硒蛋白R抑制铜离子介导的Aβ_(42)聚集和细胞毒性

阿尔茨海默病(Alzheimer's disease,AD)是一种神经退行性疾病,其主要病理特征包括脑内出现老年斑和神经纤维缠结以及神经元缺失。过渡金属离子如Fe~(2+)、Zn~(2+)、Cu~(2+)等会在AD病人脑中大量蓄集,从而促进,-淀粉样多肽(Aβ)聚集并产生大量活性氧自由基,引起细胞功能改变。硒蛋白R(Sel R)是一种蛋氨酸亚砜还原酶,可将蛋白中的蛋氨酸亚砜还原为蛋氨酸。然而Sel R与金属离子的关系尚无报道。文章对Sel R的突变体Sel R'(Sel R中硒代半胱氨酸Sec95突变为半胱氨酸Cys95)结合Cu~+的性质进行了表征,同时也对Sel R'调节Cu~+/Cu~(2+)诱导的Aβ42聚集和细胞毒性的能力进行了研究。结果表明,Sel R'可与Cu~+特异性螯合剂BCA竞争结合Cu~+,说明Sel R'具有较强的Cu~+结合能力。采用硫磺素荧光法(Th T),发现Cu~+/Cu~(2+)显著性抑制Aβ42的纤维化、诱导其形成非纤维化聚集体,而Sel R'则能够显著抑制Cu~+/Cu~(2+)诱导的Aβ42聚集形态的改变。细胞实验表明:Sel R'能显著降低Cu~+/Cu~(2+)诱导的Aβ42的细胞毒性。上述结果提示:Sel R在脑内除了还原蛋氨酸亚砜,还可通过与铜离子的结合抑制其细胞毒性作用,从而干预AD的发展。     

17β-雌二醇与1-萘酚对雄性罗非鱼（GIFT Oreochromis niloticus）雌激素效应的比较

在实验室条件下,研究了暴露于不同质量浓度1-萘酚（0.064、0.8、2 mg.L-1）与17β-雌二醇（1、10、100μg.L-1）下,对雄性罗非鱼（GIFT Oreochromis niloticus）性腺系数、血清雌二醇质量浓度的影响及血清中卵黄蛋白原的诱导效应,以期对它们的环境雌激素效应有所了解。研究结果显示：暴露于1-萘酚环境下30 d后,雄性罗非鱼的性腺系数有所降低,当质量浓度为2 mg.L-1时,与对照组相比,差异显著（p〈0.05）,雄性罗非鱼血清中雌二醇的量,此时亦达到最大值,为209.3 pg.mL-1,与对照组相比,差异显著（p〈0.05）;但是,血清中卵黄蛋白原并无变化。暴露于17β-雌二醇环境中的雄性罗非鱼,30 d后,其性腺系数有所降低,当质量浓度为10、100μg.L-1时,与对照组相比,差异显著（p〈0.05）;血清中的雌二醇也随着暴露质量浓度的增多,逐渐增多,当质量浓度为100μg.L-1时,差异极显著（p〈0.01）;血清中卵黄蛋白原的质量浓度明显高于对照组,与暴露质量浓度之间呈剂量效应关系。结果显示,17β-雌二醇具有很强的雌激素效应,会使罗非鱼性腺系数下降,诱导血清中雌二醇、卵黄蛋白原的生成;而1-萘酚会使罗非鱼性腺系数下降,诱导血清雌二醇的生成,但是并不诱导卵黄蛋白原的生成,所以推测其雌激素效应可能较弱,需要进一步研究。     

β-甘露聚糖酶基因在枯草芽孢杆菌中的克隆及表达

从Bacillus subtilis JNA 3-10中克隆出β-甘露聚糖酶基因成熟肽链编码序列manA1和含信号肽的β-甘露聚糖酶基因manA2,在B.subtilis 168中克隆表达,分别筛选获得高效分泌表达β-甘露聚糖酶的重组菌株BPM1001(pMA5-manA1/B.subtilis 168)和BPM1002(pMA5-manA2/B.subtilis 168),结果表明菌株BPM1002总酶活力是菌株BPM1001的9.65倍,是原始菌株的13.1倍.在基因manA2下游引入His序列克隆出β-甘露聚糖酶基因manA3,获得枯草芽孢杆菌168重组菌株BPM1003.采用Ni-NTA柱纯化重组菌株BPM1003分泌表达的β-甘露聚糖酶,并研究其酶学性质,该酶促反应的最适pH为6.5,最适温度为65℃,在37℃条件下保存一个月酶活力依然保留有77.8%.5 L发酵罐放大实验结果表明魔芋粉对于产β-甘露聚糖酶具有明显的诱导作用,酶活力最高可达2 748.82 U/mL.图9表3参19     

Biological activity of a red-tide alga-A. tamarense under co-cultured condition with bacteria

The relationship between Alexandrium tamarense （Lebour） Balech, one of red-tide alga, and two strains of marine bacteria, Bacillius megaterium（S7 ） and B. halmapulus（ S10 ） isolated from Xiamen Western Sea, was investigated by evaluating the growth state of A. tamarense and the variation of β-glucosidase activity in co-culture system. The results showed the growth and multiplication of the alga were related with the concentration, genus speciality of the bacteria, and growth stage of the alga itself. The growth of A. tamarense was obviously inhibited by S7 and S10 at high concentration. Either inhibition or promotion contributed much more clearly in earlier than in later stageof the growth of the alga. Furthermore, there was a roughly similar variation trend of the activity of extra-cellular enzyme, β- glucosidase, in the water of the separately co-cultured bacteria S7 and S10 with the alga. The β-glucosidase activity（β-GlcA） rapidly increased during the later algal growth accompanying the increase of the lysis of the alga cells. The obvious inhibition of A. tamarense by marine bacteria at high concentration and evident increase of β-GlcA in co-colture system would help us in better understanding the relationship between red-tide alga and bacteria, and also enlightened us the possible use of bacteria in the bio-control of red-tide.     

Effects of β-cyclodextrins on the enzymatical hydrolysis of chiral dichlorprop methyl ester

The effect of β-cyclodextrins(β-CDs) on the enzymatical hydrolysis of chiral dichlorprop methyl ester (DCPPM) was studied.Four kinds of β-cyclodextrins(β-cyclodextrin, Partly methylated-CD( PM-β-CD), hydroxypropyl-cyclodextrin(HP-β-CD) and carboxymethylcyclodextrin(CM-β-CD) ) were used. Compared with 100% DCPPM in the absence of β-cyclodextrins, the activity of lipase decreased with the increase of β-cyclodextrin and PM-β-cyclodextrin. However, CM-β-cyclodextrin stimulated the lipase activity. The inhibition effect of β-cyclodextrin and PM-β-cyclodextrin on the hydrolysis of DCPPM is affected by many factors other than degree of the methylation blocking the active site of lipase. UV-Vis and Fourier transform infrared(FTIR) spectroscopy studies of the complexation of aqueous DCPPM with β-CDs provide fresh insight into the molecular structure of the complex and explain the effects of β-CDs on enzymatical hydrolysis of chiral DCPPM. Data showed that inclusion complexes had formed by complexation of the CM-β-CD with DCPPM and the solubility of DCPPM was increased in water, which leaded to the increased lipase activity.     

基于ELISA的受体竞争结合法检测环境雌激素(Competitive Estrogen Receptor Binding Based ELISA for Detection of Estrogen in Environment)

为建立环境雌激素的体外快速筛选方法,采用免疫测定介导受体竞争结合试验定量检测环境雌激素.将17β-雌二醇-BSA固定在微孔板上,环境雌激素与微孔板上的17β-雌二醇-BSA竞争性地与雌激素受体结合,与微孔板上17β-雌二醇-BSA结合的受体与雌激素受体的抗体、雌激素受体抗体的二抗形成一个三明治形式的复合物,通过显色剂显色,最后用比色法对试验结果进行定量.结果表明,环境雌激素的浓度与溶液吸光度值成负相关.定量研究可以得到环境雌激素剂量-效应关系曲线,在一定浓度范围内(10ng·L-1～100μg·L-1),溶液的吸光度值与环境雌激素的浓度直接呈现良好的线性关系(R2=0.9583).利用这种方法能检测到的标准品雌二醇的最低浓度为10ng·L-1.以上结果表明,本方法检测环境雌激素操作简便,具有广泛的应用前景.     

Extraction of 17p-estradiol in water using non-imprinted polymer submicron particles in membrane filters

17β-Estradiol (E2) is an endocrine disrupting chemical of harm to both animals and human beings at a low concentration level (ng/L).It cannot be completely removed by wastewater treatments,and is often detected in both environment and drinking waters.The purpose of this feasibility study,towards environmental engineering in the field of water analysis and treatment,was to remove E2 by extraction using non-imprinted polymer (NIP) submicron particles.Experimental results showed that 0.5 mg/L of E2 could be completely extracted by adding 10 mg of NIP particles directly into 10 mL of water.However,the extraction efficiency decreased to 64% for 100 mL of water.prefilling the NIP particles inside a membrane filter showed a potential for water treatment of a large volume,requiring no effort to distribute the particles uniformly in the water.High extraction efficiency (80±10)% for E2 was achieved for 100 mL of water.A total mass of 0.29 mg E2 was extracted from 1000 mL of water containing 0.8 mg/L E2 (by using only 10 mg of NIP particles).Both efficiency and mass capacity can be increased,by scaling up the amount of NIP particles,towards environmental engineering applications.     

钛基β-PbO2阳极催化性能及氧化机理研究

采用提拉法和电沉积法制作Ti／SnO2＋Sb2O3／β-PbO2阳极,通过SEM扫描、XRD分析和极化曲线测定表征具有良好结构和催化性能。利用该电极氧化靛红水溶液,表征Ti／SnO2＋Sb2O3／β-PbO2电极氧化机理,阳极氧化以[·OH]氧化有机物,次级氧化以阳极生成的活性相对差、寿命较长的H2O2,O3氧化有机物。Ti／SnO2＋Sb2O3／β-PbO2阳极氧化有机物是湿式氧化费用的30％～50％,具有广泛应用前景。     

固定化菌剂对污水和牛粪中雌二醇和己烯雌酚的去除作用

针对雌二醇(E2)和己烯雌酚(DES)易在污水和牛粪富集的问题,制备了具有去除E2和DES功能的固定化菌剂,并研究了其对污水和牛粪中E2与DES的去除作用.以E2降解菌Rhodococcus sp.JX-2与DES降解菌Serratia sp.S为供试菌,主要通过正交试验确定了固定化菌剂包埋方法,比较了固定化菌剂、游离态菌剂、空白小球对E2与DES的去除效能,并分析了菌剂用量、p H、含水量、翻堆时间对菌剂去除污水和牛粪中E2和DES的影响.结果表明,菌剂制备的最佳固定化条件为菌株JX-2与菌株S菌悬液体积比为1∶1,海藻酸钠质量分数5%,菌胶比为1∶2,Ca Cl2·2H2O质量分数4%.固定化菌剂对溶液中初始浓度为2 mg·L-1E2和DES的去除率分别达99.42%和84.59%.去除污水中E2和DES的最佳固定化菌剂用量为300g·L-1,p H值为5.0~6.0,能完全去除DES,E2去除率为95.85%;去除牛粪中E2和DES的最佳固定化菌剂用量为600g·kg-1,牛粪含水量为70%,翻堆时间间隔为12 h,能完全去除E2,DES去除率为97.41%.     

Inhibitory kinetics of β-N-acetyl-D-glucosaminidase from Nile tilapia (Oreochromis niloticus) by cadmium北大核心CSCD

Cadmium (Cd2+) pollution in an aquatic environment can negatively affect certain reproductive parameters of aquatic animals. β-N-acetyl-D-glucosaminidase (NAGase) is considered to play an important role in the fertilization process. The aim of the present study was to investigate the effect of Cd2+ on the activity of NAGase purified f rom the testis of Nile tilapia, toward contributing new knowledge on the breadth of negative effects of Cd2+ for Nile tilapia production. The kinetic method of substrate reaction was used for this assessment, and an inhibitory model was established to study the kinetics of NAGase under inhibition by Cd2+. The results showed that Cd2+ could reversibly inhibit the enzymatic activity of NAGase, and the half-maximal inhibitory concentration was estimated to be 40.95 mmol/L. Cd2+ was found to be a competitive inhibitor of NAGase, and the inhibitory constant was determined to be 17.13 mmol/L. The microscopic rate constants of inactivation were also determined. Together, these findings demonstrate that Cd2+ is a reversible inhibitor that can competitively inhibit NAGase. These results may provide a theoretical foundation for further studies on the reproduction of tilapia. © 2018 Science Press. All rights reserved.