Study on the reduction of residual stress in laser cladding layers through groove texture.

In order to develop a method for the production of crack-free cladding layers, we combined surface texturing technology with laser cladding, establishing a multi-field coupled numerical simulation model. A separate investigation was conducted into the temperature, stress, and fluid fields in laser cladding processes with and without texturing, seeking optimal cladding parameters, and conducted experiments. The results of the numerical simulations indicate that pre-set texturing effectively reduces the temperature gradient during the cladding process, thereby making the thermal cycle curve smoother. The residual stresses in the X, Y, and Z directions are reduced by 34.84%, 3.94%, and 50.22%, respectively. The introduction of texturing reduces the internal flow velocity of the melt pool, preventing the occurrence of a double vortex effect. Experimental results show that the residual stresses in the X, Y, and Z directions of the predefined textured cladding layer are reduced by approximately 41%, 8%, and 47%, respectively, compared to the non-textured cladding layer. This effectively improves the surface roughness and internal grain size of the cladding layer, with no significant defects at the metallurgical bonding positions, providing a reference for future improvements in cladding layer quality.

Macrophages in tissue repair and regeneration: insights from zebrafish.

Macrophages play crucial and versatile roles in regulating tissue repair and regeneration upon injury. However, due to their complex compositional heterogeneity and functional plasticity, deciphering the nature of different macrophage subpopulations and unraveling their dynamics and precise roles during the repair process have been challenging. With its distinct advantages, zebrafish (Danio rerio) has emerged as an invaluable model for studying macrophage development and functions, especially in tissue repair and regeneration, providing valuable insights into our understanding of macrophage biology in health and diseases. In this review, we present the current knowledge and challenges associated with the role of macrophages in tissue repair and regeneration, highlighting the significant contributions made by zebrafish studies. We discuss the unique advantages of the zebrafish model, including its genetic tools, imaging techniques, and regenerative capacities, which have greatly facilitated the investigation of macrophages in these processes. Additionally, we outline the potential of zebrafish research in addressing the remaining challenges and advancing our understanding of the intricate interplay between macrophages and tissue repair and regeneration.

Cross-organ single-cell transcriptome profiling reveals macrophage and dendritic cell heterogeneity in zebrafish.

Tissue-resident macrophages (TRMs) and dendritic cells (DCs) are highly heterogeneous and essential for immunity, tissue regeneration, and homeostasis maintenance. Here, we comprehensively profile the heterogeneity of TRMs and DCs across adult zebrafish organs via single-cell RNA sequencing. We identify two macrophage subsets: pro-inflammatory macrophages with potent phagocytosis signatures and pro-remodeling macrophages with tissue regeneration signatures in barrier tissues, liver, and heart. In parallel, one conventional dendritic cell (cDC) population with prominent antigen presentation capacity and plasmacytoid dendritic cells (pDCs) featured by anti-virus properties are also observed in these organs. Remarkably, in addition to a single macrophage/microglia population with potent phagocytosis capacity, a pDC population and two distinct cDC populations are identified in the brain. Finally, we generate specific reporter lines for in vivo tracking of macrophage and DC subsets. Our study depicts the landscape of TRMs and DCs and creates valuable tools for in-depth study of these cells in zebrafish.

Metaphocytes are IL-22BP-producing cells regulated by ETS transcription factor Spic and essential for zebrafish barrier immunity.

Metaphocytes are tissue-resident macrophage (TRM)/dendritic cell (DC)-like cells of non-hematopoietic origin in zebrafish barrier tissues. One remarkable property of metaphocytes is their ability to capture soluble antigens from the external environment via transepithelial protrusions, a unique function manifested by specialized subpopulations of the TRMs/DCs in mammal barrier tissues. Yet, how metaphocytes acquire myeloid-like cell properties from non-hematopoietic precursors and how they regulate barrier immunity remains unknown. Here, we show that metaphocytes are in situ generated from local progenitors guided by the ETS transcription factor Spic, the deficiency of which results in the absence of metaphocytes. We further document that metaphocytes are the major IL-22BP-producing cells, and the depletion of metaphocytes causes dysregulated barrier immunity that resembles the phenotype of IL-22BP-deficient mice. These findings reveal the ontogeny, development, and function of metaphocytes in zebrafish, which facilitates our understanding of the nature and function of the mammalian TRM/DC counterparts.

Numerical Simulation Study of Multi-Field Coupling for Laser Cladding of Shaft Parts.

Since shaft parts operate under harsh environments for a long time, many critical parts suffer from corrosion, wear and other problems, leading to part failure and inability to continue in service. It is imperative to repair failed parts and increase their service life. An orthogonal experimental scheme is designed to numerically simulate the process of laser cladding of Inconel 718 alloy powder on 4140 alloy structural steel based on the ANSYS simulation platform, derive the relationship equation of cladding layer thickness according to the heat balance principle, establish a finite element model, couple three modules of temperature field, stress field and fluid field, and analyze different modules to realize the monitoring of different processes of laser cladding. The optimal cladding parameters were laser power 1000 W, scanning speed 15 rad/s, spot radius 1.5 mm, thermal stress maximum value of 696 Mpa, residual stress minimum value of 281 Mpa, and the degree of influence of three factors on thermal stress maximum value: laser power > spot radius > scanning speed. The pool in the melting process appears to melt the "sharp corner" phenomenon, the internal shows a double vortex effect, with a maximum flow rate of 0.02 m/s. The solidification process shows a different shape at each stage due to the different driving forces. In this paper, multi-field-coupled numerical simulations of the laser cladding process were performed to obtain optimal cladding parameters with low residual stresses in the clad layer. The melt pool grows and expands gradually during melting, but the laser loading time is limited, and the size and shape of the melt pool are eventually fixed, and there is a vortex flowing from the center to both sides of the cross-section inside the melt pool, forming a double vortex effect. The solidification is divided into four stages to complete the transformation of the liquid phase of the melt pool to the solid phase, and the cladding layer is formed. The multi-field-coupled numerical simulation technique is used to analyze the temperature, stress and fluid fields to provide a theoretical basis for the residual stress and surface quality of the clad layer for subsequent laser cladding experiments.

Design and Simulation of a Magnetization Drive Coil Based on the Helmholtz Principle and an Experimental Study.

In order to realize the magnetization of hydrogel mixed with NdFeB powder, a magnetization drive coil based on a Helmholtz coil is designed in this paper. The 3D model of the magnetic field system is drawn by the Maxwell software 3D module, and the influence of different factors on the magnetic induction intensity is analyzed to obtain the optimized structure of the magnetization drive coil; then, the central magnetic induction intensity and magnetic induction line distribution density of the magnetization drive coil and Helmholtz coil are compared to verify the reliability of the structure optimization. The results show that the central magnetic induction intensity is the highest when the distance between the auxiliary coils is 70 mm, the central magnetic induction intensity of the magnetized drive coil is significantly higher than that of the Helmholtz coil when the number of turns is the same, and the central magnetic induction intensity of the optimized magnetized drive coil can reach 1.37 T with a more uniform and dense magnetic induction line distribution. After building the magnetization drive coil, the magnetic induction intensity of the center of the magnetization drive coil can reach 1.34 T by a handheld digital Gauss meter test, and the error is no more than 2% with the simulation result. This design approach provides a reference for the structural design and operating characteristics analysis of magnetized drive coils and shortens the design cost and development cycle of magnetized drive coils.

Modeling manufacturing resources based on manufacturability features.

Manufacturability evaluation is an effective way to shorten the development period, optimize manufacturing processes, and reduce product costs. The manufacturability of a product depends on the processing ability of specific manufacturing resources. The development of a manufacturing resources model serves as the foundation for manufacturability evaluation. To better utilize the information on manufacturing resources, this study adopted a hybrid approach by integrating the fuzzy c-means clustering algorithm and the genetic algorithm to group manufacturing resources based on manufacturing and geometric features. The information model of manufacturing resources was built by using the object-oriented method. Subsequently, the framework to evaluate manufacturing capability based on manufacturing resources was defined. Further, an application sample was exploited and its results were analyzed. The results of the subgroup showed that the hybrid algorithm was reliable and valid and helped improve the overall performance of the company chosen for this study. The proposed approach enhanced feasibility in decision-making and facilitated the management to make more informed decisions.

Application and Prospects of Hydrogel Additive Manufacturing.

Hydrogel has become a commonly used material for 3D and 4D printing due to its favorable biocompatibility and low cost. Additive manufacturing, also known as 3D printing, was originally referred to as rapid prototyping manufacturing. Variable-feature rapid prototyping technology, also known as 4D printing, is a combination of materials, mathematics, and additives. This study constitutes a literature review to address hydrogel-based additive manufacturing technologies, introducing the characteristics of commonly used 3D printing hydrogel methods, such as direct ink writing, fused deposition modeling, and stereolithography. With this review, we also investigated the stimulus types, as well as the advantages and disadvantages of various stimulus-responsive hydrogels in smart hydrogels; non-responsive hydrogels; and various applications of additive manufacturing hydrogels, such as neural catheter preparation and drug delivery. The opportunities, challenges, and future prospects of hydrogel additive manufacturing technologies are discussed.

Endoderm-Derived Myeloid-like Metaphocytes in Zebrafish Gill Mediate Soluble Antigen-Induced Immunity.

Immune cells in the mucosal barriers of vertebrates are highly heterogeneous in their origin and function. This heterogeneity is further exemplified by the recent discovery of ectoderm-derived immune cells-metaphocytes in zebrafish epidermis. Yet, whether non-hematopoiesis-derived immune cells generally exist in barrier tissues remains obscured. Here, we report the identification and characterization of an endoderm-derived immune cell population in the gill and intestine of zebrafish. Transcriptome analysis reveals that the endoderm-derived immune cells are myeloid-like cells with high similarities to the ectoderm-derived metaphocytes in epidermis. Like metaphocytes in epidermis, the endoderm-derived immune cells are non-phagocytic but professional in external soluble antigen uptake. Depletion of the endoderm-derived immune cells in gill hinder the local immune response to external soluble stimulants. This study demonstrates a general existence of non-hematopoiesis-derived immune cells in zebrafish mucosal barriers and challenges the prevalent view that resident immune cells in mucosal barriers arise exclusively from hematopoiesis.

An Ectoderm-Derived Myeloid-like Cell Population Functions as Antigen Transporters for Langerhans Cells in Zebrafish Epidermis.

Tissue-resident macrophages (TRMs) are highly heterogeneous and engage in a wide range of diverse functions. Yet, the heterogeneities of their origins and functions remain incompletely defined. Here, we report the identification and characterization of an ectoderm-derived myeloid-like cell, which we refer to as metaphocyte. We show that metaphocytes are highly similar to conventional Langerhans cells (cLCs), the resident macrophages in epidermis, in transcriptome, morphology, and anatomic location. However, unlike cLCs, metaphocytes respond neither to tissue injury nor to bacterial infection but rather sample soluble antigens from external environment through transepithelial protrusions and transfer them to cLCs via apoptosis-phagocytosis axis. This antigen transfer is critical for zebrafish to respond to soluble antigens because the depletion of metaphocytes significantly reduces cLC antigen uptake. Our study documents the existence of ectoderm-derived myeloid-like cells that manifest distinct function from conventional TRMs and opens a new paradigm for investigation of the heterogeneities of resident immune cells.

Adult zebrafish Langerhans cells arise from hematopoietic stem/progenitor cells.

The origin of Langerhans cells (LCs), which are skin epidermis-resident macrophages, remains unclear. Current lineage tracing of LCs largely relies on the promoter-Cre-LoxP system, which often gives rise to contradictory conclusions with different promoters. Thus, reinvestigation with an improved tracing method is necessary. Here, using a laser-mediated temporal-spatial resolved cell labeling method, we demonstrated that most adult LCs originated from the ventral wall of the dorsal aorta (VDA), an equivalent to the mouse aorta, gonads, and mesonephros (AGM), where both hematopoietic stem cells (HSCs) and non-HSC progenitors are generated. Further fine-fate mapping analysis revealed that the appearance of LCs in adult zebrafish was correlated with the development of HSCs, but not T cell progenitors. Finally, we showed that the appearance of tissue-resident macrophages in the brain, liver, heart, and gut of adult zebrafish was also correlated with HSCs. Thus, the results of our study challenged the EMP-origin theory for LCs.

Construction of a Gene Knockdown System Based on Catalytically Inactive ("Dead") Cas9 (dCas9) in Staphylococcus aureus.

There has been an absence of an efficient method of gene knockdown in the important human pathogen Staphylococcus aureus like RNA interference in eukaryotes. The previously developed antisense RNA technology is mainly applied for forward genetic screening but is rather limited in specific gene knockdown because of the lack of rational antisense RNA design strategies. Here we report an efficient and specific system for gene knockdown in S. aureus based on the type II clustered regularly interspaced short palindromic repeat (CRISPR) system from Streptococcus pyogenes We can achieve gene silencing with the coexpression of dCas9, an RNA-guided DNA binding protein, and a small guide RNA complementary to the target gene. With this system, we have successfully silenced diverse sets of genes varying in size and expression level in different S. aureus strains. This system exhibited high-efficiency knockdown of both essential and nonessential genes, and its effect is inducible and reversible. In addition, the system can repress the expression of multiple genes simultaneously and silence an entire operon or part of it. This RNA-guided DNA targeting system thus provides a simple, rapid, and affordable method for selective gene knockdown in S. aureusIMPORTANCEStaphylococcus aureus is an important human and animal pathogen that can cause a diversity of infectious diseases. Molecular genetic study of S. aureus has provided an avenue for the understanding of its virulence, pathogenesis, and drug resistance, leading to the discovery of new therapies for the treatment of staphylococcal infections. However, methodologies developed for genetic manipulation of S. aureus usually involve either low efficiency or laborious procedures. Here we report an RNA-guided system for gene knockdown in S. aureus and show its high efficiency and simplicity for selective gene silencing in different strains of S. aureus This simple, rapid, and affordable system may serve as a promising tool for functional gene study in S. aureus, especially for the study of essential genes, thus facilitating the understanding of this pathogen and its interaction with its hosts.

Identification of RNAIII-binding proteins in Staphylococcus aureus using tethered RNAs and streptavidin aptamers based pull-down assay.

BACKGROUND: It has been widely recognized that small RNAs (sRNAs) play important roles in physiology and virulence control in bacteria. In Staphylococcus aureus, many sRNAs have been identified and some of them have been functionally studied. Since it is difficult to identify RNA-binding proteins (RBPs), very little has been known about the RBPs in S. aureus, especially those associated with sRNAs. RESULTS: Here we adopted a tRNA scaffold streptavidin aptamer based pull-down assay to identify RBPs in S. aureus. The tethered RNA was successfully captured by the streptavidin magnetic beads, and proteins binding to RNAIII were isolated and analyzed by mass spectrometry. We have identified 81 proteins, and expressed heterologously 9 of them in Escherichia coli. The binding ability of the recombinant proteins with RNAIII was further analyzed by electrophoresis mobility shift assay, and the result indicates that proteins CshA, RNase J2, Era, Hu, WalR, Pyk, and FtsZ can bind to RNAIII. CONCLUSIONS: This study suggests that some proteins can bind to RNA III in S. aureus, and may be involved in RNA III function. And tRSA based pull-down assay is an effective method to search for RBPs in bacteria, which should facilitate the identification and functional study of RBPs in diverse bacterial species.

[Analysis on research projects for acupuncture and moxibustion supported by NSFC during the 11th 5-year-plan].

In the present paper, the authors introduce the approved and finished research projects for acupuncture and moxibustion therapies supported by the National Natural Science Foundation of China (NSFC) during the 11th 5-year-plan. A total of 194 research projects were subsidized by NSFC from 2006 to 2010. These projects include 6 aspects: meridian-collaterals, acupoint theory, acupuncture analgesia, mechanisms underlying improvement of different clinical problems, clinical trials, and moxibustion therapy. The research on acupoints has been becoming a new hotspot in recent years. Majority of the research projects focus on the mechanism of acu-moxibustion underlying improvement of different clinical problems, while fewer projects on clinical trials. During the 11th 5-year-plan, 119 projects were completed; most of them involved meridian-collateral theory, acupoint theory, mechanism of acu-moxibustion underlying improvement of clinical problems and acupuncture manipulations. Following analysis of the finished research projects, we find that 1) many neurobiological methods and techniques are commonly used in the research on meridian-collateral theory; 2) the research on acupoint theory is changing from observing the local morphological structure to identifying characteristics of the regional activated receptors, particularly under the circumstances of visceral pathological conditions and efficacies of acupoints; 3) researches on the underlying mechanism of acu-moxibusiton for improving clinical disorders mainly focused on its cerebral protective effects against cerebral ischemia, in addition, researches about other diseases on clinical conditions have been also carried out, but the related mechanisms are far from understanding; 4) In many research projects, various new methods and techniques, such as fMRI, position emission tomography, genomics, proteomics, molecular biology, nerve stem cell, etc. were widely used; 5) Randomized controlled multi-center clinical trials are definitely insufficient. Only two clinical projects were finished during the 11th 5-year-plan. Moreover, it should be noted that in the subject of acu-moxibustion, independent innovation researches are fewer; the foundation of competitive ability is relatively weaker; the supporting conditions for finishing the research projects are poorer in some institutes or districts; and the interrelations among the theoretical, experimental and clinical researches are not intimately close, leading to relative lag in the subject development.