Protein kinase D inhibitor CRT0066101 suppresses bladder cancer growth in vitro and xenografts via blockade of the cell cycle at G2/M

The protein kinase D (PKD) family of proteins are important regulators of tumor growth, development, and progression. CRT0066101, an inhibitor of PKD, has antitumor activity in multiple types of carcinomas. However, the effect and mechanism of CRT0066101 in bladder cancer are not understood. In the present study, we show that CRT0066101 suppressed the proliferation and migration of four bladder cancer cell lines in vitro. We also demonstrate that CRT0066101 blocked tumor growth in a mouse flank xenograft model of bladder cancer. To further assess the role of PKD in bladder carcinoma, we examined the three PKD isoforms and found that PKD2 was highly expressed in eight bladder cancer cell lines and in urothelial carcinoma tissues from the TCGA database, and that short hairpin RNA (shRNA)-mediated knockdown of PKD2 dramatically reduced bladder cancer growth and invasion in vitro and in vivo, suggesting that the effect of the compound in bladder cancer is mediated through inhibition of PKD2. This notion was corroborated by demonstrating that the levels of phospho-PKD2 were markedly decreased in CRT0066101-treated bladder tumor explants. Furthermore, our cell cycle analysis by flow cytometry revealed that CRT0066101 treatment or PKD2 silencing arrested bladder cancer cells at the G2/M phase, the arrest being accompanied by decreases in the levels of cyclin B1, CDK1 and phospho-CDK1 (Thr161) and increases in the levels of p27^Kip1 and phospho-CDK1 (Thr14/Tyr15). Moreover, CRT0066101 downregulated the expression of Cdc25C, which dephosphorylates/activates CDK1, but enhanced the activity of the checkpoint kinase Chk1, which inhibits CDK1 by phosphorylating/inactivating Cdc25C. Finally, CRT0066101 was found to elevate the levels of Myt1, Wee1, phospho-Cdc25C (Ser216), Gadd45α, and 14-3-3 proteins, all of which reduce the CDK1-cyclin B1 complex activity. These novel findings suggest that CRT0066101 suppresses bladder cancer growth by inhibiting PKD2 through induction of G2/M cell cycle arrest, leading to the blockade of cell cycle progression.     

创新策略、设计策略与设计特色——以消费性电子产业为例

创新是企业成长的动力,而设计可以视为企业一项整合资源,也是产品开发成功的关键因素。但在企业实际产品研发过程中,设计要如何运作,才能达到企业创新的目标,是值得探讨的议题。在过去的研究中发现企业在产品设计方面,的确有一些具体的作法。但是实证企业在创新策略与设计策略乃至具体呈现设计特色之关联模型的研究却非常有限。本研究综合过去有关创新策略、设计策略以及设计特色之文献,针对新产品研发部门主管进行问卷调查。再以结构方程模式检验理论模式和观察数据的适配度。研究结果发现:创新策略、设计策略以及设计特色之关联理论模式和观察数据适配。企业之创新策略会影响产品设计特色,同时企业之创新策略会透过设计策略而影响产品设计特色,其中设计策略对设计特色而言,既是自变项又是中介变项。企业经营者所面临的挑战,往往是如何更快速地推出迎合消费市场需求之新产品,并且有效获利。本研究为首次针对这些创新设计策略变项所进行的实证研究,后续将再针对产品创新设计的议题进一步深入研讨。     

Chest wall abscess due to Prevotella bivia

Prevotella bivia is associated with pelvic inflammatory disease. A 77-year-old man developed a rapidly growing chest wall abscess due to P. bivia within days. He underwent surgical resection of the infected area; his postoperative course was uneventful. This is the first case of chest wall abscess due to P. bivia infection. Its correct diagnosis cannot be underestimated because fulminant infections can occur in aged or immunocompromised patients if treated incorrectly. Prompt, appropriate surgical management, and antibiotic therapy affect treatment outcome.     

A Meta‐learning Framework for Bankruptcy Prediction

The implication of corporate bankruptcy prediction is important to financial institutions when making lending decisions. In related studies, many bankruptcy prediction models have been developed based on some machine‐learning techniques. This paper presents a meta‐learning framework, which is composed of two‐level classifiers for bankruptcy prediction. The first‐level multiple classifiers perform the data reduction task by filtering out unrepresentative training data. Then, the outputs of the first‐level classifiers are utilized to create the second‐level single (meta) classifier. The experiments are based on five related datasets and the results show that the proposed meta‐learning framework provides higher prediction accuracy rates and lower type I/II errors when compared with the stacked generalization classifier and other three widely developed baselines, such as neural networks, decision trees, and logistic regression. Copyright © 2011 John Wiley & Sons, Ltd.     

《类经》中的藏象与以身体为中心的现代中医学的趋向

文章把人类文化学实地考察与教科书研究结合在一起。关于实地考察方面，作者在云南中医学院既是现场观察者，又是现场体验者。当时（１９８８～１９８９年）作者是第一个和唯一的外国学生，用中文学习中医学，与大学本科生一道上课和参加临床学习。关于教科书研究方面，作者利用了一些中医学教科书，主要研究这些教科书以及视为其前身的著作中叙述知识所依照的结构。教科书的研究表明，上述著作中叙述知识的结构在过去３００年里基本     

Adriamycin-induced chromosome damage: elevated frequencies of isochromatid aberrations in G2 and S phases.

Analysis of chromosomes from cells treated with adriamycin during G2 and S phases showed a high frequency of isochromatid-type of breaks, in addition to the expected chromatid breaks. These are interpreted as independent breaks on sister chromatids because of preferential effects of the drug in specific chromosome regions. The break points are likely to be different, but morphologically such breaks would be indistinguishable from isochromatid or chromosome breaks.     

A recycling pathway between the endoplasmic reticulum and the Golgi apparatus for retention of unassembled MHC class I molecules

Assembly of class I major histocompatibility complex (MHC) molecules involves the interaction of two distinct polypeptides (the heavy and light chains) with peptide antigen. Cell lines synthesizing both chains but expressing low levels of MHC class I molecules on their surface as a result of a failure in assembly and transport have been identified. We now report that although the apparent steady-state distribution in these cells of class I molecules is in the endoplasmic reticulum (ER), the molecules in fact are recycled between the ER and Golgi, rather than retained in the ER. This explains the failure of class I molecules to negotiate the secretory pathway. Class I molecules do not seem to be modified by Golgi enzymes, suggesting that the proteins do not reach the Golgi apparatus during recycling. But morphological and subcellular fractionation evidence indicates that they pass through the cis Golgi or a Golgi-associated organelle, which we postulate to be the recycling organelle. This compartment, which we call the 'cis-Golgi network', would thereby be a sorting organelle that selects proteins for return to the ER.     

Cell-surface antigens expressed on L-cells transfected with whole DNA from non-expressing and expressing cells

We have shown previously that transfection of mouse L-cells with DNA from JM, a human T-cell line expressing certain T-cell differentiation antigens, yields stable transfectants expressing one or another of these antigens. The identities of the antigens were confirmed by immunoprecipitation and SDS-polyacrylamide gel electrophoresis. We now report that our procedure--co-transfection with the chicken thymidine kinase gene (tk) and whole cellular DNA, selection with hypoxanthine-aminopterin-thymidine (HAT), and staining of the cells with fluorochrome-conjugated monoclonal antibodies and fluorescence-activated cell-sorter (FACS) selection--yields transfectants expressing a variety of cell-surface molecules (19 of 21 investigated), most at a frequency of about one per 10(3) Tk+ transformants. Of these, 9 of 12 were transferred and expressed as readily using DNA from cells which did not express the cell-surface antigens as from tissues or cells that did express them.     

Resolution of synthetic att-site Holliday structures by the integrase protein of bacteriophage lambda

Site-specific recombination of the bacteriophage lambda genome into and out of the host bacterial genome is postulated to involve the formation of Holliday structure intermediates by reciprocal single-strand exchanges. Synthetic analogues of the predicted recombination intermediates are resolved in vitro by the protein product of the lambda int gene. Some of the structural features and reaction conditions for this genetic recombination can now be defined.