Src介导骨桥蛋白诱导的血管平滑肌细胞黏附和迁移过程

为阐明酪氨酸激酶Src在整合素被骨桥蛋白(OPN)激活所触发的细胞黏附和迁移信号途径中所起的作用,应用Src特异性抑制剂PP2阻断Src,观察OPN诱导的血管平滑肌细胞(VSMC)黏附和迁移活性的改变,并利用免疫沉淀检查PP2对整合素下游信号分子黏着斑激酶(FAK)和整合素偶联激酶(ILK)磷酸化及其相互作用的影响。结果显示,PP2可明显抑制OPN诱导的VSMC黏附和伤口愈合(黏附和迁移活性分别为对照组的76.6%和33.8%);OPN可显著诱导FAK磷酸化(磷酸化水平达对照组的1.9倍),促进ILK去磷酸化,并使FAK与ILK的结合减少(降至对照组的46.4%)。10μmol/LPP2可明显抑制OPN诱导的FAK磷酸化、拮抗OPN诱导对ILK的去磷酸化作用、促进FAK与ILK之间的结合。研究结果表明,Src作为OPN-整合素-FAK信号途径中的信号分子,通过影响FAK和ILK的磷酸化以及两者之间的相互作用来调节VSMC的黏附和迁移活性。     

Ⅱ型糖尿病的红细胞膜GPA基因表达

为了研究Ⅱ型糖尿病（ＮＩＤＤＭ）患者红细胞膜血型糖蛋白Ａ（ＧＰＡ）的基因表达,采用不连续聚蔗糖（Ｐｅｒｃｏｌ）密度梯度法分离１５名正常人和２５例ＮＩＤＤＭ患者外周血的网织红细胞,提取其总ＲＮＡ,继以Ｎｏｒｔｈｅｒｎ斑点杂交观测此两组的红细胞膜ＧＰＡ基因的表达水平．采用碱性磷酸酶酶联免疫检测完成红细胞膜ＧＰ的Ｗｅｓｔｅｒｎ印迹．结果表明,ＮＩＤＤＭ患者组红细胞膜ＧＰＡ的ｍＲＮＡ含量较正常人组显著增加（分别为１１．９２±１０．５和１０．１８±１．０８积分光密度,Ｐ＜０．０１）．免疫印迹图谱显示,有６例ＮＩＤＤＭ患者的区带３ａ、３ｂ界限模糊,３例的区带３ａ、３ｂ和４明显浅染．推测ＮＩＤＤＭ患者红细胞膜ＧＰＡ减少可能引起其基因表达呈代偿性增强,而障碍或许出现在翻译环节上,这些与ＮＩＤＤＭ患者红细胞变形能力（ＲＣＤ）的明显降低甚为相符．     

一株茄病镰刀菌的代谢产物研究

一株茄病镰刀菌（Fusarium solani）固体发酵培养物经柱层析分离得到10个化合物。通过波谱分析,分别鉴定为对羟基苯甲酸甲酯（1）、水杨酸甲酯（2）、水杨酸戊酯（3）、香草乙酮（4）、草夹竹桃苷（5）、2-甲氧基-4-乙烯苯基-β-D-吡喃葡萄糖苷（6）、1-O-β-D-glucopyranosyl-(2S,3R,8E)-2-[(2R)-2-hydroxylpalmitoylamino]-8-octadecene-1,3-diol（7）、1-O-β-D-glucopyranosyl-(2S,3R,4E,8E)-2-[(2R)-2-hydroxyhexadecanoylaino]-8-octadecene -1,3-diol （8）、脑苷脂D（9）和脑苷脂C（10）。所有化合物均为首次从茄病镰刀菌中分离得到,其中化合物6首次作为天然产物报道。     

Human cell lines for biopharmaceutical manufacturing: history,status, and future perspectives

Biotherapeutic proteins represent a mainstay of treatment for a multitude of conditions, for example, autoimmune disorders, hematologic disorders, hormonal dysregulation, cancers, infectious diseases and genetic disorders. The technologies behind their production have changed substantially since biotherapeutic proteins were first approved in the 1980s. Although most biotherapeutic proteins developed to date have been produced using the mammalian Chinese hamster ovary and murine myeloma (NS0, Sp2/0) cell lines, there has been a recent shift toward the use of human cell lines. One of the most important advantages of using human cell lines for protein production is the greater likelihood that the resulting recombinant protein will bear post-translational modifications (PTMs) that are consistent with those seen on endogenous human proteins. Although other mammalian cell lines can produce PTMs similar to human cells, they also produce non-human PTMs, such as galactose-α1,3-galactose and N-glycolylneuraminic acid, which are potentially immunogenic. In addition, human cell lines are grown easily in a serum-free suspension culture, reproduce rapidly and have efficient protein production. A possible disadvantage of using human cell lines is the potential for human-specific viral contamination, although this risk can be mitigated with multiple viral inactivation or clearance steps. In addition, while human cell lines are currently widely used for biopharmaceutical research, vaccine production and production of some licensed protein therapeutics, there is a relative paucity of clinical experience with human cell lines because they have only recently begun to be used for the manufacture of proteins (compared with other types of cell lines). With additional research investment, human cell lines may be further optimized for routine commercial production of a broader range of biotherapeutic proteins.     

3种芽孢杆菌菌剂对望天树苗木生长和光合特性的影响

为加快望天树(Parashorea chinensis)幼苗的生长,采用随机区组试验设计,1.5 a生望天树实生苗分别接种胶质芽孢杆菌(Bacillusmucilaginosus)、侧孢芽孢杆菌(B.laterosporus)和巨大芽孢杆菌(B.megaterium)菌剂。结果表明,3种芽孢杆菌菌剂处理对苗木的生长、光合指标均有提高,其中苗高、地径增量分别较对照提高了30.2%～57.2%和5.3%～49.7%,根系活力提高了74.5%～227.4%,净光合速率提高了15.3%～227.6%。6.0×109cfu/g的巨大芽孢杆菌处理的苗高、地径、生物量、根表面积、根平均直径、叶面积、叶绿素含量及净光合速率、实际光合效率、光化学淬灭系数均表现最好,与苗木质量指数和隶属函数法分析结果一致。因此,6.0×109 cfu/g巨大芽孢杆菌对望天树苗木生长发育的促进作用最大。     

Three-dimensional structure of manganese superoxide dismutase from Bacillus halodenitrificans,a component of the so-called "green protein"

A so-called "green protein" has been purified from a moderate halophilic eubacterium, Bacillus halodenitrificans (ATCC 49067), under anaerobic conditions. The protein, which might play an important role in denitrification, dissociates mainly into two components after exposure to air: a manganese superoxide dismutase (GP-MnSOD) and a nucleoside diphosphate kinase. As a first step in elucidating the overall structure of the green protein and the role of each component, the 2.8-A resolution crystal structure of GP-MnSOD was determined. Compared with other manganese dismutases, GP-MnSOD shows two significant characteristics. The first is that the entrance to its substrate channel has an additional basic residue-Lys38. The second is that its surface is decorated with an excess of acidic over basic residues. All these structural features may be related to GP-MnSOD's high catalytic activity and its endurance against the special cytoplasm of B. halodenitrificans. The structure of GP-MnSOD provides the basis for recognizing its possible role and assembly state in the green protein.