Ultrasound-Triggered Phase Transition Sensitive Magnetic Fluorescent Nanodroplets as a Multimodal Imaging Contrast Agent in Rat and Mouse Model

Ultrasound-triggered phase transition sensitive nanodroplets with multimodal imaging functionality were prepared via premix Shirasu porous glass (SPG) membrane emulsification method. The nanodroplets with fluorescence dye DiR and SPIO nanoparticles (DiR-SPIO-NDs) had a polymer shell and a liquid perfluoropentane (PFP) core. The as-formed DiR-SPIO-NDs have a uniform size of 385±5.0 nm with PDI of 0.169±0.011. The TEM and microscopy imaging showed that the DiR-SPIO-NDs existed as core-shell spheres, and DiR and SPIO nanoparticles dispersed in the shell or core. The MTT and hemolysis studies demonstrated that the nanodroplets were biocompatible and safe. Moreover, the proposed nanodroplets exhibited significant ultrasound-triggered phase transition property under clinical diagnostic ultrasound irradiation due to the vaporization of PFP inside. Meanwhile, the high stability and R₂ relaxivity of the DiR-SPIO-NDs suggested its applicability in MRI. The in vivo T₂-weighted images of MRI and fluorescence images both showed that the image contrast in liver and spleen of rats and mice model were enhanced after the intravenous injection of DiR-SPIO-NDs. Furthermore, the ultrasound imaging (US) in mice tumor as well as MRI and fluorescence imaging in liver of rats and mice showed that the DiR-SPIO-NDs had long-lasting contrast ability in vivo. These in vitro and in vivo findings suggested that DiR-SPIO-NDs could potentially be a great MRI/US/fluorescence multimodal imaging contrast agent in the diagnosis of liver tissue diseases.     

沙地云杉林研究述评

沙地云杉(Picea mongolica)是我国特有的针叶林树种,主要分布于内蒙古白音敖包国家级自然保护区。在该地区的防风治沙、退化土壤的生态恢复,三北防护林体系建设等方面起着重要作用。从沙地云杉林的生长环境、个体生态、种群生态、群落生态等方面对国内外的研究现状和最新研究进展进行了综述,并从不同尺度上对沙地云杉林未来研究中应该重视的一些科学问题进行了展望。这些科学问题主要有:(1)沙地云杉个体生长、生殖和保护的结构和生理机理方面的研究。(2)沙地云杉种群与群落方面的研究包括:生命表与生育力表编制与统计;种内与种间的关系;化感作用;种群、群落发生、发展和衰亡的内在机制;自然干扰和人为干扰对沙地云杉更新的研究;全球气温变化对沙地云杉林天然更新的影响等。(3)沙地云杉林生态系统方面的研究包括:大量和微量营养元素在沙地云杉林的凋落物⁃微生物⁃土壤⁃沙地云杉的循环、利用规律及转化机理;凋落物分解规律及机理;水分循环;土壤理化性质和生化特性对沙地云杉生长和生物量的影响;沙地云杉林生态系统能量流动和信息传递研究等。(4)沙地云杉林生态经营研究;沙地云杉丰产林营造技术;沙地云杉种子园、母树林规划与创建;沙地云杉林杄插繁殖技术与无性系研究;幼树移栽技术;沙地云杉天然林和人工林施肥技术;沙地云杉林病虫害预测与防治研究等。     

利用皆伐法估算黔中喀斯特森林地上生物量

精确估算森林生物量对理解全球碳循环至关重要。已有的喀斯特森林地上生物量估算研究存在很高的不确定性,缺乏校准数据检验研究结果的精度。利用皆伐法,首次精确估算了我国西南贵州省中部喀斯特森林的地上生物量,并检验了已有生物量回归方程和平均标准木法对该喀斯特森林地上生物量的估算效果。该喀斯特森林的地上生物量为122.81 Mg/hm~2,胸径(D)≥1 cm的木本植物、D1 cm的木本植物和草本植物的地上生物量分别为120.00、2.56、0.24 Mg/hm~2。D在10—30 cm范围内的植株(83.89 Mg/hm~2)是地上生物量的主要贡献者。4个优势树种(云南鼠刺Itea yunnanensis、川钓樟Lindera pulcherrima、猴樟Cinnamomum bodinieri和化香树Platycarya strobilacea的地上生物量为103.03 Mg/hm~2,占森林总地上生物量的83.89%。干(61.04 Mg/hm~2)和枝(40.56 Mg/hm~2)的生物量远高于皮(11.61 Mg/hm~2)和叶(6.80 Mg/hm~2)。在物种水平上,已有生物量回归方程(误差-56.10%—84.61%)和平均标准木法(误差-36.43%—-5.14%)对该喀斯特森林地上生物量的估算效果均较差。最后,建立了5个新的生物量回归方程。本研究可为我国西南喀斯特地区精确估算森林碳储量提供基础校验数据和方法指导。     

Collagen IV contributes to nitric oxide-induced angiogenesis of lung endothelial cells

Nitric oxide (NO) mediates endothelial angiogenesis via inducing the expression of integrin α(v)β(3). During angiogenesis, endothelial cells adhere to and migrate into the extracellular matrix through integrins. Collagen IV binds to integrin α(v)β(3), leading to integrin activation, which affects a number of signaling processes in endothelial cells. In the present study, we evaluated the role of collagen IV in NO-induced angiogenesis. We found that NO donor 2,2'-(hydroxynitrosohydrazino)bis-ethanamine (NOC-18) causes increases in collagen IV mRNA and protein in lung endothelial cells and collagen IV release into the medium. Addition of collagen IV into the coating of endothelial culture increases endothelial monolayer wound repair, proliferation, and tube formation. Inhibition of collagen IV synthesis using gene silencing attenuates NOC-18-induced increases in monolayer wound repair, cell proliferation, and tube formation as well as in the phosphorylation of focal adhesion kinase (FAK). Integrin blocking antibody LM609 prevents NOC-18-induced increase in endothelial monolayer wound repair. Inhibition of protein kinase G (PKG) using the specific PKG inhibitor KT5823 or PKG small interfering RNA prevents NOC-18-induced increases in collagen IV protein and mRNA and endothelial angiogenesis. Together, these results indicate that NO promotes collagen IV synthesis via a PKG signaling pathway and that the increase in collagen IV synthesis contributes to NO-induced angiogenesis of lung endothelial cells through integrin-FAK signaling. Manipulation of collagen IV could be a novel approach for the prevention and treatment of diseases such as alveolar capillary dysplasia, severe pulmonary arterial hypertension, and tumor invasion.     

Phosvitin plays a critical role in the immunity of zebrafish embryos via acting as a pattern recognition receptor and an antimicrobial effector

How fish embryos that develop externally survive microbial attacks is poorly understood. Here, we clearly demonstrated that the embryo extract of zebrafish and its early embryo both displayed antimicrobial activity against microbes, including pathogenic Aeromonas hydrophila, and phosvitin (Pv), a nutritional protein abundant in eggs, was related to this antimicrobial activity. We also showed that recombinant Pv (rPv) acted as a pattern recognition receptor capable of recognizing the microbial signature molecules LPS, lipoteichoic acid, and peptidoglycan, as well as binding the Gram-negative and -positive microbes Escherichia coli, A. hydrophila, and Staphylococcus aureus and functioned as an antimicrobial agent capable of killing the microbes. Furthermore, we revealed that its C-terminal 55 residues (Pt5) with the functional sites Arg(242) and Ala(201)/Ile(203) were indispensable for Pv antimicrobial activity. Importantly, microinjection of rPv or Pt5 into early embryos significantly enhanced their resistance to A. hydrophila challenge, and this enhanced bacterial resistance was markedly reduced by co-injection of anti-Pv antibody plus rPv (or Pt5) but not by injection of anti-actin antibody plus rPv. Moreover, the generated mutants with in vitro antimicrobial activity, when injected into the embryos, could also promote their resistance to A. hydrophila, but those without in vitro antimicrobial activity could not. It is thus proposed that Pv participates in the protection of early embryos against pathogenic attacks via binding and disrupting potential pathogens. This work also opens a new way for the study of the immunological roles of yolk proteins in oviparous animals that rely on yolk proteins for embryonic development.     

Inhibitory effect of Disulfiram/copper complex on non-small cell lung cancer cells

Non-small cell lung cancer (NSCLC) is the most common cause of cancer-related death in both men and women worldwide. Recently, Disulfiram has been reported to be able to inhibit glioblastoma, prostate, or breast cancer cell proliferation. In this study, the synergistic effect of Disulfiram and copper on NSCLC cell growth was investigated. Inhibition of cancer cell proliferation was detected by 1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT) assay and cell cycle analysis. Liquid colony formation and tumor spheroid formation assays were used to evaluate their effect on cancer cell clonogenicity. Real-time PCR was performed to test the mRNA level of cancer stem cell related genes. We found that Disulfiram or copper alone did not potently inhibit NSCLC cell proliferation in vitro. However, the presence of copper significantly enhanced inhibitory effect of Disulfiram on NSCLC cell growth, indicating a synergistic effect between Disulfiram and copper. Cell cycle analysis showed that Disulfiram/copper complex caused NSCLC cell cycle arrest in G2/M phase. Furthermore, Disulfiram/copper significantly increased the sensitivity of cisplatin in NSCLC cells tested by MTT assay. Liquid colony formation assay revealed that copper dramatically increased the inhibitory effect of Disulfiram on NSCLC cell colony forming ability. Disulfiram combined with copper significantly attenuated NSCLC cell spheroid formation and recuded the mRNA expression of lung cancer stem cell related genes. Our data suggest that Disulfiram/copper complex alone or combined with other chemotherapy is a potential therapeutic strategy for NSCLC patients.     

玉米AGO基因的克隆与表达分析

以玉米自交系‘昌7-2’三叶期前后2个时间点(种子萌发后5d和8d)幼苗不同组织部位的总RNA为研究对象,采用实时荧光定量PCR技术,对玉米中6个Argonaute(AGO)蛋白家族基因(AGO1、AGO2、AGO4、AGO5、AGO7和AGO10)在幼苗不同发育时期及不同组织部位的表达谱进行了研究。结果表明:(1)AGO1、AGO2、AGO4和AGO7在种子萌发后5d和8d幼苗不同组织中均有表达,种子萌发后5d幼苗中的平均表达量均高于萌发8d的幼苗,且在地上部分新生组织或细胞分裂比较旺盛的组织中表达较多,表明AGO1、AGO2、AGO4和AGO7可能在玉米幼苗发育早期的分生组织分裂生长中发挥调控作用。(2)AGO5和AGO10只在叶片和茎尖中表达,其他组织中不表达;其中AGO5主要集中在新生叶和种子萌发后8d的茎尖中,AGO10在玉米叶发育过程中可能存在着迁移的现象。     

Multiple functions of an odorant-binding protein in the mosquito Aedes aegypti

To prevent spreading of deadly diseases, populations of mosquitoes can be controlled by interfering with their chemical communication system. Odorant-binding proteins, recently shown to be required for olfaction, represent interesting targets for such purpose. Here we describe the ligand-binding properties and the unusual tissue expression of odorant-binding protein 22 from the repertoire of Aedes aegypti. Best ligands are molecules with two aromatic rings connected by a short rigid chain. The protein is expressed not only in sensory organs, such as the antennae and proboscis, but also in the male reproductive apparatus and transferred to the spermathecs of females. This suggests an additional function for this protein as pheromone carrier, analogously to vertebrates’ urinary and salivary proteins as well as some insect chemosensory proteins. Antiserum against odorant-binding protein 22 also stained the edges and sensilla of spiracles, indicating a third, still unknown, role for this protein.     

Efficient, chemoselective synthesis of immunomicelles using single-domain antibodies with a C-terminal thioester

Background  

Classical bioconjugation strategies for generating antibody-functionalized nanoparticles are non-specific and typically result in heterogeneous compounds that can be compromised in activity. Expression systems based on self-cleavable intein domains allow the generation of recombinant proteins with a C-terminal thioester, providing a unique handle for site-specific conjugation using native chemical ligation (NCL). However, current methods to generate antibody fragments with C-terminal thioesters require cumbersome refolding procedures, effectively preventing application of NCL for antibody-mediated targeting and molecular imaging.