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Optimized human factor IX expression cassettes for hepatic-directed gene therapy of hemophilia B

null

《医学前沿（英文）》 2015年 第9卷 第1期   页码 90-99 doi: 10.1007/s11684-015-0390-2

摘要：

Gene therapy provides a potential cure for hemophilia B, and significant progress has been achieved in liver-directed gene transfer mediated by adeno-associated viral vectors. Recent clinical trials involving the use of a self-complementary adeno-associated virus serotype 8-human codon-optimized factor IX (AAV8-hFIXco) vector demonstrated encouraging efficacy with hFIX expression stabilized at 1% to 6% of normal level in patients, but safety concerns related to high vector doses are still present. Thus, further improvement of AAV vectors and hFIX expression cassette may positively contribute to the ultimate success of hemophilia B gene therapy. In this study, to obtain a higher expression level of hFIX that potentiates the coagulant capacity of recipients, human FIX expression vector was optimized by upgrading the codon adaption index and adjusting the GC content, inserting a Kozak sequence (GCCACC), and introducing a gain-of-function mutation, R338L (FIX Padua). The efficiency of the published and the presently constructed cassettes was compared through in vivo screening. In addition, the regulatory elements that control the FIX gene expression in these cassettes were screened for liver-specific effectiveness. Among all the constructed cassettes, scAAV-Pre-hFIXco-SIH-R338L, which was the construct under the control of the prothrombin enhancer and prealbumin promoter, resulted in the highest level of coagulant activity, and the expression levels of two constructed cassettes (scAAV-Chi-hFIXco-SIH-R338L and scAAV-Pre-hFIXco-SIH-R338L) were also higher than that of the published cassette (scAAV-LP1-hFIXco-SJ). In summary, our strategies led to a substantial increase in hFIX expression at the protein level or a remarkably elevated coagulant activity. Thus, these reconstructs of hFIX with AAV vector may potentially contribute to the creation of an efficacious gene therapy of hemophilia B.

关键词： factor IX     hemophilia B     liver-specific regulatory elements     hydrodynamic gene transfer    

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Construction of lentiviral vector carrying Rab9 gene and its expression in mouse brain

Youguo HAO, Min ZHANG, Jinzhi XU, Bitao BU, Jiajun WEI

《医学前沿（英文）》 2009年 第3卷 第2期   页码 141-147 doi: 10.1007/s11684-009-0041-6

摘要： Rab proteins and their effectors facilitate vesicular transport by tethering donor vesicles to their respective target membranes. Rab9 mediates late endosome-to- -Golgi-network trafficking. To explore the possibility of Rab9-related gene therapy for neurodegenerative diseases, we packed Lentivirus encoding Rab9. The expressing plasmid pCDH1-MCF1-Rab9-EF1-copGFP was constructed by using molecular biological techniques. The Lentivirus encoding Rab9 cDNA was packed by Lifectamine-2000 mediated co-transfection of the plasmid pPACKH1- , pPACKH1- and pVSV- into 293T cells. DNA sequencing proved the successful construction of pCDH1-MCF1-Rab9-EF1-copGFP. After 72 hours, the expression of GFP could be detected in BV-2 cells. Western blotting revealed that the Rab9 gene expression in BALB/c mice brain was up-regulated significantly 4 weeks after injection with Lentivirus encoding Rab9, which evidenced a satisfactory increasing effect of this virus. Administration of Lenti-Rab9 to postnatal day 3 Niemann-Pick disease type C (NPC) mice reduced motor defects and prevented the weight loss associated with female NPC mice, as well as modulating the death rate of Purkinje neurons. It is concluded that the packaging of Lentivirus encoding Rab9 was successful. Lentivirus encoding Rab9 can increase the expression of Rab9 gene effectively, which might offer a novel means for the treatment of neurodegenerative diseases.

关键词： Rab9     lentivirus     gene therapy     gene transfer    

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Bacteriophages in water pollution control: Advantages and limitations

《环境科学与工程前沿（英文）》 2021年 第15卷 第5期 doi: 10.1007/s11783-020-1378-y

摘要：

•Phages can be better indicators of enteric viruses than fecal indicator bacteria.

关键词： Phage     Water pollution monitoring     Harmful bacteria biocontrol     Horizontal gene transfer     Bacterial resistance    

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Assessment of antibiotic resistance genes in dialysis water treatment processes

Xuan Zhu, Chengsong Ye, Yuxin Wang, Lihua Chen, Lin Feng

《环境科学与工程前沿（英文）》 2019年 第13卷 第3期 doi: 10.1007/s11783-019-1136-1

摘要：

• Quantitative global ARGs profile in dialysis water was investigated.

• Totally 35 ARGs were found in the dialysis treatment train.

• 29 ARGs (highest) were found in carbon filtration effluent.

• erm and mtrD-02 occurred in the final effluent.

• The effluent was associated with health risks even after RO treatment.

关键词： Dialysis water     Treatment process     Antibiotic resistance gene     High-throughput quantitative PCR     Horizontal gene transfer    

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Matrix attachment regions included in a bicistronic vector enhances and stabilizes follistatin gene expressions

Xiaoming HU,Jing GUO,Chunling BAI,Zhuying WEI,Li GAO,Tingmao HU,Shorgan BOU,Guangpeng LI

《农业科学与工程前沿（英文）》 2016年 第3卷 第1期   页码 87-96 doi: 10.15302/J-FASE-2016087

摘要： In the present study, follistatin ( ) gene expression vectors with either a bicistronic gene transfer cassette alone, or a bicistron gene cassette carrying a matrix attachment region (MAR) were constructed and transfected to bovine fetal fibroblasts. Evaluations of both the integration and expression of exogenous indicated that the pMAR-CAG-FST-IRES-AcGFP1-polyA-MAR (pMAR-FST) vector had higher capacity to form monoclonal transgenic cells than the vector without MAR, though transient transfection and integration efficiency were similar with either construct. Remarkably, protein expression in transgenic cells with the pMAR-FST vector was significantly higher than that from the bicistronic vector. Exogenous was expressed in all of the pMAR-FST transgenic mice at F , F and F . Total muscle growth in F mice was significantly greater than in wild-type mice, with larger muscles in fore and hind limbs of transgenic mice. pMAR-FST transgenic mice were also found with more evenly distributed muscle bundles and thinner spaces between sarcolemma, which suggests a correlation between transgene expression-associated muscle development and the trend of muscle growth. In conclusion, a pMAR-FST vector, which excluded the resistant genes and frame structure, enhances and stabilizes gene expressions in both transfected cells and transgenic mice.

关键词： safety of transgenic     bicistron gene transfer body     transgenic mice     muscle development    

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Recent progress in hydrodynamic characteristics research and application of annular centrifugal extractors

《化学科学与工程前沿（英文）》 2022年 第16卷 第6期   页码 854-873 doi: 10.1007/s11705-022-2156-0

摘要： The annular centrifugal extractor (ACE) integrates mixing and separation. It has been widely used in many industrial fields because of its low residence time, compact structure, and high mass transfer efficiency. Most of the literature has focused on flow instabilities, flow visualization, and computational fluid dynamics simulations. More recently, research on hydrodynamic behavior and structural optimization has received widespread attention. With the development of ACE technology, applications have been broadened into several new areas. Hence, this paper reviews research progress regarding ACE in terms of hydrodynamic characteristics and the structural improvements. The latest applications covering hydrometallurgy, nuclear fuel reprocessing, bio-extraction, catalytic reaction, and wastewater treatment are presented. We also evaluate future work in droplet breakup and coalescence mechanisms, structural improvements specific to different process requirements, scaling-up methods, and stability and reliability after scaling-up.

关键词： annular centrifugal extractor     hydrodynamic characteristics     structure optimization     the latest application    

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Invading target cells: multifunctional polymer conjugates as therapeutic nucleic acid carriers

Ulrich L?CHELT, Ernst WAGNER

《化学科学与工程前沿（英文）》 2011年 第5卷 第3期   页码 275-286 doi: 10.1007/s11705-011-1203-z

摘要： Polymer-based conjugates are an interesting option and challenge for the design of nano-sized drug-delivery systems, as they require advanced conjugation chemistry and precise engineering. In the case of nucleic acid therapy, non-viral carriers face several biological barriers during the delivery process, namely 1) protection of the cargo from extracellular degradation, 2) avoidance of non-specific interactions with non-targeted tissues, 3) efficient entry into the target cells, 4) intracellular trafficking to the site of action and 5) cargo release. To take on these obstacles, multifunctional conjugates can act as “smart polymers” with microenvironment-sensing dynamics to facilitate the separate delivery steps. Synthesis of defined polymer architectures with precise functionalization enables structure-activity relationships to be investigated and the integration of key functions for efficient delivery. Thus bioresponsive polymer conjugates, which are equipped with molecular devices responding to the certain microenvironments within the delivery pathway (e.g. pH, redox potential, enzymes) can be assembled. This review focuses on the modular engineering and conjugation of multifunctional polymeric structures for the utilization as “tailor-made” nucleic acid carriers.

关键词： conjugate     DNA     gene transfer     polymer     RNA     targeting    

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Identification of pollution sources in rivers using a hydrodynamic diffusion wave model and improved

《环境科学与工程前沿（英文）》 2023年 第17卷 第7期 doi: 10.1007/s11783-023-1685-1

摘要：

● A hydrodynamic-Bayesian inference model was developed for water pollution tracking.

关键词： Identification of pollution sources     Water quality restoration     Bayesian inference     Hydrodynamic model     Inverse problem    

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Numerical analysis of hydrodynamic process of circular-translational-moving polishing (CTMP)

ZHAI Wenjie, LIU Changxiong, LIANG Yingchun

《机械工程前沿（英文）》 2008年 第3卷 第4期   页码 441-448 doi: 10.1007/s11465-008-0074-6

摘要： By keeping a pad moving relative to a wafer along a circular path without rotation, we developed a polishing technique called circular-translational-moving polishing (CTMP), which permits multidirectional polishing of the work piece and thus bears the advantage of isotropic polishing and a potential increase of material removal rate (MRR) on the wafer. To illuminate the mechanisms of CTMP and determine the optimum process variables in a CTMP process, a three-dimensional hydrodynamic lubrication model for CTMP with a smooth and rigid pad under a quasi-stable state is established in a polar coordinate system. The model equations are further calculated numerically by the finite difference method. The instantaneous distribution of fluid pressure is obtained, which shows that a negative pressure exists. The reason for negative pressure in CTMP and its effect on polishing is discussed. Moreover, the nominal clearance of the fluid film, roll, and pitch angles under different working conditions are obtained in terms of the applied load, moments, and polishing velocity. The obtained numerical analysis can be used as guidance for choosing operation parameters in a practical CTMP application.

关键词： numerical analysis     practical CTMP     circular-translational-moving polishing     CTMP     different    

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Uncertainty quantification of stability and damage detection parameters of coupled hydrodynamic-ground

Nazim Abdul NARIMAN, Tom LAHMER, Peyman KARAMPOUR

《结构与土木工程前沿（英文）》 2019年 第13卷 第2期   页码 303-323 doi: 10.1007/s11709-018-0462-x

摘要： In this paper, models of the global system of the Koyna dam have been created using ABAQUS software considering the dam-reservoir-foundation interaction. Non coupled models and the coupled models were compared regarding the horizontal displacement of the dam crest and the differential settlement of the dam base in clay foundation. Meta models were constructed and uncertainty quantification process was adopted by the support of Sobol’s sensitivity indices considering five uncertain parameters by exploiting Box-Behnken experimental method. The non coupled models results determined overestimated predicted stability and damage detection in the dam. The rational effects of the reservoir height were very sensitive in the variation of the horizontal displacement of the dam crest with a small interaction effect with the beta viscous damping coefficient of the clay foundation. The modulus of elasticity of the clay foundation was the decisive parameter regarding the variation of the differential settlement of the dam base. The XFEM approach has been used for damage detection in relation with both minimum and maximum values of each uncertain parameter. Finally the effects of clay and rock foundations were determined regarding the resistance against the propagation of cracks in the dam, where the rock foundation was the best.

关键词： massed foundation     hydrodynamic pressure     Box-Behnken method     meta model     Sobol’s sensitivity indices    

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Scenario analysis of water pollution control in the typical peri-urban river using a coupled hydrodynamic-water

Haifeng JIA, Shuo WANG, Mingjie WEI, Yansong ZHANG

《环境科学与工程前沿（英文）》 2011年 第5卷 第2期   页码 255-265 doi: 10.1007/s11783-010-0279-x

摘要： The water quality pollution and ecological deterioration in peri-urban rivers are usually serious under rapid urbanization and economic growth. In the study, a typical peri-urban river, Nansha River, was selected as a case study to discuss the scheme of peri-urban river rehabilitation. Located in the north part of the Beijing central region, the Nansha River watershed has been designated as an ecologically friendly garden-style area with high-tech industry parks and upscale residential zones. However, the Nansha River is currently seriously contaminated by urban and rural pollutants from both nonpoint sources (NPS) and point sources (PS). In this study, the pollutant loads from point sources and nonpoint sources in the Nansha River watershed were first assessed. A coupled model, derived from the Environmental Fluid Dynamics Code and Water Quality Analysis Simulation Program, was developed to simulate the hydrodynamics and water quality in the Nansha River. According to the characteristics of the typical peri-urban river, three different PS and NPS control scenarios were designed and examined by modeling analyses. Based on the results of the scenario analysis, a river rehabilitation scheme was recommended for implementation.

关键词： peri-urban river     coupled hydrodynamic-water quality modeling     scenario analysis     river rehabilitation    

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Development of a hydrodynamic model and the corresponding virtual software for dual-loop circulating

Shanwei Hu, Xinhua Liu

《化学科学与工程前沿（英文）》 2021年 第15卷 第3期   页码 579-590 doi: 10.1007/s11705-020-1953-6

摘要： Dual-loop circulating fluidized bed (CFB) reactors have been widely applied in industry because of their good heat and mass transfer characteristics and continuous handling ability. However, the design of such reactors is notoriously difficult owing to the poor understanding of the underlying mechanisms, meaning it has been heavily based on empiricism and stepwise experiments. Modeling the gas-solid CFB system requires a quantitative description of the multiscale heterogeneity in the sub-reactors and the strong coupling between them. This article proposed a general method for modeling multi-loop CFB systems by utilizing the energy minimization multiscale (EMMS) principle. A full-loop modeling scheme was implemented by using the EMMS model and/or its extension models to compute the hydrodynamic parameters of the sub-reactors, to achieve the mass conservation and pressure balance in each circulation loop. Based on the modularization strategy, corresponding interactive simulation software was further developed to facilitate the flexible creation and fast modeling of a customized multi-loop CFB reactor. This research can be expected to provide quantitative references for the design and scale-up of gas-solid CFB reactors and lay a solid foundation for the realization of virtual process engineering.

关键词： multi-loop circulating fluidized bed     mathematical modeling     energy minimization multiscale     virtual fluidization     mesoscale structure    

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人类遗传病的家系收集疾病基因定位克隆与疾病基因功能的研究

夏家辉

《中国工程科学》 2000年 第2卷 第11期   页码 1-11

摘要：

介绍了中国医学遗传学国家重点实验室在遗传病家系收集、疾病基因定位、疾病基因克隆和疾病基因功能研究方面的研究工作。用细胞遗传学G显带技术于1975年发现了一条与鼻咽癌相关的标记染色体t（1；3）（q44；p11）；1981年将睾丸决定基因（TDF）定位于Yp11.32带；1991年以来收集遗传病家系345种共590个；1996年用显微切割、PCR、微克隆技术克隆了EXT2基因；1998年用基因家族-候选疾病基因克隆方法克隆了遗传性神经性耳聋基因GJB3；1999年用连锁分析和全基因组扫描将一种遗传性弥漫性浅表性光敏性汗孔角化症定位于12q23.2带，并在基因功能研究中发现了一个新的细胞内转运蛋白。

关键词： 遗传病家系     基因定位和克隆     基因家族-候选疾病基因克隆     基因组扫描     基因功能研究    

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Overexpression of netrin-1 improves neurological outcomes in mice following transient middle cerebral artery occlusion

null

《医学前沿（英文）》 2011年 第5卷 第1期   页码 86-93 doi: 10.1007/s11684-011-0118-x

摘要：

Netrin-1 (NT-1) is one of the axon-guiding molecules that are critical for neuronal development. Because of its structural homology to the endothelial mitogens, NT-1 may have similar effects on vascular network formation. NT-1 was shown to be able to stimulate the proliferation and migration of human cerebral endothelial cells in vitro and also promote focal neovascularization in adult brain in vivo. In the present study, we reported the delivery of NT-1 using an adeno-associated virus (AAV) vector (AAV-NT-1) into mouse brain followed by transient middle cerebral artery occlusion (tMCAO). We found that AAV vectors did not elicit a detectable inflammatory response, cell loss or neuronal damage after brain transduction. The level of NT-1 was increased in the AAV-NT-1-transduced tMCAO mice compared with the control mice. Furthermore, the neurobehavioral outcomes were significantly improved in AAV-NT-1-transduced mice compared with the control animals (P<0.05) 7 days after tMCAO. Our data suggests that NT-1 plays a neuronal function recovery role in ischemic brain and that NT-1 gene transfer might present a valuable approach to treat brain ischemic disorders.

关键词： adeno-associated virus     angiogenesis     gene transfer     ischemia     middle cerebral artery occlusion     netrin-1    

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水力空化反应器的最新进展——空化机理、反应器设计与应用 Review

Haoxuan Zheng, Ying Zheng, Jesse Zhu

《工程（英文）》 2022年 第19卷 第12期   页码 180-198 doi: 10.1016/j.eng.2022.04.027

摘要：

水力空化因其能效高、操作成本低、能够诱导化学反应并可规模化等优势，被认为是一种很有前途的工艺强化技术。近十年来，人们对水力空化及其主要变量的基本认识取得了一些进展，为水力空化在自由基诱导化学反应过程中的应用提供了基础。在这里，我们对这些研究工作进行了广泛回顾，包括水力空化的基本原理、空化反应器的设计、空化诱导的反应增强及相关的工业应用。比较了两种类型的水力空化反应器，即固定式和旋转式水力空化反应器。讨论了水力空化反应器的设计参数及其在实验室和中试尺度上的反应器性能，并就其最佳操作和几何条件提出了建议。本文首次回顾了目前市场上的商业空化反应器。水力空化的独特特点已广泛应用于各种化学反应，如氧化反应和废水处理，以及如乳化液生成和组分提取等物理过程。文中也详细讨论了自由基和气泡内爆的作用。

关键词： 水动力空化作用     空化核     羟基自由基     固定/旋转水动力空化反应器     反应增强     空化应用    

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标题 作者 时间 类型 操作

Optimized human factor IX expression cassettes for hepatic-directed gene therapy of hemophilia B

null

期刊论文

Construction of lentiviral vector carrying Rab9 gene and its expression in mouse brain

Youguo HAO, Min ZHANG, Jinzhi XU, Bitao BU, Jiajun WEI

期刊论文

Bacteriophages in water pollution control: Advantages and limitations

期刊论文

Assessment of antibiotic resistance genes in dialysis water treatment processes

Xuan Zhu, Chengsong Ye, Yuxin Wang, Lihua Chen, Lin Feng

期刊论文

Matrix attachment regions included in a bicistronic vector enhances and stabilizes follistatin gene expressions

Xiaoming HU,Jing GUO,Chunling BAI,Zhuying WEI,Li GAO,Tingmao HU,Shorgan BOU,Guangpeng LI

期刊论文

Recent progress in hydrodynamic characteristics research and application of annular centrifugal extractors

期刊论文

Invading target cells: multifunctional polymer conjugates as therapeutic nucleic acid carriers

Ulrich L?CHELT, Ernst WAGNER

期刊论文

Identification of pollution sources in rivers using a hydrodynamic diffusion wave model and improved

期刊论文

Numerical analysis of hydrodynamic process of circular-translational-moving polishing (CTMP)

ZHAI Wenjie, LIU Changxiong, LIANG Yingchun

期刊论文

Uncertainty quantification of stability and damage detection parameters of coupled hydrodynamic-ground

Nazim Abdul NARIMAN, Tom LAHMER, Peyman KARAMPOUR

期刊论文

Scenario analysis of water pollution control in the typical peri-urban river using a coupled hydrodynamic-water

Haifeng JIA, Shuo WANG, Mingjie WEI, Yansong ZHANG

期刊论文

Development of a hydrodynamic model and the corresponding virtual software for dual-loop circulating

Shanwei Hu, Xinhua Liu

期刊论文

人类遗传病的家系收集疾病基因定位克隆与疾病基因功能的研究

夏家辉

期刊论文

Overexpression of netrin-1 improves neurological outcomes in mice following transient middle cerebral artery occlusion

null

期刊论文

水力空化反应器的最新进展——空化机理、反应器设计与应用

Haoxuan Zheng, Ying Zheng, Jesse Zhu

期刊论文