Development of flow cytometry and its application in plant research / 生物工程学报

Flow cytometry is a multi-parameter, rapid and efficient method for qualitative analysis and quantitative determination of various fluorescently labeled particles in liquid flow. Flow cytometry has been applied in multiple disciplines such as immunology, virology, molecular biology, cancer biology and infectious disease monitoring. However, the application of flow cytometry in plant research is hampered due to the special composition and structure of plant tissues and cells, such as cell walls and secondary metabolites. In this paper, the development, composition and classification of flow cytometry were introduced. Subsequently, the application, research progress and application limitations of flow cytometry in plant field were discussed. At last, the development trend of flow cytometry in plant research was prospected, which provides new perspectives for broadening the potential application scope of plant flow cytometry.

Caracterización morfométrica nuclear de glándulas mamarias sanas en mujeres adultas mayores

Introducción: Las glándulas mamarias son órganos que durante las diferentes etapas de la vida en la mujer sufren modificaciones, donde se involucran los procesos de proliferación, diferenciación y apoptosis, bajo el control hormonal. Sin embargo, una vez que cesan dichas influencias hormonales ocurren cambios que llevan a la involución de dicho órgano. Objetivo: Caracterizar el factor de forma, perímetro, área y volumen de los núcleos de las células epiteliales glandulares mamarias. Métodos: Para caracterizar las glándulas mamarias sanas en mujeres de 60 años y más, se realizó un estudio de serie de casos en 14 mujeres fallecidas que no tenían lesiones benignas o malignas del órgano. Todas examinadas por el departamento de Anatomía Patológica del Hospital Provincial Vladimir Ilich Lenin en Holguín, en el período comprendido de septiembre 2018 a septiembre 2019. Para mejor valoración, la muestra de estudio se dividió en dos grupos de edades: de 60-75 años de edad y mayores de 75 años. Resultados: Tanto el factor de forma como el perímetro, área y volumen de los núcleos de las células epiteliales de los conductos mamarios son menores en las mujeres mayores de 75 años. Conclusiones: Existen diferencias notables en los indicadores morfométricos estudiados en ambos grupos de edades. Específicamente el tamaño y la forma de los núcleos de células epiteliales se ven afectados con la edad, lo cual se corresponde con la baja actividad metabólica de las células epiteliales mamarias en esta etapa de la vida.

Introduction: The mammary glands are organs that during the different stages of life in women undergo modifications, where the processes of proliferation, differentiation and apoptosis are involved, under hormonal control. However, once these hormonal influences cease, changes occur that lead to the involution of said organ. Objective: To determine the shape factor, perimeter, area and volume of the nuclei of glandular epithelial cells. Methods: To characterize healthy mammary glands in women aged 60 years and older, a case series study was conducted on 14 deceased women who had no benign or malignant lesions of the organ. All examined by the Department of Pathological Anatomy of the Provincial Hospital V.I. Lenin in Holguín, in the period between September, 2018 - September, 2019. For a better assessment, the study sample was divided into two age groups: from 60 to 75 years of age; age and older than 75 years. Results: Both the shape factor and the perimeter, area and volume of the nuclei of the epithelial cells of the mammary ducts are lower in women older than 75 years. Conclusions: There are notable differences in the morphometric indicators studied. Epithelial cell nuclei are affected with age, which corresponds to the low metabolic activity of mammary epithelial cells at this stage of life.

Genotoxicity in the oral cells of older people from a Brazilian rural area: a population-based study

Abstract The purpose of this population-based, observational, and cross-sectional study was to evaluate alterations in the oral cells of a population of older people from a Brazilian rural area, using the micronucleus technique to investigate possible associated genotoxic factors. A questionnaire was applied and clinical examination and collection of oral mucosal cells were performed for all older people (≥ 60 years) from a town in southern Brazil. Demographic and socioeconomic variables, deleterious habits (drinking and tobacco use), presence of gastro-oesophageal reflux disease (GERD), and the use of proton pump inhibitors (PPIs) were considered the exposure variables, whereas metanuclear changes (MCs) and the prevalence of cell micronuclei (MN) were considered outcomes. Out of 489 older people, 447 were included in the study, among whom 50.8% were men with a mean age of 70.9 years and 83.9% had a monthly family income greater than US$ 500.00. GERD symptoms were present in 36.2% of the individuals, and 29.1% used PPIs daily, 53.3% consumed alcoholic beverages, and 46.7% used tobacco. The analysis of 1,000 oral mucosal cells per subject showed a MN frequency of 0-2 per individual, and MCs were detected with an average of 15 units per individual (median = 11 per individual). Poisson regression did not show statistical association between the exposure variables and the outcomes (presence of MN and MCs), except for the use of PPIs, which was a protective factor for the prevalence of MN [PR 0.6 (CI 0.3-0,9)]. Age, sex, family income, tobacco use and drinking, and GERD were not associated with the number of MN and MCs in oral mucosal cells of the investigated older people.

Effect of synthetic peptide cSN50.1 on the malignant behavior of hepatocellular carcinoma HepG2 cells and its mechanism / 临床肝胆病杂志

ObjectiveTo investigate the effect of cSN50.1 on the proliferation， migration， invasion， and colony formation of HepG2 cells and its mechanism. MethodsHepG2 cells were divided into cSN50.1 0 μmol/L， cSN50.1 10 μmol/L， cSN50.1 30 μmol/L， cSN50.1 50 μmol/L， cSN50.1 70 μmol/L， and cSN50.1 90 μmol/L groups， and CCK－8 assay was used to investigate the effect of different concentrations of cSN50.1 on the proliferation of HepG2 cells and calculate half－maximal inhibitory concentration （IC50）. HepG2 cells were divided into cSN50.1 0 μmol/L， cSN50.1 10 μmol/L， cSN50.1 30 μmol/L， and cSN50.1 50 μmol/L groups， and wound healing assay， Transwell assay， and colony－forming assay were used to investigate the effect of different concentrations of cSN50.1 on the migration， invasion， and colony formation of HepG2 cells. HepG2 cells were divided into Control group， SP600125 group （an inhibitor of the AP－1 signaling pathway）， and cSN50.1 group to investigate the influence of the AP－1 signaling pathway on the effect of cSN50.1 on hepatocellular carcinoma cells， and RT－PCR and Western Blot were used to measure the expression of CXCL5， TNF－α， and c－Jun protein in cytoplasm and nucleus. HepG2 cells were divided into Control group， PDTC group （an inhibitor of the NF－κB signaling pathway）， and cSN50.1 group to investigate the influence of the NF－κB signaling pathway on the effect of cSN50.1 on hepatocellular carcinoma cells， and RT－PCR and Western Blot were used to measure the expression of CXCL5， TNF－α， and NF－κB protein in cytoplasm and nucleus. A one－way analysis of variance was used for comparison between multiple groups， and the SNK－q test was used for further comparison between two groups. ResultsCompared with the 0 μmol/L group， the 10 μmol/L group had no significant changes in proliferation， migration， invasion， and colony formation abilities （P >0.05）； the 30 μmol/L group had no significant change in proliferation ability （P>0.05）， but with significant reductions in migration， invasion， and colony formation abilities （P<0.05）； the 50 μmol/L group had significant reductions in proliferation， migration， invasion， and colony formation abilities （all P<0.01）； the 70 μmol/L and 90 μmol/L groups had a significant reduction in cell proliferation ability （P<0.01）， but with a cell survival rate of below 50%. Compared with the Control group， the SP600125， PDTC， and cSN50.1 groups had significant reductions in the mRNA and protein expression levels of CXCL5 and TNF－α （all P<0.05）. Compared with the Control group， the SP600125 group， the PDTC group， and the cSN50.1 group had a significant reduction in nuclear protein of c－Jun and NF－κB expression （P<0.05）； the SP600125 group and the PDTC group had a significant reduction in cytoplasmic protein of c－Jun and NF－κB expression （P<0.05）； the cSN50.1 group had a significant increase in cytoplasmic protein of c－Jun and NF－κB expression （P<0.05）. ConclusionThis study shows that cSN50.1 can inhibit the malignant behavior of hepatocellular carcinoma cells and reduce the expression of CXCL5 and TNF－α by inhibiting the nuclear import of c－Jun and NF－κB in hepatocellular carcinoma cells.

Follicular Lesions with Papillary Nuclear Characteristics: Differences in Chromatin Detected by Computerized Image Analysis

ABSTRACT Objective: Follicular lesions of the thyroid with papillary carcinoma nuclear characteristics are classified as infiltrative follicular variant of papillary thyroid carcinoma-FVPTC (IFVPTC), encapsulated/well demarcated FVPTC with tumour capsular invasion (IEFVPTC), and the newly described category "non-invasive follicular thyroid neoplasm with papillary-like nuclear features" (NIFTP) formerly known as non-invasive encapsulated FVPTC. This study evaluated whether computerized image analysis can detect nuclear differences between these three tumour subtypes. Materials and methods: Slides with histological material from 15 cases of NIFTP and 33 cases of FVPTC subtypes (22 IEFVPTC, and 11 IFVPTC) were analyzed using the Image J image processing program. Tumour cells were compared for both nuclear morphometry and chromatin textural characteristics. Results: Nuclei from NIFTP and IFVPTC tumours differed in terms of chromatin textural features (grey intensity): mean (92.37 ± 21.01 vs 72.99 ± 14.73, p = 0.02), median (84.93 ± 21.17 vs 65.18 ± 17.08, p = 0.02), standard deviation (47.77 ± 9.55 vs 39.39 ± 7.18; p = 0.02), and coefficient of variation of standard deviation (19.96 ± 4.01 vs 24.75 ± 3.31; p = 0.003). No differences were found in relation to IEFVPTC. Conclusion: Computerized image analysis revealed differences in nuclear texture between NIFTP and IFVPTC, but not for IEFVPTC.

Naturally derived sulforaphene inhibits proliferation of mantle cell lymphoma by down-regulating CRM1 expression / 肿瘤

Objective: To investigate the expression level of chromosomal region maintenance 1 (CRM1) in mantle cell lymphoma, and to explore the possible mechanism of naturally derived sulforaphene LFS-01 inhibiting the proliferation of mantle cell lymphoma cells. Methods: The expression of CRM1 in mantle cell lymphoma was analyzed by Oncomine data mining. After treatment with LFS-01, the viability of mantle cell lymphoma JeKo-1 cells was detected by CCK-8 assay, the nuclear transport function of CRM1 protein was measured by laser confocal microscopy, the expression of CRM1 protein was detected by Western blotting, the cell cycle arrest and apoptosis were analyzed by FCM and transmission electron microscopy, the expression change of apoptosis pathway-related proteins was detected by Western blotting, and the impact of caspase inhibitor z-VAD-FAM on the effects of LFS-01 was detected by CCK-8 assay and FCM. Lentiviral infection was used to establish a stable JeKo-1 cells expressing mutant CRM1 (C528S), then the effects of LFS-01 on the nuclear transport function of CRM1 and the proliferation of JeKo-1 cells were detected by laser confocal microscopy and CCK-8, respectively. The transcriptional level in JeKo-1 cells after LFS-01 treatment was detected by RNAseq, and the viability of JeKo-1 cells treated with Toll-like receptor (TLR) inhibitor TAK-242 and LFS-01 was measured by CCK-8 assay. Results: CRM1 was overexpressed in mantle cell lymphoma (P < 0.05). LFS-01 inhibited the proliferation of JeKo-1 cells, and the 50% inhibitory concentration (IC50) at 24 h and 48 h were 5.81 μmol/L and 9.09 μmol/L, respectively. 20.0 μmol/L LFS-01 inhibited the nuclear transport function of CRM1 (P < 0.05), 6.0 μmol/L LFS-01 down-regulated the expression level of CRM1 (P < 0.05), 10.0 μmol/L LFS-01 induced cell cycle arrest at G2/M phase and induced apoptosis (both P < 0.05), and 4 μmol/L LFS-01 up-regulated the expression levels of poly ADP-ribose polymerase (PARP), cleaved caspase-3 and cleaved caspase-9 (all P < 0.01). The caspase inhibitor z-VAD-FAM reversed the apoptosis-induction effect of LFS-01 (P < 0.01). CRM 1 mutation eliminated the effects of LFS-01 on CRM1 function and cell proliferation (both P < 0.05). LFS-01 inhibited the cell proliferation-related pathways (P < 0.01), and TLR inhibitor TAK-242 combined with LFS-01 had synergetic inhibitory effect on JeKo-1 cells (P < 0.01). Conclusion: LFS-01 can inhibit the growth of mantle cell lymphoma cells through inducing cell cycle arrest and apoptosis, and CRM1 is essential in this process. In addition, LFS-01 and TLR inhibitor TAK-242 have synergetic inhibitory effect on mantle cell lymphoma cells.

Effect of transplantation of human amniotic mesenchymal stem cells on the lung tissue pathologic change in paraquat induced pulmonary fibrosis rats / 实用医学杂志

Objective To investigate the impact of transplantation of human amniotic mesenchymal stem cells(hAMSCs)on the histopathological change in paraquat-induced pulmonary fibrosis in rats. Methods Forty-six female SD rats were randomly divided into the sham surgery group and the hAMSCs transplant group. Pulmonary fibrosis model was induced by 2％ of paraquat intragastric administration(100 mg/kg/rat). hAMSCs were injected through caudal vein(2 × 106 cells/mL/rat). The histopathological changes were observed through microscopy after HE and the immunohistochemical staining. Results General conditions in rats received hAMSCs transplantation were better than those of the model rats. More large area and white fibrosis nidus were observed in bilateral lung of model rats,with less dispersal spot or nidus. The construction of lung tissue was disordered in the model rats. The thickness of alveolar wall was found increased. There were large area interstitial hyperplasia and a large number of inflammatory cells infiltrations. The construction of lung tissue was apparently improved. A majority of alveolar wall was monolayer cell. There were only less and small area with interstitial hyperplasia. Inflammatory cell infiltration was significantly decreased. The anti-human nucleus specific antibody positive hAMSCs were observed planted and survived in lung interstitial tissue. And few hAMSCs were observed planted in alveolar wall. Conclusion The transplanted hAMSCs can be planted and survived in lung tissue ,and may play a therapeutic role in araquat-induced pulmonary fibrosis.

Effect of transplantation of human amniotic mesenchymal stem cells on the lung tissue pathologic change in paraquat induced pulmonary fibrosis rats / 实用医学杂志

Objective To investigate the impact of transplantation of human amniotic mesenchymal stem cells(hAMSCs)on the histopathological change in paraquat-induced pulmonary fibrosis in rats. Methods Forty-six female SD rats were randomly divided into the sham surgery group and the hAMSCs transplant group. Pulmonary fibrosis model was induced by 2％ of paraquat intragastric administration(100 mg/kg/rat). hAMSCs were injected through caudal vein(2 × 106 cells/mL/rat). The histopathological changes were observed through microscopy after HE and the immunohistochemical staining. Results General conditions in rats received hAMSCs transplantation were better than those of the model rats. More large area and white fibrosis nidus were observed in bilateral lung of model rats,with less dispersal spot or nidus. The construction of lung tissue was disordered in the model rats. The thickness of alveolar wall was found increased. There were large area interstitial hyperplasia and a large number of inflammatory cells infiltrations. The construction of lung tissue was apparently improved. A majority of alveolar wall was monolayer cell. There were only less and small area with interstitial hyperplasia. Inflammatory cell infiltration was significantly decreased. The anti-human nucleus specific antibody positive hAMSCs were observed planted and survived in lung interstitial tissue. And few hAMSCs were observed planted in alveolar wall. Conclusion The transplanted hAMSCs can be planted and survived in lung tissue ,and may play a therapeutic role in araquat-induced pulmonary fibrosis.

Herramienta alternativa para la clasificación de células cervicales utilizando solo rasgos del núcleo / Alternative tool for classification of cervical cells using only features of the nucleus

El cáncer de cérvix uterino representa una de las mayores amenazas de muerte por cáncer entre las mujeres. Con el avance continuo en la medicina y la tecnología, las muertes por esta enfermedad han disminuido significativamente. Las investigaciones referentes a este tema han podido determinar síntomas claves que permiten detectar a tiempo esta enfermedad para darle un tratamiento oportuno. La citología convencional es una de las técnicas más utilizadas, siendo ampliamente aceptada, de bajo costo, y con mecanismos de control. Con el objetivo de aliviar la carga de trabajo a los especialistas, algunos investigadores han propuesto el desarrollo de herramientas de visión computacional para detectar y clasificar las transformaciones en las células de la región del cuello uterino. La presente investigación tiene como objetivo proveer a los investigadores de una herramienta de clasificación automática, aplicable a las condiciones existentes en los centros médicos y de investigación del país. Esta herramienta debe ser capaz de clasificar las células del cuello del útero, basándose solamente en las características extraídas de la región del núcleo y sin utilizar las características del citoplasma, de manera que se reduzca la tasa de falsos negativos en la prueba de Papanicolaou. A partir del estudio realizado, se obtuvo una herramienta haciendo uso de la técnica k-vecinos más cercanos con la distancia manhattan, el cual mostró un alto desempeño manteniendo valores de AUC superiores al 91 por ciento y llegando hasta un 97.1 por ciento con respecto a los clasificadores SVM y RBF Network, los que también fueron analizados(AU)

Cervix cancer is one of the biggest threats of cancer death among women. With continued advances in medicine and technology, deaths from the disease have fallen significantly. The investigations concerning this issue have determined key symptoms to detect the disease in time to give timely treatment. Conventional cytology is one of the most widely used techniques, being widely accepted, inexpensive, and with control mechanisms. In order to alleviate the workload of specialists, some researchers have proposed the development of computer vision tools to detect and classify the changes in the cells of the cervical region. This research aims to provide a tool for automatic classification, applicable to medical conditions and research centers of the country. This tool should be able to classify the cells of the cervix, based solely on the features extracted from the core region without using the characteristics of the cytoplasm, so that the rate of false negative Pap test is reduced. From the study, a tool is obtained using the k nearest-neighbors manhattan distance technique, which showed a high performance maintaining AUC values greater than 91 percent and reaching 97.1 percent over classifiers SVM and RBF Network, which were also analyzed(AU)

Clinical and pathologic features of small gastrointestinal stromal tumor / 中华消化内镜杂志

Objective To investigate the clinical and pathologic features of small gastrointestinal stromal tumors (small GISTs,d ＜ 2.0 cm).Methods Medical records of 95 patients undergoing surgery (endoscopic surgery,thoracoscopic/laparoscropic surgery and open surgery)and diagnosed as having GISTs by pathology and immunohistochemistry in Nanfang hospital from October 2003 to June 2014 were retrospectively analyzed.Based on clinical and pathological results,correlation analyses between risk factors for endoscopic ultrasonography(EUS) and Mitotic count(MI),clinicopathologic parameter and NIH risk classification were performed.Results Among 95 cases (104 lesions),88 were single,while 7 were multiple;81.7％ (85/104) small GISTs arose from stomach,including 87.1％ (74/85)in middle-upper stomach;5 cases (5.3％) presented calcification of different degrees,3 cases(3.2％) presented local necrosis and 2 cases (2.1％) with arrangement of epithelioid cells;88 cases (92.6％) were very low grade of NIH risk classification,6 cases (6.3％) were intermediate risk and 1 case(1.1％) was high risk.Positive rates of CD34 and CD117 were 95.8％ (91/95) and 96.8％ (92/95) respectively.The risk factors (border,mucosal surface,echo and heterogeneity) of EUS had no correlation with mitotic count(P＞0.05).The correlation analysis between clinicopathologic features and NIH risk classification revealed tumors more than 1.5 cm had a striking correlation with NIH risk classification (P＜ 0.05).Conclusion Most small GISTs,single or multiple,located at middle-upper stomach,were of very low or low risk,and have a favorable prognosis.But it has worse biological behavior and a higher grade risk when the diameter is more than 1.5 cm,intervention should be recommended.

Difference of the Nuclear Green Light Intensity between Papillary Carcinoma Cells Showing Clear Nuclei and Non-neoplastic Follicular Epithelia in Papillary Thyroid Carcinoma

BACKGROUND: There is subjective disagreement regarding nuclear clearing in papillary thyroid carcinoma. In this study, using digital instruments, we were able to quantify many ambiguous pathologic features and use numeric data to express our findings. METHODS: We examined 30 papillary thyroid carcinomas. For each case, we selected representative cancer cells showing clear nuclei and surrounding non-neoplastic follicular epithelial cells and evaluated objective values of green light intensity (GLI) for quantitative analysis of nuclear clearing in papillary thyroid carcinoma. RESULTS: From 16,274 GLI values from 600 cancer cell nuclei and 13,752 GLI values from 596 non-neoplastic follicular epithelial nuclei, we found a high correlation of 94.9% between GLI and clear nuclei. GLI between the cancer group showing clear nuclei and non-neoplastic follicular epithelia was statistically significant. The overall average level of GLI in the cancer group was over two times higher than the non-neoplastic group despite a wide range of GLI. On a polygonal line graph, there was a fluctuating unique difference between both the cancer and non-neoplastic groups in each patient, which was comparable to the microscopic findings. CONCLUSIONS: Nuclear GLI could be a useful factor for discriminating between carcinoma cells showing clear nuclei and non-neoplastic follicular epithelia in papillary thyroid carcinoma.

Parámetros morfométricos de la mucosa en pacientes portadores de leucoplasia bucal con displasia epitelial / Morphometric parameters of the mucosa in patients that carry oral leukoplakia with epithelial dysplasia

Fundamento: la leucoplasia bucal es la lesión premaligna más frecuente, puede presentarse con displasia epitelial, que es el rasgo más importante de sus diversos cambios histopatológicos. Los métodos morfométricos y estereológicos permiten complementar el análisis histopatológico convencional de la displasia epitelial. Objetivo: caracterizar las lesiones de la mucosa con leucoplasia bucal según grado de displasia epitelial a través de indicadores morfométricos y estereológicos. Método: se realizó un estudio observacional descriptivo. El universo lo conformaron 68 biopsias de pacientes con diagnóstico clínico e histopatológico de leucoplasia y la muestra fue de 15 biopsias de leucoplasias displásicas, en las que se analizaron parámetros morfométricos y estereológicos. Resultados: se encontró que la altura de las crestas epiteliales y la altura del epitelio aumentan a medida que avanza el grado de la displasia epitelial. El valor promedio y su desviación estándar del factor de forma en la displasia ligera fue de 0,86 ± 0,08, en la displasia moderada fue de 0,88 ± 0,09 y en la displasia severa fue de 0,85 ± 0,11. El promedio y la desviación estándar del volumen nuclear y la densidad de perfiles nucleares epiteliales descienden al aumentar la displasia. Conclusiones: a partir del análisis de las variables analizadas en esta investigación podemos sugerir el estudio y la evaluación de la posibilidad de la existencia de algún modelo de relación volumen nuclear-perfiles nucleares-altura del epitelio como instrumento para determinar el comportamiento de la displasia.

Background: oral leukoplakia is the most frequent premalignant lesion; it can appear with epithelial dysplasia that is the most important feature of its various histopathological changes. Morphometric and stereological methods allow complementing the conventional histopathological analysis of epithelial dysplasia. Objective: to characterize the lesions of the mucosa with oral leukoplakia according to the degree of epithelial dysplasia through morphometric and stereological indicators. Method: a descriptive, observational study was conducted. The universe was composed of 68 biopsies from patients with a histopathological and clinical diagnosis of leukoplakia; the sample was composed of 15 biopsies from patients with dysplastic leukoplakias in which morphometric and stereological parameters were analyzed. Results: it was found that as the degree of epithelial dysplasia becomes greater, the more the height of the epithelial crest and the height of the epithelium rise. The average value and its standard deviation of the shape factor in minor dysplasia was of 0.86 ± 0.08, in moderate dysplasia was of 0.88 ± 0.09, and in serious dysplasia was of 0.85 ± 0.11. As the dysplasia becomes greater, the average and the standard deviation of the nuclear volume and the density of epithelial nuclear profile decrease. Conclusions: from the analysis of the variables through this investigation, we can suggest the study and evaluation of the possibility of the existence of a model of nuclear volume-nuclear profile-height of the epithelium relation, as an instrument to determine the behavior of dysplasia.

Estudio histomorfométrico del núcleo celular del carcinoma papilar de tiroides / Histomorphometric study of the cellular nucleus of the thyroid papillary carcinoma

Se realizó una evaluación morfométrica del carcinoma papilar tiroideo, con el objetivo de identificar posibles indicadores morfométricos que contribuyan a precisar su diagnóstico histológico. Se estudiaron 32 biopsias diagnosticadas en el Hospital Universitario “Celestino Hernández Robau”, de Santa Clara. Se determinaron las variables morfométricas nucleares: área, perímetro, radio y factor de forma, utilizando el programa de computación COMSDI PLUS versión 1.0. El carcinoma papilar en sus variantes hísticas no clásicas (folicular, oxífila y esclerosante difusa) presentaron valores más elevados del área, perímetro y radio nuclear, mientras que el carcinoma papilar, en su variante clásica, mostró un valor mayor de factor de forma. El estudio de correlación entre las variables estudiadas, en ambas variantes hísticas, no mostró similitud en sus resultados.

A morphometric evaluation of thyroid papillary carcinoma was carried out, with the objective of identifying possible morphometric parameters that contribute to precise its histological diagnosis. A number of 32 diagnosed biopsies of the University Hospital “Celestino Hernández Robau”, from Santa Clara were studied. Nuclear morphometric variables were determined, such as, area, perimeter, range and shape factor, by means of COMSDI PLUS version 1.0 computing program. Papillary carcinoma in its non classical histological variants (follicular, oxyphilic and diffuse sclerosing) showed more elevated values in area, perimeter and nuclear range, while, papillary carcinoma, in its classical variant, showed a high value in shape factor. The correlation study between studied variables did not showed similarity in its results in both histological variants.

Location of recombinant adenovirus Ad5/F35-HF2S and its interaction with Bcr-Abl in chronic myeloid leukemia K562 cells / 肿瘤

Objective: To testify the location of recombinant adenovirus Ad5/F35-HF2S in chronic myeloid leukemia K562 cells and its interaction with the fusion protein Bcr-Abl. Methods: The recombinant adenovirus Ad5/F35-HF2S was infected into K562 cells, then the expression of fusion protein HF2S in K562 cells was assessed by RT-PCR (reverse transcription-PCR) and Western blotting. The location of HF2S protein in K562 cells was demonstrated by immunofluorescence and Western blotting assay. The immunofluorescence was performed to verify the co-localization of HF2S and Bcr-Abl. The co-immunoprecipitation assay was employed to detect the interaction between HF2S and Bcr-Abl. Results: The leukemia cell line K562 was effectively infected by the adenovirus Ad5/F35-HF2S. The fusion protein HF2S and Bcr-Abl were co-located and interacted with each other in the cytoplasm. Conclusion: HF2S can be successfully expressed and interacted with Bcr-Abl in the cytoplasm of K562 cells. These results provide the basement of the targeted therapy for chronic myeloid leukemia. Copyright © 2013 by TUMOR.

Interaction of Hsc73 with CXCR4 in nuclei of renal cell carcinoma A498 cells / 第二军医大学学报

Objective: To investigate the role of protein Hsc73 (heat shock cognate protein 73 000) in renal cell cancer metastasis promoted by SDF-1/CXCR4 axis, so as to determine whether Hsc73 participates in CXCR4 nuclear localization. Methods: Western blotting analysis was used to observe the expression of Hsc73 in A498 cells over-expressing CXCR4. The location of Hsc73 and the interaction of CXCR4 with Hsc73 were investigated in SDF-1-stimulated A498 cells by immunohistochemical staining, Co-IP (Co-Immunoprecipitation) experiment, etc. Results: Hsc73 was up-regulated in A498 cells over-expressing CXCR4. Hsc73 was mainly found in the cytoplasm of A498 cells; after stimulation with SDF-1, some Hsc73 appeared in the nuclei. Hsc73 protein was found in the nuclei of A498 cells after Co-IP with anti-CXCR4 antibody. Conclusion: Hsc73 as a common molecular chaperone participates in the intra-cellular translocation of CXCR4; Hsc73 also plays a key role in the activation of SDF-1/CXCR4 signal pathway and may be involved in the nuclear translocalization of CXCR4.

Memantine prevents cardiomyocytes nuclear size reduction in the left ventricle of rats exposed to cold stress

OBJECTIVES: Memantine is an N-methyl-d-aspartate (NMDA) glutamate receptor antagonist used to treat Alzheimer's disease. Previous studies have suggested that receptor blockers act as neuroprotective agents; however, no study has specifically investigated the impact that these drugs have on the heart. We sought to evaluate the effects of memantine on nuclear size reduction in cardiac cells exposed to cold stress. METHOD: We used male EPM-Wistar rats (n=40) divided into 4 groups: 1) Matched control (CON); 2) Memantine-treated rats (MEM); 3) Rats undergoing induced hypothermia (IH) and 4) Rats undergoing induced hypothermia that were also treated with memantine (IHM). Animals in the MEM and IHM groups were treated by oral gavage administration of 20 mg/kg/day memantine over an eight-day period. Animals in the IH and IHM groups were submitted to 4 hours of hypothermia in a controlled environment with a temperature of - 8ºC on the last day of the study. RESULTS: The MEM group had the largest cardiomyocyte nuclear size (151 ± 3.5 μm³ vs. CON: 142 ± 2.3 μm³; p<0.05), while the IH group had the smallest mean value of nuclear size. The nuclear size of the IHM group was preserved (125 ± 2.9 μm³) compared to the IH group (108 ± 1.7 μm³; p<0.05). CONCLUSION: Memantine prevented the nuclear size reduction of cardiomyocytes in rats exposed to cold stress.

Modulation of the caveolin-3 localization to caveolae and STAT3 to mitochondria by catecholamine-induced cardiac hypertrophy in H9c2 cardiomyoblasts

We investigated the effect of phenylephrine (PE)- and isoproterenol (ISO)-induced cardiac hypertrophy on subcellular localization and expression of caveolin-3 and STAT3 in H9c2 cardiomyoblast cells. Caveolin-3 localization to plasma membrane was attenuated and localization of caveolin-3 to caveolae in the plasma membrane was 24.3% reduced by the catecholamine-induced hypertrophy. STAT3 and phospho-STAT3 were up-regulated but verapamil and cyclosporin A synergistically decreased the STAT3 and phospho-STAT3 levels in PE- and ISO-induced hypertrophic cells. Both expression and activation of STAT3 were increased in the nucleus by the hypertrophy. Immunofluorescence analysis revealed that the catecholamine-induced hypertrophy promoted nuclear localization of pY705-STAT3. Of interest, phosphorylation of pS727-STAT3 in mitochondria was significantly reduced by catecholamine-induced hypertrophy. In addition, mitochondrial complexes II and III were greatly down-regulated in the hypertrophic cells. Our data suggest that the alterations in nuclear and mitochondrial activation of STAT3 and caveolae localization of caveolin-3 are related to the development of the catecholamine-induced cardiac hypertrophy.

Cold stress effects on cardiomyocytes nuclear size in rats: light microscopic evaluation / Efeitos do estresse pelo frio sobre o tamanho nuclear do cardiomiócito em ratos: avaliação por microscopia de luz

INTRODUÇÃO: Hipotermia corporal induzida e resfriamento do miocárdio são métodos efetivos em relação à proteção domiocárdio durante cirurgias cardíacas e isquemia. É descrito na literatura que a exposição a temperaturas extremamente baixas causa comprometimentos de miofilamentos e de cristas mitocondriais em cardiomiócitos, entretanto, nenhum estudo analisou os efeitos do estresse pelo frio no tamanho do núcleo dos cardiomiócios. OBJETIVOS: Analisar os efeitos do estresse agudo pelo frio sobre o tamanho do núcleo dos cardiomiócitos. MÉTODOS: O estudo foi realizado em ratos Wistar adultos, pesando 300-310g (n=20). Os ratos foram divididos em dois grupos: 1) Controle (CON) e; 2) Hipotermia induzido (IH). Os animais do grupo IH foram expostos a uma temperatura controlada de -8ºC, durante 4 horas uma única vez. Foi realizada análise histológica de fígados e glândulas adrenais para examinar a condição de estresse. O tamanho do núcleo dos cardiomiócitos foi examinado por três investigadores independentes com o mesmo critério padronizado e posteriormente analisado pelo coeficiente de correlação de Bartko (R>0,75=concordância positiva). Teste t de Student foi aplicado. O nível de significância foi considerado como P<0,05. RESULTADOS: O grupo exposto ao estresse pelo frio apresentou maior depleção de lipídio nas glândulas adrenais (P<0,05) e de glicogênio no fígado (P<0,05). O grupo induzido à hipotermia mostrou menor volume do núcleo de seus cardiomiócitos (108 + 1,7 µm³; P<0,05), reduziu em 76 por cento comparado ao grupo controle (142 + 2,3 µm³). Correlação de Bartko: CON=0,44; IH=0,96, a variação entre a média dos grupos foi significativamente diferente. CONCLUSÃO: Esses resultados sugerem que a exposição ao estresse agudo pelo frio induz redução do núcleo dos cardiomiócitos em ratos.

INTRODUCTION: Total body induced hypothermia and myocardial cooling are effective methods regarding myocardial protection during heart surgery and ischemia. It is described in previous studies that extreme low temperature exposure causes mitochondrial cristae and myofilament disarrangement in cardiomyocytes, however, no investigation has analyzed the effects of cold stress on nuclear size of cardiomyocytes. OBJECTIVES: To evaluate the effects of acute cold stress exposure on the nuclear size of cardiomyocytes in rats. METHODS: The experimental study procedures were performed on 300-310g adult male Wistar rats. Rats (n=20) were divided into two groups: 1) Control (CON) and; 2) Induced hypothermic (IH) group. Animals of IH group were exposed during 4 hours once at a controlled temperature of - 8ºC. It was performed histological analysis of liver and adrenal gland to examine the stress condition of animals. Cardiomyocytes nucleus size were examined by three independent investigators with the same and standardized criteria and analyzed by Bartko's intra-class correlation coefficient (R>0.75 = positive concordance). Student's t test was applied. The significance level was set at P<0.05. RESULTS: The induced hypothermic group presented higher lipid depletion in adrenal gland cells (P<0.05) and higher glycogen depletion in liver glycogen (P<0.05). The experimental group showed lower cardiomyocytes nuclear volume (108 + 1.7 µm³; P<0.05), it decreased in 76 percent compared to the control group (142 + 2.3 µm³). Bartko's correlation: CON=0.44; IH=0.96, variation analysis between group's means differences was significant. CONCLUSION: These data suggest that acute cold stress exposure induces cardiomyocytes nucleus size reduction in rats.

Correlation of the Nuclear beta-catenin Expression with the Clinicopathological Parameters of Hepatocellular Carcinoma

BACKGROUND: Hepatocellular carcinoma (HCC) is the most common primary malignant tumor of the human liver. However, the molecular changes and mechanisms that regulate the development and progression of HCC remain unclear. Beta-catenin is known as a multi-functional protein that acts as a regulator of the cadherin-mediated cell-cell adhesion system and also in the Wingless/Wnt signal transduction pathway. The aim of this study was to investigate the expression of beta-catenin and its possible role in HCC. METHODS: We investigated the expression of beta-catenin, Ki-67, TP53, alpha-smooth muscle actin and CD34 by performing immunohistochemical staining for 61 specimens of HCC and their adjacent non-tumorous tissue. We also examined the relationship between the nuclear expression of beta-catenin and the clinicopathologic parameters. RESULTS: The altered expression of beta-catenin was not detected in the nontumorous liver tissue. The nuclear expression of beta-catenin was observed in approximately 16% (10/61) of the HCC specimens. Double immunohistochemical staining for beta-catenin and E-cadherin showed a close relationship between nuclear translocation of beta-catenin and the loss of the membranous E-cadherin expression. Significant correlation was found between the nuclear translocation of beta-catenin and the tumor size, tumor necrosis and the presence of microvessel invasion and intrahepatic metastasis (p<0.05). CONCLUSIONS: This data indicates that nuclear translocation of beta-catenin could play a role in the growth and progression of HCC.