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Characterization of a Novel Gene, srpA, Conferring Resistance to Streptogramin A, Pleuromutilins, and Lincosamides in Streptococcus suis

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Engineering 2022年第2期8卷 85-94页

作者： Chaoyang Zhang Lu Liu Peng Zhang Jingpo Cui Xiaoxia Qin Lichao Ma Kun Han Zhanhui Wang Shaolin Wang Shuangyang Ding Zhangqi Shen College of veterinary Medicine China Agricultural UniversityBeijing 100193China Beijing key laboratory of Detection Technology for Animal-Derived Food safety and Beijing laboratory for Food Quality and safety China Agricultural UniversityBeijing 100193China

Antimicrobial resistance is undoubtedly one of the greatest global health threats. The emergence of multidrug-resistant(MDR) Gram-positive pathogens, such as methicillin-resistant Staphylococcus aureus(MRSA), vancomycin-resistant Enterococcus faecium(VRE), and β-lactamase-resistant Streptococcus pneumonia, has severely limited our antibiotic arsenal. Numerous ribosome-targeting antibiotics, especially pleuromutilins, oxazolidinones, and streptogramins, are viewed as promising alternatives against aggressive MDR pathogens. In this study, we identified a new adenosine triphosphate(ATP)-binding cassete(ABC)-F family determinant, srp A, in Streptococcus suis(S. suis) by means of a comparative analysis of the whole-genome sequences of tiamulin(TIA)-resistant and TIA-sensitive bacteria. Functional cloning confirmed that the deduced gene can mediate cross-resistance to pleuromutilins, lincosamides, and streptogramin A in S. suis and S. aureus. A sequence alignment revealed that Srp A shares the highest amino acid identity with Vga(E)(36%) and shows canonical characteristics of ABC-F family *** Srp A-ribosome docked compounds, the extended loop region of Srp A approaches the valnemulinbinding pocket in the ribosome peptidyl-transferase center and competes with bound valnemulin. A detailed mutational analysis of the loop residues confirmed that this domain is crucial for Srp A activity,as substitutions or truncations of this region affect the efficiency and specificity of antibiotic *** antibiotics accumulation indicated that Srp A does not act as an efflux pump, while a ribosome binding assay supported the protective effects of Srp A on the ribosome by preventing antibiotic binding as well as displacing bound drugs. These findings clarify the mechanisms underlying resistance to ribosomal antibiotics.

关键词： SrpA Streptococcus suis Antibiotic resistance Ribosome ABC-F family proteins

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Syntheses,Crystal Structures and Antibacterial evaluation of Two New Pleuromutilin Derivatives

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Chinese Journal of Structural Chemistry 2016年第4期35卷 529-536页

作者：尚若锋许水金衣云鹏艾鑫梁剑平 key laboratory of New Animal drug Project of Gansu Province key laboratory of veterinary Pharmaceutical Development Ministry of Agriculture Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS Yancheng Youhua Pharmaceutical & Chemical Technology Co. Ltd.

Two new pleuromutilin derivatives, 14-O-[（4-amino-6-methoxyl-pyrimidine-2-yl）-thioacetyl] mutilin（4） and 14-O-[4-amino-1-methyl-6-oxo-1,6-dihydropyrimidin-2-yl）thioacetyl] mutilin（5）, were synthesized and structurally characterized by IR, NMR spectra, HRMS and single-crystal X-ray diffraction. These compounds contain a 5-6-8 tricyclic carbon skeleton and a pyrimidine ring. Compound 4 is in the monoclinic system, space group P1211 with a = 10.2517（4）, b = 12.5655（4）, c = 10.3435（4）A, V = 1315.69（8） A^3, Z = 2, Dc = 1.309 g/cm3, F（000） = 558, μ = 0.166 mm^-1, S = 1.047, R = 0.0457 and w R = 0.0934 for 4721 unique reflections（R（int） = 0.0322） with I 〉 2σ（I）. Compound 5 belongs to the orthorhombic system, space group P212121 with a = 7.3667（4）, b = 13.9990（7）, c = 29.0434（13） A, V = 2995.1（2） A^3, Z = 4, Dc = 1.250 g/cm^3, F（000） = 1216, μ = 0.153 mm^-1, S = 1.031, R = 0.0545 and wR = 0.0982 for 5242 unique reflections（R（int） = 0.0476） with I 〉 2σ（I））. The in vitro antibacterial activity study showed the title compounds 4 and 5 displayed slightly less activity against methicillin-resistant Staphylococcus aureus（MRSA） and methicillin-resistant Staphylococcus epidermidis（MRSE）, and lower potent against Escherichia coli（***） and Bacillus subtilis（***） when compared to those of tiamulin fumarate.

关键词： pleuromutilin single-crystal structure antibacterial activities

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A seven-gene-deleted African swine fever virus is safe and effective as a live attenuated vaccine in pigs

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Science China(Life Sciences) 2020年第5期63卷 623-634页

作者： Weiye Chen Dongming Zhao Xijun He Renqiang Liu Zilong Wang Xianfeng Zhang Fang Li Dan Shan Hefeng Chen Jiwen Zhang Lulu Wang Zhiyuan Wen Xijun Wang Yuntao Guan Jinxiong Liu Zhigao Bu State key laboratory of veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150001China National High Containment laboratory for Animal Diseases Control and Prevention Harbin 150069China

African swine fever(ASF)is a devastating infectious disease in swine that is severely threatening the global pig *** efficacious vaccine is urgently ***,we used the Chinese ASFV HLJ/18 as a backbone and generated a series of genedeleted *** virulence,immunogenicity,safety,and protective efficacy evaluation in specific-pathogen-free pigs,commercial pigs,and pregnant sows indicated that one virus,namely HLJ/18-7GD,which has seven genes deleted,is fully attenuated in pigs,cannot convert to the virulent strain,and provides complete protection of pigs against lethal ASFV *** study shows that HLJ/-18-7GD is a safe and effective vaccine against ASFV,and as such is expected to play an important role in controlling the spread of ASFV.

关键词： African swine fever vaccine safety protective efficacy pig

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Effects of 105 traditional Chinese medicines on the detection ofβ-agonists in medicine extracts and swine urine based on colloidal gold immunochromatographic assay

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Journal of Integrative Agriculture 2021年第6期20卷 1626-1635页

作者： WANG Zi-lin FENG Ke-ying GE Xiu-feng MAI Jia-cheng WANG Han-chuan LIU Wen-zi ZHANG Jia-hui SHEN Xiang-guang Guangdong Provincial key laboratory of veterinary Pharmaceutics Development and safety evaluation South China Agricultural UniversityGuangzhou 510642P.R.China Animal and Poultry Products Quality Control Inspection and Testing CenterMinistry of Agriculture and Rural AffairsGuangzhou 510642P.R.China

Colloidal gold immunochromatographic assay(CGIA)is commonly used for the on-site detection ofβ-agonists that are sometimes used illegally as feed additives in swine ***,few studies have evaluated the causes of false-positive results that sometimes occur when applying CGIA in agricultural *** this study,we investigated if this false-positive phenomenon is related to the addition of certain traditional Chinese medicines(TCMs)to swine *** established and verified an extraction method for TCMs,and then applied CGIA to detectβ-agonists in the extracts of 105 TCMs and in the urine of swine dosed with TCMs,*** chromatography-tandem mass spectrometry was used to validate the results of the urine samples tested positive forβ-agonists using *** results were also verified using TCMs and colloidal gold test strips produced by different *** extracts of Citri Reticulatae Pericarpium Viride,Citri Reticulatae Pericarpium,Magnoliae Officinalis Cortex,Chaenomelis Fructus,and Rhodiolae Crenulatae Radix Et Rhizoma were tested positive forβ-***,the addition of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium to swine feed resulted in false-positive results forβ-agonists in swine *** results provide a new way to explain false-positive CGIA results and provide valuable information for livestock feeding programs.

关键词： colloidal gold immunochromatographic assay false positive traditional Chinese medicine β-agonists swine urine

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evaluation of an attenuated vaccine candidate based on the genotype C of bovine parainfluenza virus type 3 in albino guinea pigs

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Journal of Integrative Agriculture 2017年第9期16卷 2047-2054页

作者： MA Lei ZHU Yuan-mao YANG Ting XUE Fei Division of Livestock Infectious Diseases State Key Laboratory of Veterinary BiotechnologyHarbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069P.R.China

Bovine parainfluenza virus type 3（BPIV3） is considered as one of the most important respiratory tract pathogens of both young and adult cattle, and widespread among cattle in the world. BPIV3 was first reported in China in 2008 and four strains of BPIV3 were isolated from Shandong Province, known as genotype C（BPIV3c）. Pathogen investigations had shown that BPIV3 c infection was very common among cattle in China. To date, BPIV3 can be classified into genotypes A, B and C based on genetic and phylogenetic analysis. Serological survey also demonstrates that BPIV3 infection is widespread in China, however, there is still no available vaccine for BPIV3 prevention in China nowadays. In the present study, the BPIV3 c strain SD0835 was continuously passaged on Madin-Darby bovine kidney（MDBK） cells for hundreds of times, and the pathogenicity of passage 209 was reduced in guinea pigs. The passage 209 of BPIV3 c strain SD0835 was used as a live vaccine candidate to immunize the guinea pigs. The vaccination results revealed that two vaccinations could induce excellent serum neutralizing antibody responses as well as proliferation of T lymphocytes. The vaccinated guinea pigs were well protected against challenge with a low passage of BPIV3 c strain SD0835. Additionally, the percentages of CD4~＋ and CD8~＋ T cell subsets of animals in vaccinated group increased after immunization; T cell subsets on day 2 after challenge in both groups decreased, and the decline of CD4~＋ and CD8~＋ T cell subsets levels of four guinea pigs in vaccinated group was relatively moderate, comparing with that of the control group. These data support further testing of the attenuated virus as an effective candidate vaccine.

关键词： bovine parainfluenza virus type 3 attenuated vaccine genotype C guinea pig

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G-quadruplexes in genomes of viruses infecting eukaryotes or prokaryotes are under different selection pressures from hosts

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Journal of Genetics and Genomics 2022年第1期49卷 20-29页

作者： Zhen Li Sheng Hu Qian Fan Wang Hany IMohamed Guangfu Yang Zhen-Xia Chen Dengguo Wei State key laboratory of Agricultural Microbiology College of Veterinary MedicineHuazhong Agricultural UniversityWuhanHubei 430070China Hubei Hongshan laboratory Huazhong Agricultural UniversityWuhan 430070China Interdisciplinary Sciences Institute Huazhong Agricultural UniversityWuhan 430070China College of Science Huazhong Agricultural UniversityWuhan 430070China College of Biomedicine and Health Huazhong Agricultural UniversityWuhan 430070China key laboratory of Pesticide and Chemical Biology Ministry of EducationCollege of ChemistryCentral China Normal UniversityWuhan 430079China International Joint research Center for Intelligent Biosensor Technology and Health Central China Normal UniversityWuhan 430079China Chemistry Department Faculty of ScienceBenha UniversityBenha 13518Egypt Hubei key laboratory of Agricultural Bioinformatics College of Life Science and TechnologyHuazhong Agricultural UniversityWuhan 430070China Shenzhen Institute of Nutrition and Health Huazhong Agricultural UniversityShenzhen 518124China Shenzhen Branch Guangdong Laboratory for Lingnan Modern AgricultureGenome Analysis Laboratory of the Ministry of AgricultureAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhen 518124China National Reference laboratory of veterinary drug residues(HZAU)and MAO key laboratory for Detection of veterinary drug residues Huazhong Agricultural UniversityWuhanHubei 430070China

G-quadruplexes in viral genomes can be applied as the targets of antiviral therapies, which has attracted wide interest. However, it is still not clear whether the pervasive number of such elements in the viral world is the result of natural selection for functionality. In this study, we identified putative quadruplex-forming sequences(PQSs) across the known viral genomes and analyzed the abundance, structural stability, and conservation of viral PQSs. A Viral Putative G-quadruplex Database(http://***/MBPC/Vi PGD/***/home/index) was constructed to collect the details of each viral PQS, which provides guidance for selecting the desirable PQS. The PQS with two putative G-tetrads(G2-PQS) was significantly enriched in both eukaryotic viruses and prokaryotic viruses, whereas the PQSs with three putative G-tetrads(G3-PQS) were only enriched in eukaryotic viruses and depleted in prokaryotic viruses. The structural stability of PQSs in prokaryotic viruses was significantly lower than that in eukaryotic viruses. Conservation analysis showed that the G2-PQS, instead of G3-PQS, was highly conserved within the genus. This suggested that the G2-quadruplex might play an important role in viral biology, and the difference in the occurrence of G-quadruplex between eukaryotic viruses and prokaryotic viruses may result from the different selection pressures from hosts.

关键词： G-quadruplex Selection pressure Database Evolution

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Acquisition of a novel conjugative multidrug-resistant hypervirulent plasmid leads to hypervirulence in clinical carbapenem-resistant Klebsiella pneumoniae strains

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mLife 2023年第3期2卷 317-327页

作者： Gong Li Ling Jia Lei Wan Lijuan Xia Ang Gao Runshi Yang Ruanyang Sun Minge Wang Juan Du Xinlei Lian Rongmin Zhang Liangxing Fang Xiaoping Liao Yahong Liu Bao-Tao Liu Jian Sun State key laboratory for Animal Disease Control and Prevention South China Agricultural UniversityGuangzhouChina Guangdong Provincial key laboratory of veterinary Pharmaceutics Development and safety evaluation South China Agricultural UniversityGuangzhouChina Department of Pathogen Biology and Microbiology Zhejiang University School of MedicineHangzhouChina College of veterinary Medicine Qingdao Agricultural UniversityQingdaoChina

The co-occurrence of plasmid-mediated multidrug resistance and hypervirulence in epidemic carbapenem-resistant Klebsiella pneumoniae has emerged as a global public health *** this study,an ST23 carbapenem-resistant hyper-virulent ***(CR-HvKP)strain VH1-2 was identified from cucumber in China and harbored a novel hybrid plasmid *** plasmid pVH1-2-VIR carrying both virulence and multidrug-resistance(MDR)genes was likely generated through the recombination of a virulence plasmid and an IncFIIK conjugative MDR plasmid in clinical ST2318622 isolated from a sputum *** plasmid pVH1-2-VIR exhibited the capacity for transfer to the clinical ST11 carbapenem-resistant ***(CRKP)strain via conjugation *** of pVH1-2-VIR plasmid directly converted a CRKP into CR-HvKP strain characterized by hypermucoviscosity,heightened virulence for Galleria mellonella larvae,and increased colonization ability in the mouse *** emergence of such a hybrid plasmid may expedite the spread of CR-HvKP strains,posing a significant risk to human health.

关键词： conjugative virulence plasmid CR-HvKP hybrid plasmid Klebsiella pneumoniae ST23

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The dose regimen formulation of doxycycline hydrochloride and florfenicol injection based on ex vivo pharmacokinetic-pharmacodynamic modeling against the Actinobacillus pleuropneumoniae in pigs

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Animal Diseases 2023年第4期3卷 286-298页

作者： Yuanyuan Yuan Boyu An Shuyu Xie Wei Qu Haihong Hao Lingli Huang Wanhe Luo Jixiang Liang Dapeng Peng National Reference laboratory of veterinary drug residues(HZAU)and MOA key laboratory for the Detection of veterinary drug residues in Foods Huazhong Agricultural UniversityWuhanHubei 430070People's Republic of China State key laboratory of Agricultural Microbiology Huazhong Agricultural UniversityWuhanHubei 430070People's Republic of China College of veterinary Medicine Huazhong Agricultural UniversityWuhan 430070HubeiPeople's Republic of China Shenzhen Institute of Nutrition and Health Huazhong Agricultural UniversityWuhan 430070HubeiPeople's Republic of China Shenzhen Branch Guangdong Laboratory for Lingnan Modern AgricultureGenome Analysis Laboratory of the Ministry of AgricultureAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhenPeople's Republic of China

Doxycycline hydrochloride and florfenicol combination(DoxHcl&FF)is an effective treatment for respiratory *** the study,our objective Was to evaluate the activity of DoxHcl&FF against Actinobacillus pleuropneumoniae(APP)in porcine pulmonary epithelial lining fluid(PELF)and the optimal dosage scheme to avoid the development of *** DoxHcl&FF Was administered intramuscularly(IM)at 20mg/kg,and the PELF was collected at differ-ent time *** minimum inhibitory concentration(MIC)and time-mortality curves were also included in the *** on the sigmoid Emax equation and dose equations,the study integrated the in vivo pharmacokinetic data of infected pigs and ex vivo pharmacodynamic data to obtain the area under concentration time curve(AUCo-24h)MIC values in PELF and achieve bacteriostatic activity,bactericidal activity and the virtual eradication of *** study showed that the combination of DoxHcl and FF caused no significant changes in PK *** peak concentration(Cmax)of FF in healthy and diseased pigs was 8.87±0.08 and 8.67±0.07μg/mL,the_AUCo-24h were.172.75±2.52 and 18022±3.13 h-μg/mL,the Cmax of DoxHcl was 7.91±0.09 and 7.99±0.05μg/mL,and the AUCo-24h was 129.96±3.70 h-μg/mL and 169.82±4.38 h-μg/***&FF showed strong concentra-tion-dependent *** bacteriostatic,bactericidal,and elimination activity were calculated as 5.61,18.83 and 32.68 h,and the doses were 1.37(bacteriostatic),4.59(bactericidal)and 7.99(elimination)mg/*** findings indicated that the calculated recommended dose could assist in achieving more precise administration,increasing the effectiveness of DoxHcl&FF treatment for APP infections.

关键词： Florfenicol Doxycycline hydrochloride PK-PD Pig Dose regimen Actinobacillus pleuropneumoniae

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Screening and evaluation of antioxidants from lees by micro-injector systems combined with a fluorescent probe,N-borylbenzyloxycarbonyl-3,7-dihydroxyphenoxazine,in living Drosophila

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Chinese Chemical Letters 2018年第10期29卷 1521-1527页

作者： Guangcui Chu Dandan Cheng Wenjuan Liu Xuefeng Wang Xiaotao Hou Yuanyuan Hou Hongjun Wei Jiagang Deng Gang Bai Tianjin University of Traditional Chinese Medidne Tianjin 300193 China State key laboratory of Medicinal Chemical Biology College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research Nankai University Tianjin 300353 China Guangxi Collaborative Innovation Center of Study on Functional Ingredients of Agricultural residues Guangxi Key Laboratory of Efficacy Study on Chinese Materia Medica Guangxi University of Chinese Medicine Nanning 530200 China JF-Pharmaceutical Technology Development Co. Ltd. Tianjin 300457 China

Over the past few decades, the determination of antioxidant activity by chemical probe has been widely reported, but in vivo evaluation via model organisms of Drosophila melanogaster and rapid discovery system has not been studied adequately. In this study, we determined the antioxidant activity of lees and demonstrated the ability of compounds in lees to scavenge H_2O_2 in vitro and in vivo by different chemical probes. Lees increased the ability of Drosophila against oxidative stress and antioxidant enzyme activity in vivo. Five ingredients of organic acids and flavones in lees extract that rapidly scavenged H_2O_2 were revealed by a post-column-derived HPLC-UV-FLD system based on 4-hydroxyphenylacetic acid（PHPAA）chemiluminescence. Additionally, another fluorescent probe,N-borylbenzyloxycarbonyl-3,7-dihydroxyphenoxazine（NBCD）, was selected to evaluate the reactive oxygen species（ROS） scavenging capacity of lees in living Drosophila melanogaster using a microfluidic injection test coupled with microscopic imaging analysis, and similar effects were observed in flies when they were treated with tartaric acid and caffeic acid. The results demonstrated that the novel integrated system was suitable for screening and evaluating antioxidant ingredients from natural products.

关键词： Lees Antioxidant Microinjection HPLC-UV-FLD NBCD PHPAA

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Glioma cell membrane camouflaged cinobufotalin delivery system for combinatorial orthotopic glioblastoma therapy

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Nano research 2023年第8期16卷 11164-11175页

作者： Zibin Song Liqian Zhao Weiyi Fang Siyun Guo Anqi Xu Zhengming Zhan Yonghua Cai ShuaiShuai Xue Peng Chai Qiuhua Jiang Peng Zhao Ye Song Department of Neurosurgery Southern Medical University Nanfang HospitalGuangzhou 510515China Institute of Brain Diseases Nanfang Hospital of Southern Medical UniversityGuangzhou 510515China Cancer Center Integrated Hospital of Traditional Chinese MedicineSouthern Medical UniversityGuangzhou 510515China NMPA key laboratory for research and evaluation of drug Metabolism Guangdong Provincial Key Laboratory of Cardiac Function and Microcirculation GuangdongProvincial Key Laboratory of New Drug ScreeningSchool of Pharmaceutical SciencesSouthern Medical UniversityGuangzhou 510515China Department of Neurosurgery Ganzhou People's HospitalGanzhou 341000China

Glioblastoma(GBM)belongs to the deadliest primary malignancies with high mortality rate and poor *** the past decades,less progress has been made to treat GBM,owing largely to the lack of effective chemotherapeutics and poor drug accumulation in the glioma *** order to address this issue,we present an efficient biomimetic nanocomposite(Cu_(2−x)Se-CB@MEM,CCM),consisting of Cu_(2−x)Se nanoparticle core modified by cinobufotalin(CB),a toad venom extract,which is camouflaged with glioma cell Ln229 *** is demonstrated that CB can decrease the protein activity of inosine monophosphate dehydrogenase 1(IMPDH1),a key target correlated with prognosis,through intermolecular hydrogen bonding with amino acid residues ARG-105 and *** glioma cell membrane-camouflage endows the CCM with blood-brain barrier penetration and homology tumor-targeted *** optimized cinobufotalin based chemotherapy combining with the near-infrared-II(NIR-II)irradiation shows outstanding inhibition effect to glioma cells,by blocking cell cycle and inducing *** vivo mice bearing orthotopic Ln229 GBM treated with CCM+NIR-II(CCM+L)have significantly suppressed tumor growth and extended survival,without side *** glioma cell membrane camouflaged nanocomposite of Cu_(2−x)Se and cinobufotalin with its significant anti-glioma property and well biosafety will provide novel alternatives for clinical treatment of GBM.

关键词： glioblastoma cinobufotalin biomimetics photothermal therapy blood-brain barrier

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