Determination of carbohydrate chains in two kinds of microvascular endothelial cells by lectin cytochemistry / 解剖学报

Objective To determine the expression pattern of carbohydrate chains in two kinds of microvascular endothelial cells (MVECs). Methods Rat jejunal mucosal MVECs were thawed, and lung tissues were removed from specific pathogen free piglet of 3 days old to isolate and culture porcine pulmonary MVECs by collagerase digestion and differential attachment. By lectin cytochemistry, staining of 8 lectins including concanavalin A (Con A), phaseolus vulgaris erythroagglutinin (PHA-E), ricinus communis agglutinin I (RCA-I), lycopersicon esculentum lectin (LEL), sambucus nigra lectin (SNA), ulex europaeus agglutinin I (UEA-I), wheat germ agglutinin (WGA) and dolichos biflorus agglutinin (DBA) was detected in rat jejunal mucosal and porcine pulmonary MVECs. Results In rat jejunal mucosal MVECs, strong positve staining was present for Con A, WGA and LEL, medium one for PHA-E N SNA and RCA-I, weak one for DBA, and negative staining for UEA-I. In porcine pulmonary MVECs, strong positive staining was present for Con A and PHA-E, medium one in RCA-I, weak one for LEL and SNA, and negative staining for UEA-I, WGA and DBA. Conclusion The carbohydrate patterns in two kinds of MVECs display significant heterogeneity. Both rat jejunal mucosal and porcine pulmonary MVECs express mannose, galactose, 1, 3-N-acetylglucosamine and sialic acid at different levels. N-acetylglucosamine and N-acetylgalactosamine are detected in the former but not in the latter, and fucose do not in both MVECs.

Preparation of Quality Control Slides and Establishment of an External Quality Assessment Program for Five Special Stains Routinely Used in Diagnostic Hematology

BACKGROUND: In general, internal/external quality control of special stains for diagnosis of hematological diseases may be unavailable in a clinical laboratory owing to the lack of an appropriate positive/negative control material. METHODS: We developed a protocol on positive/negative control materials for five special stains (iron, myeloperoxidase [MPO], periodic acid-Schiff [PAS], Sudan black B [SBB], and alpha-naphthyl acetate esterase [ANAE]) using a hematological malignant cell line. First, we compared stainability of seven cell lines (HL-60, THP-1, K562, Kasumi-1, KG-1, KO52, and NKM-1), then confirmed duration of stable stainability. A proficiency test using external quality control materials was conducted at eleven institutions, which participated voluntarily. RESULTS: HL-60 and THP-1 cell lines, which showed good stainability among the seven cancer cell lines, were selected as external quality control materials. The stainability of a prepared cell line fixed on control slides was stable for 3–4 weeks (MPO, SBB, and PAS) or 9–10 weeks (ANAE). The stainability of paraffin-embedded control material for iron stain was stable for 3 months. The results from 11 institutions were the same on iron, MPO, SBB, and ANAE. Nevertheless, two of 10 institutes showed discrepant results on PAS. CONCLUSIONS: In this study, we demonstrated that cell lines could serve as a standard quality control material for special stains. Most institutions showed representative results on special stains except for PAS. This protocol for special stain may be useful as an external or internal quality control in a haematology laboratory.

Características clínicas y epidemiológicas de los pacientes hospitalizados con derrame pleural / Clinical and epidemiological characteristics of patients hospitalized with pleural effusion

El derrame pleural tiene una prevalencia mundial de aproximadamente 400 de cada 100.000 habitantes y Venezuela tiene cifras similares. Se relaciona con múltiples patologías, por lo que determinar sus características podría ayudar a obtener un mejor diagnóstico y tratamiento. Métodos: Se realizó un estudio de casos, retrospectivo y descriptivo, obteniendo información de las historias de pacientes hospitalizados con derrame pleural en el servicio de Medicina Interna del Hospital Dr. Domingo Luciani en el período Enero 2010- Abril 2015. Resultados: La edad promedio fue de 49±19 años, género masculino (53%). Motivo de consulta: disnea (81%), dolor torácico (44%) y tos (37%). Los síntomas: disnea (92%), dolor pleurítico (58%) y fiebre (54%). Antecedentes personales: HTA (32%), DM (22%) e IC (20%). Radiografía de tórax: (60%) derrame pleural derecho y (26%) izquierdo. Tomografía de tórax realizada en (77%). Citoquímicos: (85%) exudado (53% mononuclear y 32% polimorfonucleares). La prueba de ADA positiva en 25%, cultivo para bacterias realizado en 89 casos, positivos 18%. Bloque celular con resultado inflamatorio (80%). Biopsia pleural realizada (22%): inflamatorio (36,4%), seguido por ADC metástasico (31,8%). Estancia hospitalaria > 15 días (76%) y el diagnóstico final fue infeccioso (51%). Conclusión: Contando con estos datos clínicos- epidemiológicos se puede caracterizar el comportamiento del derrame pleural en nuestro centro para el rápido y acertado diagnóstico, igualmente proponer una investigación prospectiva donde se analice el comportamiento de dicha enfermedad, y crear protocolos de actuación(AU)

Pleural effusion has a worldwide prevalence of approximately 400 per 100,000 inhabitants and Venezuela has similar statistics. It is related to multiple pathologies, which determine their characteristics which could help for better diagnosis and treatment. Methods: A retrospective descriptive case study was conducted, obtaining information from the charts of hospitalized patients with pleural effusion in Internal Medicine Dr. Domingo Luciani Hospital Venezuela in the period January 2010-April 2015. Results: Mean age 49 ± 19 years, male genre (53%). Complaints: dyspnea (81%), chest pain (44%) and cough (37%). Symptoms: dyspnea (92%), pleuritic pain (58%) and fever (54%). Personal history: hypertension (32%), DM (22%) and HF (20%). Chest x-ray: right pleural effusion (60%), left (26%). Chest tomography performed on (77%). Cytochemical: exudate: 85% (53% mononuclear and polymorphonuclear 32%). ADA testing positive in 25%. For bacteria culture: performed in 89 cases, 18% positive. Cell block inflammatory (80%). Pleural followed by metastatic ADC (31.8%). Hospital stay> 15 days (76%) and final diagnosis was infection (51%). Cause of discharge from hospital: improvement (80%). Conclusion: Having these clinical and epidemiological data can characterize the behavior of pleural effusion for quick and accurate diagnosis(AU)

Características morfo-citoquímicas del líquido pleural: estudio de 104 casos / Morph-cytochemical characteristics of the pleural effusion: study of 104 cases

El objetivo del trabajo es analizar las características morfo-citoquímicas de los líquidos pleurales, de pacientes con derrame pleurales, para proporcionar métodos útiles de diagnóstico etiológico. Se extrajeron entre 10 y 50 ml. de líquido pleural de 104 pacientes. Los líquidos pleurales se clasificaron en exudados y trasudados. Se practicó estudio morfo-citoquímico, citología, coloración de Ziehl-Neelsen y cultivo para bacilo de Koch, hongos y biopsia. La distribución de los derrames pleurales fue: tuberculosos (n=36-35,0%); neoplásicos (n=28-27,0%); indeterminados (n=18-17,3%) infecciones respiratorias (11,0%); insuficiencia renal (7,0%) y otros. El 84,0% de los derrames pleurales fue de tipo exudado. El 50,0% de los líquidos pleurales hemorrágicos fueron neoplásicos. En los derrames pleurales de tipo exudado hubo una predominancia de linfocitos/polimorfonucleares. La coloración de Ziehl-Neelsen resultó siempre negativa. Los cultivos fueron positivos para bacilo de koch (10,0%). En el 17,9% se diagnosticaron derrames pleurales neoplasicos. Concluimos, que las características morfo-citoquímicas del líquido pleural son importantes para hacer un diagnóstico etiológico del derrame pleural aun cuando se disponga de métodos más actualizados

The objective of the study is to analyze the characteristics of pleural fluid patients with pleural effusions to provide useful methods of etiological diagnosis. It was extracted between 10 and 50 ml pleural fluid 104 patients. The pleural fluids were classified in transudates and exudates. Was precticed morph-cytochemical, cytology, staib of Ziehl-Neelsen and cultivation for Koch's bacilli and fungi and biopsy. The distribution of the pleural effusions was: tuberculosis (n=36-35.0%); neoplastic (n=28-27.0%), indeterminate (n=18-7.3%) respiratory infections (n=18-17.3%); kidney failure (7,0%) and others. The 84.0% of the pleural fluids was exudates type. The 50.0% of bleeding pleural fluids were neoplastic. There was a predominance of lymphocytes/polymorphonuclears in the pleural effusions of exudates type; the stain of Ziehl-Neelsen was always negative. The cultures were positive for Koch's bacilli (10.0%). Neoplastic effusions were diagnosed at 17.0%. We conclude that the morph-cytochemical features of the pleural fluid are important making a diagnosis etiological effusion even when most up-to-date methods are available

Ultrastructure and blood-iris barrier in experimental rubeosis iridis in rabbit

Iris neovascularization was produced in rabbits by hypotony following repeated aspiration of the vitreous. The hypotony was produced after 0.3 ml of vitreous fluid was aspirated using a 25-gauge needle through the pars plana of 10 rabbits. For the histochemical study, horseradish peroxidase(HRP) was injected through the ear lobe vein. After fixation of the iris tissue, the tissue was treated with diaminobenzidine and examined with both light microscopy and transmission electron microscopy. The newly-formed vessel was abundant, particularly on the upper stroma of the iris. The new vessel formation was evident due to the proliferation of endothelial cells, which may have been derived from preexisting iris vessels. The endothelial cells of the newly-formed vessels revealed prominent villous processes into the vascular lumen, formation of the marginal flap, numerous fenestrations in the endothelial junction, and reaction product onto extravascular space by the cytochemical electron microscopy. These results suggest that hypotony in the rabbit produces the disruption of the blood-iris barrier and the balance between angiogenesis-antiangiogenesis modulation.