Synthesis and bioactive evaluation of novel hybrids of metronidazole and berberine as new type of antimicrobial agents and their transportation behavior by human serum albumin

A series of novel hybrids of metronidazole and berberine as new type of antimicrobial agents were synthesized and characterized by ¹H NMR, ¹³C NMR, IR, MS and HRMS spectra. Bioactive assay manifested that most of the prepared compounds exhibited effective antibacterial and antifungal activities and some showed comparable or superior potency against Methicillin-resistant Staphylococcus aureus to reference drugs Norfloxacin, Chloromycin and Berberine. The transportation behavior of human serum albumin (HSA) to the highly active compound 5g was evaluated and revealed that the association of imidazole derivative 5g with HSA was spontaneous and the electrostatic interactions played important roles in the transportation of HSA to 5g. The calculated parameters indicated that compound 5g could be effectively stored and carried by HSA.     

Exploring the allosteric mechanism of dihydrodipicolinate synthase by reverse engineering of the allosteric inhibitor binding sites and its application for lysine production

Dihydrodipicolinate synthase (DHDPS, EC 4.2.1.52) catalyzes the first committed reaction of l-lysine biosynthesis in bacteria and plants and is allosterically regulated by l-lysine. In previous studies, DHDPSs from different species were proved to have different sensitivity to l-lysine inhibition. In this study, we investigated the key determinants of feedback regulation between two industrially important DHDPSs, the l-lysine-sensitive DHDPS from Escherichia coli and l-lysine-insensitive DHDPS from Corynebacterium glutamicum, by sequence and structure comparisons and site-directed mutation. Feedback inhibition of E. coli DHDPS was successfully alleviated after substitution of the residues around the inhibitor’s binding sites with those of C. glutamicum DHDPS. Interestingly, mutagenesis of the lysine binding sites of C. glutamicum DHDPS according to E. coli DHDPS did not recover the expected feedback inhibition but an activation of DHDPS by l-lysine, probably due to differences in the allosteic signal transduction in the DHDPS of these two organisms. Overexpression of l-lysine-insensitive E. coli DHDPS mutants in E. coli MG1655 resulted in an improvement of l-lysine production yield by 46 %.     

Crystal structure of parallel G-quadruplex formed by the two-repeat ALS- and FTD-related GGGGCC sequence

The hexanucleotide repeat expansion, GGGGCC (G4C2), within the first intron of the C9orf72 gene is known to be the most common genetic cause of both amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). The G4C2 repeat expansions, either DNA or RNA, are able to form G-quadruplexes which induce toxicity leading to ALS/FTD. Herein, we report a novel crystal structure of d(G4C2)₂ that self-associates to form an eight-layer parallel tetrameric G-quadruplex. Two d(G4C2)₂ associate together as a parallel dimeric G-quadruplex which folds into a tetramer via 5′-to-5′ arrangements. Each dimer consists of four G-tetrads connected by two CC propeller loops. Especially, the 3′-end cytosines protrude out and form C·C+•C·C+/ C·C•C·C+ quadruple base pair or C•C·C+ triple base pair stacking on the dimeric block. Our work sheds light on the G-quadruplexes adopted by d(G4C2) and yields the invaluable structural details for the development of small molecules to tackle neurodegenerative diseases, ALS and FTD.     

枸杞体细胞胚发生中蛋白质和DNA代谢动态的立体计量

本文以宁夏构杞无菌苗叶片为材料,离体培养,并诱导体细胞胚发生。根据细胞形态计量学原理,应用数字图像处理软件计量由光学底片经A/D转换成的数字图像中的蛋白质和DNA大分子,对于构杞体细胞胚发生过程中蛋白质和DNA分子的代谢动态进行了量化处理,并对量化结果分析了蛋白质和DNA代谢动态与体细胞胚发生、发育的关系。     

Platelet glycoprotein Ib beta/IX mediates glycoprotein Ib alpha localization to membrane lipid domain critical for von Willebrand factor interaction at high shear

The localization of the platelet glycoprotein GP Ib-IX complex (GP Ibα, GP Ibβ, and GP IX) to membrane lipid domain, also known as glycosphingolipid-enriched membranes (GEMs or raft) lipid domain, is essential for the GP Ib-IX complex mediated platelet adhesion to von Willebrand factor (vWf) and subsequent platelet activation. To date, the mechanism for the complex association with the GEMs remains unclear. Although the palmitate modifications of GP Ibβ and GP IX were thought to be critical for the complex presence in the GEMs, we found that the removal of the putative palmitoylation sites of GP Ibβ and GP IX had no effects on the localization of the GP Ib-IX complex to the GEMs. Instead, the disruption of GP Ibα disulfide linkage with GP Ibβ markedly decreased the amount of the GEM-associated GP Ibα without altering the GEM association of GP Ibβ and GP IX. Furthermore, partial dissociation with the GEMs greatly inhibited GP Ibα interaction with vWf at high shear instead of in static condition or under low shear stress. Thus, for the first time, we demonstrated that GP Ibβ/GP IX mediates the disulfide-linked GP Ibα localization to the GEMs, which is critical for vWf interaction at high shear.     

狂犬病病毒抗体胶体金检测试纸的制备

通过胶体金免疫层析技术建立一种特异、便捷、快速的狂犬病病毒抗体检测方法,对犬等动物免疫狂犬病疫苗后的抗体水平监测提供参考。用醋酸锌沉淀法沉淀狂犬病病毒CVS11,Sepharose 4FF进行层析纯化。用柠檬酸三钠还原法制备的胶体金,标记纯化的狂犬病病毒,喷于试纸的结合释放垫 (金标垫),将SPA (葡萄球菌表面A蛋白) 和纯化的兔抗狂犬病病毒IgG分别喷于试纸的T (检测线) 处和C (对照线) 处,组装试纸条。用制备的试纸条对261份犬血清进行检测,与快速荧光灶抑制试验 (RFFIT) 检测的结果一致;对已知效价的犬狂犬病毒中和抗体 (VNA) 大于0.5 IU,结果为阳性;对狂犬病病毒中和抗体 (VNA) 小于0.5 IU/mL的血清,结果为阴性。制备的狂犬病病毒抗体胶体金检测试纸检测犬血清抗体,具有特异、便捷、快速的特点,能够检测出狂犬病病毒中和抗体大于0.5 IU的血清,适用于临床犬血清抗体水平监测,具有良好的应用前景。     

两种鲤科经济鱼类水声目标强度的实验测定与应用评估

鱼类目标强度(Target Strength,TS)及其与鱼体长度的关系是渔业声学技术研究和应用的核心内容之一。本文以我国广泛分布的鲤科经济鱼类鲤(Cyprinus carpio)和鳙(Aristichthys nobilis)为研究对象,选择湖北省宜昌市隔河岩水库的一处僻静库湾作为实验地点,于2017年5月1—20日采用绳系实验法和Simrad EY60型(频率120 kHz)分裂波束回声探测仪,在环境干扰小的实验网箱里逐一测量单体实验鱼游动状态下的目标强度,由此构建2种鱼的目标强度与个体大小关系式。用于实验的鲤和鳙均为26尾,其全长分别为19.9—29.6 cm和49.2—74.2 cm,体重分别为175—461 g和1200—4700 g,实验所测TS值分别为–59.35—–45.20 dB和–34.22—–17.49 dB。回归得到实验鱼目标强度(TS,dB)与全长(TL,cm)关系式(1)鲤: TS = 29.84×lgTL – 95.23(R^2 = 0.74,P<0.01),(2)鳙: TS = 35.88×lgTL – 90.33 (R^2 = 0.83,P<0.01)。同时选择一个小型封闭水体(盐龙湖)“赶、拦、刺、张”联合渔法捕捞之前的鱼探仪调查数据作为分析实例,通过本文鳙TS-TL关系式估算全湖鱼类资源总量为89360 kg,折合40.1 g/m^2,捕捞产量(73610 kg)与之较为接近,在一定程度上反映了实验目标强度测定及其与体长关系式的合理性,有助于丰富我国内陆水域鱼类资源水声学评估技术的基础参数。     

Daylength helps temperate deciduous trees to leaf‐out at the optimal time

Global warming has led to substantially earlier spring leaf‐out in temperate‐zone deciduous trees. The interactive effects of temperature and daylength underlying this warming response remain unclear. However, they need to be accurately represented by earth system models to improve projections of the carbon and energy balances of temperate forests and the associated feedbacks to the Earth's climate system. We studied the control of leaf‐out by daylength and temperature using data from six tree species across 2,377 European phenological network ( www.pep725.eu ), each with at least 30 years of observations. We found that, in addition to and independent of the known effect of chilling, daylength correlates negatively with the heat requirement for leaf‐out in all studied species. In warm springs when leaf‐out is early, days are short and the heat requirement is higher than in an average spring, which mitigates the warming‐induced advancement of leaf‐out and protects the tree against precocious leaf‐out and the associated risks of late frosts. In contrast, longer‐than‐average daylength (in cold springs when leaf‐out is late) reduces the heat requirement for leaf‐out, ensuring that trees do not leaf‐out too late and miss out on large amounts of solar energy. These results provide the first large‐scale empirical evidence of a widespread daylength effect on the temperature sensitivity of leaf‐out phenology in temperate deciduous trees.     

利用烟道气培养微藻的机制与应用

微藻生物柴油是唯一有潜力代替传统化石燃料解决交通用油问题的可再生生物能源,但其产业化主要受到微藻培养高成本的制约。工业废气(烟道气)不仅含有大量CO2,还含有硫氧化物(SOx)和氮氧化物(NOx)。因此,利用烟道气培养产油微藻既可以降低微藻生物柴油的生产成本,又可以减少温室气体和污染气体的排放。综述了微藻液体悬浮培养系统吸收、转化CO2、SOX和NOx的机理和利用烟道气培养微藻的研究与实践,基于微藻细胞具有高效吸收、转化CO2、SO2和NOx的能力,提出了建立微藻产油、固碳、脱硫、除硝一体化模式来帮助解决当前能源和环境问题的设想。     

Development and characterization of a novel anti-IgE monoclonal antibody

IgE is the central macromolecular mediator responsible for the progression of allergic reactions. Omalizumab (Xolair) is a humanized monoclonal anti-IgE antibody directed at the FcεRI-binding domain of human IgE, which represents a novel therapeutic approach in the management of asthma. In this study, we developed a monoclonal antibody (7A5) against human IgE via hybridoma technique. Our data showed that 7A5 could inhibit free IgE molecules to bind to receptors without affecting IgE already bound to cellular receptors. Importantly, 7A5 was able to inhibit IgE-induced histamine release of basophilic leukemia cells. Next, the phage display peptide library technology was employed to select peptides binding to 7A5 and a striking peptide sequence motif was recovered, which is homologous to the sequence ³⁹¹KQR³⁹³ within the Cε3 domain of IgE-Fc, Our results further indicated that 7A5 specifically bound to the synthesized peptide “³⁸⁸KEEKQRN³⁹⁴” containing the ³⁹¹KQR³⁹³ motif in IgE-Fc. The epitope of 7A5 was found to be spatially close to the FcεRI-binding site, suggesting that 7A5 binding to IgE might block IgE binding to receptors via steric hindrance. The anti-IgE monoclonal antibody 7A5 may have the potential to be developed as a therapeutic agent for the treatment of allergic diseases.