An atypical Phytophthora sojae RxLR effector manipulates host vesicle trafficking to promote infection

In plants, the apoplast is a critical battlefield for plant-microbe interactions. Plants secrete defense-related proteins into the apoplast to ward off the invasion of pathogens. How microbial pathogens overcome plant apoplastic immunity remains largely unknown. In this study, we reported that an atypical RxLR effector PsAvh181 secreted by Phytophthora sojae, inhibits the secretion of plant defense-related apoplastic proteins. PsAvh181 localizes to plant plasma membrane and essential for P. sojae infection. By co-immunoprecipitation assay followed by liquid chromatography-tandem mass spectrometry analyses, we identified the soybean GmSNAP-1 as a candidate host target of PsAvh181. GmSNAP-1 encodes a soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein, which associates with GmNSF of the SNARE complex functioning in vesicle trafficking. PsAvh181 binds to GmSNAP-1 in vivo and in vitro. PsAvh181 interferes with the interaction between GmSNAP-1 and GmNSF, and blocks the secretion of apoplastic defense-related proteins, such as pathogenesis-related protein PR-1 and apoplastic proteases. Taken together, these data show that an atypical P. sojae RxLR effector suppresses host apoplastic immunity by manipulating the host SNARE complex to interfere with host vesicle trafficking pathway.     

模拟不同降雨时间格局下喀斯特垂直异质生境对桢楠幼苗光合和生长的影响

由于岩石裸露、地表破碎、坡度陡峻、洼地土壤堆积等因素的共同作用,喀斯特区域土壤具有明显的不连续性和高度异质性,土壤侵蚀往往较为严重。喀斯特地区高度异质性的生境为植物提供了不同的水分、肥料、空气和空间,对植物的生长和生理具有显著影响。在全球气候变化下,降雨时间格局的改变可能将进一步加剧喀斯特的生境异质性。桢楠是我国特有的国家二级保护树种,虽然它只零星分布在喀斯特地区,但由于它能在岩石间生长得很好,仍被认为是一种可能的适宜树种。以二年生桢楠（Phoebe zhennan S.Lee）幼苗为研究对象,研究总降水量相同条件下两种降雨时间格局（I_2d：2 d降雨间隔;I_19d：：19 d降雨间隔处理）和三种垂直异质小生境（S₀：无石全土,S_1/2：半石半土,S_3/4：多石少土）对其光合生理和生长的影响。结果显示：（1）在I_2d条件下,桢楠幼苗的光合速率在半石半土生境（S_1/2）下最大,且有较大的地径和株高;而在I_19d条件下,桢楠的光合和生长（株高和地径）均随着岩溶裂隙层的增厚逐渐减小。（2）在全土生境（S₀）下,降雨时间格局的改变并不影响桢楠的光合和株高,而在有岩溶裂隙层的生境下（S_1/2和S_3/4）,降雨时间间隔的延长会抑制桢楠的光合和株高及地径的生长,提高气孔密度,且这种抑制或促进作用会随着岩溶裂隙层的增厚而加剧。研究表明,在降雨间隔时间短,降雨均匀充足时,有少量的岩溶裂隙能够促进桢楠的生长和光合,但降雨间隔时间延长后,这种作用丧失。另一方面,在土壤相对较厚时,降雨时间间隔的改变对桢楠的光合和生长影响不显著,但岩溶裂隙层的增加,加剧了降雨时间间隔增大对桢楠光合的负面影响。     

Induction of Neuroendocrine Differentiation in Prostate Cancer Cells by Dovitinib (TKI-258) and its Therapeutic Implications

Prostate cancer (PCa) remains the second-leading cause of cancer-related deaths in American men with an estimated mortality of more than 26,000 in 2016 alone. Aggressive and metastatic tumors are treated with androgen deprivation therapies (ADT); however, the tumors acquire resistance and develop into lethal castration resistant prostate cancer (CRPC). With the advent of better therapeutics, the incidences of a more aggressive neuroendocrine prostate cancer (NEPC) variant continue to emerge. Although de novo occurrences of NEPC are rare, more than 25% of the therapy-resistant patients on highly potent new-generation anti-androgen therapies end up with NEPC. This, along with previous observations of an increase in the number of such NE cells in aggressive tumors, has been suggested as a mechanism of resistance development during prostate cancer progression. Dovitinib (TKI-258/CHIR-258) is a pan receptor tyrosine kinase (RTK) inhibitor that targets VEGFR, FGFR, PDGFR, and KIT. It has shown efficacy in mouse-model of PCa bone metastasis, and is presently in clinical trials for several cancers. We observed that both androgen receptor (AR) positive and AR-negative PCa cells differentiate into a NE phenotype upon treatment with Dovitinib. The NE differentiation was also observed when mice harboring PC3-xenografted tumors were systemically treated with Dovitinib. The mechanistic underpinnings of this differentiation are unclear, but seem to be supported through MAPK-, PI3K-, and Wnt-signaling pathways. Further elucidation of the differentiation process will enable the identification of alternative salvage or combination therapies to overcome the potential resistance development.     

Terpenoids. III: Synthesis and biological evaluation of 23-hydroxybetulinic acid derivatives as novel inhibitors of glycogen phosphorylase

A series of 23-hydroxybetulinic acid derivatives were prepared and tested in vitro as a new class of inhibitors of glycogen phosphorylase (GP). Within this series of compounds, 12b (IC₅₀ = 3.5 μM) is the most potent GPa inhibitor. The preliminary SAR results of the 23-hydroxybetulinic acid derivatives are discussed.     

PEX慢病毒载体构建及其在293FT细胞中的分泌表达

目的 构建含人MMP-9信号肽-MMP-2-PEX片段的重组慢病毒,并在293FT细胞中分泌表达PEX蛋白.方法 利用RT-PCR、基因重组等技术,构建含MMP-9信号肽-MMP-2-PEX片段的重组慢病毒表达载体pBPLV-signal-PEX,在脂质体介导下与包装质粒（pLP1、pLP2）、包膜质粒（pLP/VSVG）共转染293FT细胞,包装产生慢病毒并进行滴度测定.慢病毒感染2931;3＂细胞后,Western印迹法检测293FI＂细胞培养上清中PEX的表达.结果 酶切和DNA测序表明慢病毒表达载pBPLV-signal-PEX构建正确,四质粒共转染293FT细胞成功获得慢病毒;慢病毒感染293FT细胞后,在细胞培养卜清中可检测到PEX蛋白的表达.结论 成功构建了含人PEX的重组慢病毒,在体外有效感染293fT细胞并持续分泌表达目的 蛋白,为进一步研究PEX在肿瘤侵袭与转移中的作用提供实验基础.     

Conjugation of nattokinase and lumbrukinase with magnetic nanoparticles for the assay of their thrombolytic activities

Two important thrombolytic enzymes, nattokinase (NK) and lumbrukinase (LK), were immobilized onto fine magnetic Fe₃O₄ nanoparticles using 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide (EDC) as the coupling reagent, and their thrombolytic activities were studied. The Fe₃O₄ nanoparticles and NK- and LK-conjugated magnetic nanoparticles were characterized by transmission electron microscopy, Fourier transform infrared spectrophotometry, vibrating sample magnetometry, X-ray diffraction, and UV–vis absorption spectroscopy. Dual kinetic absorbance measurements at 405 and 630 nm were employed to measure their thrombolytic activity. Analysis of protein amount showed that the optimum conditions for NK and LK binding to nanoparticles were respectively at a mass ratio of 2:1:1, 2:1:2 (magnetic nanoparticles:protein:EDC), and pH 6.00. Thrombolytic activity assay showed that the best thrombolytic activity could reach 91.89% for NK–nanoparticle conjugates and 207.74% for LK–nanoparticle conjugates, which are much higher than the pure enzymes (NK, 82.86%; LK, 106.57%).     

Comparison of various kinds of bone marrow stem cells for the repair of infarcted myocardium: single clonally purified non-hematopoietic mesenchymal stem cells serve as a superior source

A variety of adult stem cells have been used to transplant into the infarcted (MI) heart, however, comparative studies are lacking to show more suitable source of cells for transplantation. We have identified a single non-hematopoietic mesenchymal stem cell subpopulation (snMSCs) isolated from human bone marrow and clonally purified, that over 99% of them expressed MSC marker proteins and cardiomyocyte marker proteins when induction in vitro. We also compared the effects of the snMSCs with unpurified MSC (uMSCs), mononuclear cells (BMMNCs), or peripheral blood mononuclear cells (PBMNCs) on myocardial repair after induction of MI in rats. Ninety days later, we observed a better cardiac function assessed by ejection fraction, fraction of shortening and lung wet/dry weight ratios, less remodeling of left ventricle (LV), lower collagen density in the LV, and more vessels in the ischemic wall in the snMSCs transplantation group than in other cell-transplanted groups. Furthermore, the transplanted cells expressing cardiomyocyte specific proteins or vascular endothelial cell marker proteins were more in the snMSCs group than in other ones. We conclude that transplantation with single clonally purified MSCs seems to be more beneficial to the cardiac repair than with other stem cells after MI.     

Synthesis and biological evaluation of novel 1-O- and 14-O-derivatives of oridonin as potential anticancer drug candidates

Novel 1-O- and 14-O-derivatives of oridonin were synthesized and biologically evaluated. All of the derivatives exhibited stronger cytotoxicity against six cancer cell lines (BGC-7901, SW-480, HL-60, BEL-7402, A549, and B16) than oridonin in vitro, and some of them were more potent than oridonin and cyclophosphamide in vivo. Compounds Ib and IIg were the most potent with the IC(50) values of 0.84 microM for Ib in HL-60 cell and 1.00 microM for IIg in BEL-7402 cell.     

Mammalian Diaphanous-Related Formin 1 Regulates GSK3β-Dependent Microtubule Dynamics Required for T Cell Migratory Polarization

The mammalian diaphanous-related formin (mDia1), a Rho-regulated cytoskeletal modulator, has been shown to promote T lymphocyte chemotaxis and interaction with antigen presenting cells, but the mechanisms underpinning mDia1 roles in these processes have not been defined. Here we show that mDia1^-/- T cells exhibit impaired lymphocyte function-associated antigen 1 (LFA-1)-mediated T cell adhesion, migration and in vivo trafficking. These defects are associated with impaired microtubule (MT) polarization and stabilization, altered MT dynamics and reduced peripheral clustering of the MT plus-end-protein, adenomatous polyposis coli (APC) in migrating T cells following LFA-1-engagement. Loss of mDia1 also leads to impaired inducible inactivation of the glycogen synthase kinase (GSK) 3β as well as hyperphosphorylation and reduced levels of APC in migrating T cells. These findings identify essential roles for the mDia1 formin in modulating GSK3β-dependent MT contributions to induction of T-cell polarity, adhesion and motility.     

Design,synthesis, and biological evaluation of 1,2,4-triazole bearing 5-substituted biphenyl-2-sulfonamide derivatives as potential antihypertensive candidates

A series of novel 1,2,4-triazole bearing 5-substituted biphenyl-2-sulfonamide derivatives were designed and synthesized to develop new angiotensin II subtype 2 (AT₂) receptor agonists as novel antihypertensive candidates. It was found that 14f (IC₅₀ = 0.4 nM) and 15e (IC₅₀ = 5.0 nM) displayed potent AT₂ receptor affinity and selectivity in binding assays. Biological evaluation in vivo suggested that 14f is obviously superior to that of reference drug losartan in RHRs, and meanwhile, 14f has no significant impact on heart rate. The interesting activities of these compounds may make them promising candidates as antihypertensive agents.