- 标题
- 摘要
- 关键词
- 实验方案
- 产品
-
A strategy for preparing non-fluorescent graphene oxide quantum dots as fluorescence quenchers in quantitative real-time PCR
摘要： In recent years, graphene oxide quantum dots (GOQDs) have emerged as novel nanomaterials for optical sensing, bioimaging, clinical testing, and environmental testing. However, GOQDs demonstrate unique photoluminescence properties, with GOQDs having quantum limitations and edge effects that often affect the accuracy of the test results in the sensory field. Herein, GOQDs with a large content of hydroxyl groups and low fluorescence intensity were first prepared via an improved Fenton reaction in this study, which introduces a large amount of epoxy groups to break the C–C bonds. The synthesized GOQDs show no significant variation in the fluorescence intensity upon ultraviolet and visible light excitations. We further utilized the GOQDs as fluorescence quenchers for different fluorescent dyes in real-time fluorescence quantitative polymerase chain reaction (qRT-PCR), and verified that the addition of GOQDs (5.3 mg ml?1) into a qRT-PCR system could reduce the background fluorescence intensity of the reaction by fluorescence resonance energy transfer (FRET) during its initial stage and its non-specific amplification, and improve its specificity. In addition, the qRT-PCR method could detect two different lengths of DNA sequences with a high specificity in the 104 to 1010 copies per ml range. It is of paramount importance to carry out further investigations to establish an efficient, sensitive, and specific RT-PCR method based on the use of GOQD nanomaterials as fluorescence quenchers.
关键词： Fenton reaction,fluorescence quenchers,DNA detection,quantitative real-time PCR,graphene oxide quantum dots
更新于2025-09-23 15:19:57
-
Optimization of rDNA degradation in recombinant Hepatitis B vaccine production plant wastewater using visible light excited Ag-doped TiO2 nanophotocatalyst
摘要： As widespread distribution of recombinant DNA of genetically modified microorganisms is a threat to the environment, the aim of this research is to investigate the efficiency of photocatalytic degradation of recombinant DNA under visible light. Using response surface methodology, a comprehensive evaluation of Ag doped-TiO2 photocatalytic degradation of recombinant DNA in Hepatitis B surface antigen production plant wastewater was performed. Photocatalytic synthesis parameters including dopant content, calcination temperature, and heating rate were investigated to model and optimize the recombinant DNA degradation efficiency. The Ag doped-TiO2 nanoparticles synthesis validation was accomplished by XRD, UV-Vis diffuse reflectance spectra, FESEM and energy-dispersive X-ray spectroscopy. A quadratic polynomial equation, developed by response surface methodology, with the correlation coefficient (R2) of 0.969 ensured the good fitness of the predicted data with the experimental results. The sensitivity analysis of model indicates that the square of silver content and calcination temperature have the greatest effect on the response, while the heating rate is the least important parameter. Furthermore, the optimum conditions of Ag content of 2.1%, calcination temperature of 485 ?C, and heating rate of 8 ?C/min resulted in 80.7% rDNA degradation experimentally.
关键词： photocatalytic degradation,genetically modified microorganisms,Ag-doped TiO2,Pichia Pastoris,recombinant DNA,Real-time PCR
更新于2025-09-23 15:19:57
-
Improvement in infected wound healing in type 1 diabetic rat by the synergistic effect of photobiomodulation therapy and conditioned medium
摘要： We investigated the effects of photobiomodulation therapy (PBMT) and conditioned medium (CM) of human bone marrow mesenchymal stem cells (hBM‐MSC) individually and/or in combination on the stereological parameters and the expression of basic fibroblast growth factor (bFGF), hypoxia‐inducible factor (HIF‐1α), and stromal cell–derived factor‐1α (SDF‐1α) in a wound model infected with methicillin‐resistant Staphylococcus aureus (MRSA) in diabetic rats. CM was provided by culturing hBM‐MSCs. Type 1 diabetes mellitus (T1DM) was induced in 72 rats, divided into four groups, harboring 18 rats each: group 1 served as a control group, group 2 received PBMT, group 3 received CM, and group 4 received CM + PBMT. On days 4, 7, and 15, six animals from each group were euthanized and the skin samples were separated for stereology examination and gene expression analysis by real‐time polymerase chain reaction. In the CM + PBMT, CM, and PBMT groups, significant decreases were induced in the number of neutrophils (1460 ± 93, 1854 ± 138, 1719 ± 248) and macrophages (539 ± 69, 804 ± 63, 912 ± 41), and significant increases in the number of fibroblasts (1073 ± 116, 836 ± 75, 912 ± 41) and angiogenesis (15 230 ± 516, 13 318 ± 1116, 14 041 ± 867), compared with those of the control group (2690 ± 371, 1139 ± 145, 566 ± 90, 12 585 ± 1219). Interestingly, the findings of the stereological examination in the CM + PBMT group were statistically more significant than those in the other groups. In the PBMT group, in most cases, the expression of bFGF, HIF‐1α, and SDF‐1α, on day 4 (27.7 ± 0.14, 28.8 ± 0.52, 27.5 ± 0.54) and day 7 (26.8 ± 1.4, 29.6 ± 1.4, 28.3 ± 1.2) were more significant than those in the control (day 4, 19.3 ± 0.42, 25.5 ± 0.08, 22.6 ± 0.04; day 7, 22.3 ± 0.22, 28.3 ± 0.59, 24.3 ± 0.19) and other treatment groups. The application of PBMT + CM induced anti-inflammatory and angiogenic activities, and hastened wound healing process in a T1 DM model of MRSA infected wound.
关键词： stereology,diabetes mellitus,diabetic foot ulcer,methicillin‐resistant staphylococcal aureus,low‐level laser therapy,phobiomodulation therapy,human bone marrow mesenchymal stem cell‐conditioned medium,real‐time PCR,rat
更新于2025-09-04 15:30:14