1.Alginate lyase immobilized Chlamydomonas algae microrobots: minimally invasive therapy for biofilm penetration and eradication.
Xiaoting ZHANG ; Huaan LI ; Lu LIU ; Yanzhen SONG ; Lishan ZHANG ; Jiajun MIAO ; Jiamiao JIANG ; Hao TIAN ; Chang LIU ; Fei PENG ; Yingfeng TU
Acta Pharmaceutica Sinica B 2025;15(6):3259-3272
Bacterial biofilms can make traditional antibiotics impenetrable and even promote the development of antibiotic-resistant strains. Therefore, non-antibiotic strategies to effectively penetrate and eradicate the formed biofilms are urgently needed. Here, we demonstrate the development of self-propelled biohybrid microrobots that can enhance the degradation and penetration effects for Pseudomonas aeruginosa biofilms in minimally invasive strategy. The biohybrid microrobots (CR@Alg) are constructed by surface modification of Chlamydomonas reinhardtii (CR) microalgae with alginate lyase (Alg) via biological orthogonal reaction. By degrading the biofilm components, the number of CR@Alg microrobots with fast-moving capability penetrating the biofilm increases by around 2.4-fold compared to that of microalgae. Massive reactive oxygen species are subsequently generated under laser irradiation due to the presence of chlorophyll, inherent photosensitizers of microalgae, thus triggering photodynamic therapy (PDT) to combat bacteria. Our algae-based microrobots with superior biocompatibility eliminate biofilm-infections efficiently and tend to suppress the inflammatory response in vivo, showing huge promise for the active treatment of biofilm-associated infections.
2.Cloning,fusion expression and identification of thioredoxin encoding gene from Toxoplasma gondii
Zigang ZHANG ; Xiaomei CHEN ; Danhua SU ; Yuan LIU ; Tao FU ; Jiamiao DUANMU ; Liang WU ; Xugan JIANG ; Shengxia CHEN ; Jianping CAO
Chinese Journal of Schistosomiasis Control 2016;28(3):289-292
Objective To clone and express the thioredoxin(Trx)from RH strain tachyzoites of Toxoplasma gondii,estab?lish the prokaryotic expression vector and purify the recombinant protein,then produce the polyclonal anti?Trx antibody in rab?bits. Methods Trx fragment was amplified by PCR and cloned into the pET?28a(+)vector,and the recombinant protein was in?duced with IPTG and purified by Ni?NTA affinity chromatography. The polyclonal antibody specificity was detected by Western blotting. Results The trx gene was amplified from T. gondii cDNA by PCR. The recombinant plasmid trx/pET?28a(+)was use?fully constructed,and the recombinant TRX protein was expressed and purified. The TRX polyclonal antibody was also ob?tained. The specific band of TRX was detected by Western blotting. Conclusion Western blotting can detect the specificity of polyclonal anti?Trx antibody,which will facilitate the biological functions of Trx.
3.Optimization of processing parameters for extraction of chlorogenic acid in Fule Granula using Box-Behnken experimental design
Chinese Journal of Biochemical Pharmaceutics 2016;36(4):196-198,202
Objective To optimize the processing parameters for the extraction of chlorogenic acid in Fule Granula by Box-Behnken experimental design.Methods The content of chlorogenic acid in Fule Granula was determination.The separation was performed on a Agilent ZORBAX Eclipse XDB C18 column(4.6 mm ×250 mm, 5 μm), and column temperature 30 ℃.Aetonitrile-0.2% H3PO4 was used as the mobile phase.The detection wavelength was at 327 nm.A three-factor and three-level Box-Behnken experimental design was employed to investigate effects of extraction time, solid-liquid ratio and extraction times on composite score of extracting amount of chlorogenic acid.Results Optimum process conditions were as follows:12 times the amount of water extracted three times, each time 60 min; chlorogenic acid extraction rate of 3.379%.Conclusion This optimized extraction technology has good predictability by Box-Behnken experimental design, and overall desirability, it provides a reference for application of Fule Granula.

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