细叶黄芪叶肉原生质体发育早期细胞壁再生的研究

采用透射电镜术、电镜多糖细胞化学染色、细胞壁荧光染色以及香豆素抑制细胞壁再生等方法,对细叶黄芪（Ａｓｔｒａｇａｌｕｓｍ ｅｌｉｌｏｔｏｉｄｅｓ ｖａｒ．ｔｅｎｕｉｓ）叶肉原生质体细胞壁的再生及其化学特点进行了研究。结果表明,离体培养２４ 小时的原生质体表面产生一些突起小泡,有时可见少量纤维组分的形成。培养３ 天时这种纤维组分明显增多。至５ 天时可清楚看到再生壁是由纤维和颗粒构成。六亚甲四胺银染色证明它们都是由多糖组分组成的。另外,培养３６ 小时的原生质体有相互粘连的现象。电镜观察、荧光染色及香豆素处理的研究表明粘连与再生壁的形成有关。根据上述观察结果,对原生质体再生壁的结构及其化学性质等问题进行了讨论     

Structure-based discovery of a novel inhibitor targeting the β-catenin/Tcf4 interaction

Overactivation or overexpression of β-catenin in the Wnt (wingless) signaling pathway plays an important role in tumorigenesis. Interaction of β-catenin with T-cell factor (Tcf) DNA binding proteins is a key step in the activation of the proliferative genes in response to upstream signals of this Wnt/β-catenin pathway. Recently, we identified a new small molecule inhibitor, named BC21 (C(32)H(36)Cl(2)Cu(2)N(2)O(2)), which effectively inhibits the binding of β-catenin with Tcf4-derived peptide and suppresses β-catenin/Tcf4 driven reporter gene activity. This inhibitor decreases the viability of β-catenin overexpressing HCT116 colon cancer cells that harbor the β-catenin mutation, and more significantly, it inhibits the clonogenic activity of these cells. Down-regulation of c-Myc and cyclin D1 expression, the two important effectors of the Wnt/β-catenin signaling, is confirmed by treating HCT116 cells with BC21. This compound represents a new and modifiable potential anticancer candidate that targets β-catenin/Tcf-4 interaction.     

高滴度单克隆抗体杂交瘤细胞系获得途径的探讨

为探索获得能分泌较高价单克隆抗体杂交瘤细胞系的途径,比较了用不同免疫方法得到的小鼠脾细胞制备杂交瘤,对其产生有效瘤和高价抗体分泌瘤的影响。用不使小鼠致病的病毒(如NDV)或巳灭活的病毒作为抗原时,以较高浓度加佐剂、长间隔(30天)、多次免疫效果较好。用能使小鼠感染的病毒作为免疫原时,以活病毒的亚致死剂量感染小鼠,取其脾细胞制备杂交瘤,效果最理想。有效杂交瘤产率高,分泌高效价抗体的杂交瘤细胞系也多。探讨了从免疫方法着手,定向研制高价单克隆抗体杂交瘤细胞系的可行性。     

抗菌肽的研究进展

抗菌肽又称抗微生物肽(antimicrobial peptide)或肽抗生素(peptide antibiotics),在动植物体内分布广泛,是天然免疫防御系统的一部分。抗菌肽不仅有广谱抗细菌能力,而且对真菌、病毒及癌细胞也有作用。对抗菌肽作用机理的研究是近来的热点之一,本文综述了此方面近来的进展,并对微生物针对抗菌肽的耐药性进行了讨论。     

毕赤酵母表达体系中重组蛋白的分离纯化

随着基因重组技术的快速发展,基因工程产品的利用越来越广泛,但其分离纯化的成本约占总成本的60%～70%.因此,探索一些简单有效的分离纯化方法尤为必要.简单介绍了目前较为流行的毕赤酵母表达体系,着重概述了重组蛋白分离纯化技术方法的应用情况.     

亚硝基胍诱变选育林肯霉素高产菌株

以林肯链霉菌９４７－８（Ｓｔｒｅｐｔｏｍｙｃｅｓｌｉｎｃｏｌｎｅｎｓｉｓ９４７－８）为出发菌株（产林肯霉素９４０γ／ｍｌ）。采用孢子热处理方法处理出发菌株孢子,得到变异株９４７－８ｓ,产林肯霉素１０８０γ／ｍｌ。对９４７－８ｓ菌株进行ＮＴＧ诱变处理,得变异株９４７－８ｘ,产林肯霉素为１２１８γ／ｍｌ,且生产能力稳定。     

1.42A crystal structure of mini-IGF-1(2): an analysis of the disulfide isomerization property and receptor binding property of IGF-1 based on the three-dimensional structure

Insulin and insulin-like growth factor 1 (IGF-1) share a homologous sequence, a similar three-dimensional structure and weakly overlapping biological activity, but IGF-1 folds into two thermodynamically stable disulfide isomers, while insulin folds into one unique stable tertiary structure. This is a very interesting phenomenon in which one amino acid sequence encodes two three-dimensional structures, and its molecular mechanism has remained unclear for a long time. In this study, the crystal structure of mini-IGF-1(2), a disulfide isomer of an artificial analog of IGF-1, was solved by the SAD/SIRAS method using our in-house X-ray source. Evidence was found in the structure showing that the intra-A-chain/domain disulfide bond of some molecules was broken; thus, it was proposed that disulfide isomerization begins with the breakdown of this disulfide bond. Furthermore, based on the structural comparison of IGF-1 and insulin, a new assumption was made that in insulin the several hydrogen bonds formed between the N-terminal region of the B-chain and the intra-A-chain disulfide region of the A-chain are the main reason for the stability of the intra-A-chain disulfide bond and for the prevention of disulfide isomerization, while Phe B1 and His B5 are very important for the formation of these hydrogen bonds. Moreover, the receptor binding property of IGF-1 was analyzed in detail based on the structural comparison of mini-IGF-1(2), native IGF-1, and small mini-IGF-1.     

Demographic and clinical variables affecting mid‐ to late‐life trajectories of plasma ceramide and dihydroceramide species

It has been increasingly recognized at the basic science level that perturbations in ceramide metabolism are associated with the development and progression of many age‐related diseases. However, the translation of this work to the clinic has lagged behind. Understanding the factors longitudinally associated with plasma ceramides and dihydroceramides (DHCer) at the population level and how these lipid levels change with age, and by sex, is important for the clinical development of future therapeutics and biomarkers focused on ceramide metabolism. We, therefore, examined factors cross‐sectionally and longitudinally associated with plasma concentrations of ceramides and DHCer among Baltimore Longitudinal Study of Aging participants (n = 992; 3960 total samples), aged 55 years and older, with plasma at a mean of 4.1 visits (range 2–6). Quantitative analyses were performed on a high‐performance liquid chromatography‐coupled electrospray ionization tandem mass spectrometer. Linear mixed models were used to assess the relationships between plasma ceramide and DHCer species and demographics, diseases, medications, and lifestyle factors. Women had higher plasma concentrations of most ceramide and DHCer species and showed steeper trajectories of age‐related increases compared to men. Ceramides and DHCer were more associated with waist–hip ratio than body mass index. Plasma cholesterol and triglycerides, prediabetes, and diabetes were associated with ceramides and DHCer, but the relationship showed specificity to the acyl chain length and saturation. These results demonstrate the importance of examining the individual species of ceramides and DHCer, and of establishing whether intra‐individual age‐ and sex‐specific changes occur in synchrony to disease onset and progression.