Evaluation of genetic response of mesenchymal stem cells to nanosecond pulsed electric fields by whole transcriptome sequencing

BACKGROUND Mesenchymal stem cells(MSCs)modulated by various exogenous signals have been applied extensively in regenerative medicine research.Notably,nanosecond pulsed electric fields(nsPEFs),characterized by short duration and high strength,significantly influence cell phenotypes and regulate MSCs differentiation via multiple pathways.Consequently,we used transcriptomics to study changes in messenger RNA(mRNA),long noncoding RNA(lncRNA),microRNA(miRNA),and circular RNA expression during nsPEFs application.AIM To explore gene expression profiles and potential transcriptional regulatory mechanisms in MSCs pretreated with nsPEFs.METHODS The impact of nsPEFs on the MSCs transcriptome was investigated through whole transcriptome sequencing.MSCs were pretreated with 5-pulse nsPEFs(100 ns at 10 kV/cm,1 Hz),followed by total RNA isolation.Each transcript was normalized by fragments per kilobase per million.Fold change and difference significance were applied to screen the differentially expressed genes(DEGs).Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed to elucidate gene functions,complemented by quantitative polymerase chain reaction verification.RESULTS In total,263 DEGs were discovered,with 92 upregulated and 171 downregulated.DEGs were predominantly enriched in epithelial cell proliferation,osteoblast differentiation,mesenchymal cell differentiation,nuclear division,and wound healing.Regarding cellular components,DEGs are primarily involved in condensed chromosome,chromosomal region,actin cytoskeleton,and kinetochore.From aspect of molecular functions,DEGs are mainly involved in glycosaminoglycan binding,integrin binding,nuclear steroid receptor activity,cytoskeletal motor activity,and steroid binding.Quantitative real-time polymerase chain reaction confirmed targeted transcript regulation.CONCLUSION Our systematic investigation of the wide-ranging transcriptional pattern modulated by nsPEFs revealed the differential expression of 263 mRNAs,2 miRNAs,and 65 lncRNAs.Our study demonstrates that nsPEFs may affect stem cells through several signaling pathways,which are involved in vesicular transport,calcium ion transport,cytoskeleton,and cell differentiation.

Application of a Pulsed Electric Field to Modify the Free Energy of the Active Site of an Azoreductase

The Gibbs free energy is strongly related to the stability and catalytic function of an enzyme through the energetic changes that occur in the chemical reactions the enzyme catalyzes. In this in silico study, a pulsed electric field was applied to an azoreductase, and its effect on the Gibbs free energy of molecular docking with two dyes was measured. We propose that certain stimuli from a pulsed electric field favor the structural stability of the enzyme by promoting an arrangement in the active site, potentially leading to an enhancement of enzymatic activity overall.

Phyllanthuse emblica polyphenols:Optimization of high-voltage pulsed electric field assisted extraction,an antioxidant and anti-inflammatory effects in vitro

Background:The polyphenols extraction of Phyllanthus emblica is primarily carried out using organic solvents,and assisted by physical fields such as ultrasound and microwave for extraction.High voltage pulsed electric field technology(PEF)is a non-thermal processing technology that has high efficiency and minimal damage to thermosensitive substances.PEF has been applied to plant extraction in many studies,however,the extraction of polyphenols from Phyllanthus emblica using the PEF has still not been reported;Objective:This study explores the optimal extraction process of polyphenols from Phyllanthus emblica using the PEF,and investigates its relaxation and anti-wrinkle based on anti-oxidation and anti-inflammatory experiment,in order to develop a Phyllanthus emblica extract with substantial efficacy;Materials and Methods:The method of Phyllanthus emblica extract using PEF is established,and compared with a traditional extraction method.The experimental conditions,such as electric field intensity(0.5–6.0 kV/cm),pulse times(20−120),extraction time(0–60 min)and material concentration(0.5%∼3%),are investigated and optimized using orthogonal experiments;Results:the polyphenols in the Phyllanthus emblica extract were highest at the electric field intensity of 5 kV/cm,120 pulses,extraction time of 30 min,and 2%material concentration.The PEFcontained more polyphenols than the conventional water extraction and ultrasound-assisted extraction.The Phyllanthus emblica extract had substantial antioxidant and anti-inflammatory effects,with a good clearance effect on DPPH(IC50 of 1.82%)and ABTS(IC50 of 1.80%)radicals.At the Phyllanthus emblica extract concentration of 1.25%,inflammatory factors(TNF-α)were reduced by 47.08%;and Conclusion:The PEF is a leading-edge and promising method for preparing Phyllanthus emblica extracts.

The influence of pore characteristics on rock fragmentation mechanism by high-voltage electric pulse

High-voltage electric pulse(HVEP)is an innovative low-energy and high-efficiency technique.However,the underlying physics of the electrical breakdown within the rock,and the coupling mechanism between the various physical fields involved in HVEP still need to be further understood.In this study,we establish a 2D numerical model of multi-physical field coupling of the electrical breakdown of porous rock with randomly distributed pores to investigate the effect of pore characteristics(porosity,pore media composition)on the partial electrical breakdown of rock(i.e.the generation of a plasma channel inside the rock).Our findings indicate that the generation of a plasma channel is directionally selective and extends in the direction of a weak electrical breakdown intensity.As the porosity of the rock increases,so does the intensity of the electric field in the‘electrical damage’region—the greater the porosity,the greater the effectiveness of rock-breaking.As the fraction of pore fluid(S_(water)/S_(air))gradually declines,the generation time of the plasma channel decreases,and the efficacy of rock-breaking by HVEP increases.In addition,in this study,we conducted an indoor experiment utilizing an electric pulse drill to break down the rock in order to recreate the growth mode of the plasma channel in the rock.Moreover,the experimental results are consistent with the simulation results.In addition,the development of this type of partial electrical breakdown is confirmed to be related to electrode polarity and pore characteristics via the experiment of the symmetrical needle-needle electrode arrangement,which further demonstrates the mechanism of partial electrical breakdown.This research is significant for comprehending the process of electric impulse rock-breaking and gives theoretical guidance and technological support for advancing electric impulse drilling technology.

Demulsification Behavior, Characteristics, and Performance of Surfactant Stabilized Oil-in-Water Emulsion under Bidirectional Pulsed Electric Field

As a novel electric demulsification method,bidirectional pulsed electric field(BPEF)was employed to demulsify the surfactant stabilized oil-in-water(SSO/W)emulsion for oil/water separation in this work.The demulsification behavior,characteristics,and stages under BPEF were explored.It was discovered that BPEF drove SSO/W emulsion to move and form vortexes,during which the oil droplets aggregated and accumulated to generate an oil droplet layer(ODL).ODL subsequently transformed into a continuous oil layer(COL)leading to the demulsification and separation of SSO/W emulsion.The conversion rate of ODL to COL was defined and used to evaluate the demulsification process and reflect the coalescence ability and transformation efficiency of dispersed oil droplets into COL.Furthermore,the effects of BPEF voltage,frequency,duty cycle,ratio of pulse output time,and surfactant type and content on the demulsification performance were examined.The optimal values of BPEF parameters for demulsification operation were 400 V,25 Hz,50%,and 4:1.O/W emulsion containing anionic surfactant was apt to be demulsified by BPEF,nonionic surfactant took the second place and cationic surfactant was the most difficult.A high surfactant content was not conducive to the BPEF demulsification.This work is anticipated to provide useful guidance for oil/water separation and oil recovery from actual emulsified oily wastewater by BPEF.

Migration behavior of solidification nuclei in pure Al melt under effect of electric current pulse

A mathematical model considering free nuclei was developed to reveal the migration behavior of the free nuclei. Numerical simulation results show that most of the nuclei on the top surface of the melt move downwards and distribute randomly inside the Al melt, which induces more nucleation sites resulting in grain refinement. At the same time, the effect of nuclei size on the nuclei distribution and refinement employing electric current pulse （ECP） was also investigated. The smaller nuclei migrate a short distance with the Al melt at lower speed. But for the larger nuclei, the migration downwards with higher speed benefits the refinement of interior grains of the melt. The research results help to better understand the refinement process and provide a more reasonable explanation of the grain refinement mechanism using ECP.

Effect of electrodes and thermal insulators on grain refinement by electric current pulse

The application of electric current pulse（ECP） to a solidification process refers to the immersion of electrodes into the liquid metal and the employment of thermal insulators on the upper surface of metal.In order to ascertain the effects of these two factors on the structure refinement by the ECP technique,three groups of experiments were performed with different types of electrodes or various thermal insulators.By the comparison between solidification structures under different conditions,it is followed that the electrode and the thermal insulator have an obvious influence on the grain refinement under an applied ECP,and further analysis demonstrates that the thermal conditions of the liquid surface play a vital role in the modification of solidification structure.Also,the results support the viewpoint that most of the equiaxed grains originate from the liquid surface subjected to an ECP.

Effects of electric pulse modification on liquid structure of Al-5%Cu alloy

The electric pulse modification （EP, EPM） of liquid metal is a novel method for grain refinement. The structure of EP-modified Al-5%Cu melt was characterized by high-temperature X-ray diffractometry. The results show that the Cu-containing Al clusters remarkably increase in the EP-modified melt, furthermore, these clusters in that case tend to contract due to the decrease of relevant atomic radius and the co-ordination number. This kind of liquid-phase structure leads to a more homogeneous Cu-rich phase distribution in the final solidification structure. Differential scanning calorimetry （DSC） tests indicate that the solidification super-cooling degree of the EP-modified liquid phase is 2.36 times that of the unmodified. These facts suggest that the atom cluster changes in EP-modified Al-5%Cu melt would disagree with that by EPM model previously proposed in liquid pure metal.

Heredity of Aluminum Melt Caused by Electric Pulse Modification (Ⅰ)

The heredity of aluminum melt under the action of pulse electric field was investigated by means of the remelt experiment. A new hereditary criterion under this condition was proposed; in the meantime, the differential transferability of genetic carrier in activated melt among filial generations was validated with the aid of DSC.

Effect of electric pulse treatment on solidification structure of an Al-15%Si alloy

The effect of electric pulse modifying on the solidification structure of an Al-15%Si alloy was investigated. The result shows that the primary silicon disappears sometimes and the eutectic phase is refined after the treatrnent of EP （electric pulse） though there are different modalities in different treating durations. DSC （differential scanning calorimetry） analysis indicates that the super-cooling texture decreases and the freezing range narrows evidently after the electric pulse treatment.

Recent progress in pulsed electric field ablation for liver cancer

The number of liver cancer patients is likely to continue to increase in the coming decades due to the aging of the population and changing risk factors.Traditional treatments cannot meet the needs of all patients.New treatment methods evolved from pulsed electric field ablation are expected to lead to breakthroughs in the treatment of liver cancer.This paper reviews the safety and efficacy of irreversible electroporation in clinical studies,the methods to detect and evaluate its ablation effect,the improvements in equipment and its antitumor effect,and animal and clinical trials on electrochemotherapy.We also summarize studies on the most novel nanosecond pulsed electric field ablation techniques in vitro and in vivo.These research results are certain to promote the progress of pulsed electric field in the treatment of liver cancer.

Influences of electric pulse on solidification structure of LM-29 Al-Si alloy

The metallographic structure of LM-29 aluminum-silicon alloy modified by electric pulse treatment has been investigated and compared with those untreated.The solidification structure of LM-29 alloy has been analyzed by means of M1AP3 Quantimet image processing and analysis system,and then the solidification process has been analyzed by means of differential scanning calorimetry(DSC).The results indicate that the primary silicon phase was refined remarkably by electric pulse while the tensile strength and elongation properties increased accordingly.Electric pulse treatment can also increase the binding power between silicon clusters and alloy melt matrix,as a result,the precipitation of primary silicon phase is suppressed to meet the demand of supercooling degree for nucleating,correspondingly.The electric pulse modification has great influence on the size of silicon atomic cluster as well as its distribution in the melt,subsequently,leads to the refinement of solidification structure.

Casting structure of pure aluminum by electric pulse modification at different superheated temperatures

Electric pulse modification （EPM） is a novel technique that reduces grain size by altering the structure of a melt. It was investigated that the response of the casting structure of high pure aluminum to EPM in different superheated melts. The results indicate that the grain refining effect of a given pulse electric field holds an optimal temperature range, moreover, a lower or higher superheated temperature will both disadvantage the improvements of casting structure. It essentially lies in the cooperative action between the distorted absorption of clusters and the activated capability of atoms in the aluminum melt.

Effects of electric pulse on microstructure of Al-Si alloy in liquid and solid states

In order to investigate the change in liquid microstructure of Al-Si alloytreated by electric pulse (EP), X-ray diffraction tests with liquid Al-Si alloy and ZL109 alloytreated or not by EP were carried out. The results show that the number of Al-Si atomic clustersdecreases and that of Al-Al and Si-Si atomic clusters increases for the treated samples. The testswith ZL109 alloy indicate that a large amount of primary crystal Si appears in the solidifiedmicrostructure after treated by EP. It is found that EP can change the microstructure of liquidmetal by affecting the probability of electrons appearing in different atoms (Al and Si) in theliquid metal. The combining force of different atoms decreases relatively, and that of the sameatoms increases, which is the main reason of reducing the atomic cluster with different atoms(Al-Si) and increasing the atomic cluster with the same atoms (Al-Al, Si-Si). The increasing of theatomic cluster with the same atom cluster resulted in the increasing of Si activity and the higherpoint of eutectics in the phase diagram. It makes a lot of primary silicon appeared in ZL109 alloy.

Heredity of Aluminum Melt by Electric Pulse Modification (Ⅱ)

Heredity of high pure aluminum melts under different pulse electric field was investigated by means of repetitious remelt experiment. The results indicate that the genetic coefficient by measurement of grain size of cast structure has a close relation with pulse voltage. Moreover, the hereditary law accords with the function of In = 1＋ e^-an＋β. The stability of genetic carrier （cluster） comprises in the competition between repetitious cooling and heating impulse and the effect of electric pulse modification.

Influences of pulse electric current treatment on solidification microstructures and mechanical properties of Al-Si piston alloys

Three kinds of AI-Si piston alloys were prepared and subjected to pulse electric current treatment （PECT） at different pouring temperatures. Some aspects of the solidification microstructures were examined including the morphology and the distribution of the matrix and the secondary phases by using of optical microscopy （OM）, SEM and EDS methods. Results indicate that PECT can refine the grains of α-AI in the alloys as effectively as chemical modification by sodium salt. The processing parameters of PECT on the multi-component AI-Si alloys were then optimized through the testing of tensile strength, elongation and microhardness of the prepared alloys. A new theory was put forward to explain the mechanism of PECT.

Cementite Decomposition in Spherical Graphite Iron by Electropulsing

The influence of electropulsing on cementite decomposition in the spherical graphite iron has been studied. The results indicated that the cementite was decomposed in a short time by high current density electropulsing. With increasing electropulsing time, the in situ nucleation of graphite in cementite was accompanied with the quick decomposition of cementite. The dislocation accumulation adjacent to the cementite and the quick diffusion of carbon atom by electropulsing were main reasons for the quick decomposition of cementite. The in situ nucleation of graphite in the cementite resulted from the dislocation climbing crossing the cementite lamellae.

Removal of phenol by activated alumina bed in pulsed high-voltage electric field

A new process for removing the pollutants in aqueous solution-activated alumina bed in pulsed high-voltage electric field was investigated for the removal of phenol under different conditions. The experimental results indicated the increase in removal rate with increasing applied voltage, increasing pH value of the solution, aeration, and adding Fe^2＋. The removal rate of phenol could reach 72.1% when air aeration flow rate was 1200 ml/min, and 88.2% when 0.05 mmol/L Fe^2＋ was added into the solution under the conditions of applied voltage 25 kV, initial phenol concentration of 5 mg/L, and initial pH value 5.5. The addition of sodium carbonate reduced the phenol removal rate. In the pulsed high-voltage electric field, local discharge occurred at the surface of activated alumina, which promoted phenol degradation in the thin water film. At the same time, the space-time distribution of gas-liquid phases was more uniform and the contact areas of the activated species generated from the discharge and the pollutant molecules were much wider due to the effect of the activated alumina bed. The synthetical effects of the pulsed high-voltage electric field and the activated alumina particles accelerated phenol degradation.

Electrical triggering of earthquakes:results of laboratory experiments at spring-block models

Recently published results of field and laboratory experiments on the seismic/acoustic response to injection of direct current （DC） pulses into the Earth crust or stressed rock samples raised a question on a possibility of electrical earthquake triggering. A physical mechanism of the considered phenomenon is not clear yet in view of the very low current density （10-7-10-s A/m^2） generated by the pulsed power systems at the epicenter depth （5-10 km） of local earthquakes occurred just after the current injection. The paper describes results of laboratory ＂earthquake＂ triggering by DC pulses under conditions of a spring-block model simulated the seismogenic fault. It is experimentally shown that the electric triggering of the laboratory ＂earthquake＂ （sharp slip of a movable block of the spring-block system） is possible only within a range of subcritical state of the system, when the shear stress between the movable and fixed blocks obtains 0.98-0.99 of its critical value. The threshold of electric triggering action is about 20 A/m^2 that is 7-8 orders of magnitude higher than estimated electric current density for Bishkek test site （Northern Tien Shan, Kirghizia） where the seismic response to the man-made electric action was observed. In this connection, the electric triggering phenomena may be explained by contraction of electric current in the narrow conductive areas of the faults and the corresponding increase in current density or by involving the secondary triggering mechanisms like electromagnetic stimulation of conductive fluid migration into the fault area resulted in decrease in the fault strength properties.

ATOMIC SCALE MECHANISM OF RESTRAINED EMBRITTLEMENT IN AMORPHOUS Fe_(78)B_(13)Si_9 ALLOY BY ELECTRIC PULSE RAPID ANNEALING

The direct observations of the atomic arrangements in both conventional furnace annealed and electric pulse rapid annealed Fe78B13Si9 amorphous alloy have been conducted by the lattice imaging technique in a higt resolution electron microscope. The results showed that the embrittlement of the alloy was related to the extent of atomic rearrangements during the annealing processes. The embrittlement of the alloy after 1hour conventional furnace annealing at about 270℃ is caused by the sufficient atomic rearrangements which are characterized by the growth of some bct Fe3B-like atomic short range ordering regions already existed in the as-quenched structure. Electric pulse rapid annealing can effectively retard the above-mentioned atomic rearrangements and thus restrain the embrittlement. The embrittlement only occurs when certain amount of bcc α-Fe nanocrystals are precipitated in the amorphous matrix during electric pulse rapid annealing.