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Expression of CTB-human Insulin(BA) Fusion Protein in Gynostemma Pentapyhllum Makino Callus Cells and Its Hypoglycemic Effect in Mice

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Chemical Research in Chinese Universities 2010年第5期26卷 773-779页

作者： wang Ya-jun QI Feng-chun HAO Shu-mei wang chun-yi SUI Bo SHENG Jun College of Life Science Jilin University Changchun 130012 P R. China Changchun Institute of Biological Products Changchun 130062 P R. China Changchun University of Science and Technology Changchun 130022 P. R. China Changchun Keygen Biological Products Co. Ltd. Changchun 130021 P R. China Yunnan Agricultural University Kunming 650201 P. R. China

The plant expression vector of choleratoxin B subunit（CTB）-human insulin（BA） fusion protein pBI121/（CTB-BA） was constructed first and then the Gynostemma Pentapyhllum Makino callus cell line that could express CTB-human insulin fusion protein was constructed and its hypoglycemic effect was evaluated in mice. The plant expression vector pBl 121/（CTB-BA） was digested with both BamI and SacI. Agrobacterium tumerfaciens strain LBA4404 was transformed with previously constructed recombinant plasmid pBI121/（CTB-BA） via the freeze thawing method, then CTB-BA gene was integrated to G Pentapyhllum Makino callus cells by co-culturing the cells with the transformed LBA4404 strain. The transformed G Pentapyhllum Makino callus cells were identified by DNA sequence assey and RT-PCR. The expressed product was identified by western-blot and its amount was tested by ELISA kit and its blood sugar decreasing effect was tested in mice. The sequences of synthetic CTB and human insulin genes（BA） were completely identical to those designed. Restriction map proved that the length of gene fragment in- serted into expression vector pBI121 was consistent with that expected. The sequence of genomic DNA of expressed product was completely identical to that designed. The result of RT-PCR was consistent with that expected. The expressed product showed a specific band with a relative molecular mass of 17000 by Western-blot. The human insulin expression amount was 6.03 μIU/mL according to the ELISA result. The animal test showed that only the G Pentapyhllum Makino callus cell line itself showed activity in decreasing the blood sugar of mice, however, the activity of the transformed G Pentapyhllum Makino callus cells was much higher, The plant expression vector pBI121/（CTB-BA） was constructed and expressed in the G Pentapyhllum Makino callus cells successfully for the first time. The trans- formed G Pentapyhllum Makino callus cells showed high activity in decreasing the blood sugar of mice. This stu

关键词： Human insulin peptide without C pcptide Choleratoxin B subunit G Pentapyhllum Makino callus cells

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Effect of Temperature-responsive Solution Behavior of PNIPAM-bPPEOMA-b-PNIPAM on Its Inclusion Complexation with α-Cyclodextrin

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Chinese Journal of Polymer Science 2015年第3期33卷 456-464页

作者： Jun-jie Xiao Xiu-bing Li 王曦 chun-wang yi Sheng-pei Su The Key Lab for Fine Processing of Resources and Advanced Materials of Hunan Province Hunan Normal University National & Local Joint Engineering Lab. for New Petro-chemical Materials and Fine Utilization of Resources Hunan Normal University

The effect of temperature-responsive solution behavior of PNIPAM-b-PPEOMA-b-PNIPAM on its inclusion complexation with α-cyclodextrin was studied. The triblock polymer was prepared by reversible addition-fragmentation chain transfer （RAFT） polymerization and formed inclusion complexes （ICs） after selective threading of the PEO segment of the triblock polymer through the cavities of α-CD units. For comparison, PPEOMA homopolymer was prepared and the inclusion complexation with α-CD was also studied. The ICs were prepared with a-CD when the polymer was in different conformations by changing the temperature, and the formed ICs were characterized by XRD, ^1H-NMR, TGA and DSC. The solutions of the ICs show temperature-responsive clear/turbid transition or fluidic emulsion/gel transition depending on the concentration of the α-CD added, and the stoichiometry determined by ^1H-NMR and TGA indicates that the stoichiometry of EO to α-CD of the resulted ICs increases with increasing of temperature.

关键词： Temperature-responsibility Comblike triblock polymer Inclusion complexes.

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Immune Responses to a Dicistronic Plasmid Expressing HBsAg of Hepatitis B Virus and Interferon-γ

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Chemical Research in Chinese Universities 2007年第4期23卷 437-443页

作者： wang chun-yi QI Feng-chun WU Xiao-juan ZHAO Da-peng LENG Mei SHENG Jun College of Life Science Jilin University Changchun 130021 P. R. China Changchun Institute of Biological Products Changchun 130062 P. R. China

DNA vaccines encoding a viral protein have been shown to induce antiviral immune responses and provide pro-tection against subsequent viral challenge. The present article deals with the efficacy of a DNA vaccine greatly im-proved by the simultaneous expression of HBsAg and interferon-T gene. We constructed a dual expression vector pHIN encoding the HBsAg of Hepatitis B virus and murine IFN-T which are connected with Internal Ribosome Entry Site（IRES）. Mice immunized with this dual expression DNA vaccine exhibited the enhancement of cellular immune response and increased the production of anti-HBV surface antibody, compared with the mice of single gene expres-sion control. Taken together, these results demonstrate that the application of a cytokine gene in a DNA vaccine for-mulation as an adjuvant can improve its immunigenicity.

关键词： DNA vaccine Internal Ribosome Entry Site(IRES) Dicistronic expression vector Immunogenicity

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Immunogenicity and Efficacy of Liposome-encapsulated Influenza Split Vaccine in BALB/c Mice

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Chemical Research in Chinese Universities 2007年第3期23卷 314-318页

作者： QI Feng-chun wang chun-yi wang Ya-jun HANG Xue-mei ZHANG Yan SUI Bo LI Xiao-bo SHENG Jun College of Life Science Jilin University Changchun 130012 P. R. China Changchun Institute of Biological Products Changchun 130062 P. R. China College of Public Health Jilin University Changchun 130012 P. R. China

The cellular immunity of current influenza split vaccine is relatively low. It is necessary to develop a novel vaccine to improve the cellular immunity. The authors of this study prepared liposome-encapsulated influenza split vaccine and tested it in BALB/c mice. The mice were immunized once with 4 μg of haemagglutinin of monovalent A/ New Caledonia/20/99 （H1N1 ） encapsulated with liposomes or the split virus vaccine only through intrastomach injection. In a comparative study, it was observed that the liposome-encapsulated vaccine elicited a higher neutralizing antibedy response, more effectively stimulated spleen cell proliferation, increased cell subsets like CD^4＋ and CD^4＋/ CD8^＋ , and triggered IL-4 and IFN-γ production.

关键词： Liposome Immune effect Influenza vaccine

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Expression of Human Interferon α 2b Gene in Ginseng Cells

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Chemical Research in Chinese Universities 2010年第3期26卷 420-426页

作者： REN Qi wang chun-yi SONG Zhi-ming LIU Dan YU Hai-peng SHENG Jun College of Life Science Jilin University Changchun 130012 P. R. China Changchun Institute of Biological Products Changchun 130062 P R. China Liaoning yisheng Biopharmaceutical Co. Ltd. Shengyang 110131 P. R. China First Affiliated Hospital Jilin University Changchun 130031 P R. China Yunnan Agricultural University Kunming 650201 P.. R. China

The human interferon α 2b（hIFN-α2b） gene was cloned into binary vector pBI121 to obtain plant expression vector pBIFN. The recombinant plasmid pBIFN was transferred into Agrobacterium tumefaciens strain LBA4404. Then the hIFN-α2b gene was introduced into ginseng callus cells via Agrobacterium-mediated transformation and the ginseng cell line carrying hIFN-α2b gene was selected on G418-containing medium. The presence of target gene in transformed cells was confirmed by PCR and RT-PCR. The results indicate that hIFN-α2b gene has been integrated into the ginseng cells＇ genome, with transcription products, hIFN-α2b expressed by the transgenic ginseng cells was detected by Western blot. It was shown that a specific protein band at 19000 could be observed. Cytopathic effect（CPE） inhibition assay using the W1SH-VSV system shows that the mean antiviral activity of expressed hlFN-a2α was 6.0× 10^4 IU/mL.

关键词： Human interferon × 2b Ginseng cell Plant transformation Agrobacterium tumefaciens

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Evaluation of Breeding Programs Combining Genomic Information in Chinese Holstein

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Agricultural Sciences in China 2011年第12期10卷 1949-1957页

作者： CHEN Jun wang Ya-chun ZHANG yi SUN Dong-xiao ZHANG Sheng-li ZHANG Yuan College of Animal Science and Technology/National Engineering Laboratory for Animal Breeding/Key Laboratory of Agricultural Animal Genetics and Breeding China Agricultural University Beijing 100193 P.R.China

Current study adopted gene flow theory and selection index method to compare the breeding efficiency of three breeding plans in the Chinese Holstein cattle using ZPLAN software. Simulated conventional progeny-testing program （PT） and young sire program （YS） were compared with breeding program using genomic selection （GS） taking parameters derived from Chinese Holstein breeding system. The results showed that, GS shortened generation interval by 1.5-2.2 years, and increased the genetic progress by 30-50%, comparing to PT and YS, respectively. Economic analysis showed that GS could obtain a higher breeding efficiency, being 119 and 97% higher than that of PT and YS, respectively; and GS was also powerful in improving functional traits with a low heritability. Main factors affecting breeding efficiency in GS were further discussed, including selection intensity, accuracy and the cost of SNP genotyping. Our finding provided references for future designing and implementing GS in Chinese dairy population.

关键词： Chinese Holstein genomic selection progeny-testing program young sire program breeding plan

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Suppression Subtractive Hybridization Reveals Different Responses of Two Varieties of Gossypium arboreum L. Under Apolygus lucorum Stress

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Journal of Integrative Agriculture 2014年第6期13卷 1250-1257页

作者： ZHANG Shuai LI Jing Lü Li-min wang chun-yi LUO Jun-yu CUI Jin-jie State Key Laboratory of Cotton Biology/Institute of Cotton Research Chinese Academy of Agricultural Sciences

Plants reshape their transcriptomes, proteomes and metabolomes in response to insect damage. In this study, we used suppression subtractive hybridization to investigate the transcriptomes of two cotton varieties （CCRI41 and CCRI23） under Apolygus lucorum damage. From the CCRI23 libraries we obtained 92 transcripts and from the CCRI41 libraries we obtained 96 transcripts. 26 and 63 of the transcripts from CCRI23 and CCRI41, respectively, had known functions. Using reverse transcription PCR, we detected expression proifle of genes with known functions. Ultimately, we identiifed eight signiifcantly regulated genes, including one downregulated and four upregulated genes from the CCRI41 libraries, and one downregulated and two upregulated genes from the CCRI23 libraries. Only the gene encoding the polyphenol oxidase （PPO） is involved in plant defense against insect herbivores, and the others are related to improving tolerance to insect damage. Quantitative real-time PCR was used to study changes in expression levels during A. lucorum damage in CCRI23 and CCRI41. Signiifcantly regulated genes from CCRI23 showed a response in CCRI23 but not response in CCRI41. Similarly, signiifcantly regulated genes from CCRI41 showed a response in CCRI41 but not response in CCRI23. The results showed that, among transcriptomes of cotton varieties, there are different responses to A. lucorum damage.

关键词： suppression subtractive hybridization Apolygus lucorum defense against insect herbivores

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Effects of soil salinity on rhizosphere soil microbes in transgenic Bt cotton fields

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Journal of Integrative Agriculture 2017年第7期16卷 1624-1633页

作者： LUO Jun-yu ZHANG Shuai ZHU Xiang-zhen LU Li-min wang chun-yi LI chun-hua CUI Jin-jie ZHOU Zhi-guo State Key Laboratory of Cotton Biology Institute of Cotton Research Chinese Academy of Agricultural Sciences College of Agriculture Nanjing Agricultural University/Key Laboratory of Crop Physiology & Ecology in Southern China Ministry of Agriculture

With increased cultivation of transgenic Bacillus thuringiensis （Bt） cotton in the saline alkaline soil of China, assessments of transgenic crop biosafety have focused on the effects of soil salinity on rhizosphere microbes and Bt protein residues. In 2013 and 2014, investigations were conducted on the rhizosphere microbial biomass, soil enzyme activities and Bt protein contents of the soil under transgenic Bt cotton （variety GK19） and its parental non-transgenic cotton （Simian 3） cultivated at various salinity levels （1.15, 6.00 and 11.46 dS m-1）. Under soil salinity stress, trace amounts of Bt proteins were ob- served in the Bt cotton GK19 rhizosphere soil, although the protein content increased with cotton growth and increased soil salinity levels. The populations of slight halophilic bacteria, phosphate solubilizing bacteria, ammonifying bacteria, nitrifying bacteria and denitrifying bacteria decreased with increased soil salinity in the Bt and non-Bt cotton rhizosphere soil, and the microbial biomass carbon, microbial respiration and soil catalase, urease and alkaline phosphatase activity also decreased. Correlation analyses showed that the increased Bt protein content in the Bt cotton rhizosphere soil may have been caused by the slower decomposition of soil microorganisms, which suggests that salinity was the main factor influencing the relevant activities of the soil microorganisms and indicates that Bt proteins had no clear adverse effects on the soil microorganisms. The results of this study may provide a theoretical basis for risk assessments of genetically modified cotton in saline alkaline soil.

关键词： soil salinity Bt cotton soil microorganisms microbial biomass carbon microbial respiration soil enzyme activityBt protein

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The biotypes and host shifts of cotton-melon aphids Aphis gossypii in northern China

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Journal of Integrative Agriculture 2018年第9期17卷 2066-2073页

作者： ZHANG Shuai LUO Jun-yu wang Li wang chun-yi LU Li-min ZHANG Li-juan ZHU Xiang-zhen CUI Jin-jie State Key Laboratory of Cotton Biology Institute of Cotton ResearchChinese Academy of Agricultural SciencesAnyang 455000P.R.China

Aphis gossypfi is a globally distributed species and therefore has a highly variable life cycle. Populations of A. gossypii in northern China exhibit greater genotypic diversity and a broader host range, yet the details of life cycles of different biotypes is still unclear. In this study, the Cytb and 16S gene regions of A. gossypfi collected from 5 common summer hosts and 4 primary hosts were analyzed. A total of 57 haplotypes were obtained from 1 046 individual A. gossyp# sequences. The sequence included 44 variable sites, 27 of which were parsimony informative sites and 17 of which were singleton variable sites. The most frequent 3 haplotypes were found in 896 individuals, representing a total of 85.7% of all individuals and 36 haplotypes were found in 1 individual. A neighbor-joining tree was constructed using 21 haplotypes that were found in more than 2 individuals. Considering the individual host plant, 5 biotypes were identified. Type 1 corresponded exactly to the cucurbit host-race and the other 4 biotypes were found as cotton host-races. Type 3 was the most abundant biotype in cotton fields in northern China.

关键词： cotton-melon aphid Cytb and 16S genes haplotype biotype cotton field

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Neurological deterioration in the acute phase of minor ischemic stroke is an independent predictor of poor outcomes at 1 year： results from the China National Stroke Registry （CNSR）

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Chinese Medical Journal 2013年第18期126卷 3411-3416页

作者： JU yi ZHAO Xing-quan wang chun-xue wang yi-long LIU Gai-fen wang Yong-jun Department of Neurology Beijing Tiantan Hospital Capital Medical University Beijing 100050 China

Background The risk of clinical deterioration still exists in the acute phase despite the fact that patients with minor stroke may display less severe symptoms. The impact of this clinical deterioration on long-term outcomes is unknown. We characterized the clinical features of neurological deterioration （ND） in the acute phase of minor ischemic stroke （MIS） and investigated its impact on mid- and long-term outcomes. Methods This was a multi-centered, prospective clinical study involving patients with MIS （the National Institutes of Health Stroke Scale, NIHSS ~3） recruited from the China National Stroke Registry. Patients were included who had been hospitalized within 24 hours of stroke onset. Baseline characteristics, complication rates during hospitalization, etiology of stroke, as well as 3-, 6-, and 12-month post-stroke outcomes were compared between patients with and without ND during the acute phase. Results A number of 368 （15.2%） out of 2424 patients included in the study exhibited ND in the acute phase. Compared to patients without ND, patients with ND had longer hospital stay, increased rate of baseline diabetes, and multiple complications. Multivariate Logistic regression indicated that ND in acute phase was an independent factor predictive of increased dependence （adjusted odds ratio = 5.20, 95% CI, 3.51-7.70, P 〈0.001） at 12-month post-stroke. Conclusions The risk of ND in the acute phase is high in patients with MIS. ND in the acute phase is an independent predictor for poor outcomes at 12 months post-stroke onset.

关键词： schemic stroke minor stroke predictor acute phase outcomes

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