Captopril inhibits thioacetamide-induced chronic liver injury associated with the suppression of inflammation / hypoxia- inducible factor 1-alpha / profibrogenic axis-mediated hepatotoxicity in rats / Captopril inhibe la lesión hepática crónica inducida por tioacetamida asociada con la supresión de la inflamación/factor 1-alfa inducible por hipoxia/hepatotoxicidad mediada por el eje profibrogénico en rata

SUMMARY: Liver transplantation is the only available method to treat liver failure induced by chronic liver injury. We sought to determine whether the angiotensin-converting enzyme inhibitor, captopril, can inhibit the development of chronic liver injury induced by the hepatotoxic agent thioacetamide (TAA) in association with the suppression of inflammation (hsCRP, TNF-α, and IL-6) / hypoxia- inducible factor 1-alpha (HIF-1α) / profibrosis (TIMP-1, MMP-9, and α-SMA) axis that mediates liver injury. Therefore, the model group of rats was injected for eight weeks with 200 mg/kg TAA starting at week two. The protective group was pretreated with 150 mg/ kg captopril daily for two weeks prior to TAA injections and continued receiving both capropril and TAA agents until being humanely scrificed at week 10. We observed a substantial damage to liver tissue in the model group as demonstrated by a significant (p<0.0001) increase in blood and hepatic tissue levels of high sensitivity C-reactive protein (hsCRP), tumor necrosis factor-a (TNF-α), interleukin- 6 (L-6), HIF-1α, tissue inhibitor of metalloproteinases-1 (TIMP-1), matrix metalloproteinase-9 (MMP-9), alpha-smooth muscle actin (α-SMA), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). All these parameters were significantly (p<0.0244) protected by captopril. Also, a significant (p<0.0001) positive correlation was observed between a-SMA (profibrosis) and the serum and tissue levels of hsCRP, TNF-α, HIF-1α, TIMP-1, MMP-9, and ALT. Thus, these findings suggest that the induction of chronic liver injury by the hepatotoxic compound, TAA is associated with the upregulation of inflammation/HIF-1α/profibrosis, with captopril exhibiting beneficial hepatic pleotropic effects.

El trasplante de hígado es el único método disponible para tratar la insuficiencia hepática inducida por una lesión hepática crónica. Buscamos determinar si el inhibidor de la enzima convertidora de angiotensina, captopril, puede inhibir el desarrollo de lesión hepática crónica inducida por el agente hepatotóxico tioacetamida (TAA) en asociación con la supresión de la inflamación (hsCRP, TNF-α e IL-6) / factor inducible por hipoxia 1-alfa (HIF-1α) / profibrosis (TIMP-1, MMP-9 y α- SMA) eje que media la lesión hepática. Por lo tanto, al grupo modelo de ratas se le inyectó durante ocho semanas 200 mg/kg de TAA a partir de la semana dos. El grupo protector fue pretratado con 150 mg/kg de captopril al día durante dos semanas antes de las inyecciones de TAA y continuó recibiendo capropril y agentes TAA hasta que fue sacrificado en la semana 10. Observamos un daño sustancial en el tejido hepático en el grupo modelo, como lo demuestra un aumento significativo (p<0,0001) de los niveles en sangre y tejido hepático de proteína C reactiva de alta sensibilidad (hsCRP), factor de necrosis tumoral-α (TNF-a), interleucina-6 (L-6), HIF-1α, inhibidor tisular de metaloproteinasas-1 (TIMP-1), metaloproteinasa de matriz-9 (MMP-9), actina de músculo liso alfa (α-SMA), alanina aminotransferasa (ALT) y aspartato aminotransferasa (AST). Todos estos parámetros estaban significativamente (p<0,0244) protegidos por captopril. Además, se observó una correlación positiva significativa (p<0,0001) entre α-SMA (profibrosis) y los niveles séricos y tisulares de hsCRP, TNF-α, HIF-1α, TIMP- 1, MMP-9 y ALT. Por lo tanto, estos hallazgos sugieren que la inducción de daño hepático crónico por el compuesto hepatotóxico, TAA, está asociada con la regulación al alza de la inflamación/HIF-1α/profibrosis, con captopril exhibiendo efectos pleotrópicos hepáticos beneficiosos.

Prevalência do rearranjo do gene ROS1 em carcinoma de pulmão de células não pequenas não escamoso, EGFR e ALK negativo: análise de uma coorte brasileira / Prevalence of ROS1 gene rearrangements in EGFR-Negative and ALK-negative nonsquamous NSCLC: analysis of a Brazilian cohort

Introdução: A prevalência do câncer de pulmão tem aumentado cerca de 2% ao ano e é considerado um problema de saúde pública mundial, sendo a principal causa de morte por câncer entre homens e mulheres. O Câncer de Pulmão de Células Não Pequenas (CPCNP) representa 85-90% dos cânceres de pulmão. A detecção do rearranjo do gene ROS1, considerada um importante fator preditivo para direcionamento terapêutico, constitui uma etapa crítica no tratamento de CPCNP. Objetivo: Avaliar a prevalência do rearranjo do gene ROS1 em pacientes portadores de CPCNP não escamoso, sem mutação de EGFR ou rearranjo de ALK, diagnosticados na Região da Foz do Rio Itajaí, Estado de Santa Catarina, Brasil, no período de 02/01/2019 a 27/07/2020. Materiais e Métodos: Estudo observacional, retrospectivo e prospectivo, descrito e analítico com 95 pacientes que possuíam material de biópsia suficiente para a realização de novas análises e que não apresentavam mutação de EGFR ou rearranjo de ALK. Os pacientes com imuno-histoquímica positiva para a proteína ROS1 foram testados pelo método de FISH, utilizando-se uma sonda de DNA do tipo break-apart para o gene ROS1. Foi realizada uma análise descritiva da amostra, e os resultados foram apresentados em números absolutos e porcentagens, representados por tabelas. O teste de qui-quadrado (χ2) foi empregado para comparação das frequências entre os grupos analisados. Resultados: 52,6% foram pacientes do sexo masculino; a idade mediana foi de 64 anos; 54,7% declararam-se tabagistas; 40,0% apresentavam doença estágio IV; 29,5% apresentaram tumores com alta expressão de PD-L1. Quanto a expressão de ROS1 por imuno-histoquímica: 89,5% foram identificados como ROS1+ em 0% das células tumorais, 4,2% como ROS1+ em <70% das células, e 6,3% como ROS1+ em ≥70% das células do tumor; portanto, 10,5% apresentaram resultados positivos para expressão de ROS1. Estes pacientes foram submetidos à análise de rearranjo de ROS1 pelo método de FISH e 7 (7,4%) apresentaram resultados positivos. Conclusão: Na população estudada, a análise pelo método de FISH mostrou uma prevalência de 7,4% para rearranjos do gene ROS1

Introduction: Lung cancer prevalence has been increasing at rate of 2% per year and is considered a major public health concern worldwide, being the main cause of cancer death among women and men. Non-small cell lung cancer (NSCLC) represents 85-90% of total lung cancer. Detecting the rearrangement of the ROS1 gene is critical to the treatment of NSCLC. Objective: To assess the prevalence of the ROS1 gene rearrangement in patients diagnosed with non-squamous NSCLC patients diagnosed between January 2019 to July 2020 at Foz do Rio Itajaí, in the state of Santa Catarina, Brazil. Materials and Methods: This is a retrospective and prospective observational study Ninety-five NSCLC whose tumors were negative for EGFR mutation and ALK rearrangement and who had enough tumor tissue to carry out additional molecular analysis. Patients whose tumors were positive for ROS1 by immune-histochemistry were tested using by FISH using a break-apart DNA probe (Abbot Molecular) for the ROS1 gene. A descriptive analysis was performed and results were presented as absolute frequencies and percentages and depicted in charts. Frequencies were compared with the chi-squared test (χ2). Results: 52,6% were male, mean and median age were 65,10 and 64 years, respectively. 54,7% self-declared as smokers; 40,0% had stage IV disease; 29,5% had tumours with high expression of PD-L1. Regarding the expression of ROS1 by immunohistochemistry: 89,5% were identified as ROS1+ in 0% of cells, 4,2% as ROS1+ in <70% of cells, and 6,3% as ROS1+ in ≥70% of the cells; therefore, 10,5% displayed positive results for the expression of ROS1+. These tumors were subjected to the analysis of ROS1 rearrangement by FISH and 7 (7,4%) were positive. Conclusion: We observed 7,4% prevalence for ROS1 gene rearrangements in this pre-selected population

Análise clínico-patológica e qualitativa da expressão de mediadores inflamatórios teciduais no líquen plano oral / Clinicopathological and qualitative analysis of the expression of tissue inflammatory mediators in oral lichen planus

O líquen plano oral (LPO) é uma condição imuno-inflamatória mucocutânea crônica que ainda possui etiologia e patogênese desconhecidas. Estudos mostrando a participação de citocinas no LPO, em especial, interleucinas (IL)-6, IL-17 e IL-18, são escassos, assim como a correlação das características clínicas e histológicas das lesões de LPO com a presença destes mediadores inflamatórios. Todas as lesões de LPO e de lesões liquenoides orais (LLO) foram revisadas a partir do arquivo do laboratório de Patologia Bucal da Faculdade de Odontologia da Universidade do Estado do Rio de Janeiro e as características clínico-patológicas dos casos foram analisadas. Foram selecionados 40 casos de LPO para realização adicional de reações imuno-histoquímicas para IL-6, IL-17 e IL-18. A amostra total foi composta por 221 casos e mostrou que o LPO apresentou predileção por mulheres adultas, mais frequentemente acometidas pelo padrão reticular e com lesões localizadas predominantemente na mucosa jugal. Os 40 casos selecionados para a avaliação imuno-histoquímica incluíram pacientes com média de idade de 53 anos, sem predileção por gênero, e com lesões localizadas preferencialmente na mucosa jugal (85%), na gengiva/mucosa alveolar (47%) e na língua (42%). Quanto ao padrão clínico, 14 pacientes (35%) mostravam lesões exclusivamente reticulares e 26 (65%) mostravam lesões reticulares associadas a lesões atrófico-erosivas. Sintomas foram relatados por 53% dos pacientes e incluíram principalmente ardência e desconforto local. A análise histológica mostrou que o epitélio das lesões mostrava espessura normal, atrófica ou hiperplásica em, respectivamente, 17 (43%), 9 (22%) e 14 (35%) casos. A presença de hiperqueratose foi observada em 21 casos (53%) e exocitose de linfócitos T CD4+ e T CD8+ estava presente em, respectivamente, 17 (42%) e 30 (75%) casos. A análise imuno-histoquímica revelou que a IL-6 foi, de forma geral, a mais expressa, tanto no epitélio, quanto no conjuntivo. A expressão de IL-17 se mostrou intensa no tecido conjuntivo, em 40% dos casos. A IL-18 mostrou intensidade mais frequente leve/moderada tanto no epitélio (40%), quanto no tecido conjuntivo (45%). A presença de exocitose mostrou relação com a maior expressão das ILs e a expressão de IL-17 foi maior no epitélio mostrando hiperqueratose. Os resultados do presente estudo mostraram que as características clínicas das lesões de LPO e de LLO são distintas e podem ser utilizadas para diferenciação entre as duas entidades. Os achados histológicos e imunohistoquímicos sugerem que as ILs estudadas mostram-se mais presentes quando há exocitose linfócitos T CD4+ e T CD8+ e que sua expressão pode ter relação com as alterações epiteliais encontradas no LPO, participando da patogênese e da modulação da expressão da doença.

Oral lichen planus (OLP) is a chronic immunoinflammatory mucocutaneous condition of unknown etiology and pathogenesis. Studies focusing on the presence of cytokines in OLP, especially interleukin (IL)-6, IL-17 and IL-18, are scarce, as well as the correlation of clinical and histological characteristics with the presence of inflammatory mediators. All lesions diagnosed as OLP and oral lichenoid lesions (OLL) were reviewed from the files of the Oral Pathology laboratory, Dental School, Rio de Janeiro State University, and their clinicopathological characteristics were analyzed. Forty cases diagnosed as OLP were selected for additional immunohistochemical reactions directed against IL-6, IL-17 e IL-18. The total sample was composed by 221 cases and showed that OLP presented a predilection for adult females, mostly affected by lesions with the reticular pattern and located in the buccal mucosa. The 40 cases selected for the immunohistochemical reactions included patients with a mean age of 53 years, with no gender predilection, and with lesions located mostly in the buccal mucosa (85%), gingiva/alveolar mucosa (47%) and tongue (42%). The clinical pattern showed reticular lesions in 14 patients (35%) and reticular and atrophic/erosive lesions in 26 patients (65%). Symptoms were reported by 53% of the patients and included mostly burning sensation and local discomfort. Histological analysis showed that the epithelial thickness was normal, atrophic, or hyperplastic in, respectively, 17 (43%), 9 (22%) and 14 (35%) cases. The presence of hyperkeratosis was observed in 21 cases (53%), and exocytosis of T CD4+ and T CD8+ lymphocytes was present in, respectively, 17 (42%) and 30 (75%) cases. Immunohistochemical analysis showed that, in general, IL-6 was the most expressed IL both in epithelium and connective tissue. IL-17 expression was considered intense in the connective tissue from 40% of the cases. IL-18 expression was considered mostly mild/moderate both in epithelium (40% of the cases) and connective tissue (45% of the cases). The presence of exocytosis was associated with a higher expression of the ILs and expression of IL-17 was higher in epithelium showing hyperkeratosis. The results from the present study showed that the clinical characteristics of OLP and OLL are distinct and can be useful in differentiating these two diagnostic entities. The histological and immunohistochemical features suggest that the studied ILs are more expressed when there is exocytosis of both T CD4+ and T CD8+ lymphocytes. Expression of the ILs can be associated with the epithelial alterations encountered in OLP, participating in the pathogenesis and modulating the expression of the disease.

Clinical-pathological and immunohistochemical evaluations of cardiac lesions in cats with chronic kidney disease / Avaliações clínico-patológica e imuno-histoquímica de lesões cardíacas em gatos com doença renal crônica

Chronic kidney disease (CKD) is characterized by irreversible morphostructural lesions that can progressively evolve to chronic renal insufficiency and kidney failure. It is known that the heart and kidneys are closely related, and that communication between these organs occurs through a variety of pathways; subtle physiological changes in one of them are compensated by the other. Histopathological cardiac evaluation through routine staining presents a limitation to identify specific or discreet lesions in the cardiomyocytes. This study aimed to evaluate serum troponin levels in cats with CKD, associated with clinical and pathological findings, as well as to correlate the morphostructural cardiac lesions to determine their distribution through macroscopic and histological assessments and anti-cardiac troponin C (cTnC) immunohistochemistry (IHC). To this end, 20 cats (18 diagnosed with CKD and two controls) were selected. Anti-human cTnC IHC was conducted after necropsy and separation in eight regions of each collected heart. Heart fragments from two cats without CKD were used as controls. The anti-human cTnC antibody is useful in detecting cardiac lesions and has shown decreased expression in cardiomyocytes of cats with CKD. Serum troponin was above the reference values in 11/18 (61.11%) animals and decreased expression for the cTnC antibody was observed in individual cardiomyocytes in 9/18 (50%) animals. It was verified that the number of regions with decreased expression for the cTnC antibody in cardiomyocytes is significantly correlated with serum troponin. The anti-human cTnC antibody has been found effective in detecting cardiac lesions and has shown decreased expression in the cardiomyocytes of cats with CKD. Correlation was observed between increased serum cTnI and loss of immunoreactivity at anti-cTnC antibody IHC in cats with CKD, which proves damage to cardiomyocytes secondary to kidney disease.(AU)

A doença renal crônica (DRC) é caracterizada por lesões morfoestruturais irreversíveis, que podem evoluir progressivamente para insuficiência renal crônica e falência renal. Sabe-se que o coração e os rins mantêm estreita relação e a comunicação entre esses órgãos ocorre por uma variedade de vias; alterações fisiológicas sutis em um desses órgãos são compensadas pelo outro. A avaliação histopatológica cardíaca mediante a colorações rotineiras são limitadas para identificar lesões específicas ou discretas em cardiomiócitos. O presente trabalho teve como objetivos avaliar os níveis séricos de troponina em gatos com DRC, associados aos achados clínico-patológicos, bem como correlacionar as lesões cardíacas morfoestruturais, a fim de determinar a distribuição destas, por meio da avaliação macroscópica, histológica e imuno-histoquímica com anti-cTnC. Neste estudo foram selecionados 20 gatos (18 diagnosticados com DRC e 2 animais controle). Para a aplicação da técnica de imuno-histoquímica anti-troponina C humana, necropsias foram realizadas e cada coração coletado separadamente em 8 regiões. Fragmentos do coração de 2 gatos sem lesão cardíaca foram utilizados como controle. O anticorpo anti-TnC humano é útil na detecção de lesões cardíacas e apresentou expressão diminuída em cardiomiócitos de gatos com DRC. Em 11/18 animais (61,11%) a troponina sérica encontrava-se acima dos valores de referência e foram observadas diminuição da expressão para anticorpo-cTnC em cardiomiócitos individuais em 9/18 (50%). Notou-se que o número de regiões com diminuição da expressão para anticorpo-cTnC em cardiomiócitos está significativamente correlacionado com a troponina sérica. O anticorpo anti-TnC humano se mostrou eficaz para detectar lesões cardíacas e demonstrou diminuição da expressão nos cardiomiócitos de gatos com DRC. Houve correlação entre o aumento da CTnI sérica e perda da imunorretividade na avaliação imuno-histoquímica com anticorpo anti-TnC em gatos com DRC o que comprova danos em cardiomiócitos secundários a doença renal.(AU)

Sociodemographic and clinical-pathological study of molecular subtitles of breast carcinoma in a reference unit of Maranhão / Estudo sociodemográfico e clínico-patológico de legendas moleculares de carcinoma de mama em uma unidade de referência do Maranhão

Abstract Objective To evaluate the distribution of the main sociodemographic and clinicalpathological characteristics in women with breast cancer according to the molecular profile by immunohistochemistry. Methods A cross-sectional, retrospective, analytical and quantitative study was performed, with an analysis of 137 medical records from January 2015 to December 2018 of women attending the High Complexity in Oncology Unit of the city of Imperatriz, state of Maranhão, Brazil. The immunohistochemical profile of tumors based on the estrogen and progesterone receptor, Human Epidermal growth factor Receptor-type 2 (HER2) overexpression and Ki67 cell proliferation indexwas defined, fromwhich six molecular subtypes were determined: luminal A, luminal B-HER2 negative, luminal B-HER2 positive, triple negative, overexpression of HER2 and inconclusive. Results A total of 52.6% of the patients were postmenopausal, mean age 52.1 years old, brown (56.2%), had a schooling level < 9 years (40%), staging > IIB (52.6%) and 23.4% hadmetastasis. Invasive ductal carcinoma accounted for 84.7%, tumor size was 2 to 5 cm (48.9%), with lymph node involvement (56.2%), axillary lymphadenectomy in 67.2%, andmastectomy in 73.7% of the patients. Themost frequentmolecular subtype was the luminal B-HER2 negative (36.5%), and the luminal A subtype showed characteristics of better prognosis when compared with the others. Conclusion It was concluded that in the association of molecular subtypes with sociodemographic and clinical-pathological characteristics, there were no statistically significant results obtained, except for complementary therapy, referring to hormone therapy, and there was a high index of metastasis at diagnosis, which was a worrying factor and indicative of failures in the screening and early diagnosis of this population.

Resumo Objetivo Avaliar a distribuição das principais características sociodemográficas e clínico-patológicas em mulheres com câncer de mama segundo o perfil molecular pela imunohistoquímica. Métodos Estudo transversal, retrospectivo, analítico, descritivo e quantitativo, com análise de 137 prontuários do período de janeiro de 2015 a dezembro de 2018 de mulheres atendidas na Unidade de Assistência da Alta Complexidade em Oncologia da cidade de Imperatriz, MA, Brasil. Foi definido o perfil imunohistoquímico dos tumores baseado na avaliação dos receptores de estrogênio e progesterona, superexpressão de HER2 e índice de proliferação celular Ki67, de onde foram determinados seis subtipos moleculares: luminal A, luminal B-HER2 negativo, luminal B-HER2 positivo, triplo negativo, superexpressão de HER2 e inconclusivo. Resultados Foi demonstrado que 52,6% das pacientes eram pós-menopausadas, com idademédia de 52,1 anos, pardas (56,2%), tinhamgrau de escolaridade < 9 anos (40%), estadiamento > IIB (52,6%) e 23,4% tinham metástase. Carcinoma ductal invasivo representou 84,7%, o tamanho tumoral foi de 2 a 5 cm (48,9%), com comprometimento linfonodal (56,2%), com linfadenectomia axilar em 67,2% e mastectomia em 73,7% das pacientes. O subtipo molecularmais frequente foi o luminal B-HER2 negativo (36,5%), e o subtipo luminal A apresentou características de melhor prognóstico em relação aos demais. Conclusão Concluiu-se que na associação dos subtipos moleculares com as características sociodemográficas e clínico-patológicas não se obteve resultados com significância estatística, exceto para terapia complementar, referente à hormonioterapia, e houve elevado índice de metástase ao diagnóstico, o que representou um fator preocupante e indicativo de falhas no rastreio e diagnóstico precoce dessa população.

Células madre adiposas humanas disminuyen el daño de la fibrosis hepática con baja persistencia de células trasplantadas en ratas / Human adipose stem cells decrease liver fibrosis damage with low persistence of transplanted cells in rats

RESUMEN: En la enfermedad hepática crónica el trasplante ortotópico es la única alternativa terapéutica actual pero es limitada por falta de donantes. Ensayos con células madre adultas en daño hepático agudo evidencian promisorios resultados. El objetivo de este trabajo fue evaluar en ratas con daño hepático crónico la efectividad de la infusión de células madre adiposas humanas (CMAd-h). Ratas con fibrosis hepática inducida por tioacetamida fueron agrupadas en: grupo I control que no recibió tioacetamida ni células madre, grupo II recibió tioacetamida y suero fisiológico i.v., grupo III recibió tioacetamida y células madre adiposas 1 x 106/kg i.v. vía vena de la cola. La regeneración hepática histológica se evaluó por el index METAVIR, mientras las Macrophagocytus stellatus, células estrelladas a- SMA+ y células colágeno I+ por inmunohistoquímica; el daño funcional se evaluó por los niveles sanguíneos de los analitos Aspartato Aminotransferasa (AST), Alanina Aminotransferasa (ALT), Fosfatasa Alcalina (ALP), úrea y nitrógeno ureico (BUN) y hemograma. Los resultados muestran atenuación del daño estructural hepático evidenciado por disminución de los nódulos, del grado de lesión histológica en el score Metavir, y disminución de Macrophagocytus stellatus, células a-SMA+ y células colágeno tipo I+; funcionalmente hay reducción moderada de AST, ALT, urea, BUN y disminución moderada de células blancas pero efecto favorable sobre el volumen corpuscular media y la hemoglobina corpuscular media. Ocho semanas después de la infusión hay escasa población de CMAd-h en el hígado. En conclusión la infusión intravenosa de CMAd-h en ratas disminuye el daño funcional y estructural de la fibrosis hepática con escasa persistencia de CMAd-h en el parénquima hepático. A nuestro conocimiento este es el primer trabajo que evalúa el efecto de las CMAd-h en el modelo daño hepático crónico murino y la persistencia de las células trasplantadas.

SUMMARY: In chronic liver disease, orthotopic transplantation is the only current therapeutic alternative but it is limited due to lack of donors. Trials with adult stem cells in acute liver damage show promising results. The aim of this work was to evaluate the effectiveness of human adipose stem cell (h-ASC) infusion in rats with chronic liver damage. Rats with thioacetamide- induced liver fibrosis were grouped into: group I control that did not receive thioacetamide and h-ASC, group II received thioacetamide and saline i.v., group III received thioacetamide and h-ASC 1 x 106/ kg i.v. via tail vein. Histological liver regeneration was evaluated by METAVIR index, while Macrophagocytus stellatus (Kupffer cells), stellate cells a-SMA+ and collagen I+ cells by immunohistochemistry; functional damage was evaluated by blood levels of the analytes Aspartate Aminotransferase (AST), Alanine Aminotransferase (ALT), Alkaline Phosphatase (ALP), Urea and Blood Urea Nitrogen (BUN) and hemogram. The results show attenuation of structural liver damage evidenced by decreased nodules, degree of histologic injury on Metavir score, and decreased Macrophagocytus stellatus, a-SMA+ cells and type I+ collagen cells; functionally there is moderate reduction of AST, ALT, urea, BUN and moderate decrease of white cells but favorable effect on mean corpuscular volume and mean corpuscular hemoglobin. Eight weeks after infusion there is a small population of h-ASC in the liver. In conclusion, intravenous infusion of h-ASC in rats reduces functional and structural damage of hepatic fibrosis with low persistence of h- ASC in the liver parenchyma. To our knowledge this is the first work that evaluates the effect of h-SC in the model of chronic murine liver damage and the persistence of transplanted cells.

DEC1: a potential biomarker of malignant transformation in oral leukoplakia

Abstract The purpose of this study was to analyze the differential expression of DEC1 in oral normal mucosa (NM), oral leukoplakia (OLK) and oral squamous cell carcinoma (OSCC). Surgically excised specimens from patients with OLK (n = 47), OSCC (n = 30) and oral normal mucosa (n=11) were immunostained for DEC1. The expression of DEC1 protein was evaluated, and its association with the clinicopathological features was analyzed. The expression of DEC1 in NM, OLK and OSCC tissues increased in turn, and significant differences were observed among the groups (P < 0.0001). In terms of the association between DEC1 expression and epithelial dysplasia, DEC1 expression was lower in hyperkeratosis without dysplasia (H-OLK) than in OLK with moderate to severe dysplasia (S-OLK), and these differences were significant (p < 0.05). The expression of DEC1 in OSCC with OLK was significantly higher than that in OSCC without OLK (p < 0.01). Therefore, DEC1 could be a potential biomarker of malignant transformation in the carcinogenesis of OSCC, which may provide a new research direction for the transformation of oral potentially malignant disorders (OPMDs) into OSCC.

BMI-1 expression increases in oral leukoplakias and correlates with cell proliferation

Abstract Oral leukoplakia (OL) is a white lesion of an indeterminate risk not related to any excluded (other) known diseases or disorders that carry no increased risk for cancer. Many biological markers have been used in an attempt to predict malignant transformation; however, no reliable markers have been established so far. Objective To evaluate cell proliferation and immortalization in OL, comparing non-dysplastic (Non-dys OL) and dysplastic OL (Dys OL). Methodology This is a cross-sectional observational study. Paraffin-embedded tissue blocks of 28 specimens of Non-dys OL, 33 of Dys OL, 9 of normal oral mucosa (NOM), 17 of inflammatory hyperplasia (IH), and 19 of oral squamous cell carcinomas (OSCC) were stained for Ki-67 and BMI-1 using immunohistochemistry. Results A gradual increase in BMI-1 and K-i67 expression was found in oral carcinogenesis. The immunolabeling for those markers was higher in OSCC when compared with the other groups (Kruskal-Wallis, p<0.05). Ki-67 expression percentage was higher in OL and in IH when compared with NOM (Kruskal-Wallis/Dunn, p<0.05). Increased expression of BMI-1 was also observed in OL when compared with NOM (Kruskal-Wallis/Dunn, p<0.05). No differences were observed in expression of both markers when non-dysplastic and dysplastic leukoplakias were compared. A significant positive correlation between Ki-67 and BMI-1 was found (Spearman correlation coefficient, R=0.26, p=0.01). High-grade epithelial dysplasia was associated with malignant transformation (Chi-squared, p=0.03). Conclusions These findings indicate that BMI-1 expression increases in early oral carcinogenesis and is possibly associated with the occurrence of dysplastic changes. Furthermore, our findings indicate that both Ki-67 and BMI-1 are directly correlated and play a role in initiation and progression of OSCC.

[Artigo Parcialmente Retratado] Prevalência da infecção por Helicobacter pylori em pacientes dispépticos e associação com fatores de riscos clínicos para o desenvolvimento de adenocarcinoma gástrico / [article partial retraction] prevalence of helicobacter pylori infection in dyspeptic patients and its association with clinical risk factors for developing gastric adenocarcinoma

ABSTRACT BACKGROUND: In Brazil, particularly in the underdeveloped localities, the prevalence of Helicobacter pylori (H. pylori) infections can range up to 90%. These rates are higher in older individuals and vary by country region. H. pylori infections are linked to the development of gastric pathologies, namely mild to moderate gastritis, gastroenteritis, peptic ulcer, intestinal metaplasia, and gastric cancer. In 1994, this organism was classified by the International Agency for Research on Cancer (IARC) as pertaining to the Group 1 carcinogen for gastric adenocarcinoma etiology. Gastric cancer represents a significant public health problem, being the fourth most common malignant tumor and the second largest cause of cancer-related deaths. OBJECTIVE: To investigate the prevalence of H. pylori infection in dyspeptic patients and determine the link between clinical risk factors and gastric adenocarcinoma diagnosis. METHODS: Polymerase chain reaction (PCR) analysis was employed for molecular diagnosis of gastric tissue biopsies collected from 113 dyspeptic patients at the University Hospital of Federal University of Goiás. Molecular analyses allowed the identification of H. pylori infections. Furthermore, histopathological examinations were performed to determine the clinical risks of developing gastric malignancies. RESULTS: The test results identified 69 individuals older than 44 years, from 75 (66.4%) positive H. pylori infection samples. The prevalence of gastric adenocarcinoma in this study was 1.3%. Among the infected patients, six (8.2%) had high risk, and 67 (91.8%) had a low risk of developing gastric cancer (P<0.05). CONCLUSION: This study shows a high prevalence of H. pylori infection and identifies its contribution to gastric inflammations, which in the long term are manifested in high-risk clinical factors for the development of gastric adenocarcinoma.

RESUMO CONTEXTO: No Brasil, particularmente nas áreas mais pobres, a prevalência da infecção por Helicobacter pylori pode variar até 90%. Esses índices aumentam com o envelhecimento da população e são distintos entre as diferentes regiões do país. Podendo manifestar diferentes sintomatologias, essa infecção está diretamente relacionada com o desenvolvimento de patologias gástricas como gastrite leve a moderada, gastroenterites, úlcera péptica, metaplasia intestinal e principalmente, o câncer gástrico. Em 1994 a bactéria foi categorizada pela International Agency for Research on Cancer (IARC) como carcinógeno do Grupo 1 para adenocarcinoma gástrico, tipo de câncer que representa um importante problema de saúde pública, sendo o quarto tumor maligno mais comum e a segunda maior causa de mortes por câncer no mundo. OBJETIVO: Analisar a prevalência da bactéria em pacientes dispépticos e avaliar a associação de fatores de risco clínicos para desenvolvimento de adenocarcinoma gástrico. MÉTODOS: Biópsias de tecido gástrico coletadas de 113 pacientes dispépticos, atendidos no Hospital das Clínicas da Universidade Federal de Goiás, foram submetidas a diagnóstico molecular por meio de Reação em Cadeia da Polimerase, para identificação da infecção por Helicobacter pylori, e exame histopatológico, para avaliar o risco clínico de desenvolvimento de adenocarcinoma gástrico. RESULTADOS: Foram diagnosticadas 75 (66,4%) amostras positivas para infecção por Helicobacter pylori, sendo 69 indivíduos maiores de 44 anos de idade. A prevalência do adenocarcinoma gástrico nesse estudo foi de 1,3% e dentre os pacientes positivos para a infecção bacteriana seis (8,2%) possuem alto risco e 67 (91,8%) baixo risco de desenvolver esse tipo de câncer (P<0,05). CONCLUSÃO: Esse estudo mostra uma alta prevalência da infecção por H. pylori na população estudada e identifica sua intrínseca contribuição para inflamações gástricas, que a longo prazo se manifestam em fatores clínicos de alto risco para o desenvolvimento de adenocarcinoma gástrico.

Análise imunoistoquímica da expressão de PD-L1 em células tumorais e células T nos carcinomas de células escamosas do pênis / Immunohistochemical analysis of PD-L1 expression in tumor cells and T cells in penile squamous cell carcinomas

Introdução: A neoplasia do pênis é uma doença rara, sendo mais frequente nos países em desenvolvimento. Representa 0,4%dos tumores malignos dos homens nos EUA e 2,1% no Brasil, sendo mais prevalente nas regiões norte e nordeste. O carcinoma epidermóide representa aproximadamente 95% das neoplasias do pênis e sua ocorrência está ligada à presença de fimose, higiene inadequada e infecções virais. O PD-1 e seus ligante PD-L1 são proteínas de membrana com papel crucial na regulação da resposta imune. O aumento da expressão de PD-L1 parece ser um mecanismo de evasão do sistema imune do hospedeiro pelas células tumorais e tem sido alvo de diversos estudos. Objetivos: estabelecer a frequência da Expressão da proteína PD-L1 e das células T nos carcinomas de pênis, correlacionando os aspectos demográficos e clínico-patológicos com a expressão doe as tendências do câncer de pênis no Estado do Ceará. Métodos: Tratou-se de estudo retrospectivo, transversal, clínico-patológico e imunoistoquímico de câncer de pênis em pacientes tratados e em acompanhamento no Hospital do Câncer Haroldo Juaçaba do Instituto do Câncer do Ceará (HHJ/ICC). A amostra constituiu-se de 84 pacientes com diagnóstico de câncer de pênis acompanhados no HHJ/ICC, entre 2002 e 2014. Os dados foram obtidos através de revisão dos prontuários e do preenchimento de formulário elaborado pelos pesquisadores. Foi preparado TMA a partir de amostras dos blocos de parafina dos tumores. A imunoistoquímica foi realizada a partir das lâminas do TMA. Os dados foram tabulados, armazenados e processados através do programa estatístico Statistical Package for the Social Science (SPSS) Versão 22.0. Resultados: A média de idade foi de 67,6 anos com a maioria oriunda do interior do Estado (65,5%) e casados (68,7%). Amputação parcial foi a cirurgia mais realizada (82,14%) e a maioria dos tumores tinha até 5cm (77,4%). Tivemos cerca de 31% pT1 e 21,5% pN positivo. O subtipo usual foi o prevalente (67,9%) e os carcinomas moderadamente diferenciados (61,9%) tiveram maior representatividade. Houve recidiva em cerca de 21% dos casos. Quanto ao PD-L1 houve expressão em 22,6% dos casos. Houve significância estatística para a associação entre a expressão de PD-L1 com Células Inflamatórias (p=0,000), CD4 (p=0,001), CD8 (p=0,012) e estádio pT (p=0,02). Também houve para a associação entre a expressão de Células Inflamatórias com CD4 (p=0,004), CD8 (p=0,000), estádio pT (p=0,001), grau histológico (p=0,03) e diferenciação histológica (p=0,04). Quanto ao prognostico, a recidiva (p=0,017), o estádio pT (p=0,027) e o grau histológico (p=0,020) mostraram-se como fatores de impacto na sobrevida. Conclusão: Trata-se de uma neoplasia que acomete mais a sexta década da vida. A recidiva, o estadiamento patológico (pT) e o grau histológico ratificaram-se como fatores prognósticos relevantes na sobrevida. A expressão de PD-L1 em tumores avançados reforça a importância de continuar a pesquisa para investigação adicional de anti-PD-1 e imunoterapêuticos anti-PD-L1 em pacientes com neoplasia peniana avançada. Reforça-se ainda que por se tratar de um problema de saúde pública, campanhas de conscientização quanto aos sintomas devem ser incentivadas para que se possa fazer o diagnóstico mais precocemente, o que poderia diminuir as taxas de incidência de tumores em estágio avançado e propiciar cirurgias menos mutilantes

Introduction: Penile neoplasia is a rare disease, being more frequent in developing countries. It represents 0.4% of the malignant tumors of the men in the USA and 2.1% in Brazil, being more prevalent in the north and northeast regions. Epidermoid carcinoma accounts for approximately 95% of penile neoplasms, and its occurrence is linked to the presence of phimosis, inadequate hygiene and viral infections. PD-1 and its PD-L1 linkers are membrane proteins with crucial role in the regulation of the immune response. Increased PD-L1 expression appears to be a mechanism for evading the host immune system by tumor cells and has been the subject of several studies. Objectives: To establish the frequency of PD-L1 protein and T-cell expression in penile carcinomas, correlating demographic and clinical-pathological aspects with the expression and trends of penile cancer in the State of Ceara. Methods: Retrospective, cross-sectional, clinical-pathological and immunohistochemical study of penile cancer in patients treated and followed up at Haroldo Juaçaba Cancer Hospital of the Cancer Institute of Ceara (HHJ / ICC). The sample consisted of 84 patients with a diagnosis of penile cancer in the HHJ / ICC, between 2002 and 2014. The data were obtained by reviewing the medical records and filling out a form prepared by the researchers. TMA was prepared from samples of the paraffin blocks of the tumors. Immunohistochemistry was performed from the TMA slides. Data were tabulated, stored and processed using the statistical program Statistical Package for Social Science (SPSS) Version 22.0. Results: The mean age was 67.6 years, with the majority coming from the interior of the State (65.5%) and married (68.7%). Partial amputation was the most performed surgery (82.14%) and the majority of tumors had up to 5 cm (77.4%). We had about 31% pT1 and 21.5% pN positive. The usual subtype was the prevalent one (67.9%) and the moderately differentiated carcinomas (61.9%) were more representative. There was recurrence in about 21% of the cases. Regarding PD-L1, there was 22.6% of cases. There was statistical significance for the association between PD-L1 expression with Inflammatory Cells (p = 0.000), CD4 (p = 0.001), CD8 (p = 0.012) and pT stage (p = 0.02). There was also an association between the expression of CD4 (p = 0.004), CD8 (p = 0.000), pT (p = 0.001), histological grade (p = 0.03) and histological differentiation, 04). As for the prognosis, the relapse (p = 0.017), the pT stage (p = 0.027) and the histological grade (p = 0.020) were shown as survival impact factors. Conclusion: This is a neoplasm that affects the sixth decade of life. Recurrence, pathological staging (pT) and histological grade were ratified as relevant prognostic factors in survival. The expression of PD-L1 in advanced tumors reinforces the importance of continuing the research for further investigation of anti-PD-1 and anti-PD-L1 immunotherapeutics in patients with advanced penile neoplasia. It is also reinforced that because it is a public health problem, awareness campaigns about the symptoms should be encouraged so that the diagnosis can be made earlier, which could reduce the incidence rates of advanced tumors and provide surgeries less mutilating

Imunoexpressão das tetraspaninas (CD9 E UPIII) no carcinoma urotelial de bexiga / Immunoexpression of tetraspanin (CD9 e UPIII) in bladder urothelial carcinoma

Introdução: O carcinoma urotelial da bexiga é uma doença de alta incidência e letalidade. Recentemente, tem sido caracterizado em subtipos moleculares, com diferenças prognósticas e de resposta terapêutica. As tetraspaninas estão implicadas na adesão celular e sinalização intercelular, interferindo diretamente em fenômenos como a morfogênese dos tecidos e migração. Uma vez que os subtipos moleculares luminal e basal correspondem, respectivamente, a fenótipos celulares urotelial bem diferenciado e de reserva basal, é possível que a expressão das tetraspaninas acompanhe esse racional, o que traria possibilidade de integração no cenário clínico. Metodologia: Amostras de 88 pacientes operados (cistectomia radical) por carcinoma urotelial, preservadas em parafina, foram organizadas em TMA ou cortes inteiros, submetidos a imuno-histoquímica. As reações foram analisadas por microscopia ótica convencional e acessadas quanto à intensidade de expressão - negativo, fraco e forte (escore subjetivo) e extensão de expressão ­ negativo, focal e difuso (baseado na porcentagem das células positivas, em cortes de 1-50%; >50%). Os tumores foram classificados em Luminal, Basal e Sem tipo definido, a partir da expressão imuno-histoquímica de GATA3, CK5/6 e CK14. A expressão citoplasmática das tetraspaninas (CD9 e UPIII) foi avaliada em tecidos tumoral e urotelial não neoplásico pareado, e correlacionadas com parâmetros patológicos e clínicos. Resultados: A idade média foi de 66.9 anos, com relação H:M de 2:1 e 72% de tabagistas ou ex-tabagistas. A maior parte (62%) das cirúrgicas foram indicadas por doença músculo-invasiva já ao diagnóstico e o restante por doença não músculo-invasiva extensa/recorrente ou progredida para carcinoma músculo invasor, sendo o estadiamento final na cistectomia 14%

Introduction: Urothelial carcinoma of the bladder is a disease with high incidences and letality. It has recently been carachterized in molecular subtypes, with differences regarding prognosis and response to treatment. Tetraspanins are implicated in cell adhesion and intercellular signaling, directly interfering in phenomena such as tissue morphogenesis and migration. Considering that luminal and basal molecular subtypes correspond, repectively, to well differentiated urotelial and basal reserve cell phenotypes, it is possible that tetraspanin expression may follow this rationale, which could bare potential to integration into the clinical scenario. Methodology: Paraffin-preserved samples from 88 patients submitted to radical cystectomy for urothelial carcinoma were organized into TMAs or whole sections, labeled with immunohistochemistry. Reactions were analyzed by conventional optical microscopy and accessed as to expression intensity ­ negative, weak or strong (subjective scoring) and extent ­ negative, focal or diffuse (based on percentage of positive cells in cut-offs of 1-50%; >50%). Tumors were classified into Luminal, Basal and Non-type based on immunoexpression of GATA3, CK14 and CK5/6. Cytoplasmic expression of tetraspanins was evaluated in neoplastic and non-neoplastic urotelial tissue and correlated with clinical and morphological parameters. Results: Mean age was 66.9 years, with M:F ratio of 2:1 and 72% smokers or ex-smokers. The majority (62%) of surgeries was indicated because of muscle-invasive disease at presentation and the rest for extensive/recurrent non muscle-invasive disease or progression to invasive carcinoma (final pathological stage at cystectomy: 14%

Chronic prostatitis alters the prostatic microenvironment and accelerates preneoplastic lesions in C57BL/6 mice

BACKGROUND: Chronic prostatitis has been supposed to be associated with preneoplastic lesions and cancer development. The objective of this study was to examine how chronic inflammation results in a prostatic microenvironment and gene mutation in C57BL/6 mice. METHODS: Immune and bacterial prostatitis mouse models were created through abdominal subcutaneous injection of rat prostate extract protein immunization (EAP group) or transurethral instillation of uropathogenic E. coli 1677 (E. coli group). Prostate histology, serum cytokine level, and genome-wide exome (GWE) sequences were examined 1, 3, and 6 months after immunization or injection. RESULT: In the EAP and E. coli groups, immune cell infiltrations were observed in the first and last months of the entire experiment. After 3 months, obvious proliferative inflammatory atrophy (PIA) and prostatic intraepithelial neoplasia (PIN) were observed accompanied with fibrosis hyperplasia in stroma. The decrease in basal cells (Cytokeratin (CK) 5+/p63+) and the accumulation of luminal epithelial cells (CK8+) in the PIA or PIN area indicated that the basal cells were damaged or transformed into different luminal cells. Hic1, Zfp148, and Mfge8 gene mutations were detected in chronic prostatitis somatic cells. CONCLUSION: Chronic prostatitis induced by prostate extract protein immunization or E. coli infection caused a reactive prostatic inflammation microenvironment and resulted in tissue damage, aberrant atrophy, hyperplasia, and somatic genome mutation.

Macrophages subpopulations in chronic periapical lesions according to clinical and morphological aspects

Abstract: The aim of this study was to evaluate macrophage M1 and M2 subpopulations in radicular cysts (RCs) and periapical granulomas (PGs) and relate them to clinical and morphological aspects. M1 macrophages were evaluated by the percentage of CD68 immunostaining associated with the inflammatory cytokine TNF-α, and M2 macrophages, by its specific CD163 antibody. The CD68+/CD163+ ratio was adopted to distinguish between the two macrophage subpopulations. Clinical, radiographic, symptomatology, treatment, and morphological parameters of lesions were collected and a significance level of p = 0.05 was adopted for statistical analysis. The results showed that the CD68+/CD163+ ratio was higher in the RCs (median = 1.22, p = 0.002), and the highest TNF-α immunostaining scores were found in RCs (p = 0.018); in PGs, the CD68+/CD163+ ratio was lower and associated with a greater CD163+ immunostaining (median = 1.02, p <0.001). The TNF-α in cyst epithelium had a score of 3 in 10 cases and predominance of M1 macrophages by CD68+/CD163+ (median = 2.23). In addition, CD68+ cells had higher percentage of immunostaining in smaller RCs (p = 0.034). Our findings suggest that increased CD68 immunostaining associated with TNF-α cytokine in RCs results in a greater differentiation of the M1 phenotype. The higher CD163 immunostaining in PGs results in greater differentiation of the M2 phenotype. Therefore, the inflammatory state promoted by M1 macrophages is related to growth and progression of RCs; on the other hand, the immunomodulatory state of M2 macrophages is related to maintenance of PGs.

Evaluation of the profile of inflammatory cytokines, through immunohistochemistry, in the skin of patients with allergic contact dermatitis to nickel in the acute and chronic phases

Abstract: Background: Allergic contact dermatitis to ion nickel (Ni+2) is an inflammatory dermatosis, common in industrialized countries. It involves the activation of nickel-specific T-cells, followed by proliferation and induction of a mixed profile of both proinflammatory and regulatory cytokines, suggesting that several T-cell subtypes (helper - Th and cytotoxic - Tc) are involved. A broader understanding of the cytokine profile may lead to new therapeutic approaches. Objectives: This study aimed to analyze the cytokines TNF-α, INF-γ, IL-2, IL-4, IL-10, IL-13, IL-17 and IL-23 using the immunohistochemistry technique in order to try to identify their prevalence in chronic and acute eczema of patients with allergic contact dermatitis to Ni+2. Methods: We performed an immunohistochemical study for eight cytokines in 20 patients with Ni+2 allergic contact dermatitis, biopsied at the site of chronic eczema, triggered by the patient's daily contact with Ni+2, and at the site of acute eczema caused by nickel sulfate, 48 hours after applying the contact test. Results: The stained samples showed positive results for the eight cytokines studied. TNF-α, IFN-γ, IL-4, IL-13 and IL-17 had a higher prevalence in chronic eczema, IL-2 and IL-23 in acute eczema, and IL-10 presented a similar prevalence in both acute and chronic eczema. However, these prevalences were statistically significant only for IL-4 and IL-13. Study Limitations: Small sample size. Conclusions: In chronic and acute eczema, we observed the presence of a mixed cytokine profile of the T cell subtypes (Th/Tc), suggesting that the responses are expressed at the same time.

A pioneering RET genetic screening study in the State of Ceará, Brazil, evaluating patients with medullary thyroid cancer and at-risk relatives: experience with 247 individuals

ABSTRACT Objective: Initial diagnosis of medullary thyroid carcinoma (MTC) is frequently associated with advanced stages and a poor prognosis. Thus, the need for earlier diagnoses and detection in relatives at risk for the disease has led to increased use of RET genetic screening. Subjects and methods: We performed RET screening in 247 subjects who were referred to the Brazilian Research Consortium for Multiple Endocrine Neoplasia (BRASMEN) Center in the State of Ceará. Direct genetic sequencing was used to analyze exons 8, 10, 11, and 13-16 in MTC index cases and specific exons in at risk relatives. Afterward, clinical follow-up was offered to all the patients with MTC and their affected relatives. Results: RET screening was performed in 60 MTC index patients and 187 at-risk family members. At the initial clinical assessment of the index patients, 54 (90%) were diagnosed with apparently sporadic disease and 6 (10%) diagnosed with hereditary disease. After RET screening, we found that 31 (52%) index patients had sporadic disease, and 29 (48%) had hereditary disease. Regarding at-risk relatives, 73/187 were mutation carriers. Mutations in RET codon 804 and the rare p.M918V mutation were the most prevalent. Conclusions: Performing RET screening in Ceará allowed us to identify a different mutation profile in this region compared with other areas. RET screening also enabled the diagnosis of a significant number of hereditary MTC patients who were initially classified as sporadic disease patients and benefited their relatives, who were unaware of the risks and the consequences of bearing a RET mutation.

Immunohistopathological and Biochemical Study of the Effects of Dead Nettle (Urtica Dioica) Extract on Preventing Liver Lesions Induced by Experimental Aflatoxicosis in Rats / Estudio inmunohistopatológico y bioquímico de los efectos del extracto de la ortiga mayor (Urtica dioica) en la prevención de lesiones hepáticas inducidas por aflatoxicosis experimental en ratas

ABSTRACT Objective: Aflatoxicosis is a mycotoxicosis infection with an acute or chronic course that forms due to aflatoxins (AFs) in humans and animals. Aflatoxins primarily affect the liver and can lead to histopathological necrosis, fibrosis and hepatocarcinogenesis of the organ. This paper studied the preventive effects of dead nettle leaf (Urtica dioica leaf; UDL) extract on liver lesions that were induced by experimental aflatoxicosis in rats. Methods: A total of 30 rats were separated into three groups of 10 rats each. Experimental group A (control) received normal rat food, experimental group B (AFB1) received 2 mg/kg of AF, and experimental group C (AFB1 + UDL extract) received 2 mg/kg of AF + 2 ml/rat/day of UDL extract. After three months of experimentation, blood and tissue samples were taken from the rats by necropsy to perform chemical and histopathological analyses. Results: According to the biochemical and histopathological findings, antioxidant system activity increased and lipid peroxidation and liver enzyme levels decreased in the group that received UDL extract. Conclusion: The extract of UDL had hepatoprotective effects against aflatoxicosis.

RESUMEN Objetivo: La aflatoxicosis es una infección por micotoxicosis con un curso agudo o crónico producido por aflatoxinas (AF) en seres humanos y animales. Las aflatoxinas afectan principalmente el hígado y pueden conducir a necrosis histopatológica, fibrosis o hepatocarcinogénesis del órgano. En este trabajo se estudiaron los efectos preventivos del extracto de la hoja de ortiga mayor (Urtica dioica l; UDL) sobre las lesiones hepáticas inducidas por aflatoxicosis experimental en ratas. Métodos: Un total de 30 ratas se separaron en tres grupos de 10 ratas cada una. EL grupo experimental A (control) recibió comida normal de ratas; el grupo experimental B (AFB1) recibió 2 mg/kg de AF; y el grupo experimental C (AFB1 + extracto de UDL) recibió 2 mg/kg de AF + 2 ml/rata/día de extracto de UDL. Después de tres meses de experimentación, se tomaron muestras de sangre y tejidos de las ratas en una necropsia encaminada a realizar análisis químicos e histopatológicos. Resultados: Según los hallazgos bioquímicos e histopatológicos, la actividad del sistema antioxidante aumentó, y la peroxidación del lípido y los niveles de la enzima del hígado disminuyeron en el grupo que recibió el extracto de UDL. Conclusión: El extracto de UDL tuvo efectos hepatoprotectores contra la aflatoxicosis.

Evaluation of apoptosis indexes in currently used oral alpha-blockers in prostate: a pilot study

ABSTRACT Objectives: Apoptosis effect of oral alpha-blockers is known in the prostate. Apoptosis index of silodosin has not been proved, yet. Aims are to present apoptosis index of silodosin in prostate and to compare this with other currently used alpha-blocker's apoptosis indexes together with their clinical effects. Materials and Methods: Benign prostatic hyperplasia (BPH) patients were enrolled among those admitted to urology outpatient clinic between June 2014 and June 2015. Study groups were created according to randomly prescribed oral alpha-blocker drugs as silodosin 8mg (Group 1; n=24), tamsulosin 0.4mg (Group 2; n=30), alfuzosin 10mg (Group 3; n=25), doxazosin 8mg (Group 4; n=22), terazosin 5mg (Group 5; n=15). Pa- tients who refused to use any alpha-blocker drug were included into Group 6 as control group (n=16). We investigated apoptosis indexes of the drugs in prostatic tissues that were taken from patient's surgery (transurethral resection of prostate) and/or prostate biopsies. Immunochemical dyeing, light microscope, and Image Processing and Analy- sis in Java were used for evaluations. Statistical significant p was p<0.05. Results: There were 132 patients with mean follow-up of 4.2±2.1 months. Pathologist researched randomly selected 10 areas in each microscope set. Group 1 showed statisti- cal significant difference apoptosis index in immunochemical TUNEL dyeing and im- age software (p<0.001). Moreover, we determined superior significant development in parameters as uroflowmetry, quality of life scores, and international prostate symptom score in Group 1. Conclusions: Silodosin has higher apoptosis effect than other alpha-blockers in prostate. Thus, clinic improvement with silodosin was proved by histologic studies. Besides, static factor of BPH may be overcome with creating apoptosis.

Expressions of TOLL-like receptor 4 (TLR-4) and matrix metalloproteinase 9 (MMP-9)/Tissue inhibitor of metalloproteinase 1 (TIMP-1) in pulmonary blood vessels with chronic obstructive pulmonary diseases and their relationships with pulmonary vascular remodelling

SUMMARY OBJECTIVE: This study aims at investigating the expressions of TOLL-like receptor 4 (TLR-4) and matrix metalloproteinase 9 (MMP-9)/ tissue inhibitor of metalloproteinase 1 (TIMP-1) in pulmonary blood vessels with chronic obstructive pulmonary disease (COPD) and their relationships with pulmonary vascular remodelling (PVR). METHODS: 60 para-tumour tissues were divided into the COPD group and the control group (n=30); the inflammations, pulmonary artery wall area/total artery area (WA%), and wall thickness/vascular outer diameter (WT%) were compared. The expressions of TLR-4, MMP-9/TIMP-1, and PCNA in pulmonary vascular smooth muscle cells were detected, and their relationships with PVR were then analysed. RESULTS: The inflammations (1.6±0.8), WA% (44.0±6.4), and WT% (27.3±3.3) in the COPD group were higher than in the control group (0.3±0.5, 26.1±2.8, 15.6±1.8), and the expressions of TLR-4 (31.4±147) and MMP-9/TIMP-1 (2.2±2.6) were increased compared to the control group (4.7±4.5, 1.9±12). Correlation analysis: TLR-4 and MMP-9/TIMP-1 were positively correlated with the inflammations (r=0.18, P<0.01), WA% (r=0.68, P<0.01), and WT% (r=0.73, P<0.01), as well as positively correlated with the expression of PCNA (r=0.44, P<0.01); the upregulation of TLR-4 was positively correlated with the expressions of MMP-9 and TIMP-1. CONCLUSIONS: The upregulation of TLR-4 in the pulmonary arterial smooth muscle cells of COPD patients could promote the inflammations and the MMP-9 expression, thus causing abnormal degradation of extracellular matrix, so it played an important role in the process of PVR.

RESUMO OBJETIVO: Este estudo tem como objetivo investigar as expressões de TOLL-like receptor 4 (TLR-4) e metaloproteinase 9 da matriz (MMP-9)/inibidor de tecido da metaloproteinase 1 (TIMP-1) em vasos sanguíneos pulmonares com doença pulmonar obstrutiva crônica (DPOC) e suas relações com o remodelamento vascular pulmonar (PVR). MÉTODOS: Sessenta tecidos paratumorais foram divididos em grupo COPD e o grupo controle (n = 30). Foram comparadas as inflamações, área da parede da artéria pulmonar/área da artéria total (WA%) e espessura da parede/diâmetro externo vascular (WT%). As expressões de TLR-4, MMP-9/TIMP-1 e PCNA em células de músculo liso vascular pulmonar foram detectadas, e suas relações com PVR foram então analisadas. RESULTADOS: As inflamações (1,6 ± 0,8), WA% (44,0 ± 6,4) e WT% (27,3 ± 3,3) no grupo COPD foram maiores que no grupo controle (0,3 ± 0,5; 26,1 ± 2,8; 15,6 ± 1,8). E as expressões de TLR-4 (31,4 ± 14,7) e MMP-9/TIMP-1 (2,2 ± 2,6) foram aumentadas em relação ao grupo controle (4,7 ± 4,5, 1,9 ± 1,2). Na análise de correlação, TLR-4 e MMP-9/TIMP-1 foram positivamente correlacionadas com as inflamações (r = 0,18; P <0,01), WA% (r = 0,68; P <0,01) e WT% (r = 0,73; P <0,01), bem como correlacionadas positivamente com a expressão de PCNA (r = 0,44; P <0,01). A elevação da TLR-4 foi correlacionada positivamente com as expressões de MMP-9 e TIMP-1. CONCLUSÕES: A regulação positiva do TLR-4 nas células do músculo liso arterial pulmonar de pacientes com DPOC poderia promover as inflamações e a expressão de MMP-9, causando assim uma degradação anormal da matriz extracelular, por isso desempenhou um papel importante no processo de PVR.