Histomorphometric analysis of liver biopsies of treated patients with Gaucher disease type 1

Gaucher disease (GD) is an autosomal recessive lysosomal disorder caused by a disturbance in the metabolism of glucocerebroside in the macrophages. Most of its manifestations - hepatosplenomegaly, anemia, thrombocytopenia, and bone pain - are amenable to a macrophage-target therapy such as enzyme replacement. However, there is increasing evidence that abnormalities of the liver persist despite the specific GD treatment. In this work, we adapted histomorphometry techniques to the study of hepatocytes in GD using liver tissue of treated patients, developing the first morphometrical method for canalicular quantification in immunohistochemistry-stained liver biopsies, and exploring histomorphometric characteristics of GD. This is the first histomorphometric technique developed for canalicular analysis on histological liver biopsy samples.

Application of DNA-image cytometry in diagnosis of breast cancer / 中华内分泌外科杂志

Objective To determine the value of DNA image cytometry (DNA-ICM) in diagnosis of breast cancer.Methods Totally 187 patients with breast or axillary mass admitted from Aug.2014 to Oct.2017 were included in the study.Fine-needle aspiration cytology (FNAC),DNA-ICM and liquid-based cytology (LBC) were employed.The results showed that 100 cases were breast cancer patients and 73 cases were benign patients.DNA-ICM and LBC were used to compare the sensitivity and specificity of the two methods.Results The sensitivity and specificity were 82.0％ and 100％ for DNA-ICM,and 69.0％ and 97.3％ for LBC.The sensitivity of DNA-ICM was significantly higher than that of LBC (P=0.033).In breast tissue,the sensitivity and specificity of DNA-ICM were 83.3％ and 100％,and they were 64.8％ and 97.5％ for LBC.The difference of sensitivity was statistically significant (P=0.028).In axillary tissue,the sensitivity and specificity of DNA-ICM were 80.4％ and 100％,and they were 73.9％ and 97.0％ for LBC.The difference was not statistically significant (P=0.456).The sensitivity of joint detection was 93.5％,significantly higher than that of LBC alone (P=0.024).The effect of joint detection was better than that of LBC.Conclusion DNA-ICM method is of great value in the diagnosis of breast cancer,which can be combined with LBC in diagnosis of breast cancer.

Effects of aldehyde-free fixation solution and aldehyde fixation solution on the nucleus DNA and cytoplasm staining / 重庆医学

Objective To investigate the effect of aldehyde-free fixation solution and aldehyde fixation solution on DNA and cytoplasm of nucleus.Methods The DNA quantification (IOD),discrete coefficient (CV),DNA index (DI) and nucleus area (area) were measured by DNA quantification system (ICM) in 8 different fixation solution;combined with cytological staining,the effects of different fixative solution on nuclear DNA and cytoplasmic staining were analyzed.Results Aldehyde fixation solution was better than aldehyde free fixation solution,the formaldehyde in fixtion solution has important influence on Feulgen-eosin staining.Conclusion Aldehyde fixation solution combined with cytological staining can obtained good dyeing effect,which provides the basis for the subsequent multi-technique joint diagnosis.

Identification of Epstein-Barr Virus in the Human Placenta and Its Pathologic Characteristics

Epstein-Barr virus (EBV), a common pathogen in humans, is suspected as the cause of multiple pregnancy-related pathologies including depression, preeclampsia, and stillbirth. Moreover, transmission of EBV through the placenta has been reported. However, the focus of EBV infection within the placenta has remained unknown to date. In this study, we proved the expression of latent EBV genes in the endometrial glandular epithelial cells of the placenta and investigated the cytological characteristics of these cells. Sixty-eight placentas were obtained from pregnant women. Tissue microarray was constructed. EBV latent genes including EBV-encoding RNA-1 (EBER1), Epstein-Barr virus nuclear antigen 1 (EBNA1), late membrane antigen (LMP1), and RPMS1 were detected with silver in situ hybridization and/or mRNA in situ hybridization. Nuclear features of EBV-positive cells in EBV-infected placenta were compared with those of EBV-negative cells via image analysis. Sixteen placentas (23.5%) showed positive expression of all 4 EBV latent genes; only the glandular epithelial cells of the decidua showed EBV gene expression. EBV infection status was not significantly correlated with maternal, fetal, or placental factors. The nuclei of EBV-positive cells were significantly larger, longer, and round-shaped than those of EBV-negative cells regardless of EBV-infection status of the placenta. For the first time, evidence of EBV gene expression has been shown in placental tissues. Furthermore, we have characterized its cytological features, allowing screening of EBV infection through microscopic examination.

Application of DNA-image cytometry in the diagnose of urothelial cell carcinomas / 实用医学杂志

Objective To investigate the value of DNA-image cyt-ometry (DNA-ICM) in the diagnosis of urothelial cell carcinomas (UCC). Methods Totally 162 voided urine specimens (92 cases from urothel-ial car-cinomas patients and 70 cases from benign urinary system diseases patients ) were detected with DNA-ICM and liquid-based cytology (LBC), respectively. Results The sensitivity and specificity of DNA-ICM were 65.2%and 100% respectively in the diagnosis of UCC but those of LBC were 27.2% and 98.6%, respectively. The sensitivity of DNA-ICM was significantly higher than that of LBC in the diagnosis of UCC (P 0.01). Conclusion DNA-ICM, which improves the positive rate of urinary cytology, has great application value in the diagnosis of urothelial cell carcinomas and it is an effective screening method for urothelial cancer in diag-nosis and follow-up.

The combination of flow cytometry and automated cell image analyzer in determining the nature of ascites and pleural effusion nature / 中华检验医学杂志

Objective To explore the clinical value of flow cytometry( FCM) and DNA automated cell image analyzer ( AICM) in determine the character of ascites and pleural effusion.Methods This was a cross-sectional study.203 ascites and pleural effusionsamples were random selected from PLA hospital inpatients between August 2013 to June 2014 .The DNA content of sediment cells were detectedthrough the FCM and AICM respectively benign and malignant disease were differentiated according the counts and proportion of aneuploid cells.The sensitivity, specificitywere calculated byROC curves.Results The sensitivity, specificity and accuracy of flow cytometry cell in detectingtumor cells were 78.6%,80.0% and 79.2%%, while the sensitivity, specificity and accuracy of image analyzer were 83.5%,78.6% and 81. 3%respectively.When FCM and AICMwere combined ,the sensitivity, specificity and accuracyincreased to 92.2%, 86.3% and 89.6%.Conclusions Compared toconventional cytology test, the sensitivity and specificity were significantly high when the two methods were combined .Therefore, the combination method can be used to assist in clinical identification of the nature of ascites and pleural effusion and to help the diagnosis of disease.

Application of DNA image cytometry in distinguishing benign and malignant pleuroperitoneal fluids / 安徽医科大学学报

Objective To investigate the value of DNA image cytometry in thediagnose of benign and malignant pleuroperitoneal fluids by comparing with the liquid-based cytological results. Methods There were 417 cases in-volved in this study. Pap stain for cytology analysis and feulgen stain for DNA image cytometry were used to identify benign and malignant pleuroperitoneal fluids respectively,then compared the results of the two methods. Results Clinically or pathologically,213 were classified as malignant and the other 204 as benign. The sensitivity, specifici-ty,accuracy, positive predictive value and negative predictivevalue of DNA image cytometry were 89. 7%,100%, 94. 7%,100%,and 90. 3%,respectively. However,the sensitivity,specificity,accuracy, positive predictive value and negative predictive value of liquid-based cytology were 63. 4%, 81. 9%, 72. 4%, 78. 5%, and 68. 2%, re-spectively. There were significant differences in thesensitivity, specificity,accuracy, positive predictive value and negative predictivevalue. Conclusion DNA image cytometry has great application value in the diagnosis of benign and malignant pleuroperitoneal fluids, and can increase the positive rate, reduce misdiagnosed rate with liquid-based cytology.

THE VALUE OF CELL DNA QUANTITATIVE ANALYSIS IN THE EARLY SCREENING OF CERVICAL CANCER AND PRE-CANCERATION / 现代医院

Objective To evaluate the application of DNA image cytometry in screening cervical intraepithe-lial neoplasia and cervical cancer.Methods 80 patients with cervical intraepithelial neoplasia admitted during No-vember 2012 and November 2014 were involved in this study.The samples from all the patients were taken by cervix brushes and prepared for 2 slides:one for liquid-based cytology staining, read by cytologist, and the other for Thi-onin-Feulgen staining, tested by DNA image cytometry.The biopsy was performed with direction of colposcopy if one test was positive to verify the sensitivity and specificity of the two methods of staining.Results 70 cases were positive by DNA image cytometry and 27 ones were positive by liquid-based cytology.There was statistically signifi-cant difference in the sensitivity between the DNA image cytometry and the liquid -based cytology ( 94.8% vs. 38.8%;p<0.05), but no difference in the specificity between them (99.6%vs.99.8%).Conclusion Besides its good specificity, DNA image cytometry is more sensitive in screening cervical intraepithelial neoplasia and cervical cancer.

The prostate after administration of anabolic androgenic steroids: a morphometrical study in rats

Purpose Many adverse effects have been associated with abuse of anabolic-androgenic steroids (AAS), including disorders of the urogenital tract. The objective of this study is to analyze the morphological modifications in the prostate ventral lobe of pubertal and adult rats chronically treated with AAS, using morphometric methods. Materials and Methods: We studied 39 male Wistar rats weighing between 400 g and 550 g. The rats were divided into four groups: (a) control rats, with 105 days of age (C105) (n = 7); (b) control rats with 65 days of age (C65) (n = 9), injected only with the vehicle (peanut oil); (c) treated rats, with 105 days of age (T105) (n = 10) and (d) treated rats with 65 days of age (T65) (n = 13). The treated rats were injected with nandrolone decanoate at a dose of 10 mg.Kg-1 body weight. The steroid hormone and the vehicle were administered by intramuscular injection once a week for eight weeks. The rats were killed at 161 days of age (C105 and T105) and 121 days of age (C65 and T65) and the ventral prostate lobe was dissected and processed for histology. The height of the acinar epithelium, the surface densities of the lumen, epithelium and stroma were observed with X400 magnification using an Olympus light microscope coupled to a Sony CCD video camera, and the images transferred to a Sony monitor KX14-CP1. The selected histological areas were then quantified using the M42 test-grid system on the digitized fields. The data were analyzed with the Graphpad software. To compare the quantitative data in both groups (controls and treated) and the outcomes, Student's t-test was used (p < 0.05 was considered significant). Results: The weight (p < 0.001) and volume (p = 0.004) of the prostate ventral lobe showed differences between C65 and T65 groups and between C105 and T105 groups. The epithelium height showed no difference between groups C65 and T65 (p = 0.8509), but the T105 group showed an increase of 32% compared ...

Techniques of digital image analysis for histological quantification of melanin / Técnicas de análise de imagem digital para quantificação histológica da melanina

Morphometric analysis of tissue melanin may quantitatively contribute to research on pigmentation disorders. The authors present three methods for image analysis, which allow for identification of melanin-equivalent pixels in the epidermis using Fontana-Masson stain and, therefore, for the calculation of its percentage in the different epidermal layers. Moreover, they discuss the main elements related to the analysis and the need for rigorous standardization of the process.

A análise morfométrica da melanina tecidual pode subsidiar quantitativamente a pesquisa em discromias. Os autores demonstram três técnicas de análise de imagem digital que permitem a identificação dos pixels equivalentes à melanina na epiderme pela coloração de Fontana-Masson, possibilitando o cálculo da sua porcentagem nas diferentes camadas da epiderme, e discutem os principais elementos relacionados à análise e a necessidade de rigorosa padronização do processo.

DNA image cytometry in breast fine needle aspiration cytology / 肿瘤研究与临床

Objective To discuss the diagnosis value of DNA imaging cytometry (DNA-ICM) analysis in fine needle aspiration cytology (FNAC).Methods Conventional cytopathology and DNA-ICM were used to detect 95 fine needle aspiration patients with breast masses.The results were analyzed according to the histopathological diagnosis.Results There was not significant difference between conventional cytopathology and DNA-ICM [88.4 ％(84/95) vs.94.7 ％(90/95)] (x2 =2.457,P =0.117).Two false positive cases had been found in conventional cytopathologic diagnosis,but not in DNA-ICM.There was a positive correlation between the nucleus area and its DNA content (r =0.744,P =0.000).By DNA-ICM,the detectable rate of abnormal DNA content in the infiltrating ductal carcinoma was higher than that in the infiltrating lobular carcinoma [90.3 ％(65/72) vs.50.0 ％(4/8)] (P =0.011).Conclusion The detection of DNA-ICM can reduce the false positive rate of FNAC,but it has limitations in diploid tumors.

Análise morfométrica do colágeno dérmico a partir da segmentação por conglomerados (clusters) de cor / Morphometric analysis of dermal collagen by color clusters segmentation

Análise morfométrica do colágeno dérmico pode fornecer subsídio quantitativo para a pesquisa em dermatologia. Os autores demonstram uma técnica de análise de imagem digital que permite a identificação de estruturas microscópicas, a partir da segmentação por conglomerados (clusters), de cor aplicada à estimativa da intensidade e densidade das fibras colágenas da derme.

Morphometric analysis of dermal collagen can provide quantitative support to dermatologic research. The authors of this article disclose a technique of digital image analysis which allows the identification of microscopic structures by color cluster segmentation regarding the estimate intensity and density of dermal collagen fibers.

Estudo citofotométrico da expressão dos marcadores tumorais Ki-67 e CD34 no adenocarcinoma de próstata / Cytophotometric expression of tumor antigen markers Ki-67 and CD-34 in prostate adenocarcinoma

OBJETIVO: Quantificar a porcentagem da imunomarcação no índice de marcagem e densidade óptica do Ki-67 e CD34 no adenocarcinoma de próstata e compará-las entre si. MÉTODOS: Foram estudados, através de imunoistoquímica, o Ki-67 e o CD34 em 34 casos de adenocarcinoma de próstata provenientes de prostatectomia radical no período de 2000 a 2005 realizado no Hospital Regional do Gama em Brasília. Estes marcadores foram quantificados através do software SAMBA 4000 ® Sistema de Análise Microscópica de Busca Automática e do software IMMUNO® para análise das variáveis índice de marcagem e densidade óptica. Para avaliação da associação entre as expressões do marcador, foi estimado o coeficiente de correlação de Spearman. Para a comparação do tipo de lesão, foi usado o teste t de Student em amostras pareadas e não paramétrico de Wilcoxon. RESULTADOS: Dos 34 blocos que foram para leitura dos marcadores tumorais, 15 marcaram expressão com Ki-67, 34 com CD34 e 14 com ambos os marcadores. O índice de marcagem do CD34 teve valor mediano de 72,72 por cento, valor mínimo 5,14 por cento e valor máximo 88,81 por cento. O índice de marcagem do Ki-67 teve mediana de 73,78 por cento, mínimo de 16,87 por cento e máximo de 87,47 por cento. A densidade óptica do CD34 teve mediana de 48,33, mínimo de 35,65 e máximo de 85,86. Na densidade óptica do Ki-67 o valor da mediana foi 40,03 sendo a mínima de 21,53 e a máxima de 52,43. CONCLUSÃO: A expressão citofotométrica do Ki-67 teve índice médio de marcação de 64,04 por cento e o CD34 de 61,64 por cento. A expressão citofotométrica da densidade óptica média do Ki-67 foi de 39,49 e no CD34 de 53,69. Há diferença significativa entre a imunomarcação do Ki-67 e CD34 em relação à densidade óptica (p=0,025), não havendo diferença significativa no índice de marcagem (p=0,470).

OBJECTIVE: To quantify the percentage of immunostaining through the labeling index as well as the optical density of Ki-67 and CD34 in prostate adenocarcinoma and compare the results between markers. METHODS: Markers Ki-67 and CD34 were studied using immunohistochemistry in 34 cases of prostate adenocarcinoma from radical prostatectomies performed at the Hospital Regional do Gama in Brasilia, Brazil from 2000 through 2005. Those markers were quantified using the SAMBA 4000® software - Automated Scanning Microscopic Analysis System - and the IMMUNO® software in the analysis of the variables labeling index and optical density. Spearman's correlation coefficient was estimated in order to evaluate the association between the expression levels of the markers. For the comparison of lesion types, Student's paired t-test and the nonparametric Wilcoxon test were used. RESULTS: Of the 34 blocks referred for the study of the tumor markers, 15 were positive for Ki-67, 34 showed CD34 expression, and 14 were positive for both markers. The median value for the labeling index of CD34 was 72.72 percent; the minimum was 5.14 percent and the maximum, 88.81 percent. The median for the Ki-67 labeling index was 73.78 percent, while the minimum was 16.87 percent, and the maximum, 87.47 percent. The median value for the optical density of CD34 was 48.33, the minimum was 35.65 and the maximum, 85.86. For the optical density of Ki-67, the median was 40.03, while the minimum and maximum values were 21.53 and 52.43, respectively. CONCLUSION: The cytophotometric expression of Ki-67 had a mean labeling index of 64.04 percent, and the mean CD34 labeling index was 61.64 percent. The cytophotometric expression of the mean optical density of Ki-67 was 39.49, while for CD34 it was 53.69. There was a significant difference between Ki-67 and CD34 immunostaining with respect to optical density (P=0.025); no significant difference occurred regarding labeling index (P=0.470).

Expressão citofotométrica quantitativa da CASPASE-3 e do CD-34 no carcinoma ductal infiltrante de mama / Cytophotometric expressions of CASPASE-3 and CD-34 in breast cancer

OBJETIVO: Descrever, correlacionar e comparar a expressão dos marcadores tumorais CD-34 (angiogênese) e caspase-3 (apoptose) em carcinoma ductal invasor de mama. MÉTODOS: Foram utilizados 22 casos de adenocarcinoma infiltrante de mama provenientes de blocos de parafina e, após preparo específico para imunoistoquímica, 15 apresentaram leitura satisfatória e foram avaliados pelo sistema de fotocitometria de imagem SAMBA 4000® e software IMMUNO®. Os parâmetros analisados foram o índice de marcagem e densidade óptica. RESULTADOS: Para o CD-34 não houve normalidade dos dados na análise do índice de marcagem, com obtenção de P=0,019, havendo normalidade para a análise da densidade óptica, com P=0,199. Para a caspase-3 houve normalidade de dados para o índice marcagem com P=0,306 e para a densidade óptica com P=0,114; não houve diferença estatística significativa entre eles em relação à média do índice de marcagem (P=0,872) e da densidade óptica (P=0,816), quando analisados os parâmetros que definem a expressão dos marcadores; existiu tendência à associação entre a densidade óptica e o índice de marcagem do marcador tumoral caspase-3, com P=0,025. Não foi observada tendência à associação quando comparados densidade óptica e índice de marcagem do marcador tumoral CD-34; índice de marcagem do marcador tumoral caspase-3 e índice de marcagem do marcador tumoral CD-34; e densidade óptica da caspase-3 com a do CD-34. CONCLUSÃO: Dos 22 casos incluídos foi possível verificar a expressão do marcador CD-34 em 18 lâminas e da caspase-3 em 22 lâminas; Para o CD-34 não houve normalidade dos dados na análise do índice de marcagem, havendo sim normalidade para a análise da densidade óptica. Para a caspase-3 houve normalidade de dados tanto para o índice de marcagem como para a densidade óptica. Existe tendência à associação entre a densidade óptica e o índice de marcagem da caspase-3. Não foi observada tendência quando comparados densidade óptica e índice...

OBJECTIVE: Describe, correlate and compare the expression of the tumor markers CD 34 (angiogenesis) and caspase-3 (apoptosis) in invasive breast adenocarcinoma, through image cytometry with the system SAMBA4000®. METHODS: Twenty-two cases of invasive breast adenocarcinoma from paraffin-embedded archival tissue, and after specific prepare, fifteen cases presented a satisfactory lecture with SAMBA4000® and could, finally, be evaluated by the software IMMUNO® (n = 15). The parameters analysed were the label index - in percentage, indicating the marked surface - and the optical density, in pixels - indicator of the marker intensity. The results were tabulated and expressed in averages, mediums, minimum and maximum values. The statistic analysis was realized by the Shapiro-Wilkins, Student test, Pearson's and Spearman's correlation, with statistic significance for values from p < 0,05. RESULTS: There was no data normality for the label index CD34 (p= 0,019), there was normality in the analysis of the optical density of both markers and label index of the marker Caspase-3. There was no difference relating to the average of the index marker and the optical density when they were compared. CONCLUSIONS: There was a tendency to correlate the label index and the optical density of the tumor marker caspase-3, the same did not occur with the tumor marker CD 34. Other analysis did not show any correlation between the two studied markers. Other studies involving theses two cell processes are needed to extend the knowledge of the cancer biomolecular mechanic and to permit new diagnostic and therapeutic strategies.

Significado clínico-patológico das expressões citofotométricas do Ki-67 e Caspase-3 no carcinoma de células escamosas do esôfago / Clinicopathologic significance of the Ki-67 and Caspase-3 cytophotometric expressions in the esophageal squamous cell carcinomal

RACIONAL: A escolha da forma de tratamento do carcinoma de células escamosa de esôfago ainda hoje é orientada pelo estadiamento tumoral, onde as características histopatológicas do tumor são o maior determinante. Parale-lamente, desenvolvem-se estudos para entender o comportamento da biologia tumoral por método imunoistoquímico de quantificação manual, avaliando a ati-vidade proliferativa ou apoptótica do tecido em análise. As desvantagens conti-das no modo manual fizeram surgir e desenvolver método computadorizado de análise de imagem. OBJETIVOS: Verificar as expressões dos marcadores KI-67 e Caspase-3 e correlacioná-las com as características clínico-patológicas do tumor. MÉTODOS: Foram estudados 29 blocos parafinados provenientes de pa-cientes portadores de carcinoma de células escamosas de esôfago submetidos à esofagectomia e pertencentes a acervos de laboratórios de patologia. Proce-deu-se preparo das lâminas por técnica imunoistoquímica convencional. A quantificação da imunorreatividade às proteínas Ki-67 e Caspase-3 foi realizada pelo software de análise de imagem computadorizada SAMBA (Systeme d'Analyse Microphotometrique a Balayage Automatique) através do índice de marcagem encontrado. RESULTADOS: Predominaram na amostra o sexo mascu-lino (82,7 por cento); maiores de 50 anos; tumores moderadamente diferenciados (68,98 por cento); estágio III (72,42 por cento); lesões >3cm e localizadas no ⅓ inferior do ór-gão. Os índices médios de marcagem identificados foram de 62,05 por cento para o Ki-67 e 86,06 por cento para a Caspase-3, e não mostraram correlação com as caracterís-ticas clínico-patológicas como sexo, idade, estadiamento tumoral, grau de pro-fundidade da lesão e comprometimento linfonodal. Houve significante diferença de expressão do Ki-67 entre os graus histológicos (P=0,047) e correlação entre os índices dos marcadores estudados (r=0,41 e P =0,032). CONCLUSÃO: Na presente investigação as atividades das proteínas estudadas se mostraram in-tensas sendo que a da Caspase-3 foi superior ao Ki-67 mas sem correlação com as características clínico-patológicas.

BACKGROUND: The esophageal squamous cell carcinoma treatment strategy is still based on the tumor staging, where tumor histopathologic charac-teristics are the major determinants. In parallel, studies have been developed in order to better understand the tumor biology using immunohistochemical meth-ods with manual quantification evaluating the proliferative and apoptotic activi-ties of the cells. The disadvantages related to the manual method rose the de-velopment of computerized ways to do the image analysis. OBJETIVES: To verify the expressions of the markers Ki-67 (proliferative) and Caspase-3 (apoptotic) and to correlate them with the clinic and pathologic characteristics of the tumor. METHODS: Twenty-nine paraffin embedded blocks were studied, each one con-taining tissue samples from patients with esophageal squamous cell carcinoma submitted to esophagectomies. The clinic and pathological data were obtained from histopathologic informations and from medical records. The slides were prepared following the routine immunohistochemical method until the point to utilize the specific antibodies (MIB-1 and CPP32). Positive quantification of the immunoreactivity to the proteins Ki-67 and Caspase-3 was performed by the software for computerized image analysis SAMBA (Systeme d' Analyse Micro-photometrique a Balayage Automatique). Statistical analysis was done having P<0.05 considered significant. RESULTS: There was predominance of males (82.7 percent); age over 50 years; moderately differentiated tumors (68.98 percent); tumor stage III (72.42 percent); diameter of lesions >3cm; and lesions located in the lower third of the organ. The mean score indexes found were 62.05 percent for Ki-67 and 86.06 percent for Caspase-3 and there was no correlation with the clinic or pathologi-cal characteristics as gender, age and tumor staging. There was significant dif-ference of Ki-67 expression among the histological grades (P=0.047) and corre-lation between the evaluated indexes (r=0.41 and P=0.032). CONCLUSION: The protein expressions were high and the Caspase-3 protein activity was higher than the Ki-67, without correlation with clinic or pathological characteristics as gender, age, tumor staging, grade of lesion depth and lymph node invasion.

Expressão dos marcadores de angiogênese fator VIII e CD-34 no adenocarcinoma gástrico / Expression of angiogenesis markers factor VIII and CD-34 in gastric adenocarcinoma

RACIONAL: O câncer gástrico continua sendo objeto de grande número de estudos que tentam entender melhor sua gênese, e conseqüentemente propor novos tratamentos. Na atualidade destaque especial está sendo dado na marcação imunoistoquímica onde estão sendo utilizados marcadores diversos sem se saber ainda qual ou quais são os mais efetivos no diagnóstico. OBJETIVO: Identificar e quantificar por citometria de imagem a expressão dos marcadores de angiogênese Fator VIII e CD-34 em tumores gástricos. MÉTODOS:Foram utilizados 29 casos de adenocarcinomas gástricos, todos oriundos de material arquivado e conservado em blocos de parafina. Após a desparafinização, realizou-se coloração com marcadores imunoistoquímicos de angiogênese Fator VIII e CD-34, e as lâminas coradas foram submetidas a estudo citofotométrico de imagem em sistema informatizado SAMBA 4000. RESULTADOS: Foram comparados os dois parâmetros oferecidos pelo método, índice de marcagem e densidade óptica, apenas nos 17 casos em que ocorreu identificação de ambos marcadores. Observou-se expressão numericamente significativa de ambos, porém o Fator VIII apresentou melhor média de densidade óptica, enquanto o CD-34 melhor resultado quanto ao índice de marcagem. CONCLUSÃO:A citometria de imagem foi capaz de identificar e quantificar a expressão do Fator VIII e CD-34 de maneira confiável e satisfatória demonstrando presença de angiogênese, sendo que o Fator VIII marcou menores áreas com melhor qualidade e o CD-34 marcou maiores áreas com menor qualidade.

BACKGROUND: Gastric cancer remains a subject of great interest in several studies searching for understanding its genesis and, therefore, proposing new treatments. Currently special credits have been given to immunohistochemistry as various markers are being tested with uncertain efficacy on diagnosis. AIM: To identify and quantify the expression of Factor VIII and CD-34, angiogenesis markers, in gastric tumors by imaging cytometry. METHODS: Twenty-nine 29 gastric adenocarcinomas sampled-tissue in paraffin blocks underwent immunohistochemical angiogenesis markers evaluation for factor VIII and CD-34 and the marked slides were submitted to SAMBA 4000 reading. RESULTS:The investigation method offered two comparison parameters, marking index and optical density, showing up both in 17 cases. Significant numerical expression had been observed for both markers, but Factor VIII showed a better optical density average while CD-34 presented a better result for marking index. CONCLUSIONS: Image cytometry identified and quantified Factor VIII and CD-34 expressions in a reliable and satisfactory manner, revealing the presence of angiogenesis. Factor VIII enhanced in smaller areas with better quality while CD-34 enhanced greater areas with lower quality.

Prognostic value of DNA ploidy and nuclear morphometry in renal cell carcinoma / 重庆医科大学学报

Objective:To determine the prognostic value of DNA ploidy and nuclear morphometry,and its relations to nuclear grading in renal cell carcinoma.Methods:Pathological grading were done in 47cases of renal cell carcinoma in accordance with the system proposed by Fuhrman et al.Feulgen staining were performed in the tumor histological slides,and the technique of image cytometry was used to quantitate nuclear DNA ploidy and nuclear morphological parameters.The results combined with the data of clinical follow up were analyzed.Results:The mean DNA ploidy in grade Ⅰ～Ⅱ was mainly 2C,≥5C cells less than 10%;the 2C cells in grade Ⅲ～Ⅳ were lower than in grade Ⅰ～Ⅱ,≥5C cells were more than 20%.Among the four grades DNA ploidy were statistically different( P

The significance of DNA content in oral squamous cell carcinoma / 实用口腔医学杂志

Objective: To study the significance of DNA content in oral squamous cell carcinoma (SCC).Methods: DNA content and cell cycle of the cells in 18 cases of oral SCC were analized by image cytometry (ICM). Results: The DNA index (DI) and proliferation index (PI) were remarkably heigher in SCC than in normal epithelium ( P

VALUE OF IMAGE CYTOMETRY ANALYSIS IN DIAGNOSIS OF BRONCHOPULMONARY BENIGN AND MALIGNANT CHANGES / 肿瘤研究与临床

Objective: To study the DNA ploidy and DNA index (DI) in human benign and malignant bronchopulmonary change, and evaluate their value in diagnosis of benign and malignant bronchopulmonary diseases.Methods: The DNA ploidy and DI were analysed by image cytometry analysis (ICM) in 39 cases of bronchitis, 45 cases of squamous metaplasia, 45 cases of dysplasia,and 63 cases of carcinoma.Results: The mean DI for bronchitis and squamous metaplasia were 0.92 and 1.03 respectively, and dysplasia was 1.34,while the mean value for carcinoma was 1.72.Among the four groups,the difference was of evident statistical significance.Conclusion: The DNA content and DNA aneuploidy are with different percentage in benign and malignant bronchopulmonary pathological change.Quantitative analysis of the DNA ploidy and DI may be beneficial in diagnosis and differencial diagnosis of benign and malignant changes,which can provide objective data for clinical treatment and prognosis.

A Comparative Study of DNA Quantitation by Image Cytometry and Flow Cytometry

There are substantial evidences suggesting that DNA content of tumors may provide the prognostic information with independent significances. In this study, the results of DNA ploidy analyzed by image cytometry on touch imprint and by flow cytometry on fresh cell suspension of 78 solid tumors were compared. For 68 cases, there was an excellent correlation between two methods. For 6 cases, an aneuploid population was found by image, but not by flow cytometry one case had an aneuploid peak by flow cytometry. Two methods may use in a complementary fashion m identify as many aneuploid cell population as possible.