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Expression of recombinant human butyrylcholinesterase in the milk of transgenic mice

Dan LU,Shengzhe SHANG,Shen LIU,Ying WU,Fangfang WU,Tan TAN,Qiuyan LI,Yunping DAI,Xiaoxiang HU,Yaofeng ZHAO,Ning LI

《农业科学与工程前沿（英文）》 2014年 第1卷 第3期   页码 179-184 doi: 10.15302/J-FASE-2014020

摘要： Butyrylcholinesterase (BCHE) is a natural bioscavenger that protects humans against organophosphate toxicity. Due to the limited yield of human BCHE (hBCHE) when purifying from human plasma, it is necessary to find an alternative method to produce this protein. One potential method is to produce transgenic livestock that make modified milk containing high concentration of hBCHE. In this study, we cloned the gene into a human lactoferrin (hLF) bacterial artificial chromosome (BAC) construct to make a hLF-hBCHE BAC construct. Subsequently, we injected the BAC construct into pronuclei of mouse fertilized embryos and generated transgenic mice. Expression analysis showed that recombinant hBCHE (rhBCHE) was expressed efficiently in the mammary gland of the transgenic mice and the concentration of rhBCHE in the milk of individual mice ranged from 76±12 to 159±28 mg·L . Protein function tests showed that rhBCHE has the same enzymatic activity as the native hBCHE. Our results pave the way for making transgenic livestock to produce large quantities of rhBCHE.

关键词： recombinant human butyrylcholinesterase (rhBCHE)     human lactoferrin bacterial artificial chromosome (hLF BAC)     transgenic mice     milk    

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A bacterial artificial chromosome-based physical map of

Yuhua FU,Zhiqiang XIA,Shujuan WANG,Xin CHEN,Cheng LU,Mingcheng LUO,Hongbin ZHANG,Wenquan WANG

《农业科学与工程前沿（英文）》 2016年 第3卷 第4期   页码 321-329 doi: 10.15302/J-FASE-2016124

摘要： Cassava ( ) is known as the third most important food crop in the tropics and also used for industrial feedstock for biofuels. Two new bacterial artificial chromosome (BAC) libraries were constructed for W14 ( ssp. ), a wild ancestor of domesticated cassava. The libraries were constructed with RI and dIII insertion vectors, respectively. The RI library has 29952 clones with an average insert size of 115 kb, while the dIII library consists of 29952 clones with an average insert of 129 kb. The combined libraries contain a total of 59904 clones with an average insert size of 125 kb, representing approximately 10 × haploid genome equivalents. A total of 29952 clones were fingerprinted and resulted in a cassava physical map composed of 2485 contigs with an average physical length of 336 kb and 2909 singletons, representing approximately 762 Mb of the cassava genome. 5000 clones located at the ends of BAC contigs were selected and sequenced. A total of 6077 SNPs and 231 indels were identified, that covered 459 gene sequences, of which 6 genes were associated with starch and sucrose metabolism. This BAC-based physical map provides valuable tools to understand the genetics and evolution of cassava.

关键词： cassava     BAC library     physical map    

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Development of pretreatment protocol for DNA extraction from biofilm attached to biologic activated carbon (BAC

Shuting ZHANG, Bo WEI, Xin YU, Bing LIU, Zhuoying WU, Li GU

《环境科学与工程前沿（英文）》 2010年 第4卷 第4期   页码 459-465 doi: 10.1007/s11783-010-0249-3

摘要： The biologic activated carbon (BAC) process is widely used in drinking water treatments. A comprehensive molecular analysis of the microbial community structure provides very helpful data to improve the reactor performance. However, the bottleneck of deoxyribonucleic acid (DNA) extraction from BAC attached biofilm has to be solved since the conventional procedure was unsuccessful due to firm biomass attachment and adsorption capacity of the BAC granules. In this study, five pretreatments were compared, and adding skim milk followed by ultrasonic vibration was proven to be the optimal choice. This protocol was further tested using the vertical BAC samples from the full-scale biofilter of Pinghu Water Plant. The results showed the DNA yielded a range of 40 μg·g BAC (dry weight) to over 100 μg·g BAC (dry weight), which were consistent with the biomass distribution. All results suggested that the final protocol could produce qualified genomic DNA as a template from the BAC filter for downstream molecular biology researches.

关键词： bacterial DNA extraction     biological activated carbon (BAC)     biofilm     water treatment     pretreatment protocol    

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Construction of a universal recombinant expression vector that regulates the expression of human lysozyme

Shen LIU, Shengzhe SHANG, Xuezhen YANG, Huihua ZHANG, Dan LU, Ning LI

《农业科学与工程前沿（英文）》 2018年 第5卷 第3期   页码 382-389 doi: 10.15302/J-FASE-2018211

摘要：

The mammary gland provides a novel method for producing recombinant proteins in milk of transgenic animals. A key component in the technology is the construction of an efficient milk expression vector. Here, we established a simple method to construct a milk expression vector, by a combination of homologous recombination and digestion-ligation. Our methodology is expected to have the advantages of both plasmid and bacterial artificial chromosome (BAC) vectors. The BAC of mouse whey acidic protein gene (mWAP) was modified twice by homologous recombination to produce a universal expression vector, and the human lysozyme gene (hLZ) was then inserted into the vector by a digestion-ligation method. The final vector containing the 8.5 kb mWAP 5′ promoter, 4.8 kb hLZ genomic DNA, and 8.0 kb mWAP 3′ genomic DNA was microinjected into pronuclei of fertilized mouse embryos, to successfully generate two transgenic mouse lines that expressed recombinant human lysozyme (rhLZ) in milk. The highest expression level of rhLZ was 0.45 g·L−1, and rhLZ exhibited the same antibacterial activity as native hLZ. Our results have provided a simple approach to construct a universal milk expression vector, and demonstrated that the resulting vector regulates the expression of hLZ in milk.

关键词： BAC recombinant methods     gene expression     human lysozyme     transgenic mice     milk expression vector    

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Impact of preozonation on the bioactivity and biodiversity of subsequent biofilters under low temperature conditions—A pilot study

Jiaxuan YANG, Jun MA, Dan SONG, Xuedong ZHAI, Xiujuan KONG

《环境科学与工程前沿（英文）》 2016年 第10卷 第4期 doi: 10.1007/s11783-016-0844-z

摘要： The bioactivity was enhanced by preozonation under low temperature conditions. Higher level of BDOC/AOC and DO may enhance the nitrifying performance. High level of biodiversity and bioactivity may help maintain the stability of filters. Water quality and DO could selectively enhance different microbial communities. The combination of preozonation and subsequent biological granular activated carbon (O /BAC) filtration is well known as a promising method for the removal of many pollutants. Temperature and nutrients are the dominant factors in external conditions to influence the biological communities. To explore the influence of preozonation under low temperature, the factors such as dissolved oxygen (DO), dissolved organic carbon (DOC) and NH -N were analyzed from the sampling ports every week; triphenyl tetrazolium chloride-dehydrogenase activity (TTC-DHA) and the nitrifying activity were detected along the bed height of biofilter at four levels (10, 40, 70 and 110 cm) on the 90th, 110th, and 130th day; microbial community, based on 16S rRNA gene-denaturing gradient gel electrophoresis (DGGE), was monitored on the 130th day of the operation. The observed microbial property showed that preozonation had a positive influence on bioactivity, biomass and nitrifying activity. Community analysis showed no significant difference on the biodiversity of nitrifying bacteria between the parallel filters in the inlet end based on the method employed. This result showed that biofilters’ performance is not correlated well with microbial biodiversity. The elevated functionality in O /BAC filters can be a result of increased microbial activity, which was promoted by preozonation.

关键词： Preozonation     BAC filtration     High ammonia level     Bacterial activity     Microbial community structure     Pilot scale study    

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Transport of bacterial cell (

Wei Fan, Qi Li, Mingxin Huo, Xiaoyu Wang, Shanshan Lin

《环境科学与工程前沿（英文）》 2020年 第14卷 第4期 doi: 10.1007/s11783-020-1242-0

摘要： Abstract • The recharge pond dwelling process induced changes in cell properties. • Cell properties and solution chemistry exerted confounding effect on cell transport. • E. coli cells within different recharge water displayed different spreading risks. Commonly used recharge water resources for artificial groundwater recharge (AGR) such as secondary effluent (SE), river water and rainfall, are all oligotrophic, with low ionic strengths and different cationic compositions. The dwelling process in recharge pond imposed physiologic stress on Escherichia coli (E. coli) cells, in all three types of investigated recharge water resources and the cultivation of E. coli under varying recharge water conditions, induced changes in cell properties. During adaptation to the recharge water environment, the zeta potential of cells became more negative, the hydrodynamic diameters, extracellular polymeric substances content and surface hydrophobicity decreased, while the cellular outer membrane protein profiles became more diverse. The mobility of cells altered in accordance with changes in these cell properties. The E. coli cells in rainfall recharge water displayed the highest mobility (least retention), followed by cells in river water and finally SE cells, which had the lowest mobility. Simulated column experiments and quantitative modeling confirmed that the cellular properties, driven by the physiologic state of cells in different recharge water matrices and the solution chemistry, exerted synergistic effects on cell transport behavior. The findings of this study contribute to an improved understanding of E. coli transport in actual AGR scenarios and prediction of spreading risk in different recharge water sources.

关键词： Artificial groundwater recharge     E. coli     Transport     Simulated column experiments     Modeling    

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人工智能与统计分析 Perspective

Bin YU, Karl KUMBIER

《信息与电子工程前沿（英文）》 2018年 第19卷 第1期   页码 6-9 doi: 10.1631/FITEE.1700813

摘要： 人工智能（artificial intelligence, AI）本质上是由数据驱动的。在其通过人机协作完成数据生成、算法开发与结果评估的任务中，需要应用许多统计概念。

关键词： 人工智能；统计；人机协作    

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The imperative need to develop guidelines to manage human versus machine intelligence

Donald KENNEDY, Simon P. PHILBIN

《工程管理前沿（英文）》 2018年 第5卷 第2期   页码 182-194 doi: 10.15302/J-FEM-2018085

摘要： Machine intelligence is increasingly entering roles that were until recently dominated by human intelligence. As humans now depend upon machines to perform various tasks and operations, there appears to be a risk that humans are losing the necessary skills associated with producing competitively advantageous decisions. Therefore, this research explores the emerging area of human versus machine decision-making. An illustrative engineering case involving a joint machine and human decision-making system is presented to demonstrate how the outcome was not satisfactorily managed for all the parties involved. This is accompanied by a novel framework and research agenda to highlight areas of concern for engineering managers. We offer that the speed at which new human-machine interactions are being encountered by engineering managers suggests that an urgent need exists to develop a robust body of knowledge to provide sound guidance to situations where human and machine decisions conflict. Human-machine systems are becoming pervasive yet this research has revealed that current technological approaches are not adequate. The engineering insights and multi-criteria decision-making tool from this research significantly advance our understanding of this important area.

关键词： human intelligence & machine intelligence     HI-MI     decision-making     artificial intelligence    

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Generation of CRISPR/Cas9-mediated lactoferrin-targeted mice by pronuclear injection of plasmid pX330

Mengxu GE,Fei LIU,Fei CHANG,Zhaolin SUN,Jing FEI,Ying GUO,Yunping DAI,Zhengquan YU,Yaofeng ZHAO,Ning LI,Qingyong MENG

《农业科学与工程前沿（英文）》 2015年 第2卷 第3期   页码 242-248 doi: 10.15302/J-FASE-2015059

摘要： Lactoferrin is a member of the transferrin family of multifunctional iron binding glycoproteins. While numerous physiological functions have been described for lactoferrin, the mechanisms underlying these functions are not clear. To further study the functions and mechanisms of lactoferrin, we modified the lactoferrin promoter of mice using the CRISPR/Cas9 system to reduce or eliminate lactoferrin expression. Seven mice with lactoferrin promoter mutations were obtained with an efficiency of 24% (7/29) by injecting the plasmid pX330, expressing a small guide RNA and human codon-optimized SpCas9, into fertilized eggs of mice. Plasmid integration and off-targeting of pX330 were not detected. These results confirmed that pronuclear injection of a circular plasmid is a feasible and efficient method for targeted mutagenesis in mice.

关键词： lactoferrin     promoter     CRISPR/Cas9     plasmid pX330    

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智能源于人、拓于工——人工智能发展的一点思考

蒋昌俊,王俊丽

《中国工程科学》 2018年 第20卷 第6期   页码 93-100 doi: 10.15302/J-SSCAE-2018.06.015

摘要：

人工智能（AI）旨在模拟人脑中信息存储和处理机制等智能行为，使机器具有一定程度的智能水平。随着互联网、大数据、云计算和深度学习等新一代信息技术的飞速发展，目前AI领域的研究和应用已经取得重要进展。本文将深入分析与AI密切相关的计算机科学、控制科学、类脑智能、人脑智能等学科或领域之间的交融与历史演进；指出神经科学、脑科学与认知科学中有关脑的结构与功能机制的研究成果，为构建智能计算模型提供了重要的启发，并从逻辑模型及系统、神经元及网络模型、视觉神经分层机制等方面，分别阐述智能的驱动与发展；最后从互联网的计算理论、AI的演算和计算的融合、类脑智能的模型和机理、AI对神经科学的推动作用、反馈计算的算法设计与控制系统的能级五个方面，对AI的发展趋势进行了展望。

关键词： 人工智能     人脑智能     类脑智能     智能发展     学科演进    

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Philadelphia chromosome-positive acute myeloid leukemia with masses and osteolytic lesions: finding of

null

《医学前沿（英文）》 2017年 第11卷 第3期   页码 440-444 doi: 10.1007/s11684-017-0523-x

摘要：

Philadelphia chromosome-positive acute myeloid leukemia is controversial and difficult to distinguish from the blast phase of chronic myeloid leukemia. As a myeloid neoplasm, rare cases of this leukemia manifest multiple soft-tissue tumors or bone lytic lesions. In this paper, we describe a 49-year-old male patient who had an abrupt onset with sharp chest pain, fever, fatigue, emaciation, and splenomegaly. 18F-fluoro-deoxy-glucose positron emission tomography/computed tomography (18F-FDG PET/CT) result showed diffuse and uneven hypermetabolic lesions in the bone marrow with peripheral bone marrow expansion, multiple soft tissue neoplasms with high 18F-FDG uptake, and lytic bone lesions. Bone marrow smear and biopsy detected aberrant blast cells expressing myeloid rather than lymphoid immunophenotype marker. For the existence of Philadelphia chromosome and BCR-ABL1 fusion gene together with complex chromosome abnormalities, a diagnosis of Philadelphia-positive acute myeloid leukemia was made, although the type (de novo or blast crisis) remained unclear.

关键词： Philadelphia chromosome     acute myeloid leukemia     mass     osteolysis     positron emission tomography    

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prognostic factors of imatinib plus CALLG2008 protocol in adult patients with newly diagnosed Philadelphia chromosome-positive

null

《医学前沿（英文）》 2017年 第11卷 第2期   页码 229-238 doi: 10.1007/s11684-017-0506-y

摘要：

A CALLG2008 protocol was developed by the Chinese Acute Lymphoblastic Leukemia Cooperative Group for adult acute lymphoblastic leukemia (ALL). We retrospectively analyzed 153 newly diagnosed adult patients with Philadelphia chromosome (Ph)-positive ALL enrolled into imatinib (400 mg/d) plus CALLG2008 regimen between 2009 and 2015. The median age was 40 years (range, 18–68 years), with 81 (52.3%) males. The overall hematologic complete remission (CR) rate was 96.7% after induction. With a median follow-up of 24.2 months, the estimated 3-year overall survival (OS) and event-free survival (EFS) rates were 49.5% (95% confidence interval (CI): 38.5%–59.5%) and 49.2% (95% CI: 38.3%–59.2%), respectively. Fifty-eight (36 with haploidentical donor) patients underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) in first CR. Among the patients in CR1 after induction, both the 3-year OS and EFS were significantly better in the allo-HSCT group than in the without allo-HSCT group (73.2%, 95% CI: 58.3%–83.5% vs. 22.2%, 95% CI: 8.7%–39.6% and 66.5%, 95% CI: 50.7%–78.2% vs. 16.1%, 95% CI: 5.1%–32.7%, respectively). Multivariate analysis showed that allo-HSCT and achievement of major molecular response were associated with favorable OS or EFS independently. Interestingly, in the allo-HSCT cohort, the donor type (haploidentical versus matched donors) had no significant impact on EFS or OS. All these results suggested that imatinib plus CALLG2008 was an effective protocol for Ph-positive ALL. Haploidentical donors can also be a reasonable alternative expedient donor pool.

关键词： Philadelphia chromosome     acute lymphoblastic leukemia     imatinib     CALLG2008    

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人工智能走向2.0

潘云鹤

《工程（英文）》 2016年 第2卷 第4期   页码 409-413 doi: 10.1016/J.ENG.2016.04.018

摘要：

随着互联网的普及、传感网的渗透、大数据的涌现、信息社区的崛起，以及数据和信息在人类社会、物理空间和信息空间之间的交叉融合与相互作用，当今人工智能(AI) 发展所处信息环境和数据基础已经发生了深刻变化，人工智能的目标和理念正面临重要调整，人工智能的科学基础和实现载体也面临新的突破，人工智能正进入一个新的阶段。这个源于传统而又与之不同的人工智能新阶段被称为人工智能2.0(AI 2.0)。本文从人工智能60 年的发展历史出发，通过分析促成人工智能2.0形成的外部环境与目标的转变，分析技术萌芽，提出了人工智能2.0 的核心理念，并结合中国发展的社会需求与信息环境特色，给出了发展人工智能2.0 的建议。

关键词： 人工智能2.0     大数据     群体智能     跨媒体     人机混合智能     无人智能系统    

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The predictive value of chromosome 8p deletion for metastasis of hepatocellular carcinoma: a summary

QIN Lunxiu, TANG Zhaoyou, GUAN Xinyuan, YE Qinghai, JIA Huliang, REN Ning

《医学前沿（英文）》 2008年 第2卷 第3期   页码 211-215 doi: 10.1007/s11684-008-0041-y

摘要： Hepatocellular carcinoma (HCC) represents an extremely poor prognostic cancer, which is mainly due to the high frequency of metastasis/recurrence after surgical operation. Exploring the molecular mechanisms involved in HCC metastasis could be helpful in the prediction and early diagnosis of HCC recurrence and could also provide new therapeutic targets for HCC metastasis. In the recent decade, we analyzed the genomic aberrations of the clinical specimens, as well as the metastatic models and cell lines of human HCC to identify the genetic markers related to HCC metastasis and to verify their clinical values in the prediction and control of metastasis of HCC. Using the comparative genomic hybridization (CGH) technique, we compared the differences of chromosomal aberrations between primary HCC tumors and their matched metastatic lesions, and found that chromosome 8p deletions might contribute to HCC metastasis. This novel finding was further confirmed by comparison between nude mice models of HCC with different metastatic potentials. By the more sensitive genome-wide microsatellite analysis, 8p deletion was defined to 8p23.3 and 8p11.2, which are two likely regions harboring metastasis-related genes of HCC. Using ‘8p-specific’ microarrays, two novel metastatic suppressors ( and ) were identified, and were proven to suppress invasion and metastasis of HCC. Clinical studies indicate that 8p deletion detected in HCC or circulating plasma DNA of patients is a useful predictor for metastatic recurrence and prognosis, even for patients with early stage HCC. These novel findings are regarded as important advances in the study of the molecular mechanisms of HCC metastasis, which provide not only a holistic view on the molecular cytogenetic bases of HCC metastasis, but also candidate regions for further study to identify metastatic suppressor genes.

关键词： sensitive genome-wide     prediction     genome-wide microsatellite     frequency     Hepatocellular carcinoma    

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Antibiotic resistome mostly relates to bacterial taxonomy along a suburban transmission chain

《环境科学与工程前沿（英文）》 2022年 第16卷 第3期 doi: 10.1007/s11783-021-1466-7

摘要：

• The α-diversities of resistome were lower in manure and compost than in soils.

关键词： Antibiotic resistance genes     Resistome     Bacterial taxonomy     Transmission chain    

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标题 作者 时间 类型 操作

Expression of recombinant human butyrylcholinesterase in the milk of transgenic mice

Dan LU,Shengzhe SHANG,Shen LIU,Ying WU,Fangfang WU,Tan TAN,Qiuyan LI,Yunping DAI,Xiaoxiang HU,Yaofeng ZHAO,Ning LI

期刊论文

A bacterial artificial chromosome-based physical map of

Yuhua FU,Zhiqiang XIA,Shujuan WANG,Xin CHEN,Cheng LU,Mingcheng LUO,Hongbin ZHANG,Wenquan WANG

期刊论文

Development of pretreatment protocol for DNA extraction from biofilm attached to biologic activated carbon (BAC

Shuting ZHANG, Bo WEI, Xin YU, Bing LIU, Zhuoying WU, Li GU

期刊论文

Construction of a universal recombinant expression vector that regulates the expression of human lysozyme

Shen LIU, Shengzhe SHANG, Xuezhen YANG, Huihua ZHANG, Dan LU, Ning LI

期刊论文

Impact of preozonation on the bioactivity and biodiversity of subsequent biofilters under low temperature conditions—A pilot study

Jiaxuan YANG, Jun MA, Dan SONG, Xuedong ZHAI, Xiujuan KONG

期刊论文

Transport of bacterial cell (

Wei Fan, Qi Li, Mingxin Huo, Xiaoyu Wang, Shanshan Lin

期刊论文

人工智能与统计分析

Bin YU, Karl KUMBIER

期刊论文

The imperative need to develop guidelines to manage human versus machine intelligence

Donald KENNEDY, Simon P. PHILBIN

期刊论文

Generation of CRISPR/Cas9-mediated lactoferrin-targeted mice by pronuclear injection of plasmid pX330

Mengxu GE,Fei LIU,Fei CHANG,Zhaolin SUN,Jing FEI,Ying GUO,Yunping DAI,Zhengquan YU,Yaofeng ZHAO,Ning LI,Qingyong MENG

期刊论文

智能源于人、拓于工——人工智能发展的一点思考

蒋昌俊,王俊丽

期刊论文

Philadelphia chromosome-positive acute myeloid leukemia with masses and osteolytic lesions: finding of

null

期刊论文

prognostic factors of imatinib plus CALLG2008 protocol in adult patients with newly diagnosed Philadelphia chromosome-positive

null

期刊论文

人工智能走向2.0

潘云鹤

期刊论文

The predictive value of chromosome 8p deletion for metastasis of hepatocellular carcinoma: a summary

QIN Lunxiu, TANG Zhaoyou, GUAN Xinyuan, YE Qinghai, JIA Huliang, REN Ning

期刊论文

Antibiotic resistome mostly relates to bacterial taxonomy along a suburban transmission chain

期刊论文