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Diseases caused by pathogens severely hampered the development of aquaculture, especially largemouth bass virus (LMBV) has caused massive mortality and severe economic losses to the culture of largemouth bass (Micropterus salmoides). Considering the environmental hazards and human health, effective and environmentally friendly therapy strategy against LMBV is of vital importance and in pressing need. In the present study, a novel nanobody (NbE4) specific for LMBV was selected from a phage display nanobody library. Immunofluorescence and indirect ELISA showed that NbE4 could recognize LMBV virions and had strong binding capacity, but RT-qPCR evidenced that NBE4 did not render the virus uninfectious. Besides, antiviral drug ribavirin was used to construct a targeted drug system delivered by bacterial nanocellulose (BNC). RT-qPCR revealed that NbE4 could significantly enhance the antiviral activity of ribavirin in vitro and in vivo. The targeted drug delivery system (BNC-Ribavirin-NbE4, BRN) reduced the inflammatory response caused by LMBV infection and improved survival rate (BRN-L, 33.3 %; BRN-M, 46.7 %; BRN-H, 56.7 %)compared with control group (13.3 %), ribavirin group (RBV, 26.7 %) and BNC-ribavirin (BNC-R, 40.0 %), respectively. This research provided an effective antiviral strategy that improved the drug therapeutic effect and thus reduced the dosage.

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Streptococcosis is a highly contagious aquatic bacterial disease that poses a significant threat to tilapia. Vaccination is a well-known effective measure to prevent and control fish bacterial diseases. Among the various immunization methods, immersion vaccination is simple and can be widely used in aquaculture. Besides, nanocarrier delivery technology has been reported as an effective solution to improve the immune effect of immersion vaccine. In this study, the surface immunogenic protein (Sip) was proved to be conserved and potential to provide cross-immunoprotection for both Streptococcus agalactiae (S. agalactiae) and Streptococcus iniae (S. iniae) by multiple sequences alignment and Western blotting analysis. On this basis, we expressed and obtained the recombinant protein rSip and connected it with functionalized carbon nanotubes (CNT) to construct the nanocarrier vaccine system CNT-rSip. After immersion immunization, the immune effect of CNT-rSip against above two streptococcus infections was evaluated in tilapia based on some aspects including the serum specific antibody level, non-specific enzyme activities, immune-related genes expression and relative percent survival (RPS) after bacteria challenge. The results showed that compared with control group, CNT-rSip significantly (P < 0.05) increased the serum antibody levels, related enzyme activities including acid phosphatase, alkaline phosphatase, lysozyme and total antioxidant capacity activities, as well as the expression levels of immune-related genes from 2 to 4 weeks post immunization (wpi), and all these indexes peaked at 3 wpi. Besides, the above indexes of CNT-rSip were higher than those of rSip group with different extend during the experiment. Furthermore, the challenge test indicated that CNT-rSip provided cross-immunoprotection against S. agalactiae and S. iniae infection with RPS of 75 % and 72.41 %, respectively, which were much higher than those of other groups. Our study indicated that the nanocarrier immersion vaccine CNT-rSip could significantly improve the antibody titer and confer cross-immuneprotection against S. agalactiae and S. iniae infection in tilapia.

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Streptococcal disease has severely restricted the development of global tilapia industry, which is mainly caused by Streptococcus agalactiae (S. agalactiae) and Streptococcus iniae (S. iniae). Vaccination has been proved to be a potential strategy to control it. In this study, a multi-epitope subunit vaccine Sip-Srr (SS) was prepared based on the B-cell antigenic epitopes prediction and multiple sequence alignment analysis of Sip and Srr sequences. Furthermore, the BNC-rSS nanocarrier vaccine system was constructed by connecting the rSS protein with modified bacterial nanocellulose (BNCs) and characterized by Fourier Transform Infrared Spectroscopy and Scanning Electron Microscope, the immersion immune effect against S. agalactiae and S. iniae infection was evaluated. The results showed that compared with the control group, BNC-rSS significantly enhanced serum antibody production, related enzyme activities and immune-related genes expression. It was noteworthy that BNC-rSS vaccine improved immune protection of tilapia, with survival rates of 66.67 % (S. agalactiae) and 60.00 % (S. iniae), respectively, compared with those of rSS vaccine (30 % and 33.33 %, respectively). Our study indicated that the BNC-rSS nanovaccine could elicit robust immune responses in tilapia by immersion immunization, and had the potential to offer cross-protection against S. agalactiae and S. iniae infection in tilapia.

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Tilapia, as one of the fish widely cultured around the world, is suffering severe impact from the streptococcus disease with the deterioration of the breeding environment and the increasing of breeding density, which brings serious economic loss to tilapia farming. In this study, the surface immunogenic protein (Sip) of Streptococcus agalactiae (S. agalactiae) was selected as the potential candidate antigen and connected with bacterial nano cellulose (BNC) to construct the nanocarrier subunit vaccine (BNC-rSip), and the immersion immune effects against S. agalactiae and Streptococcus iniae (S. iniae) in Nile tilapia were evaluated on the basis of the serum antibody level, non-specific enzyme activity, the immune-related gene expression and relative percent survival (RPS). The results indicated that Sip possessed the expected immunogenicity according to the immunoinformatic analysis. Compared with the rSip group, BNC-rSip significantly induced serum antibody production and improved the innate immunity level of tilapia. After challenge, the RPS of BNC-rSip groups were 78.95% (S. agalactiae) and 67.86% (S. iniae), which were both higher than those of rSip groups,31.58% (S. agalactiae) and 35.71% (S. iniae), respectively. Our study indicated that BNC-rSip can induce protective immunity for tilapia through immersion immunization and may be an ideal candidate vaccine for controlling tilapia streptococcal disease.

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Granulomatous diseases caused by Nocardia seriously endanger the health of cultured fish. These bacteria are widely distributed, but prevention and treatment methods are very limited. Chronic granulomatous inflammation is an important pathological feature of Nocardia infection. However, the molecular mechanisms of granuloma formation and chronic inflammation are still unclear. Constructing a granuloma infection model of Nocardia is the key to exploring the pathogenesis of the disease. In this study, we established a granuloma model in the liver of largemouth bass (Micropterus salmoides) and assessed the infection process of Nocardia seriolae at different concentrations by analysing relevant pathological features. By measuring the expression of pro-inflammatory cytokines, transcription factors and a pyroptosis-related protein, we revealed the close relationship between pyroptosis and chronic inflammation of granulomas. We further analysed the immunofluorescence results and the expression of pyroptosis-related protein of macrophage infected by N. seriolae and found that N. seriolae infection induced macrophage pyroptosis in vitro. These results were proved by flow cytometry analysis of infection experiment in vivo. Our results indicated that the pyroptosis effect may be the key to inducing chronic inflammation in the fish liver and further mediating granuloma formation. In this study, we explored the molecular mechanism underlying chronic inflammation of granulomas and developed research ideas for understanding the occurrence and development of granulomatous diseases in fish.

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Viruses are the most abundant microorganisms on the earth, their existence in contaminated waters possesses a significant threat to humans. Waterborne viral infections could be fatal to sensitive population including young child, the elderly, and the immune-compromised. It is imperative to remove viruses during water treatment to better protect public health, especially in the light of evidence of detection of coronaviruses genetic fragments in raw sewage. We reported bench-scale experiments evaluating the extent and mechanisms of removal of a model virus (spring viremia of carp virus, SVCV) in water by adsorption. Microspheres made by boronic acid-modified bacterial cellulose with excellent mechanical strength were successfully fabricated as packing materials for the column to remove glycoproteins and enveloped viruses from water. The synthesized adsorbent was characterized by attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, Atomic Force Microscopy (AFM), Scanning Electron Microscopy (SEM), and Brunauer Emmett Teller (BET) measurement. The adsorption efficiency of glycoproteins was investigated by SDS-PAGE and the Broadford protein assay, while the binding capacity with the virus (spring viremia of carp virus) was monitored by cell culture to calculate the viral cytopathic effect and viral titer caused by the virus. The data obtained from the above experiments showed that ∼3-log removal of SVCV in 3 h, which significantly reduced the virus concentration from microspheres packed column. The present study provides substantial evidence to prove beyond doubt that material based on bacterial cellulose seems to have the potential for virus removal from water which can be extended to systems of significant importance.

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@#Septin is a highly conserved class of GTP-binding proteins found in eukaryotes other than higher plants.The different subtypes of Septin can form higher order structures such as filamentous or ring-like structures in the form of heterodimer to perform their functions,which can regulate physiological processes such as yeast budding and cell division,as well as participating in the defense response of the host cell.Abnormal expression or mutation of Septin is closely related to the occurrence and development of tumors and neurological diseases.This paper summarized the physiological functions of the Septin family,the effects of Septin on the occurrence and development of tumors and neurological diseases,and the role of Septin in the host immune response.

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BACKGROUND: Rice-fish symbiosis, as an ecological and green aquaculture model, is an effective measure to relieve the environmental stress from intensive aquaculture. Compared with traditional aquaculture, the altered rearing pattern and environment will make differences in muscle nutrient and quality, intestinal microbiota, body metabolism, and even disease resistance in fish. RESULTS: To investigate this, we explored the differences between rice-tilapia (aRT and bRT) and tank-tilapia (aTT and bTT) models at the periods before and after rice flowering using 16S rRNA sequencing and untargeted metabolomics. The results showed that compared with tilapia reared in the tank model, the fish body length and weight, the muscle total umami amino acid, and monounsaturated fatty acid content were obviously higher in the rice-fish model, especially after rice flowering. Compared with other groups, the intestinal microbiota diversity of fish in the bRT group was significantly higher; the dominant microbiota was Bacteroidetes and Firmicutes at the phylum level, Bacteroides and Turicibacter at the genus level, and the relative abundances of Gram-negative, potentially pathogenic, and stress-tolerant bacteria were the highest, lowest, and highest, respectively. Besides, the differential metabolite analysis indicated that rice-fish symbiosis improved the metabolic profiles and modulated the metabolic pathways in tilapia. Moreover, the correlation analysis of 16S sequencing and metabolomics showed that Bacteroides showed a positive correlation with many metabolites related to amino acid, fatty acid, and lipid metabolism. Video Abstract CONCLUSIONS: In summary, rice flowering improves the tilapia muscle nutrient, intestinal microbiota diversity, and disease resistance and modulates the host metabolism to acclimatize the comprehensive environment in rice-fish symbiosis. Specifically, rice flowering alters the microbiota abundance involved in amino acid, fatty acid, and lipid metabolism, resulting in improving the muscle nutrient and quality through the crosstalk of gut microbial and host metabolism. Our study will provide not only new insight into the gut microbiota-metabolism-phenotype axis, but also strong support for the promotion and application of rice-fish symbiosis in aquaculture.

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DNA vaccines, as an effective prophylactic technology to induce both humoral and cellular immune responses, have already been widely studied to prevent and control viral and bacterial infections in aquaculture. To find a more effective and safer way to control Micropterus salmoides rhabdovirus (MSRV) infection in largemouth bass, two different DNA vaccines expressing partial (pcDNA3.1-G2) and full-length (pcDNA3.1-G) of the MSRV G protein were developed and injected intramuscularly with different doses. The immune effect was comprehensively compared and evaluated by detecting immune-related parameters including serum antibody levels, immune-related physiological indexes, immune-related gene expression and relative survival rates in this study. The results showed that compared with the pcDNA3.1-G vaccine, the pcDNA3.1-G2 vaccine induced higher serum antibody levels, a lower nonspecific immune response in serum (ACP, SOD and T-AOC activities), higher immune-related gene expression and a higher relative survival rate. Moreover, the immune effect of pcDNA3.1-G2-vaccinated fish showed gradually higher with the increasing pcDNA3.1-G2 concentration, especially in pcDNA3.1-G2 (10µg/per fish) group, the relative survival rate reached to 82.5%, which was significant higher (p < 0.05) than pcDNA3.1-G (10µg/per fish) group. This study indicated that screening the potential core part of an antigen is an achievable strategy to improve the immunogenicity and immunoprotective effect of DNA vaccine.

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Ficus carica polysaccharides (FCPS), one of the most effective and important compo-nents in Ficus carica L., had been considered to be a beneficial immunostimulant and may be used in immunotherapy for animals and human. However, studies were little about the effect of FCPS used as immunomodulatory and the suitable dosage in fish. The present study investigated the effect of four different dietary levels of FCPS (0.1%, 0.2%, 0.4%, 0.8%) on the growth performance, innate immune responses and survival of crucian carp against Aeromonas hydrophila infection. The results showed that compared with control group, dietary FCPS had positive effects the growth performance (final weight, feed conversion ratio and survival rate) of crucian carp. FCPS induced significant higher (p < 0.05) leukocyte phagocytosis activity, serum bactericidal activity, lysozyme activity, com-plement C3, SOD activity and total protein level in the serum of crucian carp. Moreover, innate immune response of fish in FCPS groups increased first and then decreased with increasing dietary FCPS from 0.1% to 0.8%, and reached up to the peak in 0.4% dietary FCPS groups. Besides, the cumulative mortalities in FCPS groups were remarkably lower than that of control group when challenged with A. hydrophila, the relative percent survivals were 22.67%, 55.56%, 62.22% and 17.78% in 0.1% group, 0.2% group, 0.4% group and 0.8% group, respectively. These results suggested that dietary FCPS could improve the growth performance, innate immune response and disease resistance against A. hydrophila in fish, and the suitable dietary dose of FCPS was 0.4% in crucian carp.

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Viral diseases of the central nervous system (CNS) represent a major global health concern. Difficulties in treating these diseases are caused mainly by the biological tissues and barriers, which hinder the transport of drugs into the CNS. To counter this, a nanobody-mediated virus-targeting drug delivery platform (SWCNTs-P-A-Nb) is constructed for CNS viral disease therapy. Viral encephalopathy and retinopathy (VER), caused by nervous necrosis virus (NNV), is employed as a disease model. SWCNTs-P-A-Nb is successfully constructed by employing single-walled carbon nanotubes, amantadine, and NNV-specific nanobody (NNV-Nb) as the nanocarrier, anti-NNV drug, and targeting ligand, respectively. Results showed that SWCNTs-P-A-Nb has a good NNV-targeting ability in vitro and in vivo, improving the specific distribution of amantadine in NNV-infected sites under the guidance of NNV-Nb. SWCNTs-P-F-A-Nb can pass through the muscle and gill and be excreted by the kidney. SWCNTs-P-A-Nb can transport amantadine in a fast manner and prolong the action time, improving the anti-NNV activity of amantadine. Results so far have indicated that the nanobody-mediated NNV-targeting drug delivery platform is an effective method for VER therapy, providing new ideas and technologies for control of the CNS viral diseases. IMPORTANCE CNS viral diseases have resulted in many deadly epidemics throughout history and continue to pose one of the greatest threats to public health. Drug therapy remains challenging due to the complex structure and relative impermeability of the biological tissues and barriers. Therefore, development in the intelligent drug delivery platform is highly desired for CNS viral disease therapy. In the study, a nanobody-mediated virus-targeting drug delivery platform is constructed to explore the potential application of targeted therapy in CNS viral diseases. Our findings hold great promise for the application of targeted drug delivery in CNS viral disease therapy.

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The controlled actuation of liquid metal (LM) droplets has recently shown great potential in developing smart actuating systems for applications in robotics. However, there is a lack of a simple approach for the precise manipulation of multiple LM droplets in a 2D plane, which hinders the development of complex control over droplets for realizing useful robotic applications. To overcome this challenge, here, a versatile and powerful light-induced manipulation of LM droplets is presented. The key principle is to selectively activate phototransistors in an electrolyte using infrared laser beams to electrically control LM droplets via Marangoni forces. This approach shows the ability of inducing concurrent motion, splitting, and merging of multiple LM droplets simply using light without complex and bulky systems. Parameters affecting the manipulation of LM droplets are thoroughly investigated. Moreover, a vehicle carrier driven by wheels composed of multiple LM droplets for making a light-controlled relay is demonstrated. We believe such a light-induced control method for manipulating LM droplets has the potential for advancing the development of future field-programmable robotics and droplet-based soft collaborative robots.

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The spleen is a separate organ of the teleost, playing an essential role in immune reactions. The morphology of the spleen is different from the fish species. Little knowledge about the spleen structure and the blood splenic barrier (BSB) in Nile tilapia has been reported. To address this issue, we studied the histology of the spleen and the BSB in healthy Nile tilapia. The morphology of the spleen was observed, then H&E staining, modified Jame's staining, and ultrastructural techniques were performed to portion the spleen into three subregions and analyze the location of components and fibers. Thereafter, vital staining of Nile tilapia with Trypan blue was conducted to elucidate the composition and function of BSB. Histologically, the spleen could be divided into three subregions (inner, middle, and outer). The venules, clumps of lymphocytes, and vessels were separately characterized features of the outer, middle, and inner layers. Post injection, Trypan blue was intercepted in the endotheliocytes of ellipsoids in the middle layer (i.p.) or was deposited to the reticular fibers surrounding the ellipsoids (i.v.). Additionally, the amount of Trypan blue was shown to be positively correlated to that of the Acid phosphatase expressed. In conclusion, the spleen could be portioned into three subregions, and the BSB lay in the middle layer, composed of the cuboidal-shaped endotheliocytes and the surrounding reticular fibers of the ellipsoid capillaries. The present study enriched the research of immune tissues and system in tilapia and provided reference for the study of spleen in other fish species.

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To understand surface soil heavy metal pollution characteristics, and the spatial distribution and sources of pollution in the main sedimentary features of the Qaidam Basin, a total of 129 topsoil samples(0-10 cm) were collected within a 25 km radius. The concentrations of As, Ba, Cr, Mn, Nb, Ni, Pb, Ti, Zn, Zr, and ten kinds of heavy metals were determined, the degree of contamination and potential sources quantitatively analyzed based on enrichment factors(EFs), the ground accumulation index(Igeo), and the absolute principal component-multiple linear regression(APCS-MLR) receptor model. The results showed that the surface soils of the Qaidam Basin have experienced pollution from ten kinds of heavy metals since the 1960s, with varying degrees of enrichment. As and Pb represent probable point source pollutants, and the basin as a whole shows negligible to low levels of pollution. The APCS-MLR analysis showed that heavy metal pollution derives from two sources, natural factors and anthropogenic(transportation) sources. Specifically, As, Cr, Mn, Nb, Ni, Ti, Zn, and Zr are associated with natural sources, while Ba and Pb are associated with both natural and anthropogenic sources.

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Artificial muscles possess a vast potential in accelerating the development of robotics, exoskeletons, and prosthetics. Although a variety of emerging actuator technologies are reported, they suffer from several issues, such as high driving voltages, large hysteresis, and water intolerance. Here, a liquid metal artificial muscle (LMAM) is demonstrated, based on the electrochemically tunable interfacial tension of liquid metal to mimic the contraction and extension of muscles. The LMAM can work in different solutions with a wide range of pH (0-14), generating actuation strains of up to 87% at a maximum extension speed of 15 mm s-1 . More importantly, the LMAM only needs a very low driving voltage of 0.5 V. The actuating components of the LMAM are completely built from liquids, which avoids mechanical fatigue and provides actuator linkages without mechanical constraints to movement. The LMAM is used for developing several proof-of-concept applications, including controlled displays, cargo deliveries, and reconfigurable optical reflectors. The simplicity, versatility, and efficiency of the LMAM are further demonstrated by using it to actuate the caudal fin of an untethered bionic robotic fish. The presented LMAM has the potential to extend the performance space of soft actuators for applications from engineering fields to biomedical applications.

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Liquid metal has demonstrated an enormous potential for developing soft functional devices and machines. However, current liquid metal enabled machines suffer from several issues, such as the requirement of a liquid environment, generation of weak actuating forces, and insufficient maneuverability. To overcome these restrictions, here, a motor is developed based on the electrical actuation of liquid metal droplets without the need for conventional electromagnets. The approach is distinguished by (1) the encapsulation of electrolyte and multiple liquid metal droplets within an enclosed system, and (2) the creation of stable and continuous torque outside a liquid environment. In addition, a liquid metal electrical brush is introduced to operate the motor with low friction and low wear. The unique driving mechanism endows the motor with several advantages, including low friction, no sparking, low noise, versatile working environment, and being built from soft materials that could offer new opportunities for developing soft robotics.

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Iridovirus of Taiwan (TGIV) has been threatening the grouper farming since 1997, effective prophylaxis method is urgently needed. Subunit vaccine was proved to be useful to against the virus. Bath is the simplest method of vaccination and easy to be administrated without any stress to fish. In this research, we constructed a prokaryotic expression vector of TGIV's major capsid protein (MCP) to acquire the vaccine. Single-walled carbon nanotubes (SWCNTs) were used as the carrier to enhance the protective effect of bath vaccination for juvenile pearl gentian grouper (bath with concentrations of 5, 10, 20 mg/L for 6 h). Virus challenge was done after 28 days. Survival rates were calculated after 14 days. The level of antibody, activities of related enzymes in serums and expression of immune-related genes in kidneys and spleens were test. The results showed that vaccine with SWCNTs as carrier induced a higher level of antibody than that without. In addition, the activities of related enzymes (acid phosphatase, alkaline phosphatase, superoxide dismutase) and the expression of immune-related genes (Mx1, IgM, TNFαF, Lysozyme, CC chemokine 1, IL1-ß, IL-8) had a significantly increase. What's more, higher survival rates (42.10%, 77.77%, 89.47%) were provided by vaccine with SWCNTs than vaccine without SWCNTs (29.41%, 38.09%, 43.75%). This study suggests that the protective effect of vaccine that against TGIV with the method of bath vaccination could be enhanced by SWCNTs and SWCNTs could be a potential carrier for other subunit vaccines.

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Immune escape is a common feature of bacteria, viruses, parasites and even cancer cells. Our earlier work on an integrative and conjugative element (ICEr2) of Yersinia ruckeri SC09 demonstrated contributory roles of stir-1, stir-2 and stir-3 in bacterial toxicity and ability to code for immune evasion. Here, we further examined the ability of stir-4 in ICE (r2) and its encoded STIR-4 protein to mediate immune evasion using comparative genomic analysis. Additionally, the mechanisms underlying the synergistic activities of STIR-1, STIR-2, STIR-3 and STIR-4 in immune evasion were examined. Our results showed that STIR-4 did not contribute to bacterial toxicity, either in vivo nor in vitro, or show the ability to assist in bacterial immune escape. STIR-1, STIR-2, and STIR-3 formed heterotrimers in bacteria while facilitating immune evasion, which we speculate may be essential to maintain their stability. This discovery also partially explains the previous finding that a single gene can mediate immune evasion. Our data provide further knowledge on the distribution of ICE (r2)-like elements in bacteria, validating the prevalence of large-scale gene transfer in pathogens and its potential for enhancing virulence levels. Further studies are necessary to establish the biological significance of the ICE (r2) component.

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Gallium-based room-temperature liquid metals have enormous potential for realizing various applications in electronic devices, heat flow management, and soft actuators. Filling narrow spaces with a liquid metal is of great importance in rapid prototyping and circuit printing. However, it is relatively difficult to stretch or spread liquid metals into desired patterns because of their large surface tension. Here, we propose a method to fabricate a particle-based porous material which can enable the rapid and spontaneous diffusion of liquid metals within the material under a capillary force. Remarkably, such a method can allow liquid metal to diffuse along complex structures and even overcome the effect of gravity despite their large densities. We further demonstrate that the developed method can be utilized for prototyping complex three-dimensional (3D) structures via direct casting and connecting individual parts or by 3D printing. As such, we believe that the presented technique holds great promise for the development of additive manufacturing, rapid prototyping, and soft electronics using liquid metals.

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Virulent pathogenic microorganisms often enhance their infectivity through immune evasion mechanisms. Our research on the integrative and conjugative element (ICE(r2)) of the virulent fish pathogen Yersinia ruckeri SC09 led to the identification of genes related to immune evasion (designated stir-1, stir-2, stir-3 and stir-4), among which stir-1 and stir-2 were determined as the key contributors to bacterial toxicity and immune evasion. Here, we further examined the ability of stir-3 to mediate immune evasion based on detailed bioinformatic analysis of ICE(r2) from Y. ruckeri SC09. Interactions among the translated STIR-1, STIR-2, STIR-3 and STIR-4 proteins in the secretory process were additionally explored. STIR-3 was positively correlated with bacterial toxicity and inhibited host toll-like receptor (TLR) signaling by interacting with MyD88, thereby facilitating bacterial survival in host cells. Importantly, our data showed co-secretion of STIR-1, STIR-2 and STIR-3 as a complex, with secretion failure occurring in the absence of any one of these proteins. While stir-1, stir-2, stir-3 and stir-4 genes werespecific to Y. ruckeri SC09, the ICE(r2) region where these genes were located is a mobile component widely distributed in bacteria. Therefore, the potential transmission risk of these immune evasion genes requires further research attention.

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