中药大黄的综合研究XXXI．大黄蒽醌衍生物系统分离的改进方法

西宁大黄小碎片加到20％硫酸溶液和氯仿的混合液中,在水浴中回流以水解,提取,氯仿提取液相继以5％碳酸氢钾溶液,5％氢氧化钾溶液振荡提取,提取液分别以盐酸酸化,即分离得大黄酸,大黄素,芦荟大黄素及大黄酚和大黄素甲醚混合物等黄色沉淀物,后者再以硅胶柱层析分离,可得大黄酚和大黄素甲醚单体,上述五种蒽醌衍生物再经结晶即得纯品。     

大黄葸醌提取物对建鲤抗应激及生长的影响

将750尾建鲤随机分成5组,第1组为对照组,投喂基础日粮,另外4组为试验组,投喂基础日粮中分别添加0．5％、1．0％、2．0％、4．0％大黄蒽醌提取物。从2005年7月到9月连续投喂70d后,对鱼体进行高密度应激,测定其生长、应激前后血液皮质醇、血糖、溶菌酶等变化。结果表明：与对照组相比,应激前添加大黄葸醌提取物提高了鱼体增重率、特定生长率、鱼体丰满度、血液溶菌酶活性,降低了饵料系数与鱼体死亡率,但是与大黄葸醌提取物的添加水平不成线性关系,其中2．0％试验组还显著降低了血液皮质醇。高密度应激1d后,各组血液中的皮质醇、血糖、溶菌酶都有不同程度的增加,其中对照组最高,添加1．0％和2．0％大黄蒽醌提取物的试验组相对较低。应激前后各组鱼的攻毒试验表明：除应激前添加1．0％和2．0％大黄蒽醌提取物的组没有死鱼外,其它各组都有死鱼,其中对照组死亡率最高。因此添加1．0％-2．0％大黄蒽醌提取物提高了机体抗应激能力,并对病原菌感染起一定的保护作用,促进了鱼体生长。     

不同切制条件对大黄饮片中蒽醌化合物含量的影响

目的:采用高效液相色谱法比较不同的大黄切制条件对蒽醌类化合物含量的影响.方法:以大黄素和大黄酚为对照品,采用Merck C18色谱柱(4.6 mm×250 mm,5μm),甲醇-水-磷酸(体积比78.7:20.9:0.4)为流动相,检测波长254 nm,外标法测定了不同切制条件下得到的18份饮片样品中游离和结合型大黄素和大黄酚的含量.结果:从18份样品测定的平均值来看,游离型大黄素和大黄酚的含量为8.69 mg/g,结合型的为4.45 mg/g.从切制条件来看,薄片、中片、厚片总蒽醌(即游离型和结合型大黄素和大黄酚之和)含量平均分别为13.47、13.33和12.6mg/g;60℃干燥10.5 h、80℃干燥6.5 h和100℃干燥4 h,其总蒽醌含量平均分别为13.73、12.86和12.8 mg/g.结论:大黄类饮片切制的适宜条件为薄片或中片,60℃干燥10.5 h.     

HPLC测定炎消迪娜糖浆中大黄酚的含量

本文用HPLC法测定炎消迪娜儿糖浆中大黄酚的含量。色谱条件：采用Luna 5u C18柱,流动相为甲醇-水（含0．1％醋酸）（98：2,v／v）,检测波长254nm。：大黄酚在0．008～0．088μg（r=0．9997）范围内成线性关系。平均回收率（n=9）为98．79％,RSD为2．24。实验结果表明本法简便,可靠,可用于该制剂的质量控制。     

大黄素、大黄酸对平滑肌细胞增殖抑制作用研究

采用ＭＴＴ法（四氮唑法）及流式细胞技术测定大黄素、大黄酸对平滑肌细胞增殖抑制作用。结果显示,大黄素抑制平滑肌细胞增殖的有效剂量范围为其血药浓度的２５～１２５倍;大黄酸有效剂量范围为其血药浓度的１２．５～７５倍。大黄素对平滑肌细胞增殖抑制率２４．６％～９４．５８％,大黄酸抑制率为１４．３８％～８５．６４％。流式细胞技术测定大黄素处理后的平滑肌细胞出现凋亡细胞峰,Ｓ期细胞减少。     

大黄中大黄酸提取工艺研究

以大黄酸含量为参考指标,通过单因素实验分别考察硫酸浓度(A)、液料比(B)、提取加热时间(C)、提取加热温度(D)对大黄中大黄酸提取量的影响,并根据单因素试验结果设计正交试验。结果表明,影响大黄中大黄酸提取工艺的主次因素为:加热温度D>加热时间C>硫酸浓度A>液料比B,大黄中大黄酸的最佳提取工艺为:15%硫酸、5∶1液料比、55℃加热回流2.5 h。此法操作性强,优选出最佳提取工艺条件,为开发大黄中泻下有效部位提供了理论依据。     

唐古特大黄组织培养技术的研究

试验选用唐古特大黄(Rheum tunguticum Maxim．ex Regel．)种了萌发的无菌苗及无菌苗子叶、下胚轴、胚根和幼根作为材料,研究唐古特大黄不同外植体的离体培养技术。结果表明,唐古特大黄的无菌苗和无菌苗子叶、下胚轴、胚根和幼根都可以作为离体培养的良好外植体。唐古特大黄的最适分化培养基足：B5 NAA0．1mg／L 6-BA3mg／L;最适乍根培养素是：1／2MS NAA1mg／L 3%蔗糖或1／2MS NAA0．5mg／L 3％蔗糖;愈伤组织诱导培养基是：MS 2,1-D 1mg／L NAA1mg/L 6BA1mg／L。     

唐古特大黄叶柄的营养成分分析

对唐古特大黄植物地上部分基生和茎生叶柄的常规营养成分及维生素C、氨基酸、无机元素和有机酸进行了分析,结果表明,叶柄蛋白质含量较高,为5．84％,氨基酸组成全面,含量丰富;含有丰富的人体必需无机元素和较高含量的苹果酸。     

HPLC测定决明不同组织中大黄酚和橙黄决明素含量

为了测定决明不同组织中大黄酚和橙黄决明素的含量。分别收集了决明种子、根、茎、花、叶、果荚、子叶、胚轴、愈伤与不定根等组织,利用回流提取法提取大黄酚与橙黄决明素,高效液相色谱法(HPLC)进行含量测定。采用依利特反相色谱柱,以大黄酚和橙黄决明素为指标,以乙腈-0.1%磷酸水为流动相进行梯度洗脱,流速1 m L/min,检测波长为284 nm,柱温30℃。结果表明:大黄素型蒽醌类化合物在决明中的分布具有明显的组织特异性,其中,决明种子、胚轴、不定根、子叶与愈伤组织中含有大黄酚与橙黄决明素,决明根中仅含有大黄酚,其余组织不含有大黄酚与橙黄决明素,其余总蒽醌含量依次为子叶不定根愈伤组织胚轴根花。决明种子成熟过程中,大黄酚的含量呈现"波浪式上升"的趋势,峰值出现在11月下旬;而橙黄决明素的含量呈现"先上升后下降"的趋势,峰值出现在11月中旬。由于大黄酚与橙黄决明素的含量在决明不同成熟时期分别出现峰值,建议采收11月下旬的决明种子以获得大黄酚,采收11月中上旬的决明种子以获得较多的橙黄决明素。     

中药抗厌氧菌的系列研究：Ⅲ.大黄抗厌氧菌的实验研究

本研究进一步用大黄水煎液,大黄醇提物和大黄蒽醌衍生物(芦荟大黄素、大黄酸和大黄素),对临床分离的100株厌氧菌进行MIC测定,并对部分菌株进行MBC测定和亚抑菌浓度(Sub-MIC)下细菌形态观察。结果表明,大黄水煎液在1600μg/ml浓度时能抑制74％厌氧菌生长,大黄醇提物的MIC约为水煎液的1/15。三种蒽醌衍生物在8μg/ml时能抑制76～91％厌氧菌生长,这与国际公认的抗厌氧菌药甲硝唑相近。对部分菌株的MBC测定表明,大黄的MBC要大于MIC几倍以上,说明大黄抗厌氧菌主要是抑菌不是杀菌。从Sub-MIC下厌氧菌形态改变提示,大黄主要是抑制细胞壁的合成。     

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.     

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.     

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 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.