RGD/FA双靶纳米金棒的制备及其在肿瘤细胞协同靶向成像与光热治疗中的应用

目的:通过制备RGD/FA双靶纳米金考察其与高表达整合素与叶酸受体B16细胞的协同靶向成像与热疗作用;方法:采用功能化PEG分子将靶向小分子RGD与叶酸通过强健Au-S键连接至纳米金棒表面,利用激光共聚焦与808 nm近红外激光器评价修饰纳米金的协同靶向作用;结果:RGD与叶酸分子被成功连接于纳米金表面,且双靶纳米金对小鼠黑色素瘤细胞具有较好的协同靶向作用;结论:同时靶向同一肿瘤细胞的不同表位,可克服单一靶向功能化纳米粒子难以在肿瘤位点有效积累的问题,本研究为多功能纳米金棒在临床肿瘤早期诊断与光热治疗中的应用提供研究基础。     

抗新型隐球菌荚膜相关蛋白CAP10单克隆抗体的制备及鉴定

目的:以新型隐球菌荚膜相关蛋白CAP10为靶抗原,制备并鉴定特异性抗CAP10的单克隆抗体。方法:用纯化的重组CAP10免疫BALB/c小鼠,血清抗体效价达到适当水平时进行细胞融合;经多次亚克隆筛选出分泌特异性抗体的细胞株,制备单抗腹水并进行抗体效价测定及亚类鉴定。结果:获得11株能稳定分泌抗新型隐球菌荚膜相关蛋白CAP10的单克隆抗体杂交瘤细胞株,抗体效价高且抗原特异性强。结论:获得了针对新型隐球菌荚膜相关蛋白CAP10的单克隆抗体,为深入研究CAP10蛋白的功能,以及临床新型隐球菌的检测和血清型分析奠定了基础。     

新生隐球菌对角质形成细胞活力的作用研究

目的研究新生隐球菌体外对角质形成细胞活力的影响。方法将新生隐球菌父代标准株与子代荚膜缺陷株于体外分别与角质形成细胞分别共培养,同时设立热灭活的菌体、空白对照,再分别设立菌体与细胞直接接触与不接触共培养相互对照,分别作用0.5 h、1 h和2 h后,采用流式细胞仪检测隐球菌角质形成细胞的调亡率。结果随着时间延长,与空白对照组及热灭活组比较,实验组角质形成细胞的凋亡率逐渐增加。无荚膜株与父代有荚膜株比较,无荚膜株对细胞活力的影响在作用后1 h、2 h明显低于有荚膜株。2种菌株不直接接触培养使细胞的凋亡率明显下降;不直接接触的有荚膜株与热灭活的菌体之间比较差异不显著。结论虽然有荚膜株与无荚膜株隐球菌均可以使角质形成细胞活性明显降低,但荚膜可以显著增强菌体对细胞活力的影响;角质形成细胞活力的降低主要是通过与菌体接触培养后产生的,诱导细胞调亡需要菌体与细胞的直接接触。     

隐球菌感染体外血脑屏障模型的构建与应用

目的构建体外血脑屏障模型,并检测隐球菌不同菌株穿越血脑屏障的能力。方法本研究应用商品化的小鼠脑微血管内皮细胞系b END.3构建体外血脑屏障模型,并验证该模型应用于隐球菌穿越血脑屏障机制研究的可行性。通过构建模型,以非致病性的酿酒酵母作为阴性对照,比较新生隐球菌不同血清型标准株及基因缺陷株穿越体外血脑屏障能力的差异。结果跨膜电阻值(TEER)检测提示体外血脑屏障模型构建成功。检测结果显示酿酒酵母作为阴性对照穿越血脑屏障效率最低,新生隐球菌血清A型标准株H99穿越细胞屏障效率最强,血清D型标准株JEC21穿越细胞屏障效率显著低于H99。较之H99,黑色素酶缺陷株lac1裣穿越体外血脑屏障模型的效率没有显著差异;尿素酶缺陷株ure1裣效率显著下降(P0.05),约为标准株H99通过率的59.9%;荚膜缺陷株cap59裣突破体外血脑屏障模型效率最低,约为标准株H99的18%(P0.001)。结论隐球菌中枢系统感染体外模型成功构建。新生隐球菌突破血脑屏障的能力与其血清型以及荚膜、尿素酶等毒力因子的表达密切相关。     

隐球菌感染诊治专家共识

1简介 隐球菌属在真菌分类学上归人半知菌亚门、芽孢菌纲、隐球酵母目、隐球酵母科,引起人类感染的隐球菌主要是新生隐球菌和格特隐球菌。两种隐球菌的无性繁殖体均为无菌丝的单芽孢酵母样菌,在体外为无荚膜或仅有小荚膜,进入人体内后很快形成厚荚膜,有荚膜的隐球菌菌体直径明显增加,致病力明显增强。     

隐球菌感染体外血脑屏障模型的构建与应用CSCD

目的构建体外血脑屏障模型,并检测隐球菌不同菌株穿越血脑屏障的能力。方法本研究应用商品化的小鼠脑微血管内皮细胞系b END.3构建体外血脑屏障模型,并验证该模型应用于隐球菌穿越血脑屏障机制研究的可行性。通过构建模型,以非致病性的酿酒酵母作为阴性对照,比较新生隐球菌不同血清型标准株及基因缺陷株穿越体外血脑屏障能力的差异。结果跨膜电阻值(TEER)检测提示体外血脑屏障模型构建成功。检测结果显示酿酒酵母作为阴性对照穿越血脑屏障效率最低,新生隐球菌血清A型标准株H99穿越细胞屏障效率最强,血清D型标准株JEC21穿越细胞屏障效率显著低于H99。较之H99,黑色素酶缺陷株lac1裣穿越体外血脑屏障模型的效率没有显著差异;尿素酶缺陷株ure1裣效率显著下降(P<0.05),约为标准株H99通过率的59.9%;荚膜缺陷株cap59裣突破体外血脑屏障模型效率最低,约为标准株H99的18%(P<0.001)。结论隐球菌中枢系统感染体外模型成功构建。新生隐球菌突破血脑屏障的能力与其血清型以及荚膜、尿素酶等毒力因子的表达密切相关。     

新生隐球菌SER5基因的敲除与功能验证研究

目的构建新生隐球菌SER5基因缺陷株,并初步探究其功能。方法敲除SER5基因,通过体外应激应答、黑色素诱导、荚膜诱导、尿素酶测定、生长曲线测定、酵母双杂交实验考察SER5在新生隐球菌中的功能。结果成功构建SER5基因缺陷株,SER5基因缺失后对新生隐球菌的体外应激、荚膜、分解尿素及生长没有显著的影响,但新生隐球菌黑色素合成有显著的降低,通过酵母双杂交实验筛选出可能与Ser5相互作用的19种不同的蛋白编码基因。结论新生隐球菌Ser5影响新生隐球菌黑色素生成,可能与内质网蛋白、内质网稳定蛋白、VAMP7、磷脂转运蛋白、Gmt1、Vti1a等存在相关作用,提示Ser5参与新生隐球菌黑色素的合成或转运过程。     

纳米金壳介导的光热效应对骨肉瘤干细胞体外杀伤作用的研究

目的:探究纳米金壳介导的光热效应对骨肉瘤干细胞的杀伤作用。方法:采用无血清悬浮培养法富集骨肉瘤干细胞,通过实时荧光定量PCR检测所富集细胞CD133、SOX2、NANOG、OCT4 mRNA的相对表达量以进行鉴定。将纳米金壳与骨肉瘤干细胞共培养,应用波长808 nm的近红外激光器(1.5 W/cm~2)进行照射以激发光热效应,分别用CCK8法检测光热疗法对骨肉瘤干细胞的增殖抑制率,Annexin C-Fitc/PI双染法检测光热疗法对骨肉瘤干细胞凋亡的影响。结果:成功富集了高表达CD133、SOX2、NANOG、OCT4 mRNA的骨肉瘤干细胞;纳米金壳介导的光热效应显著抑制了骨肉瘤干细胞的增殖率,当纳米金壳浓度在0～250μg/mL范围内时,增殖抑制率与浓度呈正相关;细胞凋亡结果显示,纳米金壳结合近红外照射组细胞凋亡率显著高于对照组。结论:纳米金壳介导的光热效应对骨肉瘤干细胞增殖有明显抑制作用,主要诱导骨肉瘤干细胞发生中晚期凋亡。     

抗体和寡核苷酸双标记纳米金生物探针的制备及性能分析

基于纳米金粒子与抗体静电吸附作用,与硫醇修饰的寡核苷酸共价结合,建立一种新的双标记纳米金生物探针的制备方法.通过透射电镜（TEM）、紫外光谱、斑点免疫金渗滤法、免疫金银染色光镜观察法、荧光标记法等检测探针表征,及表面抗体活性情况和寡核苷酸的覆盖率,同时采用变性聚丙烯酰胺凝胶电泳（PAGE）检测寡核苷酸的存在.结果表明,纳米金粒子同时连接抗体和寡核苷酸后生物性能良好,且每个纳米金粒子（10±3）nm表面可覆盖寡核苷酸(92±20)条,双标记纳米金生物探针的制备具有简捷、稳定的特点.可作为一种新型探针应用于超微量蛋白质检测.     

新生隐球菌荚膜研究现状

新生隐球菌是一种专性需氧条件致病菌,它的细胞壁外包绕着一个多糖荚膜,是其主要毒性因子之一。荚膜主要包含两种多糖-葡萄糖醛酸木糖和半乳糖甘露聚糖,此外还有少部分的甘露糖蛋白。这些多糖分子除构成多糖荚膜外,同时也参与新生隐球菌与宿主之间的免疫反应。该文对新生隐球菌荚膜的结构、生物合成、免疫反应及针对荚膜的抗真菌治疗等方面作一综述,旨在为新生隐球菌相关疾病的研究提供新的思路。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.