Biosynthesis of Territrems by Aspergillus terreus

Different radioactive precursors were added to 8-day potato-dextrose liquid cultures of Aspergillus terreus 23-1. Territrems were isolated from chloroform extracts of the cultures at day 14 and purified by thin-layer chromatography and high-pressure liquid chromatography. The territrem B obtained was treated with alkaline hydrogen peroxide, and 3, 4, 5-trimethoxy benzoic acid was isolated from an ethyl acetate extract of the reaction mixture and purified by thin-layer chromatography and high-pressure liquid chromatography. By comparison of the specific radioactivities of territrem B and its cleaved aromatic product (disintegrations per minute per micromole of compound), it was demonstrated that the radioactivity of territrem B was located mainly on its aromatic moiety when [U-C]shikimate, l-[methyl-C]methionine, and l-[methyl-H]methionine were precursors; however, the radioactivity of territrem B was located mainly on its nonaromatic moiety when [2-C]mevalonate was the precursor. Mevinolin, a specific inhibitor of beta-hydroxyl beta-methyl glutaryl coenzyme A reductase, was shown to inhibit production of territrems by A. terreus 23-1. When [U-C]acetate was used as a precursor, mevinolin inhibited the incorporation of radioactive carbon into territrem but mevinolin did not inhibit incorporation of radioactive carbon from [2-C]mevalonate into territrem.     

转基因抗虫杂交稻大米对SD大鼠行为与生理的影响

为探讨转基因抗虫杂交稻大米的食品安全性,以转cry1AC／sck双价基因抗虫杂交稻21S／MSB大米为材料．进行了大鼠30d喂养试验,检测转基因杂交大米对大鼠行为、表现体症、脏器系数、血常规与血生化指标值的影响．结果显示：转cry1AC/sck双价基因抗虫杂交稻21S/MSB大米各剂量组喂养大鼠,大鼠的行为、呼吸、毛色均正常,除雌性大鼠低剂量组、雄性大鼠中剂量组与高剂量组的胃体系数显著低于对照外,肝、脾、肾等重要脏器系数与对照组均无显著差异;受试大鼠除雌性鼠低剂量组的单核细胞计数与中剂量组的平均血小板体积、雄性鼠高剂量组的嗜中性粒细胞计数显著低于对照组,雄性鼠中剂量组的淋巴细胞计数显著高于对照组外,其余各组被测的21项血常规参数与对照组均无显著差异;除转基因大米中剂量组雌性大鼠的白球比例、低剂量组与高剂量组雄性大鼠的谷草转氨酶显著低于对照组外。其余11项被检测的血生化指标值与对照组均无显著差异．以上结果提示转cry1Ac／sck双价基因抗虫杂交稻21S／MSB大米对大鼠表观体症、脏器系数、血常规与血生化无明显不良影响．     

Human serum and glucocorticoid-inducible kinase-like kinase (SGKL) phosphorylates glycogen syntheses kinase 3 beta (GSK-3beta) at serine-9 through direct interaction

Serum and glucocorticoid-inducible kinase-like kinase (SGKL) has been identified as a new integrator that decodes lipid signals produced by the activation of phosphoinositide 3-kinase (PI3K). SGKL is activated via its lipid-binding domain (phox homology domain) in response to PI3K signaling. However, downstream targets of SGKL as well as the role of SGKL as a mediator in PI3K signaling in human tissues remain to be established. In this study, we identified human glycogen synthase kinase 3 beta (GSK-3beta) as a specific interacting partner with SGKL in a yeast two-hybrid screening of human brain cDNA library. The association between these two proteins is confirmed independently in human embryonic kidney (HEK293) cells by co-immunoprecipitation. Furthermore, the kinase activity of wild-type SGKL was required for the in vitro phosphorylation of a GSK-3 crosstide fusion protein at serine-21/9 as demonstrated with a Phospho-GSK-3alpha/beta (Ser21/9) specific antibody. The present results provide strong evidences that SGKL could utilize GSK-3beta as a direct downstream target by phosphorylating GSK-3beta at serine-9.     

Small molecules blocking the entry of severe acute respiratory syndrome coronavirus into host cells

Severe acute respiratory syndrome coronavirus (SARS-CoV) is the pathogen of SARS, which caused a global panic in 2003. We describe here the screening of Chinese herbal medicine-based, novel small molecules that bind avidly with the surface spike protein of SARS-CoV and thus can interfere with the entry of the virus to its host cells. We achieved this by using a two-step screening method consisting of frontal affinity chromatography-mass spectrometry coupled with a viral infection assay based on a human immunodeficiency virus (HIV)-luc/SARS pseudotyped virus. Two small molecules, tetra-O-galloyl-beta-D-glucose (TGG) and luteolin, were identified, whose anti-SARS-CoV activities were confirmed by using a wild-type SARS-CoV infection system. TGG exhibits prominent anti-SARS-CoV activity with a 50% effective concentration of 4.5 microM and a selective index of 240.0. The two-step screening method described here yielded several small molecules that can be used for developing new classes of anti-SARS-CoV drugs and is potentially useful for the high-throughput screening of drugs inhibiting the entry of HIV, hepatitis C virus, and other insidious viruses into their host cells.     

广西扶绥南山洞新发现的人类化石和蒙古野驴化石——末次冰期华南气候变化的哺乳动物化石新证据

广西扶绥县南山洞发现人类化石及哺乳动物化石。人类化石包括2枚智人牙齿化石,分别是左下第三臼齿和右下第二臼齿,其形态特征与广西其他山洞发现的智人化石相似,归入晚期智人。哺乳动物化石多为华南大熊猫———剑齿象动物群成员,时代属更新世晚期。该地点新发现的蒙古野驴化石属华南晚更新世动物群的首次发现。南山洞所有化石发现于洞内浅褐色砂质粘土中,堆积物第二层钙板层的铀系年龄为30—40ka。蒙古野驴的出现暗示中国南方大陆可能在末次冰期出现过干冷的气候环境。     

重组含有Epstein-Barr病毒潜伏膜蛋白1(LMP1)基因的杆状病毒在昆虫细胞中的表达

用杆状病毒表达系统重组病毒,在昆虫细胞中表达了完整的含有ＥＢＶ－ＬＭＰ１基因３个外显子开放读码框架的长２．３ｋｂ的ｃＤＮＡ片段。用重组病毒感染Ｓｆ９细胞,用免疫荧光染色,结果表明：４８小时表达重组蛋白,７２小时细胞较完整,免疫荧光染色强阳性,９６小时后细胞出现破碎。我们采集７２小时的组织培养上清和细胞破碎裂解液,分别采用ＳＤＳ－ＰＡＧＥ、ＨＰＬＣ分子筛法,用免疫蛋白印迹法实验证明,表达的蛋白能被抗ＬＭＰ１的单克隆抗体所识别,测定表达蛋白的分子量为６０ｋＤ。经蛋白含量扫描图分析,采用Ｓｅｐｈａｄｅｘ－７５柱初步纯化表达的ＬＭＰ１蛋白,将后者进行裸鼠体内致瘤实验,未见肿瘤生长。     

Potential resistance of tricin in rice against brown planthopper Nilaparvata lugens (Stål)

The aerial parts of resistant rice plant (IR36) was immersed in 85% ethanol at room temperature, and ethanol-soluble extracts were prepared. Different solvents including petroleum ether, ethyl acetate, absolute ethanol, and water were then used to separate the ethanol-soluble extracts and different solvent fractions were obtained. These fractions were tested for their effects on the mortality rate of the brown planthopper nymphae (BPH). The results indicated that EtOAc fractions had significant effects on the mortality of brown planthopper nymphae after the third day of treatment. EtOAc fractions of resistant rice plants (IR36) were separated using column chromatography over silica gel and 5,7,4′ -trihydroxy-3′,5′-dimethoxyflavone (tricin) was isolated. After the BPH were allowed to feed on diets containing tricin for 15 days, it was found that the weight of honeydew of the nymphae showed a decrease as tricin concentrations in the diets increased. The mortality rate of the nymphae increased with increasing tricin concentration. The corrected mortality rate was 58.21% for the 3rd instar nymphae and 31.75% for the 4th instar nymphae, respectively, at 500 μg/ml concentration. In addition, tricin (500 μg/ml) significantly inhibited oviposition behavior and feeding of the brown planthopper females in choice tests when spread on susceptible rice plant (TN1).     

Optimum induction of recombinant thymidine phosphorylase and its application

Recombinant E. coli pDEOA was constructed and lactose can be used instead of IPTG to induce the expression of thymidine phosphorylase by pDEOA. The use of lactose at concentrations higher than 0.5 mmol/L had an induction effect similar to that of IPTG but resulted in a longer initial induction time and better cell growth. The thymidine phosphorylase induced by lactose was very stable at 50°C. Intact pDEOA cells induced by lactose can be used as a source of thymidine phosphorylase. Under standard reaction conditions, several deoxynucleosides were effectively produced from thymidine.     

大果白刺雄性不育系的细胞学研究

利用石蜡切片法对大果白刺花药进行细胞学研究,探讨雄性不育系发生败育的时期和方式以及雄性败育与药壁组织间的关系.结果表明:不育系小孢子母细胞形成前期,花药各部分结构发育正常.随着绒毡层的异常解体,多种异常现象相继出现,包括小孢子母细胞液泡化,中层、药室内壁、药隔细胞液泡化,细胞畸形,药壁细胞非正常解体等.退化后整个花药萎缩干瘪,不能开裂,无花粉.因此,大果白刺雄性不育系的绒毡层生理异常并提前退化是导致雄性不育的主要原因.