链霉菌分化调控启动子——PTH4和PTH270的体内转录研究

天蓝色链霉菌分化调控启动子P_TH4和P_TH270被分别亚克隆到链霉菌启动子探针载体pIJ4083后,构建的重组质粒被命名为pIJ4470和pIJ4471.当pIJ4470和pIJ4471转化天蓝色链霉菌的白色分化阻断突变株(C85,C70,C71,C17和C119)后,通过pIJ4083上儿茶酚加氧酶报告基因的表达可知启动子的活性.从构建的重组菌株中进行了总RNA的提取.用同位素标记P_TH4和P_TH270的5’-端制备成了探针,以不同来源的RNA为模板与制备的探针分别进行了DNA-RNA杂交.S1 mapping的结果表明,来自C85/pIJ4470,C85/pIJ4471,C70/pIJ4470,C70/pIJ4471及C17/pIJ4470,C17/pIJ4471菌株的RNA杂交后都给出了较强的阳性杂交信号,而来自C71/pIJ4470,C71/pIJ4471菌株的RNA杂交未出现阳性信号,来自C119/pIJ4470与C119/pIJ4471菌株的RNA杂交后有弱的信号.上述结果表明启动子P_TH4和P_TH270的转录依赖于链霉菌分化关键基因whiG,部分依赖于分化基因whiH,而不依赖于分化基因whiA,whiB及whiI.     

Identification of the cardiac and circulating form of atriopeptin in rabbit

Analysis of peptides purified from high and low molecular weight fractions of rabbit atrial extracts indicates that the sequence of the first 30 residues of rabbit atriopeptigen exhibits 80% homology with the rat peptide, and that the low molecular weight rabbit peptide (28 residues) is identical to rat atriopeptin 28 (AP 28). The effects of infused 1-deaminoarginine8-vasopressin (dAVP) and phenylephrine, volume expansion, and water immersion on AP release into the circulation of the rabbit was studied. Neither dAVP, nor water immersion elevated right atrial pressure (RAP) or plasma AP levels in the anesthetized rabbits. Phenylephrine induced a sustained increase in systemic blood pressure and right atrial pressure which was accompanied by elevated plasma AP immunoreactivity which appeared to be identical to rat AP-28 on HPLC. There is obviously a preferential conservation of the AP sequence, since the C-terminal peptide is exactly the same in rabbit, rat and mouse and differs from human, dog, cow and pig only by the single substitution of an isoleucine for a methionine residue.     

北京地区红隼的迁徙路线和活动区域

红隼(Falco tinnunculus)被列为国家二级重点保护野生动物,是能同时适应农村和城市环境的小型猛禽,对维持城市生态系统稳定具有重要意义。2022年4月至7月,为在北京救助的7只红隼佩戴了卫星追踪器,追踪其活动轨迹,依据追踪的动物活动位点数据,采用净平方位移-时间曲线依次对各红隼的迁徙模式进行了判别,深入分析了迁徙红隼的迁徙时间、距离和路线等,并采用核心密度法分别计算了迁徙及留居型红隼95%及50%活动区面积。研究结果表明,在北京地区红隼的迁徙模式为部分迁徙,追踪的7只红隼个体(N01 ~ N07)中,4只为留鸟,1只为迁徙鸟,2只居留类型无法准确判断。N01为迁徙红隼,其度夏地和越冬地分别在内蒙古锡林郭勒盟和河北廊坊,此红隼秋季迁徙速度明显高于春季,其春季迁徙距离551 km,历时25 d,平均迁徙速度为22 km/d,而秋季迁徙距离412 km,历时2 d,平均迁徙速度为203 km/d,河北承德滦平县是其春季迁徙的重要中途停歇地。不同红隼个体间95%及50%活动区面积均存在较大差异,迁徙红隼N01 95%、50%活动区面积在度夏区分别为93.10 km²、17.50 km²,在越冬区分别为7.03 km²、0.99 km²;留居型红隼95%、50%活动区面积均值分别为1 165.34 km²、178.71 km²(n = 4),其中最大95%、50%活动区面积分别为4 320.26 km²(N02)、648.22 km²(N02),最小95%、50%活动区面积分别为2.80 km²(N03)、0.29 km²(N03)。本研究揭示了北京地区红隼的迁徙模式、迁徙路线、重要停歇地及活动区状况,为红隼的针对性保护和管理提供了科学依据。     

全球啄木鸟濒危格局及研究现状

啄木鸟科物种作为初级洞巢者与蛀干害虫控制者,对森林高度依赖,是森林生态系统重要的伞护种和环境指示物种。自20世纪以来,由于全球范围的栖息地丧失和片段化,啄木鸟科物种的森林生境急剧萎缩,威胁着该类群物种的生存和繁衍。为探究啄木鸟科动物濒危情况和研究现状,本研究利用世界自然保护联盟濒危物种红色名录(IUCN Red List)和国际鸟盟(BirdLife International)在线数据库,检索并整理出1988年到2023年以下内容：(1)全球啄木鸟的物种数、濒危等级及其变化情况;(2)各大洲的啄木鸟物种数及其受威胁物种的比例;(3)啄木鸟的主要威胁因素;(4)通过Google学术搜索等方式检索并统计啄木鸟相关文章的研究内容。结果显示：(1)目前现存254种啄木鸟,33年间全球受威胁啄木鸟由7种增加至18种,受威胁物种数占当年已命名啄木鸟物种数的比例由3.4%上升至7.0%。(2)亚洲、南美洲和北美洲各分布了83种、93种和56种啄木鸟,受威胁物种占比分别为12.0%、6.4%、5.3%。非洲和欧洲分别分布了36种和11种啄木鸟,当前没有受威胁物种。(3)农业和生物资源利用以及放牧是啄木鸟的主要威胁因素。(4)共检索到研究啄木鸟的有关文章1 024篇,研究覆盖了140种啄木鸟,其中,文章数最多的物种是红顶啄木鸟(Leuconotopicus borealis)(162篇)。研究主要集中在巢相关特征(129篇)、生境选择特征(122篇)、取食行为(112篇)、繁殖行为(99篇)和种群状况(66篇)等基础生态学内容。这些研究为啄木鸟生物学、生态学积累了一定的基础,但物种覆盖程度还远远不够。在生物多样性急剧丧失的大背景下,亟需开展更为广泛和深入的研究。本研究对全球啄木鸟的濒危格局与研究现状进行了全面的分析,以期为后续啄木鸟的研究与保护工作提供参考。     

通过染色体外同源重组建立乳腺中表达外源基因的转基因小鼠

为研究染色体外重组方法在创建转基因动物中的应用,选取牛asl酪蛋白基因的5′及3′侧翼区和氯霉素乙酰化酶编码区,构建了两个具有3 kb相互重叠的融合基因.将这两个DNA片段末端脱磷酸后,以摩尔比1:1的比例混合,通过显微注射导入小鼠受精原核,最后获得了11个品系的转基因小鼠.对其中10个品系的小鼠的整合分析表明,所注射的两个DNA片段均发生了染色体外同源重组,而且除了 1个品系的小鼠丢失了大约1kb的序列外,其余品系小鼠的重组产物与预想的结构符合.在当代和后代的转基因小鼠乳汁中均可测到氯霉素乙酰化酶的活性.这表明融合基因在转基因小鼠乳腺中得到表达和分泌,也说明显微共注射两个相互重叠的基因片段是建立转基因动物的一个可行途径.     

转基因红花中角质细胞生长因子KGF-1的表达

通过构建重组表达质粒载体p139035S-KGF1和根癌农杆菌介导在红花(Carthamus tinctorius)中表达角质细胞生长因子(KGF-1)。从侵染到诱导生根共需要14周, 转化率达0.1%。红花子叶在潮霉素筛选培养基上培养4–5周后便可获得丛生芽, 再生芽移入含潮霉素的伸长生根培养基, 培养4–8周可诱导生根。通过PCR、Southern blot、RT-PCR及Western blot检测证明目的基因KGF-1已经整合到红花细胞的染色体中, 实现了KGF-1外源蛋白在红花中的成功表达, 为开发KGF-1蛋白新的生产途径奠定了基础。     

Chaperone-mediated refolding of recombinant prochymosin

It has been verified that prochymosin is characterized by a two-stage refolding: dilution of unfolded protein into pH 11 buffer followed by neutralization at pH 8; the high-pH step is indispensable. Here we demonstrate that one-stage refolding around pH 8 can be achieved when GroE or 10-fold molar excess (rather than catalytic concentration) of protein disulfide isomerase (PDI) over prochymosin is present. The helping effect varies with the oxidation states of prochymosin. GroE and PDI increase the reactivation of the unfolded, partially reduced and the unfolded, oxidized prochymosin from 5% to 40% and from 50% to 100%, respectively. For the unfolded and fully reduced prochymosin, GroE does not have a positive effect, whereas PDI promotes renaturation from 2% to 28%. Based on our previous and present observations, we propose that at pH 8 there may be two kinds of incorrect interactions within and between prochymosin polypeptides leading to unproductive pathways: one prevents disulfide rearrangement, which can be avoided by high pH; the other interferes with acquisition of native conformation, which can be relieved by GroE and PDI.     

A Passive Anti-icing Strategy Based on a Superhydrophobic Mesh with Extremely Low Ice Adhesion Strength

Although superhydrophobic materials have attracted much research interest in anti-icing,some controversy still exists.In this research,we report a cost-effective method used to verify the contribution of area fraction to ice adhesion strength.We tried to partially-embed siliea nanopnarticles into microscale fabrics of a commercial polyamide mesh.Then,the area fraction could be determined by altering the mesh size.Generally,the ice adhesion strength decreases as the area fraction decreases.An ice adhesion strength of~1.9 kPa and a delayed freezing time of~1048 s can be obtained.We attribute the low ice adhesion strength to the combination of superhydro-phobicity and stress concentration.The superhydrophobicity prohibits the water from penetrating into the voids of the meshes,and the small actual contact area leads to stress concentration which promotes interfacial crack propagation.Moreover,our superhydrophobic mesh simultaneously exhibis a micro-nano hierarchical structure and a partally-cmbedded structure.Therefore,the as-prepared superhydrophobic mesh retained the ieephobicity after 20 icingldeicing cycles,and maintained its superhydrophobicity even afier 60 sandpaper-abrasion cycles and a 220"C thermal treatment.     

Glymphatic distribution of CSF-derived apoE into brain is isoform specific and suppressed during sleep deprivation

Background

Apolipoprotein E (apoE) is a major carrier of cholesterol and essential for synaptic plasticity. In brain, it’s expressed by many cells but highly expressed by the choroid plexus and the predominant apolipoprotein in cerebrospinal fluid (CSF). The role of apoE in the CSF is unclear. Recently, the glymphatic system was described as a clearance system whereby CSF and ISF (interstitial fluid) is exchanged via the peri-arterial space and convective flow of ISF clearance is mediated by aquaporin 4 (AQP4), a water channel. We reasoned that this system also serves to distribute essential molecules in CSF into brain. The aim was to establish whether apoE in CSF, secreted by the choroid plexus, is distributed into brain, and whether this distribution pattern was altered by sleep deprivation.

Methods

We used fluorescently labeled lipidated apoE isoforms, lenti-apoE3 delivered to the choroid plexus, immunohistochemistry to map apoE brain distribution, immunolabeled cells and proteins in brain, Western blot analysis and ELISA to determine apoE levels and radiolabeled molecules to quantify CSF inflow into brain and brain clearance in mice. Data were statistically analyzed using ANOVA or Student’s t- test.

Results

We show that the glymphatic fluid transporting system contributes to the delivery of choroid plexus/CSF-derived human apoE to neurons. CSF-delivered human apoE entered brain via the perivascular space of penetrating arteries and flows radially around arteries, but not veins, in an isoform specific manner (apoE2?>?apoE3?>?apoE4). Flow of apoE around arteries was facilitated by AQP4, a characteristic feature of the glymphatic system. ApoE3, delivered by lentivirus to the choroid plexus and ependymal layer but not to the parenchymal cells, was present in the CSF, penetrating arteries and neurons. The inflow of CSF, which contains apoE, into brain and its clearance from the interstitium were severely suppressed by sleep deprivation compared to the sleep state.

Conclusions

Thus, choroid plexus/CSF provides an additional source of apoE and the glymphatic fluid transporting system delivers it to brain via the periarterial space. By implication, failure in this essential physiological role of the glymphatic fluid flow and ISF clearance may also contribute to apoE isoform-specific disorders in the long term.