ABSTRACT
Plasmids are the most commonly used gene carriers in the field of gene synthesis and sequencing. However, the main problems faced by traditional plasmid DNA extraction technology are low extraction throughput and high production cost, so they cannot meet the growing demand. In this study, a double-magnetic-bead method (DMBM) for plasmid extraction was developed based on the principle of plasmid extraction. The effects of the input of magnetic beads, the size of plasmid DNA fragments, and the volume of bacterial on plasmid DNA extraction were explored. In addition, the quality, throughput, and cost of plasmid DNA extraction were also compared between this technique and the commercial plasmid DNA extraction kits. The results showed that the DMBM can meet the needs of extracting plasmid DNA with different cell densities and fragment lengths. Moreover, the sensitivity and quality of plasmid extraction by the DMBM method were both superior to those of the centrifugal adsorption column method. In addition, this technique could be applied on a 96-channel automated nucleic acid extractor, resulting in higher purity of the extracted plasmid DNA, 80% reduction in extraction time, and 57.1% reduction in cost. It also reduces manual operations, achieving high-throughput and low-cost plasmid DNA extraction, thus may facilitate gene synthesis and sequencing.
Subject(s)
Plasmids/genetics , DNA/genetics , Nucleic Acids , Genetic Techniques , Magnetic PhenomenaABSTRACT
Objective: To establish a nested recombinant enzyme-assisted polymerase chain reaction (RAP) technique combined with recombined mannose-binding lectin protein (M1 protein)-magnetic beads enrichment for the detection of Candida albicans (C. albicans) and Candida tropicalis (C. tropicalis) in blood samples for the early diagnosis of candidemia albicans and candidiemia tropicalis. Methods: The primer probes for highly conserved regions of the internal transcribed spacerregions of C. albicans and C. tropicalis were deigned to establish RAP assays for the detections of C. albicans and C. tropicalis; The sensitivity and reproducibility of nucleic acid tests with gradient dilutions of standard strains and specificity of nucleic acid tests with common clinical pathogens causing bloodstream infection were condcuted. M1 protein-magnetic bead enriched plasma C. albicans and C. tropicalis were used for RAP and PCR in with simulated samples and the results were compared. Results: The sensitivity of the established dual RAP assay was 2.4-2.8 copies/reaction, with higher reproducibility and specificity. M1 protein-magnetic bead enrichment of pathogen combined with the dual RAP assay could complete the detections of C. albicans and C. tropicalis in plasma within 4 hours. Fie the pathogen samples at concentration <10 CFU/ml, the number of the samples tested by RAP was higher than that tested by PCR after enrichment. Conclusion: In this study, a dual RAP assay for the detections of C. albicans and C. tropicalis in blood sample was developed, which has the advantages of accuracy, rapidity, and less contaminants and has great potential for rapid detection of Candidemia.
Subject(s)
Humans , Lectins , Candida , Candidemia , Reproducibility of Results , Polymerase Chain Reaction , Nucleic Acids , Magnetic PhenomenaABSTRACT
To explore the feasibility of applying magnetic stimulation technology to the movement control of animal robots, the influence of coil radius, number of turns and other factors on the intensity, depth and focus of magnetic stimulation was simulated and analyzed for robot pigeons. The coil design scheme was proposed. The coil was placed on the head and one of the legs of the pigeon, and the leg electromyography (EMG) was recorded when magnetic stimulation was performed. Results showed that the EMG was significantly strengthened during magnetic stimulation. With the reduction of the output frequency of the magnetic stimulation system, the output current was increased and the EMG was enhanced accordingly. Compared with the brain magnetic stimulation, sciatic nerve stimulation produced a more significant EMG enhancement response. This indicated that the magnetic stimulation system could effectively modulate the functions of brain and peripheral nerves by driving the coil. This study provides theoretical and experimental guidance for the subsequent optimization and improvement of practical coils, and lays a preliminary theoretical and experimental foundation for the implementation of magnetic stimulation motion control of animal robots.
Subject(s)
Animals , Columbidae , Robotics , Motion , Brain , Magnetic PhenomenaABSTRACT
OBJECTIVES@#To compare the effects of cell lysis method and magnetic beads method in forensic DNA identification and to explore these two methods in forensic DNA identification.@*METHODS@#The genome DNA of THP-1 cells in different quantities was extracted by the cell lysis method and magnetic beads method, and the DNA content was quantified by real-time quantitative PCR. The cell lysis method and magnetic beads method were used to type the STR of human blood with different dilution ratios.@*RESULTS@#When the numbers of THP-1 cell were 100, 400 and 800, the DNA content extracted by cell lysis method were (1.219±0.334), (5.081±0.335), (9.332±0.318) ng, respectively; and the DNA content extracted by magnetic beads method were (1.020±0.281), (3.634±0.482), (7.896±0.759) ng, respectively. When the numbers of THP-1 cells were 400 and 800, the DNA content extracted by the cell lysis method was higher than that by the magnetic beads method. The sensitivity of cell lysis method and magnetic beads method was similar in STR typing of human blood at different dilution ratios. Complete STR typing could be obtained at 100, 300 and 500-fold dilutions of blood samples, but could not be detected at 700-fold dilution. STR typing of undiluted human blood could not be detected by cell lysis method.@*CONCLUSIONS@#The cell lysis method is easy to operate and can retain template DNA to the maximum extend. It is expected to be suitable for trace blood evidence tests.
Subject(s)
Humans , Forensic Medicine , DNA/genetics , Real-Time Polymerase Chain Reaction , Magnetic Phenomena , DNA Fingerprinting/methods , Microsatellite RepeatsABSTRACT
PURPOSE@#The incidence of acute lung injury (ALI) in severe trauma patients is 48% and the mortality rate following acute respiratory distress syndrome evolved from ALI is up to 68.5%. Alveolar epithelial type 1 cells (AEC1s) and type 2 cells (AEC2s) are the key cells in the repair of injured lungs as well as fetal lung development. Therefore, the purification and culture of AEC1s and AEC2s play an important role in the research of repair and regeneration of lung tissue.@*METHODS@#Sprague-Dawley rats (3-4 weeks, 120-150 g) were purchased for experiment. Dispase and DNase I were jointly used to digest lung tissue to obtain a single-cell suspension of whole lung cells, and then magnetic bead cell sorting was performed to isolate T1α positive cells as AEC1s from the single-cell suspension by using polyclonal rabbit anti-T1a (a specific AEC1s membrane protein) antibodies combined with anti-rabbit IgG microbeads. Afterwards, alveolar epithelial cell membrane marker protein EpCAM was designed as a key label to sort AEC2s from the remaining T1α-neg cells by another positive immunomagnetic selection using monoclonal mouse anti-EpCAM antibodies and anti-mouse IgG microbeads. Cell purity was identified by immunofluorescence staining and flow cytometry.@*RESULTS@#The purity of AEC1s and AEC2s was 88.3% ± 3.8% and 92.6% ± 2.7%, respectively. The cell growth was observed as follows: AEC1s stretched within the 12-16 h, but the cells proliferated slowly; while AEC2s began to stretch after 24 h and proliferated rapidly from the 2nd day and began to differentiate after 3 days.@*CONCLUSION@#AEC1s and AEC2s sorted by this method have high purity and good viability. Therefore, our method provides a new approach for the isolation and culture of AEC1s and AEC2s as well as a new strategy for the research of lung repair and regeneration.
Subject(s)
Animals , Rats , Alveolar Epithelial Cells/cytology , Cell Culture Techniques , Cell Separation/methods , Immunoglobulin G/metabolism , Lung , Magnetic Phenomena , Rats, Sprague-DawleyABSTRACT
This study introduces a non-invasive gastrointestinal early cancer magnetic anchor ablation electrode endoscopy system, including a magnetic anchor module and ablation electrode module. The magnetic anchoring module is composed of an external guide magnet and an internal magnet, made of permanent magnet material NdFeB; the ablation electrode module is based on the modification of the front end lens hood of the existing endoscope (CMD-90D LED electronic upper gastrointestinal endoscope). The new endoscope system not only includes all the functions of the original endoscope, but also introduces magnetic anchoring to enable the ablation electrode to be accurately positioned and controllable in the process of treating tumors, avoiding the phenomenon of gastric perforation; the introduction of steep pulse electric field ablation electrodes realizes the purpose of non-invasive treatment. Its clinical application will become a new method to treat early cancer of the digestive tract.
Subject(s)
Humans , Electrodes , Endoscopes , Gastrointestinal Tract , Magnetic Phenomena , NeoplasmsABSTRACT
Electrical and magnetic brain stimulation techniques present distinct mechanisms and efficacy in the acute treatment of depression. This was an umbrella review of meta-analyses of randomized controlled trials of brain stimulation techniques for managing acute major depressive episodes. A systematic review was performed in the PubMed/MEDLINE databases from inception until March 2020. We included the English language meta-analysis with the most randomized controlled trials on the effects of any brain stimulation technique vs. control in adults with an acute depressive episode. Continuous and dichotomous outcomes were assessed. A Measurement Tool to Assess Systematic Reviews-2 was applied and the credibility of evidence was assessed using the Grading of Recommendations Assessment, Development and Evaluation framework. Seven meta-analyses were included (5,615 patients), providing evidence for different modalities of brain stimulation techniques. Three meta-analyses were evaluated as having high methodological quality, three as moderate, and one as low. The highest quality of evidence was found for high frequency-repetitive transcranial magnetic stimulation (rTMS), transcranial direct current stimulation, and bilateral rTMS. There is strong clinical research evidence to guide future clinical use of some techniques. Our results confirm the heterogeneity of the effects across these techniques, indicating that different mechanisms of action lead to different efficacy profiles.
Subject(s)
Humans , Adult , Depressive Disorder, Major/therapy , Transcranial Direct Current Stimulation , Brain , Randomized Controlled Trials as Topic , Meta-Analysis as Topic , Depression , Magnetic PhenomenaABSTRACT
As a noninvasive neuromodulation technique, transcranial magnetic stimulation (TMS) is widely used in the clinical treatment of neurological and psychiatric diseases, but the mechanism of its action is still unclear. The purpose of this paper is to investigate the effects of different frequencies of magnetic stimulation (MS) on neuronal excitability and voltage-gated potassium channels in the
Subject(s)
Animals , Mice , Action Potentials , Magnetic Phenomena , Mental Disorders , Neurons , Patch-Clamp Techniques , Potassium Channels, Voltage-GatedABSTRACT
OBJECTIVE@#In the context of coronavirus disease 2019 (COVID-19) pandemic, the subject was designed to develop a new tracheal intubation device based on magnetic navigation technology to improve the success rate of tracheal intubation and reduce the risk of occupational exposure of medical staff.@*METHODS@#The new tracheal intubation device was designed with the uniqueness of the magnetic field environment and magnetic steering of magnetic navigation technology. And preliminary magnetic navigation tracheal intubation experiments were performed on the tracheal intubation simulator.@*RESULTS@#Magnetic navigation tracheal intubation can successfully implement tracheal intubation, and the time required is lower than that of traditional laryngoscopy.@*CONCLUSIONS@#The tracheal intubation based on magnetic navigation technology is feasible, with high efficiency and easy operation. That is expected to be widely used for tracheal intubation during treatment of patients outside the hospital in the future. At the same time, magnetic navigation endotracheal intubation technology will be the key technology for the development of endotracheal intubation robots.
Subject(s)
Humans , COVID-19/therapy , Equipment Design , Feasibility Studies , Intubation, Intratracheal , Magnetic Phenomena , SARS-CoV-2 , TechnologyABSTRACT
The magnetic anchoring lung nodule positioning device is composed of a target magnet, an anchor magnet, a coaxial puncture needle and a puncture navigation template, through these, a new type of accurate positioning technology for small pulmonary nodules is derived. The device inserts the target magnet into the both sides nearby the lung nodule under the guidance of CT. Helped by the mutual attraction of the two target magnets, they can be fixed in the lung tissue, avoiding the movement in the lung, and accurately positioning the target lung nodule before surgery. In thoracoscopic surgery, the anchor magnet and the target magnet attract each other to achieve the purpose of positioning the target nodule. The device uses the characteristics of non-contact suction of magnetic materials biomedical engineering technology, eliminating the previous procedure of direct interaction with the positioning marks, finally achieves the target of precise positioning of lung nodules and rapid surgical removal.
Subject(s)
Humans , Lung , Lung Neoplasms , Magnetic Phenomena , Magnets , Solitary Pulmonary Nodule , Thoracic Surgery, Video-Assisted , Tomography, X-Ray ComputedABSTRACT
Coagulometer, known as blood coagulation analyzer, is a product that can provide accurate test results for medical diagnosis and treatment analysis by detecting a series of items closely related to thrombosis and hemostasis in coagulation reaction. On the basis of previous traditional methods, and with our deep understanding about the principles of hemagglutination detection, we propose a hemagglutination detection method by using the dual-magnetic circuit beads method. Then, the corresponding hemagglutination detection module is designed. The coagulation time of plasma can be measured by detecting the movement of the magnetic beads when the magnetic field intensity is appropriate. The activated partial thromboplastin time(APTT) of plasma is tested when the most suitable magnetic field intensity is found. The results preliminarily show that this blood coagulation test method is valid and the corresponding test module has a potential value in business.
Subject(s)
Blood Coagulation , Blood Coagulation Tests , Magnetic Phenomena , Magnetics , Partial Thromboplastin TimeABSTRACT
RESUMEN Se evaluó el uso de partículas magnéticas acopladas a proteína L para la concentración y purificación de anticuerpos monoclonales inmunoglobulina M (mIgM) contra Taenia solium. Se evaluaron tres métodos de concentración y diferentes tiempos de elución y se optimizó la proporción de partículas a la proporción de mIgM. Demostramos que: 1) con el uso partículas magnéticas no se requiere de una concentración previa de mIgM, lo que disminuye la manipulación de los anticuerpos y mejora la recuperación, 2) se puede omitir el uso de un tampón de unión, ya que el pH de la mayoría de los sobrenadantes de cultivo celular son neutros, y 3) se necesitan tiempos de elución más largos (~45 minutos) para aumentar la recuperación a un nivel mayor a 80%. El estudio demuestra que el uso de partículas magnéticas acopladas a proteína L es una herramienta simple y eficiente para la concentración y purificación de mIgM.
ABSTRACT The use of L protein coupled magnetic particles for the concentration and purification of immunoglobulin M (mIgM) monoclonal antibodies against Taenia solium was evaluated. Three concentration methods and different elution times were evaluated and the ratio of particles to the ratio of mIgM was optimized. It is demonstrated that: 1) with the use of magnetic particles, a previous concentration of mIgM is not required, which reduces the manipulation of the antibodies and improves the recovery, 2) the use of a binding buffer can be omitted, since the pH of most cell culture supernatants are neutral, and 3) longer elution times (~ 45 minutes) are needed to increase recovery to a level greater than 80%. The study demonstrates that the use of L protein-coupled magnetic particles is a simple and efficient tool for mIgM concentration and purification.
Subject(s)
Animals , Immunoglobulin M , Taenia solium , Magnetic Phenomena , Antibodies, Monoclonal , Immunoglobulin M/immunology , Taenia solium/immunology , Antibodies, Monoclonal/isolation & purificationABSTRACT
OBJECTIVE@#To observe effect of magnetic beads auricular point sticking therapy on intrapartum fever in primipara with epidural labor analgesia and explore its possible mechanism.@*METHODS@#A total of 160 primipara were randomly divided into an observation group (80 cases, 12 cases dropped off ) and a control group (80 cases, 15 cases dropped off ). The primipara in the control group received epidural labor analgesia. In the observation group, 15 min after epidural labor analgesia was performed, magnetic beads auricular point sticking therapy was given at shenmen (TF@*RESULTS@#There was no significant difference in tympanic temperature in the observation group at each time point (@*CONCLUSION@#Magnetic beads auricular point sticking therapy can reduce the amount of anesthetics, decrease the effect of epidural analgesia on primipara's body temperature regulation and labor progress, and lower the incidence of fever during labor analgesia by regulating the level of inflammatory response in the primipara.
Subject(s)
Female , Humans , Pregnancy , Analgesia, Epidural , Analgesia, Obstetrical , Analgesics , Cesarean Section , Labor, Obstetric , Magnetic PhenomenaABSTRACT
OBJECTIVE@#To construct a magnetic and catalytic hairpin assembly-based platform for detection of dual membrane proteins on exosomes.@*METHODS@#Exosomes in supernatant of breast cancer MDA-MB-231 cell culture were separated, purified and characterized. Super-resolution imaging and Western blotting were performed to confirm the expression of the membrane protein CD63 on the exosomes. Polyacrylamide gel electrophoresis was used to verify the combination of Apt-T and exosomes. Fluorescence experiments were carried out to test the feasibility of CHA nucleic acid sequence, optimize the reaction conditions, and determine the specificity of the detection platform.@*RESULTS@#Super-resolution imaging and Western blotting showed that breast cancer MDA-MB-231 cell-derived exosomes expressed abundant membrane protein CD63. Polyacrylamide gel electrophoresis indicated that Apt-T could recognize and bind to exosomes. The results of specificity test showed that the signal-to-noise ratio of the detection platform was 1.10±0.01 for detecting normal human breast epithelial cell-derived exosomes, and was 2.09±0.08 for breast cancer cell-derived exosomes.@*CONCLUSIONS@#Magnetic and catalytic hairpin assembly-based detection platform allows simultaneous detection of two membrane proteins expressed on exosomes and identification of the expressions of membrane proteins on exosomes from different sources.
Subject(s)
Humans , Blotting, Western , Exosomes , Magnetic Phenomena , Membrane Proteins , Sensitivity and SpecificityABSTRACT
<p><b>OBJECTIVE</b>In order to accurately reconstruct the acoustic source image, the application of transducer's receiving characteristics in magneto-acoustic tomography with magnetic induction (MAT-MI) is studied.</p><p><b>METHODS</b>The conductivity phantom model is built, and the magnetic acoustic signals are simulated and the acoustic sources are reconstructed according to the transducer's receiving characteristics.</p><p><b>RESULTS</b>The reconstructed image of acoustic source is consistent with the topographic shape and size of the phantom model.</p><p><b>CONCLUSION</b>MAT-MI based on the transducer's characteristics lays the foundation for further study.</p>
Subject(s)
Acoustics , Magnetic Phenomena , Magnetics , Tomography , TransducersABSTRACT
<p><b>OBJECTIVE</b>To establish a method for detecting 3 common toxigenic molds (Aspergillus, Penicillium, and Fusarium) based on non-modified magnetic beads coupled with multiple real-time PCR (NMB-multiple qPCR).</p><p><b>METHODS</b>The primers and genus-specific probes were designed based on the rDNA sequences to develop a multiple real-time PCR using non-modified magnetic bead to enrichment of fungal spores. The sensitivity, specificity and repeatability of this assay were evaluated.</p><p><b>RESULTS</b>The detection limit of this assay for spiked samples was 10(4) CFU/g, demonstrating a 10-fold greater detection sensitivity of this assay than that of real-time PCR. The NMB-multiple qPCR assay also showed good specificity and reproducibility and yielded comparable results with those by traditional colony counting method for spiked samples (P>0.05).</p><p><b>CONCLUSION</b>NMB-multiple qPCR assay we established allows rapid and sensitive detection of common mycotoxigenic fungi in paprika.</p>
Subject(s)
Aspergillus , Capsicum , Microbiology , DNA Primers , Food Contamination , Food Microbiology , Fungi , Fusarium , Magnetic Phenomena , Penicillium , Real-Time Polymerase Chain Reaction , Methods , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
This study was aimed to improve the sensitivity of magnetic induction phase shift detection system for cerebral hemorrhage. In the study, a cerebral hemorrhage model with 13 rabbits was established by injection of autologous blood and the cerebral hemorrhage was detected by utilizing magnetic induction phase shift spectroscopy (MIPSS) detection method under the feature band. Sixty five groups of phase shift spectroscopy data were obtained. According to the characteristics of cerebral hemorrhage phase shift spectroscopy under the feature hand, an effective method, B-F distribution, to diagnose the severity of cerebral hemorrhage was designed. The results showed that using MIPSS detection method under feature band, the phase shift obviously growed with increase of injection volume of autologous blood, and the phase shift induced by a 3-mL injection reached -7.750 3 degrees ± 1.420 4 degrees. B-F distribution could effectively diagnose the severity of cerebral hemorrhage. It can be concluded that the sensitivity of the cerebral hemorrhage magnetic induction detection system is improved by one order of magnitude with the MIPSS detection method under the feature band.
Subject(s)
Animals , Rabbits , Cerebral Hemorrhage , Diagnosis , Magnetic Phenomena , Magnetics , Spectrum Analysis , MethodsABSTRACT
Magnetic induction hyperthermia becomes a very important tumor treatment method at present. In order to ensure a successful operation, doctors should make hyperthermia treatment planning before surgery. Based on Integration Healthcare Enterprise (IHE) framework and Digital Imaging and Communications in Medcine (DICOM) standard, we proposed and carried out a network workflow integrated with modern medical information systems for the dissemination of information in magnetic induction hyperthermia like accurate accessing patient information and radiology image data, storing processed images, sharing and verifying hyperthermia reports. The results proved that our system could not only improve the efficiency of magnetic induction hyperthermia treatment planning, but also save medical resources and reduce labor costs.
Subject(s)
Humans , Hyperthermia, Induced , Image Processing, Computer-Assisted , Magnetic Phenomena , Neoplasms , Therapeutics , Radiology Information Systems , Systems IntegrationABSTRACT
The purpose of this paper is to report the research and design of control system of magnetic coupling centrifugal blood pump in our laboratory, and to briefly describe the structure of the magnetic coupling centrifugal blood pump and principles of the body circulation model. The performance of blood pump is not only related to materials and structure, but also depends on the control algorithm. We studied the algorithm about motor current double-loop control for brushless DC motor. In order to make the algorithm adjust parameter change in different situations, we used the self-tuning fuzzy PI control algorithm and gave the details about how to design fuzzy rules. We mainly used Matlab Simulink to simulate the motor control system to test the performance of algorithm, and briefly introduced how to implement these algorithms in hardware system. Finally, by building the platform and conducting experiments, we proved that self-tuning fuzzy PI control algorithm could greatly improve both dynamic and static performance of blood pump and make the motor speed and the blood pump flow stable and adjustable.
Subject(s)
Algorithms , Assisted Circulation , Fuzzy Logic , Heart-Assist Devices , Magnetic Phenomena , MagneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate the root length of posterior teeth after palatal expansion by magnetic expansion appliance.</p><p><b>METHODS</b>A total of 32 patients who accepted palatal expansion with magnetic expansion appliance were enrolled in the study. Pretreatment and post-treatment cone-beam computed tomography (CBCT) images were obtained. A paired t test was used to evaluate the length change, and a group t test was used to compare the tooth lengths of developing and developed teeth.</p><p><b>RESULTS</b>Anchorage teeth showed greater root resorption than non-anchorage teeth, whereas the developed posterior teeth showed reduced resorption. Moreover, the mesial buccal roots of the first molar and first premolar were reduced. No resorption was found in the developing teeth group.</p><p><b>CONCLUSION</b>The magnetic palatal expansion can reduce the root length, particularly of developed root. The developing root of anchorage teeth show greater tolerance to resorption than the developed root. Early palatal expansion treatment with magnetic expansion appliance can reduce the risk of root resorption.</p>