Identification of toll-like receptor family and the immune function of new Sptlr-6 gene of Scylla paramamosain.

As a crucial member of pattern-recognition receptors (PRRs), the Tolls/Toll-like receptors (TLRs) gene family has been proven to be involved in innate immunity in crustaceans. In this study, nine members of TLR gene family were identified from the mud crab (Scylla paramamosain) transcriptome, and the structure and phylogeny of different SpTLRs were analyzed. It was found that different SpTLRs possessed three conserved structures in the TIR domain. Meanwhile, the expression patterns of different Sptlr genes in examined tissues detected by qRT-PCR had wide differences. Compared with other Sptlr genes, Sptlr-6 gene was significantly highly expressed in the hepatopancreas and less expressed in other tissues. Therefore, the function of Sptlr-6 was further investigated. The expression of the Sptlr-6 gene was up-regulated by Poly I: C, PGN stimulation and Vibrio parahaemolyticus infection. In addition, the silencing of Sptlr-6 in hepatopancreas mediated by RNAi technology resulted in the significant decrease of several conserved genes involved in innate immunity in mud crab after V. parahaemolyticus infection, including relish, myd88, dorsal, anti-lipopolysaccharide factor (ALF), anti-lipopolysaccharide factor 2 (ALF-2) and glycine-rich antimicrobial peptide (glyamp). This study provided new knowledge for the role of the Sptlr-6 gene in defense against V. parahaemolyticus infection in S. paramamosain.

Identification and function analysis of steroid hormone synthesis pathway-related gene-Hsd3b in Scylla paramamosain.

Mud crab (Scylla paramamosain) has become an important mariculture crab along the southeast coast of China due to its strong adaptability, delicious taste, and rich nutrition. Several vertebrate steroid hormones and their synthesis-related genes and receptors have been found in crustaceans, but there are few reports on their synthesis process and mechanism. 3-beta-hydroxysteroid dehydrogenase (HSD3B) is a member of the Short-chain Dehydrogenase/Reductase (SDR) family, and an indispensable protein in vertebrates' steroid hormone synthesis pathway. In this study, the SpHsd3b gene sequence was obtained from the transcriptome data of S. paramamosain, and its full-length open reading frame (ORF) was cloned. The spatial and temporal expression pattern of SpHsd3b was performed by quantitative real-time PCR (qRT-PCR). SpHsd3b dsRNA interference (RNAi) and HSD3B inhibitor (trilostane) were used to analyze the function of SpHSD3B. The results showed that the SpHsd3b gene has an 1113 bp ORF encoding 370 amino acids with a 3ß-HSD domain. SpHSD3B has lower homology with HSD3B of vertebrates and higher homology with HSD3B of crustaceans. SpHsd3b was expressed in all examined tissues in mature crabs, and its expression was significantly higher in the testes than in the ovaries. SpHsd3b expression level was highest in the middle stage of testicular development, while its expression was higher in the early and middle stages of ovarian development. RNAi experiment and trilostane injection results showed that SpHSD3B had regulatory effects on several genes related to gonadal development and steroid hormone synthesis. 15-day trilostane suppression could also inhibit ovarian development and progesterone level of hemolymph. According to the above results, crustaceans may have steroid hormone synthesis pathways like vertebrates, and the Hsd3b gene may be involved in the gonadal development of crabs. This study provides further insight into the function of genes involved in steroid hormone synthesis in crustaceans.

Nitric oxide nano-reactor DNMF/PLGA enables tumor vascular microenvironment and chemo-hyperthermia synergetic therapy.

BACKGROUND: Breast cancer ranks first among malignant tumors, of which triple-negative breast cancer (TNBC) is characterized by its highly invasive behavior and the worst prognosis. Timely diagnosis and precise treatment of TNBC are substantially challenging. Abnormal tumor vessels play a crucial role in TNBC progression and treatment. Nitric oxide (NO) regulates angiogenesis and maintains vascular homeostasis, while effective NO delivery can normalize the tumor vasculature. Accordingly, we have proposed here a tumor vascular microenvironment remodeling strategy based on NO-induced vessel normalization and extracellular matrix collagen degradation with multimodality imaging-guided nanoparticles against TNBC called DNMF/PLGA. RESULTS: Nanoparticles were synthesized using a chemotherapeutic agent doxorubicin (DOX), a NO donor L-arginine (L-Arg), ultrasmall spinel ferrites (MnFe2O4), and a poly (lactic-co-glycolic acid) (PLGA) shell. Nanoparticle distribution in the tumor was accurately monitored in real-time through highly enhanced magnetic resonance imaging and photoacoustic imaging. Near-infrared irradiation of tumor cells revealed that MnFe2O4 catalyzes the production of a large amount of reactive oxygen species (ROS) from H2O2, resulting in a cascade catalysis of L-Arg to trigger NO production in the presence of ROS. In addition, DOX activates niacinamide adenine dinucleotide phosphate oxidase to generate and supply H2O2. The generated NO improves the vascular endothelial cell integrity and pericellular contractility to promote vessel normalization and induces the activation of endogenous matrix metalloproteinases (mainly MMP-1 and MMP-2) so as to promote extravascular collagen degradation, thereby providing an auxiliary mechanism for efficient nanoparticle delivery and DOX penetration. Moreover, the chemotherapeutic effect of DOX and the photothermal effect of MnFe2O4 served as a chemo-hyperthermia synergistic therapy against TNBC. CONCLUSION: The two therapeutic mechanisms, along with an auxiliary mechanism, were perfectly combined to enhance the therapeutic effects. Briefly, multimodality image-guided nanoparticles provide a reliable strategy for the potential application in the fight against TNBC.

Transcriptomics reveals different response mechanisms of Litopenaeus vannamei hemocytes to injection of Vibrio parahaemolyticus and WSSV.

As the most important cultural crustacean species worldwide, studies about Pacific white shrimp (Litopenaeus vannamei) have received more attention. It has been well-documented that various pathogens could infect L. vannamei, resulting in huge economic losses. The studies about the responding mechanism of L. vannamei to sole pathogens such as Vibrio parahaemolyticus and white spot virus (WSSV) have been extensively reported, while the studies about the differently responding mechanisms remain unclear. In the present study, we identified the differently expressed genes (DEGs) of L. vannamei hemocytes post V. parahaemolyticus and WSSV infection with RNA-seq technology and compared the DEGs between the two groups. The results showed 2672 DEGs post the V. parahaemolyticus challenge (1079 up-regulated and 1593 down-regulated genes), while 1146 DEGs post the WSSV challenge (1067 up-regulated and 513 down-regulated genes). In addition, we screened the genes that simultaneously respond to WSSV and V. parahaemolyticus (434), solely respond to WSSV (1146), and V. parahaemolyticus challenge (2238), respectively. Six DEGs involved in innate immunity were quantified to validate the RNA-seq results, and the results confirmed the high consistency of both methods. Furthermore, we found plenty of innate immunity-related genes that responded to V. parahaemolyticus and WSSV infection, including pattern recognition receptors (PRRs), the proPO activating system, antimicrobial peptides (AMPs), and other immunity-related proteins. The results revealed that they were differently expressed after different pathogen challenges, demonstrating the complex and specific recognition systems involved in defending against the invasion of different pathogens in the environment. The present study improved our understanding of the molecular response of hemocytes of L. vannamei to V. parahaemolyticus and WSSV stimulation.

A Solid-State Electrolyte Based on Li0.95 Na0.05 FePO4 for Lithium Metal Batteries.

Solid-state electrolytes (SSEs) play a crucial role in developing lithium metal batteries (LMBs) with high safety and energy density. Exploring SSEs with excellent comprehensive performance is the key to achieving the practical application of LMBs. In this work, the great potential of Li0.95 Na0.05 FePO4 (LNFP) as an ideal SSE due to its enhanced ionic conductivity and reliable stability in contact with lithium metal anode is demonstrated. Moreover, LNFP-based composite solid electrolytes (CSEs) are prepared to further improve electronic insulation and interface stability. The CSE containing 50 wt% of LNFP (LNFP50) shows high ionic conductivity (3.58 × 10-4 S cm-1 at 25 °C) and good compatibility with Li metal anode and cathodes. Surprisingly, the LMB of Li|LNFP50|LiFePO4 cell at 0.5 C current density shows good cycling stability (151.5 mAh g-1 for 500 cycles, 96.5% capacity retention, and 99.3% Coulombic efficiency), and high-energy LMB of Li|LNFP50|Li[Ni0.8 Co0.1 Mn0.1 ]O2 cell maintains 80% capacity retention after 170 cycles, which are better than that with traditional liquid electrolytes (LEs). This investigation offers a new approach to commercializing SSEs with excellent comprehensive performance for high-performance LMBs.

A novel Dmrt gene of crustacean: Functional analysis of idmrt-2 gene in the male reproductive system from Scylla paramamosain.

The Dmrt (double-sex and mab-3 related transcription factor) gene family is considered to be a highly conserved gene family related to sex determination and sexual differentiation across species. In order to better understand the role of the idmrt-2 gene in gonad development in Scylla paramamosain, the idmrt-2 gene was cloned and analyzed. The cDNA contains a 1659 bp ORF region encoding 552 amino acids. The qRT-PCR results showed that idmrt-2 was significantly more expressed in the testis than in other tissues (p < 0.05). The expression of idmrt-2 was highest in the spermatids stage (T2 stage), followed by the mature sperms stage (T3 stage) and significantly higher than in the spermatocytes stage (T1 stage) (p < 0.05) during testicular development and the expression difference was not significant in different stages of ovarian development. RNAi studies revealed that after idmrt-2 was knocked down, the expression of Dmrt-like and foxl-2 genes in the testis decreased, as well as IAG expression in the androgenic gland. The findings suggest that idmrt-2 may be an IAG regulator and involved in testicular development.

Implementing Monocular Visual-Tactile Sensors for Robust Manipulation.

Tactile sensing is an essential capability for robots performing manipulation tasks. In this paper, we introduce a framework to build a monocular visual-tactile sensor for robotic manipulation tasks. Such a sensor is easy to manufacture with affordable ingredients and materials. Based on a marker-based detection method, the sensor can detect the contact positions on a flat or curved surface. In the case study, we have implemented a visual-tactile sensor design specifically through the framework proposed in this paper. The design is low cost and can be processed in a very short time, making it suitable for use as an exploratory study in the laboratory.