[Objective] This study aimed to establish a Real-Time quantitative PCR method for the determination of transposon copy number in C. sakazakii. [ Method ] With single-copy housekeeping gene atpD as the reference gene, ...[Objective] This study aimed to establish a Real-Time quantitative PCR method for the determination of transposon copy number in C. sakazakii. [ Method ] With single-copy housekeeping gene atpD as the reference gene, recombinant plasmid containing both single-copy housekeeping gene atpD and EZ-TN5 transposon was constructed; based on the established standard curves for real-time quantitative detection of atpD gene and EZ-TN5 transposon, copy number of atpD gene and EZ-TN5 transpason in three C. sakazakii mutants was detected and the ratio was calculated. [ Result] Correlation coefficients of the standard curves for real-time quantitative detection of atpD gene and EZ-TN5 transposon were 0. 999 and 0.998, respectively ; the ratios of copy number of atpD gene and EZ-TN5 transposon in three C. sakazakii mutants were 0.98, 1.17 and 0.91, respectively, which indicates that EZ-TN5 transpeson in C. sakakii mutants is a single-copy. [ Conclusion] Real-time quantitative PCR method established in this study had high availability and could replace the Southern blot method to detect the copy num- ber of EZ-TN5 transposon in different bacteria.展开更多
Objective:Identifying novel strategies to prevent particulate matter(PM)-induced lung injury is crucial for the reduction of the morbidity of chronic respiratory diseases.The combined intervention represented by herba...Objective:Identifying novel strategies to prevent particulate matter(PM)-induced lung injury is crucial for the reduction of the morbidity of chronic respiratory diseases.The combined intervention represented by herbal formulae for simultaneously targeting multiple pathological processes can provide a more beneficial effect than the single intervention.The aim of this paper is therefore to design a safe and effective medicinal and edible Chinese herbs(MECHs)formula against PM-induced lung injury.Methods:PM-induced oxidative stress,inflammatory response and apoptosis A549 cell model were used to screen anti-oxidant,anti-inflammatory and anti-apoptotic MECHs,respectively.A network pharmacology method was utilized to rationally design a novel herbal formula.Ultra performance liquid chromatography-mass spectrometer was utilized to assess the quality control of MECHs formula.The excretion of magnetic iron oxide nanospheres of the MECHs formula was estimated in zebrafish.The MECH formula against PM-induced lung injury was investigated with mice experiments.Results:Five selected herbs were rationally designed to form a new MECH formula,including Citri Exocarpium Rubrum(Juhong),Lablab Semen Album(Baibiandou),Atractylodis Macrocephalae Rhizoma(Baizhu),Mori Folium(Sangye)and Polygonati Odorati Rhizoma(Yuzhu).The formula effectively promoted the magnetic iron oxide nanospheres excretion in zebrafish.The mid/high dose formula significantly prevented PM-induced lung damage in mice by enhancing the activity of SOD and GSH-Px,reducing the MDA and ROS level and attenuating the upregulation of pro-inflammatory cytokine(IL-6,IL-8,IL-1βand TNFa),down regulating the protein expression of NF-κB,STAT3 and Caspase-3.Conclusion:Our findings suggest that the effective MECHs formula will become a novel strategy for preventing PM-induced lung injury and provide a paradigm for the development of functional foods using MECHs.展开更多
Background:Bacterial infection,tissue hypoxia and inflammatory response can hinder infected wound repair.This study aimed to develop a multifunctional specific therapeutic photo activated release nanosystem[HMPB@MB@Au...Background:Bacterial infection,tissue hypoxia and inflammatory response can hinder infected wound repair.This study aimed to develop a multifunctional specific therapeutic photo activated release nanosystem[HMPB@MB@AuNPs@PMB@HA(HMAPH)]by loading photosen sitizer methylene blue(MB)into hollow mesoporous Prussian blue nanostructures and modifying the surface with gold particles,polymyxin B(PMB)and hydrophilic hyaluronic acid.Methods:The HMAPH was characterized using transmission electron microscopy,UV-vis,Fourier transform infrared spectroscopy,X-ray diffraction and X-ray photon spectroscopy.The photother mal performance,iron ion release and free radical generation of the HMAPH were measured under different conditions to investigate its thermo-responsive cascade reaction.The antibacterial ability of HMAPH was investigated using live/dead fluorescence tests.The morphology and membrane integrity of Pseudomonas aeruginosa(P.aeruginosa)were investigated using transmission electron microscopy.The anti-biofilm activity of HMAPH was evaluated using crystal violet and SYBR Green I staining.Finally,we established a mouse model of a skin wound infected by P.aeruginosa to confirm the in vivo effectiveness of HMAPH.We used immunofluorescent staining,hematoxylin-eosin staining,Masson staining and enzyme-linked immunosorbent assay to examine whether HMAPH promoted wound healing and reduced inflammatory damage.Results:In this study,hyaluronic acid was decomposed under the action of hyaluronidase.Also,the exposed nanomaterials specifically bound to the outer membrane of P.aeruginosa through PMB to increase the membrane sensitivity to photodynamic treatment.Under dual-light irradiation,a large amount of iron ions released by HMAPH underwent a Fenton reaction with H_(2)O_(2)in bacteria to generate hydroxyl radicals(·OH),enabling direct killing of cells by hyperthermia.Additionally,the photodynamic activity of MB released by photo-induced activation led to the generation of reactive oxygen species,achieving synergistic and effective inhibition of P.aeruginosa.HMAPH also inhibited biofilm formation and downregulated the expression of virulence factors.In vivo experiments revealed that HMAPH accelerated the healing of P.aeruginosa-infected wounds by promoting angiogenesis and skin regeneration,inhibiting the inflammatory response and promoting M1 to M2 polarization.Conclusions:Our study proposed a strategy against bacteria and biofilms through a synergistic photothermal-photodynamic-Fenton reaction,opening up new prospects for combating biofilmassociated infections.展开更多
Considering the increasing abundance of antibiotic-resistant bacteria,novel antimicrobial approaches need to be investigated.Photothermal therapy(PTT),an innovative noninvasive therapeutic technique,has demonstrated s...Considering the increasing abundance of antibiotic-resistant bacteria,novel antimicrobial approaches need to be investigated.Photothermal therapy(PTT),an innovative noninvasive therapeutic technique,has demonstrated significant potential in addressing drug-resistant bacteria and bacterial biofilms.However,when used in isolation,PTT requires higher-temperature conditions to effectively eradicate bacteria,thereby potentially harming healthy tissues and inducing new inflammation.This study aims to present a comprehensive review of nanomaterials with intrinsic antimicrobial properties,antimicrobial materials relying on photothermal action,and nanomaterials using drug delivery antimicrobial action,along with their applications in antimicrobials.Additionally,the synergistic mechanisms of these antimicrobial approaches are elucidated.The review provides a reference for developing multifunctional photothermal nanoplatforms for treating bacterially infected wounds.展开更多
基金Supported by Key Project of Tianjin Municipal Education Commission(2010ZD01)
文摘[Objective] This study aimed to establish a Real-Time quantitative PCR method for the determination of transposon copy number in C. sakazakii. [ Method ] With single-copy housekeeping gene atpD as the reference gene, recombinant plasmid containing both single-copy housekeeping gene atpD and EZ-TN5 transposon was constructed; based on the established standard curves for real-time quantitative detection of atpD gene and EZ-TN5 transposon, copy number of atpD gene and EZ-TN5 transpason in three C. sakazakii mutants was detected and the ratio was calculated. [ Result] Correlation coefficients of the standard curves for real-time quantitative detection of atpD gene and EZ-TN5 transposon were 0. 999 and 0.998, respectively ; the ratios of copy number of atpD gene and EZ-TN5 transposon in three C. sakazakii mutants were 0.98, 1.17 and 0.91, respectively, which indicates that EZ-TN5 transpeson in C. sakakii mutants is a single-copy. [ Conclusion] Real-time quantitative PCR method established in this study had high availability and could replace the Southern blot method to detect the copy num- ber of EZ-TN5 transposon in different bacteria.
基金funded by China Postdoctoral Science Foundation(2020T130093ZX)。
文摘Objective:Identifying novel strategies to prevent particulate matter(PM)-induced lung injury is crucial for the reduction of the morbidity of chronic respiratory diseases.The combined intervention represented by herbal formulae for simultaneously targeting multiple pathological processes can provide a more beneficial effect than the single intervention.The aim of this paper is therefore to design a safe and effective medicinal and edible Chinese herbs(MECHs)formula against PM-induced lung injury.Methods:PM-induced oxidative stress,inflammatory response and apoptosis A549 cell model were used to screen anti-oxidant,anti-inflammatory and anti-apoptotic MECHs,respectively.A network pharmacology method was utilized to rationally design a novel herbal formula.Ultra performance liquid chromatography-mass spectrometer was utilized to assess the quality control of MECHs formula.The excretion of magnetic iron oxide nanospheres of the MECHs formula was estimated in zebrafish.The MECH formula against PM-induced lung injury was investigated with mice experiments.Results:Five selected herbs were rationally designed to form a new MECH formula,including Citri Exocarpium Rubrum(Juhong),Lablab Semen Album(Baibiandou),Atractylodis Macrocephalae Rhizoma(Baizhu),Mori Folium(Sangye)and Polygonati Odorati Rhizoma(Yuzhu).The formula effectively promoted the magnetic iron oxide nanospheres excretion in zebrafish.The mid/high dose formula significantly prevented PM-induced lung damage in mice by enhancing the activity of SOD and GSH-Px,reducing the MDA and ROS level and attenuating the upregulation of pro-inflammatory cytokine(IL-6,IL-8,IL-1βand TNFa),down regulating the protein expression of NF-κB,STAT3 and Caspase-3.Conclusion:Our findings suggest that the effective MECHs formula will become a novel strategy for preventing PM-induced lung injury and provide a paradigm for the development of functional foods using MECHs.
基金supported by the National Key R&D Program of China(2023YFF1103900)National Natural Science Foundation of China(31902304,31972167)the Open Project Program of State Key Laboratory of Food Nutrition and Safety,Tianjin University of Science&Technology(SKLFNS-KF-202209)。
文摘Background:Bacterial infection,tissue hypoxia and inflammatory response can hinder infected wound repair.This study aimed to develop a multifunctional specific therapeutic photo activated release nanosystem[HMPB@MB@AuNPs@PMB@HA(HMAPH)]by loading photosen sitizer methylene blue(MB)into hollow mesoporous Prussian blue nanostructures and modifying the surface with gold particles,polymyxin B(PMB)and hydrophilic hyaluronic acid.Methods:The HMAPH was characterized using transmission electron microscopy,UV-vis,Fourier transform infrared spectroscopy,X-ray diffraction and X-ray photon spectroscopy.The photother mal performance,iron ion release and free radical generation of the HMAPH were measured under different conditions to investigate its thermo-responsive cascade reaction.The antibacterial ability of HMAPH was investigated using live/dead fluorescence tests.The morphology and membrane integrity of Pseudomonas aeruginosa(P.aeruginosa)were investigated using transmission electron microscopy.The anti-biofilm activity of HMAPH was evaluated using crystal violet and SYBR Green I staining.Finally,we established a mouse model of a skin wound infected by P.aeruginosa to confirm the in vivo effectiveness of HMAPH.We used immunofluorescent staining,hematoxylin-eosin staining,Masson staining and enzyme-linked immunosorbent assay to examine whether HMAPH promoted wound healing and reduced inflammatory damage.Results:In this study,hyaluronic acid was decomposed under the action of hyaluronidase.Also,the exposed nanomaterials specifically bound to the outer membrane of P.aeruginosa through PMB to increase the membrane sensitivity to photodynamic treatment.Under dual-light irradiation,a large amount of iron ions released by HMAPH underwent a Fenton reaction with H_(2)O_(2)in bacteria to generate hydroxyl radicals(·OH),enabling direct killing of cells by hyperthermia.Additionally,the photodynamic activity of MB released by photo-induced activation led to the generation of reactive oxygen species,achieving synergistic and effective inhibition of P.aeruginosa.HMAPH also inhibited biofilm formation and downregulated the expression of virulence factors.In vivo experiments revealed that HMAPH accelerated the healing of P.aeruginosa-infected wounds by promoting angiogenesis and skin regeneration,inhibiting the inflammatory response and promoting M1 to M2 polarization.Conclusions:Our study proposed a strategy against bacteria and biofilms through a synergistic photothermal-photodynamic-Fenton reaction,opening up new prospects for combating biofilmassociated infections.
基金supported by the National Key R&D Program of China(2023YFF1103900)National Natural Science Foundation of China(31902304,31972167)the Open Project Program of State Key Laboratory of Food Nutrition and Safety,Tianjin University of Science&Technology(SKLFNS-KF-202310).
文摘Considering the increasing abundance of antibiotic-resistant bacteria,novel antimicrobial approaches need to be investigated.Photothermal therapy(PTT),an innovative noninvasive therapeutic technique,has demonstrated significant potential in addressing drug-resistant bacteria and bacterial biofilms.However,when used in isolation,PTT requires higher-temperature conditions to effectively eradicate bacteria,thereby potentially harming healthy tissues and inducing new inflammation.This study aims to present a comprehensive review of nanomaterials with intrinsic antimicrobial properties,antimicrobial materials relying on photothermal action,and nanomaterials using drug delivery antimicrobial action,along with their applications in antimicrobials.Additionally,the synergistic mechanisms of these antimicrobial approaches are elucidated.The review provides a reference for developing multifunctional photothermal nanoplatforms for treating bacterially infected wounds.
