芒柄花黄素在体外对内皮细胞细胞周期的影响

目的观察芒柄花黄素在体外对人脐静脉内皮细胞（HUVEC）增殖及周期的影响。方法采用四甲基偶氮唑蓝（MTT）法检测不同芒柄花黄素对HUVEC增殖的影响,流式细胞术检测细胞周期,Western blot检测cyclin D1蛋白表达水平。结果芒柄花黄素呈剂量依赖性促进HUVEC增殖。且药物作用后,S期细胞比例增加,cyclin D1蛋白表达升高。结论芒柄花黄素对人脐静脉内皮细胞有明显的促进增殖作用,可通过上调cyclin D1蛋白表达增加S期细胞比率。     

刺芒柄花素增强脑缺血半暗带自噬流改善神经损伤

目的 刺芒柄花素（formononetin,FOR）常被用于脑卒中后的神经保护和神经功能康复,其在脑缺血再灌注（ischemia-reperfusion,I/R）损伤中的自噬溶酶体功能尚未见报道。本研究旨在探讨刺芒柄花素的治疗效果是否受到自噬流调节方式的影响。方法 雄性Sprague-Dawley（SD）大鼠在接受大脑中动脉闭塞（middle cerebral artery occlusion,MCAO）和I/R后被分为假组、模型组和MCAO+FOR（30 mg/kg）组。然后获得缺血半暗带的脑组织,分别用Beclin-1、LC3、SQSTM1/P62、Ubiquitin、LAMP-2、Cathepsin B、Cathepsin D抗体通过蛋白质印迹法（Western blot,WB）和双重免疫荧光检测自噬/溶酶体通路中的关键蛋白质。同时评估脑梗死体积、神经功能损伤和神经元坏死情况,以评价FOR的干预效应。结果 研究表明,与MCAO组相比,FOR干预除了能促进神经元中Beclin-1和LC3的自噬活性外,还能够改善自噬清除功能,如LAMP-2、Cathepsin B、Cathepsin D的溶酶体活性增强,以及Ubiquitin和P62的自噬积累减少。另外,FOR干预7 d后显著减少了脑缺血导致的神经功能缺损、梗死体积以及神经元死亡。结论 FOR的干预治疗可能是通过增强自噬流促进脑缺血半暗带恢复的神经保护机制。     

高速逆流色谱分离制备甘草中的甘草苷和芒柄花苷

应用高速逆流色谱分离制备甘草中的甘草苷和芒柄花苷。将甘草乙酸乙酯提取物经聚酰胺柱粗分后,30%乙醇洗脱物用高速逆流色谱进一步分离,所用两相溶剂系统为乙酸乙酯-水(5∶5,v/v),转速850 rpm,流速2.0 mL/min,检测波长254 nm,从50 mg30%乙醇洗脱物中得到甘草苷8.7 mg、芒柄花苷4.2 mg,纯度分别为99.5%和97.3%。所得产物的结构经核磁共振谱(NMR)鉴定。利用该方法可以对甘草中的甘草苷和芒柄花苷进行快速的分离和纯化。     

生长激素和催乳素放射免疫测定法的建立与应用

目的：建立测定大鼠垂体和血浆中生长激素（GH）和催乳素（PRL）含量的高特异性、高灵敏度的双抗放射免疫测定（RIA）法;研究急性低氧对垂体激素GH和PRL的作用。方法：用氯胺－T法进行抗原放射性碘标记;采用平衡饱和加样程序的双抗RIA法测定。结果：用该方法测定急性低氧（0．5h）时血浆和垂体GH和PRL含量,7km低氧,垂体GH含量明显升高（P＜0．05）,血浆则相反;7km低氧,明显降低垂体和血浆PRL含量（P＜0．01）;而5km低氧对GH和PRL的作用与对照组比无统计学差异。结论：本双抗RIA法具有高特异性、高灵敏度及简便易行等特性;用该法测定提示急性低氧可抑制大鼠GH和PRL的分泌。     

二氢玉米素核苷组细胞分裂素的放射免疫测定法

本文报道二氢玉米素核苷（ＤＨＺＲ）组细胞分裂素放射免疫测定法（ＲＩＡ）的研制结果。以（３Ｈ）二氢玉米素作示踪剂,测得免抗ＤＨＺＲ抗血清的效价为１：１３７０（Ｂ％＝３０％）。该抗血清主要与本组细胞分裂素发生交叉反应。回收率为９６．９％,灵敏度为１２ｆｍｏｌ ＤＨＺＲ／管,检测线性范围为０．１￣１００ｐｍｏｌＤＨＺＲ／管。批内误差ＣＶ＝４．３,批间误差ＣＶ＝２．０％。对基于同一免抗血清的ＤＨＺＲ组细胞     

RP-HPLC法测定怀槐中染料木素和芒柄花黄素含量

怀槐(Maackia amurensis Rupr.et Maxim.)广泛分布于中国东北三省及内蒙古等地,尤以黑龙江省为多.怀槐作为民间药,其茎、枝可治疗风湿性关节炎; 叶、枝皮可治疗肿瘤; 内皮可治疗深度伤口及久不愈合的溃疡.另外,怀槐树皮在民间还作为祛风湿、消炎、镇痛、健胃、止血等药广泛应用,疗效甚佳.从怀槐的根、枝、叶、果实和心材中分别分得异黄酮、生物碱、NC442类化合物,据报道,其中的异黄酮类化合物表现出对心血管系统的复合作用和明显的抗肿瘤活性[1].作者在对怀槐进行系统的化学成分研究过程中,分离鉴定了9个异黄酮类化合物[2,3],并进行了抑制肿瘤细胞增殖作用研究[4],其中,染料木素(genistein)对胃腺癌细胞(BGC)增殖有一定程度的抑制作用,芒柄花黄素(formononetin)则对白血病细胞(HL-60)增殖有一定的抑制作用.染料木素和芒柄花黄素作为马鞍树属(Maackia Rupr.)植物中普遍存在的2个异黄酮类化合物,其活性研究日益受到重视.采用HPLC法测定染料木素等异黄酮类化合物含量在大豆等其他植物中曾有文献报道[5].为了尽早开发怀槐资源,同时为其资源的综合利用提供科学依据,本文采用HPLC法对怀槐不同部位的染料木素和芒柄花黄素2个有效成分进行了含量测定.     

棒柄花的化学成分研究

从棒柄花(Cleidion brevipetiolatum)的乙醇提取物中分离得到6个化合物,经波谱分析确定其结构为：benzyl-1-O-β-D-glumpyranoside(1),chavicol-1-O-β-D-glumpyranoside(2),3,4,5-trimethoxyphenol(3),3,4,5-trimethoxyphenol-1-O-β-D-glucopyranoside(4),β-sitosterol(5),daumsterol(6),以上化合物均为首次从该植物中分离得到。     

红柄白鹃梅营养成分的分析

对红柄白鹃梅嫩茎、幼叶和花蕾等器官不同部位的粗脂肪、可溶性蛋白、粗纤维、可溶性糖、维生素C的含量进行了分析测定.结果表明红柄白鹃梅粗纤维含量丰富,达4.45g.100g-1,可溶性糖的含量较低为2.16g.100-g1,粗脂肪和维生素C的含量适中,分别为0.69g.100g-1和32mg.100-g1,可溶性蛋白质的含量较低,为0.317g.100g-1.红柄白鹃梅富含营养成分,是一种极具开发利用价值的木本野生蔬菜.     

双峰驼血浆促卵泡素的放射免疫分析法

用单峰驼促卵泡素标准品（ＣａｍＦＳＨ）,ｈＦＳＨ抗血清和＾１２５Ｉ－ｈＦＳＨ建立了测定双峰驼血浆ＦＳＨ的放射免疫分析方法,并通过一系列实验证明,该方法可以用于测定双峰驼血浆ＦＳＨ,是研究双峰驼生殖内分泌学的可靠手段之一。     

百合科扭柄花属一个新的染色体基数

对中国云南西部和西北部分布的腋花扭柄花Streptopus simplex的4个居群进行了细胞学研究。生长在云南西北香格里拉县(原中甸县)碧塔海和小中甸冷杉林中的腋花扭柄花两个居群的体细胞染色体数目为2n=2x=18,而生长在高黎贡山的福贡县片马和贡山县的灌丛中的植物体细胞染色体数目则为2n=2x=14。2n=14为腋花扭柄花一个新的染色体数目,x=7为扭柄花属一个新的染色体基数。香格里拉碧塔海和小中甸两个居群的核型公式分别为2n=4m+8sm+4st和2n=8m+2sm+6st,染色体逐渐变小;贡山和福贡片马两个居群的核型公式分别为2n=14=4m+10sm和2n=14=7m+7sm,其中第一对中部着丝粒的染色体显著大于其余染色体。由于x=8是扭柄花属最常见的染色体基数,因此可认为x=8是腋花扭柄花的染色体原始基数,x=7的数目是衍生的;x=7居群染色体的一条大染色体可能是由x=8的染色体的两条st型染色体的着丝粒发生了罗伯逊易位而来。     

Radioimmunoassay determinations of decreased amounts of α-L-fucosidase protein in fucosidosis

Purified human liver α-l-fucosidase (EC 3.2.1.51) has been radioiodinated by a chloramine-T procedure to a specific activity of 3.7·10⁶ dpm/μg protein without altering its apparent Michaelis constant for the 4-methylumbelliferyl substrate. This ¹²⁵I-labelled α-l-fucosidase has been used in development of a competitive binding radioimmunoassay for α-l-fucosidase which can detect 1–2 ng of enzyme protein and has been employed to quantify the amount of α-l-fucosidase protein in the liver and spleen from a patient with fucosidosis. Less than 1% of the normal amount of α-l-fucosidase protein is present suggesting that normal amounts of catalytically inactive α-l-fucosidase are not found in this disease.     

Development and Applications of a Solid-Phase Radioimmunoassay for the P0 Protein of Peripheral Myelin

This is the first report of a quantitative radioimmunoassay for PO. The assay uses antigen-coated plastic microwells, with antibody binding detected by 125I-labeled protein A. Either peripheral myelin proteins or purified PO may be used as the antigen. Optimal extraction of tissue samples for PO immunoassay requires careful attention to the sodium dodecyl sulfate-to-protein ratio. Sodium dodecyl sulfate interference with antibody binding can be minimized by adding an excess of nonionic detergent and carrier protein to the incubation buffer. This method allows the detection of 0.8 ng of PO (20 ng/ml). Results from this assay showed little or no immunoreactivity in extracts of brain, centra myelin, liver, purified myelin basic proteins, cultured, purified secondary Schwann cells, or membrane preparations from these cells. PO was clearly detectable in Schwann cell cultures from 3- to 4-day-old rats at 12-18 h after dissociation (4% of the level in adult sciatic nerve) and in extracts of one-day-old rat sciatic nerve (2% of the level in adult nerve). Myelin basic protein radioimmunoassays showed that the ratio of PO to myelin basic protein is essentially constant in extracts of sciatic nerve from ne-day-old, four-day-old, and young adult rats. Another result was that PO levels are reduced in the trembler mouse sciatic nerve.     

Radioimmunoassay of digitonin

We describe a radioimmunoassay for digitonin which utilizes the ability of digotonin to compete with ¹²⁵I-labeled digoxigenin for binding to anti-digoxin antiserum. As performed using the Gammaflo automated radioimmunoassay system and commercially available reagents, the assay can detect as little as 20 μg/ml (15 μM) digitonin. The assay is insensitive to interference by cholesterol or other cell membrane constituents and is useable above and below the critical micelle concentration of digitonin. It should be useful for the monitoring of digitonin concentrations in solubilized biochemical preparations.     

Rhizobium trifolii mutants deficient in exopolysaccharide production

Spontaneous mutants of Rhizobium trifolii 24AR5 which did not produce exopoly-saccharide were isolated. The non-mucoid mutants formed small white and ineffective nodules on both red and white clover. These nodules contained infection threads, but only a small number of bacteria were released into nodule cells, and bacteroids were rarely observed. The non-mucoid phenotype was not complemented by the symbiotic plasmid (pJB5JI) of Rhizobium leguminosarum.     

Comparative structural analysis of snail galactans by a radioimmunoassay to elucidate species-specific determinants

Summary Galactans, the storage polysaccharides in the perivitelline fluid of many snails showed a high degree of species-specificity as revealed by quantitative precipitin formations with lectins, polyclonal antisera, myeloma proteins as well as by the reactivity with the enzyme galactose oxidase. However, their chemical compositions were remarkably similar since thed-Gal residues were all linked 13 and 16 glycosidically.The specificity seemed to be related to the different degrees of branching in the various galactans but could also be due to some other minor constituents in some galactans such asl-galactose or phosphate.In this study a Radioimmunoassay was developed using the galactan of the snailLymnaea stagnalis to elucidate those differences which were only related to a unique distribution of the 13 and 16 linkages, since this galactan was composed exclusively ofd-galactose residues. The galactan was labeled by sequential oxidation with galactose oxidase and reduction with tritiated sodium borohydride. Inhibition of the binding of the labeled galactan to insolubilized antibodies was investigated by galactans of different species, their chemically modified products, andd-galactose-composed oligosaccharides of unambiguously identified structures.Inhibition byLymnaea stagnalis galactan was about 45 000 times that ofHelix pomatia galactan. The most complementary oligosaccharide found was -d-Gal13[-d-Gal16]-d-Gal11l-Gro, being about 200 times more effective thand-Gal. However, a fraction with molecular weights between 700 and 1000 isolated from the partially hydrolized galactan was still seven times more effective. From the reactivity of the antiserum with the different oligosaccharides tested the following structure was inferred which most likely represented the complete determinant recognized by the antiserum: -d-Gal13[-d-Gal16]-d-Gal16[-d-Gal13]-d-Gal1. This determinant seemed to be most common inLymnaea stagnalis galactan and its frequency of occurrence appears to correspond to the inhibitory potency of other snail galactans.