Comparación de las tinciones con hematoxilina y eosina versus azul de toluidina en la evaluación de los carcinomas cutáneos durante la cirugía micrográfica de Mohs / Comparative study of toluidine blue and hematoxylin and eosin staining in evaluating skin-carcinoma during Mohs micrographic surgery / Comparação da coloração de hematoxilina e eosina versus azul de toluidina na avaliação de carcinomas cutâneos durante a cirurgia micrográfica de Mohs

Resumen: Introducción: la cirugía micrográfica de Mohs es una técnica quirúrgica especializada para el tratamiento del cáncer de piel no melanoma. La histopatología cumple un rol fundamental, y la elección de la tinción es un punto de controversia. Objetivos: comparar el rendimiento de las tinciones de hematoxilina y eosina (HyE) versus azul de toluidina (AT) durante la cirugía. Método: estudio observacional, descriptivo y transversal a partir de noviembre de 2017 hasta mayo de 2018. Se incluyeron las láminas empleadas durante la cirugía en el período mencionado. Estas fueron analizadas por el cirujano de Mohs, tres residentes y una dermopatóloga. Se valoró el rendimiento de ambas tinciones, teniendo en cuenta las características celulares y los elementos del estroma. Resultados: se estudiaron 23 tumores (16 carcinomas basocelulares y 7 carcinomas espinocelulares). Al observarse al microscopio óptico tanto con la tinción de AT como con HyE no se encontraron diferencias significativas entre ambos grupos en lo global, sólo en algunas características, especialmente con la HyE. Conclusiones: es el primer trabajo en Uruguay que compara la eficacia de las dos tinciones durante la cirugía micrográfica de Mohs. Como conclusión tanto la tinción de HyE como el AT son muy buenas técnicas para el diagnóstico de carcinomas cutáneos.

Abstract: Introduction: Mohs micrographic surgery is a specialized surgical technique used to treat nonmelanoma carcinoma. Histopathology plays a vital role in the diagnosis of this condition, and the choice staining method is controversial. Objective: to compare results in the use of hematoxylin and eosin (H&E) versus Toluidine blue (TB) staining during surgery. Method: observational, descriptive and transversal study conducted from November, 2017 until May, 2018 of the slides used during surgeries in the selected period. Slides were analysed by the Mohs surgeon, 3 residents and a dermopathologist to evaluate the results of both staining methods, in consideration of cell features and stromal elements. Results: 23 tumors were analysed (16 Basal Cell carcinomas and 7 Squamous Cell Carcinoma). Microscopic observation of slides prepared with Toluidine blue and hematoxylin and eosin stains did not show significant global differences between both groups, except in terms of a few characteristics, in particular with hematoxylin and eosin stains. Conclusions: this was the first study in Uruguay to evaluate the effectiveness of both staining methods during Mohs micrographic surgery, and it concluded that both Toluidine blue and hematoxylin and eosin stains are very good techniques in evaluating skin-cancer.

Resumo: Introdução: a cirurgia micrográfica de Mohs é uma técnica cirúrgica especializada para o tratamento do câncer de pele não melanoma. A histopatologia desempenha um papel fundamental, onde a escolha da coloração é um ponto de controvérsia. Objetivos: comparar o desempenho das colorações de hematoxilina e eosina versus azul de toluidina durante a cirurgia. Método: estudo observacional, descritivo e transversal de novembro de 2017 a maio de 2018. Foram incluídas as lâminas utilizadas durante as cirurgias no referido período. Estas foram analisadas pelo cirurgião especializado na técnica de Mohs, 3 residentes e um dermatopatologista onde foi avaliado o desempenho de ambas as colorações, levando em consideração as características celulares e os elementos do estroma. Resultados: foram estudados 23 tumores (16 carcinomas basocelulares e 7 carcinomas espinocelulares). Quando observados ao microscópio de luz para coloração AT e H&E, não foram encontradas diferenças significativas entre os dois grupos em geral, apenas em algumas características, especialmente com o H&E. Conclusões: é o primeiro estudo no Uruguai que compara a eficácia dos 2 corantes durante a cirurgia micrográfica de Mohs. Em conclusão, tanto a coloração com hematoxilina e eosina quanto com azul de toluidina são técnicas muito boas para o diagnóstico de carcinomas de pele.

Comparación de métodos histoquímicos para el análisis histológico de muestras de encía humana / Comparison of histochemical methods for the histological analysis of human gingival samples

Introducción: Las técnicas de coloración histológica son útiles en el análisis ultraestructural de muestras tisulares, incluyendo el tejido gingival. Objetivo: Comparar la utilidad de tres métodos histoquímicos (hematoxilina-eosina, Masson Goldner y rojo sirio) en la identificación de elementos celulares y otros constituyentes del tejido gingival. Métodos: Estudio experimental in vitro que comprendió el análisis de tejidos gingivales de donantes sanos sin signos de inflamación gingival y con indicación de cirugía periodontal. Las muestras de encía se obtuvieron mediante gingivectomía, se procesaron e incluyeron en parafina, posteriormente se realizaron cortes con un micrótomo y se depositaron en portaobjetos de adhesión con polisina. Las muestras se agruparon y fueron teñidas con hematoxilina-eosina, Masson Goldner y rojo sirio, finalmente fueron visualizadas en un microscopio óptico Leica DM 750®. La lectura de los hallazgos fue realizada por patólogos orales. Resultados: La coloración hematoxilina-eosina evidencia elementos celulares y extracelulares del tejido epitelial y conectivo. Núcleos de color azul violeta, citoplasmas rosados, fibras de colágeno de matiz rosa claro, arteriolas y vénulas con túnica adventicia, media e íntima diferenciadas. La coloración Masson Goldner diferencia núcleos de coloración púrpura y citoplasma fucsia, presenta especificidad en identificar fibras de colágeno con tonalidad verde, distribuidas densa, homogénea y paralelamente en el tejido conectivo gingival. La tinción rojo sirio, permitió identificar las fibras de colágeno de color rosa brillante, mientras que el tejido epitelial y los vasos sanguíneos fueron de color amarillo. Conclusión: Cada coloración histológica evaluada en el presente trabajo tiene cierta afinidad y sensibilidad por estructuras celulares y componentes de la matriz extracelular específica. Su empleo es útil en el estudio de tejidos gingivales y podrían contribuir en el análisis de biopsias gingivales(AU)

Introduction: Histological staining techniques are useful in the ultrastructural analysis of tissue samples, including gingival tissue. Objective: Compare the usefulness of three histochemical methods (hematoxylin-eosin, Masson-Goldner and sirius red) for identification of cellular elements and other constituents of gingival tissue. Methods: An in vitro experimental study was conducted which included the analysis of gingival tissue from healthy donors without gingival inflammation signs and indication of periodontal surgery. The gum samples were obtained by gingivectomy, processed with paraffin, cut with a microtome and placed on Polysine adhesion slides. The samples were grouped, stained with hematoxylin-eosin, Masson Goldner and sirius red, and visualized under a Leica DM 750® microscope. Reading of the findings was performed by oral pathologists. Results: Hematoxylin-eosin staining found cellular and extracellular elements of the epithelial and connective tissue: violet-blue nuclei, pink cytoplasms, light rose collagen fibers, and arterioles and venules with differentiated tunica adventitia, media and intima. Masson-Goldner staining differentiated purple nuclei and fuchsia cytoplasm. It displayed specificity identifying green collagen fibers with dense, homogeneous and parallel distribution in the gingival connective tissue. Sirius red staining allowed identification of bright rose collagen fibers, whereas epithelial tissue and blood vessels were yellow. Conclusion: Each of the histological staining methods evaluated in the study shows a certain affinity with and sensitivity to cellular structures and components of the specific extracellular matrix. All three are useful for the study of gingival tissue and could contribute to the analysis of gingival biopsies(AU)

Value of fluorescence staining in histopathological diagnosis of Acanthamoeba keratitis / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To investigate the application value of fluorescent staining technique in the detection of amoebic pathogens in corneal tissue biopsy, and to apply fluorescent staining technique in the histopathological diagnosis of Acanthamoeba keratitis(AK), comparing the results with those of hemotoxyiln-eosin staining(HE staining)and periodic acid-schiff staining(PAS staining), and analyzing the sensitivity and specificity of these three staining methods.<p>METHODS:Specimens of infected corneal tissue were collected from 74 cases(75 eyes), and then they were divided into an AK group and a non-Acanthamoeba keratitis(NAK)group based on the results of corneal scraping, culture and histopathological diagnosis. The tissues of consecutive sections were stained with HE staining, PAS staining and fluorescence respectively, and the sensitivity and specificity of the three staining methods for the diagnosis of AK were analyzed. Area under the curve(AUC)was calculated using the receiver operating characteristic(ROC)curve. Further analysis was performed to count the number of Acanthamoeba pathogens found by the three staining methods under the same magnification field of view at the same site, and to clarify the diagnostic value of fluorescent staining technique for AK.<p>RESULTS: The sensitivity of HE staining was 69%(27/39)with a specificity of 92%; the sensitivity of PAS staining was 62%(24/39)with a specificity of 97%, and the sensitivity of fluorescent staining was 95%(37/39)with a specificity of 97%. There were differences in the sensitivity of the three staining methods for the diagnosis of AK(χ2=19.857, <i>P</i><0.001), and pairwise comparison revealed that the differences between HE staining and fluorescent staining, PAS staining and fluorescent staining for the diagnosis of AK were statistically significant(<i>P</i>=0.003,<0.001), while the difference in sensitivity between HE staining and PAS staining for the diagnosis of AK was not statistically significant(<i>P</i>=0.978). The maximum AUC was 0.960 for fluorescence staining, followed by 0.804 for HE staining and 0.794 for PAS staining, respectively. The median number of amoeba cysts detected by HE staining, PAS staining and fluorescent staining at the same site under the same magnification field of view was 4(0, 11), 2(0, 9)and 12(3, 33), respectively(χ2=56.561, <i>P</i><0.001). Pairwise comparison revealed that the differences in the number of amoeba cysts found by HE staining and fluorescence staining, PAS staining and fluorescence staining were statistically significant(<i>P</i><0.001), while the difference in the number of amoeba cysts found by HE staining and PAS staining was not statistically significant(<i>P</i>=0.210). Fluorescently stained histopathological sections make it easier to identify amoebic pathogens.<p>CONCLUSION:Fluorescent staining technique is more sensitive to histopathological diagnosis of AK than HE staining and PAS staining, which can significantly improve the positive rate of detection of amoebic pathogens.

MicroRNA-141-3p expression in serum of patients with cerebral hemorrhage and its mechanism / 解剖学报

Objective To analyze the expression level of microRNA-141-3p (miR-141-3) in patients with intracerebral hemorrhage (ICH), and explore the effect and mechanism of miR-141-3p on cerebral hemorrhage injury in rats. Methods Forty patients with ICH and 40 healthy controls in total were enrolled in this study. The expression of miR- 141-3p in peripheral blood serum was determined by the Real-time PCR method. The target relationship between miR-141- 3p and NOD-like receptor 3 (NLRP3) 3′ UTR was confirmed by dual luciferase reporter assay. miR-141-3p agonist and agonist NC were injected into rats via the lateral ventricle, respectively. On day 7 after treatment, the neurological function score was evaluated, and then all rats were killed to obtain brain tissue. Brain water content was examined by the dried and wet mass. HE staining was conducted to observe the pathological changes of cerebral tissue. The mRNA expressions of NLRP3 and miR-141-3p were detected by Real-time PCR. The protein expression of interleukin (IL)-lβ, IL-6 and IL-18 were detected by Western blotting analysis. Results The expression of miR-141-3p in serum of ICH patients was significantly down-regulated compared to healthy controls and negatively correlated with the severity of edema around the hematoma [(0.068±0.038) vs (0.520±0.028), t = 15.93, P<0.001; r =-0.8948, -0.9434 to-0.8087, P<0.001 ]. The result of luciferase reporter assay showed that miR-141-3p was related to the regulation of NLRP3 gene expression. The relative expression levels of miR-141-3p in agonist group were significantly higher than those in the agonist NC group (P< 0.001), while the expression levels of NLRP3, IL-lβ, IL-6 and IL-18 were significantly lower than those in the agonist NC group (P< 0.001). Compared with the agonist NC group, the cerebral water content reduced significantly (P< 0.001), and the neurological function score was significantly improved on the day 7 after treatment in agonist group (P< 0.001). The result of HE staining showed that injection of miR-141-3p in ICH rats could reduced the severity of edema around the hematoma. Conclusion MiR-141-3p alleviates ICH-induced inflammatory injury in rat possibly by modulating miR-141-3p.

Comparison of the apoptotic effects of topically applied papaverine, diltiazem, and nitroprusside to internal thoracic artery

Abstract Objective: To detect and to compare the apoptotic effects of intraoperatively topically applied diltiazem, papaverine, and nitroprusside. Methods: Internal thoracic artery segments of ten patients were obtained during coronary bypass grafting surgery. Each internal thoracic artery segment was divided into four pieces and immersed into four different solutions containing separately saline (Group S), diltiazem (Group D), papaverine (Group P), and nitroprusside (Group N). Each segment was examined with both hematoxylin-eosin and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method in order to determine and quantify apoptosis. Results: Apoptotic cells were counted in 50 microscopic areas of each segment. No significant difference was observed among the four groups according to hematoxylin-eosin staining. However, the TUNEL method revealed a significant increase in mean apoptotic cells in the diltiazem group when compared with the other three groups (Group S=4.25±1.4; Group D=13.31±2.8; Group N=9.48±2.09; Group P=10.75±2.37). The differences between groups were significant (P=0.0001). No difference was observed between the samples of the diabetic and non-diabetic patients in any of the study groups. Conclusion: The benefit of topically applied vasodilator drugs must outweigh the potential adverse effects. In terms of apoptosis, diltiazem was found to have the most deleterious effects on internal thoracic artery graft segments. Of the analyzed medical agents, nitroprusside was found to have the least apoptotic activity, followed by papaverine. Diabetes did not have significant effect on the occurrence of apoptosis in left internal thoracic artery grafts.

Comparison of HE and Giemsa staining in diagnosis of bacterial and allergic conjunctivitis in children / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To explore the diagnostic effect of hematoxylin-eosin staining(HE)and Giemsa staining in the diagnosis of bacterial and allergic conjunctivitis in children. <p>METHODS:Totally 422 children with conjunctivitis diagnosed by conjunctivitis from the ophthalmology department of our hospital during 2016-10/2019-10 as the research objects. HE and Giemsa staining methods were used to stain the conjunctival scratches, and the staining results were used to diagnose bacterial/allergic conjunctivitis. Observe the positive detection rate of the two staining results for bacterial/allergic conjunctivitis and the staining situation. <p>RESULTS: The positive rate(33.0%)and coincidence rate(63.6%)of HE staining for the diagnosis of bacterial conjunctivitis were significantly lower than Giemsa staining(90.7% and 88.8%, <i>P</i><0.001), while the positive rate of allergic conjunctivitis was not significantly different 90.8% <i>vs </i>87.2%, <i>P</i>>0.05).<p>CONCLUSION: The Giemsa staining method can accurately diagnose bacterial conjunctivitis in children and the method is simple. Both HE and Giemsa staining methods have good diagnostic effects on allergic conjunctivitis, which can provide a basis for improving the clinical diagnosis efficiency and early treatment options.

Effect of miR-155 on acute lung injury in burned rats: Changes of nuclear factor-kappa B pathway / 中国组织工程研究

BACKGROUND: Currently, studies have focused on the role and mechanism of nuclear factor-kappa B pathway in the pathological process of acute lung injury in burned rats, such as the targeting inhibition of kB kinase by miR-155, which further weakens the activity of nuclear factor-KB and plays a role in acute lung injury in burned rats. However, there are still some pathological mechanisms to be studied and confirmed. OBJECTIVE: To investigate the effect of miR-155 on acute lung injury in burned rats through nuclear factor-KB pathway. METHODS: The rat models of acute lung injury were established by warm water bath simulating bum injury. The burned rats were divided into acute lung injury, miR-155-mimics and miR-155-inhibitor groups. After fluid resuscitation, the rats in the miR-155-mimics and miR-155-inhibitor groups were injected into the tail vein of 5 |_iL of miR-155-mimics and miR-155-inhibitions, respectively. The expression levels of tumor necrosis factor-a and interleukin-1 p in bronchoalveolar lavage fluid were detected by ELISA. The lung morphology in the three groups was observed by hematoxylin-eosin staining. The protein expression levels of nuclear factor-KB and cyclooxygenase 2 were evaluated by western blot assay. The nuclear factor-KB protein in lung tissues was detected by immunohistochemistry. RESULTS AND CONCLUSION: (1) The results of hematoxylin-eosin staining showed that the severity of lung injury in the miR-155-inhibitor group, acute lung injury group and the miR-155-mimics group was increased gradually (P < 0.05). (2) ELISA results showed that compared with the acute lung injury group, the expression levels of tumor necrosis factor-a and interleukin-1 p were increased in the miR-155-mimics group (P < 0.05), and decreased in the miR-155-inhibitor group (P < 0.05). (3) Western blot assay results showed that compared with the acute lung injury group, the expression levels of nuclear factor-KB and cyclooxygenase 2 proteins were increased in the miR-155-mimics group (P < 0.05), and decreased in the miR-155-inhibitor group (P < 0.05). (4) Immunohistochemical results showed that the expression level of nuclear factor-KB was increased in the miR-155-inhibitor group, which was dark brown. The expression of nuclear factor-KB in cytoplasm and nucleus of neutrophils, mononuclear macrophages, alveolar epithelial cells was the most obvious. (5) These results indicate that in lung tissue cells, decreased miR-155 can down-regulate nuclear factor-KB activity, which reduces the inflammatory response of the lung between the damaged tissue. The study was approved by the Laboratory Animal Ethics Committee of the First People’s Hospital of Neijiang, approval No. 1801270.

Lycium barbarum polysaccharide intervenes with CD151 and matrix metalloproteinase 3 in the articular cartilage in a rabbit model of knee osteoarthritis / 中国组织工程研究

BACKGROUND: Studies have shown that Lycium barbarum polysaccharide (LBP) has the functions of anti-aging, nerve protection, anti-fatigue, blood sugar control, anti-oxidation, and anti-tumor. It may have some protective effects against osteoarthritis of the knee, but have been rarely reported. CD151 and matrix metalloproteinase 3 (MMP-3) are two common cytokines for assessing knee osteoarthritis. OBJECTIVE: To observe the effect of LBP on the expression of CD151 and MMP-3 in rabbit osteoarthritis. METHODS: Sixty-four healthy 6-month-old white rabbits were randomly divided into four groups: blank group, model group, LBP group and normal saline group. Animal models of knee osteoarthritis were made using Hulth method in the rabbits except those in the blank group. The rats in the LBP and normal saline groups were fed with normal dose of LBP and normal saline for 4 weeks, and then the articular cartilage tissues were taken from the affected side at 12 weeks after modeling. The morphological changes of the articular cartilage were observed by hematoxylin-eosin staining. The expression levels and spatial distribution of CD151 and MMP-3 in articular cartilage was observed by immunohistochemical staining and western blot. Ethic approval was given by the People’s Hospital of Ningxia Hui Autonomous Region (approval No. 2014-30817). RESULTS AND CONCLUSION: immunohistochemistry staining and western blot results showed that the absorbance values and protein expression of MMP-3 and CD-151 were significantly lower in the LBP group than the normal saline and model groups (P < 0.05). Therefore, the expression of CD151 and MMP-3 in the articular cartilage of osteoarthritis was increased, and LBP could inhibit the expression of CD151 and MMP-3 in osteoarthritis, so as to slow down the occurrence of osteoarthritis.

Improvement program on pretreatment of acid decalcified tissue in hematoxylin-eosin staining / 华西口腔医学杂志

OBJECTIVE@#To explore the treatment conditions of acid decalcified specimens and improve the poor quality of sections and unclear structure of hematoxylin-eosin (HE) staining caused by the change in pH in tooth and hard tissue after acid decalcification.@*METHODS@#A total of 20 cases of oral pathological specimens that contain hard tissues were decalcified and treated with routine treatment, concentrated ammonia water immersion treatment, and saturated lithium carbonate solution immersion treatment. The quality and HE staining effects of hard tissue sections treated with different methods were compared.@*RESULTS@#Compared with routine treatment, lithium carbonate saturated solution treatment showed complete sections. Hematoxylin is strongly stained, the nucleus is clear, and the cytoplasm is bright.@*CONCLUSIONS@#Soaking acid decalcified specimens in lithium carbonate saturated solution before embedding in dehydration can neutralize the acidic environment of the tissue. The quality of sections and HE staining effect are improved and are suitable for the pretreatment of acid decalcified tissue samples of oral pathology.

Effect of spinal cord decellularized scaffold on the repair of spinal cord defects in rats / 解剖学报

Objective Exploring the effect of spinal cord decellularized scaffold on spinal cord defects and observing the behavior and regeneration of rats after operation. Methods The spinal cords of 30 SD rats were treated with 3% Triton X-100 and 2% sodium deoxycholate on oscillator. The cell residue and the spatial structure of the tissue were compared before and after treatment, in order to understand the tissue structure of the stent itself. 90 SD rats were randomly divided into control group, simple injury group and stent transplantation group. Excision of the spinal cord 9-10 segments in the simple injury group and the stent graft group the acellular scaffold was transplanted to the stent graft group. Behavioral scores were observed postoperatively. At 4, 8, and 12 weeks, the spinal cords of the injured part of the rats were taken for HE staining and immunofluorescence detection of nerve regeneration-related proteins. Results After decellularization of the spinal cord, the nerve cells and axons were completely removed, and the extracellular matrix of the spinal cord was preserved. Scanning electron microscopy revealed that the scaffold retained a certain porous network scaffold structure. In the experiment of decellularized scaffold in vivo, the Basso-Beattie-Bresnahan(BBB) score showed that the recovery of hindlimb motor function in rats with decellularized scaffolds was better than that in rats with simple injury. HE staining showed that the decellularized scaffold could fill the defect of the spinal cord segment and accelerate the repair process of the injured spinal cord. Immunofluorescence showed that there was a certain axonal regeneration in the injured part of the stent transplantation group. Conclusion The spinal cord decellularized scaffold retains the extracellular matrix and has a certain spatial structure, which can accelerate the process of spinal cord defect repair to a certain extent, and has a certain promoting effect on nerve regeneration.

Identificación de Helicobacter pylori por medio de la coloración especial de Warthin-Starry en biopsias de pacientes con gastritis crónica folicular, previamente negativas en la tinción de hematoxilina-eosina / Warthin-Starry stain identification of Helicobacter pylori in biopsies of patients who previously tested negative in hematoxylin-eosin staining for follicular gastritis

Resumen Las técnicas empleadas para la detección del Helicobacter pylori (H. pylori) son no invasivas e invasivas. En estas últimas, la presencia del H. pylori se determina a partir de la tinción de hematoxilina-eosina (HE), prueba rutinaria, mientras que en pocas ocasiones se aplica la tinción de Warthin-Starry (WS) como coloración especial. Objetivo: identificar la presencia de H. pylori por medio de la coloración especial de la WS en biopsias de pacientes con gastritis crónica folicular, previamente negativas en la tinción HE. Materiales y métodos: se desarrolló un estudio de tipo descriptivo transversal, en un período de 12 meses. Se tomaron los bloques de parafina de las muestras de la mucosa gástrica de pacientes con diagnóstico de gastritis crónica e hiperplasia folicular. Además, se extrajo un corte histológico del mismo bloque, al cual se le aplicó HE y se determinó la presencia o ausencia de H. pylori. Así, de estar ausente, se tomó del mismo bloque un corte adicional y se aplicó WS. Esto se evaluó con el fin de identificar la existencia o no del bacilo. Resultados: se recolectaron 314 muestras; 209 fueron negativas y 105 fueron positivas para HE. El 45 % (94) de estas muestras fueron positivas respecto a la presencia del bacilo, al aplicar la segunda coloración, y el 55 % (115) de las muestras persistieron negativas. Conclusión: el hallazgo de H. pylori es significativamente alto al aplicar la coloración de WS a muestras cuyo estudio histológico evidenció la ausencia del bacilo en biopsias de la mucosa gástrica, especialmente en muestras con escasa cantidad de bacterias.

Abstract Non-invasive and invasive techniques can be used for detection of Helicobacter pylori. An invasive technique identifies the bacteria through routine hematoxylin-eosin staining. Warthin-Starry stain is rarely used. Objective: Our objective was to identify H. pylori by Warthin-Starry staining of patient's biopsies with chronic follicular gastritis who had previously tested negative in hematoxylin-eosin staining. Materials and methods: This is a descriptive, cross-sectional descriptive study that was carried out over a period of 12 months. The study examined paraffin blocks of samples taken from the gastric mucosa of patients diagnosed with chronic gastritis and follicular hyperplasia. A histological section was extracted from a block and tested with hematoxylin-eosin staining for the presence or absence of H. pylori. If absent, an additional cut was taken from the same block and Warthin-Starry staining was used to retest for the presence of the bacteria. Results: Of the 314 samples collected, 209 tested negative, and 105 tested positive for H. pylori when hematoxylin-eosin staining was used. Of the 209 negative samples, 45% (94) tested positive when Warthin Starry stain was used, and 55% (115) still tested negative. Conclusion: Findings of H. pylori are significantly higher when Warthin Starry stain was used to test samples whose previous histological study had evidenced an absence of the bacillus, especially in samples with a small amount of bacteria.

Oral Administrations of Hancornia speciosa Gomes Latex Do Not Increase Bone Neoformation / Administrações Orais do Látex da Hancornia speciosa Gomes não aumentam a neoformação óssea

Abstract Objective The present work aimed to evaluate the systemic effect of H. speciosa latex on bone neoformation. Methods For this, the latex was collected and diluted to 3% and 50%. A total of 28 Wistar rats were submitted to surgery to create a 5 mm diameter defect in the parietal bone. This experiment was conducted in 2 different periods: 1 and 2. For each period, the rats were divided into 3 groups: Control Group, Latex3 Group, and Latex50 Group, which received, respectively, daily administrations of 0.5 mL of distilled water, latex to 3% and latex to 50% by gavage, orally. The rats of periods 1 and 2 were euthanized, respectively, 15 and 30 days after the surgery, and the calvaria was collected. The results were analyzed using the ANOVA and Tukey tests; the significance level was 0.05. Results We show that, in each analyzed period, the experimental groups had the same amount of newly formed bone in the calvaria defect. Conclusion We conclude that daily and oral administrations of H. speciosa latex to 3% and to 50% over a period of 15 and 30 days does not contribute to the increase of the area of the newly formed bone in the calvaria defect.

Resumo Objetivo Este trabalho objetivou avaliar o efeito sistêmico do látex de H. speciosas obre a neoformação óssea. Métodos Para isso, o látex foi coletado e diluído a 3% e a 50%. Um total de 28 ratos Wistar foi submetido a cirurgia para a criação de um defeito de 5 mm de diâmetro no osso parietal. Esse experimento foi conduzido em dois períodos distintos: 1 e 2. Para cada período, os ratos foram divididos em 3 grupos: Grupo Controle, Grupo Látex3 e Grupo Látex50 que receberam, respectivamente, administrações diárias de 0,5 mL de água destilada, látex a 3% e látex a 50% por gavagem, via oral. Os ratos dos períodos 1 e 2 foram eutanasiados, respectivamente, 15 e 30 dias após a cirurgia e a calvária foi coletada. Os resultados foram analisados utilizando os testes ANOVA e Tukey; o nível de significância estabelecido foi 0,05. Resultados Mostramos que, em cada período analisado, os grupos experimentais tiveram a mesma quantidade de osso neoformado no defeito da calvária. Conclusão Portanto, concluímos que administrações diárias e orais do látex de H. speciosa a 3% e a 50% durante um período de 15 e 30 dias não contribui para o aumento da área do osso neoformado no defeito da calvária.

Tetrachromic VOF/Masson's trichrome/H and E stains: Unmasking their usability in differential stromal hard tissue staining

Background: Identification and differentiation of stromal hard tissue components is a challenging task. Numerous methods of demonstrating these components have been worked upon in the past. Although some of the methods have been successful, there are many drawbacks of employing them. The need of the hour, therefore is to develop and use a simple, rapid and cost-effective method of identifying stromal hard tissues as they may signify an important change in the diagnosis of the pathology. Our aim is therefore to observe the usability of tetrachromic VOF stain over Hematoxylin and Eosin and Masson's Trichrome in routinely encountered head and neck pathologies. Materials and Method: Routine cases such as Central and peripheral ossifying fibromas, osteomas, giant cell granulomas, osteomyelitis and malignancies like osteosarcomas were retrieved from the department archives and 3 sections from each block were prepared to stain with H and E, Masson's trichrome and modified tetrachromic VOF stains respectively using standard staining protocol. Results: Tetrachromic VOF takes an upper hand in stromal hard tissue differentiation irrespective of the pathology. Conclusion: Modified tetrachromic VOF is simple, cost-effective method and can be employed for diagnosis of cases with hard tissue differentiation within the stroma on routine basis.

Effect and mechanism of essential oil from Zanthoxylum bungeanum in microemulsion gel preparation on percutaneous delivery of complex components / 中国中药杂志

This study aimed to investigate the transdermal enhancing effect of essential oil from Zanthoxylum bungeanum(Z. bungeanum oil) in microemulsion gel(ZO-ME-gel) on permeation of different components,and reveal the transdermal enhancing mechanism of ZO-ME-gel. A series of components with different log P values were selected as model drugs and encapsulated in ZO-ME-gel to simplify and characterize the complex components of traditional Chinese medicine. The transdermal behavior of the model drugs was further examined using the improved Franz diffusion cell method. Then attenuated total reflection Fourier transform infrared spectroscopy(ATR-FTIR),differential scanning calorimetry(DSC) studies and hematoxylin-eosin(HE) staining were used to investigate the effects of Z. bungeanum oil and ZO-ME-gel on keratin,intercellular lipids and microstructure of the stratum corneum(SC). The results showed that Z. bungeanum oil and ZO-ME-gel had a good transdermal enhancing effect on both hydrophilic and lipophilic drugs,and the best effect was achieved when log P value was-0. 5. The transdermal enhancing mechanism of Z. bungeanum oil and ZO-ME-gel was related to affecting the order of the SC lipids,changing lipid fluidity and protein conformation,and disrupting the integrity of the SC structure. 5% Z. bungeanum oil had greater transdermal enhancing effect and destruction of SC structure than ZO-ME-gel. These results suggested that Z. bungeanum oil loaded in microemulsion gel still had a good transdermal enhancing effect although the effect was not as great as Z. bungeanum oil itself,in addition,ZO-ME-gel was less irritating to the skin and safer to use,which had a guiding role in the development and clinical application of Z. bungeanum oil-containing traditional Chinese medicine topical preparations.

Comparing The Efficacy Of Dishwash Solution, Diluted Lemon Water, Coconut Oil And Xylene As Deparaffinizing Agents For Hematoxylin And Eosin Staining Procedure

Aim: To compare the efficacy of dish wash solution, diluted lemon water, coconut oil and xylene as deparaffinizingagents for hematoxylin and eosin staining procedure.Objective: The objective is to find eco-friendly deparaffinizing agents like dish wash solution, diluted lemonwater and coconut oil as substitute to xylene and comparing the staining characteristics of each individualdeparaffinizing agent with Xylene.Materials and Methods: The study comprised of paraffin embedded 45 blocks of various tissues. Each block offour sections of 5 microns thickness was prepared. They were considered in four different groups like A, B, C andD. Tissue sections in Group A were stained with H & E method where xylene was used as deparaffinizing agent. Theother three sections were stained with H & E where dish wash solution, diluted lemon water and coconut oil wereused as deparaffinising agent’s alternative to Xylene. Staining characteristics were compared with xylene andscoring was given. The total score of 3–5 was regarded as satisfactory for diagnosis and less than that isinsufficient for diagnosis.Statitistical Analysis: Chi square test was used.Results: Adequacy of staining characteristics such as nuclear, cytoplasm, uniformity, clarity and crispiness ofstaining for diagnosis was greater with dish wash solution followed by diluted lemon water, coconut oil andxylene.Conclusion: The Eco-Friendly deparaffinizing agents such as dish wash solution, diluted lemon water, and coconutoil can be used as alternatives to xylene

Protective Effect and Metabonomics Research of "Biao-Ben Acupoint Combination" Electroacupuncture in Chronic Myocardial Ischemia Model Rats / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) on ischemic electrocardiogram (ECG), histopathological changes and serum metabolite profile in chronic myocardial ischemia (CMI) rats, so as to reveal its mechanisms underlying protecting ischemic myocardium. METHODS: A total of 45 male Wistar rats were randomly divided into normal control, CMI model and EA groups, with 15 rats being in each group. The rats in the control group received subcutaneous injection of 0.9% normal saline (5 mg•mg－1•d－1, for 7 days), and those in the model and EA groups received subcutaneous injection of isopropylarterenol hydrochloride (5 mg•mg－1•d－1, for 7 days) to establish CMI model. EA (2 Hz/100 Hz, 1 mA) was applied to bilateral "Zusanli" (ST 36), "Guanyuan" (CV 4) and "Neiguan" (PC 6) for 10 min, once daily for 21 days. The ECG-ST segment of the standard limb lead II was used for evaluating the severity of myocardial ischemia, and the histopathological changes of myocardium were observed under microscope after H．E. staining. The profile of serum metabolites was analyzed by nuclear magnetic resonance mass spectrometry combined with principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) in 21 rats (n＝7 in each group)．. RESULTS: After modeling, the amplitude of ECG-ST was significantly increased in comparison with the normal control group (P 1). The PLS-DA analysis revealed deviations in 51 differential biomarkers in serum，among which, Glucose, Lactate, Creatine, Acetate and 3-Hydroybutyrate may contribute to the effect of EA in improving CMI. CONCLUSION: EA stimulation of acupoints can ameliorate ischemic myocardial injury in CMI rats, which may be related to its effect in regulating serum sugar, lipid metabolism and energy metabolism.

Evaluation of cell proliferation and endometrial thickness of bitches in different periods of diestrus

ABSTRACT This study aimed to determine the histological features of the endometrium of bitches, as well as the cell proliferation at specific moments of diestrus, 10, 20, 30, 40, 50 and 60 days post ovulation, correlating the endometrial thickness with the uterine cell proliferation and the metabolic state (weight, blood glucose and plasma cholesterol) of the animals. Therefore, the right and left uterine horns of 26 clinically healthy bitches submitted to ovariohysterectomy were histologically analyzed 10, 20, 30, 40, 50 and 60 days post ovulation. The hematoxylin-eosin and AgNOR staining techniques were performed. All parameters were evaluated by ANOVA and post-hoc Tukey test (p<0.05). The correlation between endometrial thickness and uterine cell proliferation, weight, blood glucose and plasma cholesterol of animals was observed using the Pearson method (p<0.05). In the present study, it is concluded that endometrial thickness does not differ at any of the moments analyzed in diestrus. The endometrial thickness is not influenced by hormones, weight, blood glucose or serum cholesterol of bitches in this phase of the estrous cycle. However, there is greater cell proliferation in the endometrium at day 40 compared to day 60 post ovulation under the influence of the endocrine profile.

Experimental study on regulating effect of protosappanin A on dendritic cell maturation and functional state / 心血管康复医学杂志

Objective: To explore regulating effect of protosappanin A (PrA) on dendritic cell (DC) maturation.Methods: SPF male Wistar rats and SD rats were selected as subjects.During DC maturation induced by lipopolysaccharide (LPS), methyl thiazolyl tetrazolium (MTT) method was used to screen proper concentrations of PrA.Different concentrations of PrA were used to pretreat LPS-induced DC, difference of DC surface molecule CD80 and CD86 expressions were analyzed;DC's ability in activating T cell proliferation, expression levels of CD4, CD25 and Foxp3 on surface of regulatory T cells (Treg), and levels of interleukin (IL)-10 and IL-12 secreted by DC in supernatant were measured.Results: Compared with immature DC (imDC) group, there were significant rise in expressions of DC surface molecule CD80[(31.50±29.04)% vs.(63.80±14.03)%] and CD86[(36.10±27.21)% vs.(62.60±12.37)%] in LPS-DC group, P<0.01 both;compared with LPS-DC group, there were significant reductions in expressions of CD80[(63.80±14.03)% vs.(39.70±26.60)] and CD86[(62.60±12.37)% vs.((37.90±26.93)] in 20-DC group (DC cultured with 20nmol/L PrA), P<0.05 both.Compared with LPS-DC group, there were significant reductions in DC-activated T cell proliferation capacity [(0.39±0.06) vs.(0.32±0.46) vs.(0.28±0.08)] and IL-12 level [(250.00±89.81) pg/ml vs.(176.80±49.89) pg/ml vs.(134.30±60.64) pg/ml], and significant rise in expression of Treg [(0.42±0.23) vs.(0.76±0.20) vs.(0.93±0.52)] and IL-10 level [(145.80±70.28) pg/ml vs.(274.00±131.93) pg/ml vs.(354.00±146.22) pg/ml] in 5-DC group and 20-DC group (P<0.05 all for 5-DC group, P<0.01 all for 20-DC group).Conclusion: PrA can inhibit LPS-induced DC maturation, including reducing expressions of surface molecule CD80 and CD86, suppressing the capacity to activate allogeneic T lymphocyte proliferation, inducing Treg proliferation and affecting levels of relative cytokines.

Morphological characteristics of expressions of DKK1 in different periods of tooth development in postnatal mice and their significances / 吉林大学学报(医学版)

Objective:To study the expression characteristics of Dickkopf1 (DKK1) in different time and space during tooth development of the postnatal mice, and to provide the theoretical basis for clarifying the mechanism of Wnt signaling pathway in regulating the tooth development.Methods:The postnatal Kunming mice at days 0.5, 6.5, 12.5, 18.5, 24.5, and 30.5 respectively after birth were selected and divided into various groups by time,three in each group.The mice in each group were sacrificed and the paraffin sections of mandibular bone including the first molar were prepared at the thickness of 5 μm, followed by HE staining and immunohistochemical staining in order to detect the expressions of DKK1 in tooth tissue and periodontal tissue.Results:At 0.5 d after birth, the mandibular first molar tooth germ was in the bell stage.At 6.5 d the enamel development of mandibular first molar was almost completed, and the epithelium root sheath extended to the root direction.At 12.5 d the dentin development of crown was completed, with the root formatted about 1/3. At 18.5 d the root had formatted about 2/3.At 24.5 d the root had reached the full length.At 30.5 d the apical foramen was narrow, and the root development was basically completed.There was no DKK1 expression at 0.5 d, but it expressed in the odontoblasts and predentin at 6.5 d. From days 12.5 to 30.5,the expressions of DKK1 were positive in periodontal ligament, alveolar bone, and cellular cementum as odontoblasts, which were gradually increased with the prolongation of time.However, no expression of DKK1 was detected in the pulp.Conclusion:DKK1 shows regular expressions at different tooth developmental stages after birth, suggesting its potential role in the growth of dentin and periodontal tissues.

Apoptosis in oral epithelial dysplastic lesions and oral squamous cell carcinoma: A prognostic marker.

Background: Apoptotic index (AI) using light microscopy as an indirect measure to assess the significance of apoptosis as a proliferative marker in dysplastic lesions and malignant epithelial lesions of the oral cavity. Aims: (1) To quantify the apoptotic bodies/cells in oral epithelial dysplastic (OED) lesions and oral squamous cell carcinoma (OSCC). (2) To measure AI in OED and OSCC. (3) To compare AI in OED and OSCC. Settings and Design: The proposed laboratory‑based retrospective study involved the use of hematoxylin and eosin (H and E)‑stained slides of previously diagnosed OED lesions and OSCC from institutional archives. Materials and Methods: This study constituted 50 cases, each of H and E‑stained slides of previously diagnosed cases of OED and OSCC. AI was calculated as the number of apoptotic bodies/cells expressed as a percentage of the total number of nonapoptotic tumor/dysplastic cells counted in each case. Statistical Analysis Used: Nonparametric tests such as Kruskal–Wallis test and Mann–Whitney test were used. Results: There was a statistically significant increase in AI from OED to OSCC (P = 0.000). Conclusions: Further studies need to be undertaken to detect and understand the apoptotic mechanisms in the progression from OED to OSCC.