OBJECTIVE: To evaluate any difference in levels of umbilical venous eryhthropoietin (EPO) and nucleated red blood cells (NRBC) between appropriate for gestational age (AGA) and small for gestational age (SGA) preterm neonates at birth and to evaluate the peripartal factors that influence the secretion of the nucleated red blood cells in preterm neonate. METHODS: 43 preterm singleton neonates born at the gestational age between 27 weeks and 37 weeks of gestation from January 1998 to December 2004 were enrolled and divided into 25 cases of AGA and 18 cases of SGA. At each delivery, umbilical venous blood gas values, concentration of EPO by radioimmunoassay and the NRBC count expressed per 100 white blood cell (WBC) were obtained. The placenta were examined microscopically for the presence of pathologic infarct and inflammation. Statistical analysis was done by Mann-Whitney U test, Fisher exact test, univariate and multiple regression analysis using SPSS statistical package. RESULTS: The median umbilical venous EPO concentration and fetal hemoglobin level in SGA preterm neonates were 48.0 mIU/mL and 15.7 g/dL, which were significantly higher than those in AGA preterm neonates (12.5 mIU/ML, 14.6 g/dL). The median NRBC in SGA group was 8.0 NRBC/100 WBC which was higher than in the AGA group (2.5 NRBC/100 WBC), showing no significant difference between groups. Stepwise multiple regression analysis identified O2 saturation, emergency cesarian section, infarct and inflammation in placental pathology and premature rupture of membranes as independent variables associated with the NRBC count. CONCLUSION: Measurement of the level of EPO and NRBC in umbilical venous blood at birth of the preterm neonates can be used as a helpful index for evaluation of intrauterine hypoxia. In addition, cord blood gas ananlysis and placental examination on the infarct and inflammation are informative value for the elevated NRBC.
Anoxia ; Emergencies ; Erythroblasts ; Erythrocytes ; Erythropoietin ; Fetal Blood ; Fetal Hemoglobin ; Gestational Age ; Humans ; Infant, Newborn ; Inflammation ; Leukocytes ; Membranes ; Parturition ; Placenta ; Pregnancy ; Radioimmunoassay ; Rupture

OBJECTIVE: To evaluate any difference in levels of umbilical venous eryhthropoietin (EPO) and nucleated red blood cells (NRBC) between appropriate for gestational age (AGA) and small for gestational age (SGA) preterm neonates at birth and to evaluate the peripartal factors that influence the secretion of the nucleated red blood cells in preterm neonate. METHODS: 43 preterm singleton neonates born at the gestational age between 27 weeks and 37 weeks of gestation from January 1998 to December 2004 were enrolled and divided into 25 cases of AGA and 18 cases of SGA. At each delivery, umbilical venous blood gas values, concentration of EPO by radioimmunoassay and the NRBC count expressed per 100 white blood cell (WBC) were obtained. The placenta were examined microscopically for the presence of pathologic infarct and inflammation. Statistical analysis was done by Mann-Whitney U test, Fisher exact test, univariate and multiple regression analysis using SPSS statistical package. RESULTS: The median umbilical venous EPO concentration and fetal hemoglobin level in SGA preterm neonates were 48.0 mIU/mL and 15.7 g/dL, which were significantly higher than those in AGA preterm neonates (12.5 mIU/ML, 14.6 g/dL). The median NRBC in SGA group was 8.0 NRBC/100 WBC which was higher than in the AGA group (2.5 NRBC/100 WBC), showing no significant difference between groups. Stepwise multiple regression analysis identified O2 saturation, emergency cesarian section, infarct and inflammation in placental pathology and premature rupture of membranes as independent variables associated with the NRBC count. CONCLUSION: Measurement of the level of EPO and NRBC in umbilical venous blood at birth of the preterm neonates can be used as a helpful index for evaluation of intrauterine hypoxia. In addition, cord blood gas ananlysis and placental examination on the infarct and inflammation are informative value for the elevated NRBC.
Anoxia ; Emergencies ; Erythroblasts ; Erythrocytes ; Erythropoietin ; Fetal Blood ; Fetal Hemoglobin ; Gestational Age ; Humans ; Infant, Newborn ; Inflammation ; Leukocytes ; Membranes ; Parturition ; Placenta ; Pregnancy ; Radioimmunoassay ; Rupture

BACKGROUND: Autophagy is an important adaptive mechanism in normal development and in response to changing environmental stimuli in cancer. Previous papers have reported that different types of cancer underwent autophagy to obtain amino acids as energy source of dying cells in nutrient-deprived conditions. However, whether or not autophagy in the process of lung cancer causes death or survival is controversial. Therefore in this study, we investigated whether nutrient deprivation induces autophagy in human H460 lung cancer cells. METHODS: H460, lung cancer cells were incubated in RPMI 1640 medium, and the starved media, which are BME and RPMI media without serum, including 2-deoxyl-D-glucose according to time dependence. To evaluate the viability and find out the mechanism of cell death under nutrient-deprived conditions, the MTT assay and flow cytometry were done and analyzed the apoptotic and autophagic related proteins. It is also measured the development of acidic vascular organelles by acridine orange. RESULTS: The nutrient-deprived cancer cell is relatively sensitive to cell death rather than normal nutrition. Massive cytoplasmic vacuolization was seen under nutrient-deprived conditions. Autophagic vacuoles were visible at approximately 12 h and as time ran out, vacuoles became larger and denser with the increasing number of vacuoles. In addition, the proportion of acridine orange stain-positive cells increased according to time dependence. Localization of GFP-LC3 in cytoplasm and expression of LC-3II and Beclin 1 were increased according to time dependence on nutrient-deprived cells. CONCLUSION: Nutrient deprivation induces cell death through autophagy in H460 lung cancer cells.
Acridine Orange ; Amino Acids ; Autophagy ; Cell Death ; Cytoplasm ; Flow Cytometry ; Humans ; Lung Neoplasms ; Malnutrition ; Organelles ; Proteins ; Vacuoles

BACKGROUND: Most lung cancer patients receive systemic chemotherapy at an advanced stage disease. Cisplatin-based chemotherapy is the main regimen for treating advanced lung cancer. Recently, autophagy has become an important mechanism of cellular adaptation under starvation or cell oxidative stress. The purpose of this study was to determine whether or not autophagy can occurred in cisplatin-treated lung cancer cells. METHODS: H460 cells were incubated with RPMI 1640 and treated in 5 micrometer or 20 micrometer cisplatin concentrations at specific time intervals. Cells surviving cisplatin treatment were measured and compared using an MTT cell viability assay to cells that underwent apoptosis with autophagy by nuclear staining, apoptotic or autophagic related proteins, and autophagic vacuoles. The development of acidic vascular organelles was using acridine orange staining and fluorescent expression of GFP-LC3 protein in its transfected cells was observed to evaluate autophagy. RESULTS: Lung cancer cells treated with 5 micrometer cisplatin-treated were less sensitive to cell death than 20 micrometer cisplatin-treated cells in a time-dependent manner. Nuclear fragmentation at 5 micrometer was not detected, even though it was discovered at 20 micrometer. Poly (ADP-ribose) polymerase cleavages were not detected in 5 micrometer within 24 hours. Massive vacuolization in the cytoplasm of 5 micrometer treated cells were observed. Acridine orange stain-positive cells was increased according in time-dependence manner. The autophagosome-incorporated LC3 II protein expression was increased in 5 micrometer treated cells, but was not detected in 20 micrometer treated cells. The expression of GFP-LC3 were increased in 5 micrometer treated cells in a time-dependent manner. CONCLUSION: The induction of autophagy occurred in 5 micrometer dose of cisplatin-treated lung cancer cells.
Acridine Orange ; Apoptosis ; Autophagy ; Cell Death ; Cell Survival ; Cisplatin ; Cytoplasm ; Humans ; Lung ; Lung Neoplasms ; Organelles ; Oxidative Stress ; Proteins ; Starvation ; Vacuoles

BACKGROUND: Most lung cancer patients receive systemic chemotherapy at an advanced stage disease. Cisplatin-based chemotherapy is the main regimen for treating advanced lung cancer. Recently, autophagy has become an important mechanism of cellular adaptation under starvation or cell oxidative stress. The purpose of this study was to determine whether or not autophagy can occurred in cisplatin-treated lung cancer cells. METHODS: H460 cells were incubated with RPMI 1640 and treated in 5 micrometer or 20 micrometer cisplatin concentrations at specific time intervals. Cells surviving cisplatin treatment were measured and compared using an MTT cell viability assay to cells that underwent apoptosis with autophagy by nuclear staining, apoptotic or autophagic related proteins, and autophagic vacuoles. The development of acidic vascular organelles was using acridine orange staining and fluorescent expression of GFP-LC3 protein in its transfected cells was observed to evaluate autophagy. RESULTS: Lung cancer cells treated with 5 micrometer cisplatin-treated were less sensitive to cell death than 20 micrometer cisplatin-treated cells in a time-dependent manner. Nuclear fragmentation at 5 micrometer was not detected, even though it was discovered at 20 micrometer. Poly (ADP-ribose) polymerase cleavages were not detected in 5 micrometer within 24 hours. Massive vacuolization in the cytoplasm of 5 micrometer treated cells were observed. Acridine orange stain-positive cells was increased according in time-dependence manner. The autophagosome-incorporated LC3 II protein expression was increased in 5 micrometer treated cells, but was not detected in 20 micrometer treated cells. The expression of GFP-LC3 were increased in 5 micrometer treated cells in a time-dependent manner. CONCLUSION: The induction of autophagy occurred in 5 micrometer dose of cisplatin-treated lung cancer cells.
Acridine Orange ; Apoptosis ; Autophagy ; Cell Death ; Cell Survival ; Cisplatin ; Cytoplasm ; Humans ; Lung ; Lung Neoplasms ; Organelles ; Oxidative Stress ; Proteins ; Starvation ; Vacuoles