Construction of a large phage-displayed human antibody domain library with a scaffold based on a newly identified highly soluble, stable heavy chain variable domain

Currently, almost all U.S. Food and Drug Administration-approved therapeutic antibodies and the vast majority of those in clinical trials are full-size antibodies mostly in an immunoglobulin G1 format of about 150 kDa in size. Two fundamental problems for such large molecules are their poor penetration into tissues (e.g., solid tumors) and poor or absent binding to regions on the surface of some molecules [e.g., on the human immunodeficiency virus envelope glycoprotein (Env)] that are accessible by molecules of smaller size. We have identified a phage-displayed heavy chain-only antibody by panning of a large (size, ∼ 1.5 × 10¹⁰) human naive Fab (antigen-binding fragment) library against an Env and found that the heavy chain variable domain (V_H) of this antibody, designated as m0, was independently folded, stable, highly soluble, monomeric, and expressed at high levels in bacteria. m0 was used as a scaffold to construct a large (size, ∼ 2.5 × 10¹⁰), highly diversified phage-displayed human V_H library by grafting naturally occurring complementarity-determining regions (CDRs) 2 and 3 of heavy chains from five human antibody Fab libraries and by randomly mutating four putative solvent-accessible residues in CDR1 to A, D, S, or Y. The sequence diversity of all CDRs was determined from 143 randomly selected clones. Most of these V_Hs were with different CDR2 origins (six of seven groups of V_H germlines) or CDR3 lengths (ranging from 7 to 24 residues) and could be purified directly from the soluble fraction of the Escherichia coli periplasm. The quality of the library was also validated by successful selection of high-affinity V_Hs against viral and cancer-related antigens; all selected V_Hs were monomeric, easily expressed, and purified with high solubility and yield. This library could be a valuable source of antibodies targeting size-restricted epitopes and antigens in obstructed locations where efficient penetration could be critical for successful treatment.     

基于NDVI时间序列轨迹的草原露天矿区植被时空动态特征

采矿作用下草原露天矿区植被时空动态特征尚不明确.以胜利露天矿区为例,选取MODIS和Landsat影像,利用ESTARFM构建2001—2015年植被生长期一致的年际Landsat时间序列,以时间分割算法拟合像元NDVI时间序列轨迹,结合轨迹形态特征提取早期扰动型、持续扰动型、扰动稳定型、扰动稳定恢复型和扰动恢复型5种植被动态类型及各类型的时间特征.结果表明:胜利矿区植被动态类型以扰动恢复型为主,占各类型像元总数的55.2%,扰动稳定型和持续下降型次之,分别为25.6%和11.0%,早期扰动型和扰动稳定恢复型较小,分别为3.5%和4.7%.扰动多发于2004—2009年,稳定期多始于2008年,空间上多分布于露天采场和排土场,恢复期多始于2008年和2010年,其空间范围较小且集中于矿井外围和排土场.扰动持续时长以1年为主,稳定期持续时长以7年为主,恢复期持续时长中稳定恢复型为2～5年,扰动恢复型为8年.     

滦河流域河岸带入侵植物分布特征及其与环境的关系

根据滦河流域河岸带56个样点的植被调查,研究入侵植物种类,其组成特征、空间分布,及其与环境因子的关系.结果表明: 滦河流域河岸带共记录入侵植物26种,属12科19属,其中菊科和苋科植物为优势科,占入侵植物总数的50%,一年生植物占总数的73.1%,反枝苋、大狼杷草、小藜是出现频数最高的3种植物;样点的入侵种数、入侵强度与海拔呈显著负相关;人类活动强度对入侵植物分布有显著影响,人口密集、交通便利、人类活动频繁的平原区,水库众多的浅山区,以及中游段旅游业发达的承德市周边山区是入侵植物集中分布的区域;人口分布较稀的高原地区只出现几种宽生态幅的入侵物种.总体上,滦河流域河岸带入侵等级高的植物主要分布在400 m以下的中低海拔地区,除反枝苋外,其他高等级入侵物种尚未扩散进入高海拔地区.     

Molecular imprinting of nitrophenol and hydroxybenzoic acid isomers: effect of molecular structure and acidity on imprinting

Three nitrophenol isomer-imprinted polymers were prepared under the same conditions using 4-vinylpyridine as a functional monomer. Different recognition capacities for template molecules were observed for the three polymers. Another imprinting system with stronger acidity than nitrophenol isomers, 2-hydroxybenzoic acid (salicylic acid) and 4-hydroxybenzoic acid, was imprinted using 4-vinylpyridine or acrylamide as functional monomer respectively. Both 4-hydroxybenzoic acid-imprinted polymers using the two monomers showed recognition ability for the template molecule. However, when acrylamide was chosen as functional monomer, the salicylic acid-imprinted polymer showed very weak recognition for the template molecule, whereas strong recognition ability of the resultant polymer for salicylic acid was observed with 4-vinylpyridine as functional monomer. It seems that the structure and acidity of template molecules is responsible for the difference in recognition, by influencing the formation and strength of interaction between template molecule and functional monomer during the imprinting process. An understanding of the mechanism of molecular imprinting and molecular recognition of MIPs will help to predict the selectivity of MIPs on the basis of template molecule properties.     

泌尿生殖道非淋菌性感染的病原学以及耐药性分析

目的：探究泌尿生殖道非淋茵性感染的病原学以及耐药性,指导临床合理用药。方法：对我院2006年3月．2011年12月确诊的2136例非淋菌性泌尿生殖道感染患者进行标本采集,对病原体及其耐药性进行检测分析。结果：淋茵感染泌尿生殖道病原茵主要为CT及支原体,男性患者CT感染率高于女性患者,女性患者支原体感染率高于男性患者,此外,念珠茵、滴虫及其他难以检出细菌亦属于致病病原茵的一种;在12种抗生素中,多西环素、米诺环素及交沙霉素敏感率较高,均〉80％,罗红霉素、红霉素及环丙沙星敏感度较低,均〈20％。结论：非淋茵感染泌尿生殖道病原菌主要为CT及支原体,男性患者CT感染率高于女性患者,女性患者支原体感染率高于男性患者,病原茵对多西环素、米诺环素及交沙霉素耐药性较低,但不同地域病原茵耐药性亦存在差异,应按照实际情况进行耐药性检测,指导临床合理用药。     

Rapid method for protein quantitation by Bradford assay after elimination of the interference of polysorbate 80

Bradford assay is one of the most common methods for measuring protein concentrations. However, some pharmaceutical excipients, such as detergents, interfere with Bradford assay even at low concentrations. Protein precipitation can be used to overcome sample incompatibility with protein quantitation. But the rate of protein recovery caused by acetone precipitation is only about 70%. In this study, we found that sucrose not only could increase the rate of protein recovery after 1 h acetone precipitation, but also did not interfere with Bradford assay. So we developed a method for rapid protein quantitation in protein drugs even if they contained interfering substances.     

Process of simultaneous hydrogen sulfide removal from biogas and nitrogen removal from swine wastewater

The feasibility of a new flowchart describing simultaneous hydrogen sulfide removal from biogas and nitrogen removal from wastewater was investigated. It took 30 days for the reactor inoculated with aerobic sludge to attain a removal rate of 60% for H₂S and NO_x–N simultaneously. It took 34 and 48 days to attain the same removal rate for the reactor without inoculated sludge and the reactor inoculated with anaerobic sludge respectively. The reactor without inoculated sludge still operated successfully, despite requiring a slightly longer startup time. The packing material was capable of enhancing the removal efficiency of reactors. Based on the concentration of NO_x–N and H₂S in the effluent, the loading rate and the ability of the system to resist shock loading, the performance of the reactor filled with hollow plastic balls was greater than that of the reactor filled with elastic packing and the reactor filled with Pall rings.     

Study of the restitution of action potential duration using the artificial neural network

It is widely accepted that the APD (action potential duration) restitution plays a key role in the initializing and maintaining of the reentry arrhythmias. The Luo-Rudy II models paced with different protocols showed that the current APD had a complex relation with the previous APDs and diastole intervals (DIs). This relation could not be accurately described by a single exponential function. We used an artificial neural network to formularize this relation. The results suggested that back-propagation (BP) network could predict the current APD from the information of the first three previous beats. This would help provide a target for potential anti-arrhythmic therapies.