Study on Fatigue Cracking of Diaphragm's Arc Opening of OSD in Steel Bridges by Using Biaxial Stress Method.

Changes in loading position have a significant impact on the stress field of each vulnerable area of an orthotropic steel deck (OSD). The arc opening area of the diaphragm and the connecting area between the U-rib and the diaphragm under the moving load are prone to fatigue cracking. By comparing the stress responses under different methods, the hot spot stress (HSS) method is used as the main stress extraction method in fatigue performance evaluation. The control stress of fatigue cracking was analyzed by comparing the direction of the principal stress field with the crack direction in this experiment. According to the stress amplitude deviation under the biaxial stress state, a set of methods for evaluating the effects of in-plane biaxial fatigue was developed. An improved luffing fatigue assessment S-N curve was applied to analyze the fatigue life of the diaphragm's arc opening area. The results show that when the moving load is exactly above the connection of the deck and the web of the U-rib on one side, it is in the most unfavorable position in the transverse direction, and the diaphragm is mainly under the in-plane stress state. The longitudinal range of the stress influence line of the arc opening is approximately twice the diaphragm spacing. Two to three stress cycles are caused by one fatigue load. Fatigue crack control stress is the principal stress tangential to the arc opening's edge in this area. The normal direction of the principal stress in the model test is roughly consistent with the crack initiation direction. The variation in the stress amplitude deviation in this area is caused by changes in the action position of the moving load. When the moving load is at a certain distance from the involved diaphragm, it is reduced to zero, implying that the in-plane fatigue effect is the greatest in this area.

Experimental Analysis of the Magnetic Leakage Detection of a Corroded Steel Strand Due to Vibration.

The self-magnetic flux leakage (SMFL) detection technique has great potential in the corrosion detection of bridge stay cables due to its advantages of small testing equipment, high accuracy, and fast testing rate. However, the vibration effect in the cable's SMFL detection is unclear. To address this, the influence of vibration on the magnetic field distribution of cable structure is analyzed theoretically. According to the theoretical model, the effect of vibration on SMFL detection primarily manifests as displacement changes (displacement-added magnetic field) and defect shape changes (deformation-added magnetic field). SMFL detection experiments are conducted on steel strands. The results demonstrate that the displacement-added magnetic field exhibits statistical characteristics in the form of a normal distribution, fluctuating around the zero value. The impact of the deformation-added magnetic field on SMFL is linearly correlated with the corrosion ratio c. Moreover, a corrosion characterization index A was proposed and has an excellent linear fit with the corrosion ratio c. The index A effectively improves the accuracy of corrosion detection and provides early warning for the maintenance of cable structures.

Coexpression of Tail Fiber and Tail Protein Genes of the Cyanophage PP Using a Synthetic Genomics Approach Enhances the Salt Tolerance of Synechocystis PCC 6803.

Cyanophages are viruses that specifically infect cyanobacteria and are capable of regulating the population densities and seasonal distributions of cyanobacteria. However, few studies have investigated the interactions between cyanophages and heterologous hosts, owing to the inability of cyanophages to infect heterologous cyanobacterial hosts. Here, a truncated artificial cyanophage genome, Syn-P4-8, was designed and assembled that contained 18 genes for viral coat assembly proteins but not genes related to host infection or DNA replication. Syn-P4-8 was transferred into the heterologous host Synechocystis sp. PCC 6803 by conjugation. The growth of strain CS-02 carrying Syn-P4-8 was significantly better than that of the control strain when grown in medium containing 5% NaCl. Only two cyanophage genes, encoding the tail protein (open reading frame 25 [ORF25]) and the tail fiber protein (ORF26), were transcribed in Synechocystis PCC 6803 grown in BG11 medium supplemented with 5% NaCl. However, expression of either ORF25 or ORF26 alone could not recover this phenotype. In addition, transcriptomic analysis revealed the presence of 334 differentially expressed genes in CS-02 compared to the control strain, corresponding to 151 downregulated and 183 upregulated genes that may affect cyanobacterial salt tolerances. In this study, synthetic biology methods were used to strengthen our understanding of the interactions between cyanophage genes and heterologous hosts. IMPORTANCE We synthesized and assembled a truncated cyanophage genome called Syn-P4-8, containing 18 genes for viral coat assembly proteins, and transferred it into a nonhost strain, Synechocystis sp. PCC 6803, to investigate interactions between Syn-P4-8 and Synechocystis PCC 6803. We found that coexpression of tail fiber and tail protein genes enhanced the salt tolerance of Synechocystis PCC 6803.

Effect of PKH-26-labeled exosomes derived from bone marrow mesenchymal stem cells on corneal epithelium regeneration in diabetic mice.

Background: It is known that bone marrow mesenchymal stem cells (BM-MSCs) could speed up the regeneration of diabetic corneal epithelium. To investigate the effect of exosomes derived from mouse BM-MSCs on corneal epithelium regeneration in diabetic mice. Methods: Diabetic mouse models were established using streptozotocin (STZ), and their central corneal epithelium was scratched under a microscope. The diabetic mice were randomly divided into three groups: the control group was injected with subconjunctival phosphate buffer saline; the exosomes group was treated with a subconjunctival injection of exosomes derived from BM-MSCs; and the BM-MSCs group was treated with a subconjunctival injection of BM-MSCs. The corneal epithelium repair rates in the three groups were compared, and the distribution of the exosomes derived from BM-MSCs labeled with PKH-26 was observed by immunofluorescence. Hematoxylin-eosin staining of the corneal tissue was observed 72 h after the treatments in the three groups. Results: The diabetic mice were successfully established by a blood glucose level >16.7 mmol/L after 8 weeks. The corneal epithelium healing rates in experimental groups 1 and 2 were significantly higher than those of the control group at 24, 48, and 72 h (P<0.05). However, there was no significant difference in the corneal epithelial healing rate between experimental groups 1 and 2 (P>0.05). The exosomes derived from BM-MSCs were found in the superficial corneal stroma in experimental groups 1 and 2, with the majority of the exosomes distributed in the limbal epithelium at the edge of the injury area. The proliferation of corneal epithelial cells in experimental groups 1 and 2 was more obvious than that in the control group. Conclusions: The exosomes derived from BM-MSCs labeled with PKH-26 significantly promoted the repair of corneal epithelial injury in diabetic mice. These exosomes might be a substitute for BM-MSCs in the repair of diabetic keratopathy, suggesting a new idea for the repair of diabetic keratopathy with "cell-free" stem cell therapy, which will require a clinical study.

Numerical Simulation of Fatigue Cracking of Diaphragm Notch in Orthotropic Steel Deck Model.

Orthotropic steel deck (OSD) are widely used in steel bridges because of their many advantages, but the structures and stresses of OSD are complex and sensitive to fatigue. Based on the model test, the structural fatigue analysis of OSD is carried out by using the extended finite element method (XFEM) to understand and reveal the causes of fatigue detail cracks and the generation and propagation of fatigue cracks at the welding ends of diaphragms, U-ribs, and diaphragms, which are the main structural fatigue details of the deck. The results show that: the fatigue crack at the diaphragm opening is not caused by a single factor, but the horizontal relative displacement is the root-cause of the fatigue crack; the contribution of out-of-plane displacement to the fatigue crack is more significant than that of vertical displacement or in-plane stress, which often leads to the initiation and propagation of the fatigue crack; the crack-propagation direction is perpendicular to the contour of principal stress, and the crack propagates into the plate along the high-stress area in the horizontal direction, which is in accordance with the basic theory of crack propagation. The research methods can provide technical support for the design of similar structures.

Synthesis and assembly of full-length cyanophage A-4L genome.

Artificial cyanophages are considered to be an effective biological method to control harmful cyanobacterial bloom. However, no synthetic cyanophage genome has been constructed and where its obstacles are unclear. Here, we survey a stretch of 16 kb length sequence of cyanophage A-4L that is unclonable in Escherichia coli. We test 12 predicted promoters of cyanophage A-4L which were verified all active in E. coli. Next, we screen for eight ORFs that hindered the assembly of intermediate DNA fragments in E. coli and describe that seven ORFs in the 16 kb sequence could not be separately cloned in E. coli. All of unclonable ORFs in high-copy-number plasmid were successfully cloned using low-copy-number vector, suggesting that these ORFs were copy-number-dependent. We propose a clone strategy abandoned the promotor and the start codon that could be applied for unclonable ORFs. Last, we de novo synthesized and assembled the full-length genome of cyanophage A-4L. This work deepens the understanding of synthetic cyanophages studies.

The TelN/tos-assisted precise targeting of chromosome segments (TAPE).

INTRODUCTION: Performing genomic large segmentation experiments will promote the annotation of complex genomic functions and contribute to the synthesis of designed genomes. It is challenging to obtain and manipulate large or complex DNA sequences with high efficiency. OBJECTIVES: This study aims to develop an effective method for direct cloning of target genome sequences from different species. METHODS: The TelN/tos system and a linear plasmid vector were first used to directly clone the large genomic segments in E. coli. For the in vitro cloning reaction, two telomeric sites were developed using TelN protelomerase at the end of the linear plasmid vector. The target DNA sequence can be easily hooked with the homology arms and maintained as a linear artificial chromosome with arbitrary restriction sites in a specific E. coli strain. RESULTS: Using the linear cloning strategy, we successfully cloned the bacterial DNA fragment of 156 kb, a yeast genomic fragment of 124 kb and mammalian mitochondrial fragment of 16 kb. The results showed a considerable improvement in cloning efficiency and demonstrated the important role of vector ratio in the cloning process. CONCLUSION: Due to the high efficiency and stability, TAPE is an effective technique for DNA cloning and fundamental molecular biotechnology method in synthetic biology.

The Synthesis and Assembly of a Truncated Cyanophage Genome and Its Expression in a Heterogenous Host.

Cyanophages play an important role in regulating the dynamics of cyanobacteria communities in the hydrosphere, representing a promising biological control strategy for cyanobacterial blooms. Nevertheless, most cyanophages are host-specific, making it difficult to control blooming cyanobacteria via single or multiple cyanophages. In order to address the issue, we explore the interaction between cyanophages and their heterologous hosts, with the aim of revealing the principles of designing and constructing an artificial cyanophage genome towards multiple cyanobacterial hosts. In the present study, we use synthetic biological approaches to assess the impact of introducing a fragment of cyanophage genome into a heterologous cyanobacterium under a variety of environmental conditions. Based on a natural cyanophage A-4L genome (41,750 bp), a truncated cyanophage genome Syn-A-4-8 is synthesized and assembled in Saccharomyces cerevisiae. We found that a 351-15,930 bp area of the A-4L genome has a fragment that is lethal to Escherichia coli during the process of attempting to assemble the full-length A-4L genome. Syn-A-4-8 was successfully introduced into E. coli and then transferred into the model cyanobacterium Synechococcus elongatus PCC 7942 (Syn7942) via conjugation. Although no significant phenotypes of Syn7942 carrying Syn-A-4-8 (LS-02) could be observed under normal conditions, its growth exhibited a prolonged lag phase compared to that of the control strain under 290-millimolar NaCl stress. Finally, the mechanisms of altered salt tolerance in LS-02 were revealed through comparative transcriptomics, and ORF25 and ORF26 on Syn-A-4-8 turned out to be the key genes causing the phenotype. Our research represents an important attempt in designing artificial cyanophages towards multiple hosts, and offers new future insights into the control of cyanobacterial blooms.

Exogenous artificial DNA forms chromatin structure with active transcription in yeast.

Yeast artificial chromosomes (YACs) are important tools for sequencing, gene cloning, and transferring large quantities of genetic information. However, the structure and activity of YAC chromatin, as well as the unintended impacts of introducing foreign DNA sequences on DNA-associated biochemical events, have not been widely explored. Here, we showed that abundant genetic elements like TATA box and transcription factor-binding motifs occurred unintentionally in a previously reported data-carrying chromosome (dChr). In addition, we used state-of-the-art sequencing technologies to comprehensively profile the genetic, epigenetic, transcriptional, and proteomic characteristics of the exogenous dChr. We found that the data-carrying DNA formed active chromatin with high chromatin accessibility and H3K4 tri-methylation levels. The dChr also displayed highly pervasive transcriptional ability and transcribed hundreds of noncoding RNAs. The results demonstrated that exogenous artificial chromosomes formed chromatin structures and did not remain as naked or loose plasmids. A better understanding of the YAC chromatin nature will improve our ability to design better data-storage chromosomes.

An artificial chromosome for data storage.

DNA digital storage provides an alternative for information storage with high density and long-term stability. Here, we report the de novo design and synthesis of an artificial chromosome that encodes two pictures and a video clip. The encoding paradigm utilizing the superposition of sparsified error correction codewords and pseudo-random sequences tolerates base insertions/deletions and is well suited to error-prone nanopore sequencing for data retrieval. The entire 254 kb sequence was 95.27% occupied by encoded data. The Transformation-Associated Recombination method was used in the construction of this chromosome from DNA fragments and necessary autonomous replication sequences. The stability was demonstrated by transmitting the data-carrying chromosome to the 100th generation. This study demonstrates a data storage method using encoded artificial chromosomes via in vivo assembly for write-once and stable replication for multiple retrievals, similar to a compact disc, with potential in economically massive data distribution.

Debugging: putting the synthetic yeast chromosome to work.

Synthetic genomics aims to de novo synthesize a functional genome redesigned from natural sequences with custom features. Designed genomes provide new toolkits for better understanding organisms, evolution and the construction of cellular factories. Currently maintaining the fitness of cells with synthetic genomes is particularly challenging as defective designs and unanticipated assembly errors frequently occur. Mapping and correcting bugs that arise during the synthetic process are imperative for the successful construction of a synthetic genome that can sustain a desired cellular function. Here, we review recently developed methods used to map and fix various bugs which arise during yeast genome synthesis with the hope of providing guidance for putting the synthetic yeast chromosome to work.

Non-Destructive Testing of Steel Corrosion Fluctuation Parameters Based on Spontaneous Magnetic Flux Leakage and Its Relationship with Steel Bar Diameter.

In an actual structure, the arrangement of steel bars is complicated, there are many factors affecting the corrosion of steel bars, and these factors affect each other. However, accurately reflecting the corrosion of steel bars in actual engineering through theoretical calculations is difficult. Besides, it is impossible to detect and evaluate steel bars rust completely and accurately. This article is based on spontaneous magnetic leakage detection technology and adopts the method of stage corrosion and scanning along the reinforcing bar. Based on spontaneous magnetic flux leakage detection technology, the linear change rate of the tangential component curve of the magnetic flux leakage signal generated after the corrosion of a steel bar is studied, and a comparison is made between the steel bar coated concrete samples with different steel bar diameters. In this paper, the "origin of magnetic flux leakage signal" is defined as a reference point, which is convenient for effectively comparing the magnetic signal curves under all operating conditions. Besides, the "rust-magnetic fluctuation parameter" is proposed to accurately reflect the sudden change of leakage magnetic field caused by disconnection due to the corrosion of a steel bar. A new data processing method is provided for the non-destructive testing of steel corrosion using the spontaneous magnetic flux leakage effect, which can effectively reduce the influence of steel bar diameter on magnetic flux leakage signal and improve the precision of non-destructive testing technology of steel bar corrosion using the metal magnetic memory effect.

[DNA assembly technologies: a review].

DNA assembly is the core technology of synthetic biology. With the development of synthetic biology, researchers have developed different DNA assembly technologies that rely on DNA polymerase or DNA ligase, and also have developed some non-enzyme-dependent DNA assembly techniques to facilitate the automation of DNA assembly. The assembly of large fragments of DNA from a few hundred kb to Mb is mostly dependent on microbial recombination. In this paper, the three types of DNA assembly technologies, including enzyme-dependent, non-enzymatic and in vivo homologous recombination, are reviewed.

Cable Tension Monitoring Based on the Elasto-Magnetic Effect and the Self-Induction Phenomenon.

Cable tension monitoring is important to control the structural performance variation of cable-supported structures. Based on the elasto-magnetic effect and the self-induction phenomenon, a new non-destructive evaluation method was proposed for cable tension monitoring. The method was called the elasto-magnetic induction (EMI) method. By analyzing the working mechanism of the EMI method, a set of cable tension monitoring systems was presented. The primary coil and the induction unit of the traditional elasto-magnetic (EM) sensor were simplified into a self-induction coil. A numerical analysis was conducted to prove the validity of the EMI method. Experimental verification of the steel cable specimens was conducted to validate the feasibility of the EMI method. To process the tension monitoring, data processing and tension calculation methods were proposed. The results of the experimental verification indicated that different cables of the same batch can be calibrated by one proper equation. The results of the numerical analysis and the experimental verification demonstrated that the cable tension can be monitored both at the tension-applying stage and the tension-loss stage. The proposed EMI method and the given monitoring system are feasible to monitor the cable tension with high sensitivity, fast response, and easy installation.

Research on Internal Force Detection Method of Steel Bar in Elastic and Yielding Stage Based on Metal Magnetic Memory.

Based on the metal magnetic memory effect, this paper proposed a new non-destructive testing method for the internal tensile force detection of steel bars by analyzing the self-magnetic flux leakage (SMFL) signals. The variation of the SMFL signal of the steel bar with the tensile force indicates that the curve of the SMFL signal has a significant extreme point when the tensile force reaches about 65% of the yield tension, of which the first derivative curve has extreme points in the elastic and yielding stages, respectively. To study the variation of SMFL signal with the axial position of the steel bar under different tensile forces, a parameter reflecting the fluctuation of the SMFL signal along the steel bar is proposed. The linear relationship between this parameter and the tensile force can be used to quantitatively calculate the tensile force of steel bar. The method in this paper provides significant application prospects for the internal force detection of steel bar in the actual engineering.

Experimental Study on Residual Bending Strength of Corroded Reinforced Concrete Beam Based on Micromagnetic Sensor.

This paper presents a nondestructive test method to evaluate the residual bending strength of corroded reinforced concrete beam by analyzing the self-magnetic flux leakage (SMFL) signals. The automatic scanning device was equipped with a micromagnetic sensor and sensor-based experimental details were introduced. Next, the theoretical formula of the normal component HS(z) of the SMFL signal that originated from the corroded region was derived based on the magnetic dipole model and the experimental results were discussed. The results indicate that the experimental data of HS(z) are consistent with the theoretical calculations, both location and extent of the steel bars corrosion can be qualitatively determined by using HS(z). The gradient K of HS(z) is approximately linearly related to the loss rate, S, of the bending strength, which can be used to evaluate the residual bending strength of the corroded reinforced concrete beam. This work lays the foundation for evaluating the residual bending strength of corroded reinforced concrete beams using the SMFL signal; the micromagnetic sensor is further applied to the civil engineering.

Quantitative Study on Corrosion of Steel Strands Based on Self-Magnetic Flux Leakage.

This paper proposed a new computing method to quantitatively and non-destructively determine the corrosion of steel strands by analyzing the self-magnetic flux leakage (SMFL) signals from them. The magnetic dipole model and three growth models (Logistic model, Exponential model, and Linear model) were proposed to theoretically analyze the characteristic value of SMFL. Then, the experimental study on the corrosion detection by the magnetic sensor was carried out. The setup of the magnetic scanning device and signal collection method were also introduced. The results show that the Logistic Growth model is verified as the optimal model for calculating the magnetic field with good fitting effects. Combined with the experimental data analysis, the amplitudes of the calculated values (BxL(x,z) curves) agree with the measured values in general. This method provides significant application prospects for the evaluation of the corrosion and the residual bearing capacity of steel strand.

Study of stiffness and bearing capacity degradation of reinforced concrete beams under constant-amplitude fatigue.

For a reinforced concrete beam subjected to fatigue loads, the structural stiffness and bearing capacity will gradually undergo irreversible degeneration, leading to damage. Moreover, there is an inherent relationship between the stiffness and bearing capacity degradation and fatigue damage. In this study, a series of fatigue tests are performed to examine the degradation law of the stiffness and bearing capacity. The results pertaining to the stiffness show that the stiffness degradation of a reinforced concrete beam exhibits a very clear monotonic decreasing "S" curve, i.e., the stiffness of the beam decreases significantly at the start of the fatigue loading, it undergoes a linear decline phase in the middle for a long loading period, and before the failure, the bearing capacity decreases drastically again. The relationship between the residual stiffness and residual bearing capacity is determined based on the assumption that the residual stiffness and residual bearing capacity depend on the same damage state, and then, the bearing capacity degradation model of the reinforced concrete beam is established based on the fatigue stiffness. Through the established model and under the premise of the known residual stiffness degradation law, the degradation law of the bearing capacity is determined by using at least one residual bearing capacity test data, for which the parameters of the stiffness degradation function are considered as material constants. The results of the bearing capacity show that the bearing capacity degradation of the reinforced concrete beam also exhibits a very clear monotonic decreasing "S" curve, which is consistent with the stiffness degradation process and in good agreement with the experiment. In this study, the stiffness and bearing capacity degradation expressions are used to quantitatively describe their occurrence in reinforced concrete beams. In particular, the expression of the bearing capacity degradation can mitigate numerous destructive tests and save cost. The stiffness and bearing capacity degradation expressions for a reinforced concrete beam can be used to predict the deformation and bearing capacity of a structure during the service process and determine the structural fatigue damage and degree of degradation.

Bridge Structure Deformation Prediction Based on GNSS Data Using Kalman-ARIMA-GARCH Model.

Bridges are an essential part of the ground transportation system. Health monitoring is fundamentally important for the safety and service life of bridges. A large amount of structural information is obtained from various sensors using sensing technology, and the data processing has become a challenging issue. To improve the prediction accuracy of bridge structure deformation based on data mining and to accurately evaluate the time-varying characteristics of bridge structure performance evolution, this paper proposes a new method for bridge structure deformation prediction, which integrates the Kalman filter, autoregressive integrated moving average model (ARIMA), and generalized autoregressive conditional heteroskedasticity (GARCH). Firstly, the raw deformation data is directly pre-processed using the Kalman filter to reduce the noise. After that, the linear recursive ARIMA model is established to analyze and predict the structure deformation. Finally, the nonlinear recursive GARCH model is introduced to further improve the accuracy of the prediction. Simulation results based on measured sensor data from the Global Navigation Satellite System (GNSS) deformation monitoring system demonstrated that: (1) the Kalman filter is capable of denoising the bridge deformation monitoring data; (2) the prediction accuracy of the proposed Kalman-ARIMA-GARCH model is satisfactory, where the mean absolute error increases only from 3.402 mm to 5.847 mm with the increment of the prediction step; and (3) in comparision to the Kalman-ARIMA model, the Kalman-ARIMA-GARCH model results in superior prediction accuracy as it includes partial nonlinear characteristics (heteroscedasticity); the mean absolute error of five-step prediction using the proposed model is improved by 10.12%. This paper provides a new way for structural behavior prediction based on data processing, which can lay a foundation for the early warning of bridge health monitoring system based on sensor data using sensing technology.

[Differentiation of human amniotic epithelial cells (HAECs) into corneal epithelial cells induced by co-culture of HAECs and corneal epithelial cells in vitro].

Objective To investigate the feasibility of inducing human amniotic epithelial cells (HAECs) differentiation into corneal epithelial cells (CECs) in vitro. Methods HAECs were isolated by type II collagenase and typsin. The cells were induced by human CECs in a TranswellTM co-culture system for 2 weeks. The expressions of cytokeratin 3+12 (CK3+12), CK14, CK19, proliferating cell nuclear antigen (PCNA) proteins in the induced HAECs were detected by immunofluorescence staining; CK12, CK14, CK19, P63 mRNAs were examined by fluorescence quantitative real-time PCR (qRT-PCR); CK3+12, CK14, CK19, P63 proteins were tested by Western blotting. Results HAECs were similar to human CECs in morphology in vitro. After co-culture with CECs, the induced HAECs were positive for CK3+12, CK14, CK19 and P63. The relative expression levels of P63, CK12, CK19 and CK14 mRNAs in the induced HAECs increased 3.35, 6.76, 2.42 and 1.06 times, respectively. Conclusion HAECs can differentiate into corneal epithelial-like cells under the condition of co-culture with CECs. The differentiation potential of HAECs may make it as seed cells for tissue engineering corneal reconstruction.