Effect of Synechococcus sp.PCC7942 with trans-mutated hCu,Zn-SOD-gene on antioxidation activities in mice

目的研究转人铜锌超氧化物歧化酶(hCu,Zn-SOD)突变基因聚球藻口服后的生物活性。方法给小鼠灌服转hCu,Zn-SOD突变基因聚球藻20d,然后测定小鼠谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)和超氧化物歧化酶(SOD)活力以及丙二醛(MDA)含量。结果转hCu,Zn-SOD突变基因聚球藻可明显提高小鼠血清GSH-Px活力和全血血红蛋白CAT活性,显著提高小鼠血清和肝脏中Cu,Zn-SOD和总SOD(T-SOD)的活力,显著降低小鼠血清和肝脏的MDA含量。结论转hCu,Zn-SOD突变基因聚球藻有较强的抗氧化作用。Objective To study the biological activities of Synechococcus sp.PCC7942 with trans-mutated hCu,Zn-SOD-gene.Methods Synechococcus sp.PCC7942 with trans-mutated hCu,Zn-SOD were administered orally for 20d to mice,then the activities of glutathione peroxidase(GSH-Px),catalase(CAT),superoxide dismutase(SOD) and the content of malondialdehyde(MDA) were determined.Results The activities of GSHPx in serum and the activities of CAT in blood increased obviously;the activity of SOD in serum and liver increased markedly;the content of MDA in serum and liver decreased obviously.Conclusion Synechococcus sp.PCC7942 with trans-mutated hCu,Zn-SOD-gene had obvious antioxidant effect in vivo.福建省自然科学基金资助项目(C0510004

A study on antioxidation of synechococcus sp. PCC 7942 withTrans-thymosin α1-gene in Mice

本试验室已在蓝藻聚球藻中高效表达了人源胸腺素α1(thymosinα1,Tα1)基因,为研究转Tα1基因聚球藻口服后的生物活性,本研究给小鼠灌服转Tα1基因聚球藻14d,研究其对小鼠谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(Cat)和超氧化物歧化酶(SOD)活力以及丙二醛(MDA)含量的影响,结果表明:转胸腺素α1基因聚球藻可显著提高小鼠心、肝与肾中GSH-Px活力(P<0.01);明显提高心脏Cat活性(P<0.01);显著降低肝脏中MDA的含量(P<0.01);但对SOD活力无明显作用。提示转胸腺素α1基因聚球藻较强的抗氧化作用。Human thymosin α1 gene was expressed effectively in Synechococcus sp. PCC 7942 and antioxidant effect of Synechococcus sp. PCC 7942 with trans-thymosin α1-gene in mice were investigated. Synechococcus sp. PCC 7942 with trans-thymosin al-gene were administrated orally 14d,Laters the results showed that the activities of glutathione peroxidase (GSH-Px) in heart、liver and kidney were increased significantly (P<0. 01);the activity of catalase (Cat) in heart was increased markedly;the content of malondialdehyde (MDA) in liver was decreased obviously (P<0.01). But no significant change in the activity of super-oxide dismutase (SOD) was observed. It indicated that Synechococcus sp. PCC 7942 with trans-thymosin α1-gene had obvious antioxidation in vivo.国家海洋863课题(项目编号:819-04-03);福建省自然科学基金项目(项目编号:c0010002

Expression of recombinant hARRG cDNA in E.coli and purification of hARRG protein

目的　构建含重组人类抗砷相关基因 (humanarsenicresistencerelatedgene ,hARRG)的表达载体 ,诱导其在转化菌表达 ,分离纯化表达蛋白 ,研究该蛋白质的理化性质、抗砷功能和免疫活性 ,深入研究人类对砷化物的抵抗作用。方法　将hARRGcDNA开放阅读框亚克隆到原核表达载体Pet11C中 ,用异丙基 - β -D -硫代半乳糖苷 (IPTG)诱导表达蛋白质 ,利用阴离子交换柱Sepharose纯化蛋白质 ,SDS -PAGE胶电泳观察结果。 结果　将hARRGcDNA成功亚克隆到原核表达载体Pet11C中 ,并成功在大肠杆菌中表达 ,表达的hARRG蛋白占菌体蛋白的 5 %左右 ,该蛋白质被分离纯化。结论　原核表达载体Pet11C可以在大肠杆菌中表达hARRGcDNA ,可用阴离子交换柱Sepharose纯化抗砷相关蛋白质Objective To construct expression vector of the recombinant human arsenic resistance related gene (hARRG),induce its expression in DE\-3 and isolate and purify expression product,for studying the physiochemistry characteristic,function and immune activity of the protein,and further researching the arsenic resistant effects of human.Methods hARRG cDNA was subcloned into prokaryotic expression vector Pet11C.The recombinant protein expression was induced by IPTG,then,the protein was purified by anions Ion-exchange column Sepharose and examined by SDS-PAGE gel.Results hARRG cDNA was successfully subcloned into prokaryotic expression vector Pet11C and expressed in E.coli and the protein was purified by anions Ion-exchange column successfully.Conclusion Pet11C excpression vector containing hARRG cDNA wassuccessfully constructed,the cell DE\-3 transformed with expression vector capable of expression the gene and a hARRG protein could be purified by anions Ion-exchange column Sepharose.国家自然科学基金资助项目 (30 0 60 0 74

Bone Proteomic Analysis About Chinese Medicine Action on Rat Glucocorticoid-induced Model of Osteonecrsis.

目的:用蛋白质组学的研究方法进行激素性骨坏死的病理和中药作用机理的研究,有利于临床治激素性骨坏死。方法:按照常用激素性骨坏死的造模方法,建立大鼠坏死模型,并设立中药治疗组及空白对照组,经过6周处理后经骨形态学检查,确定骨坏死造模成功,处死动物取股骨和肱骨,提取骨组织蛋白质样品,双向电泳分离,得到各组骨组织总蛋白质分子解剖图谱,用图像分析软件,找到各组间差异蛋白质点,进一步行胶内酶切,M AD ITOF/M S质谱分析,得到各差异点的蛋白质肽指纹图谱,结合蛋白质生物信息库(M atrix sc ience L td database),对各蛋白质进行初步鉴定。结论:初步鉴定了3个差异蛋白,分别为阻凝蛋白重链ⅡB、磷脂谷胱甘肽过氧分酶及泛素化酶E 2(MW:17 kd),初步认为这三种蛋白质在激素性骨坏死的发病及中药治疗过程中发挥着重要调控作用。Objective:To study the mechanism about glucocorticoid-induced osteonecrosis and Chinese Medicine action on osteonecrosis in proteomics,it is beneficial to prevention and cure this disease.Methods:Rat was randomly divided into three groups:cortrol group,osteonecrosis model group and Chinese Medicine therapy group.In the present proteomic study ,we characterized the potential effects of glucocorticoids and Chinese Medicine on protein expression in rat bone.Using two-dimensional gel electrophoresis,mass spectrometry,and Matrix science Ltd database,we elementarily identified three variational proteins.Conclusion:Three proteins were identified as proteins similar to Myosin heavy chain ⅡB,Phospholipid hydroperoxide glutathione peroxidase and ubiquitin-conjugating enzyme E2-17kD.These proteins have been documented to be glucocorticoid-relted proteins.These results can provide valuable experimental evidences for the research of the molecular mechanism of osteonecrosis response to glucocorticoids and Chinese Medicine in bone.获得国家自然科学基金资助(30273631,3010244);; 广东省自然科学基金(020778);; 广东省教育厅基金(Z02011

Bone Proteomic Analysis About Chinese Medicine Action On Rat Ovariectomy Model of Osteoporosis.

目的:用蛋白质组学的方法对绝经后骨质疏松症和中药作用机理进行研究,有利于临床防治该疾病。方法:建立去卵巢大鼠骨质疏松症模型,设立中药治疗组及假手术对照组,6周后骨质疏松症模型成功,行骨形态学检查,提取骨组织蛋白质样品,双向电泳分离,得到各组骨组织总蛋白质分子解剖图谱,用图像分析软件,分析各组间差异蛋白质点,MADITOF/MS质谱分析,结合蛋白质生物信息库(Matrix science Ltd database),对各蛋白质初步鉴定。结论:鉴定了3个差异蛋白,分别为P1硫氧还蛋白过氧化酶1(Thioredoxin peroxidase 1),P2为阻凝蛋白轻链肽2(Myosin light polypeptide 2),P3为泛素化酶E2-17KD(ubiquitin-conjugating enzyme E2-17kD)。初步认为这三种蛋白质在绝经后骨质疏松症的发病及中药治疗过程中发挥着重要调控作用。Objective:To study the mechanism about osteoporosis of OVX(ovariectomy) and Chinese Medicine action on osteoporosis in proteomics,it is of benefit to prevent and cure this disease.Methods:Bilateal OVX in rats was performed as osteoporosis model.Rat was randomly divided into three groups:control group,osteoporosis model group and Chinese Medicine therapy group.The pathology of bone was examined after 6 weeks.In the present proteomic study,we characterized the protential effects of OVX and Chinese Medicine on protein expression in rat bones.Using two-dimensional gel electrophoresis,mass spectrometry,and Matrix science Ltd database,we elementarily identified three variational proteins.Conclusions:Three proteins were identified as proteins similar to thioredoxin peroxidase 1,Myosin light polypeptide 2 and ubiquitin-conjugating enzyme E2-17kD.These proteins have been demonstrated to be postmenopausal proteins.These results can provide valuable experimental evidences for the research for the molecular mechanism of osteoporosis which was response to OVX and Chinese Medicine in bone.国家自然科学基金项目(基金号:30400606); 广东省自然科学基金项目(基金号:04010036);; 广东省科技厅项目(基金号:粤科社字2004,139号);; 广州中医药大学创新基金项目(基金号:K004044

Preparation and determination of polyclonal antibody to thymosin α1

目的 :制备用于检测转胸腺素基因蓝藻表达产物的特异性抗体。方法 :用提纯的小肽 (2 8个氨基酸 )胸腺素α1与牛血清白蛋白偶联后作为抗原 ,采用皮下多点注射免疫的方法免疫大鼠。经过 3个月的免疫 ,获得抗Tα的多克隆抗体。结果 :用间接酶联免疫吸附 (ELISA)方法 ,测得抗体效价超过 40 96。蛋白印迹 (Westernblot)检测结果显示 ,该抗体能特异性地与胸腺素α1抗原产生明显免疫亲和反应。结论 :所制备的抗体具有很好的灵敏性和特异Objective:To prepare specific antibody against Thymosin α1 Methods:Thymosin α1(Tα1,28 peptide) was conjuncted with BSA as immune antigen,rats were immunized and the polycolonal antibody to Tα1 was obtained.Results:With ELISA detection,the titer of antibody was more than 1:4 096;Western blot analysis showed that the antibody can bind with Tα1 specifically.Conclusion:The prepared antibody against Tα1 has good specificity and reactivity and can be used to detect the expression products of transgenic cyanobacteria which expressed Tα1 gene.国家“863”课题资助!(No 819 0 4 0 3

A study of antioxdation and protective eFFect of genetical damage on phosphatidylcholine

磷脂酰胆碱500Mg/kg灌胃20日可使O_3环境中的小鼠超氧化物歧化酶（SOd）活性显著增强，血浆丙二醛（MdA）含量及外周血正染红细胞（nCE）微核率明显下降。结果提示：磷脂酰胆碱具有抗氧化作用及拮抗自由基造成遗传的损伤。进一步证实了自由基在微核形成中具有重要意义。Phosphatidylcholing(ig,500 mg/kg×20d)could elevate signiFicantly the activity of SOD and reduced remarkably the concentration of MDA and the Frequcency of NEC micrcnuleci in mice in ozone,The results indicated that phosphatidylcholine possessed ability of antioxidation and supressive eFFect on the mutagenic activity of the Free radical.This study conFirmed that the Free radical production plays an important rolein the micronucleus Fromation

Advances of research of Cyanobacterium genome

本文对近年来蓝藻基因组研究进展进行了综述.介绍了聚胞藻PCC6803基因组研究方法和成果,包括最佳分析软件的选择,所获得的基因组基本信息,以及后基因组研究的部分成果.蓝藻基因组间差异很大,文中介绍了其它蓝藻基因组的基本信息和在各个藻种里的重要发现.文章最后探讨了蓝藻基因组的研究意义和前景.The recent research advances of Cyanobacteria genome were reviewed in this paper. As the representative of genome of Cyanobacteria, the research　methods and results of genome of Synechocystis sp.strain PCC6803 wereintroduced, including the selection about most suitable analyzing soft, the genome message and some results of its post-genome research. For the much difference among cyanobacteria, the basic message and significant discovery of others Cyanobacteriagenome were described. Furthermore, the research meaning and future develoment of the Cyanobacteria genome were discussed

利用简单模板制备多孔二氧化硅

以简单有机模板为致孔剂、正硅酸乙酯(TEOS)为硅源制备了多孔SiO2,考察了模板剂加入量对样品比表面和孔容的影响,测定了样品的比表面和孔结构,提出了简单有机模板对多孔SiO2的致孔机理,并与以碱性硅溶胶为硅源对比,验证了该机理.结果表明,简单有机模板添加量为模板剂/TOES=0.3mg/mL时,样品比表面积可提高至650m2/g以上,孔容大于1.0mL/g,孔道为无规则结构.该多孔结构的产生很可能是因为简单有机模板和TEOS的聚合物发生分子级混合,从而在凝胶和煅烧过程中使样品中产生了丰富的孔道