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Construction of recombinant adenoviral vector carrying human tissue inhibitor of metalloproteinase-1 gene and its expression in vitro

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Hepatobiliary & Pancreatic Diseases International 2005年第2期4卷 259-264页

作者： Dong Xia Lu-Nan Yan Yu Tong Xin-Ping Wang Ming-Man Zhang lan-ying zhao Department of General Surgery West China Hospital Sichuan University Chengdu 610041 China.

BACKGROUND: Overexpression of human tissue inhibitors of metalloproteinase-1 (TIMP-1) may play an antitumor role in inhibiting hepatocellular carcinoma (HCC) growth and progression. The aim of the study was to construct a recombinant adenovirus vector carrying hTIMP-1 cDNA for liver gene therapy and observe its expression in vitro.METHODS: The full-length cDNA of hTIMP-1 was positively cloned into the adenoviral shuttle vector pAdTrack-CMV, and then cotransformed into competent BJ5183 cells with the adenoviral backbone plasmid pAdEasy-1. Thus, a recombinant adenovirus AdhTIMP-1 containing full-length cDNA of hTIMP-1 was generated by homologous recombination in E. coli. AdhTIMP-1 was then packaged and amplified in adenoviral packaging cells, or human embryonic kidney 293 cells. The viral titer was checked by green fluorescent protein (GFP), and the expression of hTIMP-1 in vitro was detected by the techniques of Western blot and RT-PCR.RESULTS: The recombinant adenovirus vector carrying hTIMP-1 was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. The transcription of TIMP-1 mRNA in 293 cells was checked by RT-PCR and TIMP-1 protein could be detected in the culture by Western blot analysis.CONCLUSION: The successful construction of recombinant adenoviral vector carrying human TIMP-1 and the effective expression in vitro has laid a foundation for further study of its antitumor function and may pave the way for future application in liver gene therapy.

关键词： tissue inhibitor of metalloproteinase hepatocellular carcinoma recombinant adenoviral vector gene therapy

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Research proceedings on amphibian model organisms

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Zoological Research 2016年第4期37卷 237-245页

作者： Lu-Sha LIU lan-ying zhao Shou-Hong WANG Jian-Ping JIANG Chengdu Institute of Biology Chinese Academy of Sciences Chengdu 610041 China University of Chinese Academy of Sciences Beijing 100049 China

Model organisms have long been important in biology and medicine due to their specific characteristics. Amphibians, especially Xenopus, play key roles in answering fundamental questions on developmental biology, regeneration, genetics, and toxicology due to their large and abundant eggs, as well as their versatile embryos, which can be readily manipulated and developed in vivo. Furthermore, amphibians have also proven to be of considerable benefit in human disease research due to their conserved cellular developmental and genomic organization. This review gives a brief introduction on the progress and limitations of these animal models in biology and human disease research, and discusses the potential and challenge of Microhyla fissipes as a new model organism.

关键词： Amphibian Model organism Life Science Biomedicine Microhyla fissipes

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Overexpression of TIMP-1 mediated by recombinant adenovirus in hepatocellular carcinoma cells inhibits proliferation and invasion in vitro

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Hepatobiliary & Pancreatic Diseases International 2006年第3期5卷 409-415页

作者： Dong Xia Lu-Nan Yan Jian-Guo Xie Yu Tong Mao-Lin Yan Xin-Ping Wang Ming-Man Zhang lan-ying zhao Department of General Surgery West China Hospital Sichuan University Chengdu 610041 China.

BACKGROUND: Matrix metalloproteinases (MMPs) and its natural tissue inhibitors of metalloproteinases (TIMPs) are involved in cancer progression. This study was undertaken to determine the effects of overexpression of TIMP-1 on human hepatocellular carcinoma (HCC) cell growth, proliferation,and invasion. METHODS: Employing the efficient AdEasy?system, recombinant adenovirus AdTIMP-1 containing full-length cDNA of TIMP-1 was generated by homologous recombination and amplified in 293 cells. Then, human HCC cell line (HepG2) underwent gene transfection to overexpress TIMP-1 (so-called HepG-T cells). The mRNA and protein expressions of TIMP-1 were detected with RTPCR and Western blotting, respectively. The ultrastructure was observed with a transmission electron microscope and the proliferation of HepG-T cells was determined by MTT assay and growth curve. The potential of in vitro invasion was measured with Millicell Chamber. RESULTS: The resulting AdTIMP-1 and HepG-T cells were generated and the expression of TIMP-1 was detected in vitro. The cell proliferation curves and MTT assay showed HepG-T cells’ growth, and proliferation were obviously inhibited. The invasion across Matrigel-coated filters was significantly decreased compared with controls. The suppression rate of HepG-2 cells with AdhTIMP-1 transfection was 50%, and AdhTIMP-1 transfection inhibited by more than 91.6% of the invasion into the Matrigel-coated filter (P<0.01). CONCLUSIONS: TIMP-1 overexpression results in the suppression of proliferative and invasive potential of HepG2 cells in vitro. This study demonstrates the potential role of TIMP-1 as a target for liver cancer gene therapy and has laid a foundation for further study on its anticancer function.

关键词： tissue inhibitor of metalloproteinase hepatocellular carcinoma proliferation invasion gene therapy

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大型货车传动轴总成的强度分析及改进

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机械设计与制造 2013年第4期 56-58,61页

作者：杨震立蓝盈照重庆大学机械传动国家重点实验室重庆400030 重庆重汽远东传动轴有限责任公司重庆400900

大型货车传动轴是货车传动系统中的关键零部件,起着传递发动机功率的重要作用,但也存在着强度不足引发失效和结构笨重的问题。针对这些问题采用SolidWorks软件建立了传动轴总成的三维实体几何模型,基于abaqus软件建立了传动轴总成的有... 详细信息

大型货车传动轴是货车传动系统中的关键零部件,起着传递发动机功率的重要作用,但也存在着强度不足引发失效和结构笨重的问题。针对这些问题采用SolidWorks软件建立了传动轴总成的三维实体几何模型,基于abaqus软件建立了传动轴总成的有限元模型,并进行了强度分析。根据强度分析结果与满足结构安全的要求,提出了结构改进方案,并对改进方案进行了分析计算。结果表明,改进后传动轴总成重量减轻,各零部件应力集中区域减少,结构更加合理,具有重要的参考价值。

关键词：大型货车传动轴总成强度分析结构改进

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Flexible molecules dedicate to release strain of inverted inorganic perovskite solar cell

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Journal of Energy Chemistry 2025年第1期100卷 87-93页

作者： Hongrui Sun Sanlong Wang Pengyang Wang Yali Liu Shanshan Qi Biao Shi ying zhao Xiaodan Zhang Institute of Photoelectronic Thin Film Devices and Technology Renewable Energy Conversion and Storage Center State Key Laboratory of Photovoltaic Materials and CellsNankai University Key Laboratory of Photoelectronic Thin Film Devices and Technology of Tianjin Haihe Laboratory of Sustainable Chemical Transformations Engineering Research Center of Thin Film Photoelectronic Technology of Ministry of Education Collaborative Innovation Center of Chemical Science and Engineering (Tianjin)

The tensile strain in inorganic perovskite films induced by thermal annealing is one of the primary factors contributing to the inefficiency and instability of inorganic perovskite solar cells （IPSCs）,which reduces the defect formation ***,a flexible molecule 5-maleimidovaleric acid （5-MVA） was introduced as a strain buffer to release the residual strain of *** anhydride and carboxyl groups in 5-MVA interact strongly with the uncoordinated Pb2+through Lewis acid-base reaction,thus tightly“pull”the perovskite *** in-between soft carbon chain increased the structural flexibility of CsPbI2.85Br0.15perovskite materials,which effectively relieved the intrinsic internal strain of CsPbI2.85Br0.15,resisted the corrosion of external strain,and also reduced the formation of defects such as VIand *** addition,the introduction of 5-MVA improved crystal quality,passivated residual defects,and narrowed energy level ***,power conversion efficiency （PCE） of NiOxbased inverted IPSCs increased from 19.25% to 20.82% with the open-circuit voltage enhanced from 1.164 V to 1.230 *** release of strain also improved the stability of CsPbI2.85Br0.15perovskite films and devices.

关键词： Inverted inorganic perovskite solar cells Flexible molecules Strain release Crystallization Energy barrier High PCE

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基于动态降阶模型的电力系统非线性电压预测控制

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电力建设 2017年第3期38卷 34-41页

作者：兰晓明王颖赵洪山米增强华北电力大学电气与电子工程学院河北省保定市071003 国网河北省电力公司经济技术研究院石家庄市050021

传统电压控制多采用潮流方程,电压可能在到达事故后稳定运行点前的过渡过程中失稳,因此基于系统的动态模型进行电力系统电压控制十分必要。该文提出了一种基于动态降阶模型的非线性电压预测控制方法。为降低优化计算时间,结合电力系统... 详细信息

传统电压控制多采用潮流方程,电压可能在到达事故后稳定运行点前的过渡过程中失稳,因此基于系统的动态模型进行电力系统电压控制十分必要。该文提出了一种基于动态降阶模型的非线性电压预测控制方法。为降低优化计算时间,结合电力系统的特点对经验Gramian平衡降阶方法加以改进,并应用改进的经验Gramian平衡降阶方法降低电力系统非线性动态模型的维数。为提高模型计算精度和数值稳定性,提出使用4阶收敛的Adams法替代欧拉法进行状态预测,建立基于降阶模型的多步预测-滚动优化模型。此外,在模型求解过程中使用温启动方法和较小的迭代次数限值N^(max)来减少迭代次数。以NewEngland 10机39节点电力系统对所提出的方法进行验证。结果表明,所提出的方法能够提高预测模型的数值稳定性,极大地降低模型求解时间,有利于提前响应系统中可预测的动态变化,维持系统电压稳定。

关键词：电力系统电压控制动态模型模型预测控制(MPC) 模型降阶

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Construction of the recombinant adenovirus vector carrying antisense multidrug resistance gene

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Hepatobiliary & Pancreatic Diseases International 2006年第1期5卷 80-84页

作者： Bo Li, Xing-Hua Gou, Lin Chen, De-Hua Li, Yong-Heng zhao, Lei Han, lan-ying zhao and Jian-ping Gong Department of Hepatobiliary Surgery, Second Affiliated Hospital, Chongqing University of Medical Sciences, Chongqing 400010, China Genetic Engineering Laboratory, Chengdu Diao Group Co. Ltd., Chengdu 610041, China and Department of General Surgery, West China Hospital, Sichuan University, Chengdu 610041, China Department of Hepatobiliary Surgery Second Affiliated Hospital Chongqing University of Medical Sciences Chongqing 400010 China.

BACKGROUND: Multidrug resistance proteins serve as transporters for chemical drugs in human malignancies. The objective of this study was to construct a homologous recombinant adenovirus carrying a reversal fragment of multidrug resistance gene 1 (mdr1) gene cDNA sequence. METHODS: The fragment of the mdr1 gene from the plasmid pHaMDRI-1 carrying the whole human mdr1 cDNA sequence was inserted reversely into the shuttle plasmid pAdTrack-CMV of adenoviral vector system AdEasy. The homologous recombination process was taken place in E. coli BJ5183 with the backbone plasmid pAdEasy-1. After packaging in 293 cells, recombinant adenoviral plasmid was generated. The recombinant adenoviral plasmid was identified by polymerase chain reaction (PCR), restriction endonucleases digest, DNA sequence analysis and fluorescence microscopic photograph, respectively. RESULTS: The recombinant adenovirus pAdEasy-GFPASmdr1 was successfully constructed and identified by PCR, restriction digest, and sequencing with strong green fluorescence expression in fluorescence microscopic photograph. CONCLUSIONS: The recombinant adenoviral mdr1 vector would introduce the antisense mdr1 gene into the human multidrug resistance hepatocellular cell fine effectively, which would provide an experimental basis to study the multidrug resistance in human hepatocellular carcinoma.

关键词： antisense RNA technique multidrug resistance recombinant adenovirus

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Storage time affects the level and diagnostic efficacy of plasma biomarkers for neurodegenerative diseases

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Neural Regeneration Research 2025年第8期20卷 2373-2381页

作者： Lifang zhao Mingkai Zhang Qimeng Li Xuemin Wang Jie Lu ying Han Yanning Cai Department of Clinical Biobank Xuanwu HospitalCapital Medical UniversityBeijingChina Beijing Geriatric Medical Research Center BeijingChina Department of Neurology Xuanwu HospitalCapital Medical UniversityBeijingChina Department of Radiology and Nuclear Medicine Xuanwu HospitalCapital Medical UniversityBeijingChina Beijing Key Laboratory of Magnetic Resonance Imaging and Brain Informatics BeijingChina Key Laboratory of Neurodegenerative diseases Ministry of EducationBeijingChina School of Biomedical Engineering Hainan UniversityHaikouHainan ProvinceChina Center of Alzheimer’s Disease Beijing Institute for Brain DisordersBeijingChina National Clinical Research Center for Geriatric Diseases BeijingChina Institute of Biomedical Engineering Shenzhen Bay LaboratoryGaoke Innovation CenterShenzhenGuangdong ProvinceChina Department of Neurobiology Xuanwu HospitalCapital Medical UniversityBeijingChina

Several promising plasma biomarker proteins,such as amyloid-β(Aβ),tau,neurofilament light chain,and glial fibrillary acidic protein,are widely used for the diagnosis of neurodegenerative ***,little is known about the long-term stability of these biomarker proteins in plasma samples stored at-80°*** aimed to explore how storage time would affect the diagnostic accuracy of these biomarkers using a large *** samples from 229 cognitively unimpaired individuals,encompassing healthy controls and those experiencing subjective cognitive decline,as well as 99 patients with cognitive impairment,comprising those with mild cognitive impairment and dementia,were acquired from the Sino Longitudinal Study on Cognitive Decline *** samples were stored at-80°C for up to 6 years before being used in this *** results showed that plasma levels of Aβ42,Aβ40,neurofilament light chain,and glial fibrillary acidic protein were not significantly correlated with sample storage ***,the level of total tau showed a negative correlation with sample storage ***,in individuals without cognitive impairment,plasma levels of total protein and tau phosphorylated protein threonine 181(p-tau181)also showed a negative correlation with sample storage *** was not observed in individuals with cognitive ***,we speculate that the diagnostic accuracy of plasma p-tau181 and the p-tau181 to total tau ratio may be influenced by sample storage ***,caution is advised when using these plasma biomarkers for the identification of neurodegenerative diseases,such as Alzheimer's ***,in cohort studies,it is important to consider the impact of storage time on the overall results.

关键词： Alzheimer’s disease amyloid-β diagnostic ability glial fibrillary acidic protein neurodegeneration neurofilament light chain plasma biomarkers single molecule array storage time tau

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华东地区夏季极端高温特征及其对植被的影响

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中国农业气象 2017年第1期38卷 42-51页

作者：葛非凡毛克彪蒋跃林谭雪兰赵映慧夏浪安徽农业大学资源与环境学院合肥230036 中国农业科学院农业资源与农业区划研究所/呼伦贝尔草原生态系统国家野外科学观测研究站北京100081 湖南农业大学资源环境学院长沙410128 东北农业大学资源与环境学院哈尔滨150030 北京农林科学院国家农业智能装备工程技术研究中心北京100097

基于中国华东地区79个气象站1971-2006年夏季逐日地面最高气温和1981-2006年先进超高分辨率辐射计(AVHRR)的归一化植被指数(NDVI)资料,分析华东地区夏季极端高温热浪频次和平均持续时间的空间分布特征,并利用Morlet小波变换分析极端高... 详细信息

基于中国华东地区79个气象站1971-2006年夏季逐日地面最高气温和1981-2006年先进超高分辨率辐射计(AVHRR)的归一化植被指数(NDVI)资料,分析华东地区夏季极端高温热浪频次和平均持续时间的空间分布特征,并利用Morlet小波变换分析极端高温次数的周期性变化规律,同时采用奇异值分解(SVD)研究华东地区极端高温次数与植被指数之间的联系。结果表明:(1)华东地区夏季极端高温热浪频次的空间分布主要以北低南高,东低西高,平原高山区低为主。平均持续时间相对于频次,其高值区更靠近沿海。(2)华东地区夏季极端高温次数主要受22a、9a、4a左右的尺度波动影响。其中22a左右的时间尺度为第一主周期,周期振荡在整个时域中表现较稳定。第二主周期为9a左右,周期振荡的振幅从1994年开始逐渐由小增大,将对未来产生重要影响。(3)华东地区夏季极端高温次数与NDVI存在显著相关。当华东中南部地区夏季极端高温次数偏高时,江苏东部地区的夏季植被覆盖度偏低,而山东北部地区和江西西南及西北地区的夏季植被覆盖度偏高;当山东中部地区夏季极端高温次数偏低时,山东东北地区的夏季植被覆盖度偏高,而江苏东部、福建南部和江西北部的夏季植被覆盖度偏低。

关键词：极端高温小波周期奇异值分解植被

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Effects of multidrug resistance, antisense RNA on the chemosensitivity of hepatocellular carcinoma cells

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Hepatobiliary & Pancreatic Diseases International 2006年第4期5卷 552-559页

作者： Bo Li, Tian Ye, Lei zhao, De-Hua Li, Xing-Hua Gou, lan-ying zhao, Lei Han, Lin Chen, Lu-Nan Yan and Jian-Ping Gong Department of Hepatobiliary Surgery, Second Affiliated Hospital, Chongqing University of Medical Sciences, Chongqing 400010, China Department of Traditional Chinese Surgery, Second Affiliated Hospital, Luzhou Medical College, Luzhou 646000, China Genetic Engineering Laboratory, Chengdu Di’ao Group Co., Ltd., Chengdu 610041, China Department of General Surgery, West China Hospital, Sichuan University, Chengdu 610041, China Department of Hepatobiliary Surgery Second Affiliated Hospital Chongqing University of Medical Sciences Chongqing 400010 China.

BACKGROUND: Multidrug resistance is a major obstacle in cancer chemotherapy. We examined whether the antisense RNA of multidrug resistance gene 1 (mdr1) could reverse multidrug resistance in the human hepatocellular carcinoma (HCC) cell line SMMC7721/ADM. METHODS: The recombinant adenoviruses pAdEasy- GFP-ASmdr1 product was produced by the adenoviral vector AdEasy system, which can express antisense RNA against the mdr1 gene. Following that, the recombinant adenovirus was transfected into the P-glycoprotein- producing multidrug resistance cell line, SMMC7721/ADM human HCC cells resistant to adriamycin (ADM) and daunorubicin (DNR). In order to investigate the reversal of multidrug resistance phenotype, we measured the expression of mdr1 mRNA by RT-PCR and the production of P-glycoprotein by flow cytometry. The sensitivities for ADM and DNR SMMC7721/ADM cells were examined by [3-(4, 5-dimethylthi-azol-2-yl)-2,5 diphenyl-terazolium bromide] (MTT) analysis. RESULTS: The low-level expression of mdr1 mRNA and P-glycoprotein production were observed in parental sensitive cells SMMC/7721 in addition to the overexpressionof mdr1 mRNA and P-glycoprotein in SMMC7721/ADM cells. The transfection of antisense-RNA into SMMC7721/ ADM cells resulted in decreases of mdr1 mRNA and P-glycoprotein, but increase of drug sensitivities. The sensitivities of transfected SMMC7721/ADM cells to ADM and DNR in IC50 reduced by 31.25% and 62.96% respectively. CONCLUSIONS: Mdr1 antisense RNA can increase the sensitivities of SMMC7721/ADM cells to anticancer drug by decreasing the expression of the mdr1 gene and inhibiting P-glycoprotein expression. This strategy may be applicable to cancer patients with P-glycoportein mediated multidrug resistance.

关键词： antisense nucleic acid multiple drug resistance P-glycoprotein

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