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Effect of Excipients on Stability and Structure of rhCuZn-SOD Encapsulated in PLGA Microspheres

Chemical Research in Chinese Universities 2004年第3期20卷 323-327页

作者： LIU Ling 1,2.,GE Yu 1 and YUAN Qin-sheng 1 1. State Key Laboratory of Bioreactor Engineering and institute of Biochemistry,East china University of Science and Technology,Shanghai 2.02.7,P. R. china 2. Public Health School,Nanjing medical University,Nanjing 2.002.,P. R. china E china Univ Sci & Technol State Key Lab Bioreactor Engn Shanghai 200237 Peoples R China E china Univ Sci & Technol Inst Biochem Shanghai 200237 Peoples R China Nanjing Med Univ Sch Publ Hlth Nanjing 210029 Peoples R China

When a protein is encapsulated into poly( DL -lactide-co-glycolide)(PLGA) microspheres by means of the double-emulsion method,the harsh microspheres formation process including ultrasonification,exposure to an organic solvent and a polymer may cause the denaturation of the protein. In this study,we investigated the enzymatic activity change and the effect of the excipients on the stability of recombinant human Cu,Zn-superoxide dismutase(rhCu,Zn-SOD) during the emulsification. The specific activity recovery was found to be concentration dependent and the excipients involved such as PEG 600 and Tween 2.,and trehalose were shown to increase the stability of rhCu,Zn-SOD. The protein structural integrity within the microspheres was analyzed by FTIR. The structure of rhCu,Zn-SOD within PLGA microspheres containing trehalose was found to be similar to that of the native solid state,whereas the protein encapsulated during the preparation in the absence of any excipient changed due to the possible hydrophobic interaction with the polymer. The results suggest that a rational stability strategy for protein to be encapsulated into microspheres should aim at different processes.

关键词： Poly( DL -lactide-co-glycolide)(PLGA) microsphere Recombinant human Cu Zn-superoxide dismutase(rhCu Zn-SOD) Fourier transform infrared(FTIR) spectroscopy Protein stability Excipient

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Preparation of Immobilized ε-Polylysine PET Nonwoven Fabrics and Antibacterial Activity Evaluation

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Journal of Wuhan University of Technology(Materials Science) 2011年第4期26卷 675-680页

作者：郝丽梅杨荆泉 institute of medical Equipment the Academy of Military Medical Sciences National Bioprotection Engineering Center

A novel antibacterial material （L-PET） was prepared by immobilizing ε-polylysine on polyethylene terephthalate （PET） nonwoven fabrics. Surface modifications of the fabric were performed by using a chemical modification procedure where carboxyl groups were prepared on the PET surface, a coupling agent was grafted, and the ε-polylysine was immobilized. Scanning electron microscopy （SEM） was used to analyze the surface morphology of the fabrics, while the toluidine blue method and X-ray photoelectron spectroscopy （XPS） were used to evaluate the grafting densities. The antibacterial activities of the L-PET were investigated by using the shaking-flask method. The electron micrographs showed that the surface of the blank PET and the modified fabrics did not change. The results of XPS analysis confirmed that ε-polylysine was successfully grafted onto the surface of PET. The results of the antibacterial experiments showed that L-PET fabrics had excellent antibacterial activity against Escherichia coli and Staphylococcus aureus, and that L-PET fabrics were stable in storage for at least two years.

关键词： PET nonwoven fabric ε-polylysine immobilization antibacterial activity antibacterial material

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Technical development of proteomics

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Chinese Science Bulletin 1999年第16期44卷 1441-1447页

作者： WAN Jinghong and HE Fuchu1.Chongqing medical University, Chongqing 400046, china 2. Beijing {2. of Radiation Medicine, Beijing 100850, china Corresponding author Chongqing medical University Chongqing China Beijing institute of Radiation Medicine Beijing China

Proteome research is an important complementarity for the genome research, which uses quantitative protein-level measurements to characterize biological process. Proteome techniques include two aspects. One is two-dimensional electrophoresis, which is the unique technology to separate the components of proteome. The other is identification of the

关键词： proteome two-dimensional electrophoresis image analysis peptide-mass fingerprinting database.

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Effects of chitosan/collagen substrates on the behavior of rat neural stem cells

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Science china(Life Sciences) 2010年第2期53卷 215-222页

作者： YANG ZhaoYang1,MO LinHong2.DUAN HongMei2.& Li XiaoGuang1,2.1 School of Biological Science and medical Engineering,Beihang University,Beijing 100191,china 2.Beijing {2. for neuroscience,capital medical University,Beijing 100069,china 1. School of Biological Science and medical Engineering Beihang University Beijing 100191 China2. Beijing Institute for Neuroscience Capital Medical University Beijing 100069 China

Spinal cord and brain injuries usually lead to cavity *** transplantation by combining stem cells and tissue engineering scaffolds has the potential to fill the cavities and replace the lost neural *** chitosan and collagen have their unique *** this study,the effects of chitosan and collagen on the behavior of rat neural stem cells (at the neurosphere level) were tested in vitro in terms of cytotoxicity and supporting ability for stem cell survival,proliferation and *** the serum-free condition,both chitosan membranes and collagen gels had low cytotoxicity to *** is,cells migrated from neurospheres,and processes extended out from these neurospheres and the differentiated *** with the above two materials,chitosan-collagen membranes were more suitable for the co-culture with rat neural stem cells,because,except for low cytotoxicity and supporting ability for the cell survival,in this group,a large number of cells were observed to migrate out from neurospheres,and the differentiating percentage from neurospheres into neurons was significantly *** modification of chitosan-collagen membranes may shed light on in vivo nerve regeneration by transplanting neural stem cells.

关键词： chitosan collagen neural stem cells biocompatibility differentiation

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Effects of the CNTF-collagen gel-controlled delivery system on rat neural stem/progenitor cells behavior

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Science china(Life Sciences) 2010年第4期53卷 504-510页

作者： YANG ZhaoYang1, QIAO Hui2.& LI XiaoGuang1,3 1School of Biological Science and medical Engineering, Beihang University, Beijing 100191, china 2.ublic Heath School, Ningxia medical University, Yinchuan 750004, china 3Beijing institute for neuroscience, capital medical University, Beijing 100069, china 1. School of Biological Science and medical Engineering Beihang University Beijing 100191 China2. Public Heath School Ningxia Medical University Yinchuan 750004 China3. Beijing Institute for Neuroscience Capital Medical University Beijing 100069 China

The injury of central nervous system (CNS) usually causes the cavity formation. Although transplantation of neural stem/precursor cells (NSPCs) into the lesioned area of CNS has been shown to be implicated in the functional restoration, the therapeutic result is limited by the poor survival of NSPCs as well as their insufficient proliferation and differentiation abilities. Type-1 collagen is considered as a candidate scaffold or drug delivery system to overcome the aforementioned obstacle. This study observed the effects of the CNTF (ciliary neurotrophic factor)-collagen gel-controlled delivery system and daily addition of soluble-form CNTF on the NSPC survival, migration, proliferation and differentiation. The results showed that, within 12.h of the initial co-culture, CNTF was released in a burst pattern, then the CNTF-collagen gel-controlled delivery system stably released CNTF for up to 12.d. The cell viability test, together with immunohistochemistry, RT-PCR and Western blotting, showed that the CNTF-collagen gel-controlled delivery system supported the NSPCs seeded on the surface of collagen gel survival and facilitated their migration and proliferation. The daily addition of soluble-form CNTF to the medium had similar effects to the CNTF-collagen gel-controlled delivery system, but large quantities of soluble-form CNTF were consumed during the entire process. Taken together, the CNTF-collagen gel-controlled delivery system not only provides a physical scaffold for the transplanted NSPCs to adhere and migrate, but also facilitates the NSPC survival, growth and proliferation, simultaneously reducing the consumption of the expensive growth factors. This system may be used to enhance the microenvironment in the lesioned area of CNS.

关键词： spinal cord neural stem/progenitor cells control release type-1 collagen CNTF

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Correlativity between neural complex and gonadal development in Styela plicata

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Chinese Science Bulletin 1997年第10期42卷 864-867页

作者： FANG Yongqiang, LIN Jiahan and HUANG Weiquan1. Third institute of Oceanography, State Oceanic Administration, Xiamen 361005, china 2. Department of Biology, Xiamen U-niversity, Xiamen 361005, china 3. Fourth Military medical University, Xi’an 710032. china Third institute of Oceanography State Oceanic Administration Xiamen 361005 China

IT has been demonstrated that two reproductive hormones,gonadotropin-releasing hormone(GnRH)and gonadotropin(GTH),exist in the nervous system and Hatschek’s pit oflancelet,a species of Cephalochordata,and that these ... 详细信息

关键词： Styela plicata neural complex reproductive hormone.

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Analysis of a 2. “liquid”plasma lattice

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Science china(Physics,Mechanics & Astronomy) 2004年第1期47卷 129-136页

作者： HUA Jianjun1, LIU Yanhong1 & ZHANG Zhihe2.1. institute of Physics, Chinese Academy of Sciences, Beijing 100080, china 2. College of Bio{2. Engineering, capital University of {2. Sciences, Beijing 100054, china Correspondence should be addressed to Hua Jianjun (email: jj_hua@***) Received September 2., 2.03 1. institute of Physics Chinese Academy of Sciences 100080 Beijing China 2. College of Biomedical Engineering Capital University of Medical Sciences 10054 Beijing China

To study the structural and dynamical properties of a 2. dusty plasma lattice, a set of experiments were performed in an environment of radio frequency discharge. Based on the results of one of the experiments, pair correlation function, bond-orientational correlation function and mean square displacement of such a system were computed and analyzed. The bond-orientational correlation function is found to fit in with the law of e?r , and the mean square displacement is found to increase monotonously with time, both of which, together with the pair correlation function, indicate that the two-dimensional dusty plasma lattice formed in the experiment stayed in a “liquid” state.

关键词： plasma lattice, pair correlation function, bond-orientational correlation function, mean square displacement.

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Tissue-specific expression of GFP reporter gene in germline driven by GATA-2.promoter and enhancers in zebrafish

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Chinese Science Bulletin 2000年第1期45卷 31-34,97页

作者： MENG Anming & LIN Shuo1. Laboratory of Developmental Biology, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, china 2. institute of Molecular Medicine and Genetics, {2. College of Georgia, Augusta, GA 30912. USA Laboratory of Developmental Biology Department of Biological Sciences and Biotechnology Tsinghua University Beijing China institute of Molecular Medicine and Genetics Medical College of Georgia Augusta USA

GATA-2. a transcription factor, is expressed in several types of blood cells and in the central nervous system (CNS), and regulates the differentiation of these cells. We have obtained five zebrafish transgenic germlines that carry and express the green fluorescent protein (GFP) gene ligated to various 5’ flanking sepuences of zebrafish GATA-2.gene. The spatial pattern of GFP expression varies, mainly depending on which regulatory sequence is used, among the germlines. In some of the germlines, the expression of GFP is restricted to the CNS and the enveloping layer (EVL) cells, while in some other lines GFP is observed only in the CNS. it is noted that the intensity of GFP in the transgenic fish remain unchanged after a six-generation passage of the transgenes. The transgenic fish could find its uses in the future in generating tissue-specific, even cellspecific mutant fish and in functional study of related genes through transgenesis.

关键词： zebrafish transgenic GATA-2 GFP germline.

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Applications of homemade kit in mutation detection of genes

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Science china Chemistry 2004年第2期47卷 166-172页

作者： ZHAO Chunxia1, XU Guowang1, SHI Xianzhe1, MA Jianmei2. ZHANG Yan2. L?Shen1,2.& YANG Qing1 1. Dalian institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116011, china 2. Cell Laboratory, Second Affiliated Hospital, Dalian {2. University, Dalian 11602., china 1. Dalian institute of Chemical Physics Chinese Academy of Sciences 116011 Dalian China 2. Cell Laboratory Second Affiliated Hospital Dalian Medical University 116023 Dalian China

Several methods of mutation detection, such as single-strand conformation polymorphism (SSCP), tandem SSCP/heteroduplex analysis and SNaPshot analysis were developed using homemade kit on AB1 310 genetic analyzer, and were successfully applied to mutation detection of 31 colorectal tumor samples. The sieving capability of homemade kit and commercial kit were compared, results demonstrate that homemade kit has higher resolution and shorter analysis time. In clinical tumor samples, 2.% K-ras (exon 1) and 2.% p53 (exons 7–8) were found to have mutations, and all mutations were single point variations. A majority of mutations occurred in one gene, only 1 tumor contained alterations in the two genes, which indicates that development of colorectal cancer lies on alternate pathways, and may correlate with different gene mutations Keywords single nucleotide polymorphism (SNP) - single-strand conformation polymorphism (SSCP) - heteroduplex analysis (HA) - SNaPshot - linear polyacrylamide (LPA) - polydimethylacrylamide (PDMA)

关键词： single nucleotide polymorphism (SNP) single-strand conformation polymorphism (SSCP) heteroduplex analysis (HA) SNaPshot linear polyacrylamide (LPA) polydimethylacrylamide (PDMA)

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Inhibition of β-lactamase-mediated oxacillin resistance in Staphylococcus aureus by a deoxyribozyme

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Acta Pharmacologica Sinica 2007年第11期28卷 1775-1782页

作者： Zheng HOU~2.Jing-ru MENG~2.Jin-rong ZHAO~3,Ben-quan HU~2.Jie LIU~2.Xiao-jun YAN~3,Min JIA~2.Xiao-xing LUO~(2.4)~2.Department of Pharmacology and ~3 institute of Genetic Diagnosis,The Fourth Military medical University,Xi’an 710032.{2. Department of Pharmacology The Fourth Military Medical University Xi'an 710032 China institute of Genetic Diagnosis The Fourth Military Medical University Xi'an 710032 China

Aim:To investigate the oxacillin susceptibility restoration of methicillin-resistantStaphylococcus aureus (MRSA) by targeting the signaling pathway of blaR1-blaZ with a ***:ADNAzyme (named PS-DRz602. targeting blaR1mRNA was designed and *** DRz602.was introduced into a MRSAstrain WHO-2.the colony-forming units of WHO-2.on the Mueller-Hinton agarcontaining 6 mg/L oxacillin and the minimum inhibitory concentrations of oxacillinwere *** inhibitory effects of DRz602.on the expressions of antibi-otic-resistant gene blaR1 and its downstream gene blaZ were detected by realtime ***:PS-DRz602.significantly decreased the transcription ofblaR1 mRNA and led to the significant reduction of blaZ in a concentration-dependent ***,the resistance of S aureus WHO-2.to the β-lactam antibiotic oxacillin was significantly ***:Our resultsindicated that blocking the blaR1-blaZ signaling pathway via DNAzyme mightprovide a viable strategy for inhibiting the resistance of MRSA to β-lactam antibi-otics and that BlaR1 might be a potential target for pharmacological agents com-bating MRSA.

关键词： methicillin-resistant Staphylococcus aureus BlaR1 phosphorothioate deoxyribozyme antibiotic resistance

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