甜菜坏死黄脉病毒的分离提纯及其生物化学特性的研究

本研究工作中,建立了一个有效的甜菜坏死黄脉病毒的分离提纯程序,解决了该病毒粒体易于聚集难以提纯的问题,其操作要点是,(1)通过Sepharose 2B柱层析代替超离心,有效地除去一些小分子量核酸杂质;(2)经PEG再次沉淀浓缩后,调整pH至酸牲(pH3.0),使病毒充分悬浮以减少凝聚;(3)在病毒等电点(pH4.8～4.9)条件下,进一步沉淀以纯化病毒。根据病毒提取物的OD260/OD280比值,算出核酸含量约4.5％。核酸电泳出现4条带,分子量分别为:2.25×10~(?),1.8×10~(?),1.05×10~(?),0.75×10~(?)道尔顿。病毒提取物经超速离心出现4个界面,沉淀系数分别为,200.8S,165S,125.8S,100S。说明甜菜坏死黄脉病毒可能是4组分病毒粒体。病毒粒体含一蛋白亚基,分子量约为2.05±0.05×10~4道尔顿,由16种共199个氨基酸组成。     

流行性感冒病毒重组株京生75-29 R2 T1及冷适应株31-广的基因分析

用聚丙烯酰胺凝胶电泳方法分析了流行性感冒病毒重组株京生75-29R2 T1(H3N2)及冷适应株31-广(H3N2)的RNA及多肽。重组株京生75-29R2 T1的HA及M基因系来自流行病毒亲本株/甲/北京/29/75(H3N2),而P_2、NA、NP及NS基因则来自温度敏感母株福R3(H2N2)。流行病毒株甲/穗/03/68(H3N2)在低温条件下经鸡胚尿囊腔传递24代而获得的冷适应疫苗毒株31-广(H3N2)其基因型与野毒株一致。     

北京东郊作物土壤系统中重金属的迁移、分布、积累

 本文研究了北京东郊污灌区重金属在作物—土壤中的迁移、分布、积累规律,证实本区蔬菜中汞含量比粮食作物约大3—15倍,比水果约大6—200倍。麦粒、糙米中的Cu、Hg、Cd、Pb、Ni的含量与土壤含量相关性不显著。架豆中重金属含量与土壤中重金属含量的相关性,只有Zn,Pb达显著水平。白菜土有机质含量与重金属含量相关性达显著水平,而白菜的重金属含量与土壤的重金属含量相关性却不显著。说明除了土壤中重金属的总量外,有效态含量的多少,是影响本区作物吸收积累重金属的主要因素。 本区施污泥的土壤和生长的作物Cd／Zn大部小于1%、盆栽试验证明：施用本区污泥污水对水稻生长发育的影响比施污泥灌清水的影响大些,因此,施用含重金属污泥时,最好不要超过5000斤／亩。大田和室内模拟试验证明：重金属从土壤中迁移到植物,由植物带走输出的量极少,其中以带走输出的Hg、Cd,As相对较多,带走输出的Pb、Cr相对的少些。     

Sperm-oocyte membrane fusion in the human during monospermic fertilization

Sperm-oocyte membrane fusion has been observed during monospermic fertilization of a human oocyte in vitro. Women were stimulated with both clomiphene citrate and human menopausal gonadotropin and were given human chorionic gonadotropin before a LH-surge. Twelve oocytes, collected at laparoscopy from six women who became pregnant by IVF, were allowed to mature for 7–14 hours in vitro and inseminated with preincubated sperm, fixed between 1–3 hours after insemination, and examined by transmission electron microscopy. Membrane fusion had occurred in one ovum 2 hours after insemination, and the oocyte had resumed maturation and was at anaphase II of meiosis. Cortical granules had been exocytosed, and some of their contents were visible at the surface close to the oolemma all around the oocyte. The sperm that fused with this oocyte was acrosome-reacted and had been partly incorporated into the ooplasm, while the anterior two-thirds of its head was phagocytosed by a tongue of cortical ooplasm. Membrane fusion had occurred between the oolemma and the plasma membrane overlying the postacrosomal segment of the sperm head, posterior to the equatorial vestige. Sperm chromatin had not decondensed, and serial sections revealed a midpiece attached to the basal plate and a tail located deeper in the ooplasm, all devoid of plasma membrane. Supplementary sperm penetrating the inner zona, approaching the perivitelline space, had undergone the acrosome reaction but had a persistent vestige of the equatorial segment of the acrosome with intact plasma membrane. Evidence of sperm chromatin decondensation was seen in other oocytes, 3 hours after insemination, which were at telophase II of meiosis. Eight oocytes penetrated by sperm were monospermic, while four were unfertilized. The general pattern of sperm fusion and incorporation appears to conform to that seen in most other mammals. The study also reveals that sperm have to complete the acrosome reaction before fusing with the egg.     

HBsAg合成肽段的免疫源性和抗肽血清与天然HBsAg的结合测定

我们将三种乙型肝炎表面抗原的合成肽段在联结载体或不接载体的情况下,分别制备成4种免疫原对兔子进行了免疫。大部分兔子都产生了抗肽抗体。其中抗P_(122-48)(auw)与抗P_(122-148)(adw)都能与天然HBsAg反应,而抗P_(122-148)(adw)的抗HBs活力比文献报道的都高,通过结构分析表明:含有天然蛋白上免疫显性区域的免疫原是合成疫苗的理想选者。     

百日咳杆菌去内毒素及保护性抗原的纯化

内毒素(Endotoxin,简称ET)是百日咳全菌苗(Bordetellapertussis vaccine)产生副作用的主要毒素之一,且不易除去。现有的分离方法,如蔗糖密度梯度离心法,较繁琐,成本高。本文采用Sepha-cryl S-300凝胶层折法可以简便有效的去除大部分内毒素。初步毒素试验结果表明:已达到日本生物制品规格的要求。两种保护性抗原FHA和LPF-HA也得到进一步分离纯化,为今后研制高效的百日咳组分菌苗提供了实验条件。     

Metabolism of viprostol — A synthetic vasolidator PGE2 analog

Metabolic studies were done with ¹⁴C-Viprostol (I) administered by various routes (I.V., oral and topical) to six animal species and to man. Total radioactivity and metabolic profiles were analyzed in plasma, tissues and excreta. The main metabolites were isolated and identified by capillary GC/MS.Plasma and urinary metabolic profiles were qualitatively similar across species, with two major metabolic reactions being predominant: rapid hydrolysis to the pharmacologically active free acid (II)_ and oxidation of the alpha-chain to dinor and tetranor acids (III, IV). In the monkey and man, reduction of the 9-keto group lead to PGF₂ type metabolites (VI–VIII). In the rat, omega oxidation of the beta-chain occurred as well, resulting in the formation of dicarboxylic acids (V).     

从棒头草(Polypogon fugax Nees ex Steud)幼穗培养获得体细胞胚和再生植株

棒头草幼穗在含2,4-D的MS培养基上诱导出了胚性、非胚性和中间型愈伤组织。根据形态、淀粉粒等指标可将组成这些愈伤组织的细胞分为三类。改变培养基中2,4-D的浓度,能诱导三类愈伤组织相互转变。从胚性愈伤组织中诱导形成了大量体细胞胚;体细胞胚是从单个原胚细胞直接发育而来,它们能正常萌发、再生小植株。这种再生能力现已保持了34个月。小植株移植在土壤中可以正常生长、分蘖、开花和结实。     

B7-2胚胎性癌细胞诱导分化过程中的表型转变

胚胎性癌细胞(简称EC细胞)作为一类肿瘤(畸胎瘤)的干细胞近年受到广泛的重视,从胚胎学、肿瘤学和分子生物学等许多学科领域都应用它作为实验材料,离体诱导分化研究是其中的一个方面。B 7-2 EC细胞是我们从129品系小鼠的自发睾丸畸胎瘤中分离克隆得到的一株多能EC细胞,它在同种同基因小鼠     

Design of a system for the control of low dissolved oxygen concentrations: critical oxygen concentrations for Azotobacter vinelandii and Escherichia coli

The physiological activity of microorganisms in environments with low dissolved oxygen concentrations often differs from the metabolic activity of the same cells growing under fully aerobic or anaerobic conditions. This article describes a laboratory-scale system for the control of dissolved oxygen at low levels while maintaining other parameters, such as agitator speed, gas flowrate, position of sparger outlet, and temperature at fixed values. Thus, it is possible to attribute in dilute nonviscous fermentations all physiologic changes solely to changes in dissolved oxygen. Experiments were conducted with Azotobacter vinelandii and Escherichia coli. Critical oxygen concentrations for growth (that value of oxygen allowing growth at 97% of mu max) were measured as 0.35 +/- 0.03 mg/L for A. vinelandii and 0.12 +/- 0.03 mg/L for E. coli. These values are significantly different from the commonly quoted values for critical oxygen concentrations based on respiration rates. Because of the superior dissolved oxygen control system and an improved experimental protocol preventing CO2 limitation, we believe that the values reported in this work more closely represent reality.