Evaluation of transfection methods for transient gene expression in Chinese hamster ovary cells

Three transfection reagents, Lipofectamine&reg 2000, TransIT-PRO&reg and linear 25 kDa polyethylenimine were evaluated for transient expression of enhanced green fluorescent protein in Chinese hamster ovary cells. TransIT-PRO&reg was found to be more efficient under the examined conditions, but comes at an increased cost compared to the widely used PEI.

Comparison of transfection efficiency of rat dorsal root ganglion satellite glial cells with different transfection methods

In order to compare the transfection efficiency of lncRNA malat1 plasmid and lentivirus vectors in primary cultured rat DRG-derived satelite glial cells(SGCs).FISH was employed to detect the subcellular localization IncRNA-malat1l,qPCR was used to detect the expression of IncRNA-malat1 before and after transfection of the interference vector.It was found that malat1 was not silenced by IncRNA Smart silencers plasmid.qPCR results showed no significant difference in expression level of malat1 mRNA before and after transfection smart silencers plasmid of malat1 at 24 h,48h,3 d and7 d.The subcellular localization analysis found that malat1 was mainly located in the nucleus.It is speculated that it is difficult to achieve silencing of lncRNA located in the nucleus by applying interference technology of siRNA plasmid vector.qPCR results showed that the expression level of malat1 mRNA before and after malat1 transfection was significantly decreased at 24 h and 3 d compared with the normal group and the no-load group(P<0.05).In conclusion,it was suggested that the SGCs of DRG belonged to primary cells.

Effects of Different siRNA Transfection Methods on Expression of ARID4B in HepG2 Cells

[Objectives]This study aimed to optimize the siRNA transfection method for ARID4B gene and lay a foundation for further study on the effect of ARID4B low expression on the biological function of liver cancer cells HepG2.[Methods]HepG2 cells were cultured for 0,4 and 24 h,respectively and then transfected with ARID4B targeting siRNA.The mRNA expression of ARID4B gene was detected by RT-qPCR,and the protein expression was detected by Western blot.[Results]When HepG2 cells were transfected at 0 h after seeding,the expression of ARID4B was the lowest in both mRNA and protein levels,that is,the inhibition effect was the best.[Conclusions]Transfection at different time points after seeding of HepG2 cells had different inhibition effect on the expression of ARID4B.Transfection at 0 h after seeding showed the best inhibition effect.

In Vivo Transfection Methods of DNA Vaccines

Advantages and disadvantages of four transfection methods of DNA vaccines into the body were introduced in this study, including direct transfection, transfection of bacterial vectors, cationic lipofection and cationic polymer transfection. Cationic polymer had the characteristics of high transfection efficiency, simple operation, wide application, good repeatability and low cell virulence, which could be expected as a new kind of effi- cient transfection reagents. In practical application, various transfection methods have their own advantages and disadvantages, so the best choice should be made in the design process of vaccines based on targets, test cost, using objects and its convenience.

A Review on Sources,Extractions and Analysis Methods of a Sustainable Biomaterial:Tannins

Condensed and hydrolysable tannins are non-toxic natural polyphenols that are a commercial commodity industrialized for tanning hides to obtain leather and for a growing number of other industrial applications mainly to substitute petroleum-based products.They are a definite class of sustainable materials of the forestry industry.They have been in operation for hundreds of years to manufacture leather and now for a growing number of applications in a variety of other industries,such as wood adhesives,metal coating,pharmaceutical/medical applications and several others.This review presents the main sources,either already or potentially commercial of this forestry by-materials,their industrial and laboratory extraction systems,their systems of analysis with their advantages and drawbacks,be these methods so simple to even appear primitive but nonetheless of proven effectiveness,or very modern and instrumental.It constitutes a basic but essential summary of what is necessary to know of these sustainable materials.In doing so,the review highlights some of the main challenges that remain to be addressed to deliver the quality and economics of tannin supply necessary to fulfill the industrial production requirements for some materials-based uses.

A comparison study on structure-function relationship of polysaccharides obtained from sea buckthorn berries using different methods:antioxidant and bile acid-binding capacity

In this study,the structural characters,antioxidant activities and bile acid-binding ability of sea buckthorn polysaccharides(HRPs)obtained by the commonly used hot water(HRP-W),pressurized hot water(HRP-H),ultrasonic(HRP-U),acid(HRP-C)and alkali(HRP-A)assisted extraction methods were investigated.The results demonstrated that extraction methods had significant effects on extraction yield,monosaccharide composition,molecular weight,particle size,triple-helical structure,and surface morphology of HRPs except for the major linkage bands.Thermogravimetric analysis showed that HRP-U with filamentous reticular microstructure exhibited better thermal stability.The HRP-A with the lowest molecular weight and highest arabinose content possessed the best antioxidant activities.Moreover,the rheological analysis indicated that HRPs with higher galacturonic acid content and molecular weight showed higher viscosity and stronger crosslinking network(HRP-C,HRP-W and HRP-U),which exhibited stronger bile acid binding capacity.The present findings provide scientific evidence in the preparation technology of sea buckthorn polysaccharides with good antioxidant and bile acid binding capacity which are related to the structure affected by the extraction methods.

Deciphering gastric inflammation-induced tumorigenesis through multi-omics data and AI methods

Gastric cancer(GC), the fifth most common cancer globally, remains the leading cause of cancer deaths worldwide. Inflammation-induced tumorigenesis is the predominant process in GC development;therefore, systematic research in this area should improve understanding of the biological mechanisms that initiate GC development and promote cancer hallmarks. Here, we summarize biological knowledge regarding gastric inflammation-induced tumorigenesis, and characterize the multi-omics data and systems biology methods for investigating GC development. Of note, we highlight pioneering studies in multi-omics data and state-of-the-art network-based algorithms used for dissecting the features of gastric inflammation-induced tumorigenesis, and we propose translational applications in early GC warning biomarkers and precise treatment strategies. This review offers integrative insights for GC research, with the goal of paving the way to novel paradigms for GC precision oncology and prevention.

Hybrid Strategy of Partitioned and Monolithic Methods for Solving Strongly Coupled Analysis of Inverse and Direct Piezoelectric and Circuit Coupling

The inverse and direct piezoelectric and circuit coupling are widely observed in advanced electro-mechanical systems such as piezoelectric energy harvesters.Existing strongly coupled analysis methods based on direct numerical modeling for this phenomenon can be classified into partitioned or monolithic formulations.Each formulation has its advantages and disadvantages,and the choice depends on the characteristics of each coupled problem.This study proposes a new option:a coupled analysis strategy that combines the best features of the existing formulations,namely,the hybrid partitioned-monolithic method.The analysis of inverse piezoelectricity and the monolithic analysis of direct piezoelectric and circuit interaction are strongly coupled using a partitioned iterative hierarchical algorithm.In a typical benchmark problem of a piezoelectric energy harvester,this research compares the results from the proposed method to those from the conventional strongly coupled partitioned iterative method,discussing the accuracy,stability,and computational cost.The proposed hybrid concept is effective for coupled multi-physics problems,including various coupling conditions.

Stability Analysis and Performance Evaluation of Additive Mixed-Precision Runge-Kutta Methods

Additive Runge-Kutta methods designed for preserving highly accurate solutions in mixed-precision computation were previously proposed and analyzed.These specially designed methods use reduced precision for the implicit computations and full precision for the explicit computations.In this work,we analyze the stability properties of these methods and their sensitivity to the low-precision rounding errors,and demonstrate their performance in terms of accuracy and efficiency.We develop codes in FORTRAN and Julia to solve nonlinear systems of ODEs and PDEs using the mixed-precision additive Runge-Kutta(MP-ARK)methods.The convergence,accuracy,and runtime of these methods are explored.We show that for a given level of accuracy,suitably chosen MP-ARK methods may provide significant reductions in runtime.

High-Order Decoupled and Bound Preserving Local Discontinuous Galerkin Methods for a Class of Chemotaxis Models

In this paper,we explore bound preserving and high-order accurate local discontinuous Galerkin(LDG)schemes to solve a class of chemotaxis models,including the classical Keller-Segel(KS)model and two other density-dependent problems.We use the convex splitting method,the variant energy quadratization method,and the scalar auxiliary variable method coupled with the LDG method to construct first-order temporal accurate schemes based on the gradient flow structure of the models.These semi-implicit schemes are decoupled,energy stable,and can be extended to high accuracy schemes using the semi-implicit spectral deferred correction method.Many bound preserving DG discretizations are only worked on explicit time integration methods and are difficult to get high-order accuracy.To overcome these difficulties,we use the Lagrange multipliers to enforce the implicit or semi-implicit LDG schemes to satisfy the bound constraints at each time step.This bound preserving limiter results in the Karush-Kuhn-Tucker condition,which can be solved by an efficient active set semi-smooth Newton method.Various numerical experiments illustrate the high-order accuracy and the effect of bound preserving.

Review of Collocation Methods and Applications in Solving Science and Engineering Problems

The collocation method is a widely used numerical method for science and engineering problems governed by partial differential equations.This paper provides a comprehensive review of collocation methods and their applications,focused on elasticity,heat conduction,electromagnetic field analysis,and fluid dynamics.The merits of the collocation method can be attributed to the need for element mesh,simple implementation,high computational efficiency,and ease in handling irregular domain problems since the collocation method is a type of node-based numerical method.Beginning with the fundamental principles of the collocation method,the discretization process in the continuous domain is elucidated,and how the collocation method approximation solutions for solving differential equations are explained.Delving into the historical development of the collocation methods,their earliest applications and key milestones are traced,thereby demonstrating their evolution within the realm of numerical computation.The mathematical foundations of collocation methods,encompassing the selection of interpolation functions,definition of weighting functions,and derivation of integration rules,are examined in detail,emphasizing their significance in comprehending the method’s effectiveness and stability.At last,the practical application of the collocation methods in engineering contexts is emphasized,including heat conduction simulations,electromagnetic coupled field analysis,and fluid dynamics simulations.These specific case studies can underscore collocation method’s broad applicability and effectiveness in addressing complex engineering challenges.In conclusion,this paper puts forward the future development trend of the collocation method through rigorous analysis and discussion,thereby facilitating further advancements in research and practical applications within these fields.

Molecular diagnostic approaches in detecting rearranged during transfection oncogene mutations in multiple endocrine neoplasia type 2

Different types of neuroendocrine cancer,including medullary thyroid cancer(MTC)and thyroid C-cell hyperplasia,are part of multiple endocrine neoplasia type 2(MEN2).A proto-oncogene mutation of the rearranged during transfection(RET)gene changes the way that receptor tyrosine kinases work.Multiple endocrine neoplasia,a pathological condition,involves these kinases.When the RET protooncogene changes,it can cause endocrine adenomas and hyperplasia to happen at the same time or one after the other.Pheochromocytoma,medullary thyroid carcinoma,and hyperparathyroidism,alone or in combination,are present in MEN2A patients.Some patients may also have skin lichen amyloidosis or Hirschsprung's disease.Patients with MEN2A often present with MTC.MTC is aggressive and has the worst prognosis,as most patients exhibit lymph node metastasis.MTC is one of the important causes of death in patients with MEN2A.RET mutation analysis aids in identifying MEN2A symptoms and monitoring levels of calcium,thyroid hormones,calcitonin,normetanephrine,fractionated metanephrines,and parathyroid hormone.The earlier diagnosis of MTC significantly improves survival and prompts better management of MEN2A.In this editorial,we will discuss the significance of molecular diagnostic approaches in detecting RET oncogene mutations in MEN2A.

Numerical Methods for a Class of Quadratic Matrix Equations

Quadratic matrix equations arise in many elds of scienti c computing and engineering applications.In this paper,we consider a class of quadratic matrix equations.Under a certain condition,we rst prove the existence of minimal nonnegative solution for this quadratic matrix equation,and then propose some numerical methods for solving it.Convergence analysis and numerical examples are given to verify the theories and the numerical methods of this paper.

An Evidence-Based CoCoSo Framework with Double Hierarchy Linguistic Data for Viable Selection of Hydrogen Storage Methods

Hydrogen is the new age alternative energy source to combat energy demand and climate change.Storage of hydrogen is vital for a nation’s growth.Works of literature provide different methods for storing the produced hydrogen,and the rational selection of a viable method is crucial for promoting sustainability and green practices.Typically,hydrogen storage is associated with diverse sustainable and circular economy(SCE)criteria.As a result,the authors consider the situation a multi-criteria decision-making(MCDM)problem.Studies infer that previous models for hydrogen storage method(HSM)selection(i)do not consider preferences in the natural language form;(ii)weights of experts are not methodically determined;(iii)hesitation of experts during criteria weight assessment is not effectively explored;and(iv)three-stage solution of a suitable selection of HSM is unexplored.Driven by these gaps,in this paper,authors put forward a new integrated framework,which considers double hierarchy linguistic information for rating,criteria importance through inter-criteria correlation(CRITIC)for expert weight calculation,evidence-based Bayesian method for criteria weight estimation,and combined compromise solution(CoCoSo)for ranking HSMs.The applicability of the developed framework is testified by using a case example of HSM selection in India.Sensitivity and comparative analysis reveal the merits and limitations of the developed framework.

Holistic and localized preparation methods for triboelectric sensors:principles,applications and perspectives

With the arrival of intelligent terminals,triboelectric nanogenerators,as a new kind of energy converter,are considered one of the most important technologies for the next generation of intelligent electronics.As a self-powered sensor,it can greatly reduce the power consumption of the entire sensing system by transforming external mechanical energy to electricity.However,the fabrication method of triboelectric sensors largely determines their functionality and performance.This review provides an overview of various methods used to fabricate triboelectric sensors,with a focus on the processes of micro-electro-mechanical systems technology,three-dimensional printing,textile methods,template-assisted methods,and material synthesis methods for manufacturing.The working mechanisms and suitable application scenarios of various methods are outlined.Subsequently,the advantages and disadvantages of various methods are summarized,and reference schemes for the subsequent application of these methods are included.Finally,the opportunities and challenges faced by different methods are discussed,as well as their potential for application in various intelligent systems in the Internet of Things.

Bound-Preserving Discontinuous Galerkin Methods with Modified Patankar Time Integrations for Chemical Reacting Flows

In this paper,we develop bound-preserving discontinuous Galerkin(DG)methods for chemical reactive flows.There are several difficulties in constructing suitable numerical schemes.First of all,the density and internal energy are positive,and the mass fraction of each species is between 0 and 1.Second,due to the rapid reaction rate,the system may contain stiff sources,and the strong-stability-preserving explicit Runge-Kutta method may result in limited time-step sizes.To obtain physically relevant numerical approximations,we apply the bound-preserving technique to the DG methods.Though traditional positivity-preserving techniques can successfully yield positive density,internal energy,and mass fractions,they may not enforce the upper bound 1 of the mass fractions.To solve this problem,we need to(i)make sure the numerical fluxes in the equations of the mass fractions are consistent with that in the equation of the density;(ii)choose conservative time integrations,such that the summation of the mass fractions is preserved.With the above two conditions,the positive mass fractions have summation 1,and then,they are all between 0 and 1.For time discretization,we apply the modified Runge-Kutta/multi-step Patankar methods,which are explicit for the flux while implicit for the source.Such methods can handle stiff sources with relatively large time steps,preserve the positivity of the target variables,and keep the summation of the mass fractions to be 1.Finally,it is not straightforward to combine the bound-preserving DG methods and the Patankar time integrations.The positivity-preserving technique for DG methods requires positive numerical approximations at the cell interfaces,while Patankar methods can keep the positivity of the pre-selected point values of the target variables.To match the degree of freedom,we use polynomials on rectangular meshes for problems in two space dimensions.To evolve in time,we first read the polynomials at the Gaussian points.Then,suitable slope limiters can be applied to enforce the positivity of the solutions at those points,which can be preserved by the Patankar methods,leading to positive updated numerical cell averages.In addition,we use another slope limiter to get positive solutions used for the bound-preserving technique for the flux.Numerical examples are given to demonstrate the good performance of the proposed schemes.

Numerical Investigations on the Resonance Errors of Multiscale Discontinuous Galerkin Methods for One-Dimensional Stationary Schrödinger Equation

In this paper,numerical experiments are carried out to investigate the impact of penalty parameters in the numerical traces on the resonance errors of high-order multiscale discontinuous Galerkin(DG)methods(Dong et al.in J Sci Comput 66:321–345,2016;Dong and Wang in J Comput Appl Math 380:1–11,2020)for a one-dimensional stationary Schrödinger equation.Previous work showed that penalty parameters were required to be positive in error analysis,but the methods with zero penalty parameters worked fine in numerical simulations on coarse meshes.In this work,by performing extensive numerical experiments,we discover that zero penalty parameters lead to resonance errors in the multiscale DG methods,and taking positive penalty parameters can effectively reduce resonance errors and make the matrix in the global linear system have better condition numbers.

Advances in Research of Collection and Detection Methods of Microplastics in Environment

As a new type of environmental pollutants,microplastics have gradually attracted people's attention.A large number of plastics discharged into the environment by human beings are constantly aging and breaking,and finally become microplastics.Microplastics can adsorb pollutants in the environment,and their components have certain toxicity,which can cause different degrees of harm to organisms.Due to the structural characteristics of microplastic particles,such as small particle size,large specific surface area,and their distribution in different environmental media,it is very difficult to accurately detect microplastics.Reliable collection and detection methods are the key to the study of environmental behavior of microplastics.In this study,the collection and detection methods of microplastics in the environment were reviewed,and the development direction of microplastics detection technology in the future was prospected.This study has a certain reference value for the related research and the prevention and treatment of micro-plastic pollution.

Synthesis of Reviews on Auscultation, Approaches, and Methods for Engineering Structures

Topometric auscultation is used to monitor the durability of structures, measure deformations linked to the structure of a structure or to the movement of the ground over a part of the globe, set up warning systems, etc. It first appeared as a visual method and rapidly evolved through the various techniques used. Some of these techniques using topography are used in several fields (civil engineering, geodesy, topography, mechanics, nuclear engineering, hydraulics, physics, etc.). These topometric techniques have undergone major changes as a result of technological advances, growing needs in the monitoring of movements or deformations, increased requirements and new challenges. The methodology adopted depends on the measuring instrument used, the parameters to be estimated and access to the area to be measured. There are two types of methods: destructive and non-destructive. In addition to the visual method, they can also be classified as mechanical, physico-chemical, dynamometric, electrophysical and geometric. The estimated parameter varies according to the methodology adopted. It can be defined by coordinates, distances, potential, electrical resistance, etc.

Superconvergence of Direct Discontinuous Galerkin Methods:Eigen-structure Analysis Based on Fourier Approach

This paper investigates superconvergence properties of the direct discontinuous Galerkin(DDG)method with interface corrections and the symmetric DDG method for diffusion equations.We apply the Fourier analysis technique to symbolically compute eigenvalues and eigenvectors of the amplification matrices for both DDG methods with different coefficient settings in the numerical fluxes.Based on the eigen-structure analysis,we carry out error estimates of the DDG solutions,which can be decomposed into three parts:(i)dissipation errors of the physically relevant eigenvalue,which grow linearly with the time and are of order 2k for P^(k)(k=2,3)approximations;(ii)projection error from a special projection of the exact solution,which is decreasing over the time and is related to the eigenvector corresponding to the physically relevant eigenvalue;(iii)dissipative errors of non-physically relevant eigenvalues,which decay exponentially with respect to the spatial mesh sizeΔx.We observe that the errors are sensitive to the choice of the numerical flux coefficient for even degree P^(2)approximations,but are not for odd degree P^(3)approximations.Numerical experiments are provided to verify the theoretical results.