1.Prokaryotic expression,purification,and in vivo and in vitro interaction studies of the potential virulence factors EsxA and EsxB of Mycobacterium haemophilum
Rongxian XIE ; Longyun CHENG ; Xilu YUAN ; Li LI ; Haihong JIA ; Bingqing LI
Chinese Journal of Zoonoses 2025;41(8):824-831
This studyobtained the proteins of virulence factors EsxA and EsxB from Mycobacterium haemophilum and explored their interactions both in vitro and in vivo.The aim was to lay a foundation for further analysis of the functional mechanisms of EsxA and EsxB,and to further the development of drugs targeting Mycobacterium haemophilum.On the basis of bioinformatics analysis of the virulence factors EsxA and EsxB from Mycobacterium haemophilum,we performed molecular cloning,using Mycobacterium haemophi-lum as a template to construct recombinant plasmids EsxA-pGl01,EsxB-pGl01,and EsxB-pET-29b.The proteins of EsxA,EsxB,and their complex were expressed with a prokaryotic system,then purified through nickel-affinity chromatography and gel filtration chromatography.Intracellular colocalization was determined through fluorescence colocalization.Prokaryotic expression systems for EsxA,EsxB,and their complex were successfully constructed,and purified proteins were obtained.Fluorescence colocalization indi-cated that EsxA and EsxB interacted in vivo.In vivo fluorescence colocalization and in vitro complex formation suggested that EsxA and EsxB interacted both intracellularly and extracellularly.Our findings lay a foundation for studying the pathogenic mechanisms of the virulence factors EsxA and EsxB in Mycobacterium haemophilum,and performing structural analysis of their complex.
2.Prokaryotic expression,purification,and in vivo and in vitro interaction studies of the potential virulence factors EsxA and EsxB of Mycobacterium haemophilum
Rongxian XIE ; Longyun CHENG ; Xilu YUAN ; Li LI ; Haihong JIA ; Bingqing LI
Chinese Journal of Zoonoses 2025;41(8):824-831
This studyobtained the proteins of virulence factors EsxA and EsxB from Mycobacterium haemophilum and explored their interactions both in vitro and in vivo.The aim was to lay a foundation for further analysis of the functional mechanisms of EsxA and EsxB,and to further the development of drugs targeting Mycobacterium haemophilum.On the basis of bioinformatics analysis of the virulence factors EsxA and EsxB from Mycobacterium haemophilum,we performed molecular cloning,using Mycobacterium haemophi-lum as a template to construct recombinant plasmids EsxA-pGl01,EsxB-pGl01,and EsxB-pET-29b.The proteins of EsxA,EsxB,and their complex were expressed with a prokaryotic system,then purified through nickel-affinity chromatography and gel filtration chromatography.Intracellular colocalization was determined through fluorescence colocalization.Prokaryotic expression systems for EsxA,EsxB,and their complex were successfully constructed,and purified proteins were obtained.Fluorescence colocalization indi-cated that EsxA and EsxB interacted in vivo.In vivo fluorescence colocalization and in vitro complex formation suggested that EsxA and EsxB interacted both intracellularly and extracellularly.Our findings lay a foundation for studying the pathogenic mechanisms of the virulence factors EsxA and EsxB in Mycobacterium haemophilum,and performing structural analysis of their complex.
3.Development and application of emergency management information systems for medical radiation
Ying CHENG ; Longyun WAN ; Xing WU
Chinese Journal of Radiological Health 2022;31(5):635-638
High-energy radiation derived from X-ray, γ-ray, neutrons, and other radioisotopes has been widely used for disease diagnosis and treatmentin clinical practice. Notably, high-energy radiation has been proven to increase the cure rate, prolong the survival time, and improve the quality of life among patients with malignant tumors. However, radiation poses huge threats to human health and life. Establishment of effective emergency management information systems for medical radiation is therefore of great significance to evaluate the radiation safety, predict the leakage of radioactive materials, and propose effective responses. This review summarizes the development and application of currently main emergency management information systems for medical radiation, so as to provide a reference for the establishment of sensitive and effective hospital-based radiation emergency management information systems.
4.Quality classification standard of Aucklandia lappa seed.
Yu WANG ; Xue ZHANG ; Daxi CHEN ; Longyun LI ; Taiqing CHENG
China Journal of Chinese Materia Medica 2012;37(14):2052-2054
OBJECTIVETo establish a quality classification standard for Aucklandia lappa.
METHODSeed purity, 1 000-grain weight, germination rate, water content of 37 seed samples from different areas were measured, and seed external characters were observed. The cluster analysis was applied for establishment of the primary quality classification standard of A. lappa seed.
RESULTThe seed quality grading of A. lappa was described as three grades. The seed quality of each grade should reach following requirements: for first grade seeds, seed germination rate > or = 85%, purity > or = 99%, 1 000-grain weight > or = 26.0 g, water content < or = 8.8%; for second grade seeds, seed germination rate > or = 70%, purity > or = 98%, 1 000-grain weight > or = 23.8 g, water content < or = 10%; for third grade seeds, seed germination rate > or = 50%, purity > or = 96%, 1 000-grain weight > or = 23.0 g, water content < or = 11%.
CONCLUSIONThis quality classification standard of A. lappa can be used as the quality control standard of A. lappa.
Asteraceae ; growth & development ; Plants, Medicinal ; growth & development ; Quality Control ; Seeds ; physiology
5.DNA Extraction from Rhizoma Coptidus and optimization of RAPD reaction system
Daxia CHEN ; Longyun LI ; Min QIAN ; Cheng LU
Chinese Traditional and Herbal Drugs 1994;0(08):-
Objective To study the genomic DNA extraction from Rhizoma Coptidus and optimization of RAPD reaction system. Methods Different methods, i.e. phenol method, CTAB method, low pH extraction medium with high salt, were used to genomic DNA extract from Rhizoma Coptidus. The DNA samples obtained by the above methods were tested by agarose gel electrophoresis and ultraviolet spectrometer. Results CTAB Method was considered to be an optimal technique. Based on the genomic DNA extracted by CTAB method, a reaction system suitable for Rhizoma Coptidus was established, that is, 25 ?L amplification reactions system containing 1?PCR buffer, 2 mmol/L Mg 2+, 100—150 ?mol/L dNTP, 20 ng primer, 40 ng template DNA, and 1 U Taq DNA polymerase. Conclusion CTAB Method and RAPD reaction system can be used to RAPD analysis in Rhizoma Coptidus.

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