TSG101 promotes the proliferation,migration and invasion of hepatocellular carcinoma cells by regulating the PEG10

The tumour susceptibility gene 101 (TSG101) is reported to play important roles in the development and progression of several human cancers. However, its potential roles and underlined mechanisms in human hepatocellular carcinoma (HCC) are still needed to be further clarified. In the present study, we reported that knock down of TSG101 suppressed the proliferation, migration and invasion of HCC cells, while overexpression of TSG101 facilitated them. Molecularly, the results revealed that knock down of TSG101 significantly decreased the cell cycle related regulatory factor p53 and p21. In another point, knock down of TSG101 also obviously decreased the level of metallopeptidase inhibitor TIMP1 (Tissue inhibitors of metalloproteinases 1), which results in inhibition of MMP2, MMP7 and MMP9. In contrast, overexpression of TSG101 had opposite effects. The iTRAQ proteomics analysis identified that oncogenic protein PEG10 (Paternally expressed gene 10) might be a potential downstream target of TSG101. Further investigation showed that TSG101 interacted with PEG10 and protected it from proteasomal degradation thereby regulating the expression of p53, p21 and MMPs. Finally, we found that both TSG101 and PEG10 proteins are up‐regulated and presented a direct correlation in HCC patients. In conclusion, these results suggest that TSG101 is up‐regulated in human HCC patients, which may accelerate the proliferation, migration and invasion of HCC cells through regulating PEG10.     

New insights into the binding mode of pyridine-3-carboxamide inhibitors of E. coli DNA gyrase

Previously we have reported on a series of pyridine-3-carboxamide inhibitors of DNA gyrase and DNA topoisomerase IV that were designed using a computational de novo design approach and which showed promising antibacterial properties. Herein we describe the synthesis of additional examples from this series aimed specifically at DNA gyrase, along with crystal structures confirming the predicted mode of binding and in vitro ADME data which describe the drug-likeness of these compounds.     

蓝氏贾第虫无义mRNA的降解

无义介导的mRNA降解途径（nonsense-mediated mRNA decay,NMD）作为细胞内的一种重要的mRNA质量监控机制,可以降解含有提前终止密码子（premature termination codon,PTC）的异常转录本,从而避免截短蛋白质对细胞的毒害,但其详细的分子机制有待进一步阐释。蓝氏贾第虫（Giardia lamblia)作为一种寄生性单细胞原生动物,进化地位特殊,对其NMD途径的研究有利于阐明基因表达调控的分子和进化机制。本研究通过酵母双杂交及体外pull-down实验分析了贾第虫NMD途径因子上游移码蛋白1(Giardia lamblia up-frameshift 1,GlUPF1)、贾第虫RNA结合蛋白（Giardia lamblia HRP1, GlHRP1）、贾第虫核糖核酸外切酶（Giardia lamblia Ski7p,GlSki7p、Giardia lamblia XRN1,GlXRN1）之间的相互作用关系。结果表明,GlUPF1全长与GlHRP1、GlXRN1(1~500 aa)、GlSki7p间均可发生相互作用。而且GlUPF1的CH结构域和C端结构域分别与GlHRP1、GlXRN1（1~500 aa）、GlSki7p相互作用。说明GlUPF1在贾第虫NMD途径中作为招募平台,在无义mRNA识别和降解过程中发挥重要作用。为此,结合本实验室之前的研究结果,我们提出原生动物贾第虫的NMD途径：在提前终止密码子处SURF(SMG1-UPF1-eRF1-eRF3)复合物形成后,GlUPF1被磷脂酰肌醇3-激酶(suppressor with morphogenetic effect on genitalia 1,SMG1)磷酸化修饰, NMD途径激活,随后GlUPF1与HRP1相互作用,将转录本标记为NMD底物;GlUPF1进而招募下游贾第虫5′-3′核糖核酸降解酶GlXRN1、贾第虫3′-5′ 核糖核酸降解因子GlSki7p,最终降解靶标mRNA。     

PEG模拟干旱条件下木麻黄幼苗的生理变化

以木麻黄Casuarina equisetifolia15个家系品种为材料,利用聚乙二醇6000(PEG-6000)模拟水分胁迫环境,设置三个胁迫强度处理(轻度、中度、重度胁迫),研究受胁迫2月龄木麻黄幼苗丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、脯氨酸含量的变化。结果表明,经干旱胁迫处理后,各家系幼苗叶片MDA含量随胁迫强度增大而增大,但增幅差异较大;不同干旱胁迫水平下,各家系SOD活性存在一定差异;重度胁迫条件下,除粤杂交家系外,其余家系脯氨酸含量与对照相比均为显著增加。采用主成分分析法进行抗旱性综合评价,表明MDA含量是反映水分胁迫最重要的指标。各指标综合分析结果表明,家系701、莆20、抗3、惠2、南山7和东山2抗旱性能力较好;东山短、601、59、A13抗旱能力一般;千头、粤杂交、抗1、龙4和平5抗旱能力较差。     

适合免疫治疗的大鼠抗肾小球基底膜肾炎模型的建立

以线性表位肽P14（a3_127-148）作为抗原建立适合免疫治疗的抗肾小球基底膜（anti-glomerular basement membrane,GBM）病大鼠模型。采用大鼠后脚垫三点注射P14（a3_127-148）与弗氏佐剂乳化物的方法进行单次免疫,免疫前后每周采集24 h尿样和血样,所有大鼠在免疫后7 w处死。大鼠免疫后,肾炎模型组在各时间点的24 h尿蛋白、尿蛋白肌酐比值（albumin/urine creatinine ratio,ACR）、血肌酐及血尿素氮均明显高于对照组（P<0.05）;循环抗P14（a3_127-148）IgG抗体水平明显高于对照组(P<0.001);PAS染色可见节段性纤维素样坏死,富于细胞型新月体;免疫组化染色可见肾小球有明显的中性粒细胞和巨噬细胞浸润;免疫荧光检测可见IgG沿GBM呈强线性沉积;电镜观察到GBM的断裂和收缩;而对照组均未见改变。HE染色在所有大鼠中均未观察到肺部病变。使用P14（a3_127-148）线性肽免疫WKY大鼠成功建立了大鼠抗GBM病模型,有助于开发更为特异的免疫疗法。     

Escherichia coli AspP activity is enhanced by macromolecular crowding and by both glucose-1,6-bisphosphate and nucleotide-sugars

Escherichia coli ADP-sugar pyrophosphatase (AspP) is a "Nudix" hydrolase that catalyzes the hydrolytic breakdown of ADP-glucose linked to glycogen biosynthesis. Moderate increases of AspP activity in the cell are accompanied by significant reductions of the glycogen content. In vitro analyses showed that AspP activity is strongly enhanced by macromolecular crowding and by both glucose-1,6-bisphosphate and nucleotide-sugars, providing a first set of indicative evidences that AspP is a highly regulated enzyme. To our knowledge, AspP is the sole bacterial enzyme described to date which is activated by both G1,6P(2) and nucleotide-sugars.     

Asymmetry of syringomycin E channel studied by polymer partitioning

To probe the size of the ion channel formed by Pseudomonas syringae lipodepsipeptide syringomycin E, we use the partial blockage of ion current by penetrating poly(ethylene glycol)s. Earlier experiments with symmetric application of these polymers yielded a radius estimate of approximately 1 nm. Now, motivated by the asymmetric non-ohmic current-voltage curves reported for this channel, we explore its structural asymmetry. We gauge this asymmetry by studying the channel conductance after one-sided addition of differently sized poly(ethylene glycol)s. We find that small polymers added to the cis-side of the membrane (the side of lipodepsipeptide addition) reduce channel conductance much less than do the same polymers added to the trans-side. We interpret our results to suggest that the water-filled pore of the channel is conical with cis- and trans-radii differing by a factor of 2-3 and that the smaller cis-radius is in the 0.25-0.35 nm range. In symmetric, two-sided addition, polymers entering the pore from the larger opening dominate blockage.     

牛磺酸对高糖培养下视网膜Mü ller细胞GFAP和TAUT表达的影响

目的：通过检测高糖培养条件下视网膜Mü ller细胞神经纤维酸性蛋白（glial fibrillary acid protein,GFAP）和牛磺酸转运蛋白（taurine transporter,TAUT）的表达变化,观察葡萄糖对Mü ller细胞牛磺酸（taurine）转运功能的影响,探讨牛磺酸对早期糖尿病视网膜病（DR）可能的保护作用.方法：高糖培养大鼠视网膜Mü ller细胞,用免疫细胞荧光化学双染色、Western blotting技术检测不同浓度牛磺酸干预下Mü ller细胞GFAP及TAUT的蛋白表达.结果：高糖可引起Mü ller细胞GFAP表达增强,TAUT表达减弱;牛磺酸可减弱高糖引起的Mü ller细胞GFAP表达增强,TAUT在0.1mmol/L～10 mmo1/L的牛磺酸干预后表达增强.结论：牛磺酸可以抑制高糖导致的Müller细胞功能改变.     

海金沙卵发生的组织化学研究

采用组织化学方法对海金沙（Lygodium japonicum （Thunb.） Sw.）卵发生进行了研究。高碘酸-锡夫反应（PAS反应）结果显示：颈沟和腹沟细胞外产生的黏性物质为多糖类物质;在卵细胞发生早期颈卵器细胞内均含有质体,且淀粉粒含量丰富,随着颈卵器的发育,卵细胞以及颈沟和腹沟细胞内淀粉粒数量和体积均逐渐减少,最后退化消失。苏丹黑B反应结果显示：海金沙颈沟和腹沟细胞外产生的黏性物质也含有脂类物质,而卵细胞内并未有明显脂类物质产生;卵细胞质膜处也并没有糖类或脂类物质的积累。本研究从组织化学角度佐证了海金沙的特殊性,即既有原始蕨类的特征,又有进化蕨类的特征。     

Recent advances in plasmid-based tools for establishing novel microbial chassis

A key challenge for domesticating alternative cultivable microorganisms with biotechnological potential lies in the development of innovative technologies. Within this framework, a myriad of genetic tools has flourished, allowing the design and manipulation of complex synthetic circuits and genomes to become the general rule in many laboratories rather than the exception. More recently, with the development of novel technologies such as DNA automated synthesis/sequencing and powerful computational tools, molecular biology has entered the synthetic biology era. In the beginning, most of these technologies were established in traditional microbial models (known as chassis in the synthetic biology framework) such as Escherichia coli and Saccharomyces cerevisiae, enabling fast advances in the field and the validation of fundamental proofs of concept. However, it soon became clear that these organisms, although extremely useful for prototyping many genetic tools, were not ideal for a wide range of biotechnological tasks due to intrinsic limitations in their molecular/physiological properties. Over the last decade, researchers have been facing the great challenge of shifting from these model systems to non-conventional chassis with endogenous capacities for dealing with specific tasks. The key to address these issues includes the generation of narrow and broad host plasmid-based molecular tools and the development of novel methods for engineering genomes through homologous recombination systems, CRISPR/Cas9 and other alternative methods. Here, we address the most recent advances in plasmid-based tools for the construction of novel cell factories, including a guide for helping with “build-your-own” microbial host.