated in 104 liver biopsy samples from individuals with chronic liver disease of different etiologies, in whom HBsAg was undetectable in serum by nested-polymerase chain reaction (nested-PCR), using two different protocols. RESULTS: OBI, diagnosed by detecting HBV DNA using both protocols, was detected in 6.7% of the 104 individuals investigated. Both protocols showed a good reliability. CONCLUSION: In addition to the differences in the prevalence of HBV infection in different regions, variations in the polymerase chain reaction technique used for HBV DNA amplification may be responsible for the large variations in the prevalence of OBI identified in different studies. There is a need for better standardization of the diagnostic methods used to diagnose this entity.

RESUMO CONTEXTO: A hepatocarcinogênese é um processo de múltiplas etapas que leva a alterações genéticas nos hepatócitos, resultando em neoplasia. No entanto, os mecanismos da transformação maligna parecem diferir amplamente. Conhecer os mecanismos da carcinogênese é fundamental para o desenvolvimento de novos métodos de tratamento e prevenção. OBJETIVO: O objetivo deste estudo é analisar a imunoexpressão da proteína B-Raf em explantes de carcinoma hepatocelular (CHC), em tecido cirrótico relacionado à hepatite C adjacente e em fígados normais. Também analisamos a imunoexpressão com características histopatológicas prognósticas relacionadas ao CHC. MÉTODOS: Foram estudados fígados de 35 pacientes com CHC relacionado à cirrose por vírus C submetidos a transplante hepático ou hepatectomia no Hospital das Clínicas &ndash; UFMG e 25 fígados normais de arquivos de necropsia. Os tumores foram classificados de acordo com tamanho do tumor, invasão vascular e grau de diferenciação. A expressão de B-Raf foi determinada por imunohistoquímica. RESULTADOS: B-Raf foi fortemente expresso no citoplasma do parênquima cirrótico em 17,1% dos casos e em 62,9% das amostras de CHC. A forte expressão da proteína B-Raf foi associada ao tecido tumoral (P<0,0001; OR=8,18 (2,62&ndash;26,63)). Todos os fígados normais apresentaram expressão fraca ou negativa para B-Raf. Não houve associação significativa entre os escores B-Raf e o grau de diferenciação do tumor (P=0,9485), tamanho do tumor (P=0,4427) ou invasão vascular (P=0,26666). CONCLUSÃO: Encontramos diferença na imunoexpressão da proteína B-Raf em fígados normais, nas áreas de cirrose por HCV e no hepatocarcinoma. Não encontramos associação entre a expressão de B-Raf e marcadores histopatológicos de progressão tumoral. Nossos dados sugerem que o B-Raf pode desempenhar um papel importante na carcinogênese inicial do CHC. Estudos maiores são necessários para validar essas observações.

ABSTRACT BACKGROUND: Hepatocarcinogenesis is a multistep process that lead to genetic changes in hepatocytes resulting in neoplasia. However, the mechanisms of malignant transformation seem to differ widely. To know carcinogenesis mechanisms is essential to develop new treatment and prevention methods. OBJECTIVE: The aim of this study is to analyze B-Raf protein immunoexpression in explants with hepatocellular carcinoma (HCC) related to hepatitis C (HCV), in adjacent cirrhotic tissue and in normal livers. We also associated the immunoexpression with known HCC related histopathogical prognostic features. METHODS: Livers from 35 patients with HCV related cirrhosis and HCC that underwent liver transplantation or hepatectomy at Clinical Hospital &ndash; UFMG and 25 normal livers from necropsy archives were studied. Tumors were classified according to: tumor size, vascular invasion and differentiation grade. B-Raf protein expression was determined by immunohistochemistry. RESULTS: B-Raf was strongly expressed in the HCV cirrhotic parenchyma cytoplasm of 17.1% cases and in 62.9% of HCC samples. Strong B-Raf protein staining was associated with tumor tissue (P<0.0001; OR=8.18 (2.62&ndash;26.63)). All normal livers showed weak or negative expression for B-Raf. There was no significant association among B-Raf scores and tumor differentiation grade (P=0.9485), tumor size (P=0.4427) or with vascular invasion (P=0.2666). CONCLUSION We found B-Raf protein immunostaining difference in normal livers, in the areas of HCV cirrhosis and in the hepatocarcinoma. We did not find association between B-Raf expression and histopathological markers of tumor progression. Our data suggests that B-Raf may play an important role in initial HCC carcinogenesis. Larger studies are needed to validate these observations.

ABSTRACT Purpose: The present study aimed at testing a new formulation of mesalazine linked to chondroitin sulfate and its components alone in the treatment of actinic proctitis in rats. Methods: Forty-seven female Wistar rats were submitted to pelvic radiation and divided into eight groups: control A, mesalazine A, chondroitin A, and conjugate A, gavage of the according substance two weeks after irradiation and sacrifice three weeks after oral treatment; control C, mesalazine C, chondroitin C, and conjugate C, sacrifice six weeks after oral treatment. The rectum was submitted to histological characterization for each of the findings: inflammatory infiltrate, epithelial degeneration, mucosal necrosis, and fibrosis. Results: The inflammatory infiltrate was more intense in chondroitin A, mesalazine A, and conjugate C. The collagen deposition was less intense in chondroitin A, and mesalazine A, and more intense in control C. Conclusions: Mesalazine and chondroitin alone were efficacious in inducing a delayed inflammatory response, hence reducing the late fibrosis. The conjugate was able to induce an ever more delayed inflammatory response.

Abstract During the yellow fever (YF) outbreak in Brazil, many cases of fulminant hepatitis were seen, although mild to moderate hepatitis was mostly observed with complete recovery. This report presents a case of late-onset hepatitis due to YF relapse. The patient sought medical attention after jaundice recurrence 40 days after the first YF hepatitis episode. This case highlights the importance of patient follow-up after the complete resolution of YF symptoms and discharge.

OBJECTIVES: This study aimed to analyze clinical and laboratory parameters and their association with long-term outcomes in patients who underwent liver transplantation for hepatocellular carcinoma treatment, according to the etiology of the underlying chronic liver disease, in order to identify predictors of response to this therapeutic modality. METHODS: Demographic, clinical, and laboratory data from a cohort of 134 patients who underwent orthotopic liver transplantation for hepatocellular carcinoma treatment at a referral center in Brazil were retrospectively selected and compared according to the etiologic group of the underlying chronic liver disease. Events, defined as tumor recurrence or death from any cause, and event-free survival were also analyzed, and multivariate analysis was performed. RESULTS: The etiologies comprised hepatitis C and B virus infection, alcohol abuse, and cryptogenic disorder. Although liver transplantation was performed outside the Milan criteria in 33.3% of the subjects, according to pathologic examination of the explanted liver, the Model for End-Stage Liver Disease score was low (<22) in most patients (70.6%) and recurrence was identified in only 10 (7.9%) patients. Events occurred in 37 patients (28.5%), and the median event-free survival was 75 months (range, 24-116 months). No difference among etiologic groups was found in the parameters analyzed, which were not independently associated with outcome. CONCLUSION: Clinical and laboratory characteristics according to etiologic groups were not different, which might have led to comparable long-term outcomes among these patient groups and failure to identify predictors that could aid in better selection of subjects for liver transplantation in the management of this cancer.

Abstract Purpose To evaluate different concentrations of ciprofloxacin to prevent infection after open fracture contaminated with S. aureus in rats using absorbable local delivery system. Methods Fifty-two Wistar rats were assigned to six groups. After 4 weeks, all animals underwent 99mTc-ceftizoxima scintigraphy evaluation, callus formation measurement and histological analysis. ANOVA, t-Student and Kruskal Wallis were used for quantitative variables statistical analysis, whereas qui square and exact Fisher were used for qualitative variables. Results Treatment using 25% and 50% of ciprofloxacin incorporated at the fracture fixation device were effective in preventing bone infection compared to control group (p<0.05). Chitosan were not effective in preventing bone infection when used alone compared to control group (p>0.05). Histological findings demonstrated bone-healing delay with 50% of ciprofloxacin. No difference in callus formation were observed (p>0.05). Conclusion Local delivery treatment for contaminated open fracture using chitosan with ciprofloxacin is effective above 25%.

RESUMO CONTEXTO: A atresia biliar representa a principal causa de colestase tratada cirurgicamente durante o período neonatal. Se a criança não for operada, ela evolui invariavelmente para cirrose biliar secundária. OBJETIVO: Avaliar, através de análise multivariada, os fatores prognósticos associados à presença de fluxo biliar e à sobrevida com fígado nativo após a realização da portoenterostomia de Kasai. MÉTODOS: O estudo analisou 117 pacientes com atresia biliar submetidos à portoenterostomia e com material histológico adequado para avaliação. O modelo de regressão logística foi utilizado para avaliar a presença de fluxo biliar. Sobrevida foi estudada através das curvas Kaplan-Meier e ajuste do modelo de Cox. RESULTADOS: Um terço dos pacientes obteve fluxo biliar e a mediana de idade à cirurgia foi de 81 dias. Idade à cirurgia, albumina, complicação pós-operatória, BASM (do inglês, biliary atresia structural malformation), arquitetura hepática, diâmetro do maior canalículo no porta hepatis e cirrose, segundo o escore de Ishak, foram as variáveis iniciais da análise multivariada. Idade à cirurgia maior que 90 dias de vida foi a única variável associada à ausência de drenagem biliar. A análise de sobrevida mostrou que as variáveis: ausência de fluxo biliar (P<0,0001), idade à cirurgia maior que 90 dias (P=0,035) e presença de BASM (P<0,0001), isoladamente, predizem morte ou necessidade de transplante hepático. Na análise multivariada, ausência de fluxo biliar (P<0,0001 HR:6,25 [IC95% 3,19; 12,22]) e presença de BASM (P=0,014 HR:2,16 [IC95% 1,17; 3,99]) mostraram-se associadas, com significância estatística, a menor sobrevida com fígado nativo. CONCLUSÃO: Idade à cirurgia maior que 90 dias foi identificada como fator de risco independente para ausência de fluxo biliar. Além disso, a presença de drenagem biliar e a ausência de malformações estruturais da atresia biliar são variáveis fundamentais para a maior sobrevida com fígado nativo.

ABSTRACT BACKGROUND: Biliary atresia represents the most common surgically treatable cause of cholestasis in newborns. If not corrected, secondary biliary cirrhosis invariably results. OBJECTIVE: To evaluate, through multivariate analysis, the prognostic factors associated with the presence of biliary flow and survival with the native liver following Kasai portoenterostomy. METHODS: The study analyzed data from 117 biliary atresia patients who underwent portoenterostomy and had suitable histological material for evaluation. A logistic regression model was used to assess the presence of biliary flow. Survival was investigated through Kaplan-Meier curves and Cox-adjusted models. RESULTS: One third of patients achieved biliary flow and the median age at surgery was 81 days. Age at surgery, albumin, postoperative complications, biliary atresia structural malformation (BASM), liver architecture, larger duct diameter at porta hepatis, and cirrhosis (Ishak score) were the initial variables for the multivariate analysis. Age at surgery >90 days was the only variable associated with the absence of biliary drainage. Survival analysis revealed that the absence of biliary flow (P<0.0001), age at surgery >90 days (P=0.035), and the presence of BASM (P<0.0001), alone, could predict death or need for liver transplantation. Multivariate analysis demonstrated that the absence of biliary flow (P<0.0001 hazard ratio [HR] 6.25, 95% confidence interval [CI] 3.19-12.22) and the presence of BASM (P=0.014 HR 2.16, 95% CI 1.17-3.99) were associated with lowest survival with the native liver. CONCLUSION: Age at surgery >90 days was associated with absence of biliary flow. The presence of biliary drainage and the absence of structural malformations are cornerstone features for higher survival rates with the native liver.

RESUMO Objetivo: Analisar as características clínicas, laboratoriais e histopatológicas e o percurso até o estabelecimento do diagnóstico e do tratamento de pacientes com carcinoma de suprarrenal (CSR). Métodos: Estudo retrospectivo com 13 pacientes tratados no serviço de oncologia pediátrica do Hospital das Clínicas da Universidade Federal de Minas Gerais (HC-UFMG) entre 2004 e 2015. Resultados: A idade ao diagnóstico variou de 1,0 a 14,8 anos (mediana: 2,0 anos). As manifestações de hipercortisolismo foram identificadas em todos os casos, e as de virilização, em todas as meninas. Todos os pacientes preencheram os critérios de Weiss para diagnóstico histopatológico de CSR. A imuno-histoquímica foi realizada em 61,5% dos casos. A maioria dos pacientes apresentou doença em estádio I (76,9%). Todos foram submetidos à ressecção tumoral total. Dois pacientes (estádios III e IV) receberam quimioterapia associada ao mitotano. O único óbito observado foi do paciente com doença em estádio IV. A probabilidade de sobrevida global para todo o grupo aos 5,0 anos foi de 92,3±7,4%. A mediana de tempo entre o início dos sintomas e o diagnóstico foi de 9,5 meses, e de 6,0 meses entre a primeira consulta e o início do tratamento. Conclusões: A baixa idade ao diagnóstico, o predomínio de casos com doença localizada e a ressecção tumoral completa - com apenas um caso de ruptura de cápsula tumoral - são possivelmente a explicação para a evolução favorável da população estudada. O longo percurso entre o início dos sintomas e o diagnóstico sugere a importância da capacitação dos pediatras para o reconhecimento precoce dos sinais e dos sintomas do CSR.

ABSTRACT Objective: To analyze clinical, laboratory and histopathological features and the path to diagnosis establishment and treatment of patients with adrenal carcinoma (AC). Methods: Retrospective study with 13 patients assisted at the pediatric oncology service of Hospital das Clínicas, Universidade Federal de Minas Gerais, Brazil, between 2004 and 2015. Results: Age at diagnosis ranged from 1.0 to 14.8 years (median: 2.0 years). Manifestations of hypercortisolism were identified in all cases and virilization in all girls. All patients met the Weiss criteria to AC histopathological diagnosis. Immunohistochemistry was performed in 61.5% of the cases. Most patients had stage I disease (76.9%). All subjects were submitted to total tumor resection. Two patients (stages III and IV disease) received chemotherapy associated to mitotane. The only death case was that of a patient with stage IV disease. The probability of overall survival for the entire group up to 5.0 years was 92.3±7.4%. The median time between the onset of symptoms and diagnosis was 9.5 months, and 6.0 months between first visit and start of treatment. Conclusions: Low age at diagnosis, predominance of cases with localized disease and complete tumor resection - with only one case of tumor capsule rupture - can possibly explain the favorable evolution of the studied population. The long period between onset of symptoms and diagnosis highlights the importance of training pediatricians for early recognition of AC signs and symptoms.

Abstract Purpose: To evaluate the effects of hyperbaric oxygenation on prevention of adhesions in the abdominal cavity after laparotomy. Methods: Fifty four rats underwent laparotomy; stitches were made in the four quadrant parietal peritoneum and abdominal cavity closure. Animals were divided into three groups: 1 - control; 2 - subjected to high pressures and oxygenation; 3 - subjected to 100% hyperbaric oxygenation. The animals in groups 2 and 3 were daily submitted to oxygenation hyperbaric chamber after surgery. On the seventh day another laparotomy, registration of procedure, assessment of adhesions and biopsies of the peritoneum were held. Professionals analyzed the videos and the biopsies. Results: Peritoneal cavity adhesions occurred in animals of three groups with no difference between them. In Group 3, the adhesions presented more fragile and vascular proliferation more pronounced, and there was no difference in comparison with the first and second groups. However, there was no significant difference in the evaluation of these parameters between the animals in groups 1 and 2. Conclusions: Postoperative hyperbaric oxygenation in rats submitted to laparotomy did not alter the frequency, but reduced the density of adhesions in the peritoneal cavity and promoted vascular proliferation. The change in atmospheric pressure alone had no influence on the results.

Resumo Objetivo O presente estudo avaliou a expressão do WWOX, sua associação com características clinicopatológicas e com a expressão do p53, ki-67 (proliferação celular) e CD31 (angiogênese) em pacientes com carcinoma invasivo de células escamosas do colo uterino, ou simplesmente câncer do colo uterino (CCE). Métodos Foram avaliadas prospectivamente pacientes com CCE no estágio IB (n = 20) e mulheres com mioma uterino, no grupo controle (n = 20). As pacientes com CCE foram submetidas à histerectomia radical e à linfadenectomia pélvica do tipo B-C1. As mulheres no grupo-controle foram submetidas à histerectomia vaginal. As amostras de tecido foramcoradas comhematoxilina e eosina para avaliação histológica e a expressão das proteínas foi detectada por imuno-histoquímico. Resultados A expressão do WWOX foi significativamente menor no tumor quando comparada com sua expressão no colo do útero benigno (p = 0,019). A expressão tumoral de CD31 foi inversamente associada à expressão de WWOX (p = 0,018). Sua expressão não foi associada à expressão tumoral de p53 e Ki-67 em pacientes com CCE (p = 0,464 e p = 0,360, respectivamente). Não houve associação entre a expressão de WWOX e o tamanho do tumor (p = 0,156), grau de diferenciação (p = 0,914), presença de invasão vascular linfática (p = 0,155), comprometimento do paramétrio (p = 0,421) ou metástase dos linfonodos pélvicos (p = 0,310) em pacientes com CCE. Conclusão Os resultados sugeriram que o WWOX pode estar envolvido na carcinogênese do CICECU e esse marcador foi associado à angiogênese tumoral.

Abstract Objective The current study evaluated the expression of WW domain-containing oxidoreductase (WWOX), its association with clinicopathological features and with p53, Ki-67 (cell proliferation) and CD31 (angiogenesis) expression in patients with invasive cervical squamous cell carcinoma (ICSCC). To the best of our knowledge, no other study has evaluated this association. Methods Women with IB stage-ICSCC (n = 20) and women with uterine leiomyoma (n = 20) were prospectively evaluated. Patients with ICSCC were submitted to type BC1 radical hysterectomy and pelvic lymphadenectomy. Patients in the control group underwent vaginal hysterectomy. Tissue samples were stained with hematoxylin and eosin for histological evaluation and protein expression was detected by immunohistochemistry studies. Results The WWOX expression was significantly lower in the tumor compared with the expression in thebenign cervix (p = 0.019). TheWWOXexpressionwas inversely associated with the CD31 expression in the tumor samples (p = 0.018). There was no association betweentheWWOXexpression with the p53 expression (p = 0.464)or the Ki-67expression (p = 0.360) in the samples of invasive carcinoma of the cervix. There was no association between the WWOX expression and tumor size (p = 0.156), grade of differentiation (p = 0.914), presence of lymphatic vascular invasion (p = 0.155), parametrium involvement (p = 0.421) or pelvic lymph node metastasis (p = 0.310) in ICSCC tissue samples. Conclusion The results suggested that WWOX may be involved in ICSCC carcinogenesis, and this marker was associated with tumor angiogenesis.

RESUMO Objetivo: avaliar morfologia e função de tecido esplênico autógeno, implantado no omento maior, 24 horas após conservação em solução de Ringer-lactato. Métodos: foram estudados 35 ratos machos, distribuídos em sete grupos (n=5): Grupo 1: sem esplenectomia; Grupo 2: esplenectomia total sem implante; Grupo 3: esplenectomia total e implante autógeno imediato; Grupo 4: esplenectomia total, preservação do baço em Ringer-lactato à temperatura ambiente, em seguida, fatiado e implantado; Grupo 5: esplenectomia total, baço fatiado e preservado em Ringer-lactato à temperatura ambiente antes de ser implantado; Grupo 6: esplenectomia total com preservação do baço em Ringer-lactato a 4°C e, em seguida, fatiado e implantado; Grupo 7: esplenectomia total e baço fatiado, para preservação em Ringer-lactato a 4°C antes de ser implantado. Após 90 dias, realizaram-se estudos cintilográficos com estanho coloidal-Tc99m (fígado, pulmão, baço ou implante e coágulo), hematológicos (eritrograma, leucometria, plaquetas), bioquímicos (eletroforese de proteínas) e anatomopatológicos. Resultados: ocorreu regeneração dos implantes esplênicos autógenos nos animais dos grupos com preservação do baço a 4ºC. A captação de estanho coloidal foi superior nos grupos 1, 3, 6 e 7 em relação aos demais. Não houve diferença nos valores hematimétricos nos sete grupos. A eletroforese de proteínas mostrou diminuição da fração gama no grupo de animais esplenectomizados em relação aos grupos operados. Conclusão: o tecido esplênico conservado em solução de Ringer-lactato à temperatura de 4ºC mantém sua estrutura morfológica e permite a recuperação funcional após ser implantado sobre o omento maior.

ABSTRACT Objective: to evaluate the morphology and function of autogenous splenic tissue implanted in the greater omentum, 24 hours after storage in Ringer-lactate solution. Methods: we divided 35 male rats into seven groups (n=5): Group 1: no splenectomy; Group 2: total splenectomy without implant; Group 3: total splenectomy and immediate autogenous implant; Group 4: total splenectomy, preservation of the spleen in Ringer-lactate at room temperature, then sliced and implanted; Group 5: total splenectomy, spleen sliced and preserved in Ringer-lactate at room temperature before implantation; Group 6: total splenectomy with preservation of the spleen in Ringer-lactate at 4°C and then sliced and implanted; Group 7: total splenectomy and the spleen sliced for preservation in Ringer-lactate at 4°C before implantation. After 90 days, we performed scintigraphic studies with Tc99m-colloidal tin (liver, lung, spleen or implant and clot), haematological exams (erythrogram, leucometry, platelets), biochemical dosages (protein electrophoresis) and anatomopathological studies. Results: regeneration of autogenous splenic implants occurred in the animals of the groups with preservation of the spleen at 4ºC. The uptake of colloidal tin was higher in groups 1, 3, 6 and 7 compared with the others. There was no difference in hematimetric values in the seven groups. Protein electrophoresis showed a decrease in the gamma fraction in the group of splenectomized animals in relation to the operated groups. Conclusion: the splenic tissue preserved in Ringer-lactate solution at 4ºC maintains its morphological structure and allows functional recovery after being implanted on the greater omentum.

Abstract Purpose: To describe a new model of actinic enteritis that does not use radiotherapy machines. Methods: Sixteen Wistar rats were divided into four groups, consisting of four animals each: control (group A), two weeks after irradiation (group B), five weeks after irradiation (group C) and eight weeks after irradiation (group D). Animals were given a 10Gy radiation from a Cobalt-60 natural source in a nuclear technology research center. Protections of the surrounding tissues were obtained through the usage of plumb devices with a hole in the center, which served as a collimator. We obtained irradiated and non-irradiated colons from each animal. Results: In group B we found an important inflammatory response in the irradiated colon, which appeared in a reduced way in group C and was minimal in group D, in which we found a relevant collagen submucosal deposition/fibrosis. In all groups, the non-irradiated colon had a lower pathological damage in comparison with the irradiated colon. Conclusion: We thus described an efficient and feasible technique for obtaining an animal model of actinic enteritis.