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The aim of this study was to develop a fermented food rich in β-nicotinamide mononucleotide (β-NMN). The influence of fermentation strain, fermentation time, and addition of B-group vitamins on the β-NMN content of fermented Coix (Coix lacryma-jobi) seed was investigated. Based on the results of single-factor experiments, a standardized experimental design was subsequently adopted to optimize the amount of B-group vitamins added, and to evaluate the quality and bioactivity of the fermented Coix seed. The results showed that Rhizopus oryzae was the optimal fermentation strain and the optimal fermentation time was 54 h. The optimal addition amounts of the various B-group vitamins were as follows: Nicotinamide: 150 mg/kg, niacin: 50 mg/kg, vitamin B1: 50 μg/kg. Under these conditions, the fermented Coix seed exhibited good flavor and a β-NMN content of up to 20.07 μg/g. It also contained a reducing sugar content of 19.6 g/100 g, organic acid content of 0.37 g/kg, and alcohol content of 2.5%vol. The contents of γ-aminobutyric acid, S-adenosylmethionine, and total phenols increased by 1.24, 256, and 4.7 times, respectively. The ABTS+· scavenging capacity, FRAP, ·OH scavenging capacity and DPPH· scavenging capacity increased by 14.37, 2.40, 4.55, and 4.67 times, respectively. The study provides a novel approach and valuable reference for future research on β-NMN-rich Coix functional foods.

We investigated the phenomenon of pseudo-Hopf bifurcation in a Filippov Hindmarsh-Rose neuronal system with threshold switching, and the existence of crossing limit cycles was proved by constructing the half-return mapping. Through the threshold control, the firing state of the system could be switched, allowing transitions from a non-periodic state to a periodic state, as well as the evolution from spiking to bursting. Furthermore, through threshold switching, the system exhibited the coexistence of multiple attractors, the system could be in multiple stable states, or have multiple stable sets that could attract system trajectories. This meant that neuronal system could exhibits diverse dynamical behaviors than being limited to a single stable state. The phenomenon of period-doubling bifurcation also indicated that the system will eventually enter a chaotic state. By extending the analysis to nonlinear neuronal systems, this study contributes to a deeper understanding of complex dynamics and provides valuable insights for designing state switching in the application of neural dynamics.

Abstract S100 calcium-binding protein 16 (S100A16) is implicated in both chronic kidney disease (CKD) and acute kidney injury (AKI). Previous research has shown that S100A16 contributes to AKI by facilitating the ubiquitylation and degradation of glycogen synthase kinase 3β (GSK3β) and casein kinase 1α (CK1α) through the activation of HMG-CoA reductase degradation protein 1 (HRD1). However, the mechanisms governing S100A16-induced HRD1 activation and the upregulation of S100A16 expression in renal injury are not fully understood. In this study, we observed elevated expression of Hypoxia-inducible Factor 1-alpha (HIF-1α) in the kidneys of mice subjected to ischemia-reperfusion injury (IRI). S100A16 deletion attenuated the increased HIF-1α expression induced by IRI. Using a S100A16 knockout rat renal tubular epithelial cell line (NRK-52E cells), we found that S100A16 knockout effectively mitigated apoptosis during hypoxic reoxygenation (H/R) and cell injury induced by TGF-β1. Our results revealed that H/R injuries increased both protein and mRNA levels of HIF-1α and HRD1 in renal tubular cells. S100A16 knockout reversed the expressions of HIF-1α and HRD1 under H/R conditions. Conversely, S100A16 overexpression in NRK-52E cells elevated HIF-1α and HRD1 levels. HIF-1α overexpression increased HRD1 and β-catenin while decreasing GSK-3β. HIF-1α inhibition restored HRD1 and β-catenin upregulation and GSK-3β downregulation by cellular H/R injury. Notably, Chromatin immunoprecipitation (ChIP) and luciferase reporter assays demonstrated HIF-1α binding signals on the HRD1 promoter, and luciferase reporter gene assays confirmed HIF-1α‘s transcriptional regulation of HRD1. Additionally, we identified Transcription Factor AP-2 Beta (TFAP2B) as the upregulator of S100A16. ChIP and luciferase reporter assays confirmed TFAP2B as a transcription factor for S100A16. In summary, this study identifies TFAP2B as the transcription factor for S100A16 and demonstrates HIF-1α regulation of HRD1 transcription within the S100A16-HRD1-GSK3β/CK1α pathway during renal hypoxia injury. These findings provide crucial insights into the molecular mechanisms of kidney injury, offering potential avenues for therapeutic intervention.

The idea of Pythagorean fuzzy sets (PFSs) has been extensively applied in various decision-making scenarios. Many of the applications of PFSs were carried out based on similarity functions. Some methods of similarity functions for PFSs (SFPFSs) cannot be trusted for a reliable interpretations in practical cases due to some of their setbacks. In this work, a new method of SFPFSs is developed with the capacity to outsmart the efficiency of the extant SFPFSs in terms of precise results and appropriately satisfying the rules of SFs. The new method is described with some results to validate the properties of SFs. In terms of practical application, we use the newly developed method of SFPFSs to discuss the relationship between the players of the Liverpool Football Club (FC) in the 2022/2023 English Premier League (EPL) season to assess their performances in their resurgent moments within the season. Using data from BBC Sport analysis (BBCSA) on the players' rating per match in a Pythagorean fuzzy setting, we establish the players' interactions, communications, passing, contributions, and performances to ascertain the high ranking players based on performances. Similarly, a comparative analyses are presented in tables to undoubtedly express the superiority of the newly developed method of SFPFSs. Due to the flexibility of the newly developed method of SFPFSs, it can be used for clustering analysis. In addition, the new method of SFPFSs can be extended to other uncertain environments other than PFSs.

Face recognition technology despite its extensive application across various domains. However, occlusion factors like masks and glasses are significantly impeded by current face recognition models, resulting in limitations to their practical usage. We present Occlusion-Aware Module Network called Occlusion-Aware Module-based Network (OAM-Net), designed to enhance the accuracy of occluded face recognition. OAM-Net comprises two sub-networks: an occlusion-aware sub-network and a key-region-aware sub-network. The occlusion-aware sub-network incorporates an attention module to adaptively modify the weights of convolutional kernels for optimizing the processing of occluded face images. Meanwhile, the key-region-aware sub-network integrates a Spatial Attention Residual Block (SARB) for precise identification and localization of key facial regions. The network’s generalization performance and accuracy are further enhanced by implementing a meta-learning-based strategy to boost the network’s generalization performance and accuracy. Experimental results affirm OAM-Net’s superior performance of OAM-Net over other state-of-the-art methods in occluded face recognition, underlining its significant potential for practical application.

The escalation of jellyfish stings has drawn attention to severe skin reactions, underscoring the necessity for novel treatments. This investigation assesses the potential of hydroxybenzoic acid derivatives, specifically protocatechuic acid (PCA) and gentisic acid (DHB), for alleviating Nemopilema nomurai Nematocyst Venom (NnNV)-induced injuries. By employing an in vivo mouse model, the study delves into the therapeutic efficacy of these compounds. Through a combination of ELISA and Western blot analyses, histological examinations, and molecular assays, the study scrutinizes the inflammatory response, assesses skin damage and repair mechanisms, and investigates the compounds’ ability to counteract venom effects. Our findings indicate that PCA and DHB significantly mitigate inflammation by modulating critical cytokines and pathways, altering collagen ratios through topical application, and enhancing VEGF and bFGF levels. Furthermore, both compounds demonstrate potential in neutralizing NnNV toxicity by inhibiting metalloproteinases and phospholipase-A2, showcasing the viability of small-molecule compounds in managing toxin-induced injuries.

Abstract This paper is part of a special issue on Music Technology. We study the type recognition of traditional Chinese musical instrument audio in the common way. Using MEL spectrum characteristics as input, we train an 8‐layer convolutional neural network, and finally achieve 99.3% accuracy. After that, this paper mainly studies the performance skill recognition of Chinese traditional musical instruments. Firstly, for a single instrument, the features were extracted by using the pre‐trained ResNet model, and then the SVM algorithm was used to classify all the instruments with an accuracy of 99%. Then, in order to improve the generalization of the model, the paper proposes the performance skill recognition of the same kind of instruments. In this way, the regularity of the same playing technique of different instruments can be utilized. Finally, the recognition accuracy of the four kinds of instruments is as follows: 95.7% for blowing instruments, 82.2% for plucked‐string instruments, 88.3% for strings instruments, and 97.5% for percussion instruments. We open source the audio database of traditional Chinese musical instruments and the Python source code of the whole experiment for further research.

Abstract The article presents the design and development of a virtual fretless Chinese stringed instrument App with the Duxianqin as an example, whose performance is expected to be no different from a real instrument. The digital simulation of fretless musical instruments is mainly divided into two parts: the simulation of the continuous pitch processing of the strings, and the simulation of the sound produced by plucking strings. The article returns to the theory of mechanics and wave theory and obtains the quantitative relationship between string frequency and its deformation and elongation. The Duxianqin selected in this article is a fretless instrument, which cannot be completely simulated by relying solely on sound source data. Playing and vocalization require real‐time synthesis through pitch processing, which has certain reference significance for the realization of other fretless instruments.

The incomplete silencing of exogenous transcription factors (TFs) and the lack of endogenous counterpart activation hampers the application of porcine induced pluripotent stem cells (piPSCs). We used porcine, bovine and murine TFs to reprogram porcine fetal fibroblasts. Porcine TFs-derived piPSCs (ppiPSCs) showed the highest levels of endogenous pluripotency markers activation, were able to differentiate into three germ layers and primordial germ cell-like cells (PGCLCs) and integrated into neural ectoderm of E7.5 mouse embryos in vitro. The bovine TFs derived piPSCs (bpiPSCs) expressed endogenous pluripotency markers higher than murine TFs derived piPSCs (mpiPSCs), but both had limited differentiation ability in vitro and depended on continuous expression of exogenous TFs for the maintenance. RNA sequencing confirmed ppiPSCs had distinct global transcriptional profiling, upregulated Hippo, PI3K-Akt, MAPK and relevant pluripotency signaling pathways as porcine blastocyst inner cell mass and expressed PGC early related genes. In addition, a positive and a negative correlation between exogenous and endogenous TFs’ expression level were observed in ppiPSCs and bpiPSCs lines, respectively. The TFs’ protein structures in pig were more similar to cattle than to mouse. In conclusion, the species affinity of the exogenous TFs is a key element, and the own species origin of TFs is optimal for iPSCs generation and application.

The ocean covers 71% of the surface of our planet and comprises a diverse variety of biological resources—a combination of marine animals, marine plants, and marine microorganisms that have economic value for human beings [...]

Multiple input multiple output with frequency diversity array (FDA-MIMO) radar has unique advantages in mainlobe deception jamming suppression and target location. However, if the training sample contains the target signal, it will lead to poor jamming suppression performance and large target measurement error. To deal with the problem, a method of coarse target location in the time domain is proposed based on the cumulative sampling analysis. Taking full advantages of the strongest correlation characteristic between the expected steering vector and the true target, the feature vector and feature value corresponding to the true target are found after feature decomposition. The time domain location of the target is roughly estimated during the cumulative sampling analysis from near to far. Then, a pure jamming training sample can be obtained by avoiding the location. Noise subspace projection algorithm is used to measure the angle and range of the target while suppressing mainlobe jamming. The simulation results show that the proposed method can roughly estimate the target location in the time domain when the mainlobe deception jamming completely covers the target. Compared with conventional methods, the performance of jamming suppression and target localization error are closer to the performance of ideal sampling.

Real-time continuous sensing of biological analytes is of importance in a range of biomedical applications. Here, the authors report on a flexible and physically transient sensor for the detection of nitric oxide and demonstrate applications in nitric oxide sensing in organs ex vivo and in vivo.

In this paper, based on a three-dimensional Bao system, a memristor-based hyper-chaotic Bao-like system is successfully constructed, and a simulated equivalent circuit is designed, which is used to verify the chaotic behaviors of the system. Meanwhile, a control method called periodically intermittent control with variable control width is proposed. The control width sequence in the proposed method is not only variable, but also monotonically decreasing, and the method can effectively stabilize most existing nonlinear systems. Moreover, the memristor-based hyper-chaotic Bao-like system is controlled by combining the proposed method with the Lyapunov stability principle. Finally, we should that the proposed method can effectively control and stabilize not only the proposed hyper-chaotic system, but also the Chua’s oscillator.

Jellyfish is a valuable biological resource in marine ecosystems, and blooms been observed in numerous coastal regions. However, their utility is limited by their high water content. Recent research has focused on extracting antioxidants from marine sources. In this study, we obtained jellyfish peptides (JPHT-2) through enzymatic hydrolysis of lyophilized jellyfish powder under optimal conditions and measured their antioxidant activity. Our findings indicate that JPHT-2 possesses significant radical-scavenging activity and reducing power. At a concentration of 0.74 mg/mL, JPHT-2 exhibited a remarkable ability to scavenge hydroxyl radicals, with a rate of up to 50%. The EC50 values for scavenging superoxide anion and DPPH radical were 1.55 mg/mL and 1.99 mg/mL, respectively. At the cellular level, JPHT-2 was able to protect HaCaT cells from H2O2-induced oxidative damage by increasing the level of superoxide dismutase (SOD) in cells. In conclusion, jellyfish peptides with low molecular weight can be easily obtained through hydrolysis with three enzymes and exhibit excellent antioxidant activity and safety. Jellyfish can serve as a promising source of antioxidants.

Scyphozoan Nemopilema nomurai envenomation is an unresolved threat to human health in Asian waters. Nemopilema nomurai venom metalloproteinases show important toxicities in skin damage and inflammation, but there is still no purified protein for further studies. In this study, high proteinase activity fractions in tentacle autolysis were isolated by ammonium sulfate precipitation, DEAE Sepharose Fast Flow, and Superdex 75 chromatography successively. Purification was guided by azocasein hydrolysis activity and SDS-PAGE. The final products were analyzed by LC-MS/MS. Four elution peaks purified by Superdex 75 chromatography had multiple protein bands but did not show proteinase activity. These fractions would recover proteinase activity after mixing again. Regulation mechanisms were speculated as binding metalloproteinase regulator or disaggregating metalloproteinase inhibitor by LC-MS/MS analysis. For the first time, a synergistic effect in N. nomurai proteinase activity was found in the purification process.

A developmental niche vacancy in host embryos is necessary for stem cell complementation-based organ regeneration (SCOG). Thyroid transcription factor 1 (TTF-1) is a tissue-specific transcription factor that regulates the embryonic development and differentiation of the thyroid and, more importantly, lungs; thus, it has been considered as a master gene to knockout in order to develop a lung vacancy host. TTF-1 knockout mice were originally produced by inserting a stop codon in Exon 3 of the gene (E3stop) through embryonic stem cell-based homologous recombination. The main problems of utilizing E3stop host embryos for lung SCOG are that these animals all have a tracheoesophageal fistula (TEF), which cannot be corrected by donor stem cells, and most of them have monolateral sac-like lungs. To improve the mouse model towards achieving SCOG-based lung generation, in this project, we used the CRISPR/Cas9 tool to remove Exon 2 of the gene by zygote microinjection and successfully produced TTF-1 knockout (E2del) mice. Similar to E3stop, E2del mice are birth-lethal due to retarded lung development with sac-like lungs and only a rudimentary bronchial tree, increased basal cells but without alveolar type II cells and blood vessels, and abnormal thyroid development. Unlike E3stop, 57% of the E2del embryos presented type I tracheal agenesis (TA, a kind of human congenital malformation) with a shortened trachea and clear separations of the trachea and esophagus, while the remaining 43% had TEF. Furthermore, all the E2del mice had bilateral sac-like lungs. Both TA and bilateral sac-like lungs are preferred in SCOG. Our work presents a new strategy for producing SCOG host embryos that may be useful for lung regeneration.

The development of artificial intelligence technology has made it possible to realize automatic evaluation systems for singing, and relevant research has been able to achieve accurate evaluations with respect to pitch and rhythm, but research on singing-voice timbre evaluation has remained at the level of theoretical analysis. Timbre is closely related to expression performance, breath control, emotional rendering, and other aspects of singing skills, and it has a crucial impact on the evaluation of song interpretation. The purpose of this research is to investigate the automatic evaluation method of singing-voice timbre. At the present stage, timbre research generally has problems such as a paucity of datasets, a single evaluation index, easy overfitting or a model’s failure to converge. Compared with the singing voice, the research on musical instruments is more mature, with more available data and richer evaluation dimensions. We constructed a deep network based on the CRNN model to perform timbre evaluation, and the test results showed that cross-media learning of timbre evaluation is feasible, which also indicates that humans have a consistent timbre perception with respect to musical instruments and vocals.

Nephrogenic proteins are re-expressed after ischemia/reperfusion (I/R) injury; however, the role of these proteins is still unknown. We found that sine oculis homeobox 1 (SIX1), a developmentally regulated homeoprotein, is reactivated in tubular epithelial cells after I/R injury associated with cell proliferation/migration and anti-inflammation. We demonstrated that SIX1 promoted cell proliferation by upregulating cyclin and glycolytic genes, and might increase cell migration by upregulating the expression of matrix metalloproteinase 9 (MMP9) directly or indirectly in the cell model. Notably, SIX1 targeted the promoters of the amino-terminal enhancer of split (AES) and fused in sarcoma (FUS), which are cofactors of nuclear factor-κB (NF-κB) subunit RELA, and then inhibited the transactivation function of RELA. The expression of monocyte chemotactic protein-1 (MCP-1) was decreased by the SIX1-mediated NF-κB pathway. Our results showed that the expression of cyclin, glycolytic genes, and MMP9 were significantly increased, and the infiltration of monocytes/macrophages (Mophs) was suppressed in SIX1 overexpression kidney at 1, 2, and 3 days after reperfusion. The overexpression of SIX1 resulted in reducing kidney damage from I/R injury in mice by promoting cell proliferation and migration and by inhibiting inflammation. Our study provides evidence that SIX1 involved in cell proliferation, migration, and anti-inflammation in the I/R model, which might be a potential therapeutic target that could be used to ameliorate kidney damage.

Many ion channels participate in controlling insulin synthesis and secretion of pancreatic β-cells. Epithelial sodium channel (ENaC) expressed in human pancreatic tissue, but the biological role of ENaC in pancreatic β-cells is still unclear. Here, we applied the CRISPR/Cas9 gene editing technique to knockout α-ENaC gene in a murine pancreatic β-cell line (MIN6 cell). Four single-guide RNA (sgRNA) sites were designed for the exons of α-ENaC. The sgRNA1 and sgRNA3 with the higher activity were constructed and co-transfected into MIN6 cells. Through processing a series of experiment flow included drug screening, cloning, and sequencing, the α-ENaC gene-knockout (α-ENaC−/−) in MIN6 cells were obtained. Compared with the wild-type MIN6 cells, the cell viability and insulin content were significantly increased in α-ENaC−/− MIN6 cells. Therefore, α-ENaC−/− MIN6 cells generated by CRISPR/Cas9 technology added an effective tool to study the biological function of α-ENaC in pancreatic β-cells.

Transient electronics (or biodegradable electronics) is an emerging technology whose key characteristic is an ability to dissolve, resorb, or physically disappear in physiological environments in a controlled manner. Potential applications include eco-friendly sensors, temporary biomedical implants, and data-secure hardware. Biodegradable electronics built with water-soluble, biocompatible active and passive materials can provide multifunctional operations for diagnostic and therapeutic purposes, such as monitoring intracranial pressure, identifying neural networks, assisting wound healing process, etc. This review summarizes the up-to-date materials strategies, manufacturing schemes, and device layouts for biodegradable electronics, and the outlook is discussed at the end. It is expected that the translation of these materials and technologies into clinical settings could potentially provide vital tools that are beneficial for human healthcare. Keywords: Biodegradable electronics, Transient electronics, Biodegradable materials, Silicon, Metals

Abstract Complement protein C3 is the pivotal component of the complement system. Previous studies have demonstrated that C3 has implications in various human diseases and exerts profound functions under certain conditions. However, the delineation of pathological and physiological roles of C3 has been hampered by the insufficiency of suitable animal models. In the present study, we applied the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system to target the C3 gene in porcine fetal fibroblasts. Our results indicated that CRISPR/Cas9 targeting efficiency was as high as 84.7%, and the biallelic mutation efficiency reached at 45.7%. The biallelic modified colonies were used as donor for somatic cell nuclear transfer (SCNT) technology to generate C3 targeted piglets. A total of 19 C3 knockout (KO) piglets were produced and their plasma C3 protein was undetectable by western blot analysis and ELISA. The hemolytic complement activity and complement-dependent cytotoxicity assay further confirmed that C3 was disrupted in these piglets. These C3 KO pigs could be utilized as a valuable large animal model for the elucidation of the roles of C3.

The suppression of mainlobe jamming (MLJ) is a hard task and an open problem in the electronic counter-countermeasures field. In this article, a sparse Bayesian learning (SBL)-based mainlobe blanket jamming suppression algorithm is proposed for solving this problem, which can effectively suppress the MLJ and estimate the target distance and direction. The proposed algorithm can be divided into two steps. First, the row (or column) spatial adaptive processing is adopted to suppress the MLJ; Second, the SBL algorithm is used to obtain the target echo parameters.

Jellyfish are rich in resources and widely distributed along coastal areas. As a potential approach to respond to jellyfish blooms, the use of jellyfish-derived products is increasing. The citrus spider mite (Panonychus citri) is one of the key citrus pests, negatively impacting the quality and quantity of oranges. Due to the resistance and residue of chemical acaricides, it is important to seek natural substitutes that are environmentally friendly. The field efficacy of the venom from the jellyfish Nemopilema nomurai against P. citri was assayed in a citrus garden. The frozen N. nomurai tentacles were sonicated in different buffers to isolate the venom. The venom isolated by PBS buffer (10 mM, pH 6.0) had the strongest acaricidal activity of the four samples, and the corrected field efficacy 7 days after treatment was up to 95.21%. This study demonstrated that jellyfish has potential use in agriculture.

As a popular marine saccharide, chitooligosaccharides (COS) has been proven to have good antioxidant activity. Its antioxidant effect is closely related to its degree of polymerization, degree of acetylation and sequence. However, the specific structure–activity relationship remains unclear. In this study, three chitosan dimers with different sequences were obtained by the separation and enzymatic method, and the antioxidant activity of all four chitosan dimers were studied. The effect of COS sequence on its antioxidant activity was revealed for the first time. The amino group at the reducing end plays a vital role in scavenging superoxide radicals and in the reducing power of the chitosan dimer. At the same time, we found that the fully deacetylated chitosan dimer DD showed the strongest DPPH scavenging activity. When the amino groups of the chitosan dimer were acetylated, it showed better activity in scavenging hydroxyl radicals. Research on COS sequences opens up a new path for the study of COS, and is more conducive to the investigation of its mechanism.

Jellyfish stings are a common issue globally, particularly in coastal areas in the summer. Victims can suffer pain, itching, swelling, shock, and even death. Usually, hot water, vinegar, or alumen is used to treat the normal symptoms of a jellyfish sting. However, a specific antivenom may be an effective treatment to deal with severe jellyfish stings. Cyanea nozakii often reach a diameter of 60 cm and are responsible for hundreds of thousands of stings per year in coastal Chinese waters. However, there has been no specific C. nozakii antivenom until now, and so the development of this antivenom is very important. Herein, we collected C. nozakii antisera from tentacle extract venom immunized rabbits and purified the immunoglobulin (IgG) fraction antivenom (AntiCnTXs). Subsequently, two complete procedures to produce a refined F(ab’)2 type of antivenom (F(ab’)2-AntiCnTXs) and Fab type of antivenom (Fab-AntiCnTXs) by multiple optimizations and purification were established. The neutralization efficacy of these three types of antivenoms was compared and analyzed in vitro and in vivo, and the results showed that all types of antibodies displayed some neutralization effect on the lethality of C. nozakii venom toxins, with the neutralization efficacy as follows: F(ab’)2-AntiCnTXs ≥ AntiCnTXs > Fab-AntiCnTXs. This study describes the preparation of novel C. nozakii jellyfish antivenom preparations towards the goal of developing a new, effective treatment for jellyfish stings.

Scyphozoan envenomation is featured as severe cutaneous damages due to the toxic effects of venom components released by the stinging nematocysts of a scyphozoan. However, the oedematogenic property and mechanism of scyphozoan venoms remain uninvestigated. Here, we present the oedematogenic properties of the nematocyst venom from Nemopilema nomurai (NnNV), a giant stinging scyphozoan in China, for the first time, using in vivo and in vitro models with class-specific inhibitors. NnNV was able to induce remarkable oedematogenic effects, including induction of significant oedema in the footpad and thigh of mouse, and increase in vascular permeability in the dorsal skin and kidney. Moreover, batimastat, a specific metalloproteinase inhibitor, could significantly reduce the Evan’s blue leakage in the damaged organs and attenuate paw oedema after 12 h, but exerted no influence on NnNV-induced thigh oedema. These observations suggested a considerable contribution of NnNV metalloproteinase-like components to the increased vasopermeability, and the participation was strongly suggested to be mediated by destroying the integrity of the vascular basement membrane. Moreover, partial isolation combined LC-MS/MS profiling led to identification of the protein species Nn65 with remarkable metalloproteinase activity. This study contributes to the understanding of the effector components underlying the cutaneous damages induced by scyphozoan stings.

Miniature pigs have advantages over rodents in modeling atherosclerosis because their cardiovascular system and physiology are similar to that of humans. Apolipoprotein E (ApoE) deficiency has long been implicated in cardiovascular disease in humans. To establish an improved large animal model of familial hypercholesterolemia and atherosclerosis, the clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 system (CRISPR/Cas9) was used to disrupt the ApoE gene in Bama miniature pigs. Biallelic-modified ApoE pigs with in-frame mutations (ApoEm/m) and frameshift mutations (ApoE−/−) were simultaneously produced. ApoE−/− pigs exhibited moderately increased plasma cholesterol levels when fed with a regular chow diet, but displayed severe hypercholesterolemia and spontaneously developed human-like atherosclerotic lesions in the aorta and coronary arteries after feeding on a high-fat and high-cholesterol (HFHC) diet for 6 months. Thus, these ApoE−/− pigs could be valuable large animal models for providing further insight into translational studies of atherosclerosis.

The silkworm Bombyx mori L. (B. mori) has a significant impact on the economy by producing more than 80% of the globally produced raw silk. The exposure of silkworm to pesticides may cause adverse effects on B. mori, such as a reduction in the production and quality of silk. This study aims to assay the effect of venom from the jellyfish Nemopilema nomurai on growth, cuticle and acetylcholinesterase (AChE) activity of the silkworm B. mori by the leaf dipping method. The experimental results revealed that the four samples caused neither antifeeding nor a lethal effect on B. mori. The sample SFV inhibited B. mori growth after 6 days of treatment in a dose-dependent manner. The samples SFV, DSFV and Fr-1 inhibited the precipitation and synthesis of chitin in the cuticle after 12 and 14 days of treatment. In the case of the four samples, the AChE was significantly improved after 14 days of treatment.

The present study investigated and compared the hypoglycemic activity of differently regioselective chitosan sulfates in alloxan-induced diabetic rats. Compared with the normal control rats, significantly higher blood glucose levels were observed in the alloxan-induced diabetic rats. The differently regioselective chitosan sulfates exhibited hypoglycemic activities at different doses and intervals, especially 3-O-sulfochitosan (3-S). The major results are as follows. First, 3,6-di-O-sulfochitosan and 3-O-sulfochitosan exhibited more significant hypoglycemic activities than 2-N-3, 6-di-O-sulfochitosan and 6-O-sulfochitosan. Moreover, 3-S-treated rats showed a more significant reduction of blood glucose levels than those treated by 3,6-di-O-sulfochitosan. These results indicated that –OSO3− at the C3-position of chitosan is a key active site. Second, 3-S significantly reduced the blood glucose levels and regulated the glucose tolerance effect in the experimental rats. Third, treatment with 3-S significantly increased the plasma insulin levels in the experimental diabetic rats. A noticeable hypoglycemic activity of 3-S in the alloxan-induced diabetic rats was shown. Clinical trials are required in the future to confirm the utility of 3-S.

Jellyfish envenomations result in extensive dermatological symptoms, clinically named as jellyfish dermatitis, which can seriously affect the daily activities and physical health of people. Inflammatory response accompanies the whole process of jellyfish dermatitis and the complexity of jellyfish venom components makes it difficult to treat jellyfish dermatitis symptoms effectively. Moreover, inhibiting inflammation is essential for the treatment of jellyfish stings and exploring the main components of jellyfish venom that cause inflammation is an urgent research area. In this study, the inhibitory effects of matrix metalloproteinase (MMP) inhibitors for venom-induced inflammation were explored at a cellular level. The expression of the three inflammatory factors, IL-6, TNF-α and MCP-1 in two skin cell lines, human keratinocyte cells (HaCaT) and human embryonic skin fibroblasts cells (CCC-ESF-1), at the cellular level, after treatment with the inhibitors of jellyfish Nemopilema nomurai (N. nomurai) nematocyst venom (NnNV-I), were determined. The results showed that inhibitors of MMP can significantly reduce the toxic effects of jellyfish Nemopilema nomurai nematocyst venom (NnNV) to skin cells. The expression levels of the three inflammatory factors IL-6, MCP-1, and TNF-α in the cells were also significantly decreased, indicating that MMPs in jellyfish venom are probably vital factors leading to jellyfish dermatitis. This study is beneficial in the prevention and treatment of jellyfish stings.

Recently, Tribonema sp., a kind of filamentous microalgae, has been studied for biofuel production due to its accumulation of triacylglycerols. However, the polysaccharides of Tribonema sp. and their biological activities have rarely been reported. In this paper, we extracted sulfated polysaccharides from Tribonema sp. (TSP), characterized their chemical composition and structure, and determined their immunostimulation and anticancer activities on RAW264.7 macrophage cells and HepG2 cells. The results showed that TSP is a sulfated polysaccharide with a Mw of 197 kDa. TSP is a heteropolysaccharide that is composed mainly of galactose. It showed significant immune-modulatory activity by stimulating macrophage cells, such as upregulating interleukin 6 (IL-6), interleukin 10 (IL-10), and tumor necrosis factor α (TNF-α). In addition, TSP also showed significant dose-dependent anticancer activity (with an inhibition rate of up to 66.8% at 250 µg/mL) on HepG2 cells as determined by the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The cycle analysis indicated that the anticancer activity of TSP is mainly the result of induced cell apoptosis rather than affecting the cell cycle and mitosis of HepG2 cells. These findings suggest that TSP might have potential as an anticancer resource, but further research is needed, especially in vivo experiments, to explore the anticancer mechanism of TSP.

Trophoblasts (TR) are specialized cells of the placenta and play an important role in embryo implantation. The in vitro culture of trophoblasts provided an important tool to investigate the mechanisms of implantation. In the present study, porcine trophoblast cells were derived from pig in vitro fertilized (IVF) and parthenogenetically activated (PA) blastocysts via culturing in medium supplemented with KnockOut serum replacement (KOSR) and basic fibroblast growth factor (bFGF) on STO feeder layers, and the effect of ROCK (Rho-associated coiled-coil protein kinases) inhibiter Y-27632 on the cell lines culture was tested. 5 PA blastocyst derived cell lines and 2 IVF blastocyst derived cell lines have been cultured more than 20 passages; one PA cell lines reached 110 passages without obvious morphological alteration. The derived trophoblast cells exhibited epithelium-like morphology, rich in lipid droplets, and had obvious defined boundaries with the feeder cells. The cells were histochemically stained positive for alkaline phosphatase. The expression of TR lineage markers, such as CDX2, KRT7, KRT18, TEAD4, ELF5 and HAND1, imprinted genes such as IGF2, PEG1 and PEG10, and telomerase activity related genes TERC and TERF2 were detected by immunofluorescence staining, reverse transcription PCR and quantitative real-time PCR analyses. Both PA and IVF blastocysts derived trophoblast cells possessed the ability to differentiate into mature trophoblast cells in vitro. The addition of Y-27632 improved the growth of both PA and IVF blastocyst derived cell lines and increased the expression of trophoblast genes. This study has provided an alternative highly efficient method to establish trophoblast for research focused on peri-implantation and placenta development in IVF and PA embryos.

Fungi isolated from marine invertebrates are of considerable importance as new promising sources of unique secondary metabolites with significant biomedical potential. However, the cultivable fungal community harbored in jellyfish was less investigated. In this work, we seek to recover symbiotic fungi from different tissues of jellyfish Nemopilema nomurai. A total of seven morphotypes were isolated, which were assigned into four genera (Aspergillus, Cladosporium, Purpureocillium, and Tilletiopsis) from two phyla (Ascomycota and Basidiomycota) by comparing the rDNA-ITS sequences with the reference sequences in GenBank. The most fungi were found in the inner tissues of subumbrella. Two of the cultivation-independent procedures, changing media type and co-cultivation, were employed to maximize the complexity of metabolites. Thus, thirteen EtOAc gum were obtained and fingerprinted by High Performance Liquid Chromatography (HPLC) equipped with a photodiode array (PDA) detector. Antibacterial and antifungal activities of these complex mixtures were tested against a panel of bacterial and fungal pathogens. The antimicrobial results showed that all of the 13 EtOAc extracts displayed different levels of antibacterial activity, three of which exhibited strong to significant antibacterial activity to the bacterial pathogens Staphylococcus aureus and Salmonella entrica. Antifungal activity indicated that the EtOAc extracts from pure culture of Aspergillus versicolor and co-culture of A. versicolor and Tilletiopsis sp. in rice media were promising for searching new compounds, with the maximal mycelial growth inhibition of 82.32% ± 0.61% for Rhizoctonia solani and 48.41% ± 11.02% for Botrytis cinerea at 200 μg/ml, respectively. This study is the first report on the antibacterial and antifungal activity of jellyfish-associated fungi and allows the first sight into cultivable fungal community residing in jellyfish. Induced metabolites by cultivation-dependent approaches provides a new reservoir for drug discovery from jellyfish-derived fungi.

Mammalian haploid cell lines provide useful tools for both genetic studies and transgenic animal production. To derive porcine haploid cells, three sets of experiments were conducted. First, genomes of blastomeres from 8-cell to 16-cell porcine parthenogenetically activated (PA) embryos were examined by chromosome spread analysis. An intact haploid genome was maintained by 48.15% of blastomeres. Based on this result, two major approaches for amplifying the haploid cell population were tested. First, embryonic stem-like (ES-like) cells were cultured from PA blastocyst stage embryos, and second, fetal fibroblasts from implanted day 30 PA fetuses were cultured. A total of six ES-like cell lines were derived from PA blastocysts. No chromosome spread with exactly 19 chromosomes (the normal haploid complement) was found. Four cell lines showed a tendency to develop to polyploidy (more than 38 chromosomes). The karyotypes of the fetal fibroblasts showed different abnormalities. Cells with 19-38 chromosomes were the predominant karyotype (59.48-60.91%). The diploid cells were the second most observed karyotype (16.17%-22.73%). Although a low percentage (3.45-8.33%) of cells with 19 chromosomes were detected in 18.52% of the fetus-derived cell lines, these cells were not authentic haploid cells since they exhibited random losses or gains of some chromosomes. The haploid fibroblasts were not efficiently enriched via flow cytometry sorting. On the contrary, the diploid cells were efficiently enriched. The enriched parthenogenetic diploid cells showed normal karyotypes and expressed paternally imprinted genes at extremely low levels. We concluded that only a limited number of authentic haploid cells could be obtained from porcine cleavage-stage parthenogenetic embryos. Unlike mouse, the karyotype of porcine PA embryo-derived haploid cells is not stable, long-term culture of parthenogenetic embryos, either in vivo or in vitro, resulted in abnormal karyotypes. The porcine PA embryo-derived diploid fibroblasts enriched from sorting might be candidate cells for paternally imprinted gene research.

Background: Medusozoans utilize explosively discharging penetrant nematocysts to inject venom into prey. These venoms are composed of highly complex proteins and peptides with extensive bioactivities, as observed in vitro. Diverse enzymatic toxins have been putatively identified in the venom of jellyfish, Nemopilema nomurai and Cyanea nozakii, through examination of their proteomes and transcriptomes. However, functional examination of putative enzymatic components identified in proteomic approaches to elucidate potential bioactivities is critically needed. Methods: In this study, enzymatic toxins were functionally identified using a combined approach consisting of in gel zymography and liquid chromatography tandem mass spectrometry (LC-MS/MS). The potential roles of metalloproteinases and lipases in hemolytic activity were explored using specific inhibitors. Results: Zymography indicated that nematocyst venom possessed protease-, lipase- and hyaluronidase-class activities. Further, proteomic approaches using LC-MS/MS indicated sequence homology of proteolytic bands observed in zymography to extant zinc metalloproteinase-disintegrins and astacin metalloproteinases. Moreover, pre-incubation of the metalloproteinase inhibitor batimastat with N. nomurai nematocyst venom resulted in an approximate 62% reduction of hemolysis compared to venom exposed sheep erythrocytes, suggesting that metalloproteinases contribute to hemolytic activity. Additionally, species within the molecular mass range of 14–18 kDa exhibited both egg yolk and erythrocyte lytic activities in gel overlay assays. Conclusion: For the first time, our findings demonstrate the contribution of jellyfish venom metalloproteinase and suggest the involvement of lipase species to hemolytic activity. Investigations of this relationship will facilitate a better understanding of the constituents and toxicity of jellyfish venom.

The carmine spider mite Tetranychus cinnabarinus (T. cinnabarinus) is a common polyphagous pest that attacks crops, vegetables, flowers, and so on. It is necessary to find lead compounds for developing novel, powerful, and environmentally-friendly acaricides as an alternative approach to controlling the carmine spider mite because of the serious resistance and residual agrochemicals in the environment. In addition, the study on the acaricidal activities of marine bioactive substances is comparatively deficient. In the present study, the acaricidal activity of venom (NnFV) from the jellyfish Nemopilema nomurai against the carmine spider mite T. cinnabarinus was determined for the first time. The venom had contact toxicity, and the 24-h LC50-value was 29.1 μg/mL. The mite body wall was affected by the venom, with the mite body having no luster and being seriously shrunken after 24 h. T. cinnabarinus was a potential target pest of NnFV, which had potential as a type of natural bioacaricide. The repellent activity and systemic toxicity of the venom against T. cinnabarinus were also studied. However, NnFV had no repellent activity and systemic toxicity against T. cinnabarinus.