Model of diabetic keratopathy with high-fat and high-glucose feeding mice / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To discuss the establishment of immediate diabetic keratopathy animal model of C57BL/6 mouse induced by ahigh-fat and high-glucose diet. METHODS: Diabetes mellitus was induced by a high-fat and high-glucose diet in C57BL/6 mouse. 1% rose bengal was stained on the cornea to examine the integrality of the corneal epithelium at 2 ~ 12mo after completion of the model. Corneal epithelial wound healing was observed using a vivo epithelial debridement model which was dyed by sodium fluorescein. Corneal morphology histology was examined by pathological methods. RESULTS: The high-fat and high-glucose diet C57BL/6 mouse in 2mo had showed general symptoms of diabetes: polydipsia, polyphagia, polyuria, weight loss etc. The model had a steady-state high glucose (≥18mmol/L), also the weight was lower compared with normal control mouse. 1% rose bengal corneal staining had dot coloring at 2mo after completion of the model, the stained area and extent were gradually increased with the extension of the duration of diabetes, almost all the cornea was stained at 12mo after completion of the model. With the passage of time into a mold, the cornea epithelial healing time become longer: 2mo was about 40h;3mo was about 120h; 4, 6, 12mo was about 144h;the coloboma were gradually increased at 12mo after completion of the model, then the area was reduced gradually until complete healing, the time was 96~120h, showed repeating phenomenon. CONCLUSION: The mouse were induced by high-fat and high-glucose diet can be used as animal models of diabetic keratopathy: the damage of epithelium for corneal and delay healing on epithelium and other symptoms.

The Feasibility of Using Simulated Targets in the Stomachs of Live Pigs for Full Endoscopic Submucosal Dissection Training

BACKGROUND/AIMS: In endoscopic submucosal dissection (ESD) training, only a flat target lesion can usually be simulated in the normal mucosa. This study aimed to evaluate the feasibility of simulated targets in the stomachs of live pigs for complete training. METHODS: Six trained endoscopists with hands-on experience with ex vivo, isolated pig stomachs were enrolled in this pilot study. An endoscopic banding device was used to create a polyp that was snared, leaving an ulcerated lesion. This simulated target model was used to perform ESD in pigs. The en bloc resection rate, procedure time, complications, quality of resection, and participants' opinions on the simulated targets were compared with the conventional model. RESULTS: En bloc resections were achieved in all six simulated targets and six conventional models. The mean size of the resected specimens was 32.2 mm (range, 20 to 39 mm) in the simulated target group and 23.5 mm (range, 11 to 40 mm) in the conventional group. The target model had a high quality of resection and had a high satisfaction rate for margin identification and correct peripheral marking. CONCLUSIONS: Good identification of the lesion and ease of periphery marking in the target model may improve resection quality.

Animal models as tools to study the pathophysiology of depression

The incidence of depressive illness is high worldwide, and the inadequacy of currently available drug treatments contributes to the significant health burden associated with depression. A basic understanding of the underlying disease processes in depression is lacking; therefore, recreating the disease in animal models is not possible. Popular current models of depression creatively merge ethologically valid behavioral assays with the latest technological advances in molecular biology. Within this context, this study aims to evaluate animal models of depression and determine which has the best face, construct, and predictive validity. These models differ in the degree to which they produce features that resemble a depressive-like state, and models that include stress exposure are widely used. Paradigms that employ acute or sub-chronic stress exposure include learned helplessness, the forced swimming test, the tail suspension test, maternal deprivation, chronic mild stress, and sleep deprivation, to name but a few, all of which employ relatively short-term exposure to inescapable or uncontrollable stress and can reliably detect antidepressant drug response.

Genética molecuaar y biogerontología en la era posgenómica:: un enfoque en las sirtuinas / Molecular Genetics of Aging in the Postgenomic Era:: A Focus on Sirtuins

La forma de estudiar la genética ha progresado notablemente en las últimas décadas. Sus orígenes se remontan al estudio de los caracteres hereditarios, seguido por el descubrimiento de los genes y los cromosomas hasta conocer la estructura del ADN. Este último evento impulsó el desarrollo de la tecnología del ADN recombinante y de la secuenciación masiva y automatizada, los cuales permitieron determinar posterior-mente la anatomía de los genomas. Todos estos descubrimientos han promovido la evolución de la biomedicina hacia las eras genómica y posgenómica en las que el uso de la genética reversa impera sobre la genética básica o directa. Además, surge la genética molecular, la genómica funcional y las diversas tecnologías -ómicas- que en conjunto pretenden comprender de manera integral la función de todos los componentes del genoma y sus productos. La biogerontología, disciplina que estudia los mecanismos biológicos del envejecimiento, es uno de los campos que se han desarrollado notoriamente en los últimos 15 años y refleja los avances científicos de la era posgenómica. Actualmente se han identificado varios gerontogenes y vías moleculares que modifican longevidad y regulan procesos y enfermedades relacionadas con envejecimiento. Dentro de estos genes se encuentran las sirtuinas, una familia de genes conservada evolutivamente que codifica para proteínas con actividad de desacetilasa dependiente de NAD+ y que tienen un papel importante en envejecimiento. En este trabajo revisamos diferentes aproximaciones de genética reversa que se han empleado para identificar algunas de las funciones de estos genes en mamíferos.

The way to study genetics has notably progressed in the last decades. Their origins date back to the study of hereditary features, followed by the discovery of genes and chromosomes up to the knowledge of DNA structure. This last event led to the development of recombinant DNA technology and the massive and automated sequencing, which allowed later to determine the anatomy of genomes. All of these discoveries have pushed the evolution of biomedicine towards the genomic and postgenomic eras, in which the use of reverse genetics prevails over the basic or direct one. Furthermore, it emerges the molecular genetics, the functional genomics and the diverse -omics- technologies that together pretend to understand, in an integrative way, the function of all of the genome components and its products. Biogerontology, discipline that studies the biological mechanisms of aging, is one of the fields that has developed notoriously in the last 15 years and reflects the scientific advances of the postgenomic era. Currently, there have been identified several gerontogenes and molecular pathways that modify and regulate age-related processes and diseases. Among these genes are the sirtuins, an evolutionarily preserved family of genes, which codify for proteins with NAD+ dependent deacetylase activity and that play an important role on aging. In this work, we review different reverse genetics approaches that have been used in order to identify some of the functions of these genes in mammals.

Efeito da utilização de um centrifugado de medula óssea no tratamento de lesão muscular: estudo experimental em coelhos / Effect of use of bone-marrow centrifugate on muscle injury treatment: experimental study on rabbits

OBJETIVO: O objetivo deste estudo foi de avaliar o efeito de um centrifugado de medula óssea na cicatrização de lesão muscular em coelhos. MÉTODOS: Este estudo experimental envolveu a utilização de 15 coelhos machos, adultos, da raça Nova Zelândia Branco. Em cada animal, realizou-se uma lesão transversal na região média do músculo tibial anterior direito, onde se adicionou uma esponja de colágeno absorvível, embebida em um centrifugado de aspirado de medula óssea do osso ilíaco ipsilateral. O membro posterior esquerdo foi utilizado como controle, sendo submetido à mesma lesão; porém, neste caso, adicionando-se somente a esponja de colágeno absorvível. Após 30 dias, os animais foram sacrificados para o estudo da cicatrização muscular. As áreas de cicatrização muscular foram submetidas à análise histológica com histomorfometria, interessando-se a mensuração do número de células musculares em regeneração por micrômetro quadrado e a proporção de fibrose formada. RESULTADOS: O método de centrifugação utilizado neste estudo resultou em uma concentração média de células nucleadas maior que o número destas células nos aspirados originais, sem provocar destruição celular significativa. A adição do centrifugado de medula óssea não resultou em aumento significante do número de células musculares em regeneração em relação ao grupo controle. Também não houve diferença significante na proporção de fibrose formada em relação ao grupo controle. CONCLUSÃO: A administração de centrifugado de medula óssea utilizada neste estudo não favoreceu a cicatrização de lesão muscular em coelhos.

OBJECTIVE: The objective of this study was to evaluate the effect of bone-marrow centrifugate on the healing of muscle injuries in rabbits. METHODS: This experimental study involved use of fifteen adult male New Zealand White rabbits. Each animal received a transverse lesion in the middle of the right tibialis anterior muscle, to which an absorbable collagen sponge, soaked in a centrifugate of bone marrow aspirate from the ipsilateral iliac bone, was added. The left hind limb was used as a control and underwent the same injury, but in this case only the absorbable collagen sponge. Thirty days later, the animals were sacrificed to study the muscle healing. These muscle areas were subjected to histological analysis with histomorphometry, with the aim of measuring the number of muscle cells per square micrometer undergoing regeneration and the proportion of resultant fibrosis. RESULTS: The centrifugation method used in this study resulted in an average concentration of nucleated cells greater than the number of these cells in original aspirates, without causing significant cell destruction. Addition of the bone marrow centrifugate did not result in any significant increase in the number of muscle cells undergoing regeneration, in relation to the control group. There was also no significant difference in the proportion of resultant fibrosis, compared with the control group. CONCLUSION: Administration of the bone marrow centrifugate used in this study did not favor healing of muscle injuries in rabbits.

Programando o software Imagej para a seleção automática de áreas marcadas por imuno-histoquímica e coradas com o cromógeno dab e contra-coradas pela hematoxilina / Programming Imagej software for automatic selection of immunohistochemistry marked areas stained by dab chromogen and counter-stained by hematoxylin

Introdução: A quantificação dos parâmetros de imunomarcação na análise de imagens de imuno-histoquímica depende da adequada delimitação das regiões marcadas, sendo a seleção destas regiões realizada manualmente ou através de métodos computacionais automáticos ou semi-automáticos. Objetivos: Viabilizar um método de seleção automática de áreas marcadas por imuno-histoquímica por meio do software ImageJ, apresentando a programação envolvida e analisando quantitativamente as imagens. Métodos: Para a aplicação e avaliação da programação proposta neste trabalho foram utilizadas imagens de imuno-histoquímica para localização de alfa-actina de músculo liso (α-SMA) provenientes de estudo de modelo animal experimental de fibrose renal em ratos Wistar machos. Foram capturadas imagens de áreas distintas da região do córtex renal contendo regiões positivas coradas pelo 3,3'-Diaminobenzidina (DAB) em uma lâmina com imunomarcação extensa (n=10) e em outra lâmina com Imunomarcação focal (n=10), provenientes de dois animais. A contra-coloração foi feita com hematoxilina. Resultados: A diferença da extensão da imunomarcação foi qualitativamente evidente entre os dois grupos de imagens. As áreas reveladas pelo DAB nas lâminas dos animais I e II foram selecionadas automaticamente pelo programa, ajustadas pelo operador, e quantificadas, apresentando fração de área ocupada de 23,48 ± 6,52% e de 0,10 ± 0,04% respectivamente, valores expressos em média ± desvio padrão com diferença estatística significativa (p < 0,0001).Conclusões: O software ImageJ mostrou-se adequado para a aplicação da seleção automática de áreas marcadas por imuno-histoquímica, embora a intervenção do operador tenha sido necessária para a correta identificação dos pontos de corte do "threshold", supervisionando a correta delimitação das áreas.

Introduction: The immunostaining quantitative parameters of immunohistochemistry image analysis depend on the proper delimitation of marked regions. The selection of those marked regions is carried out manually, or by computed semi-automatic or automatic methods.Objectives: to carry out a feasible automatic method on selection of immunohistochemistry stained areas applying the Image J software (NIH), presenting the computed program and the quantitative analysis of some images.Methods: For application and evaluation of the proposed program of the study, we used immunehistochemistry images of alpha-smooth muscle actin (α-SMA), a marker of epithelial dedifferentiation, coming from an experimental study of renal fibrosis in Wistar male rats. The images have been captured from non-overlapping regions of the renal cortex, comprising positive 3,3'-Diaminobenzidine (DAB)-stained fields of a strong immunostained slide (N=10, animal I) and another slide with focal immune pattern (N=10, animal II), obtained from two different animals. The slides were counter-stained by Hematoxylin.Results: The difference by immunostaning between the two specimens was qualitatively evident. The areas revealed as DAB-positive in animals I and II slides were automatically selected by the proposed program, visually adjusted by the operator, and finally, quantified, resulting in an occupied area fraction of 23.48 ± 6.52% and 0.10 ± 0.04%, respectively, p<0.0001. Conclusion: The software Image J was able to perform a feasible automatic selection of immunohistochemistry DAB stained areas, although the operator intervention has been necessary to identify the correct threshold cut-off points, supervising if the delimitation of marked areas was precise.

Expressão gênica de fatores angiogênicos na placenta de ratas submetidas ao estresse / Expression of the angiogenic factors in the placenta of stressed rats

Objetivos: Avaliar a influencia do estresse sobre os pesos maternos, placentários e fetais, as alterações histológicas placentárias e a expressão gênica dos fatores angiogênicos em ratas prenhes. Métodos: De setembro de 2007 a julho de 2008, foi realizado um estudo experimental do tipo casocontrole, em que 6 ratas prenhes foram submetidas a estímulos estressantes crônicos e agudos enquanto que 6 animais constituíram o grupo controle. No 20º dia de prenhez, todas as ratas foram sacrificadas. Os seguintes dados foram analisados e comparados entre os grupos: a. pesos maternos, placentários e fetais; b. alterações histológicas placentárias; e, c. expressão gênica dos fatores angiogênicos (VEGF-A e PlGF), dos seus receptores (VEGFR-1 e VEGFR-2) e do fator induzido por hipoxia (HIF-1). Resultados: Os pesos maternos, placentários e fetais foram significativamente menores no grupo de ratas prenhes-estressadas em relação ao controle. Histologicamente foram encontradas a presença de núcleos picnóticos no grupo prenhe-estressado. Observou-se expressão gênica significativamente maior de VEGF-A no grupo rata prenhe-estressada assim como redução significante da expressão gênica de PlGF, VEGFR-1, VEGFR-2, quando comparadas ao controle. Não houve alterações entre os grupos para o gene HIF-1. Conclusões: No modelo animal de estresse estudado, observou-se alterações significativas no peso placentário e da expressão gênica dos fatores angiogênicos placentários em ratas submetidas ao estresse.

Objectives: to evaluate the influence of the stress on the maternal, placental and fetal weights, on the histological findings and on the genetic expression of the angiogenic factors in pregnant rats. Methods: From September 2007 to Julie 2008, an experimental case-control study was conduced in which 6 pregnant rats were submitted to chronic and acute stress while six other were considered as controls. In the 20th day of gestation, all animals were sacrified. The following data were evaluated and compared between both groups: a. maternal, placental and fetal weights; b. histological findings; and c. genetic expression of angiogenic factors (VEGF-A and PIGF), receptors (VEGFR-1 and VEGFR-2) and hypoxic-induced factor (HIF- 1). Results: Maternal, placental and fetal weights were statistically smaller in the stressed animals comparing to controls. Histological analysis revealed picnotic nuclei in the placentas of stressed rats. A statistically significant increase in the genetic expression of VEGF-A as well as a reduction of the expression of the PlGF, VEGFR-1, VEGFR-2 were observed in the placentas of the stressed group in comparison to controls. There was no difference of expression of the gene HIF-1 between both groups. Conclusions: In the present animal model of stress, significant alterations in the placental weights, histology and genetic expression of the angiogenic factors among pregnant rats under stress.

Medida da pressão intra-abdominal após colocação de compressas ao redor do fígado: estudo experimental em porcos / Intra-abdominal pressure measurement after sponge placement around the liver: experimental study in pigs

INTRODUÇÃO: A síndrome compartimental abdominal (SCA) é uma complicação frequente da cirurgia de controle de dano, podendo ocorrer em até 33% dos casos. A medida da pressão intra-abdominal para diagnóstico da SCA é avaliada através da pressão intravesical. OBJETIVO: Avaliar, através de modelo experimental, as repercussões da colocação de compressas sobre a pressão intravesical e em diferentes regiões do abdome. MÉTODO: Foram utilizados 10 porcos machos, entre 45 e 60 dias, com peso entre 15 e 18 kg, submetidos a laparotomia exploradora para colocação de compressas e avaliação de medidas de pressão no espaço supra-hepático (Psh), espaço infra-hepático (Pih), espaço subfrênico esquerdo (Psfe), veia cava inferior (Pvci), veia porta (Pvp) e bexiga (Pv). Foi também avaliada a pressão arterial média (PAM) e o débito cardíaco. As diferentes medidas de pressão foram comparadas no mesmo animal com diferentes tipos de fechamento da parede abdominal: bolsa de Bogotá (CCB), total da aponeurose (CCAF), pele (CCPF) e total da pele e aponeurose (CF). Os resultados foram submetidos a análise estatística. RESULTADOS: Não houve diferença (p>0,05) entre as medidas de PAM em todas as fases de fechamento do abdome. A pressão de veia cava inferior e a pressão intravesical foram diferentes (p<0,05) entre os grupos CF e CCAF. Psfe, Psfd e Psh foram diferentes (p<0,05) entre CF e CCB, CF e CCPF e CF e CCAF. Foram diferentes (p<0,05) Pv e Psfd, Pv e Psh e Psfe e Psfd nos grupos CCB, CCPF e CCAF. No grupo CCAF ainda foram diferentes (p<0,05) Pv e Psfe e Psh e Psfd. A avaliação de Pvp mostrou-se diferente (p<0,05) em relação ao grupo controle (CF) nos grupos CCB, CCPF e CCAF, e ainda entre CCB e CCAF. CONCLUSÃO: A pressão intra-abdominal não é uniforme quando são colocadas compressas na cavidade peritoneal. A pressão vesical é menor que em outras regiões da cavidade peritoneal. As pressões não ultrapassaram níveis de Síndrome Compartimental Abdominal.

INTRODUCTION: Abdominal compartment syndrome is a frequent complication of damage control surgery and can occur in approximately 33% of cases. Diagnosis of abdominal compartment syndrome depends on measurement of abdominal pressure which is usually assessed through intravesical pressure. OBJECTIVE: Evaluate the consequences of liver packing with sponges to the intravesical pressure and to pressures in different sites of the abdomen in an animal experimental model. METHODS: 10 adult male pigs, aged 45 to 60 days, weighing 15 to 18 kg, underwent laparotomies for liver packing and evaluation of pressures in suprahepatic space (Psh), infrahepatic space (Pih), left subphrenic space (Psfe), inferior vena cava (Pvci), portal vein (Pvp) and bladder (Pv). Other variables such as mean arterial pressure and cardiac output, were also measured. Different pressure measurements were compared in the same animal with different types of closures of the abdominal wall: Bogota bag (CCB), total closure of aponeurosis (CCAF), skin closure (CCPF) and total skin and aponeurosis closure (CF). Results were analyzed statistically. RESULTS: There was no difference (p>0,05) between mean arterial blood pressure in all phases of abdominal closure. Pressure in inferior vena cava and intravesical pressure were different (p<0,05) in groups CF and CCAF. Psfe, Psfd and Psh were different (p<0,05) between CF vs CCB, CF vs CCPF and CF vs CCAF. Pv vs Psfd, Pv vs Psh and Psfe vs Psfd were different (p<0,05) in groups CCB, CCPF and CCAF. In group CCAF, Pv vs Psfe and Psh vs Psfd were also different (p<0,05). Evaluation of Pvp has shown to be different (p<0,05) when compared to control group (p<0,05) in groups CCB, CCPF and CCAF, and also between CCB and CCAF. CONCLUSION: Intra-abdominal pressure is not uniform when sponges are placed in the abdomen. Intravesical pressure is lower than pressures in other sites of the peritoneal cavity. No pressure measurement reached levels of abdominal compartment...

Efeito da altura da união implante/componente protético sobre os tecidos periimplantares: estudo clínico e radiográfico em cães / The effect of the implant/abutment junction height to the biologic width: clinical and radiographic study in dogs

O objetivo deste estudo foi avaliar as alterações dos tecidos periimplantares, ao redor de implantes instalados em diferentes profundidades em relação à crista óssea, e sob protocolos de reabilitação protética convencional e imediato. Cinco cães receberam trinta implantes, três por hemi-mandíbula (seis por cão), na região dos quatro pré-molares inferiores que foram extraídos 90 dias antes. Cada região foi divida pelos subgrupos convencional (protocolo convencional, instalação da prótese 120 dias após os implantes) e imediato (protocolo imediato, instalação da prótese 1 dia após a cirúrgia de implantes). Os implantes foram inseridos em três profundidades diferentes: ao nível (ao nível do rebordo ósseo), -1 mm (1 mm apicalmente à crista óssea) e -2 mm (2 mm apicalmente à crista óssea). Após acompanhamento por 45 dias da reabilitação instalada foi realizada avaliação clínica e radiográfica e, os dados foram analisados estatisticamente (p< 0.05). A posição do tecido marginal (PTM) não foi alterada pelo protocolo de instalação, e sim pelo nível de inserção da junção implante-conector protético (JIC) em relação a crista óssea (BC). A reabsorção da BC em função do nível de inserção de JIC foi relativamente uniforme para os diferentes grupos. Estes achados sugerem cautela ao posicionar a JIC de implantes apicalmente à BC

The aim of the present study was to evaluate changes in tissues around dental implants inserted in different levels in relation to crestal bone (BC) under early and delayed loading conditions. Thirty implants were inserted in five mongrel dogs were used in this study, from which, the lower premolars were removed, and 90 days after, in one side, three dental implants were inserted (Delayed subgroup, implants installed 120 days before loading) in three different levels in relation to crestal bone (Bone Level, Minus1 and Minus2 groups), and three implants were inserted on remain ridge (Early subgroup, implants were loaded 48 hours after installed). The metallic prostheses were installed in both sides and preserved for 45 days. The clinical and radiographic analyses were performed. Data was submitted to statistics, under a 5% level of significance. The load conditions did not interfered on the position of marginal soft tissues (PTM), those changes evaluated because of the insert level of the implant-abutment junction (JIC) in to BC. The BC resorption was similar in all types of insertion levels of the implants. The present finds suggest caution on insertion of the implant in an apical position level in relation to BC

Study on the model of acute pancreatitis induced by of twice intraperitoneal injection excessive doses of L-arginine in mice / 中国药理学通报

AIM To establish a new, simple, stable and classical experimental model of acute edematous pancreatitis. METHODS Male mice were injected intraperitoneally 2?200 mg?(100 g) -1 body weight of L-arginine in an 1 h interval, as a 20% solution in 0.15 mol?L -1 NaCl. Control mice received the same quantity of 0.15 mol?L -1 NaCl. The mice were killed at 6, 12, 24, 48, 72 h following L-arginine administration. The serum amylase level, wet/dry weight ratio of the pancreas, histologic were assessed. RESULTS The serum level of amylase was significantly elevated at 6 h,reached the peak level at 24 h, normalized at 72 h. Histologic examination revealed interstitial edema and inflammatory cell infiltration reached the peak level at 24 h and decreased at 48,72 h. The wet/dry weight ratio of the pancreas changed in accordance with the interstitial edema. CONCLUSIONS The present study has demonstrated that the administration of excessive doses of arginine induces a new, noninvasive experimental model of acute edematous pancreatitis.