Wang Sheng

Tobacco Research Institute

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Co-Authors

Xiangzhen Ding

Ningxia University

Fuwei Xie

Tobacco Research Institute

Xie Fuwei

C3J Therapeutics (United States)

Jie Dong

Shanghai Jiao Tong University

Zhao Ge

Huazhong University of Science and Technology

Yigang Tong

Beijing University of Chemical Technology

Xiaoping An

Beijing University of Chemical Technology

Jiankui Chen

Huazhong University of Science and Technology

Bing Wang

Shandong Institute for Product Quality Inspection

MU Hong-zhen

Cooperative Institutions

Chinese Academy of Sciences

Tobacco Research Institute

Chinese Academy of Medical Sciences & Peking Union Medical College

Ministry of Agriculture and Rural Affairs

Zhejiang University

Zhengzhou University

Wuhan University

Peking University

Guangxi Veterinary Research Institute

Ministry of Education of the People's Republic of China

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A method for rapid separation and determination of aconitine in plasma by reversed-phase high performance liquid chromatography

Journal of Forensic Medicine (2005)

Wang Sheng

Objective For the purpose of more accurate and rapid analysis of aconitine in plasma, a method by reversed-phase high performance liquid chromatography (HPLC) was developed. Methods Standard aconitine was added to the blank plasma after the plasma sample was pretreated. The sample pretreatment method, linear range, the precision, and the recoveries for the plasma were investigated by using of human plasma spiked with standard aconitine. Results The linear range was 0.1~5.0 μgmL with (γ=0.9998). The limit of detection was 0.1μgmL. The intra and inter-day precisions of assay for aconi-tine were less than 3.7% and 4.5% (n=5) in blood respectively. The recovery for aconitine was more than (97.0±4.1)% in plasma. The HPLC method was thereafter used to determine the concentration of aconitine in some forensic cases. Conclusion The HPLC method for quantitative analysis of aconitine is simple, rapid and accurate, which is suitable for the identification of aconitine in the cases with suspected aconitine posioning in medical and forensic science.

Aconitine

Linear range

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Determination of Metabolites of Acrolein and Crotonaldehyde in Human Urine by Liquid Chromatography with Tandem Mass Spectrometry

Tobacco Science & Technology (2009)

Xue Yang Zhao Ge Fuwei Xie Wang Sheng Xie Jian-Pin

A method based on liquid chromatography with tandem mass spectrometry(HPLC-MS/MS) was developed for the simultaneous determination of 3-hydroxypropyl mercapturic acid(3-HPMA) and 3-hydroxy-1-methyl-propylmercapturic acid(3-HMPMA),the mercapturic acid metabolites of acrolein and crotonaldehyde in human urine.Using 3-HPMA-D3 and 3-HMPMA-D3 as internal standards,the urine samples were cleaned up by Isolute ENV+ solid phase extraction(SPE) cartridge and separated by an Angilent Extend C18 column,and then 3-HPMA and 3-HMPMA in the samples were detected by the tandem mass detector under multi-reaction monitoring scan mode.The contents of 3-HPMA and 3-HMPMA in urine samples from eight smokers and eight nonsmokers were determined,the results showed that:1) the recoveries of 3-HPMA and 3-HMPMA ranged from 91.11% to 114.66% with the relative standard deviations(RSDs) of 2.41% to 11.22%,and the limit of detection was of 1.0 and 1.3 ng/mL,respectively;2) the contents of 3-HPMA and 3-HMPMA in urine samples from eight smokers ranged from 943.05 to 6303.54 and from 1061.73 to 6373.74 ng/mL,and from eight nonsmokers ranged from 14.13 to 640.93 and from 23.36 to 119.40 ng/mL,respectively.

Crotonaldehyde

Mercapturic acid

Solid phase extraction

Acrolein

10.3969/j.issn.1002-0861.2009.08.009

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Purification of the recombinant green fluorescent protein from tobacco plants using alcohol/salt aqueous two-phase system and hydrophobic interaction chromatography

Research Square (Research Square) (2019)

Jie Dong Xiangzhen Ding Wang Sheng

Abstract Background: The green fluorescent protein (GFP) has been regarded as a valuable tool and widely applied as a biomarker in medical applications and diagnostics. A cost-efficient upstream expression system and an inexpensive downstream purification process will meet the demands of the GFP protein with high-purity. Results : The recombinant GFP was transiently expressed in an active form in agoinoculated N. benthamiana leaves by using Tobacco mosaic virus (TMV) RNA-based overexpression vector (TRBO). The yield of recombinant GFP was up to ~60% of total soluble proteins (TSP). Purification of recombinant GFP from the clarified lysate of N. benthaniana leaves was achieved by using an alcohol/salt aqueous two-phase system (ATPS) and following with a further hydrophobic interaction chromatography (HIC). The purification process takes only ~4 h and can recover 34.1 % of the protein. The purity of purified GFP was more than 95% and there were no changes in its spectroscopic characteristics. Conclusions: The strategy described here combines the advantages of both the economy and efficiency of plant virus-based expression platform and the simplicity and rapidity of environmentally friendly alcohol/salt ATPS. It has a considerable potential for the development of a cost-efficient alternative for production of recombinant GFP. Keywords: Green fluorescent protein, Plant virus, Transient gene expression, Aqueous two-phase system, Hydrophobic interaction chromatography

Downstream processing

10.21203/rs.2.12829/v3

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[Effects of seven RNA silencing suppressors on heterologous expression of green fluorescence protein expression mediated by a plant virus-based system in Nicotiana benthamiana].

PubMed (2012)

Wang Sheng Jie Dong Min Cao MU Hong-zhen Guoping Ding

To test the effects of 7 virus-encoded RNA silencing suppressors (RSSs) for enhancement of a plant virus-based vector system-mediated heterologous expression of green fluorescence protein (GFP) in Nicotiana benthamiana.Seven transient expression vectors for the 7 RSSs were constructed and co-inoculated on the leaves of Nicotiana benthamiana with PVXdt-GFP vector, a novel Potato virus X-based plant expression vector, through agroinfiltration. The protein and mRNA expression levels of the reporter gene GFP in the co-inoculated Nicotiana leaves were examined by Western blotting, ELISA and RT-qPCR to assess the effect of the RSSs for GFP expression enhancement.The 7 RSSs differed in the degree and duration of enhancement of heterologous GFP expression, and the p19 protein of Tomato bushy stunt virus (TBSV) induced the highest expression of GFP. African cassava mosaic virus AC2 protein and Rice yellow mettle virus P1 protein produced no obvious enhancement GFP expression.Transient co-expression of RSSs suppresses host silencing response to allow high-level and long-term expression of heterologous genes in plant, but the optimal RSS has to be identified for each plant virus-based expression vector system.

Agroinfiltration

Potato virus X

Heterologous expression

Heterologous

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Analysis of the Heterocyclic Aromatic Amines in Cigarette Smoke by Liquid Chromatography–Tandem Mass Spectrometry

Chromatographia (2014)

Ge Zhao Wang Sheng Yufeng Fu Jingjing Yu Bing Wang

Matrix (chemical analysis)

Solid phase extraction

10.1007/s10337-014-2687-8

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Analysis of Several Aromatic Amines in Cigarette Smoke by GC/MS with Selection Ion Monitoring Mode

Tobacco Science & Technology (2008)

Xia Qiao-Ling Fuwei Xie Wang Sheng

A GC/MS method for determining 1-aminonaphthalene,2-aminonaphthalene,3-amino-biphenyl,and 4-amino-biphenyl in cigarette smoke was developed.The cigarette smoke was collected by Cambridge filter pad,then the pad were extracted with 5%HCl,the extract was washed with methyl dichloride,alkalized by sodium hydroxide solution,re-extracted with n-hexane,derived with pentafluoride propionyl anhydride,purified by a Florisil column,and determined by GC/MS with selection ion monitoring mode using d3-2-aminonaphthalene as an internal standard.The results showed that the recoveries of the 4 aromatic amines were over 90% with RSD of below 10% and the limits of detection of below 0.7 ng/mL.The method is suitable for the quantitative analysis of the 4 aromatic amines in cigarette smoke.

Sodium hydroxide

Biphenyl

Selected ion monitoring

10.3969/j.issn.1002-0861.2008.11.011

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Higher-Order RNA Structural Requirements and Small-Molecule Induction of Tombusvirus Subgenomic mRNA Transcription

Journal of Virology (2008)

Wang Sheng Leyla Mortazavi K. Andrew White

Subgenomic (sg) mRNAs are small viral messages that are synthesized by polycistronic positive-strand RNA viruses to allow for the translation of certain viral proteins. Tombusviruses synthesize two such sg mRNAs via a premature termination mechanism. This transcriptional process involves the viral RNA-dependent RNA polymerase terminating minus-strand RNA synthesis prematurely at internal RNA signals during copying of the viral genome. The 3'-truncated minus-strand RNAs generated by the termination events then serve as templates for sg mRNA transcription. A higher-order RNA structure in the viral genome, located just upstream from the termination site, is a critical component of the RNA-based polymerase attenuation signal. Here, we have analyzed the role of this RNA structure in mediating efficient sg mRNA2 transcription. Our results include the following: (i) we define the minimum overall thermodynamic stability required for an operational higher-order RNA attenuation structure; (ii) we show that the distribution of stability within an attenuation structure affects its function; (iii) we establish that an RNA quadruplex structure can act as an effective attenuation structure; (iv) we prove that the higher-order RNA structure forms and functions in the plus strand; (v) we provide evidence that a specific attenuation structure-binding protein factor is not required for transcription; (vi) we demonstrate that sg mRNA transcription can be controlled artificially through small-molecule activation using RNA aptamer technology. These findings provide important new insights into the premature termination mechanism and present a novel approach to regulate the transcriptional process.

Subgenomic mRNA

Transcription

RNA-induced transcriptional silencing

Nucleic acid structure

Small nuclear RNA

RNA Silencing

10.1128/jvi.02416-07

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Determination of Organic Chlorine Pesticide in Soil Samples by GC

Wang Sheng

BHC,aldrin,dieldrin,endosulfanⅠ,endosulfanⅡ,endosulfan sulfate,endrin,endrinaldehyde,heptachlor,heptachlor epoxide isomer B and DDT in soil were determined by GC-ECD with Accelerated Solvent Extraction.The method is a rapid,accurate and effective for the determination of Organic chlorine pesticide in soil.

Endrin

Aldrin

Heptachlor

Endosulfan

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Determination of Eight Volatile Carbonyl Compounds in Cigarette Filter by LC-ESI-MS/MS

Tobacco Science & Technology (2013)

Jingjing Yu Wang Sheng Bing Wang Xiaodong Zhao Cai Junlan

Volatile organic compound

10.3969/j.issn.1002-0861.2013.09.010

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Advances in Molecular Biology of Tomato bushy stunt virus

Life Science Research (2013)

Wang Sheng

Plant virus RNA-based vectors have been developed for expressing recombinant proteins,and studying plant virology and phytopathology in recent years.Tomato bushy stunt virus(TBSV) is the type species of the plant-infecting Tombusvirus genus in the family Tombusviridae.Important advances were made in the molecular biology of TBSV,such as gene expression,genome replication,and recombination.Those advances in TBSV make it possible to be a candidate of plant virus RNA-based expression vector.

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