Micro-plastination: technique for obtaining slices below 250 µm for the visualization of microanatomy in morphological and pathological morphology protocols / Micro-plastinación: técnica para la obtención de cortes inferiores a 250 µm para la visualización de la microanatomía en protocolos experimentales de morfología y patología

Plastination has revolutionized the study and research of anatomy, thanks to the biosecurity and indefinite preservation of human and animal bodies and organs. This paper presents the concept of Micro-Plastination, an ultra-thin sheet plastination technique, to obtain ultra-thin slices, of a thickness of less than 250 µm, for the identification and visualization of the microanatomy of any anatomical region in morphological and pathological experimental protocols.

La plastinación ha revolucionado el estudio y la investigación de la anatomía, gracias a la conservación biosegura y por tiempo indefinido de cadáveres y órganos humanos y animales. En este trabajo se presenta el concepto de Micro-Plastinación, técnica de plastinación de cortes ultrafinos para la obtención de cortes ultradelgados, de un grosor inferior a los 250 µm, para la identificación y visualización de la microanatomía de cualquier región anatómica en protocolos de morfología experimental.

Repair of Inaccessible Ventral Dural Defect in Thoracic Spine: Double Layered Duraplasty

We propose a double layered (intradural and epidural patch) duraplasty that utilizes Lyoplant and Duraseal. We examined a 47-year-old woman after decompression for thoracic ossification of posterior longitudinal ligament was performed in another hospital. On postoperative day 7, she complained of weakness in both legs. Postoperative magnetic resonance imaging (MRI) showed cerebrospinal fluid (CSF) collection with cord compression. In the operative field, we found 2 large dural defects on the ventral dura mater. We performed a conventional fat graft with fibrin glue. However, the patient exhibited neurologic deterioration, and a postoperative MRI again showed CSF collection. We performed dorsal midline durotomy and inserted a intradural and epidural Lyoplant patch. She immediately experienced diminishing back pain postoperatively. Her visual analog scale and motor power improved markedly. Postoperative MRIs performed at 2 and 16 months showed no spinal cord compression or CSF leakage to the epidural space. We describe a new technique for double layered duraplasty. Although we do not recommend this technique for all dural repairs, double-layered duraplasty may be useful for repairing large inaccessible dural tears in cases of persistent CSF leakage refractory to conventional management.

Downregulation of inwardly rectifying potassium channel 5.1 expression in C57BL/6J cochlear lateral wall / 华中科技大学学报（医学）（英德文版）

Age-related hearing loss (AHL) is one of the most common sensory disorders among elderly persons. The inwardly rectifying potassium channel 5.1 (Kir5.1) plays a vital role in regulating cochlear K(+) circulation which is necessary for normal hearing. The distribution of Kir5.1 in C57BL/6J mice cochleae, and the relationship between the expression of Kir5.1 and the etiology of AHL were investigated. Forty C57BL/6J mice were randomly divided into four groups at 4, 12, 24 and 52 weeks of age respectively. The location of Kir5.1 was detected by immunofluorescence technique. The mRNA and protein expression of Kir5.1 was evaluated in mice cochleae using real-time polymerase-chain reactions (RT-PCR) and Western blotting respectively. Kir5.1 was detected in the type II and IV fibrocytes of the spiral ligament in the cochlear lateral wall of C57BL/6J mice. The expression levels of Kir5.1 mRNA and protein in the cochleae of aging C57BL/6J mice were down-regulated. It was suggested that the age-related decreased expression of Kir5.1 in the lateral wall of C57BL/6J mice was associated with hearing loss. Our results indicated that Kir5.1 may play an important role in the pathogenesis of AHL.

Avaliação in vitro, com microscopia óptica, da incidência de istmos em molares inferiores / In vitro evaluation with optical microscopy of isthmus incidence in mandibular molars

A proposição desse estudo foi verificar a incidência da fusão dos canais, tanto nas raízes mesiais quanto nas distais, em molares inferiores. Esse estudo foi aprovado pelo Comitê de Ética da Universidade de Fortaleza sob o número 181.388, de 20 de dezembro de 2012. Para a pesquisa foram selecionados 150 molares inferiores, sendo 50 primeiros molares (Grupo I), 50 segundos molares (Grupo II) e 50 terceiros molares (Grupo III). As raízes de todos os dentes foram seccionadas com disco de carborundum. Para melhor visualização da área de istmo, foi utilizada magnificação visual com microscopia óptica e limas manuais. Com a metodologia empregada, verificou-se que: no Grupo I, 76% dos canais mesiais e 53,7% dos distais possuíam comunicação; no Grupo II, 86% dos canais mesiais e 40,6% dos distais apresentaram área de istmo entre os canais radiculares; por fim, no Grupo III, 91% dos canais mesiais e 13,5% dos distais se comunicavam por istmos. Assim, por meio da metodologia empregada, foi possível verificar que a incidência de istmos conectando os canais radiculares presentes nas raízes dos molares inferiores foi expressiva nos três grupos.

Melanocortin-4 receptor expression in the cuneiform nucleus is involved in modulation of opioidergic signaling / 华中科技大学学报（医学）（英德文版）

Substantial evidence has suggested that deep brain stimulation of the cuneiform nucleus has become a remarkable treatment option for intractable pain, but the possible mechanism is poorly understood. Using a melanocortin-4 receptor (MC4R)-green fluorescent protein (GFP) reporter knockin mouse, we showed that a large number of MC4R-GFP-positive neurons were expressed in the cuneiform nucleus. Immunofluorescence revealed that approximately 40%-50% of MC4R-GFP-positive neurons expressed mu opioid receptors, indicating that they were opioidergic signaling. Our findings support the hypothesis that MC4R expression in the cuneiform nucleus is involved in the modulation of opioidergic signaling.

Immunohistochemical detection of axillary lymph node micrometastases in breast cancer patients: Increasing the accuracy of detection and decreasing labor intensive serial sectioning.

Background: The histological detection of axillary lymph node tumor metastases in cases of breast carcinoma is of major prognostic significance, but may be difficult when metastases are of microscopic size. The micrometastases can be detected either by immunohistochemistry (IHC) or serial sectioning. Aims: We investigated whether immunohistochemical techniques and serial sectioning can increase the accuracy of metastatic detection and compared the efficacy of both. Materials and Methods: Thirty cases of breast carcinoma were studied in all of whom the axillary lymph nodes had been reported as free of metastases. Blocks from these cases were serially sectioned and stained with hematoxylin and eosin and a single section was stained with monoclonal antibody to cytokeratin AE1/AE3 and epithelial membrane antigen. The positivity for micrometastases was correlated with size, number, grade and histological type of primary tumor, lymph node size and number. Results and Conclusion: In 5/30 previously unsuspected cases, micrometastases were revealed by IHC and in 1/30 by serial sectioning. These findings suggested that serial sectioning is a labor intensive, time consuming and impractical procedure. Micrometastases were more frequently detected with age of patient >50 years, Grade 2/3 tumor, tumor size >5 cm and more than one primary tumor. Immunohistochemical analysis can be recommended as a routine procedure or an adjunct to routine histological procedures for the correct staging of breast carcinoma and use of adjuvant chemotherapy, especially in the high risk group.

Establishment of localization ultrathin section for cytopathic cells / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish a localization ultrathin section method through which target cytopathic cells could be sectioned in situ.</p><p><b>METHODS</b>Lab-Tek Chamber slide system (177402) was selected as resin embedding mould. Cells infected with Human adenovirus type 5 (Ad5) or A/HN/SWL3/ 2009 (H1N1) influenza virus were embedded in situ as models. Target cytopathic cells were exposed by trimming, sectioned and observed under transmission electron microscope (TEM).</p><p><b>RESULTS</b>Target cells could be sectioned in situ and virus particles could be found easily on sections.</p><p><b>CONCLUSION</b>A localization ultrathin sectioning method was established and this technique could be applied in virus detection in cytopathic cells to improve TEM detection efficiency.</p>

Role of dilute alcohol in the removal of fine wrinkles from paraffin sections, a histo-technical study / Rol del alcohol diluido en la eliminación de arrugas finas desde secciones de parafina, un estudio histotécnico

Histotechnology is concerned with processing and preparing of the body tissue in such a manner as to enable a satisfactory study of it. Section cutting is an integral part of histology and histopathology. It is an art by itself requiring skill and knowledge on the part of technician or the person who needs to do section cutting. In the routine method of preparing paraffin sections, it is often encountered by the presence of artefacts like fine wrinkles or folds. Attempts have been made to remove the wrinkles by floating the sections in the warm water bath. However this method has not been able to remove all the wrinkles from the sections. We have designed a simple and reliable method, in which the paraffin sections were floated over the ethyl alcohol diluted with water (1:15) before they were placed in the water bath. Through this method, we have been able to get the wrinkle free sections of superior quality. The advantage of our method is that, it is easy to prepare the dilute alcohol and is cost effective. This method can be used by the histology and pathology technicians and the researchers.

Histotecnología se refiere a la elaboración y preparación de los tejidos del cuerpo de tal forma que permitan un estudio satisfactorio de éste. El corte seccionado es una parte integral de la histología y la histopatología. Es un arte por sí mismo que requiere habilidad y conocimiento por parte del técnico o la persona que tiene que hacer el corte de la sección. En el método de rutina de la preparación de las secciones de parafina se encuentra a menudo la presencia de artefactos como arrugas finas o pliegues. Se han hecho intentos para eliminar las arrugas por flotación de las secciones en el baño de agua tibia. Sin embargo, este método no ha sido capaz de eliminar todas las arrugas de las secciones. Hemos diseñado un método sencillo y fiable, en el que las secciones de parafina fueron colocadas por flotación sobre alcohol etílico diluido con agua (1:15) antes de ser colocado en el baño de agua. A través de este método, hemos sido capaces de obtener secciones sin arrugas de calidad superior. La ventaja de nuestro método es que, es fácil de preparar el alcohol diluido y es rentable. Este método puede ser utilizado por los técnicos de histología, patología e investigadores.

Histomorfometria do corno uterino de gatas (Felis catus) submetidas à ovariosalpingohisterectomia / Histomorphometry of the uterine horn of queens (Felis catus) uterus submitted to ovaryhisterectomy

Dados histomorfométricos foram obtidos de cornos uterinos de gatas nulíparas (n=6), primíparas (n= 6), multíparalterações mais marcantes nas gatas tratadas; as variações morfológicas menos marcantes foram relativas ao diâmetro e epitélio glandulares; a presença de glândulas endometriais dilatadas foi encontrada somente nas gatas tratadas.

Histomorphometric data were obtained from cats uterine horns: either nulliparous (n = 6), primiparous (n = 6), multiparous (n = 6) and treated with contraceptive (n = 6). The material was collected after surgery, fixed in paraformaldehyde and embedded in paraplast® resin to be sliced in a microtome. The obtained sections were stained with hematoxylin -eosin and measured under a light microscope: uterine wall total thickness (μm), endometrium total height, endometrial glands diameter and glandular epithelia height, total myometrium, internal and external myometrium and vascular layer thickness. It was concluded that: 1 -contraceptive use and number of pregnancies altered the uterine structure, 2 -one pregnancy does not appear to affect the uterine lining structures as occurs in multiparous cats, 3 – there was no variation on the evaluated structures between nulliparous and primiparous cats except for inner myometrium, 4 - the total myometrium hight and the endometrium hight showed similar variations except for the contraceptive treated cats group, 5 - the outer myometrium showed marked changes in the treated cats, 6 – the less marked morphological variations were for the endometrial glands diameter and glandular epithelium hight, 7 - the presence of dilated endometrial glands was found only in treated cats.

Computerized three-dimensional reconstruction of cochlear microstructures / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore an useful and practical method for three-dimensional reconstruction of cochlear microstructure.@*METHOD@#A digital image dataset of magnified forty times of human cochlear tissue can be get from the serial sections of transverse cedukol images of male cadavers temporal bone. Photoshop 6.0 and Amira 3.0 software were used to deal with the digital image dataset, such as image-connecting and image inner-location and three-dimensional reconstruction of cochlear and scala media.@*RESULT@#The result of three-dimensional reconstruction of the cedukol images represents the anatomy of human cochlear and scala media transparently and objectively.@*CONCLUSION@#The three-dimensional reconstruction technique used in the study is practical for three-dimensional of inner ear microstructure.

Simulation and data analysis of stereological modeling based on virtual slices / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a computer-assisted stereological model for simulating the process of slice section and evaluate the relationship between section surface and estimated three-dimensional structure.</p><p><b>METHODS</b>The model was designed by mathematic method as a win32 software based on the MFC using Microsoft visual studio as IDE for simulating the infinite process of sections and analysis of the data derived from the model. The linearity of the fitting of the model was evaluated by comparison with the traditional formula.</p><p><b>RESULTS</b>The win32 software based on this algorithm allowed random sectioning of the particles distributed randomly in an ideal virtual cube. The stereological parameters showed very high throughput (>94.5% and 92%) in homogeneity and independence tests. The data of density, shape and size of the section were tested to conform to normal distribution. The output of the model and that from the image analysis system showed statistical correlation and consistency.</p><p><b>CONCLUSION</b>The algorithm we described can be used for evaluating the stereologic parameters of the structure of tissue slices.</p>

Retention and penetration of a conventional resin-based sealant and a photochromatic flowable composite resin placed on occlusal pits and fissures.

This study compares the retention and penetration of a conventional resin-based sealant (Fluroshield) and a photochromatic flowable composite resin (Tetric Flow Chroma) placed on occlusal pits and fissures and submitted to thermal or chemical cycling regimens. Penetration assessment--ten premolars were sealed with each material, isolated (except for the sealed surface) and immersed in 0.2% Rhodamine B. The teeth were serially sectioned in a mesiodistal direction. The images of the sections were digitized and analyzed (ImageLab). The distance between the most superficial and the deepest points on the occlusal central groove was calculated to determine the groove's total depth. The length of the central groove filled with the sealant was divided by its total depth to obtain the percentage of sealing of the occlusal groove. Retention assessment--30 premolars were sealed, their occlusal surfaces were photographed and the area occupied by the sealing materials was demarcated (ImageLab). The teeth were submitted to different treatments: thermocycled, stored in artificial saliva and immersed in acetic acid and saliva (10 cycles/day protocol for 30 days). New photographs were taken to assess the final area occupied by the materials. The difference between the final and initial area was calculated to obtain the material loss. The data was analyzed (two-way ANOVA and Tukey's test P<0.05). Both materials presented similar penetration of the occlusal central groove. After thermal and chemical cycling, the materials did not differ with respect to retention, except for immersion in acetic acid. In this case, Tetric Flow Chroma presented greater retention than Fluoroshield.

Study of methods of decalcification for making united slices of tooth and affiliated periodontic tissues / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To study the methods of decalcification for making united slices of tooth and affiliated periodontic tissues.</p><p><b>METHODS</b>Twenty-one samples containing dog molars and affiliated periodontic tissues were divided into seven mean groups. The pH value of solution, time of decalcification, weight and volume of samples, and content of decalcified calcium were detected. The slices were observed by HE, specific, and immunohistochemical stain.</p><p><b>RESULTS</b>The velocity of decalcification increased with decrease of solution pH. The weight of samples lightened by 37.61%, the volume reduced by 25.97% on average, and calcium decalcified was 174.49 mg per gram humid samples. The EDTA decalcification was slowest, but it was best. Decalcification was fast in Plank-Rycho solution while the section was worst, and faster in the formyl solution containing aluminium chloride than in EDTA, and the section was better.</p><p><b>CONCLUSIONS</b>The 50% formyl solution containing aluminium chloride is an ideal decalcifying solution.</p>

Technique of rat cochlea slicing and study of rat spiral ganglion neurons by infrared visual slice patch clamp method / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To establishing the cochlea slice technique and infrared visual slice patch clamp method in order to observe the electrophysiological characteristics of rat spiral ganglion neurons (SGN) METHODS: SD rats were divided into three groups according to postnatal days old (0-2 d, 3-6 d and 7-14 d). Making slice of SD rat cochlear quickly, using infrared differential interference contrast (IR-DIC) technique, together with slice patch clamp, the electrophysiological characteristics of rat spiral ganglion neurons were observed, and factors which affected the quality of cochlear slice and recording of patch clamp were analyzed.</p><p><b>RESULTS</b>The success rate of 3-6 days SD was the highest, and 2-4 pieces of slice could be made from each cochlea. Cochlea connecting with partial skull and integrity of cochlear hull were the key for making slice, and the angle of modiolus axis should be adjusted to be parallel to the knife and the preparing time should be shorter. The SGN cell of good condition could be easily found and the seal test became easier with the help of infrared visual slice patch clamp method. The rest membrane potential was (-45.6 +/- 5.3) mV (x +/- s, n=52) and the current of Na+ and K+ could be activated.</p><p><b>CONCLUSIONS</b>Cochlear slice technique can retain structural integrity, cell viability and their association in cochlea, which suggest that this technique provides carrier for electrophysiological study of rat spiral ganglion neurons, and patch clamp with infrared videomicroscopy method can be used to make direct real-time observation in electrophysiological experiments of SGN, which can provide important technique support and reference for deep study of electrophysiological characteristics of SGN and auditory neurotransmission in cochlea.</p>

Study with large slice technique and tissue microarray on lymph node involvement and micro-metastasis of rectal cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To study lymph node involvement and micro-metastasis of rectal cancer with large slice technique and tissue microarray.</p><p><b>METHODS</b>Large slice technique, combined with tissue microarray,was used in pathologic study of 31 patients after total mesorectal excision (TME) for rectal cancer.</p><p><b>RESULTS</b>Nine hundred and ninety- two lymph nodes were harvested and 148 were positive. More than 40% of positive lymph nodes were located in the outer layer of the mesorectum and in the same side of the mesorectum as the primary tumor was. Circumferential margin involvement was observed in 12 cases and correlated with the numbers of metastatic lymph nodes (Beta =1.166, P=0.041). Micrometastasis was found in 9 cases with negative pathological lymph nodes, but not correlated with tumor differentiation and stage (P> 0.05).</p><p><b>CONCLUSION</b>Large slice technique combined with tissue microarray facilitates the detection of lymph node involvement and micrometastasis. There is a predominance of lymph node metastasis in the outer layer and the same side of the mesorectum. Micrometastasis can be discovered in different stages of rectal cancer.</p>

Noncytopathic hepatitis A virus induces surface alterations in LLC-MK2 cells revealed by thin sections, negative staining, and scanning electron microscopy

Previous electron microscope studies of ultrastructural events during hepatitis A virus replication in experimentally infected cells have used only ultrathin section techniques. Nevertheless, no important differences were observed between infected and uninfected cells. This study was carried out using scanning electron microscopy and negative staining of whole LLC-MK2 cells grown directly on grids covered with support membranes, and then infected with an hepatitis A virus strain. Thin sections of infected and uninfected controls were also analyzed. An intricate web of projections forming a net between cell interfaces was observed only in infected cells. Some of these projections were more than 700 nm long and had ballooning tips. Nevertheless, HAV particles were not visualized in the infected cells.

Determination of the morphological irregularities in the middle and apical 1/3rd region of the root canal system of permanent maxillary incisors.

A thorough knowledge of the root canal anatomy and an understanding of its variations from the normal are mandatory for the successful root canal therapy. The assessment and exploration of the accessory canal and apical delta is necessary to combat the persistent infection at the periapical area. The purpose of this study is to determine the morphological irregularities in the middle and apical 1/3rd region of the root canal system of maxillary incisors. A total of 100 maxillary incisors were decalcified, processed, sectioned at the middle and apical 1/3rd region and observed under an ordinary microscope. The frequency of accessory canals, apical delta and type of canal configuration were studied. Accessory canals were found in 5% of the teeth. There was absence of apical delta in all the specimens. Total specimens showed single canal extended from the pulp chamber to the apex (Type 1 canal configuration).

Registration and 3D rendering of serial tissue section images / 生物医学工程学杂志

It is an important morphological research method to reconstruct the 3D imaging from serial section tissue images. Registration of serial images is a key step to 3D reconstruction. Firstly, an introduction to the segmentation-counting registration algorithm is presented, which is based on the joint histogram. After thresholding of the two images to be registered, the criterion function is defined as counting in a specific region of the joint histogram, which greatly speeds up the alignment process. Then, the method is used to conduct the serial tissue image matching task, and lies a solid foundation for 3D rendering. Finally, preliminary surface rendering results are presented.

Aspectos morfológicos das glândulas salivares de Lutzomyia (Lutzomyia) longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae: Phlebotoninae) em diferentes tempos pós-emergência e pós-repasto / Morphologic aspects of the glands salivares of Lutzomyia (Lutzomyia) longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae: Phlebotoninae) in different times after-emergency and after-repasto

Nos flebotomíneos, os estudos da morfologia interna, são esparsos; os que se referem às glândulas salivares (GS) são raros, antigos, e envolvem, predominantemente, as espécies do Velho Mundo(Adler & Theodor, 1926, Perfilev, 1928, Lewis, 1965).Este trabalho apresenta o estudo das (GS) da fêmea de Lutzomyia longipalpis, em cortes semifinos de espécimes incluídos em historesina em diferentes tempos pós-emergência e pós-repasto (corados pelo HE e pelo azul de bromofenol), e em órgãos dissecados em salina entomológica, corados pelo azul de tripan e observados ao microscópio óptico. As GS dessa espécie são órgãos pares de forma sacular, providos de dutos individuais rugosos que se unem para formar um duto comum, como descrito por outros autores em outras espécies. Neste estudo, GS globosas, com parede distendida, e apresentando na luz grânulos refratáves, foram observadas nos espécimes dissecados antes do repasto, e GS emurchecidas ou piriformes em espécimes analisados imediatamente após. Nos cortes histológicos dos insetos jovens, as GS se apresentaram pequenas, com envoltório formado por uma camada única de células epiteliais poliédricas apresentando núcleos e nucleolos pequenos, e contendo na luz grânulos grandes, basófilos, coráveis pelo azul de bromofenol. Estas células tornaram-se ativas 24 horas após a emergência, o que se evidenciou pelo aumento do volume nuclear e nucleolar, e pela presença de granulação fina na luz. Evidenciou-se um período de maturação da glândula nas primeiras 48 horas de vida, caracterizado por alterações no volume glandular, diminuição da espessura da parede e aumento de granulação proteíca que se torna mais fina com o tempo. Verificou-se ainda que, embora a alimentação sangüinea leve a alterações no volume (diminuição), forma (torna-se irregular e de parede espessa), e conteúdos glandulares, não se pode estabelecer um padrão para essas alterações devido grande variação encontrada nestes resultados nos diferentes tempos estudados. O achado de substância não protéica tanto nos cortes histológicos quanto nas GS dissecadas, e de gotículas não miscíveis no conteúdo glandular sugere a presença de lipídeos na saliva. Os possíveis mecanismos de ativação da glândula são discutidos e aspectos morfológicos de outros órgãos são apresentados.

Valor de la biopsia rápida en el diagnóstico de cancer / The accuracy of quick biopsy for cancer diagnosis

El objetivo fue estudiar estadísticamente el valor de la biopsia rápida para diagnosticar malignidad, global y específicamente en los diversos órganos y sistemas. Se consideró como presente la malignidad cuando la biopsia diferida así lo concluyó. Se realizó un estudio retropectivo de 659 biopsia rápida y sus correspondientes biopsias diferidas, efectuadas enel Servicio de Anatomía Patológica del Hosp. Dr. G. Fricke entre Septiembre de 1995 y Febrero de 1999.Seaplicó un test de screening para el análisis de los resultados al universo de muestras y por órganos y sistemas, excluyendo las biopsias rápidas no concluyentes