用N-(3-P)NEM标记毛发角蛋白研究毛发中低硫蛋白和高硫蛋白的状态

本文用一种特异性标记蛋白质巯基的N-(3-P)NEM为荧光探针标记毛发角蛋白,经SDS-PAGE后,通过荧光薄层扫描确定N-(3-P)NEM与含巯基蛋白结合的部位。结果表明,毛发角蛋白中的低硫蛋白分子量在45,000～63,000范围,约占总蛋白量的49.6％～68.2％;高硫蛋白分子量在4,000～20,000范围,约占总蛋白量的18.9％～36.2％。除低硫和高硫蛋白以外,毛发角蛋白中还可能存在一种极低硫蛋白,分子量位于22,000～32,000范围。对人与5种动物的毛发角蛋白组分进行比较,证实人与动物的毛发角蛋白的主要差异在高硫蛋白,不同种属的动物毛发角蛋白中低硫蛋白和高硫蛋白的含量也不相同。作者认为毛发角蛋白中含巯基蛋白的差异可为动物分类学和法医学在进行毛发比较鉴别方面提供新的重要信息。     

高核酸高蛋白酵母选育及发酵生态学研究

通过紫外线嘧啶复合处理选育到一株高核酸高蛋白酵母，并进行了营子及其它培养条件的选择性试验，确定了优化的培养条件，摇瓶培养核酸１４．７５％，蛋白质含量达６３．６７％。１．５Ｌ发酵罐发酵试验能很好地重现摇瓶效果及冰箱保存活试验证明该菌株有好的遗传稳定性。     

葛根素治疗不稳定性心绞痛71例临床观察

目的探讨在常规治疗基础上应用葛根素注射液治疗不稳定性心绞痛(UA)的疗效。方法对于入院确诊为不稳定性心绞痛的患者,随机分为两组:对照组为常规抗凝组,低分子肝素5000u皮下注射,1次/12h,共5d,阿斯匹林肠溶片0.3g,1次/d,3d后改为0.1g,1次/d,长期服用;治疗组为葛根素组,葛根素300mg静脉滴注,1次/d,其余治疗同对照组。结果治疗组控制症状及改善异常心电图方面均优于对照组,并使C-反应蛋白(CRP)浓度更快下降,与对照组相比,差异有显著性(P<0.05)。结论在抗凝及抗血小板基础上加用葛根素,治疗UA疗效优于常规抗凝治疗。     

乳腺癌PKCs表达与其分化程度的相关性

为了研究蛋白激酶C（PKC）和乳腺癌分化及转移的相关性，采用免疫组化SABC方法检测48例人乳腺癌的PKC蛋白表达，包括PKCα、βI、η和ξ等蛋白。结果表明，与分化低的乳腺癌对比，分化高的乳腺癌PKC蛋白表达水平提高。在临床I－II和III级的乳腺癌中，PKCη蛋白表达率分别为61．5％和9．1％，具有显著意义（P＜0．05），PKCξ蛋白表达率分别为92．3％和27．3％，具有极显著意义（P＜0．01）。另外，在乳腺癌未转移的病例中，PKCs蛋白表达水平普遍较高，其中PKCξ与肿转移相关性极显著（P＜0．01）。此外，PKCs蛋白表达水平与肿瘤患者年龄无关。以上结果提示，PKCη和PKCξ有可能作为评价临床乳腺癌分化和转移程度的指标之一。     

Changes in glycosaminoglycan sulfation and protein kinase C subcellular distribution during differentiation of the human colon tumor cell line Caco-2

Summary During the spontaneous differentiation (day 5 to day 15 of the culture) of Caco-2 cells, the sulfation of cell layer glycosaminoglycans increased, whereas protein kinase C activity was concomitantly redistributed from the membrane to the cytosol. The protein kinase C activators, 4-phorbol 12-myristate, 13-acetate and 1,2-dioctanoyl-glycerol inhibited glycosaminoglycan sulfation. By contrast, 4-phorbol 12, 13 didecanoate was ineffective.These results suggest that membrane-bound PKC may exert a modulatory effect on glycosaminoglycan sulfation, and this effect is gradually attenuated as Caco-2 cell differentiation progresses.     

Effect of protein kinase C activation and depletion on insulin stimulation of glycogen synthesis in cultured hepatoma cells

Summary Insulin stimulation of glycogen synthesis was nearly abolished in hepatoma cells shortly treated with 4 ß-phorbol 12 \-myristate, 13 -acetate (protein kinase C activation) but remained unmodified in cells chronically treated with the phorbol ester (protein kinase C depletion). Thus, although exogenous activation of protein kinase C results in an inhibition of insulin action, protein kinase C depletion has no influence on this process. The results suggest that, in hepatoma cells, no endogenous activation of protein kinase C may occur in response to the signal triggered by insulin.     

In vitro protein synthesis and α amylase activity in F cells from hepatopancreas ofPalaemon serratus (Crustacea; Decapoda)

In crustaceans, all the steps in the assimilation of food take place in the hepatopancreas. To facilitate the study of this organ, a method for the dissociation of cell types was developed. The hepatopancreas of the prawnPalaemon serratus was mechanically dissociated and the cells separated by Percoll density-gradient centrifugation. The E and R cells had similar densities of around 1.05 g/ml. The F cells were separated into two distinct fractions with densities of 1.075 and 1.082 g/ml. The B cells sedimented at a density of 1.12 g/ml. The ratio between the two populations of F cells was found to vary during the intermolt cycle while B cells disappeared after the molt. When the density gradient fractions were incubated with³H-leucine, incorporation was highest in the F cell fractions. Measurements of -amylase activity, indicated that the two populations of F cells may be derived from the same cell type.     

In vitro amiodarone protein binding and its interaction with warfarin

Summary The binding of amiodarone to human plasma protein and to bovine serum, albumin was studied by three different methods, ultracentrifugation, equilibrium dialysis and fluorescence spectroscopy. The fraction of amiodarone bound to plasma protein amounted to 96.3%. The changes in the binding properties of 1-anilino-naphthalene-8-sulfonate for bovine serum albumin using warfarin and amiodarone as independent inhibitors were analyzed in terms of binding site specificity. The findings indicated that amiodarone and warfarin have two different binding sites on bovine serum albumin, so a noncompetitive inhibition mechanism was indicated. On the basis of our data we cannot exclude other mechanisms of interaction besides direct displacement of one drug by another; nevertheless, metabolite interference between amiodarone and coagulation cofactors may better explain the enhancement of warfarin's pharmacological action in association with amiodarone.This work was partially funded by the CNR (National Research Council, Rome, Italy), Program on Clinical Pharmacology and Rare Diseases. The authors would like to thanks Drs E. Marzi and E. riva for their help.