活性依赖的突触抑制和突触稳态的分子机制

常规RNA干涉或基因敲除的功能缺失手段仅仅只是简单地移除某个基因或蛋白,而这个过程常常会掩盖磷酸化对某个特定蛋白的调节。在树突发育和突触功能活性依赖的调节过程中,突触后致密蛋白磷酸化的机制仍然是未知的领域。突触后Rap GTP酶激活蛋白SPAR与PSD95结合,可以促进树突棘的生长并加强突触。Plk2（polo-like kinase2,也称为Snk）是一种受突触活性诱导表达的蛋白激酶,它可以磷酸化SPAR,磷酸化的SPAR通过泛素化．蛋白酶体途径降解,从而导致树突棘和突触的减少。Plk2的诱导表达和随后SPAR的降解是长时间神经活性增强过程中突触强度的稳态抑制（突触剥落）所必需的。有趣的是,SPAR需要被另外一种激酶cDK5磷酸化后才能被Plk2所降解。这种机制通过CDK5对一部分突触进行标记,为由Plk2-SPAR通路抑制或去除这些突触提供了可能的途径,但其分子机制在神经退行性疾病突触丢失中的作用仍需进一步探讨。     

苹果渣基质柠檬酸高产菌株选育

分别以固态苹果渣、苹果渣固态酶解物、苹果渣酶解液和10%葡萄糖溶液为发酵基质研究了17株野生黑曲霉的柠檬酸产生能力,并对获得的4株柠檬酸高产菌进行了诱变育种。结果表明:17株黑曲霉在4种发酵基质中均能良好生长并发酵产生柠檬酸,不同菌株在同一发酵基质中产酸能力间存在差异。FG17、FG23、FG26、FG30等4株菌苹果渣基质柠檬酸产生能力较高,且在4种不同发酵基质中产酸性能稳定。4株菌分别经紫外线和60Co-γ射线诱变后得到的正向突变株柠檬酸产率均显著提高,突变株中FG26-15-4(UV)发酵苹果渣后柠檬酸产率最高,达11.32%,FG23-13-3(γ)发酵苹果渣酶解液后柠檬酸产率最高,达2.73 mg/mL,均高于现有研究报道,可作为不同类型苹果渣基质柠檬酸发酵用菌种。     

Co-expression with RadA and the characterization of stRad55B, a RadA paralog from the hyperthermophilic crenarchaea Sulfolobus tokodaii

ST0838 (designed stRad55B) is one of the four RadA paralogs (or Rad55 homologues) in the genome of the hyperthermophilic crenarchaeon Sulfolobus tokodaii. The gene is induced by UV irradiation, suggesting that it is involved in DNA recombinational repair in this organism. However, this protein could not be expressed normally in vitro. In this study, thermostable and soluble stRad55B was obtained by co-expression with S. tokodaii RadA (stRadA) in E. coli, and the enzymatic properties were examined. It was found that stRad55B bound ssDNA preferentially and had a very weak ATPase activity that was not stimulated by DNA. The recombinant protein inhibited the strand exchange activity promoted by stRadA, indicating that stRad55B might be an inhibitor to the homologous recombination in this archaeon. The results will be helpful for further functional and interaction analysis of RadA paralogs and for the understanding of the mechanism of recombinational repair in archaea.     

脆性组氨酸三联体与复制蛋白A相互作用关键氨基酸残基的鉴定

目的：研究脆性组氨酸三联体（Fhit）对ATR/CHK1通路的影响,在确定Fhit与复制蛋白A（RPA）存在相互作用的基础上鉴定Fhit与RPA相互作用的关键氨基酸残基,为进一步研究Fhit特异的信号通路奠定基础。方法：构建一系列Fhit缺失突变体基因Fhit1～Fhit11及6种Fhit点突变体基因,将这些基因插入含GST基因的原核表达载体中,在大肠杆菌中表达并纯化GST-Fhit1～GST-Fhit11融合蛋白、突变体GST-FhitSIYEEL、GST-FhitIY、GST-FhitEL、GST-FhitF、GST-FhitA,以及GST-FhitD融合蛋白,用GST沉降技术研究Fhit与RPA相互作用的关键氨基酸残基。结果：Fhit蛋白第112～117（SIYEEL）残基可能是Fhit与RPA相互作用的关键区域,而第114（Y）残基可能是Fhit与RPA相互作用的关键氨基酸残基。结论：确定了Fhit与RPA相互作用的关键氨基酸残基,为阐明Fhit在维持基因组完整性方面的机理提供了线索。     

Experimental study on the treatment of intracerebral glioma xenograft with human cytokine-induced killer cells

Objective. To investigate the phenotype changes and proliferation activities of cytokine-induced killer cells (CIKs) and lymphokine-activated killer cells (LAKs) from healthy donor, and the cytotoxicities of CIKs and LAKs to human in vitro glioma cell lines U251 and U87. Therapy of CIK intratumoral injection was evaluated in nude mouse models. Methods. CIK cells were induced from peripheral blood mononuclear cells (PBMC) of healthy donors with multiple cytokines. Phenotype analysis of CIKs and LAKs was performed with flow cytometer (FCM). The specific cytotoxicities of CIKs and LAKs against cell line U251 and U87 were determined by LDH method. After intracerebral injection of CIKs, the distribution of CIKs and the inflammatory reaction of their surrounding brain tissue were observed through continuous pathological sections. In vivo anti-tumor activity of CIKs was evaluated in athymic nude mice with intracerebral xenotransplanted U251 glioma by MRI. Results. Amount of CIKs was increased (49.83+/-2.04) times and double positive cells, CD3(+)/CD56(+) cells, were increased from (3.36+/-1.85%) to (44.07+/-14.14%) with elevated absolute amount over 1000 times after 2 week culture. In vitro experiments demonstrated that compared with LAK, CIKs possessed more obvious cytotoxic activity to U251 and U87. In vivo experiments showed that there was no severe inflammatory reaction in brain tissue. CIKs can markedly inhibit intracranial xenotransplanted glioma growth by intracranial injection (P<0.01). Conclusion. CIKs are a kind of highly effective immune cells which have a strong suppressive effect on growth for in vitro and in vivo glioma. Local injection of CIKs does not produce severe damage to normal brain tissue and is likely to be used in clinical adoptive immunotherapy of intracerebral glioma.     

湖丹皮脂溶性化学成分研究

利用柱层析方法,从湖丹皮70％乙醇提取物的氯仿萃取部位分离得到5个化合物。运用理化及波谱方法,它们分别鉴定为丹皮酚（1）、3,6-二羟基-4-甲基苯乙酮（2）、去甲基丹皮酚（3）、3,3’-二甲氧基鞣花酸（4）和胡萝卜苷（5）。其中化合物4、5为首次从该种植物中分离得到。HPLC研究表明,湖丹皮中的去甲基丹皮酚含量高于国内其它产地。     

长臀姬蜂属一新种(膜翅目,姬蜂科)

报道分布于中国的长臀姬蜂属Coleocentrus Gravenhorst,1829种类,其中含1新种:褐长臀姬蜂Coleocentrus fulvus sp.nov..描述了新种的形态特征,并与近似种做了比较,编制了中国已知种检索表.标本保存在国家林业局森林病虫害防治总站.     

作为肾素-血管紧张素系统调节器和SARS病毒受体的人血管紧张素转换酶2

人血管紧张素转换酶2(ACE2)是目前已知的惟一的人血管紧张素转换酶(ACE)的同源物,是一种新型的金属羧肽酶,很多特性与ACE截然不同.ACE2在肾素-血管紧张素系统(RAS)中具有独特的作用,调节心脏功能和机体血压.最近ACE2被鉴定为SARS病毒的功能受体.ACE2已经成为目前药物研发的新靶点.对ACE2的认识才刚刚开始,有待进-步深入研究.