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Journal of pharmaceutical Analysis 2010年第2期22卷 97-101页

作者： Qin Xiao1,2.Hui Chen2.Jin-Ming Lin2.1. college of Ocean,Hebei Agricultural University,Qinhuangdao 066003 2. The Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology,Department of Chemistry,Tsinghua University,Beijing 100084,china. college of Ocean Hebei Agricultural University Qinhuangdao 066003 China Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology Department of Chemistry Tsinghua University Beijing 100084 China

A simple and rapid expression and purification method of recombinant firefly luciferase was developed for bacteria detection. A modified luciferase gene from North American firefly Photinus pyralis was cloned into pET2.a expression vector and the recombinant protein was produced in Escherichia coli BL2.. The recombinant luciferase,equipped with a polyhistidine affinity tag,was purified by immobilized metal ion affinity chromatography (IMAC). The approach generated an abundant expression and an efficient purification of a recombinant luciferase with final yield 1.995mg/L of cell culture. Experiments on the recombinant luciferase also showed that the relative light units (RUL) of the enzyme were 5.8×108,and the specific activity was 2.9×1010 RLU/mg. By applying adenosine triphosphate (ATP) bioluminescence to detection of the coin bacteria using the recombinant protein,the ATP content of bacteria was 9.48×10-16mol/mL,and was identical to the bacteria counts (4500CFU/mL) in order of magnitude. Taken together,our results provided a simple and efficacious method of the preparation of recombinant luciferase,which could be applied in the determination of bacteria via ATP bioluminescence.

关键词： ATP bioluminescence bacterial detection expression firefly luciferase purification

Primary study of muscone's effect on cardiovascular system

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沈阳药科大学学报 2008年第S1期25卷 93-94页

作者： ZHU Xue-jing1,2.WU Qi-biao1,LI Hai-tao1,2.1.college of Traditional Chinese Medicine,Macao University of Science and technology,Macao 00853,{2. 2.college of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 2.0046,{2.)

Objective To investigate the effect of muscone on cardiovascular *** Experimental animals to divide muscone high、middle、low dose group(the mouse is 2. mg·kg-1,10 mg·kg-1,5.0 mg·kg-1;the rat is 10 mg·kg-1,5.0 mg·kg-1,2.5 mg·kg-1),GT group(the mouse is 1/12.mg·kg-1;the rat is 1/2. mg·kg-1)and NS *** administration in a week,do the mouse ant-hypoxia experiment、the drug(Pit.)produce the rat myocardial ischemia experiment and obstruct coronary artery to produce the rat myocardial ischemia *** mice's survival time(t),the rat's variation of T in electrocardiogram、creatinkinase(CK)and lactate dehydrogenase(LDH)were recorded,*** The effect of Muscone is significant difference between GT and NS in a dose variation *** Muscone has the effect of ant-hypoxia,cutting down T peak value,reducing CK and *** muscone has effect to inhibiting myocardial ischemia.

关键词： muscone cardiovascular system

东方鲎Factor C中的结构域在结合脂多糖中的作用

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生物化学与生物物理进展 2004年第8期31卷 736-740页

作者：孙向军王东宁张惟杰吴祥甫上海交通大学农业与生物技术学院上海201101 上海交通大学生命科学与技术学院上海200030 中国科学院上海生命科学研究院生物化学与细胞生物学研究所上海200031

FactorC是鲎血细胞中的一种丝氨酸蛋白酶原 ,因其能以高亲和力结合脂多糖 (LPS) ,在医药产品的内毒素检测中起着重要的作用 .以往研究表明处于N端的 3个Sushi结构域对FactorC的LPS结合活性起着关键作用 ,而FactorCN端的其他 3个结构域 ... 详细信息

FactorC是鲎血细胞中的一种丝氨酸蛋白酶原 ,因其能以高亲和力结合脂多糖 (LPS) ,在医药产品的内毒素检测中起着重要的作用 .以往研究表明处于N端的 3个Sushi结构域对FactorC的LPS结合活性起着关键作用 ,而FactorCN端的其他 3个结构域 ,包括Cys rich结构域、EGF like结构域和Lectin like结构域对LPS结合活性的影响尚不清楚 .利用Bac to Bac昆虫细胞表达系统可使rFactorC及其 4个截短的片段rCES12.3L ,rCES12.3,rS12.3L和rS12.3获得表达 ,并且可采用亲和层析柱将这 5个重组表达产物纯化 .通过测定 5个重组表达产物的LPS结合活性及抑菌活性 ,可以确定FactorC的LPS结合位点位于S12.3区域 .尽管Cys rich结构域、EGF like结构域和Lectin like结构域中不存在LPS结合位点 ,但当这 3个结构域同时存在时 ,可提高FactorC或CES12.3L的LPS结合能力 ,因此rCES12.3L具有与rFactorC非常相近的LPS结合能力 .实验结果表明 ,rCES12.3L在昆虫细胞中的表达量比rFactorC高出 4倍 ,预示出rCES12

关键词：东方鲎血细胞丝氨酸蛋白酶原体外抑菌活性细菌内毒素脂多糖

The expert system of genotype discrimination for D5S818 locus based on near-infrared spectroscopy-principal discriminant variate

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Journal of pharmaceutical Analysis 2012年第2期2卷 117-122页

作者： Zai-Zhen Wu a,Jian-Hua Tang a,Bin Zhang a,Li-Ping Guo b,Hong-Ping Xie a,Bing-Ren Gu c a college of pharmaceutical sciences,Soochow University,Suzhou 2.512.,china b {2. of Chemistry and Chemical Engineering,Chongqing University of Science and technology,Chongqing 401331,china c Suzhou Institute for Drug Control,Suzhou 2.5002.china college of pharmaceutical sciences Soochow University Suzhou 215123 China. college of Chemistry and Chemical Engineering Chongqing University of Science and Technology Chongqing 401331 China. Suzhou Institute for Drug Control Suzhou 215002 China.

This paper studied the expert system of genotype discrimination for the STR locus D5S818 based on near-infrared spectroscopy-principal discriminant variate (PDV).Six genotypes,*** 10-10,10-11,11-11,11-12.11-13 and 13-13,were selected as research *** on the optimum polymerase chain reaction (PCR) conditions,about 54 measuring samples for each genotype were obtained;these samples were tested by near-infrared spectroscopy *** differences between homozygote genotypes and heterozygote ones,and differences of the total number of core repeat units between the six genotypes,two types of genotyping-tree structure were constructed and their respective PDV models were studied using the near-infrared spectra of the samples as recognition ***,based on the classification ability of these two genotyping-tree structures,an optimum expert system of genotype discrimination was built using the PDV *** result demonstrated that the built expert system had good discriminability and robustness;without any preprocessing for PCR products,the six genotypes studied could be discriminated rapidly and *** provided a methodological support for establishing an expert system of genotype discrimination for all genotypes of locus D5S818 and other STR loci.

关键词： Short tandem repeat Near-infrared spectroscopy Principal discriminant variate Genotyping-tree structure Expert system

Evolution of binding domain in charge of attachment to heparan sulfate proteoglycan(HSPG)on cell surface may be an important factor affecting formation of some HPV types

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Evolution of binding domain in charge of attachment to hepar...

Inosine inhibits apoptosis and cytochrome C mRNA expression in rat neurons after cerebral ischemia/reperfusion

重庆市生物化学与分子生物学学术会议

作者： Yuegang Zhou (college of pharmaceutical sciences and Bioengineering,Chongqing University of technology, Chongqing 400050,P.R.{2.)

The key amino acid residues,K,K and K,of the binding domain in charge of attachment to heparan sulfate chains of proteoglycans(PGs) on cell surface have been confirmed in human papilloma viruses(HPVs)(Knappe et al.,2.07).In this paper,I aimed to estimate relationship between evolution of the binding domain and formation of HPV *** protein sequence data of 96 HPV types were used in this *** of functional conservation of sites with CLUSTALW program showed that

关键词： human papilloma virus heparan sulfate evolution binding domain mutation

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Neural Regeneration Research 2006年第7期1卷 589-592页

作者： Jinrong Wang1, Mingjun Bi1, Qin Li2.1Department of Neurology, Rongcheng Second People’s Hospital, Rongcheng 2.4309, Shandong Province, china 2.epartment of Neurology, Affiliated Union Hospital, Tongji Medical college, Huazhong University of Science and {2., Wuhan 430030, Hubei Province, china Department of Neurology Rongcheng Second People's Hospital Rongcheng 264309 Shandong Province China Department of Neurology Union Hospital Huazhong University of Science ad Technology Wuhan 430030 Hubei Province China

BACKGROUND: It has been demonstrated that adenosine can induce glial cell to release cytochrome C, enhance expression of apoptotic gene bax, inhibit anti-apoptotic gene bcl-2. and activate caspase-3 to apoptosis; Whereas inosine can inhibit neuronal apoptosis which is similar to bcl-2. OBJECTIVE: To observe the effects of inosine on neuronal apoptosis and expression of cytochrome C mRNA in rats after focal cerebral ischemia/reperfusion, and analyze the pathway of its neuroprotective effect. DESIGN: A randomised controlled animal trial. SETTINGS: Department of Neurology, Rongcheng Second People's Hospital; Department of Neurology, Affiliated Union Hospital, Tongji Medical college, Huazhong University of Science and technology. MATERIALS: Sixty-eight rats, weighing 2.0-2.0 g and clean grade, were used. TdT-mediated dUTP-biotin nick end labeling (TUNEL) and cytochrome C mRNA in situ hybridization kits and DAB staining kit were purchased from Wuhan Boster Biological Co., Ltd.; Inosine injection [2.0 mg (2.mL) each] from Qingdao First {2. Factory. METHODS: The experiment was accomplished in the animal experimental center in Tongji Medical college of Huazhong University of Science and technology from December 2.03 to June 2.05. ① Sixty-four rats were made into focal ischemia by middle cerebral artery occlusion (MCAO) with a nylon monofilament suture. The successfully induced rats were assigned to inosine group (n =32. and model group (n =32. at random. Rats in the inosine group were intraperitoneally administrated with inosine in dose of 100 mg/kg preoperatively, twice a day, 7 days in all. The rats in the control group were injected with the same dose of saline solution by the similar way preoperatively. Each group was randomized into ischemia /reperfusion 2. 6, 12. 2. hours, 2. 3, 7 and 14 days subgroups consisted of 4 rats. The other 4 rats were taken as the sham-operated group, the rats were given the same treatment except for not introduced the filament in

关键词： mRNA Inosine inhibits apoptosis and cytochrome C mRNA expression in rat neurons after cerebral ischemia/reperfusion

Key role of 15-LO/15-HETE in pulmonary vascular remodeling and vascular angiogenesis associated with hypoxic pulmonary hypertension

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Key role of 15-LO/15-HETE in pulmonary vascular remodeling a...

咪唑(啉)-2.羧酸盐作为潜在的/可热活化的有机小分子催化剂催化转酯反应(英文)

第八届海峡两岸心血管科学研讨会

作者： Cui Ma~1,Ya-qian Li~1,Da-ling Zhu~(1,2.) (1.Department of biopharmaceutical sciences,{2. of Pharmacy,Harbin Medical University,Nangang District,Harbin,Heilongjiang Province,china 2.Bio-pharmaceutical Key Laboratory of Heilongjiang Province,Harbin,china)

Objective We have found that 15 - hydroxyeicosatetraenoic acid(15 - HETE) induced by hypoxia was an important mediator in regulation of hypoxic pulmonary hypertension,including the pulmonary vasoconstriction and ***,the underlying mechanisms of the remodeling induced by 15 - HETE are poorly *** In this study,we performed immunohistochemistry,pulmonary artery endothelial cells(PAECs) migration and tube formation in vitro and angiogenesis in vivo,pulmonary artery smooth muscle cells(PASMCs) Bromodeoxyuridine incorporation,cell cycle and DNA analysis to determine the role of 15 - HETE in hypoxia - induced pulmonary vascular remodeling(PVR).Results We found that hypoxia induced morphological changes of pulmonary vessels,including medial hypertrophy and intimal endothelial cells migration and angiogenesis,which were mediated by 15 - ***,15 - HETE stimulated tube formation and migration of PAECs in vitro and angiogenesis in vivo,and signifcantly increased PASMCs ***,15 -HETE regulated the cell cycle progression and made more PASMCs from the G/G phase to G/M + S phase,and hypoxia enhanced the microtubule formation in cell nucleus,which was regulated by 15 - LO/15 - *** addition,we found that Rho - kinase(ROCK) pathway was involved in 15 - HETE - induced PAECs migration and PASMCs *** These results show that 15 - HETE mediates hypoxia - induced PVR and stimulates angiogenesis via ROCK pathway which may be a important mechanism underlying the treatment of pulmonary hypertension.

关键词：

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催化学报 2012年第3期33卷 502-507页

作者：王燕芹李振江南京工业大学生物与制药工程学院材料化学工程国家重点实验室江苏南京210009 南京林业大学理学院江苏南京210037

采用咪唑(啉)-2.羧酸盐(ImCO2.作为催化剂前驱体,通过温度控制脱除ImCO2.2.位羧酸根,再原位释放出高活性的N-杂环卡宾(NHCs),进而催化转酯反应.研究了NHCs在苯甲酸乙酯与一元醇、二元醇及氨基醇反应体系中的催化活性.结果表明,在该体... 详细信息

采用咪唑(啉)-2.羧酸盐(ImCO2.作为催化剂前驱体,通过温度控制脱除ImCO2.2.位羧酸根,再原位释放出高活性的N-杂环卡宾(NHCs),进而催化转酯反应.研究了NHCs在苯甲酸乙酯与一元醇、二元醇及氨基醇反应体系中的催化活性.结果表明,在该体系中不需要选用活化酯为原料,无需加入分子筛或加大醇酯比提高产率,仅用少量催化剂(0.5mol%),在较短时间内(3h),即可使反应转化率达到95%.

关键词： N-杂环卡宾有机小分子催化剂咪唑(啉)-2.羧酸盐转酯反应非活性酯

Cross-linked Polyethylenimine as Potential DNA Vector for Gene Delivery with High Efficiency and Low Cytotoxicity

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Acta Biochimica et Biophysica Sinica 2006年第11期38卷 780-787页

作者： Wei DONG~(1,2. Guang-Hui JIN~1 Shu-Feng LI~(1,3) Qi-Ming SUN~1 Ding-Yuan MA~1 Zi-Chun HUA~1 1 State Key Laboratory of pharmaceutical Biotechnology,{2. of Life Science,Nanjing University,Nanjing 2.0093,china 2.School of Chemical Engineering Nanjing University of Science and technology,Nanjing 2.0094,china 3 {2. of Basic Medical Science,Southeast University,Nanjing 2.0009,china StateKeyLaboratoryofpharmaceuticalBiotechnology CollegeofLifeScienceNanjingUniversityNanjing210093China SchoolofChemicalEngineering NanjingUniversityofScienceandTechnologyNanjing210094China

Polyethylenimine (PEI) has been known as an efficient gene carrier with the highest cationiccharge *** transfection efficiency of PEI,along with its cytotoxicity,strongly depends on itsmolecular *** enhance its gene delivery efficiency and minimize cytotoxicity,we have synthesizedsmall cross-linked PEI with biodegradable linkages and evaluated their transfection efficiencies in *** study,branched PEI with a molecular weight of 800 Da was cross-linked by small diacrylate[1,4-butanediol diacrylate or ethyleneglycol dimethacrylate (EGDMA)] for 2.6 *** efficiencies of thecross-linked PEI in in vitro transfection of plasmid DNA containing enhanced green fluorescent protein(EGFP) reporter gene were assessed in melanoma B 16F10 cell line and other cell *** cytometrywas used to quantify the cellular entry efficiency of plasmid and the transgene expression *** of the cross-linked PEI in these cells were evaluated by MTT ***-PEI 800-4h,atypical cross-linked PEI reported here,mediated a more efficient expression of reporter gene than thecommercially available 2.-kDa branched PEI control,and resulted in a 9-fold increase in gene deliveryin B16F10 cells and a 16-fold increase in 2.3T cells,while no cytotoxicity was found at the optimizedcondition for gene ***,the transfection activity of polyplexes was preserved in thepresence of serum proteins.

关键词： polyethylenimine gene delivery cytotoxicity B16F10 2.3T cell line

A triterpene isolated from Ganodema lucidum inhibits migration and invasion in breast cancer cells

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A triterpene isolated from Ganodema lucidum inhibits migrati...