117 research outputs found

    Hypoxia-inducible transcription factor-1α promotes hypoxia-induced A549 apoptosis via a mechanism that involves the glycolysis pathway

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    BACKGROUND: Hypoxia-inducible transcription factor-1α (HIF-1α), which plays an important role in controlling the hypoxia-induced glycolysis pathway, is a "master" gene in the tissue hypoxia response during tumor development. However, its role in the apoptosis of non-small cell lung cancer remains unknown. Here, we have studied the effects of HIF-1α on apoptosis by modulating HIF-1α gene expression in A549 cells through both siRNA knock-down and over-expression. METHODS: A549 cells were transfected with a HIF-1α siRNA plasmid or a HIF-1α expression vector. Transfected cells were exposed to a normoxic or hypoxic environment in the presence or absence of 25 mM HEPES and 2-deoxyglucose (2-DG) (5 mM). The expression of three key genes of the glycolysis pathway, glucose transporter type 1(GLUT1), phosphoglycerate kinase 1(PGK1), and hexokinase 1(HK1), were measured using real-time RT-PCR. Glycolysis was monitored by measuring changes of pH and lactate concentration in the culture medium. Apoptosis was detected by TUNEL assay and flow cytometry. RESULTS: Knocking down expression of HIF-1α inhibited the glycolysis pathway, increased the pH of the culture medium, and protected the cells from hypoxia-induced apoptosis. In contrast, over-expression of HIF-1α accelerated glycolysis in A549 cells, decreased the pH of the culture medium, and enhanced hypoxia-induced apoptosis. These effects of HIF-1α on glycolysis, pH of the medium, and apoptosis were reversed by treatment with the glycolytic inhibitor, 2-DG. Apoptosis induced by HIF-1α over-expression was partially inhibited by increasing the buffering capacity of the culture medium by adding HEPES. CONCLUSION: During hypoxia in A549 cells, HIF-1α promotes activity of the glycolysis pathway and decreases the pH of the culture medium, resulting in increased cellular apoptosis

    Higher-order assembly of crystalline cylindrical micelles into membrane-extendable colloidosomes

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    Functional nanoscale objects can be prepared via crystallization-driven self-assembly of diblock copolymers. Here the authors show the self-assembly of crystalline block copolymers into size-specific cylindrical micelles for the hierarchical construction of mechanically robust colloidosomes with a range of membrane textures

    Acute extensive portal vein thrombosis with pulmonary embolism: a case report

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    Which user errors matter during HIV self-testing? A qualitative participant observation study of men who have sex with men (MSM) in China

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    Abstract Background The World Health Organization recommends HIV self-testing (HIVST) as an additional approach to HIV testing services. We aimed to assess to what extent HIVST was conducted correctly by Chinese men who have sex with men (MSM) and to identify user errors during the HIVST process in order to inform strategies to optimize its use and thus reduce the number of undiagnosed HIV infections. Methods Between February and March 2017, participant observations were conducted with 27 MSM in an east coastal city in China. In the presence, but without the assistance or orientation, of a trained HIV testing counselor, participants conducted HIVST (either finger prick or oral fluid) according to manufacturers’ instructions. Errors were recorded on checklists during direct observation and double checked afterwards by reviewing video files of the observations. Results Overall, 12 participants (44.4%) had invalid test results due to user errors. Just five (18.5%) did not make any errors during the entire HIVST process. Failure to follow all the steps based on manufactures’ instructions was a common problem for both finger prick and oral fluid self-testers. For finger prick users, most errors occurred during the stage of collecting the specimen. In contrast, oral fluid users made most errors during the stage of testing the collected specimen. Conclusions Although we found that user errors were common among MSM administering HIVST, this should not deter or discourage routine implementation and scale-up of HIVST as strategies can be implemented to facilitate the correct use of HIVST. Trial registration This study was a part of a clinical trial: ClinicalTrials.gov (#NCT02999243); Registration date: December 20, 2016
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