Recent advances in zwitterionic nanoscale drug delivery systems to overcome biological barriers

Nanoscale drug delivery systems(nDDS)have been employed widely in enhancing the therapeutic efficacy of drugs against diseases with reduced side effects.Although several nDDS have been successfully approved for clinical use up to now,biological barriers between the administration site and the target site hinder the wider clinical adoption of nDDS in disease treatment.Polyethylene glycol(PEG)-modification(or PEGylation)has been regarded as the gold standard for stabilising nDDS in complex biological environment.However,the accelerated blood clearance(ABC)of PEGylated nDDS after repeated injections becomes great challenges for their clinical applications.Zwitterionic polymer,a novel family of antifouling materials,have evolved as an alternative to PEG due to their super-hydrophilicity and biocompatibility.Zwitterionic nDDS could avoid the generation of ABC phenomenon and exhibit longer blood circulation time than the PEGylated analogues.More impressively,zwitterionic nDDS have recently been shown to overcome multiple biological barriers such as nonspecific organ distribution,pressure gradients,impermeable cell membranes and lysosomal degradation without the need of any complex chemical modifications.The realization of overcoming multiple biological barriers by zwitterionic nDDS may simplify the current overly complex design of nDDS,which could facilitate their better clinical translation.Herein,we summarise the recent progress of zwitterionic nDDS at overcoming various biological barriers and analyse their underlyingmechanisms.Finally,prospects and challenges are introduced to guide the rational design of zwitterionic nDDS for disease treatment.

Influence of nanosized magnesia on the hydration of borehole-sealing cements prepared using different methods

Gas drainage is an efective technology for gas control in coal mines.A high borehole-sealing quality is the fundamental precondition for efcient gas drainage.The expansibilities of cement pastes used in borehole-sealing processes are critical for the borehole-sealing efect.Nanosized magnesia expansive agents are used to improve the expansibilities of cement pastes and improve the borehole-sealing efect.Nuclear magnetic resonance spectrometry and scanning electron microscopy were adopted to study the efects of nanosized magnesia on the hydration of borehole-sealing cements used with diferent preparation methods.The results showed that an increase in the mass fraction of the nanosized magnesia promoted cement hydration,and the mass fraction was positively correlated with the promotion efect.The use of diferent preparation methods did not change the water-phase distribution in the cement.When using the wet-mixing preparation method,nanosized magnesia promoted the induction,acceleration,and deceleration periods of hydration;when using the dry-mixing preparation method,the nanosized magnesia promoted the induction period of cement hydration,and the promotion efect was less obvious than that seen when using the wet-mixing method.When using the wet-mixing preparation method,the nanosized magnesia was uniformly dispersed,thus enlarging the surface area of the reaction,which provided more nucleation sites for the hydration products of the cement and therefore accelerated the hydration reaction.When using the dry-mixing preparation method,the nanosized magnesia powders were dispersed nonuniformly and aggregated.Under these conditions,only a few nanosized magnesia particles on the surfaces of the aggregated clusters took part in hydration,so only a small number of nucleation sites were provided for the hydration products of cement.This led to inconsistent hydration of cement pastes prepared using the dry-mixing method.The surface porosity of the cement prepared with the wet-mixing preparation method frst decreased and then increased with increases in the mass fraction of the nanosized magnesia.The cement surface exhibited compact hydration products and few pores,and the surface was relatively smooth.In comparison,the surface porosity of the cement prepared using the dry-mixing method fuctuated with increasing mass fraction of the nanosized magnesia,resulting in a rough cement surface and microfractures on some surfaces.The two preparation methods both reduced the surface porosity of the cement.The wet-mixing preparation was more efective and consistent in improving the compactness of the cement than the dry-mixing preparation.These results provide important guidance on the addition of nanosized magnesia in borehole-sealing engineering and the selection of cement preparation methods,and they also lay a solid foundation for realizing safe and efcient gas drainage.

Room-temperature degradation of o-xylene in simulated air using an online-regenerable plasma-catalysis reactor with low amounts of nanosized noble metals on Co_(3)O_(4)

The plasma catalytic degradation of o-xylene in simulated air was improved by loading low amounts of Pt,Pd,or Au onto Co_(3)O_(4).At room temperature,o-xylene conversion and CO_(x)selectivity using a0.1 wt%Pt/Co_(3)O_(4)catalyst reached 98.9%and 80%,and the energy efficiency was at the top level in comparison with values in the literature.A stable o-xylene degradation performance could be obtained by online regenerating the heat-insulated reactor with a high energy density.After characterization,it was found that the loading of nanosized Pt not only increased the Co^(3+)/Co^(2+)ratio,where the Co^(3+)benefitted the formation of reactive oxygen species,but also conduced Pt^(0)to oxygen activation,resulting in effective promotion of complete o-xylene oxidation.Operando plasma diffuse reflectance infrared Fourier transform spectroscopy demonstrated the complete o-xylene oxidation and proved that Pt played a key role in the complete oxidation of o-xylene.

The Preparation of Nanosized Pd/ZSM-23 Bifunctional Catalysts for n-Hexadecane Hydroisomerization by Employing PHMB as the Growth Modifi er

The development of highly effective metal-zeolite bifunctional catalysts for the hydroisomerization of n-alkanes is a paramount strategy to produce second-generation biofuels with high quality.In this study,polyhexamethylene biguanide hydrochloride(PHMB)is precisely added to the initial gel to synthesize nanosized ZSM-23 zeolites(Z23-x PH).Due to orientation adsorption and steric hindrance effects of PHMB,each sample of Z23-x PH demonstrates enhanced mesoporosity in comparison with the conventional Z23-C zeolite.Furthermore,the Bronsted acid density of the Z23-x PH samples is also signifi cantly reduced due to a reduction in the distribution of framework Al at T2-T5 sites.The corresponding Pd/23-C and Pd/Z23-x PH bifunctional catalysts with 0.5 wt%Pd loading for n-hexadecane hydroisomerization are prepared by incorporating ZSM-23 zeolites as acid supports.According to the catalytic test results,the suitable addition of PHMB can effectively promote the iso-hexadecane yield.The Pd/Z23-2PH catalyst with an n_(PHMB)/n(_Si)molar ratio of 0.002 demonstrates the highest maximum iso-hexadecane yield of 74.1%at an n-hexadecane conversion of 88.3%.Therefore,the employment of PHMB has provided a simple route for the development of highly effective Pd/ZSM-23 catalysts for n-alkane hydroisomerization.

Nanos3与生殖细胞发育分化

小鼠Nanos3基因是果蝇Nanos的同源基因,是Nanos基因家族的一员.Nanos3是一种RNA结合蛋白,靠近其C端有两个非常保守且连续的Cys-Cys-His-Cys特异锌指结构域.研究显示,Nanos3在小鼠生殖细胞中特异表达,不仅在原始生殖细胞(primordial germ cells,PGCs)的维持方面发挥重要作用,而且是一种启动雄性生殖细胞分化程序的内源性因子,其在精子发生中的重要作用已引起越来越多的关注.探讨Nanos3的生物学功能,有助于了解生殖细胞发育过程中的部分重要机制.本文就Nanos3维持生殖细胞更新和原始生殖细胞的维持等作用作一综述.

河南华溪蟹nanos1、nanos2、piwi基因部分序列的克隆及表达特征

为获取河南华溪蟹(Sinopotamon henanense)nanos1、nanos2和piwi基因的表达特征,在前期河南华溪蟹卵巢组织转录组测序的基础上,对生殖相关基因nanos1、nanos2和piwi进行了克隆和分析,首次获得了河南华溪蟹nanos1、nanos2和piwi基因的部分cDNA序列。利用半定量PCR获取了河南华溪蟹nanos1、nanos2和piwi基因的组织表达谱特征和在卵巢发育过程中的表达变化规律。研究结果显示,nanos1基因在卵巢、精巢、鳃、心和肌肉中均有表达;nanos2基因在卵巢、精巢、副性腺、心脏、肠和肌肉中有较高表达,且卵巢中表达量高于精巢中表达量;piwi基因特异表达于卵巢和精巢。卵巢发育中3个基因的表达时序研究显示,nanos1基因从卵巢发育的增殖期、小生长期、大生长期到成熟期表达量逐渐升高,与之相反,nanos2和piwi基因的表达呈现相反的趋势。这些研究结果将为河南华溪蟹nanos1、nanos2和piwi基因的功能分析奠定基础。

日本血吸虫新基因Sjnanos的克隆、表达及免疫保护效果评估

目的扩增、表达日本血吸虫Sjnanos编码基因并评估其重组蛋白诱导的免疫保护效果。方法从42d龄的日本血吸虫成虫中扩增了一个日本血吸虫性别差异表达的基因,GenBank登录号为AY814948,并对其进行生物信息学分析。将该基因亚克隆入原核表达载体pET28a,转化至大肠埃希菌(E.coli)BL21,用异丙基-β-D硫代半乳糖苷(IPTG)进行诱导表达,用纯化的重组蛋白免疫小鼠,ELISA检测其血清特异性抗体效价,蛋白印迹分析检测其抗原性,以看家基因NADH为内参,应用荧光定量PCR技术分析该基因在血吸虫各个阶段的表达状况。结果同源性分析表明,该基因为日本血吸虫的一个新基因,其编码序列的开放阅读框(ORF)为525bp,编码174个氨基酸,理论相对分子量为19.9,PI为8.2。荧光定量PCR技术分析显示该基因在7d、13d、18d、23d、32d、42d虫体均有表达,但在18d和13d虫体的表达量明显高于其他天数的虫体,在雄虫的表达量高于雌虫。成功构建了该基因的重组表达质粒pET28a(+)-Sjnanos,并在大肠埃希菌中获得表达,Western-blot分析表明重组蛋白具有较好的免疫原性,动物免疫保护试验获得了31.4%的减虫率和53.8%的肝脏减卵率。结论获得日本血吸虫新的抗原基因Sjnanos,该基因的重组蛋白在小鼠中诱导了部分免疫保护作用。

nanos基因的研究进展

nanos基因在时空上的表达调控对于个体发育起着重要作用。母源nanos mRNA在卵子中的定位及翻译调控受到严格的控制。果蝇Nanos通过抑制后极hunchback mRNA的翻译建立胚胎后极信号中心,从而建立前后体轴;同时它对果蝇原始生殖细胞的决定及迁移有重要作用;Nanos对于胚后发育中生殖细胞的维持也是必须的。现对nanos基因的mRNA的定位、翻译调控及Nanos结构和功能的研究进展进行了综述。

汽车传动部件用40Cr钢表面Ni-nanoSiC复合镀层的制备

采用电沉积方法在汽车传动部件用40Cr钢表面制备Ni-nanoSiC复合镀层。以复合镀层中SiC质量分数和复合镀层的硬度作为指标,通过正交实验优化施镀工艺参数,得到最佳施镀工艺参数:搅拌速度为300 r/min、镀液中SiC颗粒质量浓度为20 g/L、温度为50℃、阴极电流密度为14 A/dm2。结果表明:采用最佳施镀工艺参数制备的Ni-SiC复合镀层表面平整、组织致密,其磨损机制为轻度磨粒磨损,平均摩擦因数约为0.45,低于40Cr钢的0.6;磨损量约为6.27 mg,相比40Cr钢约降低25.6%。Ni-nanoSiC复合镀层能够提供有效的防护,改善和提高40Cr钢的抗磨损性能。

细粒棘球绦虫nanos基因的分子克隆及序列分析

为了研究细粒棘球绦虫nanos基因的序列及结构并为疫苗及新的药物研制提供靶标,试验采用注释并克隆细粒棘球绦虫nanos基因的方法,并对其序列结构进行分析;利用在线生物信息学软件分析细粒棘球绦虫nanos基因的序列结构及与其他虫属的进化关系。结果表明:细粒棘球绦虫nanos基因ORF为687 bp,编码228个氨基酸,理论分子质量为58 090.6 u,等电位点为5.08;细粒棘球绦虫NANOS蛋白由3个α螺旋和4个β片层组成;发现共有1个丝氨酸位点,1个苏氨酸位点,2个酪氨酸位点,可能的磷酸化位点是7,8,13,18,19,23位。说明该基因可能在细粒棘球绦虫生殖细胞发育过程中发挥着重要作用。

Hydrothermal synthesis of nanosized ZSM-22 and their use in the catalytic conversion of methanol

ZSM‐22 zeolite with different crystal lengths was prepared using a modified hydrothermal method. Rotation speed, Si/Al molar ratio and co‐solvent have important effects on the crystal size of ZSM‐22. The nanosized zeolite samples were characterized by X‐ray diffraction, X‐ray fluorescence, nitrogen adsorption, scanning electron microscopy, temperature‐programmed desorption of am‐monia and solid state nuclear magnetic resonance. The catalytic performance of nanosized ZSM‐22 was tested using the conversion of methanol. Compared to conventional ZSM‐22, the nanosized ZSM‐22 zeolite exhibited superior selectivity to ethylene and aromatics and lower selectivity to propylene. Stability against deactivation was clearly shown by the nanosized ZSM‐22 zeolite. A higher external surface area and smaller particle size make this nanosized ZSM‐22 zeolite attractive for catalytic applications.

团头鲂nanos3基因的克隆鉴定

为了标记团头鲂(Megalobrama amblycephala)的原始生殖细胞(Primordial Germ Cells, PGCs),首次克隆并鉴定了团头鲂nanos3基因(mananos3)。mananos3全长1027 bp,包括48 bp 5′UTR (5′untranslated Region),490 bp 3′UTR和489 bp开放阅读框(Open Reading Frame, ORF)。该基因编码162个氨基酸。通过序列比对发现Mananos3蛋白和其他物种Nanos蛋白一样,存在一个保守的RNA结合功能域,该功能域包含一个锌指基序(Motif)。系统发育树结果显示, Mananos3与鲤(Cyprinus carpio)的Nanos3最为相近。半定量和定量PCR结果表明, mananos3具有较高的母源表达,并在胚胎发育早期高量表达,而在1000细胞期之后表达量逐渐降低。在成体组织中, mananos3仅在卵巢中检测到表达。mananos3和斑马鱼(Danio rerio) nanos3 (zfnanos3)的3′UTR均可以介导绿色荧光蛋白特异标记团头鲂和斑马鱼胚胎发育早期的PGCs,但是mananos3的3′UTR能够更特异地标记团头鲂的PGCs。通过比对mananos3和zfnanos3的3′UTR发现, mananos3的3′UTR中有一个非经典的miR430识别位点(GCACTA)。通过对该位点的突变研究证实其有利于nanos3在非PGCs组织中的降解。综上所述,团头鲂mananos3的3′UTR序列中的非经典miR430识别位点(GCACTA)可能与介导报告基因在PGCs中特异表达相关。

施氏鲟Asnanos1基因序列分析及其在雌雄不同组织中表达

本研究首先从施氏鲟(Acipenser schrenckii)性腺转录组中获得nanos1(Asnanos1)基因全长c DNA,并对其序列的准确性和特征分别进行PCR验证和生物信息学分析。进一步利用荧光定量RT-PCR技术检测Asnanos1基因在雌、雄施氏鲟的性腺、心脏、鳃、脑、肾、肝脏、脾脏等组织中的表达量。利用荧光定量RT-PCR技术检测Asnanos1基因在雌、雄施氏鲟的性腺、心脏、鳃、脑、肾、肝脏、脾脏等组织中的表达量。结果显示:Asnanos1的c DNA全长为1 389 bp,其中,5′非编码区(UTR)为414 bp,3′UTR为279 bp,ORF为696 bp。该基因共编码231个氨基酸。Asnanos1基因在施氏鲟除心脏以外的各组织中均有表达,在雌、雄鱼的鳃、脑、肾、肝脏、脾脏等组织中表达量无显著差异,但在卵巢中的表达量显著高于其他各组织(P<0.05)。所得到的结果可为人工培育全雌施氏鲟苗种提供一些基础资料。

多倍体银鲫nanos2等位多态性、共线性和表达模式分析

为研究生殖干细胞(Germline stem cells, GSCs)的标记基因nanos2的功能,在银鲫(Carassius gibelio)中克隆了两个nanos2的同源基因(Homologs),将其命名为Cgnanos2a和Cgnanos2b,等位、系统进化树和共线性分析表明,在银鲫进化历程中发生了额外的两轮多倍化事件,是一个异源六倍体。qPCR分析表明Cgnanos2a和Cgnanos2b在5月龄银鲫精巢中的表达水平最高,其次是卵巢;对在孵化后25—190d的卵巢中的表达动态分析表明,孵化后25d的卵巢中的Cgnanos2a和Cgnanos2b转录水平最高,随后表达水平急剧下降;并且Cgnanos2a和Cgnanos2b呈现出偏向表达的特征。切片RNA原位杂交实验结果表明, Cgnanos2a和Cgnanos2b均特异地在银鲫卵巢邻近生殖上皮的胞囊(Cyst)中一类直径小于20μm的细胞中表达, Cgnanos2a在精巢精小囊边缘一类单个或两个紧紧相邻的细胞中高表达,推测是银鲫的GSCs。此外,还可在精原细胞和初级精母细胞中检测到Cgnanos2a和Cgnanos2b的转录本。研究通过对nanos2阳性细胞的追踪描绘了银鲫卵巢早期胞囊的发育过程,为后续分离银鲫GSCs奠定了基础;同时对银鲫nanos2两个歧化的部分同源基因的序列特征、进化及表达特征分析,为研究鱼类多次多倍化事件后重复基因的进化提供了一个典型例子。

罗氏沼虾nanos1基因克隆、表达及亚细胞定位

为探究nanos1基因在罗氏沼虾(Macrobrachium rosenbergii)生殖发育中的功能,克隆了罗氏沼虾nanos1基因,其cDNA序列全长2811 bp,编码243个氨基酸;系统进化分析结果表明其与中华绒螯蟹(Eriocheir sinensis)的同源性最高,与鱼类的nanos1同源性较高,与哺乳类同源性较低。半定量PCR检测发现,nanos1在卵巢中特异表达。通过实时荧光定量PCR检测该基因在不同胚胎发育时期及幼体中的表达水平,结果显示:nanso1 mRNA在(卵)未受精时期表达量最高,显著高于(卵)受精期及胚胎发育各期(P<0.05);在胚胎发育期间,(卵)受精时期表达量最高,显著高于卵裂期且极显著高于胚胎发育后期(P<0.01);该基因在卵裂期的表达水平显著高于囊胚期至幼体期;而囊胚期至幼体期之间表达水平较低且无差异。原位杂交技术检测显示,nanos1 mRNA在卵原细胞及初级卵母细胞(Oc1,Oc2,Oc3,Oc4)的细胞质中表达。以上结果表明,nanos1与罗氏沼虾雌性生殖细胞发育有着密切的关系。

黄鳝nanos3基因克隆鉴定及其组织表达分析

【目的】明确nanos3基因在黄鳝卵巢发育和维持过程中的重要作用,为黄鳝PGCs可视化分子标记、生殖细胞移植及性腺发育分化机制研究打下基础。【方法】通过RACE克隆黄鳝nanos3基因cDNA序列,运用ProtParam、SOMPA、SWISS-MODEL、SignalP、TMHMM及NetPhos等在线软件进行Nanos3蛋白生物信息学分析;采用实时荧光定量PCR检测nanos3基因在黄鳝不同组织中的表达情况,并以冰冻切片荧光原位杂交进行性腺组织定位。【结果】黄鳝nanos3基因cDNA序列全长1289 bp,包括147 bp的5’非编码区(5’-UTR)、531 bp的开放阅读框(ORF)及611 bp的3’非编码区(3’-UTR),共编码176个氨基酸残基。黄鳝Nanos3蛋白分子量为19.61 kD,理论等电点(pI)为9.07,为碱性蛋白;在黄鳝Nanos3氨基酸序列第108~162位处存在2个锌指基序“CCHC”结构域,无信号肽和跨膜结构域;蛋白二级结构以无规则卷曲的占比最高(64.20%),其次是α-螺旋(占24.43%),延伸链、β-转角的占比分别为6.82%和4.55%。黄鳝与大刺鳅、射水鱼、斑马鱼等鱼类的Nanos3氨基酸序列相似性分别为67.5%、57.2%和50.3%,与中国林蛙、粗皮蛙等两栖类动物的相似性分别为39.3%和30.3%,与人类、小鼠等哺乳类动物的相似性分别为33.3%和30.9%。nanos3基因在黄鳝肌肉中不表达,在精巢、肾脏、脾脏、脑、肝脏、肠道、心脏和血液中的相对表达量较低,在卵巢中的相对表达量最高,极显著高于在其他组织中的相对表达量(P<0.01),呈明显的组织特异性表达模式;冰冻切片荧光原位杂交分析结果显示,黄鳝nanos3基因仅在生殖细胞中表达,支持细胞中未见表达。【结论】黄鳝nanos3基因进化相对保守,主要在卵巢的生殖细胞中表达,且以细胞质中的表达为主,与核遗传信息的传递相关,是黄鳝性腺发育及分化相关的功能基因。

nanos1基因在日本血吸虫不同发育时期的定位

目的探讨nanos1基因在日本血吸虫的定位以及不同发育时期的表达情况。方法在NCBI上搜索日本血吸虫nanos1基因,PCR扩增、测序并将其全长序列经BLAST软件分析,利用DNAMAN6.0和ClustalX2.0.9进行多重序列比对,利用MAGA 5.05构建进化树进行系统进化分析。体外转录地高辛(DIG)标记的RNA探针,并将其用于日本血吸虫感染后第18、24和42天3个发育时期的雌雄虫整体原位杂交,以确定nanos1基因在日本血吸虫的定位。结果PCR扩增获得日本血吸虫nanos1基因,生物信息学分析结果表明其编码的蛋白与曼氏血吸虫nanos2蛋白同源性最高。体外转录成功的RNA探针用于日本血吸虫整体原位杂交,在感染后第24、42天雌虫的卵巢和卵黄腺以及18天雌虫的卵巢检测到阳性杂交信号,而各个发育阶段的雄虫均未观察到明显阳性杂交信号。结论日本血吸虫nanos1基因高表达于感染后第18天雌虫的卵巢以及第24、42天雌虫的卵巢和卵黄腺。该研究初步揭示了日本血吸虫nanos1基因的表达模式,为研究nanos1基因的功能提供了重要线索。

Synthesis of Nanosized NaY Zeolite by Confined Space Method

Nanosized NaY crystals have been prepared from metakaolin and sodium silicate by confined space synthesis with starch additive. It is found that the product has a narrow crystal size distribution (50-100 nm), high Si/Al ratio (Si/Al=4.6-6.1), high surface area (1090 m2/g) and the average diameter of nanosized NaY (75 nm) synthesized is 30 nm, it is smaller than that of without starch additive.

Study on Desulfurization Efficiency and Products of Ce-Doped Nanosized ZnO Desulfurizer at Ambient Temperature

Ce-doped nanosized ZnO desulfurizer was prepared by homogeneous precipitation,and its desulfurization efficiency at ambient temperature was investigated through dynamic experiments.The results showed that the desulfurization activity of nanosized Ce-ZnO had improved greatly,compared to nanosized ZnO desulfurizer.Nanosized Ce-ZnO desulfurizer was characterized by XRD,TPD-MS,XPS,and TEM.The research results indicated that doping Ce decreased the particle size of the nanosized ZnO desulfurizer and ZnS was the principal desulfurization product.There were adsorption complexes of HS and S on the surface of desulfurizer as well.Only a small amount of vapor appeared in the tail gas on the condition of meeting the precision of desulfurization.

家蚕生殖细胞相关基因nanos的克隆表达及在胚胎发育中的表达模式

【目的】探讨nanos基因在家蚕Bombyxmori胚胎发育中的表达模式,为进一步研究该基因在家蚕胚胎发育中的功能奠定基础。【方法】根据本实验室提交到GenBank中家蚕nanos的cDNA序列(登录号EF647589)设计引物,扩增出了一条长684bp的编码片段,对该片段进行了克隆和表达,亲和纯化表达的蛋白并免疫新西兰大白兔制备抗体。Western blot检测家蚕早期胚胎nanos的表达情况,荧光定量PCR检测nanos在整个家蚕胚胎发育中的表达情况。【结果】克隆并表达了一条长684bp的编码片段,得到了分子量约33kD的融合蛋白。用制备的抗血清对家蚕早期胚胎蛋白的Western blot检测表明,nanos在此阶段基本是恒定表达。荧光定量PCR结果显示刚产的卵中nanos的表达量最大,第2天开始急剧下降,此后到第10天表达量几乎没有变化。【结论】本实验克隆的nanos是家蚕中的一个同源物,该基因在家蚕胚胎发育中的表达模式与蜜蜂等有很大的不同,反映了昆虫生殖细胞形成机制的多样性。