Characterization of the Antigenicity and Immunogenicity of the Escherichia Coli Produced RNase Domain of the Classical Swine Fever Virus Glycoprotein E~(rns)

Erns is a highly glycosylated envelope protein of classical swine fever virus (CSFV) with RNase activity. Erns can induce neutralizing antibodies and provide immune protection against CSFV infection. In this study, the RNase domain of the Erns was produced in Escherichia coli. Its reactivity with CSFV-positive sera and its ability to induce antibodies and to provide protective immunity were then investigated. The serological tests showed that the prokaryotically expressed RNase domain of the Erns retained i...     

Induction of multi-epitopespecific antibodies against HIV-1 by multi-epitopevaccines

Some neutralizing antibodies against HIV-1 envelope proteins were highly effective to inhibit the infection of different strainsin vitro, and existed in the infected individuals with very low levels. We suggested multi-epitope-vaccine as a new strategy to increase levels of neutralizing antibodies and the abilities against HIV mutationin vivo. Two candidate multi-epitope-vaccines induced antibodies with predefined multi-epitope-specificity in rhesus macaque. These antibodies recognized corresponding neutralizing epitopes on epitope-peptides, gp41 peptides, V3 loop peptide, rsgp41 and rgp120. Besides, three candidate epitope-vaccines in combination (another kind of multi-epitopevaccines) showed similar potency to induce predefined multiple immune responses in rabbits. These results suggest that multi-epitope-vaccines may be a new strategy to induce multi-antiviral activities against HIV-1 infection and mutations.     

The short C-terminal hydrophilic domain of NhaH Na^＋/H^＋ antiporter from Halobacillus dabanensis with roles in resistance to salt and in pH sensing

The Na^＋/H^＋ antiporter plays key roles in maintaining low cytoplasmic NaNa^＋ level and pH homeostasis, while little is known about the Carboxyl-terminal hydrophilic tails of prokaryotic antiporters. In our previous study, the first Na^＋/H^＋ antiporter gene nhaH from moderate halophiles was cloned from Halobacillus dabanensis D-8 by functional complementation. A topological model suggested that only nine amino acid residues （^395PLIKKLGMI403） existed in the hydrophilic C-terminal domain of NhaH. The C-terminal truncated mutant of NhaH was constructed by PCR strategy and designated as nhaH△C. Salt tolerance experiment demonstrated that the deletion of hydrophilic C-terminal nine amino acid residues significantly inhibited the complementation ability of E. coil KNabc, in which three main Na^＋/H^＋ antiporters nhaA, nhaB and chaA were deleted. Everted membrane vesicles prepared from E. coil KNabc/nhaHAC decreased both Na^＋/H^＋ and Li^＋/H^＋ exchange activities of NhaH, and also resulted in an acidic shift of its pH profile for Na^＋, indicating a critical role of the short C-terminal domain of NhaH antiporter in alkali cation binding/translocation and pH sensing.     

A possible configuration of H 5 + cluster

On the basis of our previous work, a configuration of H ₅ ⁺ has been presented that is a square center structure, and the energy for the structure of H ₅ ⁺ has been obtained by IACQM. The result showed that there is an extreme energy E_e: −2.45981 [2 Ry] atR = 2.50a ₀. It indicates that the square center structure of H ₅ ⁺ system has the certain stability, which is a possible configuration of H ₅ ⁺ .     

Spectroscopic investigation of the interaction between human serum albumins and 10-hydroxycamptothecin

The binding reaction between 10-hydroxycamptothecin (10-HCPT) and human serum albumins (HSA) is studied by means of fluorescence spectroscopy, UV-Vis absorption spectrum, 1H NMR spectrum, and molecular simulation. The results indicate that the binding reaction of 10-HCPT and HSA is a single static quenching process, and the binding equilibrium constant for 10-HCPT binding with HSA is estimated K ₀= 4.93×10⁴ L · mol⁻¹ at 25 °C with the molar ratio of 1:1. The distance (r) and energy transfer efficiency (E) between donor (HSA) and acceptor (10-HCPT) are obtained as follows, r =3.51 nm; E =0.27. The enthalpy change (ΔH ^⦵) and entropy change (ΔS ^⦵) are calculated at different temperatures, and the hydrophobic force and shidipole force are the functions in the reaction. The results show that 10-HCPT binds within the subdomain II A of HSA by the hydrophobic force, and the 10-OH and 20-OH of 10-HCPT bind with both residue Leu-238 of HSA and Ala 291 of HSA by hydrogen bonds. Biography: LI Guizhi(1962–), female, Associate professor, research direction: organic analysis.     

Cloning and functional analysis of the sequences flanking mini-Tn5 in the magnetosome-deleted mutant NM21 of Magnetospirillum gryphiswaldense MSR-1

A magnetosome-deleted mutant NM21 of Magnetospirillum gryphiswaldense MSR-1 was generated by mini-Tn5 lacZ2 transposon mutagenesis, and a 3073-bp fragment flanking mini-Tn5 lacZ2 in NM21 was cloned by Anchored PCR. Sequencing analysis showed that this fragment involved three putative ORFs; the mini-Tn5 lacZ2 was inserted into ORF1. Functional complementary test indicated that the 3073-bp fragment was required for biosynthesis of magnetosomes in M. gryphiswaldense MSR-1. The majority of proteins, which had homology with the protein encoded by ORF1, were the cation transporter. Transmembrane domain analysis showed that the protein encoded by ORF1 contained four transmembrane domains. It may be a transmembrane protein. The protein encoded by ORF1 contained two putative conserved domains: COG0053 and PRK09509. The MMT1 and FieF, containing conserved domains COG0053 and PRK09509 too, were Fe²⁺ transporter (cation diffusion facilitator superfamily). It was suggested that the protein encoded by ORF1 might take part in the magnetosomes biosynthesis as Fe²⁺ transporter. Supported by National Natural Science Foundation of China (Grant No. 30570023) and Scientific Research Project of Huaibei City, Anhui Province (Grant No. 070114)     

黑鲷NKCC1分子特征及其对急性盐度胁迫的表达响应

为了解黑鲷(Acanthopagrus schlegelii)在盐度胁迫过程中的适应机制,本研究利用基因扩增、聚类分析、荧光定量PCR等技术对NKCC1基因进行生物信息学分析以及在不同组织中的表达特征研究,并探讨其在急性盐度胁迫下的表达机制.结果表明NKCC1基因的开放阅读框(Open Reading Frame,OR...     

The features of microcontroller EM78P447 and technique of port connection with I2C series bus

By describing the technique of port connection and flow chart ofR/W operation between 24LC01, serial E²PROM, and EM78P447A, features of EM78P447 and its connection with I²C serial bus are introduced. In the end, the programe of writing operation is given. Foundations item: Research on Electromagnetic Scatter Character of very Complicate Object (69571021) Biography: Huang Ji-wu(1939-), male, Associate professor, research direction: control with microcomputer and intelligent system.     

Export of particulate organic carbon estimated from234Th-238U disequilibria and its temporal variation in the South China Sea

Activities of²³⁴Th and nutrient concentrations in the upper 500 m water column were measured at a time-series station in the South China Sea over a time span of 12.3 d. Results showed a reduction of dissolved²³⁴Th and an overall increase of particulate²³⁴Th during the period. Meanwhile, activities of total²³⁴Th kept fairly constant, implying rapid transformation of²³⁴Th between the dissolved and particulate forms. Vertical profiles of total²³⁴Th showed evident deficit of²³⁴Th relative to²³⁸U in the upper 500 m water column. Using an irreversible steady-state model of thorium scavenging, export fluxes of particulate organic carbon (POC) corresponding to time pointsT ₁ andT ₂ were estimated to be 46.5 and 13.1 mmolC · m⁻² · d⁻¹. It was demonstrated that the estimation of POC export was greatly dependent on the POC/²³⁴Th_P ratios and the bias caused by the different models of²³⁴Th scavenging, however, was considered to be of minor importance.     

含Sr2+和Bi3+有机杂化体的合成及其光降解罗丹明B

采用溶剂热法在N,N-二甲基甲酰胺溶液中合成了不同摩尔比例的SrCl_2和Bi(NO_3)_3(分别为5%,10%,30%,50%,80%)与对苯二甲酸形成的含Sr~(2+)和Bi~(3+)有机杂化体,并在紫外可见光照射下对其进行了罗丹明B降解活性的评价.结果表明:杂化体在紫外可见光下对RhB降解活性良好,其中SrCl_2摩尔比例10%掺杂的催化剂光降解活性最佳,这归结于其在紫外区域对光有很好的吸收及光生电子和空穴能有效地分离.双金属离子能协同调节有机杂化体的光催化活性.     

Construction and expression of retroviruses encoding dual drug resistance genes in human umbilical cord blood CD34+ cells

A series of retro viral vectors encoding humanmdr1 gene alone as wetl as in combination with either humanmgmt gene or human mutantSer ³¹-dhfr gene are engineered. The resultant retroviruses are used to transduce human umbilical cord blood CD34⁺ cetls. It has been shown that expression of dual drug resistance genes in transduced cetls confers a broad range of resistance to both kinds of corresponding drugs. These data suggest a rationale for the use of such double chemoresistance gene constructs in anin vivo model in which transduced hematopoietic cetls will acquire multiple protection against the cytotoxic side effects of combination chemotherapy and may have future application in chemoprotection of normal tissues, thus killing tumor cetls more effectivety.     

Expression, distribution and function of the focal adhesion kinase (pp125FAK) during murine ectoplacental cone outgrowthin vitro

Mouse embryo implantation is a complex process that includes trophoblast cells derived from ectoplacental cone (EPC) adhesion to and migration through the extracellular matrix (ECM) of uterine endometrium and invasion into the decidua. At the time of implantation, fibronectin (FN) is abundant in the decidua and is distributed pericellularly around each individual stromal cell, and its receptor (integrin α-5β-1) expression on trophoblast populations is up-regulated. The focal adhesion kinase, a 125 ku protein tyrosine kinase (pp125 FAK), is tyrosine phosphorylated upon integrin engagement with its ECM ligand, and its tyrosine phosphorylation sites then serve as the binding sites which couple it with cellular proteins that contain Src SH2 or SH3 domains. Through these linkages, pp125 FAK may integrate multiple signals triggered by integrins. The model of EPC culture %in vitro% was used to study the expression, distribution and function of pp125 FAK during EPC outgrowth on FN. Results indicated that, pp125 FAK primarily expressed and distributed in cellular focal adhesions of the front edge of trophoblast outgrowth from EPC, and was localized in the peripheral region of the individual migrating trophblast cell; antibody or antisense oligodeoxynucleotide to pp125 FAK inhibited EPC attachment and outgrowth, as well as trophoblast cells spreading and migration. This experiment demonstrated that pp125 FAK as an integrin-mediated signaling molecule was involved in EPC outgrowth %in vitro%, and played an important role during trophoblast cells interaction with FN.     

广西乐业天坑群中苔藓的放射性核素分析

为探究广西喀斯特地貌对放射性核素分布及当地环境放射性的影响,利用高纯锗探测器对广西喀斯特地区的大曹天坑和邓家坨天坑采集来的苔藓样品中⁷Be、⁴⁰K、²³⁸U、²³²Th、²²⁶Ra、²¹⁰Pb和¹³⁷Cs的放射性比活度进行测量与分析。结果表明：在苔藓植物中, ⁷Be的放射性比活度为202.3-698.8 Bq/kg [平均值：(430.7±169.7) Bq/kg,n=7];⁴⁰K为33.7-159.9 Bq/kg [平均值：(91.6±44.1) Bq/kg, n=7];²³⁸U为检测限(3.0 Bq/kg)至54.4 Bq/kg [平均值：(16.9±23.6) Bq/kg,n=7];²³²Th为1.2-42.5 Bq/kg [平均值：(14.7±15.9) Bq/kg, n=7];²²⁶Ra为0.7-48.4 Bq/kg [平均值：(16.9±20.3) Bq/kg,n=7];²¹⁰Pb为284.1-950.5 Bq/kg [平均值：(555.5±231.0) Bq/kg, n=7]; ¹³⁷Cs为检测限(0.1 Bq/kg)至2.3 Bq/kg [平均值：(1.1±0.9) Bq/kg,n=7]。对比后发现所有测得的数据均在正常范围内,表明天坑这一喀斯特地貌对放射性核素的分布并没有显著影响,该地区也不存在其他污染来源,同时为我国南方喀斯特地区大气示踪提供数据支撑。     

乌天麻外观性状与特征性成分含量相关性研究

为探究乌天麻(Gastrodia elata f.glauca)外观性状与特征性成分含量之间的关系,本研究通过采集贵州、陕西两地天麻主产区的乌天麻鲜品,测定其长度、宽度、厚度、质量等指标,利用电子感官仪测定样品色、气、味等性状特征值,采用高效液相色谱法测定样品中主要有效成分的含量,以进一步探究两者的相关性。结果表明,乌天麻形、色、气、味等性状特征值与其主要成分含量之间均存在一定的相关性。其中,厚度与天麻素、对羟基苯甲醇、巴利森苷类含量的关联性最强,呈正相关;颜色值a~*与天麻素含量呈极显著负相关,L~*、E~*_ab与对羟基苯甲醇含量呈显著负相关;对羟基苯甲醇含量与电子舌CTS (咸)、ANS (甜)响应值呈极显著正相关,与电子鼻LY型响应值呈显著或极显著负相关,与T型和P型响应值呈显著或极显著正相关。研究结果可为乌天麻质量评价提供参考。     

Site-directed mutagenesis reveals new and essential elements for iron-coordination of the sulfur oxygenase reductase from the acidothermophilic Acidianus tengchongensis

Previous study on refolding of sulfur oxygenase reductase (SOR) inclusion bodies from recombinant Escherichia coli showed that iron was critical to the activity of the SOR from Acidianus ambivalens. In this study, enzymatic assays showed that 2,2′-Dipyridyl, Tiron and 8-hydroxyquinoline, which are specific for chelating ferrous or ferric ions, strongly inhibited the activity of SOR from A. tengchongensis, suggesting that iron atom is essential for SOR activity. Alignment of several functionally identified SORs and SOR-like sequences from genome database revealed a conserved, putative iron binding motif, H⁸⁶-X₃-H⁹⁰-X _n -E¹¹⁴-X _n -E¹²⁹ (numbering according to the Acidianus tengchongensis SOR sequence). Three mutants of SOR were generated by site-directed mutagenesis of H₈₆, H₉₀ and E₁₂₉ into phenylalanine or alanine residue in this study. Circular dichroism spectrum determination indicated that there was no change of the secondary structures of mutant SORs, H⁸⁶F, H⁹⁰F and E¹²⁹A, but all mutants were completely inactive. Through determination of iron contents we found that SOR mutants of H⁸⁶F, H⁹⁰F and E¹²⁹A completely or partially lost iron, while mutants of C³¹S, C¹⁰¹S, and C¹⁰⁴S (generated in a previous study) did not. This result indicated that H⁸⁶, H⁹⁰ and E¹²⁹ but not C³¹, C¹⁰¹, and C¹⁰⁴ were involved in binding to iron atom. Based on this and previous studies, it is proposed that the conserved motifs, C³¹-X _n -C¹⁰¹-X₂-C¹⁰⁴ and H⁸⁶-X₃-H⁹⁰-X₂₃-E¹¹⁴-X₁₄-(E/D)¹²⁹, are respectively for sulfur and molecular oxygen binding and activation. These two conserved motifs are essential elements for the SOR activity. Supported by National Natural Science Foundation of China (Grant Nos. 30670018, 30621005) and State Key Basic Research and Development Program of China (Grant No. 2004CB719602)     

广西珍珠湾红树林区中华乌塘鳢的时空分布特征

为了解广西珍珠湾红树林区中华乌塘鳢的天然资源量及其时空变化动态,为红树林生态系统生物多样性的保护和修复提供基础数据,本研究在广西珍珠湾红树林区的内、中、外滩各设置一个圆形封闭定置围网,于2013年的4季度（1月、4月、7月、10月）各选择3 d投放被标志的中华乌塘鳢（Bostrychus sinensis,区别于野生鱼）至围网中,并在次日用诱捕网捕获,基于标志重捕法的原理评估野生中华乌塘鳢的天然资源量。结果显示,珍珠湾红树林区中华乌塘鳢的全年平均种群密度为（18.74±4.04） ind./hm²,平均生物量为（1.44±0.37） kg/hm²,其中1月的生物量显著高于10月,其余时间的种群密度和生物量均无显著差异;其种群密度和生物量在红树林中无显著空间差异。红树林为中华乌塘鳢提供繁育场,生境的破坏和过度捕捞导致中华乌塘鳢天然资源量下降,应加强监管、保护和相关研究,促进中华乌塘鳢资源的恢复。     

~(99)Tc靶件中子学特性及释热率特点的初步研究

99Tc是一种重要的长寿命裂变产物,半衰期长,极易随地下水迁徙扩散.将99Tc嬗变为短寿命或稳定核素,可有效降低其对环境的危害.99Tc在热中子谱下具有较大的吸收截面,采用PWR嬗变99Tc是一种可行途径.为了评估99Tc嬗变靶件对PWR堆芯中子学参数的影响,并为含嬗变靶件的PWR组件热工水力计算提供数据,本文对含99Tc的17×17 PWR组件进行模拟计算,给出了99Tc靶件加入前后,燃料温度系数、慢化剂温度系数的变化规律;计算了稳态工况下99Tc靶件的轴向释热率.相关结论可为PWR嬗变99Tc研究提供参考.     

Reconstructing sea surface temperature,sea surface salinity and partial pressure of carbon dioxide in atmosphere in the Okinawa Trough during the Holocene and their paleoclimatic implications

The sediment core DGKS9603 collected from the Okinawa Trough was used as research target. By use of unsaturated indexU ^k₃₇ of long-chain alkenone, δ¹³C of POC and of planktonic foraminifera (G. sacculifer), the evolutions of sea surface temperature and partial pressure of carbon dioxide in the atmosphere during the Holocene were reconstructed in the Okinawa Trough. And in combination of δ¹⁸O of planktonic foraminifera, the relative difference of sea surface salinity during the Holocene was also reconstructed. Consequently, three cooling events (E₁-E₃) were identified, each of which occurred at 1.7-1.6, 5.1-4.8 and 8.1-7.4 kaBP (cal), respectively. Of the three events, E₂ and E₃ are globally comparable, their occurrence mechanism would be that the main stream of the Kuroshio Current shifted eastward due to the enhanced circulation of the northeastern Pacific Ocean, which was driven in turn by amplified intensity of sunshine and subsequent enhancement of subtropical high pressure; E₁ corresponds to the Small Ice-Age Event occurring between 1550 and 1850AD in China. In the Oki-nawa Trough, E₁ might be also related to the eastward shift of main stream of the Kuroshio current driven by powerful Asia winter monsoon.

     

Regulation of the inward K+-channels in stomatal guard cells by cytoskeletal microtubules

Patch clamp techniques were applied to investigating the regulation of the inward K⁺-channels inVicia stomatal guard cells by cytoskeletal microtubules. The intracellular addition of either microtubule-disassembling reagent amprophos-methyl (APM) or microtubule-stabilizing reagent taxol resulted in significant inhibition of the inward K⁺-currents across the plasma membranes ofVicia stomatal guard cells. The results suggest that the activation of the inward K⁺-channels in stomatal guard cells requires proper dynamic organization of cytoskeletal microtubules. The regulation of the inward K⁺-channels in guard cells by microtubules may mediate the regulation of stomatal movements by cytoskeleton.