RESUMEN
BACKGROUND: Primary immune thrombocytopenic purpura (pITP) is defined as isolated autoimmune thrombocytopenia with idiopathic low platelet count, normal bone marrow, and unexplained causes of thrombocytopenia. Currently there is no definite criterion for ITP diagnosis. METHODS: We conducted proteomic screen of patients with pITP, secondary immune thrombocytopenia (sITP), and healthy controls using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). The proteomic profiles were obtained from platelet lysate samples of 82 healthy adult controls, 64 pITP, and 70 sITP patients, from which we screened marker proteins with significant differences, and constructed a diagnosis model using the artificial neural network (ANN) technique. RESULTS: We identified 6 marker proteins in the platelet lysates of pITP patients. This diagnosis method differentiated pITP patients from sITP effectively with a sensitivity of 96.9% (31/32), a specificity of 71.0% (54/76), and the area under the ROC curve of 0.864 in the training set, and a sensitivity of 87.5% (28/32), a specificity of 69.7% (53/76), and a positive predictive value of 75.0% (81/108) in the test set. CONCLUSION: The artificial neural network model based on platelet protein profiling established a potential pITP diagnosis platform.
Asunto(s)
Plaquetas/metabolismo , Proteómica , Púrpura Trombocitopénica Idiopática/sangre , Adulto , Biomarcadores/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Púrpura Trombocitopénica Idiopática/diagnóstico , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
AIM: To investigate protective effect and possible mechanisms of erythropoitin (EPO) on cortical neuron against damage induced by glutamate(Glu). METHODS: Cortical neurons were removed from 1-day-old newborn rat and then cultured in vitro. The model of damage induced by Glu was established. The experiment was designed as: control group, Glu treatment group,the group of pretreatment with EPO, pyrrolidine dithiocarbamates(PDTC) treatment group. The changes of morphology were observed under inverted microscopy; the cell viability was detected by MTT assay; and the apoptosis rate of neuron was measured by flow cytometry. RESULTS: After exposure to Glu, neurons were obviously damaged. Neurons morph of EPO pretreatment group was similar to that of blank group. However, in PDTC treatment group, cells were obviously damaged, and morph of cells was similar to that of Glu treatment group. Compared with control group, survival of neurons of Glu treatment group significantly decreased, while the neurons apoptotis of Glu treatment group was significantly increased (P<0h01). Compared with Glu treatment group, the neurons, survival of EPO pretreatment group significantly increased, the neurons apoptotis can significantly decrease (P<0h01)ls. Compared with EPO pretreatment group, the cells survival of PDTC treatment group was significantly decreased, while the neurons, apoptotis of Glu treatment group was significantly increased (P<0h01), and similar to that of Glu treatment group. CONCLUSION: EPO could protect cortical neuron from Glu toxicity,which might related to signal transduction of NF-kappaB.
Asunto(s)
Ácido Glutámico/toxicidad , FN-kappa B/metabolismo , Neuronas/efectos de los fármacos , Quinolinas/farmacología , Animales , Apoptosis/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Corteza Cerebral/citología , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/metabolismo , Neuronas/citología , Neuronas/metabolismo , Ratas , Ratas WistarRESUMEN
OBJECTIVE: To replicate a new model of injury to human renal proximal tubular cells (HK-2) induced by hypoxia/reoxygenation. METHODS: Human renal proximal tubular cell line HK-2 cell was used as the target cell. Tubular cells were divided into six groups: 4 hours of hypoxia, 12 hours of hypoxia, 24 hours of hypoxia, and 24 hours of hypoxia followed by reoxygenation 4, 12 or 24 hours later groups. Each group was accompanied by a control group. Hypoxic culture conditions were produced by covering the culture with liquid paraffin. Trypan blue exclusion was used for cell count and cell viability. The activity of lactate dehydrogenase (LDH) in the culture medium was determined by biochemical method. RESULTS: After being challenged by hypoxia followed by reoxygenation, trypan blue exclusion rate was greater, cell count and cell viability were lower, and the activity of LDH was increased. It indicated that the destruction of integrity of cellular membrane was induced by ischemia/reperfusion injury, and the tubular cells may be injured irreversibly. CONCLUSION: A simple model of hypoxic injury of renal tubular cells is replicated by covering the culture cells with liquid paraffin.
Asunto(s)
Hipoxia de la Célula , Túbulos Renales Proximales/citología , Línea Celular , Supervivencia Celular , Humanos , Túbulos Renales Proximales/enzimología , L-Lactato Deshidrogenasa/metabolismoRESUMEN
AIM: To investigate the relationship between TNF-alpha and renal tubular cell injury caused by anoxia/reoxygenation. METHODS: Human renal proximal tubular cell line HK-2 was used as model. Anoxia/reoxygenation were produced by covering/de-covering the cell culture with liquid paraffin wax. The level of TNF-alpha and the activity of lactate dehydrogenase(LDH) in the culture medium was determined by radioimmunoassay(RIA) and biochemical methods, respectively. Trypan blue exclusion was used to measure cell viability. RESULTS: Anoxia/reoxygenation could increase TNF-alpha level and LDH activity, but decrease viability of HK-2 cells. TNF-alpha level was positively correlated with LDH activity (r=0.89, P<0.05) and negatively with the cell viability (r=-0.91, P<0.05). CONCLUSION: TNF-alpha induced by anoxia/reoxygenation may participate in the process of renal tubular cell injury.