匹伐他汀对体外培养人脐静脉血内皮祖细胞数量及功能的影响

目的：通过体外培养人脐静脉血内皮祖细胞（endothelial progenitor cells,EPCs）,观察他汀类新药（匹伐他汀）对EPCs数量及增殖、迁移和粘附功能的影响。方法：采用密度梯度离心法分离培养人脐静脉血单个核细胞,将其接种在包被有人纤维连接蛋白培养板上,培养7 d后,收集贴壁细胞,加入不同浓度匹伐他汀（分别为0.001 μmol/L、0.01 μmol/L、0.1 μmol/L、1.0 μmol/L）培养24 h,用免疫荧光法观察EPCs 吸收FITC-UEA-I 和Dil-acLDL情况对EPCs 进行鉴定,然后分别采用MTT 比色法、改良的Boyden小室、粘附能力测定实验对各实验组测定,来观察匹伐他汀对EPCs 数量及增殖、迁移和粘附功能影响。结果：匹伐他汀组与对照组相比,匹伐他汀显著提高了体外培养EPCs的数量及增殖、迁移与粘附能力（P〈0.05）。匹伐他汀浓度在0.1 μmol/L 时对EPCs影响达到最大。随着药物浓度的继续增大,EPCs的上述功能反呈下降趋势,但1.0 μmol/L 组仍高于对照组。结论：匹伐他汀能增加体外培养EPCs的数量及增殖、迁移和粘附能力,可作为EPCs 培养的一种改良方法,为其更好的应用于临床具有重要的意义。     

他汀类药物对外周血内皮祖细胞的影响

本文旨在探讨他汀类药物氟伐他汀对外周血内皮祖细胞(endothelial progenitor cells,EPCs)数量和功能的影响.用密度梯度离心从外周血获取单个核细胞,将其接种在人纤维连接蛋白(human fibronectin)包被的培养板中,培养7 d后,收集贴壁细胞,加入不同浓度氟伐他汀(分别为0.01、0.1、1、10μmol/L)和辛伐他汀(1 μmol/L),培养一定的时间(6、12、24、48 h).用激光共聚焦显微镜鉴定FITC-UEA-I和DiI-acLDL双染色阳性细胞为正在分化的EPCs,用流式细胞仪检测其表面标志进一步鉴定EPCs,在倒置荧光显微镜下计数.采用MTT比色法、改良的Boyden小室、粘附能力测定实验和体外血管生成试剂盒观察EPCs的增殖能力、迁移能力、粘附能力和体外血管生成能力.结果显示,氟伐他汀可显著增加外周血EPCs的数量,并且EPCs数量随氟伐他汀浓度增加及作用时间延长而增加,1 μmol/L浓度氟伐他汀作用24h对EPCs的数量影响最为显著(较对照组增加15倍,P＜0.05).在动物实验中,喂养氟伐他汀3周后,大鼠的EPCs也较对照组增加2倍(P＜0.05),进一步支持了体外实验的结果.氟伐他汀和辛伐他汀也显著改善外周血EPCs的粘附能力、迁移能力、增殖能力和体外血管生成的能力,相同浓度的氟伐他汀和辛伐他汀(1 μmol/L)对EPCs数量和功能的影响并无显著差异.上述观察结果提示他汀类药物可增加EPCs的数量,改善EPCs功能.     

他汀对EPCs增殖力影响的PI3和ERK信号通道机制研究

目的:观察他汀对冠心病患者内皮祖细胞(EPCs)增殖力的影响及与PI3/Akt和ERK信号通道的相关性.方法:冠心病和非冠心病患者各16例纳入实验,非冠心病患者的10mL外周血来源的单个核细胞纳入正常组,冠心病患者的40mL外周血来源的单个核细胞均分纳入10μmol/L阿托伐他汀组、10 μmol/L阿托伐他汀+PI3/Akt通道阻滞剂LY294002组和10μmol/L阿托伐他汀+ ERK信号通道阻滞剂PD98059组,均向EPCs方向分化,以VEGFR2、CD34和AC133流式鉴定;观察冠心病患者EPCs增殖力的变化及他汀的影响,观察LY294002和PD98059分别阻断PI3/Akt和ERK通道后他汀对冠心病患者EPCs增殖力作用的变化.结果:与非冠心病人对比,冠心病患者EPCs的增殖(0.23± 0.02 to 0.14± 0.02,P＜0.001)功能下降,阿托伐他汀明显提高冠心病患者EPCs的增殖力(0.14± 0.02 to 0.20± 0.02,P＜0.05);该作用可为Pl3/Akt通道阻滞剂LY294002阻断(0.20± 0.02 t0 0.16±0.02,P＜0.001),但ERK信号通道阻滞剂PD98059无此作用(0.20±0.02比0.20±0.02,P＞0.05).结论:阿托伐他汀可通过PI3/Akt通道而非ERK信号通道上调冠心病患者外周血来源EPCs的增殖力.     

血脂康胶囊对高脂模型大鼠EPCs功能的影响

目的:高脂血症可增加心血管事件的发生率,本研究降脂红曲制剂血脂康胶囊对高脂模型大鼠的内皮祖细胞(endothelial progenitor cells,EPCs)生物学功能的影响。方法:给予Wistar大鼠高脂饲料30 d,造成高血脂大鼠模型,灌服血脂康胶囊。密度梯度离心法分别分离正常对照组,高脂模型组和血脂康治疗组大鼠骨髓单核细胞,应用EGM-2MV进行体外培养。以4~6代EPCs为靶细胞。采用Edu标记技术、CCK-8检测法、粘附能力测定试验、改良的Boyden小室、Matrigel法、荧光定量RT-PCR等方法分别检测EPCs增殖、粘附、迁移、体外成血管及单核细胞趋化蛋白-1(monocyte chemoattractant protein-1,MCP-1)等炎性因子的表达。结果:高血脂模型组大鼠EPCs的增殖、粘附、迁移及成血管能力均明显低于对照组,但炎症因子MCP-1表达则高于对照组;与高脂模型组EPCs比较,血脂康可明显促进EPCs的增殖、粘附、迁移及成血管,下调MCP-1的表达。结论:高脂状态下大鼠EPCs的增殖、粘附、迁移及成血管等生物学功能受损,血脂康能调整脂质代谢,改善高脂血症大鼠EPCs功能,从而起到保护血管内皮的功能,减少心血管疾病的发生。     

消渴丸对GK糖尿病大鼠内皮祖细胞功能的影响

目的:观察消渴丸对GK大鼠外周血中内皮祖细胞(endothelial progenitor cells,EPCs)数量及功能的影响。方法:消渴丸连续给药12周后测定并比较GK大鼠血清EPCs的粘附能力、迁移能力和体外生成血管能力。结果:消渴丸可使GK大鼠EPCs数量显著升高,其粘附能力、迁移能力和体外生成血管能力也明显改善。结论:消渴丸可以升高GK大鼠动脉血中EPCs数量并改善其功能。     

阿托伐他汀对冠心病患者循环内皮祖细胞数量与分化相关功能的影响

目的:观察冠心病患者循环内皮祖细胞数量与分化相关功能的变化特点,探讨体外条件下不同浓度阿托伐他汀对冠心病患者循环内皮祖细胞数量和增值,黏附,迁移等分化相关功能的影响。方法:以16例条件齐同的非冠心病患者作对照,观察16例冠心病患者循环内皮祖细胞数量与功能的变化,体外条件下不同浓度阿托伐他汀处理冠心病患者循环内皮祖细胞。结果:冠心病患者外周血内皮祖细胞的数量与功能下降,阿托伐他汀呈浓度依赖性提高冠心病患者内皮祖细胞的数量与增值,黏附,迁移等分化相关功能;阿托伐他汀对内皮祖细胞分化相关功能的改善作用大于对其数量的影响。结论:阿托伐他汀可上调冠心病患者循环内皮祖细胞的数量与分化相关功能,且以改善功能为主。     

补骨脂素对大鼠成骨细胞增殖与分化的影响

为探讨补骨脂素体外对大鼠成骨细胞增殖与分化的影响,用改良的组织块法分离培养新生大鼠颅骨成骨细胞,补骨脂素以不同浓度加入细胞培养体系,作用不同时间后,用MTT法检测成骨细胞的增殖情况;用对硝基苯二钠基质动力学法测定细胞内碱性磷酸酶的活性,用改良的Lowry法测蛋白含量。补骨脂素浓度在1μmol/L 24 h,5~20μmol/L浓度范围内48 h,1、10、20μmol/L浓度范围内72 h促进成骨细胞增殖,在10~15μmol/L范围内48 h及72 h提高成骨细胞内碱性磷酸酶的活性。补骨脂素体外能促进成骨细胞的增殖与分化。     

脑心清片对缺血性心脏病患者外周血中内皮祖细胞功能的影响

目的:观察脑心清片对缺血性心脏病(ischemic heart disease,IHD)患者外周血中内皮祖细胞(endothelial progenitor cells,EPCs)数量及功能的变化。方法:将40例IHD患者随机分为脑心清组30例和贝那普利组10例;用密度梯度离心法从外周血中分离单核细胞;分别观察治疗前后EPCs的粘附能力、迁移能力和体外生成血管能力。结果:脑心清组在治疗后IHD患者外周血中EPCs数量显著升高,其粘附能力、迁移能力和体外生成血管能力也明显改善。结论:脑心清片可以升高IHD患者外周血中EPCs数量并改善其功能。     

吴茱萸碱抑制人结肠癌HCT-116细胞体外增殖和迁移

吴茱萸碱(Evodiamine,Evo)对人结肠癌HCT-116细胞体外增殖和转移的影响及其可能机制初探。体外培养人结肠癌HCT-116细胞株,加入Evo1.5μmol/L、3μmol/L、6μmol/L、12μmol/L、24μmol/L、48μmol/L、96μmol/L继续培养48 h,CCK-8法检测Evo对HCT-116细胞生存活性的影响;Transwell法和划痕法检测STF-118804、Evo对HCT-116细胞体外迁移的影响;RT-PCR和Western blotting检测不同浓度的Evo(0μmol/L,1.5μmol/L,3μmol/L,6μmol/L)和STF-118804(10μg/L)诱导HCT-116细胞48 h后,细胞中SIRT1、NF-кB、MMP-9蛋白的表达情况。不同浓度的Evo(1.5μmol/L,3μmol/L,6μmol/L,12μmol/L,24μmol/L,48μmol/L,96μmol/L)作用于HCT-116细胞48 h后,细胞的增殖均受到明显抑制(p0.05),且在一定范围内呈浓度依赖性;且Evo能抑制HCT-116细胞的迁移。Evo可明显下调HCT-116细胞内SIRT1、MMP-9 m RNA和蛋白的表达,上调NF-кB m RNA和蛋白的表达。体外实验表明,Evo对结肠癌细胞增殖和迁移的抑制作用是通过抑制SIRT1的表达,激活NF-кB信号通路,进而抑制MMP-9来实现的。     

地骨皮提取液对内皮祖细胞功能的影响

通过观察地骨皮提取液对高糖培养的人脐血内皮祖细胞(EPCs)的黏附、迁移、增殖等能力的影响,探讨地骨皮提取液对高糖所致的血管内皮损伤是否具有保护作用。分离人脐血单个核细胞,接种培养后收集贴壁细胞,采用双荧光染色法及ecNOS和Flk-1基因的表达对EPCs的生物学特征进行鉴定。将分离到的EPCs分成5组:正常对照NG组、高糖HG组、HG+地骨皮提取液不同浓度组(1 g/组、2 g/L组和4 g/L组);用重贴壁法测定EPCs黏附能力,改良Boyden小室法测定其迁移能力及CCK-8法测定增殖能力。结果显示:(1)HG组EPCs的黏附、迁移及增殖能力较NG组明显下降;(2)HG+地骨皮提取液不同浓度组EPCs黏附、迁移和增殖能力均比HG组高;对迁移和增殖能力的影响以2 g/L干预组效果最为明显。提示地骨皮提取液能部分恢复高糖对EPCs黏附、迁移及增殖能力的抑制,对高糖所致的血管内皮损伤具有保护作用。     

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.     

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.     

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。     

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.     

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.     

医疗机构合理用药评价模型的构建研究

目的 针对医疗机构的合理用药水平进行评价研究。方法 根据医疗机构合理用药的具体要求,构建医疗机构合理用药评价指标体系,采用基于模糊群决策的方法和多指标评价分析法构建医疗机构合理用药评价模型。结果 构建了基于模糊群决策的医疗机构合理用药评价模型,并通过实例分析证明了评价模型的可行性。结论 建立的基于模糊群决策的医疗机构合理用药评价模型能够对医疗机构的合理用药水平进行科学评价,为提高医疗机构合理用药水平奠定基础。     

The mechanism of hatching of eggs of Haemonchus contortus

Fluid collected from hatching eggs of Haemonchus contortus contained a lipase which hydrolysed 2-naphthyl laurate (about 0·7 μmol naphthol freed /h/10⁶ eggs). The fluid also hydrolysed l-leucinamide (about 2·3 μmol leucine freed/h/10⁶ eggs). The fluid when added to normal or heated eggs caused ‘hatching’. ‘Hatching’ also occurred in exsheathing fluid from infective juveniles and in a preparation of pancreatic lipase containing leucine aminopeptidase. A purified mammalian leucine aminopeptidase in combination with several different lipases did not attack egg shells.The ‘spontaneous’ hatching of eggs of H. contortus was strongly inhibited by 1,10-phenanthroline, 10^?3M, and this inhibition was reversed by Zn²⁺. However, the inhibition of ‘hatching’ of eggs in externally applied hatching fluid, or the hydrolysis of leucinamide in hatching fluid was generally less marked.