我国北方异色瓢虫越冬集群的调查研究:越冬场所的调查初报

在毛主席革命路线指引下,我国农林害虫的生物防治得到了迅速发展。为了在本世纪内,全面实现农业、工业、国防和科学技术现代化,在植物保护工作上,我们要积极贯彻“预防为主,综合防治”的方针,采取综合防治消灭害虫。“以虫治虫”是其中的重要措施。研究益虫越冬集群的规律,扩大益虫来源,给利用天敌提供丰富的物质基础,为此开展本题的调查研究。     

多光束中医信息治疗仪的研制

本文介绍了一种应用光子中医学技术防治缺血性疾病的多光束弱激光中医信息治疗仪。该仪器将先进的单片计算机技术,激光技术与光子中医信息疗法融为一体。文章主要介绍了多光束中医信息治疗仪的设计理念及其实现方式。     

人血清蛋白质组学方法的比较和优化

目的:对人未做处理的血清以及去除白蛋白和免疫球蛋白G(IgG)血清的蛋白质组学方法进行比较和优化。方法:应用双向电泳(2-DE)方法分离了未做处理的以及去除白蛋白和免疫球蛋白G(IgG)的血清,比较优化了高温变性、水化液成份组成及泡胀方式等影响血清2-DE分离效果的因素,并用质谱分析鉴定未做处理和已处理血清的2-DE谱图中部分差异蛋白点。结果:得到了分辨率和重复性较好的2-DE谱图,未做处理的血清、去除白蛋白及IgG血清的平均蛋白质点分别为(482±18)个和(523±29)个,质谱分析了9个差异蛋白点,鉴定为8种蛋白质,其中7种为功能蛋白质。仅出现在未做处理血清中的蛋白有4种,分别是维生素A结合蛋白、可溶性尿激酶血纤维蛋白溶酶原激活剂受体、蛋白激酶1抗原、血清白蛋白。4种蛋白仅出现在去除白蛋白和IgG的血清中,分别是NADH脱氢酶辅酶β亚基、肌动蛋白结合蛋白M1、T细胞活性受体β链、血小板生长因子C。结论:去除高丰度蛋白可增加一些低丰度蛋白质的检出,但非特异性吸附会导致部分功能蛋白质的丢失。     

7β-Methyl substituent is a structural locus associated with activity cliff for nepenthone analogues

With the purpose of identifying novel selective κ opioid receptor (KOR) antagonists as potential antidepressants from nepenthone analogues, starting from N-nor-N-cyclopropylmethyl-nepenthone (SLL-020ACP), a highly selective and potent KOR agonist, a series of 7β-methyl-nepenthone analogues was conceived, synthesized and assayed on opioid receptors based on the concept of hybridization. According to the pharmacological results, the functional reversal observed in orvinol analogues by introduction of 7β-methyl substituent could not be reproduced in nepenthone analogues. Alternatively, introduction of 7β-methyl substituent was associated with substantial loss of both subtype selectivity and potency but not efficacy for nepenthone analogues, which was not found in 7β-methyl orvinol analogues. Surprisingly, SLL-603, a 7β-methyl analogue of SLL-020ACP, was identified to be a KOR full agonist. The possible molecular mechanism for the heterogeneity in activity cliff was also investigated. In conclusion, 7β-methyl substituent was a structural locus associated with activity cliff and demonstrated as a pharmacological heterogeneity between nepenthone and orvinol analogues that warrants further investigations.     

Two fused proteins combining Stichopus japonicus arginine kinase and rabbit muscle creatine kinase

Two fused proteins of dimeric arginine kinase (AK) from sea cucumber and dimeric creatine kinase (CK) from rabbit muscle, named AK-CK and CK-AK, were obtained through the expression of fused AK and CK genes. Both AK-CK and CK-AK had about 50% AK activity and about 2-fold K _m values for arginine of native AK, as well as about 50% CK activity and about 2-fold K _m values for creatine of native CK. This indicated that both AK and CK moieties are fully active in the two fused proteins. The structures of AK, CK, AK-CK, and CK-AK were compared by collecting data of far-UV circular dichroism, intrinsic fluorescence, 1-anilinonaphthalene-8-sulfonate binding fluorescence, and size-exclusion chromatography. The results indicated that dimeric AK and CK differed in the maximum emission wavelength, the exposure extent of hydrophobic surfaces, and molecular size, though they have a close evolutionary relationship. The structure and thermodynamic stability of AK, CK, AK-CK, and CK-AK were compared by guanidine hydrochloride (GdnHCl) titration. Dimeric AK was more dependent on the cooperation of two subunits than CK according to the analysis of residual AK or CK activity with GdnHCl concentration increase. Additionally, AK and CK had different denaturation curves induced by GdnHCl, but almost the same thermodynamic stability. The two fused proteins, AK-CK and CK-AK, had similar secondary structure, tertiary structure, molecular size, structure, and thermodynamic stability, which indicated that the expression order of AK and CK genes might have little effect on the characteristics of the fused proteins and might further verify the close relationship of dimeric AK and CK. Published in Russion in Biokhimiya, 2006, Vol. 71, No. 9, pp. 1208–1214.     

低强度激光联合照射犊鼻穴和口咽部对膝骨关节炎兔细胞因子IL-1β及TNF-α的影响

目的：探讨携带针灸补泻信息的激光联合多部位照射对膝骨关节炎兔血清和关节液中细胞因子的影响。方法：40只新西兰兔随机分成对照组、模型组、穴位照射组和联合照射组。兔左侧膝关节腔内注入木瓜蛋白酶造成关节炎的病理模型,穴位照射组和联合照射组分别以低能量激光（输出功率10—20mW）连续照射犊鼻穴和携带针灸补泻信息的脉冲激光联合照射口咽部及犊鼻穴,10min／d,5d为1个疗程,连续3个疗程。采用放射免疫的方法测定血清及关节液中相关细胞因子含量的变化,常规病理学切片观察膝关节软骨组织形态学变化。结果：与对照组比较,模型组血清和关节液中IL-1B及TNF—α含量均显著升高（P〈0．01）;与模型组比较,穴位照射组和联合照射组血清和关节液中IL-1β及TNF—α含量均下降,但以联合照射组效果显著（P〈0．05）;穴位照射组和联合照射组之间的IL-1B及TNF-饯含量未表现出显著性差异（P〉0．05）。结论：激光联合照射口咽部和犊鼻穴和激光照射犊鼻穴都能减少细胞因子IL-1β及TNF-α的生成,减轻关节软骨细胞的结构损伤,起到保护关节软骨、防治骨性关节炎的作用,但联合多部位照射效果优于单独穴位照射。     

Inactivation and conformational changes of lactate dehydrogenase from porcine heart in sodium dodecyl sulfate solutions

The inactivation and conformational changes of porcine heart lactate dehydrogenase (LDH) have been studied in sodium dodecyl sulfate (SDS) solutions. Increasing SDS concentration led to a quick and concentration-dependent inhibition of the enzyme, with complete inactivation within 5 min in the presence of 1.0 mM SDS. Meanwhile, fluorescence emission and circular dichroism spectra were used to follow the conformational changes of the enzyme during this process, concurrently showing that SDS less than 1.0 mM induced only limited conformational changes to LDH. The above results are in accordance with the suggestion by Tsou (Trends Biochem. Sci. 11 (1986) 427; Science 262 (1993) 380) that the active site usually be more flexible than the enzyme molecule as a whole. Furthermore, the results of polyacrylamide gel electrophoresis (PAGE) implied that unfolding intermediates were presented in the above process. When the SDS concentration used to treat LDH was increased, the bands of native enzyme on native PAGE faded and finally almost disappeared. Meanwhile, multiple bands with lower mobility but no activity emerged behind and enhanced correspondingly. Fast protein liquid chromatography indicated that dissociation occurred during the course of denaturation. The reasons for the above phenomena have been discussed. It was suggested that SDS, binding to LDH to form different LDH-SDS complexes, conferred an array of different unfolding states over the enzyme, and in turn resulted in the formation of the multiple bands on the native PAGE.     

氨基酰化酶Ⅰ的必需半胱氨酸巯基

本文用DTNB和IAM修饰了猪肾氨基酰化酶Ⅰ的半胱氨酸巯基,用Koshland和邹承鲁作图法定量处理的结果都表明,酶分子中有二个巯基为酶的必需巯基。     

HLA-G阳性的胎盘间充质干细胞体外诱导Treg的实验研究

目的：研究HLA-G阳性的胎盘间充质干细在体外诱导Treg的产生。方法：从新生儿胎盘中分离胎盘间充质干细胞的,采用脂质体转染的方式将PEGFP-N1-HLA-G质粒转染到胎盘间充质干细胞中,将细胞分为空白对照组、PEGFP-N1组和PEGFP-N1-HLA-G组,每组设置5个复孔,并通过蛋白质免疫印迹检测HLA-G的表达,将鉴定后的细胞与健康人外周血中CD4⁺的T淋巴细胞混合培养24 h和48 h,并检测CD4⁺CD25⁺Foxp3⁺Treg占T淋巴细胞的比例。结果：PEGFP-N1-HLA-G转染后胎盘间充质干细胞可以表达HLA-G蛋白,与空白对照组和PEGFP-N1组相比有显著性差异（P<0.01）;HLA-G阳性的胎盘间充质干细胞在与CD4⁺的T淋巴细胞混合淋巴细胞培养24 h后,Treg细胞占全部T淋巴细胞的比例为（16.41±0.94）%,在培养48 h后,Treg细胞的占全部T淋巴细胞的比例为（16.46±0.59）%,与空白对照组和PEGFP-N1组相比有显著性差异（P<0.01）。结论：HLA-G基因修饰后胎盘间充质干细胞能够有效的在体外诱导CD4⁺ CD25⁺ FoxP3⁺Treg产生。     

N-乙酰半胱氨酸体外对流感病毒H1N1的抑制作用

目的探讨N-乙酰半胱氨酸（N—acetylcysteine,NAC）在体外对流感病毒H1N1的抑制作用。方法采用MTT法、鸡胚接种法和免疫荧光法,观察NAC对流感病毒HlM的抑制作用。采用血球凝集试验、神经氨酸酶活性抑制试验和透射电镜负染技术,初步探讨NAC对流感病毒HlM的抑制机制。结果NAC在MDCK细胞上的最大无毒剂量是6．25mg／mL;流感病毒H1Nl在MDCK细胞上的半数致死感染浓度（TCID-50）为1012-2.25／100μ;在三种作用途径下（治疗性给药、预防性给药和直接灭活后给药）,NAC明显抑制了流感病毒HlNl对MDCK细胞的感染,细胞存活率分别为91．88％、93．21％、94．67％,在对照组,流感病毒H1N1感染后的细胞存活率为28．32％,两者相比差异有统计学意义（P〈0．05）;免疫荧光结果显示,与病毒对照组形成的强特异性荧光相比,三种作用途径感染MDCK细胞后的特异性荧光明显减弱;鸡胚培养法的结果显示,NAC明显抑制了流感病毒H1N1在鸡胚内的增殖,实验组血凝效价低于1：2,对照组血凝效价为1：1024;神经氨酸酶活性抑制试验和透射电镜的结果显示,NAC能够明显抑制流感病毒川N1的神经氨酸酶活性,对流感病毒H1N1的病毒体结构也有明显的破坏作用。结论NAC在体外对流感病毒川N1有明显的抑制作用,其抑制机制可能与NAC对流感病毒血凝素和神经氨酸酶活性抑制及病毒体的直接破坏有关。