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Construction of Hsp90β gene specific silencing plasmid and its transfection efficiency

JI Yewei, NIE Bin, LI Ping, ZHOU Yuanguo, XU Xiaoyu

《医学前沿(英文)》 2007年 第1卷 第3期   页码 253-257 doi: 10.1007/s11684-007-0048-9

摘要: The purpose of this work was to construct the plasmid that could direct the synthesis of siRNA-like transcripts and thus mediate strong and specific repression of human heat shock protein 90β (Hsp90β) gene expression and to compare the transfection efficiency of the plasmids in varying conditions of transfection. Three 64 nt oligos corresponding to different regions of the target gene were chemically synthesized and annealed and were then ligated with pSUPER EGFP1 plasmid and double-digested with HindIII and Π. Recombinant plasmids were transformed into , DH5a, and the colonies were picked and grown in the Amp-agarose. The presence of positive clones was checked by the means of endodigestion and sequencing. Three cell strains, HepG2, Human umbilicus vein endothelium cells (HUVEC) and HeK293, were cultured. Then the plasmids were transfected into the cells at different ratios of plasmid to Lipofectamine. The transfection efficiency was measured by detection of enhanced green fluorescence protein (EGFP). The presence of positive recombinant clones were verified by double-digestion and sequencing. The bases inserted into the plasmids were correct and the positive colonies were named pSuper-Hsp90β1, pSuper-Hsp90β2 and pSuper-Hsp90β3. After optimizing the ratio of plasmid to Lipofectamine, we achieved high transfection efficiency in HeK293 cells. Transfection efficiency was still low in the HepG2 cells. In conclusion, the si-RNA-synthesizing plasmids targeting Hsp90β were constructed and transfected into cells with different transfection efficiency.

关键词: HindIII     transfection efficiency     detection     enhanced     different    

Effects of podocin transfection on CD2AP distribution in HEK293 cells

SHA Yugen, HUANG Songming, ZHANG Aihua, ZHAO Fei, CHEN Ronghua

《医学前沿(英文)》 2008年 第2卷 第1期   页码 35-38 doi: 10.1007/s11684-008-0007-0

摘要: The aim of this paper is to construct a podocin fluorescence expression vector and observe the effects of podocin transfection on CD2AP distribution in HEK293 cells. The pGEMT-easy vector containing the full-length cDNA encoding human podocin was cloned and digested with HI and I. The digested full-length podocin was subcloned into pEGFP-C2. The constructed plasmids were transfected into HEK293 cells and its effects on CD2AP distribution were observed by immunofluorescence. The pEGFP-NPHS2 expression vector was successfully constructed and podocin exclusively located on the HEK293 cell membrane. After podocin transfection, CD2AP redistributed from the perinucleus to the cytoplasm in HEK293 cells. It can be concluded that podocin can recruit CD2AP to redistribute from the perinucleus to the cytoplasm in HEK293 cells.

关键词: pEGFP-NPHS2 expression     transfection     pEGFP-C2     cytoplasm     digested    

Multifunctional peptide conjugated amphiphilic cationic copolymer for enhancing ECs targeting, penetrating and nuclear accumulation

Xinghong Duo, Lingchuang Bai, Jun Wang, Jintang Guo, Xiangkui Ren, Shihai Xia, Wencheng Zhang, Abraham Domb, Yakai Feng

《化学科学与工程前沿(英文)》 2020年 第14卷 第5期   页码 889-901 doi: 10.1007/s11705-020-1919-8

摘要: Gene therapy has drawn great attention in the treatments of many diseases, especially for cardiovascular diseases. However, the development of gene carriers with low cytotoxicity and multitargeting function is still a challenge. Herein, the multitargeting REDV-G-TAT-G-NLS peptide was conjugated to amphiphilic cationic copolymer poly( -caprolactone-co-3(S)-methyl-morpholine-2,5-dione)- -polyethyleneimine (PCLMD- -PEI) via a heterobifunctional orthopyridyl disulfide-poly(ethylene glycol)- -hydroxysuccinimide (OPSS-PEG-NHS) linker to prepare PCLMD- -PEI-PEG-REDV-G-TAT-G-NLS copolymers with the aim to develop the gene carriers with low cytotoxicity and high transfection efficiency. The multitargeting micelles were prepared from PCLMD- -PEI-PEG-REDV-G-TAT-G-NLS copolymers by self-assembly method and used to load pEGFP-ZNF580 plasmids (pDNA) to form gene complexes for enhancing the proliferation and migration of endothelial cells (ECs). The loading pDNA capacity was proved by agarose gel electrophoresis assay. These multitargeting gene complexes exhibited low cytotoxicity by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The high internalization efficiency of these gene complexes was confirmed by flow cytometry. The results of transfection demonstrated that these multitargeting gene complexes possessed relatively high transfection efficiency. The rapid migration of ECs transfected by these gene complexes was verified by wound healing assay. Owing to ECs-targeting ability, cell-penetrating ability and nuclear targeting capacity of REDV-G-TAT-G-NLS peptide, the multitargeting polycationic gene carrier with low cytotoxicity and high transfection efficiency has great potential in gene therapy.

关键词: gene carriers     multitargeting function     ECs     transfection efficiency    

Changes of phenotype and function of human CD4 CD25 T cells induced by transfection of Foxp3

WU Kui, BI Yutian, WANG Yaoli, WANG Changzheng

《医学前沿(英文)》 2008年 第2卷 第4期   页码 366-369 doi: 10.1007/s11684-008-0070-6

摘要: The aim of this paper is to explore the effects of transfection of Foxp3 gene on the phenotype and function of naive CD4 T cells. The pMSCV-Foxp3 retroviral vector encoding Foxp3 gene was transduced into the PT67 packaging cell line. Virus-containing supernatant was applied to differentiate CD4CD25 T cells. The resulting cells were sorted with flow cytometry. The expressions of CD25, CD127, CTLA-4 and the proliferation of transfected T cells were examined. The effect of transfected CD4 T cells on the proliferation and cytokine production of CD4CD25 T cells was examined. Foxp3-gene transfected CD4 T cells could express Foxp3 and transfection of Foxp3 gene up-regulated the expressions of CD25 and CTLA-4, but down-regulated CD127 expression. After transfection, the proliferation of CD4 T cells was eliminated. Transfected T cells inhibited the proliferation of CD4CD25 T cells. CD4CD25 T cells acquired a regulatory phenotype and function after it was transduced with the Foxp3 gene. This suggested a key role of Foxp3 in the generation of CD4CD25 regulatory T cells.

Adenovirus-mediated tissue inhibitor of metalloproteinase-3 gene transfection inhibits rabbit intervertebral

Xudong YU MM, Zengwu SHAO MD, Liming XIONG MD, Weiwei XU MM, Hezhong WANG MM, Huifa XU MM,

《医学前沿(英文)》 2009年 第3卷 第4期   页码 415-420 doi: 10.1007/s11684-009-0072-z

摘要: The aim of this study was to investigate the inhibitory effects of recombinant adenovirus vector carrying tissue inhibitor of metalloproteinase-3 (RAdTIMP-3) against degeneration of rabbit intervertebral disc. Thirty Japanese white rabbits of 4 months old were randomly divided into 5 groups. Mild or moderate rabbit lumbar disc degeneration model was constructed with the controllable axial loading device by imposing 98N pressure at the discs for 2 weeks. Various doses of virus were injected into the degenerated discs as follows: 20μL of normal saline in group 1; 20μL of RAd66 (an empty adenovirus vector, 1.0×10OPU/mL) in group 2; and 20, 10, and 5μL of RAdTIMP-3 (1.0×10OPU/mL) in groups 3, 4, and 5, respectively. Two weeks after the injection, the discs were collected for investigations, including assessment of degeneration degrees according to the Thompson’s grading system, reverse-transcription polymerase chain reaction (RT-PCR) assay for TIMP-3 gene, Safranin O-Fast green staining, and immunohistochemical staining for TIMP-3 and type II collagen. According to Thompson’s criteria, the degeneration of groups 3, 4, and 5, especially group 3, was alleviated as compared with groups 1 and 2. RT-PCR revealed that the expression of TIMP-3 in groups 3, 4, and 5, especially in group 3, was significantly enhanced as compared with group 1 (<0.01). Both Safranin O-Fast green staining and type II collagen staining demonstrated better reserved integrity of disc matrix in groups 3, 4, and 5 than in groups 1 and 2. TIMP-3 staining exhibited an obvious increase of positive-staining rate in groups 3, 4, and 5 as compared with group 1. The positive-staining rate in group 3 (79.42%±1.35%) was about 3times that of group 1 (25.47%±5.46%, <0.01). RAdTIMP-3 can effectively protect the matrix of rabbit intervertebral disc against overloading-induced degeneration in a dose-dependent manner, resulting in the alleviation of disc degeneration.

关键词: tissue inhibitor of metalloproteinase-3     intervertebral disc     rabbit     gene therapy    

The effect of different agricultural management practices on irrigation efficiency, water use efficiency

La ZHUO, Arjen Y. HOEKSTRA

《农业科学与工程前沿(英文)》 2017年 第4卷 第2期   页码 185-194 doi: 10.15302/J-FASE-2017149

摘要: This paper explores the effect of varying agricultural management practices on different water efficiency indicators: irrigation efficiency (IE), crop water use efficiency (WUE), and green and blue water footprint (WF). We take winter wheat in an experimental field in Northern China as a case study and consider a dry, average and wet year. We conducted 24 modeling experiments with the AquaCrop model, for all possible combinations of four irrigation techniques, two irrigation strategies and three mulching methods. Results show that deficit irrigation most effectively improved blue water use, by increasing IE (by 5%) and reducing blue WF (by 38%), however with an average 9% yield reduction. Organic or synthetic mulching practices improved WUE (by 4% and 10%, respectively) and reduced blue WF (by 8% and 17%, respectively), with the same yield level. Drip and subsurface drip irrigation improved IE and WUE, but drip irrigation had a relatively large blue WF. Improvements in one water efficiency indicator may cause a decline in another. In particular, WUE can be improved by more irrigation at the cost of the blue WF. Furthermore, increasing IE, for instance by installing drip irrigation, does not necessarily reduce the blue WF.

关键词: field management     irrigation efficiency     water footprint     water productivity     water use efficiency    

Gene delivery into isolated

Nan Zheng, Ziyuan Song, Yang Liu, Lichen Yin, Jianjun Cheng

《化学科学与工程前沿(英文)》 2017年 第11卷 第4期   页码 521-528 doi: 10.1007/s11705-017-1612-8

摘要: The application of gene delivery materials has been mainly focused on mammalian cells while rarely extended to plant engineering. Cationic polymers and lipids have been widely utilized to efficiently deliver DNA and siRNA into mammalian cells. However, their application in plant cells is limited due to the different membrane structures and the presence of plant cell walls. In this study, we developed the cationic, -helical polypeptide that can effectively deliver DNA into both isolated protoplasts and intact leaves. The PPABLG was able to condense DNA to form nanocomplexes, and they exhibited significantly improved transfection efficiencies compared with commercial transfection reagent Lipofectamine 2000 and classical cell penetrating peptides such as poly(L-lysine), HIV-TAT, arginine9, and poly(L-arginine). This study therefore widens the utilities of helical polypeptide as a unique category of gene delivery materials, and may find their promising applications toward plant gene delivery.

关键词: α-helical polypeptide     plant gene delivery     protoplast     intact leaves     transfection    

Evaluating R&D efficiency of China’s listed lithium battery enterprises

《工程管理前沿(英文)》   页码 473-485 doi: 10.1007/s42524-022-0213-5

摘要: Promoting the growth of the lithium battery sector has been a critical aspect of China’s energy policy in terms of achieving carbon neutrality. However, despite significant support on research and development (R&D) investments that have resulted in increasing size, the sector seems to be falling behind in technological areas. To guide future policies and understand proper ways of promoting R&D efficiency, we looked into the lithium battery industry of China. Specifically, data envelopment analysis (DEA) was used as the primary approach based on evidence from 22 listed lithium battery enterprises. The performance of the five leading players was compared with that of the industry as a whole. Results revealed little indication of a meaningful improvement in R&D efficiency throughout our sample from 2010 to 2019. However, during this period, a significant increase in R&D expenditure was witnessed. This finding was supported, as the results showed that the average technical efficiency of the 22 enterprises was 0.442, whereas the average pure technical efficiency was at 0.503, thus suggesting that they were suffering from decreasing returns to scale (DRS). In contrast, the performance of the five leading players seemed superior because their average efficiency scores were higher than the industry’s average. Moreover, they were experiencing increasing scale efficiency (IRS). We draw on these findings to suggest to policymakers that supporting technologically intensive sectors should be more than simply increasing investment scale; rather, it should also encompass assisting businesses in developing efficient managerial processes for R&D.

关键词: Data Envelopment Analysis     R&D investment efficiency     China’s listed lithium battery enterprises     technical efficiency     pure technical efficiency     scale efficiency    

Improving the efficiency and effectiveness of global phosphorus use: focus on root and rhizosphere levels

null

《农业科学与工程前沿(英文)》 2019年 第6卷 第4期   页码 357-365 doi: 10.15302/J-FASE-2019275

摘要:

Phosphorus (P) is essential for life and for efficient crop production, but global P use with limited recycling is inefficient in several sectors, including agronomy. Unfortunately, plant physiologists, agronomists, farmers and end users employ different measures for P use efficiency (PUE), which often masks their values at different scales. The term P use effectiveness, which also considers energetic and sustainability measures in addition to P balances, is also a valuable concept. Major physiological and genetic factors for plant P uptake and utilization have been identified, but there has been limited success in genetically improving PUE of modern crop cultivars. In maize, studies on root architectural and morphological traits appear promising. Rhizosphere processes assist in mobilizing and capturing sparingly soluble phosphate from rock phosphate. Combinations of phosphate-solubilizing microorganisms with ammonium-based nitrogen fertilizer, as well as strategies of fertilizer placement near the roots of target crops, can moderately enhance PUE. The desired concentration of P in the products differs, depending on the final use of the crop products as feed, food or for energy conversion, which should be considered during crop production.

关键词: acquisition efficiency     plant growth promoting rhizobacteria     phosphate     use efficiency     utilization efficiency    

Optimization of power and efficiency for an irreversible Diesel heat engine

Shiyan ZHENG, Guoxing LIN

《能源前沿(英文)》 2010年 第4卷 第4期   页码 560-565 doi: 10.1007/s11708-010-0018-9

摘要: A cyclic model of an irreversible Diesel heat engine is presented, in which the heat loss between the working fluid and the ambient during combustion, the irreversibility inside the cyclic working fluid resulting from friction, eddies flow, and other irreversible effects are taken into account. By using the thermodynamic analysis and optimal control theory methods, the analytical expressions of power output and efficiency of the Diesel heat engine are derived. Variations of the main performance parameters with the pressure ratio of the cycle are analyzed and calculated. The optimum operating region of the heat engine is determined. Moreover, the optimum criterion of some important parameters, such as the power output, efficiency, pressure ratio, and temperatures of the working fluid at the related state points are illustrated and discussed. The conclusions obtained in the present paper may provide some theoretical guidance for the optimal parameter design of a class of internal-combustion engines.

关键词: Diesel heat engine     irreversibility     power output     efficiency     parameter optimization    

Methods and applications of DEA cross-efficiency: Review and future perspectives

Jie WU, Jiasen SUN, Liang LIANG

《工程管理前沿(英文)》 2021年 第8卷 第2期   页码 199-211 doi: 10.1007/s42524-020-0133-1

摘要: The field of engineering management usually involves evaluation issues, such as program selection, team performance evaluation, technology selection, and supplier evaluation. The traditional self-evaluation data envelopment analysis (DEA) method usually exaggerates the effects of several inputs or outputs of the evaluated decision-making unit (DMU), resulting in unrealistic results. To address this problem, scholars have proposed the cross-efficiency evaluation (CREE) method. Compared with the DEA method, CREE can rank DMUs more completely by using reasonable weights. With the extensive application of this technique, several problems, such as non-unique weights and non-Pareto optimal results, have arisen in CREE methods. Therefore, the improvement of CREE has attracted the attention of many scholars. This paper reviews the theory and applications of CREE, including the non-uniqueness problem, the aggregation of cross-efficiency data, and applications in engineering management. It also discusses the directions for future research on CREE.

关键词: cross-efficiency evaluation     efficiency     secondary goal model     aggregation     review    

STRATEGIES FOR IMPROVING WATER USE EFFICIENCY IN DRYLAND AGROECOSYSTEMS

《农业科学与工程前沿(英文)》 2021年 第8卷 第4期   页码 599-602 doi: 10.15302/J-FASE -2021409

Effects of dwarfing genes on water use efficiency of bread wheat

Jiakun YAN, Suiqi ZHANG

《农业科学与工程前沿(英文)》 2017年 第4卷 第2期   页码 126-134 doi: 10.15302/J-FASE-2017134

摘要: Climate change has increased the risk of drought, which significantly limits plant productivity. Various ways of increasing water availability and sustaining growth of crop plants in drought-prone environments are available. Genetic advances in grain yields under rainfed conditions have been achieved with the introduction of dwarfing genes. A thorough understanding of the effects of different dwarfing genes on root growth, coleoptile length, grain yields and water using efficiency (WUE) will provide opportunities to select appropriate genes for breeding high WUE and grain yield cultivars. This review focuses on the mechanism involved in genes that reduce plant height and affect root and coleoptile length, their consequent effects on grain yields and WUE, and suggests that for rainfed and irrigation-limited environments, combining GAR and GAI dwarfing genes in breeding may help boost WUE and yields, and more materials from different parental sources should be collected to assess opportunities for potential comprehensive application of specific genes.

关键词: coleoptile length     wheat     dwarfing genes     grain yield     root     water use efficiency    

neutrality potential of wastewater treatment plants: A novel evaluation framework integrating energy efficiency

《环境科学与工程前沿(英文)》 2022年 第16卷 第9期 doi: 10.1007/s11783-022-1549-0

摘要:

• Framework of indicators was established based on energy efficiency and recovery.

关键词: Wastewater treatment plants     Energy neutrality potential     Energy efficiency     Energy recovery     Evaluation framework    

Approaches to achieve high grain yield and high resource use efficiency in rice

Jianchang YANG

《农业科学与工程前沿(英文)》 2015年 第2卷 第2期   页码 115-123 doi: 10.15302/J-FASE-2015055

摘要: This article discusses approaches to simultaneously increase grain yield and resource use efficiency in rice. Breeding nitrogen efficient cultivars without sacrificing rice yield potential, improving grain fill in later-flowering inferior spikelets and enhancing harvest index are three important approaches to achieving the dual goal of high grain yield and high resource use efficiency. Deeper root distribution and higher leaf photosynthetic N use efficiency at lower N rates could be used as selection criteria to develop N-efficient cultivars. Enhancing sink activity through increasing sugar-spikelet ratio at the heading time and enhancing the conversion efficiency from sucrose to starch though increasing the ratio of abscisic acid to ethylene in grains during grain fill could effectively improve grain fill in inferior spikelets. Several practices, such as post-anthesis controlled soil drying, an alternate wetting and moderate soil drying regime during the whole growing season, and non-flooded straw mulching cultivation, could substantially increase grain yield and water use efficiency, mainly via enhanced remobilization of stored carbon from vegetative tissues to grains and improved harvest index. Further research is needed to understand synergistic interaction between water and N on crop and soil and the mechanism underlying high resource use efficiency in high-yielding rice.

关键词: rice     nitrogen-efficient cultivar     grain fill     harvest index     nitrogen use efficiency     water use efficiency    

标题 作者 时间 类型 操作

Construction of Hsp90β gene specific silencing plasmid and its transfection efficiency

JI Yewei, NIE Bin, LI Ping, ZHOU Yuanguo, XU Xiaoyu

期刊论文

Effects of podocin transfection on CD2AP distribution in HEK293 cells

SHA Yugen, HUANG Songming, ZHANG Aihua, ZHAO Fei, CHEN Ronghua

期刊论文

Multifunctional peptide conjugated amphiphilic cationic copolymer for enhancing ECs targeting, penetrating and nuclear accumulation

Xinghong Duo, Lingchuang Bai, Jun Wang, Jintang Guo, Xiangkui Ren, Shihai Xia, Wencheng Zhang, Abraham Domb, Yakai Feng

期刊论文

Changes of phenotype and function of human CD4 CD25 T cells induced by transfection of Foxp3

WU Kui, BI Yutian, WANG Yaoli, WANG Changzheng

期刊论文

Adenovirus-mediated tissue inhibitor of metalloproteinase-3 gene transfection inhibits rabbit intervertebral

Xudong YU MM, Zengwu SHAO MD, Liming XIONG MD, Weiwei XU MM, Hezhong WANG MM, Huifa XU MM,

期刊论文

The effect of different agricultural management practices on irrigation efficiency, water use efficiency

La ZHUO, Arjen Y. HOEKSTRA

期刊论文

Gene delivery into isolated

Nan Zheng, Ziyuan Song, Yang Liu, Lichen Yin, Jianjun Cheng

期刊论文

Evaluating R&D efficiency of China’s listed lithium battery enterprises

期刊论文

Improving the efficiency and effectiveness of global phosphorus use: focus on root and rhizosphere levels

null

期刊论文

Optimization of power and efficiency for an irreversible Diesel heat engine

Shiyan ZHENG, Guoxing LIN

期刊论文

Methods and applications of DEA cross-efficiency: Review and future perspectives

Jie WU, Jiasen SUN, Liang LIANG

期刊论文

STRATEGIES FOR IMPROVING WATER USE EFFICIENCY IN DRYLAND AGROECOSYSTEMS

期刊论文

Effects of dwarfing genes on water use efficiency of bread wheat

Jiakun YAN, Suiqi ZHANG

期刊论文

neutrality potential of wastewater treatment plants: A novel evaluation framework integrating energy efficiency

期刊论文

Approaches to achieve high grain yield and high resource use efficiency in rice

Jianchang YANG

期刊论文