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富含支化精氨酸的两亲性阳离子脂质肽体内外mRNA基因递送能力及安全性评价

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药学学报 2024年第4期59卷 1079-1086页

作者：王一醇蒲艺瑶毕群杰宋相容金蓉蓉聂宇四川大学国家生物医学材料工程技术研究中心/四川大学生物医学工程学院四川成都610064 四川大学生物治疗全国重点实验室四川成都610041

mRNA基因治疗因其可扩展、修饰,不需要进入细胞核及不整合宿主遗传基因等优势而备受关注。在基因治疗中,将mRNA安全有效地递送到细胞内对于基因治疗的成功至关重要。本研究基于前期设计合成的富含支化精氨酸的两亲性阳离子脂质肽(dendri... 详细信息

mRNA基因治疗因其可扩展、修饰,不需要进入细胞核及不整合宿主遗传基因等优势而备受关注。在基因治疗中,将mRNA安全有效地递送到细胞内对于基因治疗的成功至关重要。本研究基于前期设计合成的富含支化精氨酸的两亲性阳离子脂质肽(dendritic arginine&disulfide bond-containing cationic lipopeptide,RLS)基因载体在斑马鱼体内成功实现比商品化试剂Lipofectamine 2000高1.5倍的mRNA转染效果,并通过体外细胞毒性和斑马鱼体内生物安全性研究证实其具有良好的生物安全性。首先,通过核磁共振氢谱(1H NMR)及飞行时间质谱(MALDI-TOF)对阳离子脂质肽的化学成分进行表征。通过动态光散射粒度分析仪(DLS)测试的粒径和电位结果显示,在氮磷(N/P)比为20时,RLS与mRNA复合组装体形成平均粒径约220 nm、表面ζ电位约+21 mV的均匀纳米粒子。体外基因转染中,RLS在人胚肾293细胞(HEK293)和大鼠间充质干细胞(MSC)中的转染效率较Lipofectamine 2000分别提高1.2和3倍。此外,将RLS显微注射至斑马鱼胚胎后,通过评价生存率、孵化率和致畸率等指标,进一步证实了其在体内良好的安全性。综上所述,该富含支化精氨酸的两亲性阳离子脂质肽RLS具有优良的mRNA递送性能和安全性。

关键词：支化精氨酸两亲性阳离子脂质肽基因载体 mRNA递送斑马鱼

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气候变化背景下江苏春茶生长气候适宜性评价

中国农业气象 2024年第10期45卷 1160-1173页

作者：任义方王春乙张旭晖李时睿钱半吨江苏省气候中心南京210041 中国气象科学研究院北京100081 北京师范大学地理科学学部遥感科学国家重点实验室北京100875 溧阳市气象局溧阳213300

利用1981-2020年江苏72个气象站逐日气象资料,基于茶叶春季生长适宜气象条件,应用模糊数学方法,构建日尺度茶叶气候适宜度指数,开展江苏春茶生长气候适宜度等级区域划分和评价,分析气候变化背景下不同年代江苏春茶生长气候适宜度的时空... 详细信息

利用1981-2020年江苏72个气象站逐日气象资料,基于茶叶春季生长适宜气象条件,应用模糊数学方法,构建日尺度茶叶气候适宜度指数,开展江苏春茶生长气候适宜度等级区域划分和评价,分析气候变化背景下不同年代江苏春茶生长气候适宜度的时空变化特征。结果表明:春茶生长气候适宜度时空变化特征各有异同。对于江苏三大茶区来说,春茶生长日尺度气候适宜度均呈单峰型分布形态,高值时段集中于4月上旬-5月上旬,进入高峰期的时间随年代变化呈10d左右的提前趋势。三大茶树主栽区域年尺度气候适宜度指数在2000-2010年达到高峰,且呈上升趋势。各年代间环太湖低山茶区属高适宜区域,连云港茶区属低适宜区域,宁镇扬丘陵茶区主要为中适宜区。江苏春茶生长气候适宜度指数呈北低南高的分布特征,年代际变化特征明显,以0.005·10a-1的速率呈上升趋势,2001-2010年春茶生长气候资源良好。

关键词：气候变化春茶模糊数学气候适宜度江苏

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

Ore Geology,H-O-C Isotopes and 40Ar-39Ar Dating of the Wutonggou Iron Deposit,Eastern Tianshan,NW China:Implications for the Source,Timing,and Genesis of Hydrothermal Mineralization in a Sedimentary Iron Deposit

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Journal of Earth Science 2024年第4期35卷 1170-1185页

作者： chun-Long wang yi-Tian wang State Key Laboratory of Geological Processes and Mineral Resources Collaborative Innovation Center for Exploration of Strategic Mineral ResourcesSchool of Earth ResourcesChina University of GeosciencesWuhan430074China MNR Key Laboratory of Metallogeny and Mineral Assessment Institute of Mineral ResourcesChinese Academy of Geological SciencesBeijing100037China

The Wutonggou iron deposit is located in the well-known iron metallogenic belt in the eastern Tianshan,NW China,and has been regarded as a sedimentary iron *** hydrothermal overprinting could play indispensable roles in the formation of high-grade iron ores in sedimentary iron deposits,previous studies mainly focused on sedimentary-related iron mineralization,while the nature and contribution of hydrothermal fluids are poorly ***,an integrated study of ore geology,H-O-C isotopes and^(40)Ar-^(39)Ar dating,is conducted on the Wutonggou deposit,in order to reveal the features,source,and timing of hydrothermal *** studied deposit includes two mining sections namely the Jianshan and ***δ^(18)O values of early magnetite from the Jianshan section range from+3.0‰to+5.8‰that nearly consistent with classic magmatic magnetite,while increase to 6.3‰-8.0‰in the late *** from the two sections shows comparable H-O isotopic compositions and identical fractionation trends,and is plotted in or periphery to the primary magmatic water *** from the two sections are broadly similar in carbon and oxygen isotopic compositions,and siderite from the Wutonggou section is plotted in the same ***,comparable stable isotopic compositions and evolution trends indicate similar magmatic fluids contributed hydrothermal iron mineralization in the two mining ***,water-rock interactions of varying degrees generated distinct mineralization styles in the Jianshan and Wutonggou sections,and caused the isotopic fractionation in late *** extracted from a hydrothermal siderite ore yielded a^(40)Ar-^(39)Ar plateau age of 299.5±2.0 Ma,indicates the timing of hydrothermal iron mineralization is corresponding to the emplacement of vicinity *** together,the hydrothermal mineralization in the Wutonggou iron deposit was the product of remobilization and upgrading of early sedimentary iron ores,a

关键词： iron deposits H-O-C isotopes 40Ar-39Ar dating sedimentary iron deposits Wutonggou iron deposit eastern Tianshan

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.

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

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

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

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