Formalin pre-fixation improves autopsy histology

Microscopic findings in key tissues are often critical to determine the cause of death in medical autopsies. The overall quality of histologic sections depends on numerous pre-analytic factors, among which are tissue section size and thickness. We designed a prospective quality improvement study to determine whether a simple intervention of formalin pre-fixation of myocardium, liver, and kidney tissues could improve the ease of cutting and quality of autopsy histologic sections as assessed by histotechnicians and pathologists. Of 46 autopsies included in the study, 21 were randomly assigned to formalin pre-fixation, and 25 underwent routine sectioning without formalin pre-fixation. A significant improvement in overall quality score by histotechnicians was detected in the sections from pre-fixed autopsy tissues compared to the control group (p=0.0327). There was no significant difference in quality score between the two groups as assessed by pathologists. Our autopsy quality improvement study demonstrates that a simple, low-cost intervention of formalin pre-fixation of fresh autopsy tissues for 90 minutes could significantly improve the overall quality of sections submitted for histologic processing.

Alcohol-based fixatives can better preserve tissue morphology than formalin / Fijadores a base de alcohol pueden preservar mejor la morfología tisular que la formalina

SUMMARY: Fixation is a crucial step in processing of tissue specimen for preservation of cellular architecture and composition of cells. Alcohol-based fixatives are considered some of the most promising alternatives to formalin. We evaluated the performance of alcohol-based fixatives (EthMeth and methacarn) and formalin as a comparator fixative in the research laboratory. Following 24 hours of fixation, tissue morphology and cellular details of the liver, spleen and brain (cerebral cortex) were evaluated. Morphological characteristics were evaluated by gross observations and analyzing cellular details, tissue architecture and overall staining characteristics (Hematoxylin and Eosin). EthMeth and methacarn fixation gave generally comparable and satisfactory results on the tissue morphology and subsequent identification of tissue characteristics. Particularly, tissues were well preserved and all nuclear as well as cytoplasmic details were clearly visible. However, formalin fixed tissues showed some peculiarity such as improper fixation, mild shrinkage, and alterations of tissue components. These results confirm that alcohol-based fixation is the superior alternative to formalin for preservation of tissue morphology. However, it is required to standardize the formalin-free methods and harmonize diagnosis in the laboratory worldwide.

RESUMEN: La fijación es un paso crucial en el procesamiento de muestras de tejido para preservar la arquitectura celular y la composición de las células. Los fijadores a base de alcohol se consideran algunas de las alternativas más prometedoras a la formalina. Evaluamos el rendimiento de los fijadores a base de alcohol (EthMeth y metacarn) y formalina como fijador comparativo en el laboratorio de investigación. Después de 24 horas de fijación, se observó la morfología del tejido y los detalles celulares del hígado, bazo y corteza cerebral. Se evaluaron las características morfológicas mediante observaciones generales y analizando detalles celulares, arquitectura de tejidos y características generales de tinción (hematoxilina y eosina). La fijación de EthMeth y metacarn dio resultados generalmente comparables y satisfactorios en la morfología del tejido y la posterior identificación de las características del mismo. Particularmente, los tejidos estaban bien conservados y todos los detalles nucleares y citoplasmáticos eran claramente visibles. Sin embargo, los tejidos fijados con formalina mostraron cierta peculiaridad, tal como una fijación inadecuada, la contracción leve y alteraciones de los componentes del tejido. Estos resultados confirman que la fijación a base de alcohol es la mejor alternativa a la formalina, para preservar la morfología del tejido. Sin embargo, es necesario estandarizar los métodos sin formalina y armonizar el diagnóstico en los laboratorios.

Brain structure morphology after being fixated with ethanol on electron microscope / Morfología de la estructura cerebral después de la fijación con etanol en microscopio electrónico

Fixation is one of the processes in preparing histology and pathology. The common material for fixation is buffered formalin including paraformaldehyde. However, the effect of the damaged cells, which is fixed for a long time, causes the research for other fixation materials to become necessary. In addition, paraformaldehyde is also harmful to human body and natural environment. Ethanol is one of the alternative fixation materials, which has been used for two hundred years. It has been used for many purposes, both in routine staining and immunohistochemistry. Nonetheless, no research confirms its effect on the electron microscope. The authors studied the effect of 50 % of ethanol on the cell membrane, organelles, and nucleus of Purkinje cells (Neuron purkinjense) observed on a light microscope and Transmitted Electron Microscope (TEM). Then it was compared to buffered formalin. In the light microscope, it shows that both of fixations have no different effects of the morphology of the cell membrane, cytoplasm, the nucleus of Purkinje cells and the neutrophils. We assume that our 50 % of ethanol concentration is almost the same as BF 10 % in the ability of hardening tissue and color absorption based on the previous study. In TEM, the structure of the cell membrane, organelles, and cytoplasm of Purkinje cell look broken in the cerebellum of 50 % of ethanol except for the nucleus. There was no significant difference diameter of the nucleus. It happened in general because of the shrinkage effect of ethanol. However, the authors recommend using 50 % of ethanol for routine staining.

La fijación es uno de los procesos en la preparación de muestras para histología y patología. El material más común para la fijación es la formalina tamponada. Sin embargo, el daño a las células que se mantienen en formalina durante mucho tiempo, hace necesario buscar otros materiales de fijación. Además, el paraformaldehido también es perjudicial para el cuerpo humano y el medio ambiente natural. El etanol es uno de los materiales de fijación alternativos que se ha utilizado durante muchos años, con diversos objetivos, tanto en la tinción de rutina como en la inmunohistoquímica. Sin embargo no se ha confirmdo su efecto con microscopio electrónico. Los autores estudiaron el efecto del 50 % de etanol sobre la membrana celular, los orgánulos y el núcleo de las células de Purkinje observados en un microscopio óptico y un microscopio de transmisión electrónico (TEM). Luego se comparó con la formalina tamponada. En el microscopio óptico se observó que ambas fijaciones no tienen efectos diferentes a la morfología de la membrana celular, el citoplasma, el núcleo de las células de Purkinje y los neutrófilos. Suponemos que nuestra concentración de 50 % de etanol es casi la misma que BF 10 % en la capacidad de endurecer el tejido y la absorción de color según el estudio anterior. En TEM, la estructura de la membrana celular, los orgánulos y el citoplasma de la célula de Purkinje presentaban daño en el cerebelo con un 50 % de etanol, a excepción del núcleo. No hubo diferencia significativa en el diámetro del núcleo. En general lo anterior se debió al efecto de contracción del etanol. En conclusión los autores recomiendan usar 50% de etanol para la tinción de rutina.

Conocimiento sobre la Citohistotecnología en una institución prestadora de servicios de salud IV / Knowledge on cytotechnology in a fourth level health care institution

Introducción: el Ministerio de Educación Nacional (MEN) de Colombia reconoció 420 instituciones de educación superior (IES), de las cuales 243 (57.8%) ofrecen programas técnicos o tecnológicos. El procesamiento de tejidos debe tener fundamento teórico y entrenamiento práctico para la preparación de tejidos humanos, animales o vegetales como apoyo diagnóstico. Esta profesión no es muy conocida y juega un papel relevante en el diagnóstico de patologías. Objetivo: determinar el conocimiento que tiene el personal de salud que labora en un hospital de cuarto nivel de atención con respecto a las habilidades y capacidades del citohistólogo. Métodos: estudio de corte transversal que incluyó trabajadores de una institución prestadora de servicios (IPS) de salud. Se calculó el tamaño muestral de 231 en Epi-info 7. Este trabajo fue aprobado por el comité de ética en investigación. Resultados: 231 trabajadores de la salud fueron interrogados, con una mediana de edad de 25 años (RIQ: 22-31), 83,1% mujeres, 61,5% conocen la definición correcta de citohistología y 49,3% lo relacionan con un aporte en promoción y prevención de cáncer. Conclusiones: existe un desconocimiento por parte de los trabajadores de la IPS sobre las actividades del citohistólogo y sus funciones, como la adecuada fijación, traslado y procesamiento de los tejidos, que al no realizarse en forma correcta afectan el diagnóstico patológico, lo cual incide en la morbilidad por muestras inadecuadas, causando además trastornos administrativos hospitalarios.

Introduction: the Colombian National Ministry of Education (MEN) recognized 420 higher education institutions (HEI) of which 243 (57.8%) offer technical and technological study programs. Human or animal tissue preparation and processing as aids to diagnosis requires understanding its theoretical principles and practical training. This profession is little known and plays a relevant role for diagnosing diseases. Objective: to determine the knowledge among health care personnel of a fourth level institution regarding the skills and competencies of a cytotechnologist. Methods: a cross-section study including workers of a health care institution (IPS). Epi-info 7 was used to calculate the sample size of 231. This work was approved by the research ethics committee. Results: 231 health care providers answered a questionnaire, median age 25 years (IQR: 22-31), 83.1% females, of which, 61.5% know the correct definition of cytotechnology and 49.3% relate cytotechnology with cancer prevention and control [screening] programs. Conclusions: the health care personnel of the institution were not acquainted with the activities and functions of a cytotechnologist or the tissue fixation, processing and transportation procedures. An inappropriate sample can affect pathology results impacting morbidity rates causing hospital administrative issues.

PLZFposc-KITpos-delineated A1-A4-differentiating spermatogonia by subset and stage detection upon Bouin fixation / 亚洲男科学杂志(英文版)

While hallmarks of rodent spermatogonia stem cell biomarkers' heterogeneity have recently been identified, their stage and subset distributions remain unclear. Furthermore, it is currently difficult to accurately identify subset-specific SSC marker distributions due to the poor nuclear morphological characteristics associated with fixation in 4% paraformaldehyde. In the present study, testicular cross-sections and whole-mount samples were Bouin fixed to optimize nuclear resolution and visualized by immunohistochemistry (IHC) and immunofluorescence (IF). The results identified an expression pattern of PLZFhighc-KITpos in A1 spermatogonia, while A2-A4-differentiating spermatogonia were PLZFlowc-KITpos. Additionally, this procedure was used to examine asymmetrically expressing GFRA1 and PLZF clones, asymmetric Apr and false clones were distinguished based on the presence or absence of TEX14, a molecular maker of intercellular bridges, despite having identical nuclear morphology and intercellular distances that were <25 μm. In conclusion, this optimized Bouin fixation procedure facilitates the accurate identification of spermatogonium subsets based on their molecular profiles and is capable of distinguishing asymmetric and false clones. Therefore, the findings presented herein will facilitate further morphological and functional analysis studies and provide further insight into spermatogonium subtypes.

Tratamento das ptoses mamárias com implantes submusculares e pontos de fixação do tecido mamário ao muscular: aspectos técnicos e avaliação de resultado / Treatment of breast ptosis with submuscular implants and points of attachment of the breast to the pectoralis muscle: technical considerations and outcome assessment

Introdução: Mastopexia associada à inclusão de implante é uma situação desafiadora para o cirurgião plástico. O objetivo é descrever a colocação de implante submuscular com descolamento anatômico mais pexia firme do tecido glandular usando pontos de fixação do tecido mamário ao muscular e analisar os resultados estéticos das pacientes operadas. Método: Foram realizadas 23 mastopexias com implantes no período entre abril de 2015 e julho de 2017, pelo mesmo cirurgião, sendo as mamas das pacientes marcadas previamente, na posição sentada. Realizou-se incisão no sulco mamário e descolamento até o polo superior da mama no plano subfascial, fixação da glândula ao músculo peitoral maior com 9 a 12 pontos. A seguir, iniciou-se a dissecção do músculo peitoral maior através de sua origem costal e transição com os músculos reto abdominal e serrátil, liberando amplamente na porção inferior. Introduziu-se o implante e completou-se a mastopexia. Os tamanhos dos implantes variaram de 255ml a 355ml. Fotos das mamas de 12 pacientes foram avaliadas por dois cirurgiões plásticos e dois leigos, nos seguintes parâmetros: resultado estético, simetria das aréolas e grau de ptose mamária. As avaliações podiam ser Ruim, Razoável ou Bom. Resultados: A técnica cirúrgica mostrou-se reprodutível, apenas 1 caso de hematoma unilateral, nenhuma infecção, queixas de dor discretas. Apenas um caso foi considerado, por um único avaliador, como Razoável; as demais avaliações consideradas como Bom. Conclusão: O tratamento de ptoses mamárias com colocação de implante submuscular acrescido de pexia da glândula ao músculo peitoral é uma técnica reprodutível e com bons resultados estéticos.

Introduction: Mastopexy associated with implant placement is challenging for plastic surgeons. The objective is to describe the placement of a submuscular implant with anatomical detachment in combination with stable fixation of the breast tissue to the pectoralis muscle and analyze the aesthetic results. Method: Twenty-three mastopexy procedures with implants were performed from April 2015 to July 2017 by the same surgeon, and surgical markings were made in the breasts of the patients in a seated position. An incision was made in the inframammary fold, and the breast tissue was elevated to the upper pole in the subfascial plane and attached to the pectoralis major muscle using 9-12 stitches. Subsequently, the inferior margin of the pectoralis major muscle and the transition from the rectus abdominis muscle to the serratus muscle were dissected to expose the muscle. The implant was introduced and mastopexy was completed. Implant size ranged from 255 mL to 355 mL. Photographs of the breasts of 12 patients were evaluated by two plastic surgeons and two non-medical subjects, who considered the aesthetic results, symmetry of the nipple-areola complex, and degree of breast ptosis. The results were scored as unsatisfactory, satisfactory, or good. Results: The surgical technique was reproducible; there was only one case of unilateral hematoma, no implant infections, and only complaints of mild pain. Only one case was scored as satisfactory by one evaluator, whereas the results of the other cases were considered good. Conclusion: The treatment of breast ptosis with the placement of a submuscular implant in combination with fixation of the breast to the pectoralis major muscle is reproducible and yields good aesthetic results.

A comparative study of fixatives for axolotl blastema tissue / Estudio comparativo de fijadores para el tejido blastema del axolotl

Regeneration is defined as tissue renewal and functional restoration process of the damaged parts of the body after an injury. Ambystoma mexicanum, commonly named the Axolotl, is one of the unique vertebrates, which has a remarkable ability to regenerate their extremities following the amputation. Although the process of regeneration includes several periods, it can be divided into two main phases; blastema formation and dedifferentiation. In the couple of hours following the amputation, wound closure occurs by migration of epithelial cells around the amputation site followed by macrophage infiltration and dedifferentiation of cells to turn into stem cells. Accumulated stem cells form a very authentic tissue type called blastema, which is crucial for successful regeneration. In order to evaluate this exceptional tissue and acquire high quality images, it is crucial to employ specific procedures to prepare the tissue for imaging. Here, in this study, we aimed to investigate success of various fixative solutions (Carnoy's, Bouin's, % 10 NBF, Clarke's, Alcoholic Formaline and AFA) to monitor the fixed blastema. Our data reveals that integrity of the blastema tissue differs among used fixatives and a significant difference is observed between the samples in terms of staining quality.

La regeneración se define como la renovación del tejido y el proceso de restauración funcional de las partes dañadas del cuerpo después de una lesión. Ambystoma mexicanum, comúnmente llamado Axolotl, es uno de los únicos vertebrados que tiene una notable capacidad para regenerar sus miembros después de una amputación. Aunque el proceso de regeneración incluye varios períodos, se puede dividir en dos fases principales: formación del blastema y desdiferenciación. En el par de horas después de la amputación, el cierre de la herida ocurre por la migración de células epiteliales alrededor del sitio de la amputación seguido por una infiltración de macrófagos y la desdiferenciación de las células para convertirse en células madre. Las células madre acumuladas forman un tipo de tejido muy diferenciado denominado blastema, que es crucial para una exitosa regeneración. Para evaluar este tejido y adquirir imágenes de alta calidad, es crucial emplear procedimientos específicos para la obtención de imágenes. En este estudio, se intentó investigar el éxito de varias soluciones fijadoras (Carnoy, Bouin, % 10 NBF, Clarke, Formalina Alcohólica y AFA) para monitorear la fijación del blastema. Nuestros datos revelan que la integridad del tejido del blastema difiere entre los fijadores utilizados y una diferencia significativa observada entre las muestras se da en términos de la calidad de tinción.

Optimizing Fixation Protocols to Improve Molecular Analysis from FFPE Tissues

Abstract Most Departments of Pathology around the world have a considerable archive of formalin-fixed paraffin-embedded (FFPE) tissue suitable for molecular assessment. This article points out the potential DNA damage that may occur if basic steps are not followed during processing and storage of these samples. Furthermore, it hopes to establish parameters to optimize quality and quantity of DNA extracted from FFPE tissues.

Resumo A maioria dos Departamentos de Patologia em todo o mundo têm um considerável acervo de tecidos embebidos em parafina e fixados em formalina, que são passíveis para análises moleculares. Este artigo apresenta os danos ao DNA que podem ocorrer se passos básicos não forem seguidos durante o processamento e armazenamento destas amostras. Além disso, procura estabelecer parâmetros para otimizar a qualidade e quantidade do DNA extraído de tecidos FFPE.

Fita microporosa para fixação de enxertos cutâneos / The use of a microporous tape for fixing skin grafts

Introdução: A fixação do enxerto cutâneo é essencial para sua integração no leito receptor. A literatura apresenta várias técnicas de fixação, porém, o uso da fita de microporosa é pouco relatado. O objetivo é demonstrar e divulgar o uso da fita microporosa na fixação do enxerto cutâneo Métodos: Estudo prospectivo, realizado de janeiro de 2014 a janeiro de 2016. Em 40 pacientes foi utilizada a fita microporosa esterilizada como método isolado para a fixação do enxerto Resultados: Enxertos cutâneos apresentaram resultado satisfatório sem mobilização e, consequentemente, boa integração. Conclusão: O uso da fita microporosa esterilizada é um excelente método para a fixação de enxertos cutâneos, por ser simples, rápido e seguro.

Introduction: Fixing a skin graft is essential to its integration in the recipient bed. The literature presents several fixation techniques. However, only few reports on the use of microporous tape are available. This study aims to demonstrate and promote the use of microporous tape in fixing skin grafts. Methods: A prospective study was performed from January 2014 to January 2016. In 40 patients, a sterilized microporous tape was used as an isolated method to fix skin grafts. Results: The use of skin graft immobilization showed satisfactory results and consequently good integration. Conclusion: The use of a sterilized microporous tape is an excellent method for fixing skin grafts because it is easy, fast, and safe to use.

Eficacia del diagnóstico diferencial de tuberculosis mediante reacción de polimerasa en cadena en lesiones granulomatosas de tejidos fijados en formalina e incluidos en parafina / Efficacy of PCR for the differential diagnosis of tuberculosis in granulomatous lesions of paraffin-embedded formalin fixed tissues

Background: Granulomatous lesions occur in tuberculosis (TB), other infections, toxic, allergic, and autoimmune diseases among others. In absence of a an acid-fast bacilli (AFB) confirmation of TB is necessary. Objective: To assess the efficacy of PCR for TB detection and to correlate with granuloma histology and AFB staining. Methods: We analyzed 380 fixed paraffin-embedded tissues (PETs) of granulomas with and without caseous necrosis; suppurative; sarcoidal; or of chronic nonspecific nature. Nested PCR-IS6110 for Mycobacterium tuberculosis complex (MTB) and a nested pan-Mycobacterium for the hsp65 gene were used for Mycobacterium spp detection. Results: PCR was more sensitive than AFB staining for all five catageories of granulomas: G1: PCR 71%, AFB staining 28%. G2: PCR 37%, AFB 8%. G3: PCR 17%, AFB staining 7%. G4: PCR 8%, AFB staining 4%. G5: PCR 6%, AFB staining 0%. Conclusions: Molecular diagnosis of TB using PCR-based testing is a fast, efficacious and sensitive method that increased the accuracy of PET histological diagnosis associated with granulomatous lesions.

Introducción: Lesiones granulomatosas ocurren en tuberculosis (TBC), otras infecciones, condiciones tóxicas, alérgicas y autoinmunes, entre otras. Con baciloscopia negativa, es necesario confirmar el diagnóstico de TBC. Objetivo: Evaluar la eficacia de la RPC para detectar TBC comparado con baciloscopia en relación a la histología del granuloma. Métodos: Analisis de 380 tejidos fijados en formalina e incluidos en parafina (TFFP) con diferentes tipos de granulomas: con necrosis caseosa; sin necrosis caseosa; supurativo; sarcoidal; a cuerpo extraño/inespecífico. Utilizamos RPC anidada-IS6110 para detección del complejo Mycobacterium tuberculosis (MTB) y una pan-RPC anidada-hsp65 para Mycobacterium spp. Resultados: La detección de TBC mediante RPC fue significativamente superior a baciloscopia en los cinco tipos de granuloma: G1: RPC 71%, baciloscopia 28%; G2: RPC 37%, baciloscopia 8%; G3: RPC 17%, baciloscopia 7%; G4: RPC 8%, baciloscopia 4%; G5: RPC 6%, baciloscopia 0%. Conclusión: El diagnóstico de TBC por RPC es un método rápido, eficaz y de gran sensibilidad, que aumenta la precisión del diagnóstico diferencial de lesiones granulomatosas de TFFP procesados rutinariamente en histopatología.

Difference between actual vs. pathology prostate weight in TURP and radical robotic-assisted prostatectomy specimen

Introduction To investigate and highlight the effect of formaldehyde induced weight reduction in transurethral resection of prostate (TURP) and radical robotically-assisted prostatectomy (RALP) specimen as a result of standard chemical fixation. Materials and Methods 51 patients were recruited from January 2013 to June 2013 who either underwent a TURP (n=26) or RALP (n=25). Data was collected prospectively by the operating surgeon who measured the native, unfixed histology specimen directly after operation. The specimens were fixed in 10% Formaldehyde Solution BP and sent to the pathology laboratory where after sufficient fixation period was re-weighed. Results Overall mean age 64.78 years, TURP mean age 68.31 years RALP mean age 61.12years. We found that the overall prostatic specimen (n=51) weight loss after fixation was a mean of 11.20% (3.78 grams) (p≤0.0001). Subgroup analysis of the native TURP chips mean weight was 16.15 grams and formalin treated mean weight was 14.00 grams (p≤0.0001). Therefore, TURP chips had a mean of 13.32 % (2.15 grams) weight loss during chemical fixation. RALP subgroup unfixed specimen mean weight was 52.08 grams and formalin treated mean weight was 42.60 grams (p≤0.0001), a 19.32 % (9.48grams) mean weight reduction. Conclusion It has not been known that prostatic chips and whole human radical prostatectomy specimen undergo a significant weight reduction. The practical significance of the accurate prostate weight in patient management may be limited, however, it is agreed that this should be recorded correctly, as data is potential interest for research purposes and vital for precise documentation. .

HIGH SENSITIVE PCR METHOD FOR DETECTION OF PATHOGENIC Leptospira spp. IN PARAFFIN-EMBEDDED TISSUES / PCR altamente sensible para la detección de Leptospira spp. patógenas en tejidos embebidos en parafina

This study describes the development and application of a new PCR assay for the specific detection of pathogenic leptospires and its comparison with a previously reported PCR protocol. New primers were designed for PCR optimization and evaluation in artificially-infected paraffin-embedded tissues. PCR was then applied to post-mortem, paraffin-embedded samples, followed by amplicon sequencing. The PCR was more efficient than the reported protocol, allowing the amplification of expected DNA fragment from the artificially infected samples and from 44% of the post-mortem samples. The sequences of PCR amplicons from different patients showed >99% homology with pathogenic leptospires DNA sequences. The applicability of a highly sensitive and specific tool to screen histological specimens for the detection of pathogenic Leptospira spp. would facilitate a better assessment of the prevalence and epidemiology of leptospirosis, which constitutes a health problem in many countries.

El presente estudio describe el desarrollo y aplicación de un nuevo ensayo de PCR para la detección específica de leptospiras patógenas y su comparación con un protocolo reportado previamente. Se diseñaron nuevos cebadores para la optimización y evaluación de la PCR en tejidos embebidos en parafina infectados artificialmente. La PCR se aplicó además a muestras de tejidos embebidos en parafina y se realizó la secuenciación del amplicón resultante. La PCR diseñada fue más eficiente que el protocolo reportado, permitiendo la amplificación del fragmento de ADN esperado en las muestras infectadas artificialmente y del 44% de las muestras post mortem. Se secuenciaron 10 amplicones provenientes de pacientes diferentes. La aplicabilidad de una herramienta altamente sensible y específica en la búsqueda de leptospiras patógenas en especímenes histopatológicos podría facilitar una mejor valoración de la prevalencia y la epidemiología de la leptospirosis, la que constituye un problema de salud en disímiles países.

Epidermal growth factor receptor gene expression evaluation in colorectal cancer patients.

Background: Colorectal cancer is one of the most common causes of death in the world and third and fourth most common cancer among men and women in Iran respectively. Epidermal growth factor receptor (EGFR) is a tyrosine kinase receptor that shows over expression in epithelial tumors and regulates important processes in tumorigenesis. Incidence and characteristics of colorectal cancer are based on the geographic region and race. Aim: In this research work, the over expression of EGFR in formalin fixed paraffin-embedded (FFPE) colorectal cancer tumor tissue of patients was studied. Materials and Methods: Fifteen FFPE colorectal cancer tumor tissues (10 women and 5 men; 25-65 years old and stage IV) and 15 non-patients (nine women and six men; 25-65 years old) that were collected during 2006-2012. EGFR gene expression level was analyzed by real-time quantitative reverse transcriptase polymerase chain reaction (PCR). All PCR reactions were performed in triplicate for both target gene and internal control (18s ribosomal ribonucleic acid) with the 2−ΔΔCT method. Gene expression differences in patients and controls were evaluated with t-test. Results: The results were showed EGFR gene over expression in 12 (80%) of 15 patients. There was a statistically significant difference in the prevalence of EGFR expression between patients and control (P < 0.05). Conclusion: Our results demonstrated EGFR gene over expression in colorectal cancer tumor tissue compared with controls.

Origem e principais ramificações da artéria celíaca em avestruz (Struthio camelus, Linnaeus, 1758) / Origin and main ramifications of the celiac artery of the ostrich(Struthio camelus, Linnaeus, 1758)

Foram estudados os arranjos da artéria celíaca em 30 filhotes de avestruzes (15 machos e 15 fêmeas). O comprimento médio da artéria celíaca foi 0,33±0,08cm nos machos e 0,32±0,14cm nas fêmeas, não havendo diferença nesta medida entre sexos. Não houve correlação entre o comprimento rostrossacral e o comprimento da artéria celíaca em ambos os sexos. Sua origem ocorreu ao nível do sétimo espaço intercostal na maioria dos casos, ainda que a esqueletopia tenha variado independentemente do sexo.Seu território de irrigação incluiu o esôfago, pró-ventrículo, ventrículo, baço, fígado, pâncreas, duodeno, jejuno, íleo e cecos. A artéria celíaca origina-se da aorta descendente e fornece as artérias pró-ventricular dorsal e esplênica para posteriormente se dividir em ramos esquerdo e direito. Na maioria dos avestruzes, o ramo esquerdo ofereceu ramos para o esôfago, pró-ventrículo e ventrículo em padrões variados. O ramo direito irrigou inicialmente o pâncreas, emitiu uma artéria hepática direita para o fígado,uma artéria gástrica direita para o ventrículo e terminou como artéria pancreatico duodenal para o pâncreas e porções do duodeno.Finalmente, esta artéria emitiu numerosos ramos ileocecais para o íleo e cecos direito e esquerdo. Artérias duodenojejunais e jejunais surgiram apenas em 10% e 3,33% dos animais, respectivamente. Todos apresentaram a artéria marginalis intestini tenuispercorrendo a margem mesentérica do intestino delgado. Anastomoses de ramos da artéria celíaca com os oriundos da mesentérica cranial ocorreram em 20% dos casos.

The arrangement formed by the celiac artery in 30 ostrich chicks, 15 males and 15 females were studied. The average length of the celiac artery was 0.33±0.08 cm for male and 0.32±0.14cm in females, even though there was no difference of this measurement between genders. There was also no correlation between rostrossacral length and the length of the celiac artery in both sexes. Its origin was at the level of seventh intercostal space in most cases, despite a varied skeletopy which was independent of gender. Its territory of irrigation included esophagus, proventriculus, gizzard, liver, spleen, pancreas, duodenum, jejunum, ileum and bothcecum. The celiac artery leaves the aorta and originates pro-ventricular dorsal artery and splenic artery and thus splits into two other branches: left and right. In most ostriches, left branch offered branches to the esophagus, proventriculus and gizzard in severaldifferent patterns. The right branch irrigated first the pancreas and formed a right hepatic artery to the liver, a right gastric artery to the gizzard and finished as pancreatic duodenalis artery to supply the pancreas and the ascendens and descendens portions from duodenum. Finally, this artery has formed numerous ileocecal branches to ileum and to right and left cecum. Duodenojejunal and jejunal arteries were present only in 10% and 3.33% of the ostriches, respectively. All animals showed intestini tenuis marginalisartery transiting the mesenteric border of the small intestine. Anastomoses of the intestinal celiac branches with those from cranial mesenteric artery were found in 20% of cases.

Delay in formalin fixation and HER2 testing in gastric cancer / 中华病理学杂志

<p><b>OBJECTIVE</b>To evaluated HER2 status using immunohistochemistry (IHC) assay and fluorescence in situ hybridization (FISH) at two different time points of tissue fixation after surgical resection of gastric cancer, emphasizing the importance of standard operation and quality control in HER2 testing.</p><p><b>METHODS</b>Forty-one resection specimens of advanced gastric cancer were collected with tissue fixation periods of < 30 min or > 30 min after surgical resection. HER2 status was evaluated by immunohistochemistry (IHC) assay and fluorescence in situ hybridization (FISH).</p><p><b>RESULTS</b>The frequency of HER2 expression by IHC in the samples with fixation time of < 30 min was higher than that in those of > 30 min (P < 0.05). However, no significant difference was observed by FISH (P > 0.05) between the two groups. Samples of < 30 min fixation time had high concordant results between IHC and FISH (100.0% for both positive and negative cases, Rho = 0.724, P < 0.05). In addition, HER2 expression by IHC was significantly correlated with Lauren classification, histologic differentiation, TNM stage and gender (P < 0.05).</p><p><b>CONCLUSION</b>The time to tissue fixation after surgical resection of more than 30 min has deleterious effect on the detection of HER2 by IHC although FISH testing is not affected.</p>

Arthroscopic Biceps Tenodesis: Soft Tissue versus Bone Fixation / 대한스포츠의학회지

The purpose of our study was to compare the clinical results between arthroscopic bone fixation on intertubercular groove using suture anchor and soft tissue fixation at the rotaor interval for biceps tenodesis when partial tear or instability of biceps tendon accompanied with rotator cuff tear. From January 2010 to January 2012, 34 cases who were performed biceps tenodesis for partial tear or instability were enrolled in our study. Mean follow-up period was 30.2 months. Bone fixation using suture anchor was performed in 18 cases, and soft tissue fixation was performed in 16 cases. Clinical result was evaluated by pain visual analogue scale (VAS), Speed test, Yergason test, muscle strength, and Constant score. Pain VAS of cases with soft tissue fixation was significantly higher than that of cases with bone fixation at 6 months and final follow-up. Positive results for the final follow-up Speed and Yergason test were checked in 4 cases (25%) with soft tissue fixation and 1 (5.6%) with bone fixation. The Popeye deformity was seen in 4 cases (25%) with soft tissue fixation and 2 (11%) with bone fixation. Constant score was improved 47 to 78 in cases with soft tissue fixation and 48 to 86 in cases with bone fixation. In patient with partial tear or instability of biceps tendon accompanied with rotator cuff tear, biceps tenodesis using soft tissue fixation showed worse result compared with bone fixation because of long duration of the pain. Therefore, when performing the biceps tenodesis, bone fixation will be recommended.

Fibrin Glue Fixation for Suction Blister Epidermal Grafting in Two Patients with Stable Vitiligo

Vitiligo is a chronic disorder characterized by depigmented macules which can slowly enlarge with the concurrent development of new lesions. Although autologous suction blister epidermal grafting is an established technique for the treatment of recalcitrant, stable vitiligo, the donor tissue graft is not easy to fix at the recipient site, especially in areas such as the joints, face, cutaneous folds, hands, feet, and hair-bearing areas. Therefore, various methods of donor tissue fixation have been attempted. We report two cases of vitiligo treated with suction blister epidermal grafting, with fibrin tissue adhesion. The first case is that of 16-year-old female patient presented with hypopigmented patches on the forehead and frontal scalp area. The other case is that of 32-year-old female patient presented with hypopigmented patches on the chin. We treated them with phototherapy for 1~4 years; however, the lesions were recalcitrant. Therefore, we tried treatment with a suction blister epidermal graft. Because graft fixation is difficult at the recipient sites, fibrin glue was sprayed on the grafts. Thereafter, we applied a porous silicone wound contact layer over the graft area and applied sterile gauze dressing that was left for a week. One week after the procedure, firm fixation of the donor tissue was observed in both cases. Fibrin glue seemed to improve the graft fixation, providng protection against infection and an optimal environment for wound healing. This report suggests that the application of an epidermal graft with fibrin glufixation, can provide the best result in the surgical treatment of stable vitiligo.

Laser microdissection and mass spectrometry based proteomics in the diagnosis of kidney diseases / 生物工程学报

In recent years, laser microdissection followed by mass spectrometry (LMD/MS) has been successfully applied to the proteomic studies of formalin-fixed paraffin-embedded (FFPE) renal tissues. This new technique improves the diagnosis of kidney diseases and has a better potential for future clinical application. The review focuses on the use of this methodology for exploring the mechanisms, diagnosis and classification of kidney diseases including renal amyloidosis and membrane proliferative glomerulonephritis.

Experimental model for the study of soft tissue fixation methods on skin-pericranium flaps in rats

PURPOSE: To develop an experimental model to study and radiologically monitor displacement of skin flaps in the pericranium of rats subjected to traction and surgical fixation using suture anchored in a skull bone tunnel or with N-butyl-2-cyanoacrylate (HistoacrylTM) surgical adhesive. METHODS: Radiological markers were placed in the subcutis of Wistar rats undergoing subperiosteal detachment of the pericranium with pulling and fixation of the flap. We performed radiography on postoperative days 3, 7, 14, 21, and 45. A p-value of <0.05 was considered significant. RESULTS: Qualitative analysis of the data indicated that the flaps in the surgical adhesive group remained in place with no change from the immediate postoperative position. However, the flaps in the suture anchored in the skull bone tunnel group and in the control group showed similar healing, with a loss of attachment of 9.7% and 22%, respectively, compared with the immediate postoperative position. There was no quantitative difference between the groups. CONCLUSIONS: This experimental model created acceptable experimental conditions for testing different soft tissue fixation methods. The use of tissue fixatives contributed to better placement of pericranium-cutaneous flaps, and surgical adhesive was superior to suture anchor in the skull bone tunnel for fixation of pericranium-cutaneous flaps.

Correção de flacidez de pele nas regiões glútea e lombar com retalho fasciocutâneo local em cambalhota / Correction of sagging skin the gluteal and lumbar regions using a fasciocutaneous local flap

Um dos maiores problemas dos pacientes pós-cirurgia bariátrica é o excedente de pele nas regiões glútea e lombar. Somado a isso, há escassez de tecido para preencher a região glútea, que pode ser corrigida por vários métodos, entre eles a lipoenxertia ou, ainda, o implante de prótese isoladamente, nos casos em que não haja ptose. Se o contrário ocorre, essas opções tornam-se muito limitadas e pouco expressivas. O lifting glúteo com emprego de retalho fasciocutâneo em cambalhota permite resolver tanto a flacidez como o excesso de pele com ptose. Este trabalho tem por objetivo demonstrar a facilidade de confecção e a versatilidade de utilização do retalho glúteo em cambalhota para correção de flacidez das regiões glútea e lombar. Uma paciente foi submetida ao procedimento cirúrgico de lifting glúteo associado ao emprego de retalho fasciocutâneo desepidermizado em cambalhota, para correção de falta de preenchimento local. Melhora acentuada das regiões glútea e lombar foi obtida após a retirada do excesso de tecido superior, confecção de um retalho fasciocutâneo de pedículo inferior e tração da pele. Além disso, a rotação do retalho e sua fixação ao músculo glúteo melhoraram acentuadamente o contorno posterior da paciente, com pós-operatório bastante cômodo. Com morbidade mínima, foi retirado o excesso cutâneo e, concomitantemente, aumentada a região glútea com retalho autólogo, que é desprezado durante a execução de muitas técnicas, melhorando o contorno da paciente e deixando uma cicatriz discreta, que poderá ser ocultada pelas roupas íntimas.

A major problem for patients after bariatric surgery is the excess skin in the gluteal and lumbar regions. In addition, there is a lack of available tissue to fill the gluteal region, which can be overcome by various methods, including fat grafting or even prosthesis implantation alone, in cases where no ptosis is noted. However, if this is not the case, the options become very limited and those available are not very effective. The use of a buttock lift with a fasciocutaneous flap solves the problems of sagging and excess skin with ptosis. In the present article, we aimed to demonstrate the ease of preparation and the versatility of use of the gluteal flap to correct sagging of the lumbar and gluteal regions. One patient underwent a buttock lift using a de-epidermized fasciocutaneous flap to overcome the lack of local filling. Marked improvement of the gluteal and lumbar regions was noted after removal of the excess tissue, construction of an inferior fasciocutaneous flap, and use of skin traction. In addition, rotation of the flap and its attachment to the gluteal muscle markedly improved the patient's posterior contour and postoperative comfort. With minimal morbidity, excess skin was removed and the gluteal region was concomitantly increased using an autologous flap that is neglected in many techniques; this resulted in an improved contour and a discreet scar that could be hidden by the patient's underwear.