Alcohol dehydrogenase 2 is a major hepatic enzyme for human retinol metabolism

The metabolism of all-trans- and 9-cis-retinol/ retinaldehyde has been investigated with focus on the activities of human, mouse and rat alcohol dehydrogenase 2 (ADH2), an intriguing enzyme with apparently different functions in human and rodents. Kinetic constants were determined with an HPLC method and a structural approach was implemented by in silico substrate dockings. For human ADH2, the determined K_m values ranged from 0.05 to 0.3 μM and k_cat values from 2.3 to 17.6 min⁻¹, while the catalytic efficiency for 9-cis-retinol showed the highest value for any substrate. In contrast, poor activities were detected for the rodent enzymes. A mouse ADH2 mutant (ADH2Pro47His) was studied that resembles the human ADH2 setup. This mutation increased the retinoid activity up to 100-fold. The K_m values of human ADH2 are the lowest among all known human retinol dehydrogenases, which clearly support a role in hepatic retinol oxidation at physiological concentrations. Received 12 October 2006; received after revision 6 December 2006; accepted 8 January 2007     

Isolation and purification of BVⅠ-2H from bee venom and analysis of its biological action

The medical use of bee venom for rheumatoid arthritis ( RA ) has a very long tradition. In this study, isolation and purification of polypeptides from bee venom were carried out on sephadex chromatography, heparin sepharose CL-6B chromatography and HPLC. Several fractions were extracted, and their effects on activation of splenocyte and THP-1 cell were studied. The inhibitory fraction was selected for further studies. Finally, BVⅠ-2H that the HPLC elution profiles was a single peak was isolated by C8 column. ESI- MS detection results showed that BVⅠ-2H was a fraction of bee venom, and the molecular weight of the major component was 644.8. BVⅠ-2H could inhibit ConA-induced splenocyte proliferation, IL-1 production and interfere with splenocyte cycle in mice. Moreover, BVⅠ-2H could inhibit PMA-induced TNFα production in THP-1 cells, which was due to its inhibitory effects on TNFα mRNA expression and protein phosphorylation of IκBα. Our studies indicated that BVⅠ-2H was one of the anti-inflammatory components of bee venom.     

Gigantecin: A novel antimitotic and cytotoxic acetogenin,with nonadjacent tetrahydrofuran rings,fromGoniothalamus giganteus (Annonaceae)

Summary Gigantecin (I), a novel tetrahydroxy-di-tetrahydrofuran fatty acid -lactone (acetogenin), was isolated from an ethanolic extract of the stem bark ofGoniothalamus giganteus Hook. f., Thomas (Annonaceae), by means of activity-directed fractionation (brine shrimp lethality test). This new compound is extremely cytotoxic to human tumor cells, inhibits crown gall tumors on potato discs, and is active in an assay designed to detect antimitotic agents (9 ASK).     

脂多糖—被动血凝试验早期诊断伤寒的价值

为评估脂多糖－被动血凝试验（ＬＰＳ－ＰＨＡ）早期诊断伤寒的应用价值,对病程不同的１０２例伤寒患者用ＬＰＳ－ＰＨＡ、肥达反应和细菌培养进行检测,２００例非伤寒发热患者作为对照。结果显示：伤寒患者病程第一周ＬＰＳ－ＰＨＡ阳性率为８０％（２４／３０）,第二周达９７７３％（４３／４４）;细菌培养阳性率２４５１％;肥达反应的伤寒与副伤寒鞭毛抗体交叉阳性率达３７２５％;对照组ＬＰＳ－ＰＨＡ有２％的假阳性。结果表明：ＬＰＳ－ＰＨＡ可早期、简便及快速诊断伤寒,有较高的敏感性和特异性。     

Cloning, construction of prokaryotic expression vector and expression ofEscherichia coli cytosine deaminase gene

Cytosine deaminase gene ofEscherichia coli strain H-30 was cloned, and its initiation codon of ‘GTG’ was mutated to ‘ATG’ by PCR. Prokaryotic recombinant expression vector pBV220-CD was constructed. Clone with high enzyme activity were selected by detecting their specific activity of cytosine deaminase. 5-FC(5-FC, 5-fluorocytosine) could induce the lethal toxicity to cells containing active CD gene. DNA sequence analysis indicated that there were 16 altered bases and 5 of them resulted in the alteration of amino acids in predicted peptide by comparing DNA sequence of the clone H-30-CD-11 with high enzyme activity with CD gene reported in Gene Bank.     

Neuropeptide Y receptors in a protozoaStylonychia mytilus

Radio ligand binding assays(RLBA) were used to study neuropeptide Y (NPY) receptors in a protozoa Stylonychia mytilus. The experimental results showed that 2-3×10+3/mL Stylonychia cells incubated in Pringsheim solution which contained 3H-NPY could specifically bind 3H-NPY and concomitantly present saturable characteristic. This suggested that Stylonychia possessed some specific binding sites for NPY. Scatchard transformations of binding assay for the NPY receptors at 25℃ are compatible with the specific activity of 42.47 fmol/10+3 cells and the binding equilibrium constant of 0.113 nmol/L. The data of 125I-NPY binding assay to the membrane protein extract of Stylonychia indicated that there was a significant difference between the amount of total bound and nonspecific bound of 125I-NPY. This result indicated that NPY receptors were probably localized mainly on the cell membrane.     

FD-TRT耐热逆转录酶的特性分析

从嗜热细菌ＦＤ３００９中克隆并分离纯化得到的耐热逆转录酶（ＦＤ－ＴＲＴ）,逆转录反应在６５℃高温下进行,可克服常温逆转录酶的许多缺点,以酵母核糖体ｒＲＮＡ为模板,以与２５５和１８Ｓ ｒＲＮＡ匹配的寡聚脱氧核苷酸为引物,探明了ＦＤ－ＴＲＴ的最适反应条件： ２５ｍｍｏｌ／Ｌ Ｔｒｉｓ－ＨＣｌ（ｐＨ ８．５,２５℃）,２５ ｍｍｏｌ／Ｌ（ＮＨ４）２ＳＯ４,２ ｍｍｏｌ／Ｌ ＭｎＣｌ２,１００ μｇ／ｍｌ明胶．５ ｕｎｉｔ ＲＮａｓｉｎ,２５０ ｍｍｏｌ／Ｌ ４种 ｄＮＴＰ, １μＣｉ３ Ｈ－ｄＣＴＰ, １２ｕｇ ＲＮＡ,２５ ｐｍｏｌ引物,并在此条件下测得 ＦＤ－ＴＲＴ和 Ｔａｑ ＤＮＡ 聚合酶逆转录活力与聚合酶活力的相对比值分别为０．０５６,０．００４５．     

用双核微核实验检测多环芳烃的致突变性

探讨了大气中常见的5种多环芳烃的致突变性。采用人外周血淋巴细胞胞质分裂阻滞微核实验检测不同浓度5种多环芳烃对人血淋巴细胞的微核诱导作用。指出5种多环芳烃各染毒剂量组微核率与阴性对照组相比有显著性差异,并且有明显的剂量反应关系,说明他们均可引起人外周血双核淋巴的微核率增加。除苯并(a)芘(BaP)外其余的几种多环芳烃也具有一定的潜在危害,应引起高度重视。     

电子受体对同步脱氮除磷的影响

在序批式反应器(SBR)系统内,利用活性污泥法,考察厌氧、缺氧、好氧组合工艺与厌氧、缺氧工艺中,硝酸盐和亚硝酸盐的存在、不同电子受体出现的顺序对同步脱氮除磷工艺的除磷影响,并讨论了在实现缺氧聚磷条件下,厌氧缺氧好氧工艺碳源的需求量。试验结果表明:反硝化聚磷是一种稳定的代谢行为,聚磷菌可以利用硝酸盐强化除磷;对于实现缺氧聚磷,硝酸盐的作用远远优于亚硝酸盐;亚硝酸盐的存在(<16.1mg.L-1)则对聚磷无明显影响;聚磷菌利用电子受体是以其存在的顺序而依次发生的,且这种利用能力不受电子受体转换的影响;在硝酸盐替代氧为电子受体的同步脱氮除磷工艺中,碳源需求将比传统工艺减少30%以上。