备用根大鼠脊髓后角提取液中神经营养活性组分的分离及生物活性测定

制作备用根大鼠脊髓后角提取液，取其分子量大于10kD的组分进行SDS-聚丙烯酰胺凝胶电泳分析，发现手术侧和非手术侧的电泳区带数目大致相同，但手术侧样品的第四条蛋白质区带扫描的吸收峰面积百分比大于非手术侧样品的第四条区带，两者在量上可能有差别。将手术侧样品经交联葡聚精G-75凝胶层析，得到两个洗脱峰．第一峰洗脱液有促进体外培养的背根节神经元存活及其突起生长的作用，其电泳分析显示4条主带，分子量在40～80kD之间．实验结果提示部分去传入纤维支配的脊髓后角组织含有神经营养活性物质，该物质的分子量约在40～78kD之间．     

Inhibition of nitric oxide synthase reduces Sephadex-induced oedema formation in the rat lung: Dependence on intact adrenal function

In the present study we have investigated the effect of L-nitro arginine mono methyl ester (L-NAME), an inhibitor of nitric oxide (NO) synthase on Sephadex induced inflammation in the rat lung. Instillation of Sephadex into the airways induced an inflammatory reaction characterized by a long-lasting interstitial oedema, measured as an increase in lung weight, and an influx of inflammatory cells into the airways. L-NAME given s.c. prevented the increase in lung weight following Sephadex instillation. The inactive enantiomer D-NAME had no effect, nor did aminoguanidine which indicates that this effect of L-NAME was mediated by inhibition of the constitutive form of NOS. Treatment with L-NAME did not reduce an established oedema. In contrast, L-NAME tended to enhance the influx of oesinophils into the airways of Sephadex-instilled animals.L-NAME did not have any effect on the development of oedema in adrenalectomized rats or in animals where formation of glucocorticosteroids (GCS) was inhibited with metyrapone. L-NAME did not however, increase plasma levels of corticosterone. The present results indicate that, in this model, inhibition of NO-synthesis has marked anti-inflammatory effects. The underlying mechanism is complex but seems not to involve prevention of overproduction of NO.     

大鼠肝抑素纯化及其生物活性的检测

用ＳｅｐｈａｄｅｃＧ－５凝胶过滤层析法，进一步纯化具肝抑素生物活性的大鼠肝蛋白质粗提品，以分离的大鼠再生肝的肝细胞为靶细胞，体外检测各洗脱峰浓缩物对肝细胞增殖的制率结果证明，Ｅ峰浓缩物的抑制作用最强，其活性比为粗提品的２０倍，ＳＤＳ聚丙烯酰胺电泳图及蛋白质迁移率测定表明，该浓缩物的主要成分为分子量１３．５ｋＤ的多肽。本研究对大鼠肝抑素做了初步纯化，验证了该物质在肝再生中起重要调控作用的生物效应。     

纹状体源性神经营养因子的分离纯化和生物鉴定

纹状体是中脑黑质多巴胺神经元的主要靶组织，我们先前的研究曾指出，纹状体提取液能增一外培养的中脑黑质神经元的存活和发育，本研究是在此基础一步应用ＳｅｐｈａｄｅｘＧ－７５凝胶层析、高效液相色谱（ＨＰＬＣ）、反相高效液相色谱（Ｒ－ＨＰＬＣ），ＳＤＳ－ＰＡＧＥ等的分析，结合生物鉴定，从纹状体提取液中分离出化的活性因子－－纹关腐朽源性神经营养因子。该因子的分子量为１１．５ｋＤ，等电点为９．３９，它能促进体外     

二氢嘧啶酶的分离纯化与性质研究

二氢嘧啶酶产生菌Pseudomonasputida 980 1经过超声波粉碎及分级盐析 ,DEAE 纤维素和羟基磷灰石柱层析 ,SephadexG 2 0 0凝胶过滤得到纯化 2 49倍的酶液。SDS PAGE显示为单带 ,单体分子量为 380 0 0 ,SephadexG 2 0 0凝胶过滤测得分子量为 15 6 0 0 0。二氢嘧啶酶对苯海因的km 为 2 .39× 10 -2 mol/L ,其最适 pH和最适温度分别是 8.5～ 8.7和 34℃～ 35℃。 1mmol/L的EDTA和 β 巯基乙醇对酶活力有强烈的抑制作用。     

Preparation of cell-affinity adsorbents by immobilization of peptides to Sephadex G-10

In this study, affinity adsorbents for the binding of activated blood platelets and endothelial cells were prepared from Sephadex G-10 by immobilization of peptides, derived from the cell-adhesive amino acid sequence RGD (arginine-glycine-aspartic acid). Derivatization of Sephadex G-10 was accomplished by sequential coupling of specific dipeptides (RG and DV) (V = valine) and by coupling of the RGD-containing hexapeptide GRGDSP (S = serine, P = proline). Two types of gel were prepared by sequential coupling, designated as G10 (acetone) and G10 (dimethylformamide) (G10 (DMF)), containing peptides which had been coupled to the outer side of the beads and throughout the porous beads, respectively. The binding capacity of the prepared Sephadex-derivatives amounted up to 2 x 10⁹ human blood platelets per millilitre GRGDV-Sephadex at immobilized peptide concentrations, that were in the nanomole range (per millilitre packed gel) and which differed a factor 10 between G10 (acetone) and G10 (DMF). In a second series of experiments, different amounts of the hexapeptide GRGDSP were coupled to carboxylated Sephadex G-10 and carboxylated Sepharose CL 6B. The binding of human umbilical vein endothelial cells to the resulting materials was studied. Up to 10⁶ endothelial cells attached per ml GRGDSP-derivatized hydrogel at peptide concentrations of 15 nmol GRGDSP/ml Sephadex and at ±300 nmol GRGDSP/ml Sepharose. Substitution of the arginine residue of the RGD-sequence by glutamine abolished the cell-binding activity of the immobilized peptide towards activated blood platelets but not towards endothelial cells. From the results of this study it can be concluded that small peptides can be coupled to the outer side of the porous Sephadex beads, resulting in high effective ligand densities for cell-affinity applications. In this respect, Sephadex G-10, derivatized according to 'the acetone method', is a good alternative for polystyrene and other solid phase materials.     

香叶木素固体脂质纳米粒的制备及质量评价

目的 制备香叶木素固体脂质纳米粒并对其进行质量评价。方法 采用溶剂注入法制备香叶木素固体脂质纳米粒,用 Box-Benhnken效应面法优化处方,并通过包封率、微观形态、粒径分布和Zeta电位对香叶木素固体脂质纳米粒的质量进行评价。 结果 香叶木素固体脂质纳米粒最优处方组成：表面活性剂浓度3.39%,棕榈酸浓度0.116%,脂药质比为21:100,制备的香叶木素 固体脂质纳米粒外观澄清透明,带淡蓝色乳光;平均粒径为(91.73±3.18)nm(n=3),PDI为0.228,电位为(-11.46±0.74)mV(n=3);包 封率为95.13%,载药量为9.04%;透射电镜照片显示纳米粒大小均一,呈球形或类球形。 结论 该处方可用于香叶木素固体脂 质纳米粒的制备,工艺简单,稳定可行。     

The separation of total plasma insulin from binding proteins using gel filtration: Its application to the measurement of rate of insulin disappearance

Summary Insulin levels in insulin-treated diabetics were measured by a simple technique using gel filtration on Sephadex G-50 at low pH to separate plasma insulin from binding proteins. This method was used to measure the rate of disappearance of intravenously injected insulin in the plasma of diabetics with high levels of antibodies to heterologous insulin. — The half-time of disappearance of insulin was found to be closely related to insulin antibody titre, but was not related to the acute hypoglycaemic effect of the injected insulin.

---

Die Trennung von Gesamtplasma-Insulin und Bindungs-Eiwei durch Gel-Filtration: Ihre Anwendung auf die Messung der Insulinschwundrate

Zusammenfassung Die Insulinspiegel bei insulinbehandelten Diabetikern wurden mit Hilfe einer einfachen Methode unter Verwendung der Gel-Filtration auf Sephadex-G-50 bei niedrigen pH zur Trennung des Plasmainsulins und der Bindungs-Eiweißkörper gemessen. Das Verfahren diente zur Messung der Schwundrate von intravenös injiziertem Insulin aus dem Plasma von Diabetikern mit hohen Spiegeln von Antikörpern gegenüber heterologem Insulin. — Die Halbwertszeit für das Verschwinden des Insulins stand dabei in engem Zusammenhang zu dem Insulin-Antikörper-Titer, wies jedoch keine Beziehungen zu dem akuten blutzuckersenkendem Effect des injizierten Insulins auf.

---

Utilisation d'une technique de filtration pour séparer l'insuline plasmatique totale des protéines auxquelles elle est liée: son application à la mesure du taux de disparition de l'insuline

Résumé L'insulinémie chez les diabétiques traités par l'insuline a été mesurée par une technique simple utilisant le passage sur une colonne de Sephadex, à un pH bas, pour séparer l'insuline plasmatique des protéines auxquelles elle est fixée. Cette méthode a été employée pour mesurer le taux de disparition de l'insuline injectée par voie intraveineuse chez des diabétiques ayant des anticorps anti-insuline hétérologue à des taux très élevés. — La demi-vie de l'insuline ainsi déterminée est en relation étroite avec le taux d'anticorps antiinsuline mais est sans rapport avec l'effet hypoglycémique de l'insuline injectée.

     

Sephadex G10凝胶色谱法测定磺苄西林钠中的高分子杂质

目的 建立测定磺苄西林钠中高分子杂质的方法.方法 采用高效液相色谱法,色谱柱为Sephadex G10凝胶柱,流动相A为0.05 mol·L-1的磷酸盐缓冲液(pH7.0)、B为高纯水,流速为1 ml·min-1,检测波长254 nm,柱温为25℃.结果 磺苄西林钠中高分子杂质含量小于0.12%.结论 所建方法灵敏、准确、简便,可用于磺苄西林钠中高分子杂质的质量控制.     

粉尘螨抗细菌活性物质的分离与鉴定

目的从粉尘螨丙酮提取液中分离抗细菌活性成分,并对其抗菌活性进行初步鉴定。方法用丙酮抽提粉尘螨,并用Sephadex G50分子筛层析进行对粉尘螨提取液中的抗细菌活性进行初步分离、纯化。对各组分的抗细菌活性进行初步鉴定。结果经Sephadex G50分子筛层析分离得到Ⅰ及Ⅱ两个峰。粉尘螨丙酮粗提物对绿脓假单胞菌无抑制作用,但对金黄色葡萄球菌、枯草芽孢杆菌及短小芽孢杆菌均有抑制作用。峰Ⅰ及峰Ⅱ则对大肠埃希杆菌、枯草芽孢杆菌、短小芽孢杆菌及金黄色葡萄球菌都有抑制作用。粉尘螨粗提液及第二个峰在加热及蛋白酶K处理后仍然保留抗细菌活性,但是第一个峰在同样处理后失去了抗菌活性。结论首次从粉尘螨分离纯化得到抗细菌成分。