血啉甲醚在动脉组织的吸收分布特性

本研究观察血啉甲醚（ＨＭＭＥ）在血管平滑肌细胞（ＳＣＭ）、血管内皮细胞（ＥＣ）和动脉组织中的分布特点,旨在为建立防治血管成形术后再狭窄的光动力学疗法提供实验依据。将体外培养的ＥＣ和ＳＭＣ接种于２４忆培养板中,分组,培养液中加入ＨＭＭＥ,浓度为２０、４０、６０、８０、１００μｇ／ｍｌ,孵育１５、３０、６０、１２０、１８０、２４０ｍｉｎ后,采用荧光摄象分析法测定细胞内ＨＭＭＥ含量。以ＨＭＭＥ的孵育浓度     

两个水稻GDP-甘露糖-3’，5＇-异构酶基因特征及表达模式的研究

GDP-甘露糖-3’,5’-异构酶（GME）可以催化GDP-甘露糖转化为左旋GDP-半乳糖,该反应对于高等植物体内抗坏血酸的合成是非常重要的．但目前在分子水平上还没有对GME基因进行研究的报道．通过逆转录PCR（RT-RCR）技术从水稻成熟叶片中克隆到两个GME基因的cDNA序列,并与其他植物物种中的GMEs进行比对,结果显示,GME基因在所有植物物种中高度保守,尽管进化树分析表明单子叶植物GMEs和双子叶植物GMEs在进化上相互独立．同时,分析这两个水稻GME基因的剪切模式揭示了二者也存在高度相似性．采用半定量RT-PCR技术对两个GME基因在不同组织和不同胁迫条件下的表达模式进行研究表明,OsGME1基因在冷胁迫条件下表达水平上调,这和先前水稻冷胁迫蛋白质组学研究的结果是一致的．而OsGME2和OsGME1基因在用赤霉素处理条件下表达水平均下调,暗示赤霉素可能通过调节GME基因的表达来调控植物体内的抗坏血酸合成．     

The adipocyte as an important target cell for Trypanosoma cruzi infection

Adipose tissue plays an active role in normal metabolic homeostasis as well as in the development of human disease. Beyond its obvious role as a depot for triglycerides, adipose tissue controls energy expenditure through secretion of several factors. Little attention has been given to the role of adipocytes in the pathogenesis of Chagas disease and the associated metabolic alterations. Our previous studies have indicated that hyperglycemia significantly increases parasitemia and mortality in mice infected with Trypanosoma cruzi. We determined the consequences of adipocyte infection in vitro and in vivo. Cultured 3T3-L1 adipocytes can be infected with high efficiency. Electron micrographs of infected cells revealed a large number of intracellular parasites that cluster around lipid droplets. Furthermore, infected adipocytes exhibited changes in expression levels of a number of different adipocyte-specific or adipocyte-enriched proteins. The adipocyte is therefore an important target cell during acute Chagas disease. Infection of adipocytes by T. cruzi profoundly influences the pattern of adipokines. During chronic infection, adipocytes may represent an important long-term reservoir for parasites from which relapse of infection can occur. We have demonstrated that acute infection has a unique metabolic profile with a high degree of local inflammation in adipose tissue, hypoadiponectinemia, hypoglycemia, and hypoinsulinemia but with relatively normal glucose disposal during an oral glucose tolerance test.     

杨树胞外蛋白的提取分离及2-D电泳体系的建立

该研究以美洲黑杨杂种优良无性系NL895杨(Populus deltoides×Populus euramericana cv.)组培苗叶片和茎段为研究对象,对NL895杨叶片细胞壁蛋白(cell wall proteins,CWPs)和茎段质外体蛋白(apoplastic proteins,APPs)的提取、分离和双向电泳等技术进行了系统研究。结果表明:10g以上叶片、超声波破碎10min、CaCl2法提取的细胞壁蛋白效果较好,经G-6-PDH酶活性检测,提取的细胞壁蛋白胞质污染率较低;真空渗透法提取的茎段质外体蛋白胞质污染率较前者高,但在允许范围之内。TCA/丙酮沉淀法纯化提取的细胞壁蛋白、pH 4~7的24cm胶条、上样量为500μg的双向电泳体系,其蛋白电泳图谱中的斑点多而清晰,斑点数达550多个,是叶片细胞壁蛋白电泳分析较适合的体系;pH 3~10胶条对茎段质外体蛋白的电泳分离效果较好。该研究初步建立了杨树胞外蛋白的提取、分离及2-DE电泳体系,为木本植物胞外蛋白的研究奠定了基础。     

Biological effects of low-energy C ion implantation on sesame (Sesamum indicum L.)

Field cultivation experiments on white sesame (Sesamum indicum L.)seeds implanted with low-energy C ion showed that different dosages of C ion implantation produce different biological effects.Sesame plants in 6 different dosage groups with C ion density respectively at 1×1011,1×1012,1×1015,5×1015,1×1016,5×1016 ion/cm2 were superior to the control group in plant height,leaf number,stalk diameter and leaf size.Further,sesame plants in these groups flower and seed earlier than those in the control group,and single plant yield also increased.Of all the groups,the 5×1015 ion/cm2 dosage group yielded the best effect,whereas the 1×1017/cm2 dosage group showed an evident inhibitory effect of ion implantation on the germination and growth of the sesame seeds.     

Inhibitors of dihydrofolate reductase as antitumor agents: design,synthesis and biological evaluation of a series of novel nonclassical 6-substituted pyrido[3,2-d]pyrimidines with a three- to five-carbon bridge

Bridge homologation of the previously reported nonclassical two-carbon-bridged antifolate, 2,4-diamino-6-phenethylpyrido[3,2-d]pyrimidine (wm-5a), afforded the three-, four- and five-carbon-bridged antifolate analogues 3.1–3.5, 4.1–4.2 and 5.1–5.5. The target compounds, with substituents at various positions on the carbon bridges, were efficiently synthesized by aldol condensation or Wittig reaction and followed by reduction. Elongation of the two-carbon bridge to three-, four- or five-carbon bridges, and also saturation of the carbon bridges, provided compounds with good inhibitory activity against recombinant human DHFR (rhDHFR). Analogue 3.5, which has a three-carbon bridge, inhibited the proliferation of HL-60 and HCT116 cells to a greater extent than the other analogues. Compound 3.5 was also the most potent inhibitor of rhDHFR (IC₅₀?=?0.06?μM), and was approximately 38-fold more potent than the two-carbon-bridged lead compound. Docking studies revealed that both the length and flexibility of the saturated carbon bridge in 3.5 were important for high potency. Flow cytometry studies indicated that compound 3.5 arrested HL-60 cells in the S-phase and induced apoptosis. Western blot analysis of HL-60 cells treated with 3.5 showed a dose-dependent upregulation of DHFR protein levels.