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Objective:To study the effects of the conditioned medium (CM) from human lung adenocarcinoma cell line A549 on the viability and apoptosis of human umbilical vein endothelial cells (HUVECs) and the role of PI3K-Akt signaling pathway in the process. Methods:HUVECs were cultured with CM of A549 cells. Cell viability was detected by XTT assay. The morphological changes of HUVECs were analyzed by Hoechst 33258 staining and fluorescence microscopy. The apoptosis was detected by flow cytometry. Expression levels of total Akt and phosphorylated Akt were assessed by Western blotting. PI3K inhibitors wortmannin(WT)and Akt1 siRNA(siAkt1)were used to block PI3K-Akt signaling pathway. The mRNA transcription of Akt subtype was determined by RT-PCR.Results:A549 CM significantly increased cell viability after 24 h treatment (P=0.037) and inhibited apoptosis (P=0.001) of HUVECs. CM time-dependently activated phosphorylation of Akt. Akt was phosphorylated at 15 min after CM treatment and reached the peak at 30 min and then tended to decline. Both WT and siAkt1 blocked the effects of CM. Conclusion:The CM of A549 cells increased the survival and inhibit the apoptosis of HUVECs. Akt1 played a significant role in the process.
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Sixty-eight patients with pneumoconiosis combined with chronic obstructive pulmonary disease underwent large volume lavage in one lung under double cavity tracheal intubation and intravenousinhalant anesthesia. The vital signs of patients were recorded before, 10, 30min after and at the end of lavage. Results showed that the vital signs were stable during the lavage; and after the lavage all patients had relief significantly from the symptoms of dyspnea, polypnea and cough. Our results indicate that general anesthesia with bilateral lung ventilation are a safe and effective method in large volume whole lung lavage for treatment of pneumoconiosis patients combined with chronic obstructive pulmonary disease.
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AIM:To investigate the effects of high-fat diet on the level of intracellular free calcium ([Ca2+]i) and the activity of angiotensinⅠconverting enzyme (ACE) in alveolar macrophages (AMs) of rabbits. The association between asthma and high-fat diet was also observed. METHODS: Twelve male New Zealand rabbits were medially divided into normal diet group and 1.2% high-cholesterol diet group randomly. 8 weeks later, bronchial alveolar lavage was performed in vitro. [Ca2+]i was determined by Fluo-2/am.The activity of ACE was detected with ultraviolet method. RESULTS: The levels of [Ca2+]i in AMs greatly increased (P
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AIM: To evaluate the role of human angiotensin Ⅱ(AngⅡ) type Ⅰ receptor (AT1R) antisense cDNA (ahAT1) on migration of cultured artery smooth muscle cells (VSMCs). METHODS: Two recombinant adenoviral vectors, Ad/CMV. ahAT1 containing full length antisense cDNA targeting to human AT1R mRNA, and Ad/CMV.LacZ containing LacZ called report gene, were constructed by orientation clone technology and homologous recombination, and then were used to transfect VSMCs in vitro. AT1R expression detected by RT-PCR and immunohistochemistry, and migration of VSMCs measured by Boyden's Chamer methods, were compared between transfected and non- transfected VSMCs. RESULTS: Forty-eight hours after Ad/CMV. ahAT1 transfection, the level of AT1R mRNA decreased markedly (50% of control group), and AT1R protein expression was significantly less (P