Electric Field-controlled Directed Migration of Neural Progenitor Cells in 2D and 3D Environments

Endogenous electric fields (EFs) occur naturally in vivo and play a critical role during tissue/organ development and regeneration, including that of the central nervous system^1,2. These endogenous EFs are generated by cellular regulation of ionic transport combined with the electrical resistance of cells and tissues. It has been reported that applied EF treatment can promote functional repair of spinal cord injuries in animals and humans^3,4. In particular, EF-directed cell migration has been demonstrated in a wide variety of cell types^5,6, including neural progenitor cells (NPCs)^7,8. Application of direct current (DC) EFs is not a commonly available technique in most laboratories. We have described detailed protocols for the application of DC EFs to cell and tissue cultures previously^5,11. Here we present a video demonstration of standard methods based on a calculated field strength to set up 2D and 3D environments for NPCs, and to investigate cellular responses to EF stimulation in both single cell growth conditions in 2D, and the organotypic spinal cord slice in 3D. The spinal cordslice is an ideal recipient tissue for studying NPC ex vivo behaviours, post-transplantation, because the cytoarchitectonic tissue organization is well preserved within these cultures^9,10. Additionally, this ex vivo model also allows procedures that are not technically feasible to track cells in vivo using time-lapse recording at the single cell level. It is critically essential to evaluate cell behaviours in not only a 2D environment, but also in a 3D organotypic condition which mimicks the in vivo environment. This system will allow high-resolution imaging using cover glass-based dishes in tissue or organ culture with 3D tracking of single cell migration in vitro and ex vivo and can be an intermediate step before moving onto in vivo paradigms.     

西藏蓝钟花属一新变型——白花裂叶蓝钟花

描述了桔梗科蓝钟花属裂叶蓝钟花(Cyananthus lobatus Wall.ex Benth.)的一个新变型——白花裂叶蓝钟花(Cyananthus lobatus Wall.ex Benth.f.albiflorus J.Luo et S.L.Wang)。原变型的花冠为紫蓝色至淡蓝色,而新变型花冠为白色。     

石蒜儿茶酚氧位甲基转移酶基因克隆与原核表达

采用同源克隆与RACE相结合的方法,首次从石蒜叶片中克隆到可能与加兰他敏生物合成相关的儿茶酚氧位甲基转移酶基因,命名为LrCOMT。核酸序列分析显示,该cDNA全长1 246bp,开放阅读框1 101bp,编码366氨基酸。氨基酸序列分析与比对发现,LrCOMT编码的氨基酸序列具有COMT蛋白的特征区域,且与鸢尾、玉米、烟草等植物COMT的同源性大于60%。将LrCOMT基因连接到原核表达载体pET-29a上,转化大肠杆菌BL21(DE3)的原核表达结果显示,重组蛋白受IPTG诱导表达,分子量大小约为40kD,与已报道的其他植物COMT大小基本一致。     

异地半圈养藏羚卧息行为适应性的初步研究

藏羚Pantholops hodgsoni是青藏高原特有物种。为了探讨藏羚羊对异地环境的行为学适应,对异地圈养藏羚羊的卧息行为进行了不同性别、不同季节间的比较研究。采用全事件记录法和焦点动物取样法,记录和统计了异地圈养藏羚羊冷季和暖季的卧息行为。研究结果表明,在暖季,雌性和雄性藏羚羊在卧息时间及卧息时间占全天活动时间的比例(卧息比例)上没有差异(卧息时间:Z=-1.126,P0.05;卧息比例:Z=-1.357,P0.05);在冷季,雌性和雄性藏羚羊在卧息时间及卧息比例上也没有差异(卧息时间:Z=-1.14,P0.05;卧息比例:Z=-0.307,P0.05)。雄、雌性藏羚羊的卧息时间及卧息比例没有季节差异(雄性卧息时间:Z=-1.465,P0.05;雄性卧息比例:Z=-1.631,P0.05;雌性卧息时间:Z=-1.742,P0.05;雌性卧息比例:Z=-1.852,P0.05)。季节和性别对藏羚羊卧息行为的影响没有交互作用(F=0.057,P0.05)。总之,藏羚羊卧息行为之间既没有性别差异也没有季节差异,与可可西里地区藏羚羊的卧息行为相比,异地圈养藏羚羊的卧息行为已经调整其行为学适应对策。     

Therapeutic response to CDK4/6 inhibition in breast cancer defined by ex vivo analyses of human tumors

To model the heterogeneity of breast cancer as observed in the clinic, we employed an ex vivo model of breast tumor tissue. This methodology maintained the histological integrity of the tumor tissue in unselected breast cancers, and importantly, the explants retained key molecular markers that are currently used to guide breast cancer treatment (e.g., ER and Her2 status). The primary tumors displayed the expected wide range of positivity for the proliferation marker Ki67, and a strong positive correlation between the Ki67 indices of the primary and corresponding explanted tumor tissues was observed. Collectively, these findings indicate that multiple facets of tumor pathophysiology are recapitulated in this ex vivo model. To interrogate the potential of this preclinical model to inform determinants of therapeutic response, we investigated the cytostatic response to the CDK4/6 inhibitor, PD-0332991. This inhibitor was highly effective at suppressing proliferation in approximately 85% of cases, irrespective of ER or HER2 status. However, 15% of cases were completely resistant to PD-0332991. Marker analyses in both the primary tumor tissue and the corresponding explant revealed that cases resistant to CDK4/6 inhibition lacked the RB-tumor suppressor. These studies provide important insights into the spectrum of breast tumors that could be treated with CDK4/6 inhibitors, and defines functional determinants of response analogous to those identified through neoadjuvant studies.     

拟南芥新型脂转移蛋白AtDHyPRP1的亚细胞定位及其对灰霉菌的抗性

通过遗传转化技术研究了拟南芥脂转移蛋白AtDHyPRP1在细胞中的定位及其对真菌病原体的抗性。采用PCR方法从拟南芥Ws生态型克隆了AtDHyPRP1基因,构建产生pRI101-AN-AtDHyPRP1植物双元表达载体和pCAMBIA1302-AtDHyPRP1-GFP融合表达载体,经农杆菌介导的叶盘和浸花法得到烟草和拟南芥转基因植株。AtDHyPRP1基因能够明显增加烟草对灰霉菌的抗性,转AtDHyPRP1烟草叶片的被侵染部位有大量H2O2积累,激光共聚焦显微观察发现AtDHyPRP1蛋白定位于细胞表面。说明AtDHyPRP1蛋白在合成后被分泌到细胞外执行特殊的功能,与植物抗病防御机制有关。     

云南药用野生稻BIBAC文库混合克隆池制备及筛选

在已构建完成的云南药用野生稻BIBAC( binary bacterial artificial chomosome)文库的基础上,将文库制备成一、二、三级混合克隆池,各级混合池的数量分别为3 360、140和14个.根据Xa21抗病基因序列设计1对特异引物,利用4步PCR法对文库混合克隆池进行逐级筛选,初步确定了3个抗病基因阳性克隆.为今后以PCR法高效利用云南药用野生稻BIBAC文库挖掘其优异基因奠定了良好的基础.     

中国竹亚科矢竹属一新种——中岩茶秆竹

描述了竹亚科矢竹属一新种——中岩茶秆竹（Pseudosasa zhongyanensis S.H.Chen,K.F.Huang et H.Z.Guo）。本种与茶秆竹（P.amabilis(McClure) Keng f.）相近似,但箨环具明显箨基残留物和木栓质隆起,高于秆环,箨鞘顶端具小型外翻箨耳,耳缘具皱曲繸毛,箨舌几无毛或有短柔毛,易于区别。     

毛头鬼伞多糖CCP60a对TMV外壳蛋白的影响

对毛头鬼伞（Coprinus comatus Muell．ex Fr．）子实体中的多糖CCP60a进行了提取分离及检测,研究了不同温度条件下CCP60a对烟草花叶病毒（TMV）外壳蛋白体外聚合的影响,并用Western blotting法研究了CCP60a对TMV外壳蛋白表达的影响。结果表明,经沸水浸提、乙醇分级沉淀、DEAESephadex A-25离子交换柱层析、Sepharose-6B凝胶柱层析和抗TMV活性跟踪检测可得到均一的多糖CCP60a。随温度的提高,CCP60a处理组在320nm处的吸光度值增加幅度明显小于对照组,表明CCP60a对TMV外壳蛋白体外聚合有一定的抑制作用。Western blotting检测结果显示,CCP60a可以使TMV外壳蛋白的表达明显降低。     

石蒜生长对杂草群落组成及物种多样性的影响

通过野外调查,研究了石蒜（Lycoris radiata）对群落杂草组成和物种多样性的影响。结果表明：石蒜的生长能减少群落中物种的种类,降低群落中杂草的密度,并减少商陆（Phytolacca aeinosa）、蹄盖蕨（Athyrium fricellia）及禾本科（Granmeae）几个物种的数量,群落的物种多样性指数降低,群落均匀度提高。群落相似度与没有石蒜分布的群落相比显著降低。说明石蒜改变了杂草的群落结构,有利于限制杂草的发生危害。