长江经济带国土空间开发与保护路径优化

长江经济带是重大国家战略发展区域,优化其国土空间开发保护格局是实现生态文明建设的重要内容。研究基于“核心点-发展轴-战略区”的框架,通过国土空间利用强度表征指标及夜间灯光数据识别发展核心点,借助有序加权平均算法模拟多种决策愿景下的空间发展阻力面,运用最小累积阻力模型和电路理论模型识别空间发展轴线及其内部的开发保护战略区,定量构建不同决策偏好的空间格局优化方案,突破了传统研究中单一目标和定性分析的局限。结果表明：长江经济带国土空间发展核心点共63个,约占区域总面积的5%,呈现自东向西递减的空间分布特征;城镇扩张、协同发展和生态约束三类情景下发展轴线均为61条,所覆盖的县级单元分别占总数的36.6%、36.5和36.7%,呈现出“一横三纵”的格局特征;城镇扩张、协同发展和生态约束三类情景下开发战略区分别占轴线面积的4.5%、4.9%、4.8%,多分布于长江干流及其支流沿线。保护战略区分别占轴线面积的17.1%、13.9%、15.3%,主要分布于长江南岸区域。在此基础上形成的国土空间开发保护格局基础架构,能够为落实长江经济带“共抓大保护、不搞大开发”提供决策参考。     

Expression,purification, and characterization of recombinant lumbrokinase PI239 in Escherichia coli

Lumbrokinase (LK) is an important fibrinolytic enzyme derived from earthworms. It has been found that LK is composed of a group of isoenzymes. To construct and express the mature peptide of LK PI239 in Escherichia coli, we amplified and optimized the gene of LK which was then cloned into the prokaryotic expression vector pET-22b(?). The recombinant LK (rLK) protein was expressed as inclusion bodies and we have developed a purification process of rLK from these inclusion bodies. A step-down urea concentration strategy was applied to the rLK renaturation process. The purified and renatured rLK apparently ameliorated the conditions of the model thrombosis rats used, and may be developed into a therapeutic agent for thrombotic-associated diseases.     

Subcellular localization and functional characterization of a fish IRF9 from crucian carp Carassius auratus

Mammalian interferon (IFN) regulatory factor 9 (IRF-9) has long been recognized as the DNA sequence recognition subunit of IFN-stimulated gene factor 3 (ISGF3) complex, which is critical for type I IFN to induce the expression of IFN-stimulated genes (ISGs) against viral infection. Recent studies have shown that fish IFN exerts antiviral effects by induction of a number of ISGs and also of itself; however, little is known about the role of fish IRF9 in IFN signaling. Here we identify a fish IRF9 orthologue (CaIRF9) from IFN-producing cell line, crucian carp Carassius auratus blastulae embryonic (CAB) cells. Analysis of subcellular distribution of CaIRF9-green fluorescent protein indicates that CaIRF9 is constitutively present in the nucleus, which is driven by two nuclear localization signals (NLS), one locating within DNA-binding domain (DBD) of CaIRF9 and the other immediately behind DBD, although human IRF9 contains only one NLS analogous to the former of CaIRF9. Overexpression of CaIRF9 together with CaSTAT2 not only activates ISRE-containing promoter but also upregulates the expression of fish ISGs. Strikingly, CaIRF9 together with CaSTAT2 also exhibits an ability to activate crucian carp IFN promoter, and blockade of cellular CaIRF9 attenuates IFN itself-induced activation of crucian carp IFN promoter. Taken together, these data suggest that crucian carp IFN induces the expression of ISGs and also of itself possibly by the JAK-STAT signaling pathway that is conserved from fish to mammals.     

Interspecific delimitation and phylogenetic origin of Pugionium(Brassicaceae)

Pugionium(Brassicaceae)is a small genus that occurs in central Asian deserts.The interspecific delimitation and taxonomic treatments of this genus are disputed and its phylogenetic origin remains unknown. In the present study,we examined these issues based on morphological and molecular data obtained for the first time.We used statistical methods to examine inter-and intraspecific morphological variations.The results suggest that only two species,namely P.dolabratum and P.cornutum,can be warranted for all examined populations and specimens,whereas three species(P.calcaratum,P.cristatum,and P.pterocarpum)should be incorporated into P. dolabratum.This delimitation was further supported by the molecular data:all populations of P.dolabratum,P. calcaratum,P.cristatum,and P.pterocarpum shared the same internal transcribed spacer genotype,whereas those from P.cornutum had another type.Phylogenetic analyses of Pugionium and representative genera of Brassicaceae based on ndhF sequences suggest that this genus is sister to the genus Megacarpaea,which,together,comprise a well-supported lineage with Farsetia,Lobularia,Iberis,and Ionopsidium,whereas the two other genera that were previously suggested to be closely related to this genus(Isatis and Bunias)were placed in the other lineages.We further discuss the origin of Pugionium and suggest that it probably originated in central Asia when the climate became drier from the late Miocene.     

Synthetic miRNA-Mowers Targeting miR-183-96-182 Cluster or miR-210 Inhibit Growth and Migration and Induce Apoptosis in Bladder Cancer Cells

Background

MicroRNAs (miRNAs) function as endogenous regulators of biological behaviors of human cancers. Several natural non-coding RNAs are reported to inhibit miRNAs by base-pairing interactions. These phenomena raise questions about the ability of artificial device to regulate miRNAs. The purpose of this study is to create synthetic devices that target a single miRNA or a miRNA cluster and to ascertain their therapeutic effects on the phenotypes of bladder cancer cells.

Methodology/Principal Findings

Tandem bulged miRNA binding sites were inserted into the 3′ untranslated region (UTR) of the SV-40 promoter-driven Renilla luciferase gene to construct two “miRNA-mowers” for suppression of miR-183-96-182 cluster or miR-210. A third device with tandem repeat sequences not complementary to any known miRNA was generated as an untargeted-control. In functional analyses, bladder cancer T24 and UM-UC-3 cells were transfected with each of the three devices, followed by assays for detection of their impacts. Luciferase assays indicated that the activities of the luciferase reporters in the miRNA-mowers were decreased to 30–50% of the untargeted-control. Using Real-Time qPCR, the expression levels of the target miRNAs were shown to be reduced 2-3-fold by the corresponding miRNA-mower. Cell growth, apoptosis, and migration were tested by MTT assay, flow cytometry assay, and in vitro scratch assay, respectively. Cell growth inhibition, increased apoptosis, and decreased motility were observed in miRNA-mowers-transfected bladder cancer cells.

Conclusions/Significance

Not only a single target miRNA but also the whole members of a target miRNA cluster can be blocked using this modular design strategy. Anti-cancer effects are induced by the synthetic miRNA-mowers in the bladder cancer cell lines. miR-183/96/182 cluster and miR-210 are shown to play oncogenic roles in bladder cancer. A potentially useful synthetic biology platform for miRNA loss-of-function study and cancer treatment has been established in this work.     

结球甘蓝下胚轴原生质体培养再生植株体系的优化研究

以结球甘蓝‘新夏50’的无菌苗下胚轴为材料,对影响原生质体分离、纯化与培养的主要因素进行研究,建立适合结球甘蓝原生质体游离、纯化、收集、培养以至再生出完整植株的实用技术体系,为其非对称细胞融合及品种改良与创新等研究奠定基础。结果表明:2.5%纤维素酶R-10+0.05%果胶酶Y-23+9CPW+5mmol/L MES的混合酶液,从4d苗龄的下胚轴上分离出高产率的原生质体。在改良B5+0.5mg/L 2,4-D+0.2mg/L 6-BA+0.2mg/L NAA的液体培养基上,原生质体分裂旺盛。形成愈伤组织后经芽诱导和生根培养,获得了再生植株。倍性检测结果表明,不同原生质体所获得的24株再生植株中,19株为正常二倍体,4株为嵌合体,1株为四倍体。     

Trimethyloxonium modification of single batrachotoxin-activated sodium channels in planar bilayers. Changes in unit conductance and in block by saxitoxin and calcium

Single batrachotoxin-activated sodium channels from rat brain were modified by trimethyloxonium (TMO) after incorporation in planar lipid bilayers. TMO modification eliminated saxitoxin (STX) sensitivity, reduced the single channel conductance by 37%, and reduced calcium block of inward sodium currents. These effects always occurred concomitantly, in an all-or-none fashion. Calcium and STX protected sodium channels from TMO modification with potencies similar to their affinities for block. Calcium inhibited STX binding to rat brain membrane vesicles and relieved toxin block of channels in bilayers, apparently by competing with STX for the toxin binding site. These results suggest that toxins, permeant cations, and blocking cations can interact with a common site on the sodium channel near the extracellular surface. It is likely that permeant cations transiently bind to this superficial site, as the first of several steps in passing inward through the channel.