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| 1 | Determining antimicrobial resistance profiles and identifying novel mutations of Neisseria gonorrhoeae genomes obtained by multiplexed MinION sequencing显示文摘Gonorrhea is one of the most common sexually transmitted diseases worldwide. To cure infection and prevent transmission,timely and appropriate antimicrobial therapy is necessary. Unfortunately, Neisseria gonorrhoeae, the etiological agent of gonorrhea, has acquired nearly all known mechanisms of antimicrobial resistance(AMR), thereby compromising the efficacy of antimicrobial therapy. Treatment failure resulting from AMR has become a global public health concern. Whole-genome sequencing is an effective method to determine the AMR characteristics of N. gonorrhoeae. Compared with next-generation sequencing, the MinION sequencer(Oxford Nanopore Technologies(ONT)) has the advantages of long read length and portability. Based on a pilot study using MinION to sequence the genome of N. gonorrhoeae, we optimized the workflow of sequencing and data analysis in the current study. Here we sequenced nine isolates within one flow cell using a multiplexed sequencing strategy. After hybrid assembly with Illumina reads, nine integral circular chromosomes were obtained. By using the online tool Pathogenwatch and a BLAST-based workflow, we acquired complete AMR profiles related to seven classes of antibiotics. We also evaluated the performance of ONT-only assemblies. Most AMR determinants identified by ONT-only assemblies were the same as those identified by hybrid assemblies. Moreover, one of the nine assemblies indicated a potentially novel antimicrobial-related mutation located in mtrR which results in a frame-shift, premature stop codon, and truncated peptide.In addition, this is the first study using the MinION sequencer to obtain complete genome sequences of N. gonorrhoeae strains which are epidemic in China. This study shows that complete genome sequences and antimicrobial characteristics of N.gonorrhoeae can be obtained using the MinION sequencer in a simple and cost-effective manner, with hardly any knowledge of bioinformatics required. More importantly, this strategy provides us with a potential approach to discover new AMR determinants. | Chi Zhang Feng Wang Cansheng Zhu Leshan Xiu Yamei Li Li Li Bo Liu Yizhun Li Yaling Zeng Boyang Guo Junping Peng | 2020 | Science China(Life Sciences)2020，63，7： | 2 |
| 2 | Highly Sensitive Spectrofluorimetric Determination of Riboflavin Based on the Generation of Active Oxygen Coupled with Enzymatic Reaction显示文摘A spectrofluorimetric method for the determining riboflavin (VB 2 ) based on its enhancement on the fluorescence of hemoglobin\|catalyzed enzymatic reaction was proposed. The proposed method consisted of two reactions. One was the photochemical reaction of VB 2 , the other was a hemoglobin\|catalyzed enzymatic reaction. The optimal experimental conditions for the determinations were established. The linear range of the method was 5.0×10 -9 \|1.0×10 -7 mol/L of VB 2 . The detection limit was calculated to be 3.65×10 -9 mol/L. The relative standard deviation of this method was 2.3 % at 7.0×10 -8 mol/L for 11 determinations. | Li Xiao\|yan 1,2 , Wang Rui\|yong 1 , Cai Ru\|xiu 1 ,Liu zhi\|hong 1 1. College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072,Hubei, China 2. Department of Chemistry, Leshan Teacher s College, Leshan, 61 | 2002 | Wuhan University Journal of Natural Sciences2002，7，3： | 1 |
| 3 | Efficient assembly of a large fragment of monkeypox virus genome as a qPCR template using dual-selection based transformation-associated recombination显示文摘Transformation-associated recombination(TAR)has been widely used to assemble large DNA constructs.One of the significant obstacles hindering assembly efficiency is the presence of error-prone DNA repair pathways in yeast,which results in vector backbone recircularization or illegitimate recombination products.To increase TAR assembly efficiency,we prepared a dual-selective TAR vector,pGFCS,by adding a PADH1-URA3 cassette to a previously described yeast-bacteria shuttle vector,p GF,harboring a PHIS3–HIS3 cassette as a positive selection marker.This new cassette works as a negative selection marker to ensure that yeast harboring a recircularized vector cannot propagate in the presence of 5-fluoroorotic acid.To prevent pGFCS bearing ura3 from recombining with endogenous ura3-52 in the yeast genome,a highly transformable Saccharomyces cerevisiae strain,VL6-48B,was prepared by chromosomal substitution of ura3-52 with a transgene conferring resistance to blasticidin.A55-kb genomic fragment of monkeypox virus encompassing primary detection targets for quantitative PCR was assembled by TAR using pGFCS in VL6-48B.The pGFCS-mediated TAR assembly showed a zero rate of vector recircularization and an average correct assembly yield of 79%indicating that the dual-selection strategy provides an efficient approach to optimizing TAR assembly. | Lei Yang Lingqian Tian Leshan Li Qiuhong Liu Xiang Guo Yuan Zhou Rongjuan Pei Xinwen Chen Yun Wang | 2022 | Virologica Sinica2022，37，3： | 1 |
| 4 | Research on Human Resources Development of Rural Public Service Departments in New Countryside Construction显示文摘On the basis of defining related concepts concerning human resources in rural public service departments,this paper analyses the overall status quo of human resources in China's rural public service departments and points out the problems existing in human resources in China's rural public service departments during new countryside construction as follows:first,the constitution structure is not rational;second,the cultural quality of staff is universally low,and the business capacity remains to be promoted;third,the improvement of human resources development environment lags behind,and the supporting reform is short.In the context of new countryside construction,the opportunities faced by the human resources in China's rural public service departments are as follows:China has elevated strengthening new countryside construction and rural talents construction as important state development strategy;the ideas of service-oriented government and learning-oriented government are put forward;civil servant system is overhauled.Therefore,I advance the development strategy of human resources in China's rural public service departments as follows:implement elastic personnel system reform in public service departments(including civil servant positions in department),to form the public service personnel system of 'able one comes in,mediocre one moves over and shiftless one steps down';audaciously promote young and middle-aged grass-roots cadres with strong business ability,high political quality,acute judgement and decision-making ability;build learning-oriented grass-roots public service organization,to make the staff in rural pubic service departments study assiduously and progress;broaden horizon,and build the human resources development system geared to international standards;in developed regions,the human resources in grass-roots public service departments should introduce enterprise competition model. | LI Zhong-dong Leshan Vocational and Technical College,Leshan 614000,China | 2011 | Asian Agricultural Research2011，3，8： | 0 |
| 5 | Comparison of viral propagation and drug response among SARS-CoV-2 VOCs using replicons capable of recapitulating virion assembly and release显示文摘Several variants of concern(VOCs)have emerged since the WIV04 strain of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)was first isolated in January 2020.Due to mutations in the spike(S)protein,these VOCs have evolved to enhance viral infectivity and immune evasion.However,whether mutations of the other viral proteins lead to altered viral propagation and drug resistance remains obscure.The replicon is a noninfectious viral surrogate capable of recapitulating certain steps of the viral life cycle.Although several SARS-CoV-2 replicons have been developed,none of them were derived from emerging VOCs and could only recapitulate viral genome replication and subgenomic RNA(sgRNA)transcription.In this study,SARS-CoV-2 replicons derived from the WIV04 strain and two VOCs(the Beta and Delta variants)were prepared by removing the S gene from their genomes,while other structural genes remained untouched.These replicons not only recapitulate viral genome replication and sgRNA transcription but also support the assembly and release of viral-like particles,as manifested by electron microscopic assays.Thus,the S-deletion replicon could recapitulate virtually all the post-entry steps of the viral life cycle and provides a versatile tool for measuring viral intracellular propagation and screening novel antiviral drugs,including inhibitors of virion assembly and release.Through the quantification of replicon RNA released into the supernatant,we demonstrate that viral intracellular propagation and drug response to remdesivir have not yet substantially changed during the evolution of SARS-CoV-2 from the WIV04 strain to the Beta and Delta VOCs. | Lingqian Tian Qiuhong Liu Rongjuan Pei Yingshan Chen Chonghui Xu Jielin Tang Hao Sun Kunpeng Liu Qi Yang Lei Yang Leshan Li Yongli Zhang Yuan Zhou Chao Shan Xue Hu Xinwen Chen Yun Wang | 2022 | Virologica Sinica2022，37，5： | 0 |
| 6 | Distribution threshold values of CaCl_2 onto the 10X-zeolite and macro-pore silica gel显示文摘The calcium chloride used for adsorption separation of ammonia is promising for its large adsorptive capacity and lower desorption temperature,but difficult to develop because of the liable expansion,lump and chip in the adsorption/desorption process.Composite adsorbents made by monolayer dis-persion of calcium chloride onto carriers with high surface areas exhibit better adsorptive capacity and stability.Several models were developed to confirm the maximum monolayer dispersion capacity of calcium chloride onto the carriers(the distribution threshold value),and the closely packed monolayer dispersion model was considered the most suitable for this study.The distribution threshold values given by this model were 0.60 g CaCl2/(g 10X-zeolite) and 0.38 g CaCl2/(g SiO2).When the divalent salt was dispersed onto the carriers,however,anions were separated into two types,causing that the en-tropy of the system tended to increase and the system was not stable.To minimize the entropy,a new model was put forward as the modified closely packed monolayer dispersion model.Based on this model,the distribution threshold values are 0.52 g CaCl2/(g 10X-zeolite) and 0.33 g CaCl2/(g SiO2),re-spectively.The distribution threshold values were also gained experimentally by XRD quantitative phase analysis:0.61 g CaCl2/(g 10X-zeolite) and 0.31 g CaCl2/(g SiO2).Comparison between experi-mental values of distribution threshold with theoretical ones based on two different model showed that the closely packed monolayer dispersion model fits the monolayer dispersion of calcium chloride onto micro-pore carrier - 10X-zeolite,and the modified closely packed monolayer dispersion model is more suitable for the bigger aperture carrier - macro-pore silica gel. | TANG YouCheng1,2,LI Bin1,YANG JingChang1,CHU Wei1 & LUO ShiZhong1 1 School of Chemical-Engineering,Sichuan University,Chengdu 610065,China 2 Sichuan Xinguang Silicon Technology Corporation Limited,Leshan 614000,China | 2009 | Science China Chemistry2009，52，2： | 0 |
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