Novel mechanism of MicroRNA408 in callus formation from rice mature embryo.

Mature embryos are the main explants of tissue culture used in rice transgenic technology. However, the mechanism of mature embryo callus formation remains unclear. In this study, a microRNA-mediated gene regulatory network of rice calli was established using degradome sequencing. We identified a microRNA, OsmiR408, that regulates the formation of the callus derived from the mature rice embryo. OsUCLACYANIN 30 (OsUCL 30), a target gene of OsmiR408, was the most abundant cleavage mRNA in rice callus. OsUCL17 was verified as a target gene of OsmiR408 using RNA ligase-mediated 5'-RACE. In analysis of the OsmiR408 promoter reporter line and pri-miR408 transcript level, the promoter activity and transcript level of MIR408 were increased dramatically during callus formation. In phenotypic observations, OsmiR408 knockout caused severe defects in mature embryo callus formation, whereas OsmiR408 overexpression promoted callus formation. Transcriptome analysis demonstrated that OsUCLs and certain genes related to the plant hormone signal transduction and phenylpropanoid-flavonoid biosynthesis pathway had different differential expression patterns between OsmiR408 knockout and overexpression calli. Thus, OsmiR408 may regulate callus formation mainly by affecting plant hormone signal transduction and phenylpropanoid-flavonoid biosynthesis pathway. Our findings provide insight into OsmiR408/UCLs module function in callus formation.

Electroacupuncture suppresses neuronal ferroptosis to relieve chronic neuropathic pain.

Growing evidence supports the analgesic efficacy of electroacupuncture (EA) in managing chronic neuropathic pain (NP) in both patients and NP models induced by peripheral nerve injury. However, the underlying mechanisms remain incompletely understood. Ferroptosis, a novel form of programmed cell death, has been found to be activated during NP development, while EA has shown potential in promoting neurological recovery following acute cerebral injury by targeting ferroptosis. In this study, to investigate the detailed mechanism underlying EA intervention on NP, male Sprague-Dawley rats with chronic constriction injury (CCI)-induced NP model received EA treatment at acupoints ST36 and GV20 for 14 days. Results demonstrated that EA effectively attenuated CCI-induced pain hypersensitivity and mitigated neuron damage and loss in the spinal cord of NP rats. Moreover, EA reversed the oxidative stress-mediated spinal ferroptosis phenotype by upregulating reduced expression of xCT, glutathione peroxidase 4 (GPX4), ferritin heavy chain (FTH1) and superoxide dismutase (SOD) levels, and downregulating increased expression of acyl-CoA synthetase long-chain family member 4 (ACSL4), malondialdehyde levels and iron overload. Furthermore, EA increased the immunofluorescence co-staining of GPX4 in neurons cells of the spinal cord of CCI rats. Mechanistic analysis unveiled that the inhibition of antioxidant pathway of Nrf2 signalling via its specific inhibitor, ML385, significantly countered EA's protective effect against neuronal ferroptosis in NP rats while marginally diminishing its analgesic effect. These findings suggest that EA treatment at acupoints ST36 and GV20 may protect against NP by inhibiting neuronal ferroptosis in the spinal cord, partially through the activation of Nrf2 signalling.

Angstrom-Scale 2D Channels Designed For Osmotic Energy Harvesting.

Confronting the impending exhaustion of traditional energy, it is urgent to devise and deploy sustainable clean energy alternatives. Osmotic energy contained in the salinity gradient of the sea-river interface is an innovative, abundant, clean, and renewable osmotic energy that has garnered considerable attention in recent years. Inspired by the impressively intelligent ion channels in nature, the developed angstrom-scale 2D channels with simple fabrication process, outstanding design flexibility, and substantial charge density exhibit excellent energy conversion performance, opening up a new era for osmotic energy harvesting. However, this attractive research field remains fraught with numerous challenges, particularly due to the complexities associated with the regulation at angstrom scale. In this review, the latest advancements in the design of angstrom-scale 2D channels are primarily outlined for harvesting osmotic energy. Drawing upon the analytical framework of osmotic power generation mechanisms and the insights gleaned from the biomimetic intelligent devices, the design strategies are highlighted for high-performance angstrom channels in terms of structure, functionalization, and application, with a particular emphasis on ion selectivity and ion transport resistance. Finally, current challenges and future prospects are discussed to anticipate the emergence of more anomalous properties and disruptive technologies that can promote large-scale power generation.

UBE2T mediates SORBS3 ubiquitination to enhance IL-6/STAT3 signaling and promote lung adenocarcinoma progression.

UBE2T is an oncogene in varying tumors, including lung adenocarcinoma (LUAD). SORBS3 is an important signaling regulatory protein that plays a crucial role in many cancers. This study aimed to investigate whether UBE2T promoted LUAD development by mediating the ubiquitination of SORBS3 and further explore its mechanism. Bioinformatics analysis was conducted to examine the expression of SORBS3 in LUAD tissues. Cell Counting Kit-8, Transwell, and flow cytometry were employed to analyze the cellular functions of SORBS3. Co-immunoprecipitation and ubiquitination analysis were employed to observe the correlation between UBE2T and SORBS3. In vitro and in vivo experiments verified the role of UBE2T in mediating SORBS3 ubiquitination to enhance interleukin-6/signal transducer and activator of transcription 3 (IL-6/STAT3) signaling and promote LUAD development. We observed significant downregulation of SORBS3 in LUAD tissues and cells. Furthermore, SORBS3 inhibited the proliferation, migration, and invasion of LUAD cells, while facilitating apoptosis in vitro. UBE2T enhanced IL-6/STAT3 signaling by mediating ubiquitination and degradation of SORBS3, thereby promoting LUAD progression. Additionally, this mechanism was further validated in the xenograft animal model in vivo. This study confirmed that UBE2T-mediated SORBS3 ubiquitination enhanced IL-6/STAT3 signaling and promoted LUAD progression, providing a novel therapeutic target for LUAD.

Comparing anticoagulant therapy alone, anticoagulant therapy in combination with catheter-directed thrombolysis, and anticoagulant therapy in combination with pharmacomechanical catheter-directed thrombolysis in the patients with optional inferior vena cava filter-related thrombosis: A single-center retrospective study.

OBJECTIVE: To analyze the treatment methods and efficacy of inferior vena cava filter thrombosis (IVCFT). MATERIALS AND METHODS: In this retrospective study, the clinical data for 47 patients with IVCFT who underwent sequential treatment at the Department of Vascular Surgery, Tianjin Medical University Second Hospital, from January 2020 to January 2023 were analyzed. Patients were divided into three groups according to the treatment method: anticoagulant therapy (AC group), anticoagulation plus catheter-directed thrombolysis (CDT group), and anticoagulation plus AngioJet thrombectomy plus catheter-directed thrombolysis (PCDT group). The evaluation criteria for efficacy mainly included preoperative and postoperative clinical symptoms (Villalta score), thrombus diameter, thrombus clearance rate, filter retrieval rate, filter retention time, and urokinase dosage. RESULTS: This study included 47 patients, of whom 31 were males (65.9%) and 16 females (34.1%), with a mean age of 72.05 ± 8.32 years. An Aegisy filter was used in seven patients, whereas an Illicium filter was used in forty patients. There were a total of nineteen patients in the anticoagulation-only group, with complete dissolution of the intraluminal thrombus in five patients, a residual thrombus with a maximum diameter ≤1 cm in three patients, and a residual thrombus with a maximum diameter >1 cm in eleven patients. The Villalta score was 7.16 ± 0.6 before treatment and decreased to 3.79 ± 0.59 after treatment. The thrombus diameter decreased from an average of 1.46 ± 0.2 cm before treatment to an average of 0.85 ± 0.14 cm after treatment. The retrieval rate for the filters was 42.11% (8/19), with an average dwell time of 27.4 ± 1.3 days for the filters. The CDT group consisted of 17 patients. Among whom we observed, complete dissolution of the intraluminal thrombus was observed in six patients, residual thrombus with a maximum diameter ≤1 cm in nine patients, and residual thrombus with a maximum diameter >1 cm in two patients. The Villalta score decreased from 7.53 ± 0.83 before treatment to 2.06 ± 0.39 after treatment. The thrombus diameter also decreased from 1.46 ± 0.16 cm before treatment to 0.35 ± 0.11 cm after treatment. The retrieval rate of the filters was 88.24% (15/17), and the average filter indwelling time was 19.25 ± 4.5 days. The PCDT group consisted of 11 patients. We observed complete dissolution of the intraluminal thrombus in four patients, residual thrombus with a maximum diameter ≤1 cm in six patients, and residual thrombus with a maximum diameter >1 cm in one patient. The Villalta score decreased from 7.45 ± 0.76 before treatment to 2.09 ± 0.55 after treatment. The thrombus diameter decreased from 1.50 ± 0.21 cm before treatment to 0.33 ± 0.35 cm after treatment, and the rate of filter retrieval was 90.91% (10/11). CONCLUSION: The three treatments of anticoagulation therapy, CDT, and PCDT were meaningful for preoperative and postoperative thrombolysis and symptom improvement in patients with IVCFT. The application of CDT and PCDT was superior to anticoagulation therapy, while there was no significant difference between the CDT and PCDT group. The retrieval rate of filters in the anticoagulation therapy group was the lowest, with no significant difference between the CDT and PCDT group.

Spectral data fusion in nondestructive detection of food products: Strategies, recent applications, and future perspectives.

In recent years, the food industry has shown a growing interest in the development of rapid and nondestructive analytical methods. However, the utilization of a solitary nondestructive detection technique offers only a constrained extent of physical or chemical insights regarding the sample under examination. To overcome this limitation, the amalgamation of spectroscopy with data fusion strategies has emerged as a promising approach. This comprehensive review delves into the fundamental principles and merits of low-level, mid-level, and high-level data fusion strategies within the domain of food analysis. Various data fusion techniques encompassing spectra-to-spectra, spectra-to-machine vision, spectra-to-electronic nose, and spectra-to-nuclear magnetic resonance are summarized. Moreover, this review also provides an overview of the latest applications of spectral data fusion techniques (SDFTs) for classification, adulteration, quality evaluation, and contaminant detection within the purview of food safety analysis. It also addresses current challenges and future prospects associated with SDFTs in real-world applications. Despite the extant technical intricacy, the ongoing evolution of online data fusion platforms and the emergence of smartphone-based multi-sensor fusion detection technology augur well for the pragmatic realization of SDFTs, endowing them with formidable capabilities for both qualitative and quantitative analysis in the realm of food analysis.

Efficient Hapten-Specific Biopanning Strategy Based on the Fe3O4@ENR-Functionalized Core-Shell Magnetic Nanoparticles.

The biopanning of target-specific phages is one of the most critical steps in the preparation of single-domain antibodies. In the traditional biopanning of haptens, the nonspecific binding of library phages to macromolecular proteins is one of the most challenging problems in preparing single-domain antibodies. In this research, Fe3O4@ENR-functionalized core-shell magnetic nanoparticles (FMNPs) were silylated and aminated by tetraethyl orthosilicate and (3-aminopropyl)triethoxysilane, and target enrofloxacin was coupled onto the surface by the carbodiimide method. The magnetic nanoparticles were characterized by Fourier transform infrared spectroscopy, particle size distribution, zeta potential, transmission electron microscopy observation, and indirect enzyme-linked immunosorbent assay (ELISA). A biopanning strategy based on Fe3O4@ENR FMNPs was then established to solve the problem in the traditional solid-phase biopanning process. The results showed that a considerable number of enrofloxacin (ENR)-positive phages were screened by only one round of biopanning. Finally, two ENR-specific shark-derived single-domain genes were identified and validated by monoclonal phage ELISA, gene sequencing, and biolayer interferometry technology. Our study provides a new biopanning strategy based on Fe3O4@ENR FMNPs for efficiently providing phages specific to haptens.

Engineered nanomaterials-based sensing systems for assessing the freshness of meat and aquatic products: A state-of-the-art review.

Meat and aquatic products are susceptible to spoilage during distribution, transportation, and storage, increasing the urgency of freshness evaluation. Engineered nanomaterials (ENMs) typically with the diameter in the range of 1-100 nm exhibit fascinating physicochemical properties. ENMs-based sensing systems have received extensive attention for food freshness assessment due to the advantages of being fast, simple, and sensitive. This review focuses on summarizing the recent application of ENMs-based sensing systems for food freshness detection. First, chemical indicators related to the freshness of meat and aquatic products are described. Then, how to apply the ENMs including noble metal nanomaterials, metal oxide nanomaterials, carbon nanomaterials, and metal-organic frameworks for the construction of different sensing systems were described. Besides, the recent advance in ENMs-based colorimetric, fluorescent, electrochemical, and surface-enhanced Raman spectroscopy sensing systems for assessing the freshness of meat and aquatic products were outlined. Finally, the challenges and future research perspectives for the application of ENMs-based sensing systems were discussed. The ENMs-based sensing systems have been demonstrated as effective tools for freshness evaluation. The sensing performance of ENMs employed in different sensing systems depends on their composition, size, shape, and stability of nanoparticles. For the real application of ENMs in food industries, the risks and regulatory issues associated with nanomaterials need to be further considered. With the continuous development of nanomaterials and sensing devices, the ENMs-based sensors are expected to be applied in-field for rapid detection of the freshness of meat and aquatic products in the future.

Light-field focusing and modulation through scattering media based on dual-polarization-encoded digital optical phase conjugation.

Digital optical phase conjugation (DOPC) can be applied for light-field focusing and imaging through or within scattering media. Traditional DOPC only recovers the phase but loses the polarization information of the original incident beam. In this Letter, we propose a dual-polarization-encoded DOPC to recover the full information (both phase and polarization) of the incident beam. The phase distributions of two orthogonal polarization components of the speckle field coming from a multimode fiber are first measured by using digital holography. Then, the phase distributions are separately modulated on two beams and their conjugations are superposed to recover the incident beam through the fiber. By changing the phase difference or amplitude ratio between the two conjugate beams, light fields with complex polarization distribution can also be generated. This method will broaden the application scope of DOPC in imaging through scattering media.

Anti-Melanogenesis and Photoprotective Effects of Ecklonia maxima Extract Containing Dieckol and Eckmaxol.

Seaweeds are potential ingredients in the cosmeceutical industry. Our previous study demonstrates that the phlorotannin-enriched extract of Ecklonia maxima (EME-EA) containing dieckol and eckmaxol possesses strong anti-inflammatory activity and suggests the cosmeceutical potential of EME-EA. In order to evaluate the cosmeceutical potential of EME-EA, the anti-melanogenesis and photoprotective effects of EME-EA were investigated in this study. EME-EA remarkably inhibited mushroom tyrosinase and melanogenesis in alpha-melanocyte-stimulating hormone-stimulated B16F10 cells. In addition, EME-EA significantly suppressed UVB-induced HaCaT cell death that was consistent with inhibition of apoptosis and reduction in scavenging intracellular reactive oxygen species. Furthermore, EME-EA significantly inhibited collagen degradation and matrix metalloproteinases expression in UVB-irradiated HDF cells in a concentration-dependent manner. These results indicate that EME-EA possesses strong anti-melanogenesis and photoprotective activities and suggest EME-EA is an ideal ingredient in the pharmaceutical and cosmeceutical industries.

Anti-Inflammatory Effect of Sulfated Polysaccharides Isolated from Codium fragile In Vitro in RAW 264.7 Macrophages and In Vivo in Zebrafish.

In this study, the anti-inflammatory activity of sulfated polysaccharides isolated from the green seaweed Codium fragile (CFCE-PS) was investigated in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages and zebrafish. The results demonstrated that CFCE-PS significantly increased the viability of LPS-induced RAW 264.7 cells in a concentration-dependent manner. CFCE-PS remarkably and concentration-dependently reduced the levels of inflammatory molecules including prostaglandin E2, nitric oxide (NO), interleukin-1 beta, tumor necrosis factor-alpha, and interleukin-6 in LPS-stimulated RAW 264.7 cells. In addition, in vivo test results indicated that CFCE-PS effectively reduced reactive oxygen species, cell death, and NO levels in LPS-stimulated zebrafish. Thus, these results indicate that CFCE-PS possesses in vitro and in vivo anti-inflammatory activities and suggest it is a potential ingredient in the functional food and pharmaceutical industries.

Identification of four hub genes in venous thromboembolism via weighted gene coexpression network analysis.

BACKGROUND: The pathogenic mechanisms of venous thromboembolism (VT) remain to be defined. This study aimed to identify differentially expressed genes (DEGs) that could serve as potential therapeutic targets for VT. METHODS: Two human datasets (GSE19151 and GSE48000) were analyzed by the robust rank aggregation method. Gene ontology and Kyoto encyclopedia of genes and genomes pathway enrichment analyses were conducted for the DEGs. To explore potential correlations between gene sets and clinical features and to identify hub genes, we utilized weighted gene coexpression network analysis (WGCNA) to build gene coexpression networks incorporating the DEGs. Then, the levels of the hub genes were analyzed in the GSE datasets. Based on the expression of the hub genes, the possible pathways were explored by gene set enrichment analysis and gene set variation analysis. Finally, the diagnostic value of the hub genes was assessed by receiver operating characteristic (ROC) analysis in the GEO database. RESULTS: In this study, we identified 54 upregulated and 10 downregulated genes that overlapped between normal and VT samples. After performing WGCNA, the magenta module was the module with the strongest negative correlation with the clinical characteristics. From the key module, FECH, GYPA, RPIA and XK were chosen for further validation. We found that these genes were upregulated in VT samples, and high expression levels were related to recurrent VT. Additionally, the four hub genes might be highly correlated with ribosomal and metabolic pathways. The ROC curves suggested a diagnostic value of the four genes for VT. CONCLUSIONS: These results indicated that FECH, GYPA, RPIA and XK could be used as promising biomarkers for the prognosis and prediction of VT.

Sparse-view imaging of a fiber internal structure in holographic diffraction tomography via a convolutional neural network.

Deep learning has recently shown great potential in computational imaging. Here, we propose a deep-learning-based reconstruction method to realize the sparse-view imaging of a fiber internal structure in holographic diffraction tomography. By taking the sparse-view sinogram as the input and the cross-section image obtained by the dense-view sinogram as the ground truth, the neural network can reconstruct the cross-section image from the sparse-view sinogram. It performs better than the corresponding filtered back-projection algorithm with a sparse-view sinogram, both in the case of simulated data and real experimental data.

Classification of cell morphology with quantitative phase microscopy and machine learning.

We describe and compare two machine learning approaches for cell classification based on label-free quantitative phase imaging with transport of intensity equation methods. In one approach, we design a multilevel integrated machine learning classifier including various individual models such as artificial neural network, extreme learning machine and generalized logistic regression. In another approach, we apply a pretrained convolutional neural network using transfer learning for the classification. As a validation, we show the performances of both approaches on classification between macrophages cultured in normal gravity and microgravity with quantitative phase imaging. The multilevel integrated classifier achieves average accuracy 93.1%, which is comparable to the average accuracy 93.5% obtained by convolutional neural network. The presented quantitative phase imaging system with two classification approaches could be helpful to biomedical scientists for easy and accurate cell analysis.

Y4-Net: a deep learning solution to one-shot dual-wavelength digital holographic reconstruction.

In this Letter, a deep learning solution (Y4-Net, four output channels network) to one-shot dual-wavelength digital holography is proposed to simultaneously reconstruct the complex amplitude information of both wavelengths from a single digital hologram with high efficiency. In the meantime, by using single-wavelength results as network ground truth to train the Y4-Net, the challenging spectral overlapping problem in common-path situations is solved with high accuracy.

Triphenylphosphine-Based Covalent Organic Frameworks and Heterogeneous Rh-P-COFs Catalysts.

The synthesis of phosphine-based functional covalent organic frameworks (COFs) has attracted great attention recently. Herein, we present two examples of triphenylphosphine-based COFs (termed P-COFs) with well-defined crystalline structures, high specific surface areas, and good thermal stability. Furthermore, rhodium catalysts with these P-COFs as support material show high turnover frequency for the hydroformylation of olefins, as well as excellent recycling performance. This work not only extends the phosphine-based COF family, but also demonstrates their application in immobilizing homogeneous metal-based (e.g., Rh-phosphine) catalysts for application in heterogeneous catalysis.

A rapid dual-channel readout approach for sensing carbendazim with 4-aminobenzenethiol-functionalized core-shell Au@Ag nanoparticles.

In this study, a 4-aminobenzenethiol-functionalized core-shell silver-coated gold nanoparticle (Au@Ag-4ABT NP) system was designed for the rapid sensing of carbendazim (CBZ) using a combination of naked-eye colorimetry and surface-enhanced Raman spectroscopy (SERS) dual-channel approach. Under alkaline conditions, the deprotonated CBZ species could interact with Ag surfaces via the N-Ag-O and N-Ag-N bonds. As a result, the neighboring Au@Ag-4ABT NPs would come closer through π-π interactions inducing the aggregation of Au@Ag-4ABT NPs. The aggregation of nanoparticles caused changes in the optical properties of the colloidal system, allowing observation with naked eyes, while the generation of more localized surface plasmon resonance "hotspots" between the adjacent Au@Ag nanoparticles permitted monitoring by the SERS technique. The proposed method showed effective sensitivity toward CBZ with limit of detection and limit of quantitation values of 37 and 122 ppb, respectively. In addition, the proposed dual-channel assay could serve as a simple and rapid method for sensing CBZ in tap water, cauliflower, and pear juice within 30 min.

Two Wnt genes regulate the expression levels of antimicrobial peptides during Vibrio infection in Macrobrachium nipponense.

The Wnt signal transduction pathway is involved in a wide variety of cellular processes, including cell proliferation, differentiation, apoptosis, and immunity against microbial infection. In the current study, we cloned and characterized two Wnt homologues (Mn-Wnt4 and Mn-Wnt16) in Macrobrachium nipponense. The full length cDNA of Mn-Wnt4 was 3144 bp with a 1074 bp open reading frame (ORF) that encoded a protein containing 358 amino acid residues. The full length cDNA of Mn-Wnt16 transcript was 2893 bp with a 1281 bp ORF that encoded a 427 amino acid protein. Mn-Wnt4 and Mn-Wnt16 proteins contained a highly conserved WNT1 domain. Tissue distribution analysis showed that Mn-Wnt4 and Mn-Wnt16 were highly expressed in the stomach. The transcriptional levels of Mn-Wnt4 and Mn-Wnt16 in the stomach were upregulated at most tested time points after bacterial (Staphylococcus aureus and Vibrio parahaemolyticus) and viral (White spot syndrome virus) infection. Moreover, the expression levels of some antimicrobial peptides (AMPs) (including anti-lipopolysaccharide factor [ALF] and crustin [CRU]) were upregulated after V. parahaemolyticus infection. We further used dsRNA-mediated RNA interference technology to explore the relationship between these two Wnt genes and the expression levels of AMPs during V. parahaemolyticus infection. Mn-Wnt4 knockdown could significantly inhibit the expression of ALF1 and CRU4 in the stomach of V. parahaemolyticus-injected prawns, whereas Mn-Wnt16 silencing could result in the inhibition of the expression level of CRU3 and CRU4 in the stomach of V. parahaemolyticus-infected prawns. These findings indicated that the Wnt gene family might participate in the body's innate immune response to Vibrio infection by regulating the synthesis of a variety of AMPs. Our study will help to understand the role of the Wnt signaling pathway in the immune response of crustaceans.

Identification of two carcinin isoforms (MnCarc1 and MnCarc2) and their function in the antimicrobial immunity of Macrobrachium nipponense.

Carcinin, a member of the crustin family, plays important roles in crustacean innate immunity. In this study, we identified two carcinin isoforms (MnCarc1 and MnCarc2) produced by alternative splicing from Macrobrachium nipponense. The full length of MnCarc1 and MnCarc2 cDNA are 1554 and 1495 bp with 687 and 609 bp open reading frame-encoding proteins that contain 228 and 202 amino acids, respectively. The genome of carcinin has nine exons and eight introns. MnCarc1 transcript contains all nine exons, whereas MnCarc2 only contains eight exons and lacks exon 4. MnCarc1 and MnCarc2 proteins contain a signal peptide, cysteine-rich regions, and a whey acidic protein domain. The phylogenetic tree shows that MnCarc1 and MnCarc2 are not grouped with other crustins and carcinins. MnCarc1 and MnCarc2 form a subgroup. MnCarc1 and MnCarc2 are widely distributed in various tissues. The expression of MnCarc1 and MnCarc2 were evidently upregulated at multiple time points in hemocytes and the intestine of M. nipponense after white spot syndrome virus, Vibrio parahaemolyticus, and Staphylococcus aureus challenges. Further studies showed that knockdown of MnDorsal or MnStat transcription factor could remarkably inhibit the upregulated expression of MnCarc1 and MnCarc2 caused by viral or bacterial challenges. In addition, recombinant MnCarc1 and MnCarc2 proteins could bind to various bacteria and polysaccharides and inhibit the growth of S. aureus and V. parahaemolyticus in vitro. This study indicated that carcinins from M. nipponense were involved in prawns innate immunity.

Activating transcription factor 2 (ATF2) negatively regulates the expression of antimicrobial peptide genes through tumor necrosis factor (TNF) in Macrobrachium nipponense.

Activating transcription factor 2 (ATF2), a member of the bZIP transcription factor family, is involved in multiple physiological and developmental processes, yet its role in the innate immunity remains unclear. In this study, two isoforms (named as MnATF2a and MnATF2b) of ATF2 gene were identified in Macrobrachium nipponense and were produced by exon skipping. The full length of MnATF2a is 2328 bp with an open reading frame of 2079 bp that encode 692 amino acids. MnATF2a has 237 bp nucleotides more than MnATF2b and the extra 237 bp is a complete exon. MnATF2a and MnATF2b proteins contain the same conserved and typical bZIP domain at the C-terminus. MnATF2a has 79 amino acids more than MnATF2b. MnATF2a and MnATF2b are widely distributed in a variety of immune tissues. After Vibrio parahaemolyticus and Staphylococcus aureus infection, the expression levels of MnATF2a and MnATF2b were significant up-regulated in the gills and stomach at 12 h. RNA interference analysis showed that knockdown of the total MnATF2 gene significantly inhibits the transcription of tumor necrosis factor (TNF) and promotes the expression of crustins (including Cru3, Cru4, and Cru7). Further study showed that knockdown of MnTNF evidently increase the expression of Cru3, Cru4, and Cru7. Our research indicates that ATF2 negatively regulate the expression of AMPs by regulating the transcription of TNF in M. nipponense. This study provides valuable information about the function of ATF2 family in the innate immunity in crustacean.