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	1. Design，Preparation and in vitro Bioactivity of Monopegylated Recombinant Hirudin
	Hou Beibei ,Li Shirong ,Li Xiaohui ,Xiu Zhilong
	Biology 28 November 2006
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	Abstract:Hirudin is the most potent inhibitor of thrombin found in nature. As a promising anticoagulant, a significant drawback which significantly inhibits its clinical application, is the short serum half-life. In order to prolong circulation half-life in vivo, PEGylation is an effective and commonly used method. In this study Succinimidyl carbonyl mPEG (SC-mPEG) 20kDa was attached to recombinant hirudin (r-Hir) at mildly acidic pH to favor the formation of mono- PEGylated r-Hir, since histidine residues represent the preferred PEGylation sites under mildly acidic conditions, and there is only one histidine residue, His51, in r-Hir. Because of the high ratio of monopegylated product, the reaction mixture was easily separated by a one-step ion-exchange chromatographic(IEC) separation procedure. The proportion of urethane bonds involving carboxyalkylated histidines was assayed using its sensitivity to neutral hydroxylamine, and about 79.71% of the mono-substituted PEGylated r-Hir was PEGylated
	TO cite this article:Hou Beibei ,Li Shirong ,Li Xiaohui , et al. Design，Preparation and in vitro Bioactivity of Monopegylated Recombinant Hirudin[OL].[28 November 2006] http://en.paper.edu.cn/en_releasepaper/content/9973


	2. Codon optimization, expression and characterization of an internalizing anti-ErbB2 single chain antibody in Pichia pastoris
	Hu Siyi,Li Liangwei,Qiao Jingjuan ,Guo Yujie ,Cheng Liansheng ,Liu Jing
	Biology 13 March 2006
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	Abstract:Anti-ErbB2 antibodies are used as convenient tools in exploration of ErbB2 functional mechanisms and in treatment of ErbB2-overexpressing tumors. When we employed the yeast P. pastoris to express an anti-ErbB2 single chain antibody (scFv) derived from the tumor-inhibitory monoclonal antibody A21, the yield did not exceed 1-2 mg/L in shake flask cultures. As we considered that the poor codon usage bias may be one limiting factor leading to the inefficient translation and scFv production, we designed and synthesized the full-length scFv gene by choosing the P. pastoris preferred codons while keeping the G+C content at relatively low level. Codon optimization increased the scFv expression level 3-5 fold and up to 6-10 mg/L. Northern blotting further confirmed that the increase of scFv expression was mainly due to the enhancement of translation efficiency. Investigation of culture conditions revealed that the maximal cell growth and scFv expression were achieved at pH 6.5-7.0 with 2% casamino acids after 72h methanol induction. Secreted scFv was easily purified (>95% homogeneous product) from culture supernatants in one-step by using Ni2+ chelating affinity chromatography. The yield was approximately 10-15 mg/L. Functional studies showed that the A21 scFv could be internalized with high efficiency after binding to the ErbB2-overexpressing cells, suggesting this regent may prove especially useful for ErbB2-targeted immunotherapy.
	TO cite this article:Hu Siyi,Li Liangwei,Qiao Jingjuan , et al. Codon optimization, expression and characterization of an internalizing anti-ErbB2 single chain antibody in Pichia pastoris[OL].[13 March 2006] http://en.paper.edu.cn/en_releasepaper/content/5672


	3. gene organization, alternate splicing and expression pattern of porcine visfatin gene
	Yang Zaiqing,Chen Huaping,Xia Tao ,Zhou Lei ,Chen Xiaodong ,Gan Li ,Yao Wenshu,Peng Yin
	Biology 27 February 2006
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	Abstract:Visfatin is a newly discovered visceral fat-specific adipocytokine. It is upregulated in obesity and exerts insulin-mimetic effects in various tissues in human and mouse. We reported here the cloning and characterization of porcine visfatin, its three alternate splicing variants. Sequence analysis indicated that variant 1 is the predominant form among species, which contains an open reading frame of 1473 bp encoding a 52-kDa protein of 491 amino acids. While the other two variants were predicted to encode two 3’truncated proteins due to early termination. The nucleotide and amino acid sequences deduced from variant 1 were conservative across species. The porcine visfatin gene was composed of eleven exons at least and had exactly the same exon/intron structure as the human orthologs. Nested PCR showed that variant 1 and variant 3 were ubiquitously expressed in porcine tissues and that variant 2 was expressed in most tissues examined with exception of testis and liver. The discovery of the three variants of visfatin in porcine would be useful to the further investigation of the function of the visfatin gene.
	TO cite this article:Yang Zaiqing,Chen Huaping,Xia Tao , et al. gene organization, alternate splicing and expression pattern of porcine visfatin gene[OL].[27 February 2006] http://en.paper.edu.cn/en_releasepaper/content/5388


	4. Ginkgo Biloba Extract (GBE) Enhances Glucose Tolerance in Hyperinsulinism-induced Hepatic Cells
	Yang Zaiqing,Zhou Lei,Xia Tao,Gan Li,Chen Xiaodong
	Biology 23 February 2006
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	Abstract:Ginkgo biloba, an herbal medication, is capable of dropping glucose, fat and lipid peroxide and preventing atherosclerosis and complications in diabetic patients. In our studies, we tested the hypothesis that ginkgo biloba extract (GBE) prevents glucose intolerance induced by hyperinsulinism in hepatocytes. We investigated the effects of GBE on glucose ingestion, glucokinase activity and mRNA levels of key genes in glucose metabolism and insulin signaling pathway. To better show its efficacy, we included a control group that was treated with rosiglitazone, a kind of thiazolidinediones (TZDs). The data showed that GBE repressed glucose ingestion in normal status, whereas it dramatically improved glucose tolerance in insulin resistance status. Moreover, by analyzing gene expression, we suggested that GBE chiefly exerted its effects by stimulating IRS-2 transcription. It should be noted that, not like rosiglitazone, GBE didn’t stimulate overmuch glucose uptake in improving glucose tolerance. It is said that GBE treatment could avoid drug-induced obesity. The data suggested that GBE had potential efficacy to prevent insulin resistance induced by hyperinsulinism.
	TO cite this article:Yang Zaiqing,Zhou Lei,Xia Tao, et al. Ginkgo Biloba Extract (GBE) Enhances Glucose Tolerance in Hyperinsulinism-induced Hepatic Cells[OL].[23 February 2006] http://en.paper.edu.cn/en_releasepaper/content/5331


	5. Isolation and Purification of ATP Synthase in Mitochondria
	zhujie,sunrunguang
	Biology 19 April 2005
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	Abstract:In the past 50 years, scientists have done so much on ATP synthase’s structure and its function already, but people known function more and the mechanism fewer, and there are much more work need to do. High purified ATPase and subunits which should come firstly are the key factors in study of enzyme’s structure and catalytic mechanism. This review has summarized some valuable methods in extraction and purification of ATP synthase through compared with kinds of methods came from different workgroups all of the world , and mainly introduced the isolation of bovine heart mitochondria, submichondrial particles, F1-ATPase, F0-ATPase and their subunits such as a，b ，g，d , e in F1-ATPase and b, c, d, e, f, g, h, F6, A6L in F0 moiety and oligomycin sensitive conferring ATPase (OSCP) and the ATPase inhibitor (IF1) in details.
	TO cite this article:zhujie,sunrunguang. Isolation and Purification of ATP Synthase in Mitochondria[OL].[19 April 2005] http://en.paper.edu.cn/en_releasepaper/content/1886


	6. Influence of conformational changes on the kinetics behavior in hydrolysis of cellulose I and cellulose II¾ Acid Vs Enzymatic
	Wu Bin,Wang LuShan ,Zhao Yue,GAO Peiji
	Biology 07 March 2005
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	Abstract:Cellulose II prepared by mercerizing cotton fibers (cellulose I) is thought to be a useful polymer. Studies show that the hydrolysis of cellulose II by dilute acid is more remarkable than that of cotton fibers, whereas it is as difficult to be hydrolyzed by cellulase as cotton fibers. According to the structural changes such as swell of fibrils and decrease of Cr I % (Crystallinity index) confirmed by Atom force Microscopy (AFM, Atom force Microscopy), X-ray diffraction analysis and FT-IR (Transform-infrared spectrum) spectral analysis, cellulose II should show more susceptibility for both acid and enzymatic hydrolysis. However the crystal structure in cellulose II is more stable and the potential energy is to be higher than those of cotton fibers, which may be the key factors contributing to the decrease of hydrolysis rate by cellulase.
	TO cite this article:Wu Bin,Wang LuShan ,Zhao Yue, et al. Influence of conformational changes on the kinetics behavior in hydrolysis of cellulose I and cellulose II¾ Acid Vs Enzymatic[OL].[ 7 March 2005] http://en.paper.edu.cn/en_releasepaper/content/1650

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