Ethanol extract of Kalopanax septemlobus leaf inhibits HepG2 human hepatocellular carcinoma cell proliferation via inducing cell cycle arrest at G1 phase

OBJECTIVE@#To investigate the effects of an ethanol extract of Kalopanax septemlobus (Thunb.) Koidz. leaf (EEKS) on cell proliferation in human hepatocellular carcinoma cells and its mechanisms of action.@*METHODS@#Cells were treated with EEKS and subsequently analyzed for cell proliferation and flow cytometry analysis. Expressions of cell cycle regulators were determined by reverse transcriptase polymerase chain reaction analysis and Western blotting, and activation of cyclin-associated kinases studied using kinase assays.@*RESULTS@#The EEKS suppressed cell proliferation in both HepG2 and Hep3B cells, but showed a more sensitive anti-proliferative activity in HepG2 cells. Flow cytometry analysis revealed an association between the growth inhibitory effect of EEKS and with G1 phase cell cycle arrest in HepG2 cells, along with the dephosphorylation of retinoblastoma protein (pRB) and enhanced binding of pRB with the E2F transcription factor family proteins. Treatment with EEKS also increased the expression of cyclin-dependent kinase (CDK) inhibitors, such as p21WAF1/CIP1 and p27KIP1, without any noticeable changes in G1 cyclins and CDKs (except for a slight decrease in CDK4). Treatment of HepG2 cells with EEKS also increased the binding of p21 and p27 with CDK4 and CDK6, which was paralleled by a marked decrease in the cyclin D- and cyclin E-associated kinase activities.@*CONCLUSIONS@#Overall, our findings suggest that EEKS may be an effective treatment for liver cancer through suppression of cancer cell proliferation via G1 cell cycle arrest. Further studies are required to identify the active compounds in EEKS.

Avaliação comparativa da sedimentoscopia urinária realizada pelo nefrologista e pelo profissional de análises clínicas em pacientes com glomerulopatias / Comparative assessment of urinary sediment done by the nephrologist and by the clinical analyst professional in patients with glomerulonephritis

Introdução: A sedimentoscopia urinária com microscópio munido com contraste de fase (MCF) deveria ser a primeira etapa na determinação da origemdas hematúrias. Objetivo: Avaliar discrepâncias nas descrições dos parâmetros urinários relacionados à origem das hematúrias, comparando as descrições do nefrologista (Nef) e do profissional de análises clínicas (PAC). Métodos: Urinas de pacientes com glomerulopatias (GP) confirmadas por biópsia renal foram analisadas sob MCF, por um Nef e um PAC, ambos sem conhecimento prévio da origem das amostras. Cilindros hemáticos, acantocitúria ou células G1 >5% e dismorfismo eritrocitário foram utilizados na localização glomerular das hematúrias. Resultados: Dos 28 pacientes, 13 pacientes (46,4%) apresentavam glomerulonefrites não proliferativas e 15 (53,6%) glomerulonefrites proliferativas. Comparativamente ao PAC, o Nef identificou maior número de hemácias (mediana/mL de urina, 80.000 vs 4.800, p=0,001), maior número de cilindros hemáticos (39,3% vs 0%, p=0,001), maior freqüência de acantocitúria ou células G1 >5% (35,7% vs. 7,14%, p=0,021) e de dismorfismo eritrocitário (96,2% vs 7,14%, p<0,001). As discrepâncias dos resultados permaneceram após a separação das glomerulopatias em proliferativas e não proliferativas. Conclusão: Os parâmetros urinários que caracterizam a origem da hematúria foram mais freqüentemente identificados pelo nefrologista e sugerem que a urinálise, pela sua simplicidade e grande valor informativo, deveria ser incluída obrigatoriamente nos programas de treinamento em nefrologia.

Introduction: In the assessment of hematuria, the first step should be the identification of the origin of the bleeding, which can be done easily by analyzing the urine under phase-contrast microscopy. Obective: To assess the discrepancy of reports of the urinary parameters utilized in the localization of the glomerular origin of hematuria, comparing reports by the nephrologists and by the clinical laboratory technologist. Methods: Urines of patients with biopsy proven glomerulonephritis were assessed under phase-contrast microscopy by a nephrologist and a clinical laboratory technologist, both without previous knowledge of the origin of the samples. Red blood cell (RBC) casts, urinary acanthocytes or G1 cells >5%, and erithrocyte dysmorphism were used tolocalize the glomerular bleeding. Results: Among 28 patients, 13 (46.4%) had non proliferative glomerulonephritis and 15 (53.6%) had proliferative glomerulonephritis. Relatively to the clinical laboratory technologist, the nephrologist identified more RBC (median of 80.000 vs 4.800, p= 0.001), more RBC casts (39.3% vs 0%, p=0.001), more urinary acanthocytes or G1 cells >5% (35.7% vs 7.14%, p=0.021) and more dysmorphic RBC (96.2% vs 7.14%,p<0.001). The discrepancies of the reports were maintained after the separation of the glomerulonephritis in proliferative and non proliferative. Conclusion: The urinary parameters used in characterization of the origin of the hematuria were more frequently identified by the nephrologist, and suggest that the urinalysis, a simple and very informative test, should be mandatory in programs of training in nephrology.

Clinical Significance of Urinary G1 Cells in the Differentiation of Hematuria / 대한신장학회잡지

BACKGROUND: Recently, G1 cells, characterized by distinctive doughnut-like shape with blebs have been reported as a reliable marker for glomerular hematuria. We investigated the validity of the urinary G1 cells in distingushing glomerular from non-glomerular hematuria. In addition, we evaluate the influence of urine osmolality, pH and proteinuria on dysmorphic erythrocytes and G1 cells. METHODS: One hundred and twenty patients with hematuria including 60 glomerular (GH) and 60 non- glomerular hematuria (NGH) were examined. The percentage of urinary dysmorphic erythrocytes and G1 cells using phase-contrast microscopy was determined. Urine osmolality, pH, and spot urine protein/ creatinine ratio were examined. RESULTS: The proportion of G1 cells differed significantly between the two group (7.8+/-16.0% in GH vs. 0% in NGH, p<0.05). At the cut-off value of 50 % dysmorphic erythrocytes, the sensitivity and specificity for the detection of GH was 88.3% and 93.3%, respectively. At the cut-off value of 1% G1 cells, sensitivity and specificity were 60.0% and 100%, respectively. When both of 50% dysmorphic erythrocytes and 1% G1 cells were considered as the cut-off value, the sensitivity and specificity were 91.0% and 100%, respectively. There was a significant difference in the percentage of dysmorphic erythrocytes and G1 cells at different urine pH. There was a significant correlation between urine osmolality and dysmorphic erythrocytes (r=0.41, p< 0.05), but not for G1 cells. No significant correlations were observed between G1 cells and proteinuria or pH. CONCLUSION: Evaluation of both urinary G1 cell and dysmorphic erythrocytes at the same time could improve the diagnostic value for differentiating glomerular hematuria.