僵蚕药材蛋白质分级指纹图谱建立及产地鉴定相关分子研究

建立云南与四川僵蚕药材的总蛋白质指纹图谱及分级指纹图谱,对比分析以发现其中的差异表达蛋白质分子并进行鉴定及生物信息学分析,探索其用于产地鉴定的可行性。提取两种僵蚕的酸溶、碱溶、水溶及醇溶性总蛋白质,对其进行丙酮分级沉淀,对所得样品进行SDS-PAGE指纹图谱对比分析,对所得差异条带进行LC-MS/MS分析,数据库检索后进行生物信息学分析。两种僵蚕的总蛋白质指纹图谱相似度很高,难以据此进行产地鉴定;两种僵蚕的分级指纹图谱则有较显著差异,有望用于产地鉴定;依据分级指纹图谱差异共鉴定出β-葡萄糖苷酶、抗胰凝乳蛋白酶、抗胰蛋白酶、含脂肪酶结构域蛋白等273种蛋白质。云南与四川僵蚕的蛋白质组成及其分级指纹图谱存在显著差异,有望用作产地鉴定的分子依据;醇溶蛋白质构建分级指纹图谱可以提供更加丰富的信息及更好的分辨率;分级指纹图谱技术具有简便易行、分辨率较高的优点,为其他中药材的产地鉴定研究提供了方法学思路。     

工业废羊毛酸解工艺条件选择

毛发蛋白盐酸水解提取胱氨酸时 ,温度、毛酸比、时间等因素影响到毛发蛋白的水解率和产品胱氨酸的总收率。通过正交试验法 ,进行了废羊毛酸解工艺参数的试验研究 ,确定了最佳酸解工艺参数为 :温度 1 0 5℃ ,毛酸比(W/V) 1∶1 .7,连续水解时间 7.0h。     

蹄叶槖吾叶多糖提取工艺优化及单糖组成研究

采用水提醇沉法提取蹄叶槖吾叶粗多糖,通过单因素试验和Box-Benhnken法优化该多糖的提取工艺。结果表明:蹄叶槖吾叶粗多糖的最佳提取工艺为:料液比1∶20(g/m L)、提取温度90℃、提取时间2 h、提取3次,提取率为8.76%;通过红外光谱扫描检测其特征基团,并应用高效液相色谱技术对已脱除蛋白的多糖结构进行初步鉴定,得到其单糖组成为半乳糖醛酸∶鼠李糖∶半乳糖∶阿拉伯糖∶甘露糖∶葡萄糖∶葡萄糖酸∶岩藻糖=43.5∶6.0∶23.6∶11.5∶6.3∶5.4∶1.6∶0.9。     

贝壳的有机质及生物矿化机制分析

以腹足纲贝壳香螺壳为研究对象,用弱酸去钙法进行蛋白提取,采用280纳米(A280)光吸收法测定蛋白含量,并通过聚丙烯酰胺(SDS-PAGE)凝胶电泳法对蛋白按照分子量大小的区别进行分离.实验结果表明香螺壳中蛋白含量和种类较少,其文石层比方解石层蛋白含量高的多,总量分别为0.89%和0.0533%;文石层分离出五种分子量的可溶性蛋白和四种分子量的不可溶性蛋白;而方解石层中分离出三种分子量的可溶性蛋白和三种分子量的不可溶性蛋白,且分子量不相同.正是这少量的蛋白质对贝壳的生物矿化过程和不同晶型的形成起着决定性作用.     

白芷醇溶蛋白质提取工艺优化及其指纹图谱研究

目的：基于响应面设计优化白芷醇溶蛋白质提取工艺，并研究其指纹图谱特征。方法：首先考察液料比、提取时间、乙醇浓度、提取温度四个单因素对白芷醇溶蛋白质提取率的影响，再通过Box-Behnken方法对影响提取率的工艺因素进行响应面优化，最后对白芷及易混淆药材的醇溶蛋白质指纹图谱进行对比分析。结果：最佳提取工艺条件为：液料比13∶1(mL/g)，提取时间8.75 h，乙醇浓度42%，提取温度45℃，在此条件下提取率为0.294%。SDS-PAGE分析表明白芷醇溶蛋白质样品分子量范围介于15～95 kDa,91.4 kDa与65.6 kDa分子为高丰度蛋白质。此外，白芷与天花粉、粉葛、山药的醇溶蛋白质指纹图谱有显著差异。结论：利用响应面法优化了白芷醇溶蛋白质的提取工艺，并建立其专属性SDS-PAGE指纹图谱，为其分子鉴定及资源开发提供实验依据。     

苦荞麦黄酮和蛋白质提取工艺的优化

采用HPLC和考马斯亮蓝检测方法,通过单因素试验结合正交试验对苦荞麦黄酮和蛋白质分步提取工艺进行优化,探寻一种高效的苦荞黄酮和蛋白质的提取工艺。结果表明,苦荞壳、苦荞麸皮黄酮提取的最优工艺条件为按料液比1∶10(g/m L)加入75%乙醇溶液,在提取温度85℃下提取2 h,提取效率为3.85%;苦荞麦心粉蛋白提取的最优工艺条件为按料液比1∶15(g/m L)加入p H=9的碱水,在提取温度35℃下搅拌提取4 h,提取效率为6.22%。优化后的苦荞麦黄酮和蛋白分步提取工艺操作简单、能耗低、较大限度地提高了苦荞麦的综合利用率。为企业进行高产、低耗、环保的连续自动化生产奠定基础。     

花椒籽黑色素提取和脱蛋白技术研究

采用二次回归正交旋转组合试验优化了花椒籽黑色素的浸提工艺,并用蛋白酶酶解-Sevag法联用脱除花椒籽黑色素中的蛋白.结果表明:影响花椒籽黑色素提取效果的因素依次为温度>NaOH浓度>料液比,花椒籽黑色素提取的优化工艺参数为NaOH浓度为1.20 mol/L,料液比为1∶24.64,温度为70℃下提取2 h,共2次,所得花椒籽色素粗品为棕黑色无定型粉末,得率为6.1%,色价为201.62,蛋白质含量为44.83%~48.63%.碱性蛋白酶脱蛋白条件为温度50℃,pH 9,加酶量为粗色素溶液(浓度为1 mg/mL)体积的6%,再用Sevag法处理3次,蛋白质脱除率可达81.23%,花椒籽黑色素色价可提高1.5倍.     

四种主要长春花生物碱超声波法提取工艺优化研究

为探索高效、低污染的长春花中文多灵、长春质碱、长春新碱和长春碱的提取工艺,进行了超声波-甲醇法提取工艺条件的优化研究。结果表明:在单因素试验基础上,通过正交试验表明,超声波-甲醇法提取文多灵和长春质碱的最佳工艺条件为:提取溶剂:甲醇(pH4.0);料液比:1∶15;提取温度:45℃,提取时间:180 min,提取频率:59 KHz;文多灵和长春质碱平均得率为0.16%,0.08%。     

美味牛肝菌多糖最佳提取工艺研究

对美味牛肝菌 (BoletusedulisBull)子实体多糖提取最佳工艺条件进行了研究。通过实验既探讨了温度、pH值、时间、料液比及浸提级数对提取效果的影响 ,也探讨了酶法、三氯乙酸—正丁醇法及复合法去除蛋白的工艺条件。结果表明 :蛋白质去除最佳工艺为酶法和三氯乙酸—正丁醇复合法 ,美味牛肝菌多糖提取最佳工艺为加水 2 0倍 ,80℃ ,pH 8时浸提 1h ,多糖浸提率可达 7.37%。     

贵州野生甜藤多糖的提取与脱蛋白方法研究

以贵州野生甜藤干粉为原料,通过单因素试验,再通过正交试验优化甜藤多糖提取工艺,并采用Sevag法、TCA法、Sevag-TCA法、HCl O4法进行脱蛋白方法研究。结果表明,甜藤多糖最优提取工艺为:以水为溶剂,料液比1∶50,超声功率70 W,水浴温度70℃,超声提取时间110 min,提取2次;HCl O4法脱蛋白效果最佳,经3次脱蛋白处理后蛋白脱出率为94.78%,多糖损失率为15.14%,该法用于脱除甜藤植物多糖中的蛋白质,工艺简单,成本低,具有一定应用价值。     

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.