Cloning and expressing DBT (dibenzothiophene) monooxygenase gene (dszC) from Rhodococcus sp. DS-3 in Escherichia coli

Dibenzothiophene (DBT) monooxygenase (DszC)catalysis,the first and also the key step in the microbial DBT desulfurization,is the conversion of DBT to DBT sulfone (DBTO2).In this study,dszC of a DBT-desulfiaizing bacterium Rhodococcus sp.DS-3 was cloned by PCR.The sequence cloned was 99% homologous to Rhodococcus erythropolis IGTS8 that was reported in the Genebank.The gene dszC could be overexpressed effectively after being inserted into plasmid pET28a and transformed into E.coli BL21 strain.The expression amount of DszC was about 20% of total supernatant at low temperature.The soluble DszC in the supematant was purified by Ni2+ chelating His-Tag resin column and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to electronics purity.Only one band was detected by Western-blotting,which is for the antibody released in mouse against purified DszC in the expression product of BL21 (DE3,paC5) and Rhodococcus sp.DS-3.The activity of purified DszC was 0.36 U.DszC can utilize the organic compound such as DBT and methyl-DBT,hut not DBT derivates such as DBF,which has no sulfur or inorganic sulfur.     

A conserved domain of herpes simplex virus ICP34.5 regulates protein phosphatase complex in mammalian cells

ICP34.5, encoded by herpes simplex virus 1, is a protein phosphatase 1 (PP1) regulatory subunit that mediates dephosphorylation of the alpha subunit of translation initiation factor 2 (eIF2alpha). However, the mechanism of its action remains poorly understood. Here, we show that amino acid substitutions in the arginine-rich motif have differential effects on ICP34.5 activity. The phenotypes parallel with viral protein synthesis and cytopathic effects in virus infected cells. Besides the consensus PP1 binding motif, the Arg-motif appears to enhance the interaction between ICP34.5 and PP1. These results suggest that concerted action between the PP1 binding domain and the effector domain of ICP34.5 is crucial for eIF2alpha dephosphorylation and viral protein synthesis.     

Protective effects of leukemia inhibitory factor against oxidative stress during high glucose-induced apoptosis in podocytes

Leukemia inhibitory factor (LIF) is a pleiotropic glycoprotein belonging to the interleukin-6 family of cytokines. In kidney, LIF regulates nephrogenesis, involves in tubular regeneration, responds to pro- and anti-inflammatory stimuli, and so on. LIF also plays an essential role in protective mechanisms triggered by preconditioning-induced oxidative stress. Although LIF shows a wide range of biologic activities, effects of LIF on high glucose-induced oxidative stress in podocytes remain unclear. The aim of the study was to assess whether LIF can attenuate high glucose-induced apoptosis in podocytes. The result of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay indicated that LIF protected podocytes against high glucose-induced cytotoxicity. The flow cytometry assay showed that LIF attenuated high glucose-induced apoptosis in podocytes. Meanwhile, the result of flow cytometric assay gave the clear indication that LIF decreased high glucose-induced elevated level of reactive oxygen species (ROS). The measurement of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, superoxide dismutase (SOD), malondialdehyde (MDA), and caspase-3 activity levels showed that LIF attenuated the high glucose-induced decreased level of SOD and elevated level of NADPH oxidase, MDA and caspase-3 activity. These results may provide potential therapy for diabetic nephropathy in the future.     

中国真开氏摇蚊属一新种及一新记录种记述（双翅目：摇蚊科）

记述真开氏摇蚊属Eukiefferiella Thienemann 1新种:盖真开氏摇蚊E. celata sp. nov.和1种中国新记录种: 翘叶真开氏遥蚊E. brehmi Gouin, 1943;提出兵库真开氏摇蚊E. togaeuquinta Sasa & Okazawa, 1992为翘叶真开氏摇蚊 E. brehmi Gouin,1943的新异名。     

花生四烯酸代谢相关酶和ERK与乳腺癌转移关系

花生四烯酸代谢相关酶环加氧酶(COX)、脂氧合酶(LOX)和活化的胞外信号调节激酶（p-ERK1/2）等与乳腺癌发生发展具有密切关系.为了进一步阐明它们在乳腺癌转移中的作用及其分子机制,应用反转录PCR（RT-PCR）、免疫印迹和免疫组化等方法,分别检测了乳腺癌细胞系、乳腺癌组织、癌旁组织和转移淋巴结组织中COX-2、5-LOX、12R-LOX、cPLA2和p-ERK1/2的表达水平.结果显示,与对照组相比,在具有高转移潜能的乳腺癌LM MCF-7细胞和MDA-MB-231细胞以及转移淋巴结组织中,COX-2、5-LOX、12R-LOX、cPLA2和p-ERK1/2均有较高水平表达.进而发现,p-ERK1/2的特异性抑制剂PD98059可拮抗LM-MCF-7细胞和MDA-MB-231细胞中COX-2、5-LOX、12R-LOX和cPLA2的高表达.上述结果表明,p-ERK1/2可以通过促进花生四烯酸代谢相关酶的表达促进乳腺癌细胞发生转移.     

内蒙古高原西部荒漠区锦鸡儿属(Caragana)优势种的形态适应特征

研究内蒙古高原西部荒漠区锦鸡儿属（Caragana）4种优势植物——柠条锦鸡儿、狭叶锦鸡儿、垫状锦鸡儿和荒漠锦鸡儿的地理分布和叶形态适应特征.通过地理分布和生长发育研究发现：4种锦鸡儿均能在阿拉善荒漠区分布和正常生长,它们能够适应阿拉善荒漠区环境.叶形态结构研究表明：狭叶锦鸡儿叶片呈瓦状,在一定程度上阻碍了水分散失和气体交换,缩小受光面积;垫状锦鸡儿叶片卷成筒状,水分散失和气体交换阻力更大,受光面积更小.柠条锦鸡儿密被伏帖绢毛、狭叶锦鸡儿多被灰白色柔毛、垫状锦鸡儿密被绢毛和荒漠锦鸡儿密被直立绢毛,对光线反射能力强,使叶温降低,减少蒸腾.狭叶锦鸡儿和垫状锦鸡儿具有的长条型叶,4种荒漠区锦鸡儿都具有更小的叶面积、更大的厚度/面积比值、更低的叶片生物量和更小的比叶面积（SLA）,减小了蒸腾和受光面积,提高了荒漠区锦鸡儿的保水能力.荒漠区锦鸡儿这些叶片形态特性有利于适应干旱、高温和强辐射的荒漠区环境.叶绿素含量测定和叶色研究结果表明：4种荒漠区锦鸡儿叶绿素含量低,更低的叶绿素含量和密被灰白色柔毛或绢毛导致叶色浅、反光性能好,有利于减少对辐射的吸收,避免强辐射对植物的伤害,同时也使叶温低,减轻高温伤害和水分蒸发,以适应环境.这是荒漠区锦鸡儿适应高光强、高气温、极干旱的荒漠区环境的特性.垫状锦鸡儿和柠条锦鸡儿叶绿素a/b高也是适应强辐射的特性.比较4种荒漠区锦鸡儿的叶形态特性发现,4种荒漠区锦鸡儿植物对荒漠环境的形态适应方式是不同的.     

Synthesis, characterization and biological activity of lanthanum(III) complexes containing 2-methylene-1,10-phenanthroline units bridged by aliphatic diamines

Two novel lanthanum(III) complexes containing 2-methylene-1,10-phenanthroline units bridged by aliphatic diamines were synthesized and characterized by elemental analysis, IR, NMR, thermal analysis and conductance measurements. They have been assayed for anticancer activity in vitro against HL-60 (human leukocytoma) cells, PC-3MIE8 (human prostate carcinoma) cells, BGC-823 (human stomach carcinoma) cells, MDA-MB-435 (human galactophore carcinoma) cells, Bel-7402 (human liver carcinoma) cells, and Hela (human cervix carcinoma) cells. The results show that the two complexes exhibit good cytotoxic activities against different cell lines in general, especially more effective than cisplatin against Bel-7402, BGC-823 and MDA-MB-435 cell lines. DNA-binding studies indicate that, besides the intercalation, the complexes bind to DNA by the other interaction(s), which might be responsible for the production of more compact DNA, coinciding with more A-like feature of DNA as suggested by CD spectra.     

Peptidase inhibitors from the salivary glands of the cockroach Nauphoeta cinerea

Inhibitory activity against subtilisin, proteinase K, chymotrypsin and trypsin was detected in the salivary glands and saliva of the cockroach Nauphoeta cinerea (Blattoptera: Blaberidae). Fractionation of the salivary glands extract by affinity chromatography followed by reverse-phase HPLC yielded five subtilisin-inhibiting peptides with molecular masses ranging from 5 to 14 kDa. N-terminal sequences and subsequently full-length cDNAs of inhibitors designated NcPIa and NcPIb were obtained. The NcPIa cDNA contains 216 nucleotides and encodes a pre-peptide of 72 amino-acid residues of which 19 make up the signal peptide. The cDNA of NcPIb consists of 240 nucleotides and yields a putative secretory peptide of 80 amino-acid residues. Mature NcPIa (5906.6 Da, 53 residues) and NcPIb (6713.3 Da, 60 residues) are structurally similar (65.4% amino acid overlap) single-domain Kazal-type peptidase inhibitors. NcPIa with Arg in P1 position and typical Kazal motif VCGSD interacted stoichiometrically (1:1) with subtilisin and was slightly less active against proteinase K. NcPIb with Leu in P1 and modified Kazal motif ICGSD had similar activity on subtilisin and no on proteinase K but was active on chymotrypsin.     

铜绿假单胞菌色素代谢相关基因的研究

首次应用Mu转座重组技术研究铜绿假单胞菌色素合成与调控的机制。通过一系列的表型筛选,得到8株色素合成能力改变的突变子。经基因克隆、核苷酸测序研究,证明转座子分别插入到hmgA、ptsP、sucC、phzS、phzF1五个基因中。hmgA基因转座失活导致酪氨酸分解代谢中间产物尿黑酸的积累,后四种情况转座突变显著地影响了铜绿假单胞菌最重要的色素绿脓素（pyocyanin）的合成,其中PhzS和phzF1是绿脓素合成过程中的结构基因,ptsP基因是1个磷酸转移酶系统的重要组分,sucC基因的产物是三羧酸循环中的琥珀酰辅酶A合成酶,对后两个基因在色素合成的调控方面可能起到重要作用的报道尚属首例。