18 research outputs found

    Comparison among BrdU labeling, Flow Cytometry and SiLAD to check cell cycle stage.

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    <p>Black line a means only to determine the cell cycle phase roughly. Curve line b means detecting the specific time point exactly. BrdU L is stands for the BrdU labeling and FCM is Flow Cytometry.</p

    Evaluation the synchronized efficiency of serum starved A549 cells.

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    <p>(A) FACS analysis of A549 cells synchronization by serum starvation for 48 hr. (B) Western blot analysis of expression of CyclinD1 at the indicate time lengths after serum re-stimulation. (C) FACS to detect the cell cycle progress after serum re-stimulation.</p

    SiLAD profiles are served to characterize the specific state of the cell cycle progress.

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    <p>Curve 1 was from the CBB spot in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002991#pone-0002991-g003" target="_blank">Figure 3D</a>. Curve 2 was from the spot in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002991#pone-0002991-g003" target="_blank">Figure 3G</a> PH-I image labeled with downward arrow. Curve 3 was from the same image with Curve 2, but was the spot labeled with upward arrow. The right panels A and B were the defined bar codes for the two time points labeled with spotted line A and B in the left panel. The curve was made by the software Origin6.</p

    Dynamic metabolized changes of the proteins.

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    <p>(A, D, G and I) Zoom in view of the spots dynamic changes at different time points. Panel A, D, and G show three differential changed proteins in A549 model. The differential protein shown in panel I was from rat liver hepatectomy model. PH-I stands for the images were got from Phosphor-imaging, while CBB means the CBB staining. (B and E) Shown the dynamic expression changes of the spots labeled in panel A and D, respectively. The x axis is time (t), and the y axis is the protein synthesizing speed (v<sub>expressed</sub>), and the area of each rectangle means the total amount of protein synthesized during each 15 minutes interval (s<sub>expressed</sub>). The midpoint of each rectangle's top edge was used to interpolate these five points with piecewise cubic Hermite polynomial, as shown in the diagram as the green line. (C and F) Shown the dynamic total amount changes of the spots labeled in A and D. The x axis also is time (t), and the y axis is the total protein metabolized speed (v<sub>existed</sub>), consequently the area of each rectangle means the total amount of protein (s<sub>existed</sub>) variation during each time interval. This green line indicates the function of the variation speed of the total amount of existed protein (s<sub>existed</sub>) with respect to time (t), during these six hours. (H) Presence %Volumes changes of the protein labeled in chart G. Ph-R denotes phosphorylation rate, <i>i.e.</i> the percentage of phosphorylated proteins accounted for the total sum in each time point. The value of PH-R represents mean±s.e.m. for 3 independent experiments. Error bars represent mean±s.e.m., n = 3. Chart B, C, E and F were made by the software MatLab, and chart H was made by SPSS13.</p

    The Characteristics and Function of S100A7 Induction in Squamous Cell Carcinoma: Heterogeneity, Promotion of Cell Proliferation and Suppression of Differentiation

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    <div><p>S100A7 is highly expressed in squamous cell carcinomas (SCC) and is related to the terminal differentiation of keratinocytes. However, its characteristic and function in SCC is not very known. In this present study, we used immunohistochemistry to examine the expression of S100A7 in 452 SCC specimens, including the lung, esophagus, oral cavity, skin, cervix, bladder, and three SCC cell lines. We found that S100A7-positive staining showed significant heterogeneity in six types of SCC specimen and three SCC cell lines. Further examination found that S100A7-positive cells and its expression at mRNA and protein levels could be induced in HCC94, FaDu, and A-431 cells both in vitro and in vivo using immunohistochemistry, real-time PCR, and Western blotting. Notably, the upregulation of squamous differentiation markers, including keratin-4, keratin-13, TG-1, and involucrin, also accompanied S100A7 induction, and a similar staining pattern of S100A7 and keratin-13 was found in HCC94 cells both in vitro and in vivo. Further study revealed that the overexpression of S100A7 significantly increased proliferation and inhibited squamous differentiation in A-431 cells both in vitro and in vivo. Conversely, silencing S100A7 inhibited cell growth and survival and increased the expression of keratin-4, keratin-13, TG-1, and involucrin in HCC94 cells. Therefore, these results demonstrate that S100A7 displays heterogeneous and inducible characteristic in SCC and also provide novel evidence that S100A7 acts as a dual regulator in promoting proliferation and suppressing squamous differentiation of SCC.</p></div

    The precautionary principle and genetically modified organisms: a bone of contention between European institutions and member states

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    Theoretical thesis.Bibliography: pages 75-86.Introduction -- Chapter I. The precautionary principle -- Chapter II. The precautionary principle and genetically modified organisms under the European legal framework -- Chapter III. The precautionary principle and genetically modified organisms : analysis of three seminal cases of the European Court of Justice (2003-2011) -- Chapter IV. A high level of protection for health and the environment vs free circluation of GMOs on the European market : interests in conflict -- Bibliography.This dissertation examines how the Precautionary Principle, as an internationally recognised concept enshrined in a range of legal instruments, has been applied to provide a mechanism for protection of the environment and health in response to the introduction of Genetically Modified Organisms (GMOs) in Europe. It examines how the European Court of Justice substantively handled the risk assessment phase across three seminal cases between 2003 and 2011 in which Member States had failed in their attempt to trigger the Precautionary Principle in order to uphold a ban or suspension of the cultivation or sale of products derived from GMOs in their territory. The analysis of these judgements suggests that the Court has applied a narrow approach to the evidence provided by national governments during the risk assessment stage, and has thereby limited the potential for precautionary measures by Member States to be upheld by the Court. This outcome reflects a ‘weak’ application of the Precautionary Principle by the Court in contrast with the ‘strong’ interpretation implied by the European legal and policy framework and objections of Member States.Mode of access: World wide web1 online resource (86 pages

    The expression of S100A7 in SCC tissues.

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    <p>SCC and normal tissues were examined by immunohistochemistry with specific anti-S100A7 antibody in lung (A, M); esophagus (B, N); cervix (C, O); bladder (D, P); oral cavity (E, Q); skin (F, R). The corresponding types of SCC tissues were also examined by immunohistochemistry with nonspecific IgG (G-L). Arrowheads indicate the positive staining of S100A7 and the asterisks indicate the keratinizing areas.</p

    TEAD1 mediates YAP-dependent S100A7 expression.

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    <p>(a) Western blot analyses for S100A7 expression after TEAD1 silencing in normal attached HCC94 cells. (b) The mRNA expression of <i>TEAD1</i>, <i>CTGF</i>, <i>CYR61</i> and <i>S100A7</i> is examined using qRT-PCR. H: HCC94 cells. Error bar, SD of three different experiments. <i>*P<0</i>.<i>05</i>, **<i>P</i><0.01; <i>t-test</i>. (c, d) Overexpression of YAP-WT and mutant activated YAP-S94A in normal attached HCC94 (c) and FaDu (d) cells. Anti-Flag tag antibody was used to judge the transfection efficiency. β-actin was used as a loading control.</p
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