1.Establishment of a duplex real-time PCR method for differentiation of African swine fever virus I177L gene-deleted strains
Yanxing LIN ; Peng XU ; Weijun SHI ; Chaohua HUANG ; Qiaoyu WENG ; Jiang WU ; Zhouxi RUAN ; Caihong ZHANG ; Chenfu CAO ; Junxing YANG ; Ye JIN ; Peng CHEN ; Qunyi HUA
Chinese Journal of Veterinary Science 2024;44(9):1848-1853
African swine fever virus(ASFV)I177L gene deletion vaccine is one of the key directions of African swine fever(ASF)live attenuated vaccine research and development.In order to effec-tively distinguish between the wild-type ASFV strain and the I177L gene-deleted strain,specific primers and probes were designed based on ASFV B646L and I177L genes,respectively.After screening and optimization,a duplex real-time PCR method was developed that can simultaneously detect these two genes.The results showed that ASFV B646L and I177L genes were detected spe-cifically and simultaneously by the method developed without cross-reactions with porcine circovir-us type 2,Seneca virus A,classical swine fever virus,foot-and-mouth disease virus,porcine respira-tory and reproductive syndrome virus.The detection limits of the duplex real-time PCR for recom-binant plasmids pUC57-B646L and pUC57-I177L were 1×103 copies/mL.The intra-and inter-as-say coefficients of variation were less than 4%,respectively.Detection of 122 pork and pork prod-ucts using the duplex real-time PCR developed and the real-time PCR recommended by WOAH showed that the coincidence rates of the two methods for B646L gene detection was 100%with two amplification curves appeared in the positive results of the established methods.The method established in this study can be used for the detection of ASFV I177L gene deletion strains,which provides technical support for ASF surveillance and epidemiological investigation.
2.Repurposing clinical drugs is a promising strategy to discover drugs against Zika virus infection.
Weibao SONG ; Hongjuan ZHANG ; Yu ZHANG ; Rui LI ; Yanxing HAN ; Yuan LIN ; Jiandong JIANG
Frontiers of Medicine 2021;15(3):404-415
Zika virus (ZIKV) is an emerging pathogen associated with neurological complications, such as Guillain-Barré syndrome in adults and microcephaly in fetuses and newborns. This mosquito-borne flavivirus causes important social and sanitary problems owing to its rapid dissemination. However, the development of antivirals against ZIKV is lagging. Although various strategies have been used to study anti-ZIKV agents, approved drugs or vaccines for the treatment (or prevention) of ZIKV infections are currently unavailable. Repurposing clinically approved drugs could be an effective approach to quickly respond to an emergency outbreak of ZIKV infections. The well-established safety profiles and optimal dosage of these clinically approved drugs could provide an economical, safe, and efficacious approach to address ZIKV infections. This review focuses on the recent research and development of agents against ZIKV infection by repurposing clinical drugs. Their characteristics, targets, and potential use in anti-ZIKV therapy are presented. This review provides an update and some successful strategies in the search for anti-ZIKV agents are given.
Adult
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Animals
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Drug Repositioning
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Humans
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Infant, Newborn
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Microcephaly
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Pharmaceutical Preparations
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Zika Virus
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Zika Virus Infection/prevention & control*
3.Identification of anti-Mycobacterium tuberculosis agents targeting the interaction of bacterial division proteins FtsZ and SepFe.
Hongjuan ZHANG ; Ying CHEN ; Yu ZHANG ; Luyao QIAO ; Xiangyin CHI ; Yanxing HAN ; Yuan LIN ; Shuyi SI ; Jiandong JIANG
Acta Pharmaceutica Sinica B 2023;13(5):2056-2070
Tuberculosis (TB) is one of the deadly diseases caused by Mycobacterium tuberculosis (Mtb), which presents a significant public health challenge. Treatment of TB relies on the combination of several anti-TB drugs to create shorter and safer regimens. Therefore, new anti-TB agents working by different mechanisms are urgently needed. FtsZ, a tubulin-like protein with GTPase activity, forms a dynamic Z-ring in cell division. Most of FtsZ inhibitors are designed to inhibit GTPase activity. In Mtb, the function of Z-ring is modulated by SepF, a FtsZ binding protein. The FtsZ/SepF interaction is essential for FtsZ bundling and localization at the site of division. Here, we established a yeast two-hybrid based screening system to identify inhibitors of FtsZ/SepF interaction in M. tuberculosis. Using this system, we found compound T0349 showing strong anti-Mtb activity but with low toxicity to other bacteria strains and mice. Moreover, we have demonstrated that T0349 binds specifically to SepF to block FtsZ/SepF interaction by GST pull-down, fluorescence polarization (FP), surface plasmon resonance (SPR) and CRISPRi knockdown assays. Furthermore, T0349 can inhibit bacterial cell division by inducing filamentation and abnormal septum. Our data demonstrated that FtsZ/SepF interaction is a promising anti-TB drug target for identifying agents with novel mechanisms.
4.Erratum to "Identification of anti-Mycobacterium tuberculosis agents targeting the interaction of bacterial division proteins FtsZ and SepFe" Acta Pharmaceutica Sinica B 13 (2023) 2056-2070.
Hongjuan ZHANG ; Ying CHEN ; Yu ZHANG ; Luyao QIAO ; Xiangyin CHI ; Yanxing HAN ; Yuan LIN ; Shuyi SI ; Jiandong JIANG
Acta Pharmaceutica Sinica B 2023;13(7):3180-3180
[This corrects the article DOI: 10.1016/j.apsb.2023.01.022.].