Nanopore sensor for fast label-free detection of short double-stranded DNAs

Functionalizing surface enhanced the molecular sensing ability of a fabricated nanopore by increasing the translocation duration time for a short double-stranded DNA. The surface of nanopore was derivatized with γ-aminopropyltriethoxysilane and the positively charged surface attracted DNA molecules when they were in the vicinity of nanopore. The translocation duration time of DNA increased due to the strong electrostatic interaction and it enabled us to detect a short double-stranded DNA (<1 kbp) that is under the size limit of a conventional solid state nanopore sensor. Both 539 and 910 bp double-stranded DNAs were analyzed with the surface functionalized nanopore and their translocation kinetics are presented in this work. The new feature of the surface modified nanopore that can detect short double-stranded DNA molecules could readily be applied for a rapid label-free diagnostic analysis in a Lab-On-a-Chip type DNA sensor.     

SAPAP3 regulates epileptic seizures involving GluN2A in post-synaptic densities

Aberrantly synchronized neuronal discharges in the brain lead to epilepsy, a devastating neurological disease whose pathogenesis and mechanism are unclear. SAPAP3, a cytoskeletal protein expressed at high levels in the postsynaptic density (PSD) of excitatory synapses, has been well studied in the striatum, but the role of SAPAP3 in epilepsy remains elusive. In this study, we sought to investigate the molecular, cellular, electrophysiological and behavioral consequences of SAPAP3 perturbations in the mouse hippocampus. We identified a significant increase in the SAPAP3 levels in patients with temporal lobe epilepsy (TLE) and in mouse models of epilepsy. In addition, behavioral studies showed that the downregulation of SAPAP3 by shRNA decreased the seizure severity and that the overexpression of SAPAP3 by recombinant SAPAP3 yielded the opposite effect. Moreover, SAPAP3 affected action potentials (APs), miniature excitatory postsynaptic currents (mEPSCs) and N-methyl-D-aspartate receptor (NMDAR)-mediated currents in the CA1 region, which indicated that SAPAP3 plays an important role in excitatory synaptic transmission. Additionally, the levels of the GluN2A protein, which is involved in synaptic function, were perturbed in the hippocampal PSD, and this perturbation was accompanied by ultrastructural morphological changes. These results revealed a previously unknown function of SAPAP3 in epileptogenesis and showed that SAPAP3 may represent a novel target for the treatment of epilepsy.Subject terms: Diagnostic markers, Epilepsy     

A CRM1-dependent nuclear export signal controls nucleocytoplasmic translocation of HSCARG, which regulates NF-κB activity

HSCARG is a newly identified nuclear factor-κB (NF-κB) inhibitor that plays important roles in cell growth. Our previous study found that HSCARG could shuttle between the nucleus and cytoplasm by sensing the change in cellular redox states. To further investigate the mechanism of HSCARG translocation and its effect on the regulation of NF-κB activity, we identified a previously uncharacterized nuclear export signal (NES) at residues 272-278 of HSCARG that is required for its cytoplasmic translocation. This leucine-rich NES was found to be mediated by chromosome region maintenance 1. More importantly, accumulation of HSCARG in the nucleus occurred following a mutation in the NES or oxidative stress, which attenuated the inhibition of NF-κB by HSCARG. These results indicate that nucleocytoplasmic translocation of HSCARG plays an important role in fine-tuning NF-κB signaling.     

蓝色长盾金小蜂对橡副珠蜡蚧的控制作用

蓝色长盾金小蜂Scutellista caerulea Fonscolombe是橡胶树重要害虫橡副珠蜡蚧Parasaissetia nigra Nietner的一种外寄生性天敌。为了明确蓝色长盾金小蜂对橡副珠蜡蚧的控害潜能,为该寄生蜂的进一步利用提供数据支撑。在室内通过体视镜下解剖观察寄生蜂在寄主腹下是否产卵测定了不同温度、寄主发育阶段下蓝色长盾金小蜂的寄生功能反应、寻找效应和自身密度干扰效应。温度设置有21、24、27、30、33、36℃共6个处理,寄主发育阶段设初期成虫（1-2 d成虫）、褐色期成虫（体色褐色,产卵前3-4 d成虫）、黑色期成虫（体色黑色,产卵1-2 d成虫）三个处理。结果显示：蓝色长盾金小蜂对橡副珠蜡蚧的寄生功能反应符合Holling-Ⅱ型和Holling-Ⅲ型模型,在21-36℃范围内,33℃时该蜂寄生效能最大,为44.4201,21℃时最小,为9.2458;在橡副珠蜡蚧为初期成虫-黑色期成虫范围内,寄生效能由大到小为黑色期成虫>褐色期成虫>初期成虫,分别为18.9044、13.7410、7.2002。采用Hassell-Varley干扰模型对不同温度下蓝色长盾金小蜂受自身密度干扰的寄生作用率进行拟合,表明蓝色长盾金小蜂在寄生时存在种群内个体间自我干扰情况。温度会影响蓝色长盾金小蜂的搜寻和自我干扰,在21-33℃范围内,33℃搜寻常数和干扰常数达到最大值,分别为0.6116、0.7535。当温度为33℃,寄主发育阶段为黑色期成虫时,蓝色长盾金小蜂对橡副珠蜡蚧有较强的控制能力,自身干扰作用最强。     

Development and application of a T7 RNA polymerase-dependent expression system for antibiotic production improvement in <Emphasis Type="Italic">Streptomyces</Emphasis>

Objective

To develop a reliable and easy to use expression system for antibiotic production improvement of Streptomyces.

Results

A two-compound T7 RNA polymerase-dependent gene expression system was developed to fulfill this demand. In this system, the T7 RNA polymerase coding sequence was optimized based on the codon usage of Streptomyces coelicolor. To evaluate the functionality of this system, we constructed an activator gene overexpression strain for enhancement of actinorhodin production. By overexpression of the positive regulator actII-ORF4 with this system, the maximum actinorhodin yield of engineered strain was 15-fold higher and the fermentation time was decreased by 48 h.

Conclusion

The modified two-compound T7 expression system improves both antibiotic production and accelerates the fermentation process in Streptomyces. This provides a general and useful strategy for strain improvement of important antibiotic producing Streptomyces strains.

     

Silencing of circHIPK3 hampers platelet-derived growth factor-induced proliferation and migration in airway smooth muscle cells through the miR-375/MMP-16 axis

Cytotechnology - Emerging evidence has suggested a pivotal role of circular RNAs (circRNAs) in the progression of asthma. In this paper, we explored the mechanisms underlying the modulation of...     

截头堆砂白蚁新群体的形成及发展

本文主要论述在广州室温及恒温27℃、相对湿度80%条件下,截头堆砂白蚁Cryptotermes domesticus(Haviland)新群体的形成、繁殖特性及其群体的发展、发育状况.(1)在广州6～7月室温下,截头堆砂白蚁原始繁殖蚁自行配对后,产卵前期7～16天,平均10.33±2.32天,卵期46～71天,平均55.13±6.42天,巢龄为1年、2年的新建群体,子代个体分别为3～8头、10～16头;3年巢龄群体,子代个体12～35头;4年巢龄群体,兵蚁开始出现,子代个体23～57头,其中兵蚁1～3头.该种白蚁新建群体经历7年才发育成熟,此时群体子代个体36～115头,其中兵蚁1～4头.(2)在恒温27℃、相对湿度80%条件下,截头堆砂白蚁原始繁殖蚁配对后产卵前期8～18天,平均11.6±2.66天,卵期50～73天,平均57.8±5.79天.1年巢龄群体,子代个体6～10头;2年巢龄群体,兵蚁出现,子代个体16～34头,其中兵蚁1～2头.新建群体经历2～3年发育成熟,此时群体内的子代个体18～44头,其中兵蚁1～2头.     

落叶松体胚发育中5个miRNA前体与成熟体的表达

利用同源比对或RACE克隆了5个落叶松(Larix leptolepis) miRNA前体。结果显示, 在各物种miRNA前体间, 成熟序列高度相似, 但其它序列相似度差异大, 序列相似度与亲缘关系有关。采用qRT-PCR分析了5个miRNA、前体和靶基因在落叶松体胚8个发育阶段的表达变化。结果显示, miRNA表达最高峰出现在后期子叶胚, 暗示与促进胚胎休眠有关; 表达次高峰出现时期不同, 表现为miR397和miR408在PEMIII, miR398在早期单胚, miR156和miR166在早期子叶胚, 表明其与保持薄细胞壁、质子传递、顶端分生组织形成等调控有关。miRNA成熟体表达与前体含量不呈线性相关, 可能受多重调控。研究结果对于阐明MIR基因进化、表达调控及在体胚发育中的调控功能具有重要理论意义。