Phosphorylated Hsp27 activates ATM-dependent p53 signaling and mediates the resistance of MCF-7 cells to doxorubicin-induced apoptosis

DNA damage activates p53 and its downstream target genes, which further leads to apoptosis or survival either by the cell cycle arrest or by DNA repair. In many tumors, the heat shock protein 27 (Hsp27) is expressed at high levels to provide protection against anticancer drugs. However, the roles of Hsp27 in p53-mediated cellular responses to DNA damage are controversial. Here, we investigated the interplay between the phosphorylation status of Hsp27 and p53 in kidney 293A (HEK293A) cells and found that over-expressing phosphorylated Hsp27 mimics (Hsp27-3D) activated p53/p21 in an ATM-dependent manner. In addition, incubation with doxorubicin (Dox), an anticancer drug, induced Hsp27 phosphorylation in human adenocarcinoma cells (MCF-7). In contrast, inhibition of Hsp27 phosphorylation retarded both p53 induction and p21 accumulation, and led to cell apoptosis. Furthermore, phosphorylated Hsp27 increased p53 nuclear importing and its downstream target gene expression such as p21 and MDM2, while de-phosphorylated Hsp27 impeded this procession. Taken together, our data suggest that Hsp27, in its phosphorylated or de-phosphorylated status, plays different roles in regulating p53 pathway and cell survival.     

谷子β-胡萝卜素异构酶家族基因的表达与变异分析

株型是影响谷类作物产量的重要性状, 株型改良对提高作物产量具有重要意义。独脚金内酯(strigolactones, SLs)作为一种最新被鉴定的植物激素, 其通过抑制腋芽的伸长调控分枝/分蘖的形成。β-胡萝卜素异构酶(D27s)是SLs合成途径的关键酶, 通过对谷子(Setaria italica) β-胡萝卜素异构酶典型结构域Pfam:DUF4033进行分析, 鉴定到3个谷子D27s基因家族成员(Seita.8G168400、Seita.6G088800和Seita.3G050900)。蛋白质特性分析显示, 谷子D27s蛋白由271-277个氨基酸残基组成, 分子量为30.1-30.4 kDa, 等电点为5.85-9.31, 不稳定系数介于38.48-74.47之间, 且均定位于叶绿体; 系统进化分析发现, 谷子D27s家族成员位于3个不同进化分支; 顺式作用元件预测显示, SiD27-1 (Seita.8G168400)可能参与调控生物节律、生长素介导的生长发育以及干旱和低温等胁迫应答过程。基因表达分析显示, SiD27-1在谷子多分蘖材料中表达下调, 在低磷胁迫处理下, D27s基因均能产生不同程度的响应, 并且SiD27-1的响应较其它成员更快速。单倍型分析结果表明, SiD27-1的H001单倍型为优异单倍型, 对谷子的株高、抽穗期和产量改良具有重要应用价值。综上, 推测SiD27-1极可能在SLs合成中发挥关键作用并对谷子株型产生影响。研究结果为深入揭示D27s对谷子分蘖形成的调控机制奠定了基础, 也为谷子株型分子设计育种提供了优异的等位变异位点。     

Celecoxib increases retinoid sensitivity in human colon cancer cell lines

Retinoid resistance has limited the clinical application of retinoids as differentiation-inducing and apoptosis-inducing drugs. This study was designed to investigate whether celecoxib, a selective COX-2 inhibitor, has effects on retinoid sensitivity in human colon cancer cell lines, and to determine the possible mechanism of said effects. Cell viability was measured using the MTT assay. Apoptosis was detected via Annexin-V/PI staining and the flow cytometry assay. PGE₂ production was measured with the ELISA assay. The expression of RARβ was assayed via western blotting. The results showed that celecoxib enhanced the inhibitory effect of ATRA in both COX-2 high-expressing HT-29 and COX-2 low-expressing SW480 cell lines. Further study showed the ATRA and celecoxib combination induced greater apoptosis, but that the addition of PGE₂ did not affect the enhanced growth-inhibitory and apoptosis-inducing effects of the combination. Moreover, NS398 (another selective COX-2 inhibitor) did not affect the inhibitory effects of ATRA in the two cell lines. Western blotting showed that the expression of RARβ in HT-29 cell lines was increased by celecoxib, but not by NS398, and that the addition of PGE₂ did not affect the celecoxib-induced expression of the retinoic acid receptor beta. In conclusion, celecoxib increased the expression of RARβ and the level of cellular ATRA sensitivity through COX-2-independent mechanisms. This finding may provide a potential strategy for combination therapy.     

Belowground processes and sustainability in agroecosystems with intercropping

Plant and Soil - Crop diversity has been repeatedly shown to support multiple ecosystem functions, both directly and indirectly, driven by interspecific root-root interactions. Despite continuous...     

On symmetric semiparametric two-sample problem

We consider a two-sample problem where data come from symmetric distributions. Usual two-sample data with only magnitudes recorded, arising from case-control studies or logistic discriminant analyses, may constitute a symmetric two-sample problem. We propose a semiparametric model such that, in addition to symmetry, the log ratio of two unknown density functions is modeled in a known parametric form. The new semiparametric model, tailor-made for symmetric two-sample data, can also be viewed as a biased sampling model subject to symmetric constraint. A maximum empirical likelihood estimation approach is adopted to estimate the unknown model parameters, and the corresponding profile empirical likelihood ratio test is utilized to perform hypothesis testing regarding the two population distributions. Symmetry, however, comes with irregularity. It is shown that, under the null hypothesis of equal symmetric distributions, the maximum empirical likelihood estimator has degenerate Fisher information, and the test statistic has a mixture of χ²-type asymptotic distribution. Extensive simulation studies have been conducted to demonstrate promising statistical powers under correct and misspecified models. We apply the proposed methods to two real examples.     

Effects of nitrogen addition and mowing on nitrogen- and water-use efficiency of Artemisia frigida in a grassland restored from an abandoned cropland

氮添加和刈割对内蒙古弃耕草地冷蒿氮和水分利用效率的影响 在氮和水分限制的区域，植物氮利用效率(NUE)和水分利用效率(WUE)决定了它们在群落中的竞争优势。冷蒿(Artemisia frigida)是半干旱草地重度退化的先锋物种，在不同退化程度的草地中具有不 同的优势度，经常被认为是退化草地群落演替的指示物种。退化草地恢复过程中，氮添加和割草如何影响冷蒿的NUE和WUE尚不清晰。以内蒙古多伦县弃耕草地为研究对象，选取两个不同群落斑块(禾草和冷蒿为优势物种的斑块)，经过长期(2006–2013)氮添加和刈割(对照、氮添加、刈割、氮添加+刈割)处理后，研究冷蒿的NUE (叶片碳氮比)和WUE (叶片碳同位素，δ¹³C)对氮添加、刈割及其交互作用的响应； 结合植物和土壤的碳、氮同位素(δ¹³C和δ¹⁵N)及碳、氮库探究退化草地恢复过程中植物的资源利用策略及其机制。研究结果表明：(1)氮添加对冷蒿的WUE没有显著影响(P > 0.05)，但NUE 在禾草和冷蒿斑块 中分别显著降低了42.9%和26.6% (P < 0.05)；(2)植物对不同氮源(NH₄₊或NO_3-)的利用会引起植物和土壤δ¹⁵N的分馏，研究表明叶片和土壤的δ¹⁵N与NUE呈现相反的变化趋势，因此冷蒿的NUE对氮添加的响应与不同氮源的利用有关；(3)刈割不影响冷蒿的NUE (P > 0.05)，但在禾草斑块，冷蒿的WUE在刈割处理下显著提高了2.3% (P < 0.05)；(4)在禾草斑块，氮添加减缓了割草对冷蒿WUE的促进作用；(5)结构方程模型显示，土壤含水量直接或间接的调控着冷蒿的WUE和NUE。综上所述，在禾草斑块，氮添加+刈割处理维持较低的NUE和WUE，不利于冷蒿对资源的竞争，进一步降低其优势度，这也预示着氮添加+ 刈割处理会促进退化草地的恢复。     

Kinetics of the course of reactivation of aminoacylase reconstituted using Mn2+ ions

The kinetic theory of the substrate reaction during irreversible change of enzyme activity previously described by Tsou (Tsou (1988),Adv. Enzymol. Relat. Areas Mol. Biol.61, 381–436] has been applied to a study of the kinetics of the course of reactivation during reconstitution of apo-aminoacylase using Mn²⁺ or Zn²⁺. The kinetic parameters for Mn²⁺-and Zn²⁺-reconstituted enzymes and the microscopic rate constants for reactivation during reconstitution were determined. The kinetic analysis suggests the presence of a second Mn²⁺ binding site in Mn²⁺-reconstituted aminoacylase.     

Subnetwork identification and chemical modulation for neural regeneration: A study combining network guided forest and heat diffusion model

Background: The induction of neural regeneration is vital to the repair of spinal cord injury (SCI). While compared with peripheral nervous system (PNS), the regenerative capacity of the central nervous system (CNS) is extremely limited. This indicates that modulating the molecular pathways underlying PNS repair may lead to the discovery of potential treatment for CNS injury.Methods: Based on the gene expression profiles of dorsal root ganglion (DRG) after a sciatic nerve injury, we utilized network guided forest (NGF) to rank genes in terms of their capacity of distinguishing injured DRG from sham-operated controls. Gene importance scores deriving from NGF were used as initial heat in a heat diffusion model (HotNet2) to infer the subnetworks underlying neural regeneration in the DRG. After potential regulators of the subnetworks were found through Connectivity Map (cMap), candidate compounds were experimentally evaluated for their capacity to regenerate the damaged neurons.Results: Gene ontology analysis of the subnetworks revealed ubiquinone biosynthetic process is crucial for neural regeneration. Moreover, almost half of the genes in these subnetworks are found to be related to neural regeneration via text mining. After screening compounds that are likely to modulate gene expressions of the subnetworks, three compounds were selected for the experiment. Of them, trichostatin A, a histone deacetylase inhibitor, was validated to enhance neurite outgrowth in vivo via an optic nerve crush mouse model.Conclusions: Our study identified subnetworks underlying neural regeneration, and validated a compound can promote neurite outgrowth by modulating these subnetworks. This work also suggests an alternative approach for drug repositioning that can be easily extended to other disease phenotypes.