The role of TNF-α and NFkß in an experimental model of intestinal carcinogenesis with 1, 2-dimethyhydrazine

ABSTRACT Purpose: To analyze the potential of tumor necrosis factor-α (TNF-α) and factor nuclear kappa B (NF-κB) as colorectal cancer (CRC) biomarkers in an experimental model of intestinal carcinogenesis with 1,2-dimethyhydrazine (1,2-DMH). Methods: Twenty-four male Wistar rats were divided into two groups: sham and 1,2-DMH. First, 1,2-DMH (20 mg/kg/week) was administered for 15 consecutive weeks. In the 25th week, proctocolectomy was conducted. Histopathological analysis, immunohistochemistry, and gene expression of TNF-α and NF-κB were performed. Statistical analysis was performed using GraphPad Prism. The location of aberrant crypt foci (ACF) was analyzed by Kruskal-Wallis' test. For analyses with two groups with parametric data, the t-test was used; for non-parametric data, the Mann-Whitney's test was used. P < 0.05 was considered significant. Results: The number of ACF and macroscopic lesions was significantly higher (p < 0.5) in the 1,2-DMH group compared to the sham group, and most ACF were concentrated in the distal segment of the colon. There was a statistically significant increase (p < 0.5) in protein and gene expression of TNF-α and NF-κB in the 1,2-DMH group compared to the sham group. Conclusions: Our results provide supportive evidence that TNF-α and NF-κB pathways are strongly involved in CRC development in rats and might be used as early biomarkers of CRC pathogenesis in experimental studies.

Anticarcinogenic activity of methanol extract of Melastoma malabathricum leaves is attributed to the presence of phenolics compounds and the activation of endogenous antioxidant system / Actividad anticancerígena del extracto metanólico de hojas de Melastoma malabathricum atribuida a la presencia de compuestos fenólicos y a la activación del sistema antioxidante endógeno

Melastoma malabathricum (M. malabathricum) extracts have been reported to exert various pharmacological activities including antioxidants, anti-inflammatory and antiproliferative activities. The objective of the present study was to determine the anticarcinogenic activity of its methanol extract (MEMM) against the azoxymethane (AOM)-induced early colon carcinogenesis in rats. Rats were randomly assigned to five groups (n=6) namely normal control, negative control, and treatment (50, 250 or 500 mg/kg of MEMM) groups. Colon tissues were harvested for histopathological analysis and endogenous antioxidant system determination. MEMM was also subjected to HPLC analysis. Findings showed that MEMM significantly (p<0.05) reversed the AOM-induced carcinogenicity by: i) reducing the formation of aberrant crypt foci (ACF) in colon tissues, and; ii) enhancing the endogenous antioxidant activity (catalase, superoxide dismutase and glutathione peroxidase). Moreover, various phenolics has been identified in MEMM. In conclusion, MEMM exerts the in vivo anticarcinogenic activity via the activation of endogenous antioxidant system and synergistic action of phenolics.

Se ha informado que los extractos de Melastoma malabathricum (M. malabathricum) ejercen diversas actividades farmacológicas, incluidas actividades antioxidantes, antiinflamatorias y antiproliferativas. El objetivo del presente estudio fue determinar la actividad anticancerígena de su extracto de metanol (MEMM) contra la carcinogénesis de colon temprana inducida por azoximetano (AOM) en ratas. Las ratas se asignaron al azar a cinco grupos (n=6), a saber, los grupos de control normal, control negativo y tratamiento (50, 250 o 500 mg/kg de MEMM). Tejidos de colon fueron recolectados para análisis histopatológico y determinación del sistema antioxidante endógeno. MEMM también se sometió a análisis de HPLC. Los hallazgos mostraron que MEMM invirtió significativamente (p<0.05) la carcinogenicidad inducida por AOM al: i) reducir la formación de focos de criptas aberrantes (ACF) en los tejidos del colon, y; ii) potenciar la actividad antioxidante endógena (catalasa, superóxido dismutasa y glutatión peroxidasa). Además, se han identificado varios fenólicos en MEMM. En conclusión, MEMM ejerce la actividad anticancerígena in vivo mediante la activación del sistema antioxidante endógeno y la acción sinérgica de los fenólicos.

Caracterização morfológica da abertura de glândulas colônicas usando imagens de microscopia de autofluorescência / Morphological characterization of colorectal pits using autofluorescence microscopy images

ABSTRACT BACKGROUND: Colorectal cancer is one of the most prevalent pathologies. Its prognosis is linked to the early detection and treatment. Currently diagnosis is performed by histological analysis from polyp biopsies, followed by morphological classification. Kudo's pit pattern classification is frequently used for the differentiation of neoplastic colorectal lesions using hematoxylin-eosin stained samples. Few articles have reported this classification with image software processing, using exogenous markers over the samples. The processing of autofluorescence images is an alternative that could allow the characterization of the pits from the crypts of Lieberkühn, bypassing staining techniques. OBJECTIVE: Processing and analysis of widefield autofluorescence microscopy images obtained by fresh colon tissue samples from a murine model of colorectal cancer in order to quantify and characterize the pits morphology by measuring morphology parameters and shape descriptors. METHODS: Adult male BALB/cCmedc strain mice (n=27), ranging from 20 to 30 g, were randomly assigned to four and five groups of treated and control animals. Colon samples were collected at day zero and at fourth, eighth, sixteenth and twentieth weeks after treatmentwith azoxymethane. Two-dimensional (2D) segmentation, quantification and morphological characterization of pits by image processing applied using macro programming from FIJI. RESULTS: Type I is the pit morphology prevailing between 53 and 81% in control group weeks. III-L and III-S types were detected in reduced percentages. Between the 33 and 56% of type I was stated as the prevailing morphology for the 4th, 8th and 20th weeks of treated groups, followed by III-L type. For the 16th week, the 39% of the pits was characterized as III-L type, followed by type I. Further, pattern types as IV, III-S and II were also found mainly in that order for almost all of the treated weeks. CONCLUSION: These preliminaries outcomes could be considered an advance in two-dimensional pit characterization as the whole image processing, comparing to the conventional procedure, takes a few seconds to quantify and characterize non-pathological colon pits as well as to estimate early pathological stages of colorectal cancer.

RESUMO CONTEXTO: O câncer colorretal é uma das patologias mais prevalentes. Seu prognóstico é ligado à detenção e ao tratamento precoces. Atualmente o diagnóstico é realizado por análise histológica de biópsias de pólipo, seguida de classificação morfológica. A classificação de padrões de Kudo é frequentemente utilizada para a diferenciação de lesões colorretais neoplásicas usando amostras coradas por hematoxilina-eosina. Poucos artigos relatam esta classificação com utilização de processamento por software de imagem, utilizando marcadores exógenos sobre as amostras. O processamento de imagens de autofluorescência é uma alternativa que pode permitir a caracterização do padrão das criptas de Lieberkühn, contornando técnicas de coloração. OBJETIVO: Analisar, quantificar e caracterizar a morfologia do padrão das criptas medindo os parâmetros morfológicos e descritores de forma, através do processamento e análise de imagens de microscopia de autofluorescência de campo de Widefield obtidas em amostras de tecido de cólon fresco a partir de um modelo murino de câncer colorretal. MÉTODOS: Camundongos machos adultos BALB/cCmedc (n=27), variando de 20 a 30 g, foram distribuídos aleatoriamente em quatro e cinco grupos de animais tratados e de controle. As amostras de cólon foram coletadas no dia zero e na 4ª, 8ª, 16ª e 20ª semanas após o tratamento com azoxometano. Segmentação bidimensional (2D), quantificação e caracterização morfológica do padrão das criptas por processamento de imagem aplicados utilizando programação macro de FIJI. RESULTADOS: O tipo I é a morfologia da cripta prevalente entre 53% e 81% semanas do grupo controle. Os tipos III-L e III-S foram detectados em porcentagens reduzidas. A morfologia do tipo I entre os 33% e 56% foi constatada como a predominante para as 4ª, 8ª e 20ª semanas de grupos tratados, seguidos pelo tipo III-L. Para a 16ª semana, os 39% dos padrões das criptas foram caracterizados como tipo III-L, seguidos pelo tipo I. Além disso, os tipos de padrão como IV, III-S e II também foram encontrados principalmente nessa ordem para quase todas as semanas tratadas. CONCLUSÃO: Estes resultados preliminares podem ser considerados um avanço na caracterização bidimensional da cripta como um processamento integral da imagem, comparando-se ao procedimento convencional; demora-se alguns segundos a mais para quantificar e caracterizar pontos não-patológicos, bem como para estimar estágios patológicos precoces do câncer colorretal.

Structural-dependent effects of dietary fibers in colon cancer: Focus on dietary fiber naturally changed by the papaya ripening / Efeitos estrutura-dependente das fibras alimentares no câncer de cólon: foco na fibra alimentar naturalmente modificada durante o amadurecimento do mamão papaia

ietary fiber (DF) consumption is related with several healthy benefits such as the decreasing risk of colon cancer development. The DF is not digested by the digestive enzymes and reach to colon where is fermented by the colonic microbiota. The fermentation process releases metabolites as short chain fatty acids (SCFA) such as butyrate, propionate and acetate. Besides the fermentation process, the DF can directly interact with intestinal epithelial cells inducing mechanism that can also be related with the associated DF consumption benefits. The lack of information regarding DF and colon cancer are due to the complexity of both the cancer and the DF structure. The papayas DF are derived from the fruit cell wall, and they are probably naturally modified during ripening through a massive polysaccharide hydrolysis, because papayas show a very fast pulp softening. Due to the lack of information about DF and their beneficial effects to human health as well as the possibility of the natural papaya ripening to modifying the DF presented in the fruit pulp, the present thesis had as the primary objectives: 1) to evaluate how the cell-wall degrading enzymes affect the fruit cell wall solubilization and molecular weight; 2) to investigate the direct effects of the papaya pectin derived from unripe to ripe papayas in cancer cell lines, in galectin-3 interaction and in HEK cells expressing pattern recognition receptors (PRR); 3) to evaluate the human colonic in vitro fermentation using DF from unripe and ripe papayas as substrates; 4) to conduct an in vivo experiment using rats with pre-neoplastic colon lesions while receiving a diet with DF from unripe and ripe papayas. The endopolygalacturonases were the main enzymes acting on the solubilizing papaya cell wall pectin affecting both the papaya firmness and pectin structure. Overall, the papayas DF showed a ripening dependent structureeffects. In the cancer cell lines experiments, the ripe papayas pectin showed a more pronounced effects in inducing cancer cell death, inhibiting cancer cells migration and aggregation, activating PRR as toll-like receptors and inhibiting the pro-metastatic protein galectin-3. The DF from papayas also showed different aspects in colonic in vitro fermentation regarding the DF utilization by the bacteria and the bacteria abundance profile. Lastly, the animals receiving the diet with the DF from ripe papayas had less aberrant crypt foci in colon than the animals that received the DF from unripe papayas or cellulose (AIN-93G DF). Therefore, the study of papaya DF was carried out both during papaya ripening and its biological effects in vitro and in vivo, generating unprecedented results relating the endogenous biochemical changes of the fruits during maturation with the possible beneficial effects of their ingestion for health human

O consumo de fibras alimentares (FA) está relacionado com vários benefícios à saúde como a diminuição no risco do desenvolvimento de câncer de cólon. A FA não é digerida pelas enzimas digestivas do trato gastrointestinal sendo fermentada pela microbiota intestinal do cólon. Como subproduto do processo de fermentação há a liberação de ácidos graxos de cadeia curta (SCFA) - como o butirato, o propionato e o acetato. Além do processo de fermentação, a FA pode interagir diretamente com as células epiteliais do intestino, induzindo mecanismos que também podem estar relacionados com os benefícios associados ao consumo de FA. A falta de informação sobre a FA e o câncer de cólon é, em partes, devido à complexidade de ambos, tanto do câncer quanto da estrutura da FA. As FA do mamão papaia são derivadas da parede celular da fruta apresentando diferentes estruturas dependendo do ponto de amadurecimento do fruto. Esse fato ocorre, pois, durante o amadurecimento do mamão papaia, existe uma extensa hidrólise dos polissacarídeos presentes na parede celular, diminuindo rapidamente a firmeza da polpa do fruto. Devido à falta de informações sobre FA e seus efeitos benéficos à saúde humana que são dependentes da sua estrutura, bem como a possibilidade do amadurecimento do mamão papaia naturalmente modificar as FA presentes na polpa dos frutos, a presente tese teve como principais objetivos: 1) avaliar como as enzimas que degradam a parede celular do mamão papaia afetam a solubilização e o peso molecular da parede celular do fruto; 2) investigar os efeitos diretos da pectina derivada de mamões verdes e maduros em linhagens de células de câncer, na interação com a galectina-3, e em células do tipo HEK que expressam receptores de reconhecimento de padrões (RRP); 3) avaliar a fermentação colônica humana in vitro utilizando as FA de mamões verdes e maduros; 4) avaliar em ratos com lesões pré-neoplásicas no cólon o efeito do consumo de ração com ou sem FA de mamões papaias verdes e maduros. As endopoligalacturonases foram relacionadas como as principais enzimas que atuam solubilizando a pectina da parede celular do mamão, afetando tanto a firmeza da polpa do fruto quanto a solubilização da pectina durante o amadurecimento. De modo geral, as FA dos mamões exerceram um efeito estruturadependente de acordo com a maturação do fruto. Nos experimentos utilizando linhagens de células de câncer, a pectina do mamão papaia maduro apresentou efeitos mais pronunciados na indução da morte e na inibição da migração e da agregação das células, bem como ativando os RRP, como por exemplo, os receptores do tipo toll-like, além de inibir a proteína pró-metastática galectina-3. As FA dos mamões também apresentaram diferentes resultados na fermentação colônica in vitro quanto à utilização das FA pelas bactérias do intestino, e também no perfil de crescimento dessas bactérias. Por fim, os animais que receberam a dieta com as FA dos mamões maduros apresentaram menor incidência de focos de criptas aberrantes do que os animais que receberam as FA provenientes de mamões verdes ou de celulose (FA da ração AIN-93G). Portanto, o estudo das FA dos mamões foi efetuado tanto durante o amadurecimento dos mamões quanto dos seus efeitos biológicos in vitro e in vivo, tendo gerado resultados inéditos relacionando as alterações bioquímicas endógenas dos frutos durante o amadurecimento com os possíveis efeitos benéficos da sua ingestão para a saúde humana

Protective effects and mechanism of keratinocyte growth factor combined with hypoxia inducible factor-1α on intestinal crypt epithelial cells of rats with hypoxia stress / 中华烧伤杂志

Objective@#To investigate the protective effects and mechanism of keratinocyte growth factor (KGF) combined with hypoxia inducible factor-1α (HIF-1α) on intestinal crypt epithelial cells (IEC-6) of rats with hypoxia stress.@*Methods@#(1) The routinely cultured IEC-6 of rats were collected and divided into normoxia blank group, normoxia KGF group, normoxia HIF-1α group, and normoxia combine group, according to the random number table, and then the previous mediums were respectively replaced with dulbecco′s modified eagle medium (DMEM), medium with 0.5 ng/mL KGF, medium with 10.0 ng/mL HIF-1α, and medium with 0.5 ng/mL KGF and 30.0 ng/mL HIF-1α. And the cells were cultured in cell incubator with oxygen volume fraction of 21% for 24 hours. (2) Another batch of routinely cultured IEC-6 were collected and divided into normoxia control group, hypoxia control group, hypoxia KGF group, hypoxia HIF-1α group, and hypoxia combine group, according to the random number table. The previous mediums were replaced with DMEM, DMEM, medium with 0.5 ng/mL KGF, medium with 10.0 ng/mL HIF-1α, and medium with 0.5 ng/mL KGF and 30.0 ng/mL HIF-1α respectively. And then, the cells in normoxia control group were cultured routinely for 24 hours, and cells in the other 4 groups were cultured in cells incubator of 3 gases, with oxygen volume fraction of 5% for 24 hours. Cells cultured in normoxic and hypoxic incubators were collected, with 3 samples in each group, and morphological changes of cells were observed with optical microscope. Cells cultured in normoxic and hypoxic incubators were collected, with 3 samples in each group, and survival rates of cells were detected by cell count kit 8. Cells in normoxia control group and cells cultured in hypoxic incubator were collected, with 3 samples in each group. The cell cycle changes and apoptosis rates were detected by flow cytometer, the content of adenosine triphosphate (ATP) was detected by ultraviolet spectrophotometer, and protein expression of p53 was detected by Western blotting. Data were processed with one-way analysis of variance and least significant difference test.@*Results@#(1) After being cultured for 24 h, cells cultured in normoxic incubator grew well with oval or round shapes and clear cytoplasm, and cells cultured in hypoxic incubator showed irregular shapes such as fusiform or starlike shape, with black particle in cytoplasm. (2) After being cultured for 24 h, cell survival rates of normoxia blank group, normoxia KGF group, normoxia HIF-1α group, and normoxia combine group were (107.4±8.7)%, (109.8±2.9)%, (115.8±7.4)%, and (112.8±10.6)% respectively. There was no significantly statistical difference in general comparison of cell survival rates among the above groups (F=0.685, P=0.586). After being cultured for 24 h, cell survival rates of hypoxia control group, hypoxia KGF group, hypoxia HIF-1α group, and hypoxia combine group were (35.1±4.6)%, (52.9±6.8)%, (56.2±3.1)%, and (71.2±9.6)% respectively, which were significantly lower than (106.3±12.3)% of normoxia control group (P<0.001). Survival rates of cells in hypoxia KGF group, hypoxia HIF-1α group, and hypoxia combine group were significantly higher than the rate of cells in hypoxia control group (P=0.023, 0.009, <0.001). Survival rate of cells in hypoxia combine group was significantly higher than the rates of cells in hypoxia KGF group and hypoxia HIF-1α group (P=0.017, 0.045). (3) After being cultured for 24 h, percentage of cells in G1 phase in hypoxia control group was significantly higher than that of cells in normoxia control group (P=0.030), percentages of cells in S phase in hypoxia control group, hypoxia KGF group, and hypoxia HIF-1α group were obviously lower than the percentage of cells in normoxia control group (P=0.020, 0.031, 0.026), and percentages of cells in different phases in other groups were close to those of cells in normoxia control group (P=0.516, 0.107, 0.052, 0.985, 0.637, 0.465, 0.314, 0.591). After being cultured for 24 h, percentages of cells in G1 phase in hypoxia control group, hypoxia KGF group, and hypoxia HIF-1α group were obviously higher than the percentage of cells in hypoxia combine group (P=0.001, 0.030, 0.014), and percentages of cells in S phase in the above 3 groups were obviously lower than the percentage of cells in hypoxia combine group (P=0.001, 0.012, 0.010). (4) After being cultured for 24 h, compared with that of cells in normoxia control group, apoptosis rate of cells in hypoxia control group obviously increased (P=0.018), and apoptosis rate of cells in hypoxia combine group obviously decreased (P=0.008). After being cultured for 24 h, compared with that of cells in hypoxia control group, apoptosis rates of cells in hypoxia KGF group and hypoxia combine group obviously decreased (P=0.004, 0.001). Apoptosis rate of cells in hypoxia combine group was obviously lower than those of cells in hypoxia KGF group and hypoxia HIF-1α group (P=0.032, 0.002). (5) After being cultured for 24 h, compared with that of cells in normoxia control group, the content of ATP of cells in hypoxia combine group changed unobviously (P=0.209), and content of ATP of cells in the other groups obviously decreased (P= <0.001, 0.001, 0.002). Content of ATP of cells in hypoxia HIF-1α group and hypoxia combine group was obviously higher than that of cells in hypoxia control group (P=0.044, 0.001). Content of ATP of cells in hypoxia combine group was obviously higher than that of cells in hypoxia KGF group and hypoxia HIF-1α group (P=0.011, 0.020). (6) After being cultured for 24 h, protein expressions of p53 of cells in hypoxia control group, hypoxia KGF group, and hypoxia HIF-1α group were obviously higher than that of cells in normoxia control group (P<0.001), and protein expression of p53 of cells in hypoxia combine group was obviously lower than those of cells in hypoxia control group, hypoxia KGF group, and hypoxia HIF-1α group (P=0.001, 0.001, 0.002).@*Conclusions@#KGF combined with HIF-1α have significant protective effects on IEC-6 of rats with hypoxia stress, and can improve its survival in hypoxic environment by inhibiting cell cycle arrest, reducing the level of apoptosis, and increasing level of energy metabolism.

Advances in research on treatment of aberrant crypt foci / 实用肿瘤学杂志

Aberrant crypt foci(ACF)is highly similar to colorectal cancer(CRC)in phenotypic and molecular changes,and is positively correlated with the risk of CRC. It is considered as precancerous lesion of CRC. Many compounds have therapeutic effects on ACF,inhibiting their occurrence and malignant transformation,which can be regarded as a preventive effect on CRC. In this paper,we will review the treatment of ACF by various compounds in different ways in recent years.

Effect of cottonseed oil and ferrous sulfate in broiler diets on intestinal morphometry at 42 days of age / Efeito de óleo de semente de algodão e sulfato ferroso em dietas de aves na morfometria intestinal aos 42 dias de idade

The aim of the study was to evaluate the intestinal morphometry of broilers at 42 days of age fed diets containing different levels of cottonseed oil, with and without addition of ferrous sulfate. A total of 560 male Ross chickens were used in a completely randomized design with a 4×2 factorial arrangement and five replications with 14 birds. The cottonseed oil levels tested were 0%, 2%, 4%, and 6%. At 42 days of age, one bird per replicate was collected for morphometric. Analyzed variables were villus height, crypt length and muscle wall thickness. The statistical analysis was performed using SAS, P=0.05. In the duodenum and ileum, there was no significant effect of interaction between the treatments. In the jejunum, only the length of the crypt, the treatment with cottonseed oil 2%, more ferrous sulfate resulted in lower values; however, the same oil content without ferrous sulfate led to a greater length, which was also observed in the treatment with 4% oil plus ferrous sulfate. Inclusion of cottonseed oil in broiler diets at 42 days of age does not impair their intestinal morphology. Supplementation with ferrous sulfate at these levels is unnecessary, since no improvement was observed in the intestinal morphometric.(AU)

O objetivo de estudo foi avaliar a morfometria intestinal de aves aos 42 dias de vida alimentadas com dietas contendo níveis variados de óleo de semente de algodão com e sem adição de sulfato ferroso. Um total de 560 machos de aves Ross foram usadas em estudo randomizado com amostras 4x2 e 5 replicações com 14 aves. Os níveis de óleo de semente de algodão testados foram 0%, 2%, 4%, e 6%. Aos 42 dias de idade, um pássaro por replica foi coletado para morfometria. As variáveis analisadas foram altura de villus, comprimento de cripta, e espessura de parede muscular. A análise estatística foi realizada com SAS, P=0,05. Não houve efeito significativo de interação entre tratamentos no duodeno ou íleo. No jejuno, apenas o comprimento da cripta, no tratamento com óleo de semente de algodão a 2% com mais sulfato ferroso, apresentou valor mais baixo. No entanto, o mesmo valor de óleo sem sulfato ferroso resultou em maior comprimento, resultado percebido também com 4% de óleo. Inclusão de óleo de semente de algodão em dietas de aves aos 42 dias de idade não causa danos à morfologia intestinal. Suplementação com sulfato ferroso não é necessário, já que não houve melhora em morfometria intestinal.

Topographic, histological and molecular study of aberrant crypt foci identified in human colon in different clinical groups

BACKGROUND/AIMS: Aberrant crypt foci (ACF) are early microscopic lesions of the colonic mucosa, which can be detected by magnified chromoendoscopy. Herein, we have investigated whether ACF identified in different clinical groups can be differentiated based on their characteristics. METHODS: Macroscopically unremarkable mucosal flaps were collected from 270 fresh colectomies and divided into 3 clinical groups: colorectal carcinoma (group A), disease controls having known pre-neoplastic potential (group Bc), and disease controls without risk of carcinoma development (group Bn). Topographic and histologic analysis, immunohistochemistry, and molecular studies (high-resolution melt curve analysis, real-time polymerase chain reaction, and Sanger sequencing) were conducted for certain neoplasia-associated markers. RESULTS: ACF were seen in 107 cases, out of which 72 were left colonic ACF and 35 right colonic ACF (67.2% vs. 32.7%, P=0.02). The overall density of left colonic ACF was 0.97/cm, which was greater than the right colonic ACF density of 0.81/cm. Hypercrinia was present in 41 out of 72 left colonic ACF and in 14 out of 35 right colonic ACF (P=0.01). Immunohistochemical expression of p53 was also greater in left colonic ACF than in right colonic ACF (60.5% vs. 38.2%, P=0.03). However, ACF identified among the 3 clinical groups did not show any distinguishing topographic, histological, or genetic changes. CONCLUSIONS: Left colonic ACF appear to be high-risk based on their morphological and prototypic tumor marker signature. ACF identified in different clinical groups do not show significant genotypic or topographic differences. Further detailed genetic studies are required to elucidate them further.

Comentario del caso Ana / Commentary of the case Ana

En el comentario del caso se analiza una viñeta clínica que se inicia con un estado de adhesividad violenta entre una adolescente, Ana, y su madre. Para la madre de Ana el mundo es enigmáticamente peligroso y no tolera la separación de su hija. A partir de las manifestaciones de los integrantes de la familia, y el develamiento de la historia familiar, se hacen presentes los pensamientos encriptados, transmitidos a través de las generaciones. Los elementos traumáticos, indecibles en la abuela de Ana, innombrables para su madre, adquieren figurabilidad gracias a que Ana, en el trabajo con su terapeuta comienza a ligarlos con su historia.

A clinical vignette that begins with a state of violent adhesiveness between a teenager, Ana, and her mother is analyzed. For Ana's mother, the world is enigmatically dangerous and she does not tolerate being separated from her daughter. From the manifestations of the members of the family, and the unveiling of the family story, the encrypted thoughts transmitted through the generations are present. The trauma, unsaid for Ana's grandmother, and unspeakable for her mother, acquires figurability for Ana, that working with her therapist, begins to link it with her story.

Dans le commentaire du cas, une vignette clinique qui commence par un état d'adhésivité violente entre une adolescente, Ana, et sa mère est analysée. Pour la mère d'Ana, le monde est énigmatiquement dangereux et elle ne tolère pas la séparation de sa fille. Depuis les récits des membres de la famille et le dévoilement de l'histoire familiale, des pensées cryptées transmises à travers les générations émergent. Les éléments traumatiques, indicibles par la grand-mère d'Ana, innommables pour sa mère, acquièrent de la figurabilité grâce à Ana, qui dans le travail avec sa thérapeute, commence à les relier à son histoire.

Protective effects and possible mechanism of 12-O-tetradecanoylophorbol-13-decano-ate against acute intestinal radiation injury of mice / 军事医学

Objective To investigate the effect of 12-O-tetradecanoylophorbol-13-decanoate(TPD)on protection against acute intestinal radiation injury of mice and the possible mechanism.Methods Twenty female BALB/c mice aged 6-8 weeks were divided by random number table method into the control group and TPD groups(25,50,and 100 μg/kg). A radiation-damaged model of mice was irradiated by 10 Gy 60Co γ-rays,while the TPD groups were pretreated for 3 d with caudal vein injection before irradiation.The survival time of 20 days and the number of crypts at 3.5 days after irradiation were detected.Rat intestinal epithelial cells(IEC-6)were treated with 1 nmol/L TPD for 12 h before irradiation with 10 Gy 60Co γ-rays,and CCK-8 was used to detect the capability of cell proliferation at 0,1,2,3 and 4 d after irradiation. Results The mice in the control group survived for an average of 4.2 days,compared to 10 days in the optimal TPD group (100 μg/kg).The average number of crypts in the control group and the best TPD group was 11.0 ±1.3 and 35.1 ±1.9 respectively.The proliferation activity of IEC-6 was measured for four consecutive days.The average D value of the TPD groups was significantly higher than that of control.Conclusion TPD has a protective effect against acute intestinal radiation injury, and its protective mechanism may be achieved by promoting intestinal crypt cell proliferation and increasing the number of crypts in the intestine.

Mesenchymal stem cells transplantation promote the structural repairation of mice intestinal injury induced by radiation / 重庆医学

Objective To study the effect of human umbilical cord mesenchymal stem cells (hMSCs) on the survival rate of mice with radiation-induced intestinal injury and its effect on the repair of intestinal damage.Methods Adult C57BL/6J mice were selected and divided into control group and abdomen irradiation + PBS(IR + PBS) group and abdominal irradiation+hMSCs transplantation treatment (IR+hMSCs)group.Intestinal injury model was induced by abdominal irradiation at 13 Gy of T ray and the 30-day survival rate was recorded.On the 3.5th and 5.0th day after irradiation,HE staining was used to observe changes in the villus structure and thc number of intestinal crypts in each segment of the small intestine (duodenum,jejunum,and ileum).Results The survival rate of IR+hMSCs group was significantly higher than that of IR+PBS group (P＜0.05).On the 3.5th day after 13 Gy of abdomen irradiation,the duodenum,cavities and ileum villus in IR+PBS group were all fractured,the length was shortened,and the numbers were sparse when compared with the control group,while the hMSCs reduced the intestinal damage.On the 5.0th day after irradiation,the structural integrity of the small intestine in IR+PBS group was repaired slightly,but it still more serious than that of IR+hMSCs group (P＜0.01).The amount of intestine crypts were significantly higher in IR+hMSCs group than that of IR+PBS group,but were significantly lower than that of control group (P＜0.01) at 3.5th day and 5.0th day after radiation.Conclusion Transplantation of hMSCs can improve the survival rate and promote the repairation of intestinal injury induced by radiation in mice.MSCs are hopeful to be used on the treatment of radiation-induced intestinal injury.

Effect of MIF antibody on aberrant crypt foci and colorectal carcinoma in rat / 重庆医学

Objective To study the effect of macrophage migration inhibitory factor(MIF) antibody on the rat colonic aberrant crypt foci(ACF) induced by 1,2-dimethylhydrazine(DMH),carcinoma number and expression of MIF in rat colonic carcinogenesis.Methods Rat colonic carcinogenesis model was induced by DMH.In this model,the inhibitory effect of MIF antibody on the number of ACF and carcinoma was observed.ELISA and immunohistochemical staining were adopted to investigate the effect of MIF antibody in early cancerative intestinal mucosa and MIF expression after cancer formation.Results The number of ACF and carcinoma was significantly inhibited by MIF antibody intervention(P＜ 0.01).The expression of MIF in the colonic carcinoma model was significantly higher than that in the pre-carcinoma ACF model(P＜0.01).Applying MIF antibody could significantly inhibit the expression of MIF in both rat colonic ACF and colonic carcinoma model.Conclusion MIF antibody can significantly inhibit the rat colonic mucosal carcinogenesis,which may be related with inhibiting number of ACF and expression of MIF.MIF antibody may be expected to become a new target spot of precaution and treatment of colonic carcinoma.

Negative Histology after Endoscopic Resection: A Pathologist's Aspect / 대한Helicobacter연구학회지

Endoscopic biopsy is the most popular diagnostic procedure for gastric epithelial neoplasm. However, discrepancy between biopsy and endoscopic resection diagnosis occasionally occurs and leads to an unnecessary procedure. A negative pathologic diagnosis following endoscopic resection is particularly problematic. In this context, we reviewed factors that may be associated with a diagnostic discrepancy between endoscopic biopsy and endoscopic resection, with focus on crypt dysplasia.

Transgeracionalidade: sobre silêncios, criptas, fantasmas e outros destinos / Transgeneracionalidad: sobre silencios, criptas, fantasmas y otros destinos / Transgenerational identity ("transgenerationality"): about silences, crypts, phantoms, and other destinies / Transgénérationalité: sur les silences, les cryptes, les fantasmes et d'autres destins

L'auteur présente les voies qui peuvent conduire aux héritages transgénérationnelles au sein de plusieurs familles à partir de traumas, de secrets, de hontes et de deuils qui ne sont pas travaillés, lorsque ceux-ci, au moyen du silence et de la coupure employés comme moyens de défense, résultent dans la formation de cryptes et de fantasmes. Elle présente aussi les idées de transmission intergénérationnelle et transgénérationnelle

The author presents paths which start from situations of trauma, secret, shame, not elaborated mourning, and may lead to transgenerational heritages within several families. It happens when silence and splitting, used as defense mechanisms, end up creating crypts and phantoms. This paper also presents ideas of intergenerational and transgenerational transmission.

La autora presenta los caminos que pueden conducir a las herencias transgeneracionales en el seno de muchas familias a partir de traumas, secretos, vergüenzas, duelos no elaborados, cuando, a través del silencio y de la cisión, utilizados como defensas, producen criptas y fantasmas. También presenta un cuadro sinóptico con las transmisiones intergeneracionales y transgeneracionales.

A autora apresenta os caminhos que podem conduzir às heranças transgeracionais no seio de muitas famílias a partir de traumas, segredos, vergonhas, lutos não elaborados, quando, através do silêncio e da cisão, usados como defesas, resultam na formação de criptas e fantasmas. Também apresenta as ideias de transmissão intergeracional e transgeracional.

Primary cultures of mouse small intestinal epithelial cells using the dissociating enzyme type I collagenase and hyaluronidase

The epithelium is a highly dynamic system, which plays a crucial role in the homeostasis of the intestinal tract. However, studies on the physiological and pathophysiological functions of intestinal epithelial cells (IECs) have been hampered due to lack of normal epithelial cell models. In the present study, we established a reproducible method for primary culture of mouse IECs, which were isolated from the viable small intestinal crypts of murine fetuses (on embryonic day 19), using type I collagenase and hyaluronidase in a short span of time (≤20 min). With this method, continuously growing mouse IECs, which can be subcultured over a number of passages, were obtained. The obtained cell lines formed a tight cobblestone-like arrangement, displayed long and slender microvilli, expressed characteristic markers (cytokeratin 18 and Notch-1), and generated increasing transepithelial electrical resistance and low paracellular permeability during in vitro culture. The cells also had enzymatic activities of alkaline phosphatase and sucrase-isomaltase, and secreted various cytokines (IL-1β, IL-6, IL-8, and monocyte chemoattractant protein-1), responding to the stimulation of Escherichia coli. These results show that the primary-cultured mouse IECs obtained by the method established here had the morphological and immunological characteristics of IECs. This culture system can be a beneficial in vitro model for studies on mucosal immunology and toxicology.

Effect of Yiqi Jianpi Fang on Aberrant Crypt Foci Production in Carcinogenesis Wistar Rat Colorectal Cancer Model and Investigation on the Related Mechanism / 现代生物医学进展

Objective:To investigate the inhibitory effect and the related mechanism of Yiqi Jianpi Fang on aberrant crypt focus (ACF) in Wister rat colorectal cancer model.Methods:50 Wistar rats were randomly divided into 5 groups:TCM low dose group(TCM solution diluted 3 times,5 mL/kg daily gastric volume),TCM middle dose group (5 mL/kg daily gastric volume),TCM high dose group (15 mL/kg daily gastric volume),model group,normal group.With 10 rats in each group.The colorectal tissues were observed under microscope after methylene blue staining by immunohistochemistry method.Results:Compared with the model group,the number of ACF and large ACF in each TCM group were decreased (P＜0.05),and the number of ACF in the TCM middle dose group reduced most obvious,and the difference was statistically significant (P＜0.05).The proliferation indexs of PCNA in intestinal gland cells 24 h and 48 h after modeling in the TCM groups were higher than those in the model group,and the difference was statistically significant(P＜0.05).The apoptosis index of intestinal gland cells 24 h and 48 h after modeling in the TCM groups were higher than those in the model group,and the difference was statistically significant(P＜0.05).The ectopic expression of β-catenin in TCM groups were lower than that in the model group,and it was highest in the high dose group,than was the low dose group,and the middle group was lowest(P＜0.05).The expression of MMP-7 in TCM groups were lower than that in the model group,and it was highest in the low dose group,than was the high dose group,and the middle dose group was lowest (P＜0.05).Conclusion:Yiqi Jianpi Fang can significantly reduce the number of ACF in Wister rats,inhibit the activation of Wnt signaling in colorectal cancer,reduce the incidence of colorectal cancer,and have a certain preventive effect.

Uma história atual (1768-1922): retratos de um segredo / A recent and current story (1768-1922): portraits of a secret / Una historia actual (1768-1922): retratos de un secreto

Partindo do pressuposto de que a transmissão psíquica transgeracional é constitutiva do psiquismo do indivíduo, o presente artigo tem como objetivo discutir sobre os silêncios transgeracionais, sobre a falta da palavra e sobre o legado entre gerações, que, sendo silenciado por vergonha ou pelo excesso de dor e não podendo ser transmitido aos descendentes, produz consequências significativas nas gerações seguintes. O trabalho baseia-se na história de uma mulher, terceira geração de imigrantes que se refugiaram no Brasil.

In this paper, the author assumes that transgenerational psychic transmission is constitutive of the individual's psyche. Starting from that premise, the author's purpose is to discuss transgenerational silences, the lack of word, and the legacy between generations. As a result of being silenced either by shame or excessive pain, and for not being able to be transmitted to descendants, that legacy has significant effects across the following generations. This paper is based on a woman's story. That woman belongs to the third generation of immigrants who came to Brazil.

Partiendo de la suposición de que la transmisión psíquica transgeneracional es constitutiva de la psique individual, este artículo tiene como objetivo discutir sobre los silencios transgeneracionales, sobre la falta de la palabra, y sobre el legado entre generaciones que fue silenciado, por vergüenza o por exceso de dolor, y que no pudo transmitirse a los descendientes, lo que genera consecuencias importantes en las siguientes generaciones. El trabajo se basa en la historia de una mujer, tercera generación de inmigrantes que se refugiaron en Brasil.

Dietas suplementadas com óleos essenciais não promove alterações na morfometria de vilos e criptas do duodeno de frangos de corte / Diets supplemented with essential oils do not promote alterations in villous morphometry and duodenum crypt of broilers / Dietas suplementadas con aceites esenciales no promove cambios en morfometría de las vellosidades y criptas del duodeno de pollos de engorda

Devido à necessidade da substituição dos antimicrobianos como melhoradores de desempenho por aditivos alternativos, inúmeros produtos estão em estudo. Dentre esses estão os óleos essenciais que apresentam características antimicrobianas, contribuindo para manutenção da integridade intestinal e consequentemente melhora no desempenho, garantindo a segurança alimentar, evitando a resistência cruzada, bem como a contaminação do meio ambiente. Dessa forma, objetivou-se avaliar os efeitos da inclusão de uma mistura de óleos essenciais extraídos de orégano, alho, limão, alecrim, timo, eucalipto e laranja doce, na dosagem de 300 mL para 500 L de água de bebida, sobre a morfometria da mucosa do duodeno de frangos de corte. Foram avaliados quatro aviários tratados com óleos essenciais na ração e quatro aviários controle, durante o período de um a sete dias de idade, contendo pintainhos da linhagem Cobb. Para a análise morfométrica, cinco aves de cada aviário (total de 40 aves) foram mortas, utilizando-se de protocolo anestésico, medicação pré-anestésica de xilazina 4mg/Kg IM. e após solução de tiopental na dose de 25 mg/Kg IV. Após eutanásia, foram colhidas amostras de material do intestino delgado, correspondente ao segmento duodenal, para realização das medições de altura de vilo e profundidade de cripta. Não foram verificados efeitos sobre a altura de vilo e profundidade de cripta nos animais tratados com óleos essenciais (P>0,05). Nas condições em que foram realizados o experimento, pode-se concluir que os óleos essenciais não alteraram a altura de vilo e a profundidade de cripta do duodeno.

Due to the need for replacement of antimicrobials as performance enhancers to alternative additives, several products are being studied. Among them, essential oils that exhibit antimicrobial properties are being analyzed, which contribute to the maintenance of intestinal integrity and consequently improving performance, assuring food safety, preventing cross-resistance, as well as contamination of the environment. Thus, the objective of this study is to evaluate the effects of adding a mixture of essential oils from oregano, garlic, lemon, rosemary, thyme, eucalyptus and sweet orange, at a dosage of 300 mL for 500 L of drinking water, on the morphometry of duodenal mucosa of broilers. Four poultry aviaries treated with essential oils in the diet and four control poultry aviaries were evaluated during the period from one to seven day old, containing Cobb chicks. For the morphometric analysis, five birds in each aviary (total of 40 birds) were euthanized using anesthetic protocol premedication with xylazine 4mg/kg IM) and after, a solution of thiopental (25 mg /Kg IV). After euthanasia, material samples were collected from the small intestine (duodenum) for measuring villus height and crypt depth. No effects on villus height and crypt depth could be found in the animals treated with essential oils (P> 0:05). Under the experimental conditions, it can be concluded that essential oils did not alter villus height and crypt depth of the duodenum.

Debido a la necesidad de reemplazo de antimicrobianos como mejoradores de desempeño por aditivos alternativos, innúmeros productos están siendo estudiados. Entre esos están los aceites esenciales que presentan características antimicrobianas, lo que contribuye al mantenimiento de la integridad intestinal y por consiguiente mejora el rendimiento, por garantizar la seguridad alimentaria, evitando resistencia cruzada, así como la contaminación del medio ambiente. Por lo tanto, el objetivo ha sido evaluar los efectos de la adición de una mezcla de aceites esenciales extraídos del orégano, ajo, limón, romero, tomillo, eucalipto y naranja dulce, a una dosis de 300 ml para 500 litros de agua potable, sobre la morfometría de la mucosa duodenal de pollos de engorde. Se han evaluado cuatro aviarios tratados con aceites esenciales en el pienso y cuatro aviarios control, durante el período de uno a siete días de edad, conteniendo pollitos Cobb. Para el análisis morfométrico, cinco aves de cada aviario (total de 40 aves) fueron sacrificadas, utilizándose de protocolo anestésico, medicación pre anestésica de xilazina 4 mg /Kg IM., y después tiopental solución a una dosis de 25 mg /Kg IV. Después de la eutanasia, se ha recogido muestras de material del intestino delgado, que corresponde al segmento duodenal, para llevar a cabo las mediciones de la altura de las vellosidades y profundidad de la cripta. No se ha verificado efectos sobre la altura de las vellosidades y la profundidad de la cripta en los animales tratados con aceites esenciales (P> 0,05). En las condiciones en que se han realizado el experimento, se puede concluir que los aceites esenciales no alteraron la altura de las vellosidades y la profundidad de la cripta del duodeno.

Extract of the Bark of Bathysa cuspidataAttenuates the Development of Chemically-Induced Preneoplastic Colorectal Lesions in Rats

ABSTRACTThe aim of this study was to investigate the effect of the bark extractBathysa cuspidata on chemically induced preneoplastic colorectal lesions in Wistar rats. Forty male rats were randomly divided into four groups (n = 10 each): saline (control group, oral administration of saline solution 0.9%); dimethylsulfoxide (DMSO, vehicle control), B200 (treated with 200 mg/kg bark extract ofB. cuspidata), and B400 (treated with 400 mg/kg bark extract ofB. cuspidata). Administration of treatments was carried out by the gavage. The animals received four subcutaneous injections of 1,2-dimethylhydrazine (DMH, 40 mg/kg) in the initial two weeks of the experiment to induce preneoplastic colorectal lesions. After 15 weeks, the animals were euthanized and the presence of aberrant crypt foci (ACF), body weight, biochemical analyses, and oxidative stress markers were measured. The extract ofB. cuspidata decreased the levels of superoxide dismutase (SOD), but did not influence the levels of catalase (CAT), malondialdehyde (MDA), nitric oxide or protein carbonyl, compared with the saline group. The animals supplemented with a more concentratedB. cuspidata extract (B400) showed a significant reduction in the number of ACF in all the portions of the intestinal mucosa. The study demonstrated that the bark extract ofB. cuspidata at 400 mg/kg reduced the preneoplastic colorectal lesions in an animal model of colon cancer and that the effect could be dose-dependent.

Interpretation of ileal biopsies.

The ileum is one of the most common sites of intestine to undergo endoscopic biopsy. However, even with the experienced histopathologists, a defi nite diagnosis can be achieved only in 18% cases. Lack of knowledge about proper tissue handling, tissue orientation, overlapping histological fi ndings, and lack of a standard algorithm based approach results in this low diagnostic yield. In this review article, we have tried to discuss these aspects and give a clear picture how to approach the ileal lesions. It would help the surgical pathologists in effectively interpreting the lesions and to identify the common pitfalls.