Purification and characterization of pea chloroplastic phosphoriboisomerase

Pea (Pisum sativum L.) chloroplastic phosphoriboisomerase (EC 5.3.1.6) can be purified to apparent homogeneity in less than 2 days time with a 53% yield. Important steps in the purification include heat treatment and pseudoaffinity chromatography on Red H-3BN Sepharose. The purified isomerase has a subunit molecular mass of 26.4 kD. The N-terminal sequence has been determined through 34 residues. pH optima are 7.8 (ribose-5-phosphate) and 7.7 (ribulose-5-phosphate); K_m values are 0.9 millimolar (ribose-5-phosphate) and 0.6 millimolar (ribulose-5-phosphate). The enzyme is inhibited by erythrose-4-phosphate, sedoheptulosebisphosphate, glyceraldehyde-3-phosphate, and 3-phosphoglycerate at concentrations close to those found in photosynthesizing chloroplasts. Countercurrent phase partitioning experiments indicate that the pea chloroplastic phosphoriboisomerase interacts physically with phosphoribulokinase.     

Brefeldin A's effects on endosomes, lysosomes, and the TGN suggest a general mechanism for regulating organelle structure and membrane traffic.

Addition of brefeldin A (BFA) to most cells results in both the formation of extensive, uncoated membrane tubules through which Golgi components redistribute into the ER and the failure to transport molecules out of this mixed ER/Golgi system. In this study we provide evidence that suggests BFA's effects are not limited to the Golgi apparatus but are reiterated throughout the central vacuolar system. Addition of BFA to cells resulted in the tubulation of the endosomal system, the trans-Golgi network (TGN), and lysosomes. Tubule formation of these organelles was specific to BFA, shared near identical pharmacologic characteristics as Golgi tubules and resulted in targeted membrane fusion. Analogous to the mixing of the Golgi with the ER during BFA treatment, the TGN mixed with the recycling endosomal system. This mixed system remained functional with normal cycling between plasma membrane and endosomes, but traffic between endosomes and lysosomes was impaired.     

Cd、Fe及其复合污染对烟草叶片几项生理指标的影响

本文通过盆栽和大田模拟试验,研究了Cd,Fe,Cd+Fe,Fe+Cd处理下烟草红花大金元(Nicotiana tabacurm L.)叶片的几项生理指标的变化情况。试验表明:烟草叶片叶绿素a、叶绿素b、叶绿素a+b含量均随Cd处理浓度的增加而下降,随Fe处理浓度的增加而上升。总烟碱含量随Cd处理浓度的增加(0—100ppm)而下降,Cd 150ppm时略有上升,随Fe处理浓度的增加而上升。蛋白质含量随Cd处理浓度的增加而上升,随Fe处理浓度的增加而下降。烟草的3个生理指标表明,Cd抑制Fe对生理指标的作用,Fe抑制Cd对它们的作用,Cd与Fe相互拮抗。Cd处理的烟草叶片过氧化物酶活性和细胞膜透性的测定结果表明,过氧化物酶活性和细胞膜透性均随Cd处理浓度的增加而上升。     

用2DNMR鉴定倍半萜多元酯的结构

利用2DNMR结合其他波谱方法从苦皮藤种子油中鉴定了2个新倍半萜多元酯,它们分别是1α-苯甲酰氧-8β-烟酰氧-9α、11β-2乙酰氧-β-二氢沉香呋喃,1β-苯甲酰氧-2α、8β、9α、11β-4乙酰氧-β-二氢沉香呋喃,命名为苦皮藤酯2和3。     

毛枝南蛇藤根皮中南蛇藤素的分离鉴定及含量测定

从卫矛科南蛇藤属植物毛枝南蛇藤(Celastrus hookeri Prain)根皮中首次分离出一个红色立方体形结晶,经紫外、红外、高分辨质谱和~(13)C核磁共振谱分析,并与有关文献数据对照,确认是南蛇藤素。作者用薄层扫描和高效液相两种方法初步测定了毛枝南蛇藤根皮和根的木栓化表皮的南蛇藤素含量,结果表明南蛇藤素主要集中在根的木栓化表皮,含量在12%以上,而在整个根皮中的含量大约为1%。     

活性氧对巨噬细胞呼吸爆发影响及云芝多糖的保护作用

用化学发光法观察到叔丁基氢过氧化物对培养的小鼠腹腔巨噬细胞呼吸爆发有强烈的抑制作用。云芝多糖经腹腔注射后,能增强巨噬细胞呼吸爆发功能对叔丁基氢过氧化物损伤的抵抗力。云芝多糖处理的巨噬细胞谷胱甘肽过氧化物酶基础活力显著提高,在叔丁基氢过氧化物作用下,云芝多糖处理的巨噬细胞仍有较高的谷胱甘肽过氧化物酶活力。说明巨噬细胞的免疫功能与谷胱甘肽过氧化物酶活力有关,非特异性免疫多糖可提高细胞抗氧化能力,减轻活性氧损伤作用。     

巨噬细胞对氧化和丙二醛修饰的低密度脂蛋白结合与降解特性的比较研究

用Cu~(2+)(引发氧化修饰)和脂质过氧化降解产物丙二醛对低密度脂蛋白(LDL)进行修饰,分别测定了巨噬细胞系P~(300)D_1和小鼠腹腔巨噬细胞对两种被修饰LDL的结合量(包括内移量)和降解量。结果显示:LDL经氧化修饰和丙二醛修饰后被两类巨噬细胞的结合量与降解量均高于正常LDL,在修饰程度相近(琼脂糖电泳迁移率相近)时,两类巨噬细胞对氧化修饰LDL的结合量与降解量高于丙二醛修饰的LDL。竞争性抑制结果显示,丙二醛修饰的LDL和乙酰化修饰的LDL均可部分抑制巨噬细胞对氧化修饰LDL的结合与降解。     

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。     

改变乙型肝炎病毒核心抗原基因3′端序列对其在原核细胞中表达的影响

乙型肝炎病毒(HBV)的核心抗原基因(C基因)编码185个氨基酸残基,在原核细胞或痘苗病毒系统中能表达并装配成27nm大小的核心抗原(HBcAg)多聚体颗粒。已证实HBV C基因3′端编码近40个氨基酸的碱基序列,不是表达形成HBcAg颗粒所必需的。用外源基因替换这部分序列,已表达出表面带有外源基因产物的杂合颗粒,它具有很好的免疫原性,成为新型的基因工程多决定簇颗粒载体疫苗。但我们的实验中发现,用另外的外源基因替换3′端序列能显著影响HBV C基因在大肠杆菌中的表达,不同组成的外源基因其影响程度有所不同。     

氟中毒抑制大鼠泌鼠的实验研究

The effect of fluorosis on lactation, lactotroph function and ultrastructure were studied in lactating rats. The results were as follows: 1) Inhibition of lactation in lactating rats with chronic fluorosis was assessed by stunting growth of pups and decrease in the amount of milk suckled by pups in 30 min. Metoclopramide, a blocker of dopamine receptor, could improve lactation in these rats. 2) During chronic fluorosis serum PRL level was decreased, however, PRL content in pituitary was increased. Electronmicroscopic examination showed accumulation of large mature secretory granules and appearance of extremely large abnormal secretory granules in lactotroph cytoplasma. These findings indicate that hormone release of pituitary lactotrophs is obstructed in lactating rats with fluorosis, and the toxic effect of fluoride is mediated by an enhanced function of dopaminergic system in hypothalamus.