人α1/158Ⅴ型(α1c型)干扰素基因在大肠杆菌中的表达及其产物的初步纯化

采用DNA聚合酶链反应(PCR)技术,将本实验室从中国人胎肝细胞染色体DNA中发现和分离的IFN-α1/158V基因的原始克隆,改造成适于进行非融合蛋白原核表达的结构形式,并在大肠杆菌中获得高效表达。测得重组IFN-α1/158V的抗病毒活性为1.9×10~7单位/升菌液。随后又采用以单克隆抗体亲和层析为主的纯化流程对表达产物进行初步纯化,获得了在SDS-聚丙烯酰胺凝胶电泳上呈现单一条带的纯化产物。     

越冬期间香蕉果穗套袋的防寒效果及其对产量的影响

冬季香蕉果穗用兰色聚乙烯薄膜套袋之后,可提高袋内温度1～5℃,果实低温伤害卑只有1％左右,而未经套袋的达23％～35％,果指长度和径围增加率分别达12％和9％左右,果实产量比对照增加8％～16％。试验结果表明,兰色聚乙烯薄膜套袋是越冬期间香蕉防寒保果和增产的一项有效措施。     

Further investigation on nuclear transplantation in different orders of teleost: the combination of the nucleus of Tilapia (Oreochromis nilotica) and the cytoplasm of Loach (Paramisgurnus dabryanus).

The nucleus of a blastula cell from Tilapia (Oreochromis nilotica, family Cichlidae, order Perciformes) was transplanted into an enucleated egg of Loach (Paramisgurnus dabryanus, family Cobitidae, order Cypriniformes). From among 3747 nucleo-cytoplasmic hybrid (NCH) eggs two NCH larval fish (0.05%) were obtained; one died on the 6th day and the other died on the 12th day after the operation. Morphological examinations showed that both NCH larval fish had developed normally with an opened mouth except they could not take food after complete utilization of their egg yolk on the 5th day of development. The possible mechanisms for obtaining such inter-order NCH larval fish are discussed. This is the first report indicating that inter-order NCH larval fish can be obtained in spite of their evolutionary divergence.     

Nascent high density lipoproteins from liver perfusates of orotic acid-fed rats

Uniformly fatty livers from orotic acid-fed rats secreted almost no very low density lipoproteins (VLDL) but normal amounts of nascent high density lipoproteins (HDL) accumulated in perfusates. When lecithin:cholesterol acyltransferase (LCAT) was inhibited, nascent HDL were uniformly discoidal and lacked cholesteryl esters. Lipid and apoprotein compositions of nascent HDL from normal and fatty livers were similar whether LCAT was inhibited or not. Apolipoprotein B-100 was not detected in perfusates of uniformly fatty livers, but small amounts of apolipoprotein B-48 were present in HDL2 fractions. Nascent lipoproteins were not seen in Golgi compartments, but lipid-rich particles were clearly evident in endoplasmic reticulum cisternae adjacent to the cis face of the Golgi complex, suggesting that orotic acid blocks VLDL secretion by preventing translocation of nascent particles from the endoplasmic reticulum to the cis Golgi compartment. The accumulation of normal amounts of discoidal HDL in liver perfusates despite virtual absence of triglyceride-rich lipoproteins in Golgi secretory compartments, the space of Disse, and the perfusate is inconsistent with the concept that nascent HDL are exclusively a product of surface remnants cast off during lipolysis of chylomicrons and VLDL.     

大豆下胚轴线粒体产生超氧物自由基的效率

大豆下胚轴线粒体在呼吸基质存在下,显著地增加了肾上腺素氧化速率,这种氧化速率能为外源SOD抑制,表明线粒体呼吸时产生分子氧的单电子还原成O_2(?)。亚线粒体颗粒产生O_2(?)的效率略高于线粒体。大豆下胚轴线粒体吸链内O_2(?)的产生为NADH所支持并与交替途径无关。表明分子氧单电子还原的部位可能是NADH-黄素蛋白和UbQ-Cyt.B。     

Histopathological evidence for protective immunity induced by sonicated Salmonella vaccine

The highly susceptible inbred C3H/HeNMTV mice were vaccinated with fragments derived from sonicated Salmonella typhimurium and then infected with the pathogen. All of the vaccinated mice survived an otherwise rapidly fatal challenge of 10(5) organisms, i.e., greater than 10(3) x mean lethal dose (LD50). The vaccine also protected two-thirds of the mice infected with 10(6) bacteria and extended the survival time of the remainder in their fatal disease. Histopathological findings showed that, like the control mice, the vaccinated and infected mice developed abscesses with infiltration of polymorphonuclear leukocytes in the organs of the reticuloendothelial system during the early stage of the infection. However, unlike the primary lesions in the control mice, the lesions of the vaccinated mice tended to be discrete and self-limiting. They began to transform into granulomas after the first week of infection. Recovery and regeneration of tissues were evident 3 weeks after the infection.     

Different effects of vasopressin and angiotensin II on baroreflexes

Our data indicate that vasopressin facilitates baroreflex inhibition of lumbar sympathetic nerve activity by two mechanisms: it sensitizes baroreceptors locally and shifts the stimulus-response curve so that a lower carotid sinus pressure results in a certain level of reflex sympathetic inhibition; it also produces a corresponding shift when given i.v. and excluded from baroreceptors implicating a second, central mechanism for facilitation of baroreflexes. In contrast, angiotensin II attenuates baroreflex inhibition of peripheral sympathetic function and this is accounted for totally by a central action. Why these differences occur present challenging new questions for future investigation.     

Regulation of multiple forms of cyclic nucleotide phosphodiesterase from bovine hypothalamus: New factors modulating enzyme activity

Studies of bovine hypothalamic cyclic nucleotide phosphodiesterase (PDE) indicate the presence of several peaks of PDE activity, distinguishable by DEAE-cellulose column chromatography, displaying different substrate specificities, kinetic behavior, and regulatory properties. Evidence is presented that chromatographically separated forms of PDE activity are subject to control by Ca²⁺-calmodulin, cyclic nucleotides, limited proteolysis, reagents affecting sulfhydryl groups, and neurohormone “C”—one of several new cardioactive compounds isolated from hypothalamic magnocellular nuclei of animals—in a complex substrate-specific and concentration-dependent manner. Of particular interest is the finding that each of the forms of cGMP PDE, being Ca²⁺/calmodulin-dependent, possesses sensitivity to activation by cAMP, especially under conditions favoring the oxidation of thiol groups of PDE, resulting in a loss in responsiveness of the enzyme to the activation by calmodulin. This effect appears to be relatively stable but readily reversible by sulfhydryl reducing reagents, which restore both the cGMP PDE sensitivity to competitive inhibition by cAMP and the responsiveness of the enzyme to activation by calmodulin. A reinterpretation of the regulatory properties of multiple forms of PDE is proposed. Special Issue dedicated to Dr. Eugene Kreps.