Sulfonamide modified chitosan oligosaccharide with high nematicidal activity against Meloidogyne incognita.

Chitosan oligosaccharide (COS) modification is a feasible way to develop novel green nematicides. This study involved the synthesis of various COS sulfonamide derivatives via hydroxylated protection and deprotection, which were then characterized using NMR, FTIR, MS, elemental analysis, XRD, and TG/DTG. In vitro experiments found that COS-alkyl sulfonamide derivatives (S6 and S11-S13) exhibited high mortality (>98 % at 1 mg/mL) against Meloidogyne incognita second-instar larvaes (J2s) among the derivatives. S6 can cause vacuole-like structures in the middle and tail regions of the nematode body and effectively inhibit egg hatching. In vivo tests have found that S6 has well control effects and low plant toxicity. Additionally, the structure-activity studies revealed that S6 with a high degree of substitution, a low molecular weight, and a sulfonyl bond on the amino group of the COS backbone exhibited increased nematicidal activity. The sulfonamide group is a potential active group for developing COS-based nematicides.

Intermediate molecular weight-fucosylated chondroitin sulfate from sea cucumber Cucumaria frondosa is a promising anticoagulant targeting intrinsic factor IXa.

Thromboembolic diseases pose a serious risk to human health worldwide. Fucosylated chondroitin sulfate (FCS) is reported to have good anticoagulant activity with a low bleeding risk. Molecular weight plays a significant role in the anticoagulant activity of FCS, and FCS smaller than octasaccharide in size has no anticoagulant activity. Therefore, identifying the best candidate for developing novel anticoagulant FCS drugs is crucial. Herein, native FCS was isolated from sea cucumber Cucumaria frondosa (FCScf) and depolymerized into a series of lower molecular weights (FCScfs). A comprehensive assessment of the in vitro anticoagulant activity and in vivo bleeding risk of FCScfs with different molecule weights demonstrated that 10 kDa FCScf (FCScf-10 K) had a greater intrinsic anticoagulant activity than low molecular weight heparin (LMWH) without any bleeding risk. Using molecular modeling combined with experimental validation, we revealed that FCScf-10 K can specifically inhibit the formation of the Xase complex by binding the negatively charged sulfate group of FCScf-10 K to the positively charged side chain of arginine residues on the specific surface of factor IXa. Thus, these data demonstrate that the intermediate molecular weight FCScf-10 K is a promising candidate for the development of novel anticoagulant drugs.

In vivo immunological activity of chitosan-derived nanoparticles.

Chitosan has been studied as an immunomodulator, but few studies have used chitosan derivatives as adjuvants alone. After a preliminary study, we found that nanoparticles prepared from chitosan derivatives had better cellular immune activity when used as an adjuvant. Therefore, animal experiments were conducted to further investigate the performance and mechanism of these nanoparticles as immune adjuvants. We injected mice with the chitosan nanoparticle vaccine and measured the expression levels of immunoglobulins, immune factors, and immune genes in tissues and tissue sections. The results showed that C236-HACC-OVA (C2,3,6-chitosan sulfate-chitosan quaternary ammonium salt-ovalbumin) and NO-HACC-OVA (NO-carboxymethyl chitosan-chitosan quaternary ammonium salt-ovalbumin) nanoparticles can significantly improve the secretion of the immune factors IL-6, TNF, and IL-1ß. The level of IgG1 was highly significant after administering both nanoparticles, but IgG2 was not significant in mice. Three immune factors (IL-4, IL-6, and IL-17) were secreted at high levels in mouse serum at a nanoparticle dose of 0.3 mg/mouse. These nanoparticles also have high safety in the liver, kidney, and spleen of mice. This study proves the possibility of using chitosan derivative nanoparticles as vaccine adjuvants. These data further indicate that chitosan derivative nanoparticles have potential for use as vaccine adjuvants and demonstrate that polysaccharides have a unique position in green vaccine research.

Synthesis and Characterization of Slow-Release Chitosan Oligosaccharide Pyridine Schiff Base Copper Complexes with Antifungal Activity.

Herein, a series of chitosan oligosaccharide copper complexes modified with pyridine groups (CPSx-Cu complexes) were successfully prepared via the Schiff base reaction and ion complexation reaction for slow-release fungicide. The structures of the synthesized derivatives were characterized via Fourier transform infrared spectroscopy and 1H and 13C nuclear magnetic resonance spectroscopy, and the unit configuration of the complexes was calculated using Gaussian software. The slow-release performance experiment demonstrated that the cumulative copper ion release rate of CPSx-Cu complexes was dependent on the type of substituents on the pyridine ring. Furthermore, the in vitro and in vivo antifungal activities of the CPSx-Cu complexes were investigated. At a concentration of 0.4 mg/mL, CPSx-Cu complexes completely inhibited the growth of Pythium vexans and Phytophthora capsici. Results indicated that CPSx-Cu complexes with slow-release ability exhibited better antifungal activity than thiodiazole-copper and copper sulfate basic. This study confirmed that combining chitosan oligosaccharide with bioactive pyridine groups and copper ions is an effective approach to further developing slow-release copper fungicides, providing new possibilities for the application of copper fungicides in green agriculture. This study lays the foundation for further studies on biogreen copper fungicides.

Effects of toxin metalloproteinases from jellyfish Nemopilema nomurai nematocyst on the dermal toxicity and potential treatment of jellyfish dermatitis.

Jellyfish dermatitis is a common medical problem in many countries due to the jellyfish envenomation. However, there are no specific and targeted medications for their treatment. Here we investigated the possible therapeutic effects of metalloproteinase inhibitors on the dermal toxicity of Nemopilema nomurai nematocyst venom (NnNV), a giant venomous jellyfish from China, using the jellyfish dermatitis model, focusing on inflammatory effector molecules during jellyfish envenomation. Metalloproteinase may further stimulate inflammation by promoting oxidative stress in the organism and play key roles by activating MAPK and NF-κB, in the pathogenesis of jellyfish dermatitis. And the metalloproteinase inhibitors batimastat and EDTA disodium salt may treat the Jellyfish dermatitis by inhibiting the metalloproteinase activity in NnNV. These observations suggest that the metalloproteinase components of NnNV make a considerable contribution to dermal toxicity as the inflammation effect molecular, and metalloproteinase inhibitors can be regarded as novel therapeutic medicines in jellyfish envenomation. This study contributes to understanding the mechanism of jellyfish dermatitis and suggests new targets and ideas for the treatment of jellyfish envenomation.

Toxin metalloproteinases exert a dominant influence on pro-inflammatory response and anti-inflammatory regulation in jellyfish sting dermatitis.

Toxin metalloproteinases are the primary components responsible for various toxicities in jellyfish venom, and there is still no effective specific therapy for jellyfish stings. The comprehension of the pathogenic mechanisms underlying toxin metalloproteinases necessitates further refinement. In this study, we conducted a differential analysis of a dermatitis mouse model induced by jellyfish Nemopilema nomurai venom (NnNV) samples with varying levels of metalloproteinase activity. Through skin tissue proteomics and serum metabolomics, the predominant influence of toxin metalloproteinase activity on inflammatory response was revealed, and the signal pathway involved in its regulation was identified. In skin tissues, many membrane proteins were significantly down-regulated, which might cause tissue damage. The expression of pro-inflammatory factors was mainly regulated by PI3K-Akt signaling pathway. In serum, many fatty acid metabolites were significantly down-regulated, which might be the anti-inflammation feedback regulated by NF-κB p65 signaling pathway. These results reveal the dermatitis mechanism of toxin metalloproteinases and provide new therapeutic targets for further studies. SIGNIFICANCE: Omics is an important method to analyze the pathological mechanism and discover the key markers, which can reveal the pathological characteristics of jellyfish stings. Our research first analyzed the impact of toxin metalloproteinases on jellyfish sting dermatitis by skin proteomics and serum metabolomics. The present results suggest that inhibition of toxin metalloproteinases may be an effective treatment strategy, and provide new references for further jellyfish sting studies.

Therapeutic potential of enzymatically extracted eumelanin from squid ink in type 2 diabetes mellitus ICR mice: Multifaceted intervention against hyperglycemia, oxidative stress and depression.

BACKGROUND: Type 2 diabetes mellitus (T2DM) is a prevalent metabolic disease that poses significant health risks due to its numerous complications. However, the effects of eumelanin on oxidative stress, hyperglycemia and depression in diabetic mice have not been extensively studied. RESULTS: Our study employed an enzymatic approach to extract eumelanin from squid ink and characterized it using spectroscopic techniques. Remarkably, eumelanin extracted with alkaline-neutral-flavor protease (ANF) displayed superior inhibitory activity against α-glucosidase and α-amylase, while enhancing glucose utilization and hepatic glycogen synthesis in human hepatocellular carcinoma cell line (HepG2) insulin resistance model. Further evaluation of ANF in a T2DM ICR mouse model demonstrated its significant potential in alleviating hyperglycemia, reducing glycosylated serum protein levels, improving glucose tolerance and modulating total cholesterol and low-density lipoprotein levels, as well as antioxidant indices at a dosage of 0.04 g kg-1 . Additionally, ANF exhibited positive effects on energy levels and reduced immobility time in antidepressant behavioral experiments. Moreover, ANF positively influenced the density and infiltration state of renal cells, while mitigating inflammatory enlargement and deformation of liver cells, without inducing any adverse effects in mice. CONCLUSION: Overall, these findings underscore the significant therapeutic potential of ANF in the treatment of T2DM and its associated complications. By augmenting lipid and glucose metabolism, mitigating oxidative stress and alleviating depression, ANF emerges as a promising candidate for multifaceted intervention. © 2023 Society of Chemical Industry.

Fucoidan may treat jellyfish dermatitis by inhibiting the inflammatory effect of jellyfish venom.

Jellyfish dermatitis is a common medical problem caused by jellyfish stings. However, there are no targeted and effective medications for their treatment. Here, the biological activity of fucoidan for treatment of jellyfish dermatitis was investigated for the first time. 3 mg/mL Fucoidan attenuated the inflammatory effects of Nemopilema nomurai nematocyst venom (NnNV), including dermal toxicity and myotoxicity. Fucoidan may decrease the inflammatory effects of NnNV by downregulating MAPK and NF-κB pathways. This may be attributed to the inhibitory effect of fucoidan on metalloproteinases and phospholipase A2 (PLA2) in NnNV. 3 mg/mL fucoidan reduced the metalloproteinase activity in NnNV from 316.33 ± 20.84 U/mg to 177.33 ± 25.36 U/mg, while the inhibition of PLA2 activity in NnNV by 1 mg/mL fucoidan could reach 37.67 ± 3.42 %. Besides, external application of 3 mg/mL fucoidan can effectively alleviate the symptoms of jellyfish dermatitis. These observations suggest that fucoidan has considerable potential for treatment of jellyfish dermatitis and could be regarded as a novel medicine for jellyfish envenomation. This study provides new ideas for treatment of jellyfish envenomation and suggests evidence for the use of fucoidan in the treatment of jellyfish dermatitis as well as broadens the potential application of fucoidan in clinical practice.

The source, activity influencing factors and biological activities for future development of chitin deacetylase.

Chitin deacetylase (CDA), a prominent member of the carbohydrate esterase enzyme family 4 (CE4), is found ubiquitously in bacteria, fungi, insects, and crustaceans. This metalloenzyme plays a pivotal role in recognizing and selectively removing acetyl groups from chitin, thus offering an environmentally friendly and biologically-driven preparation method for chitosan with immense industrial potential. Due to its diverse origins, CDAs sourced from different organisms exhibit unique functions, optimal pH ranges, and temperature preferences. Furthermore, certain organic reagents can induce structural changes in CDAs, influencing their catalytic activity. Leveraging CDA's capabilities extends beyond chitosan biocatalysis, as it demonstrates promising application value in agricultural pest control. In this paper, the source, reaction mechanism, influencing factors, the fermentation methods and applications of CDA are reviewed, which provides theoretical help for the research and application of CDA.

Comparative analysis of PacBio and ONT RNA sequencing methods for Nemopilema Nomurai venom identification.

Recent studies on marine organisms have made use of third-generation sequencing technologies such as Pacific Biosciences (PacBio) and Oxford Nanopore Technologies (ONT). While these specialized bioinformatics tools have different algorithmic designs and performance capabilities, they offer scalability and can be applied to various datasets. We investigated the effectiveness of PacBio and ONT RNA sequencing methods in identifying the venom of the jellyfish species Nemopilema nomurai. We conducted a detailed analysis of the sequencing data from both methods, focusing on key characteristics such as CD, alternative splicing, long-chain noncoding RNA, simple sequence repeat, transcription factor, and functional transcript annotation. Our findings indicate that ONT generally produced higher raw data quality in the transcriptome analysis, while PacBio generated longer read lengths. PacBio was found to be superior in identifying CDs and long-chain noncoding RNA, whereas ONT was more cost-effective for predicting alternative splicing events, simple sequence repeats, and transcription factors. Based on these results, we conclude that PacBio is the most specific and sensitive method for identifying venom components, while ONT is the most cost-effective method for studying venogenesis, cnidocyst (venom gland) development, and transcription of virulence genes in jellyfish. Our study has implications for future sequencing technologies in marine jellyfish, and highlights the power of full-length transcriptome analysis in discovering potential therapeutic targets for jellyfish dermatitis.

Melanin: insights into structure, analysis, and biological activities for future development.

Melanin, a widely distributed pigment found in various organisms, possesses distinct structures that can be classified into five main types: eumelanin (found in animals and plants), pheomelanin (found in animals and plants), allomelanin (found in plants), neuromelanin (found in animals), and pyomelanin (found in fungi and bacteria). In this review, we present an overview of the structure and composition of melanin, as well as the various spectroscopic identification methods that can be used, such as Fourier transform infrared (FTIR) spectroscopy, electron spin resonance (ESR) spectroscopy, and thermogravimetric analysis (TGA). We also provide a summary of the extraction methods of melanin and its diverse biological activities, including antibacterial properties, anti-radiation effects, and photothermal effects. The current state of research on natural melanin and its potential for further development is discussed. In particular, the review provides a comprehensive summary of the analysis methods used to determine melanin species, offering valuable insights and references for future research. Overall, this review aims to provide a thorough understanding of the concept and classification of melanin, its structure, physicochemical properties, and structural identification methods, as well as its various applications in the field of biology.

A Computational Biology Study on the Structure and Dynamics Determinants of Thermal Stability of the Chitosanase from Aspergillus fumigatus.

Environmentally friendly and efficient biodegradation with chitosanase for degrading chitosan to oligosaccharide has been gaining more importance. Here, we studied a chitosanase from Aspergillus fumigatus with potential for production, but does not have the ideal thermal stability. The structure predicted by the Alphafold2 model, especially the binding site and two catalytic residues, has been found to have a high similarity with the experimental structure of the chitosanase V-CSN from the same family. The effects of temperature on structure and function were studied by dynamic simulation and the results showed that the binding site had high flexibility. After heating up from 300 K to 350 K, the RMSD and RMSF of the binding site increased significantly, in particular, the downward shift of loop6 closed the binding site, resulting in the spatial hindrance of binding. The time proportions of important hydrogen bonds at the binding site decreased sharply, indicating that serious disruption of hydrogen bonds should be the main interaction factor for conformational changes. The residues contributing energetically to binding were also revealed to be in the highly flexible region, which inevitably leads to the decrease in the activity stability at high temperature. These findings provide directions for the modification of thermal stability and perspectives on the research of proteins without experimental structures.

Pleiotropic Modulation of Chitooligosaccharides on Inflammatory Signaling in LPS-Induced Macrophages.

Chitooligosaccharide (COS) is a green and non-toxic cationic carbohydrate that has attracted wide attention in recent years due to its anti-inflammatory activity. However, the anti-inflammatory mechanism of COS remains unclear. In this study, RNA-seq was used to investigate the integrated response of COS to LPS-induced damage in macrophages. The results showed that the experimental group with COS had 2570 genes with significant differences compared to the model group, and that these genes were more enriched in inflammatory and immune pathways. The KEGG results showed that COS induces the pleiotropic modulation of classical inflammatory pathways, such as the Toll-like receptor signaling pathway, NF-κB, MAPK, etc. Based on the RNA-seq data and the RT-qPCR, as well as the WB validation, COS can significantly upregulate the expression of membrane receptors, such as Tlr4, Tlr5, and MR, and significantly inhibits the phosphorylation of several important proteins, such as IκB and JNK. Overall, this study offers deep insights into the anti-inflammatory mechanism and lays the foundation for the early application of COS as an anti-inflammatory drug.

Jellyfish Peptide as an Alternative Source of Antioxidant.

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.

Discovery of a novel jellyfish venom metalloproteinase inhibitor from secondary metabolites isolated from jellyfish-derived fungus Aspergillus versicolor SmT07.

The major jellyfish stings that occur in China are caused by scyphozoan Nemopilema nomurai, whose venom exhibits significant metalloproteinase activity that contributes to the toxic effects of jellyfish envenomation. Researching effective inhibitors suppressing the metalloproteinase activity of jellyfish venom represents a new attempt to cure jellyfish envenomations. In the present study, secondary metabolites produced by the jellyfish-associated fungus Aspergillus versicolor SmT07 were isolated and evaluated for their anti-proteolytic activities. Two xanthones, sterigmatocystin (JC-01) and oxisterigmatocystin C (JC-06), and four alkaloids, cottoquinazoline A (JC-02), phenazine-1-carboxylic acid (JC-03), viridicatin (JC-04) and viridicatol (JC-05), were isolated and identified. Only phenazine-1-carboxylic acid (PCA) showed significant anti-proteolytic activity of jellyfish venom assayed on azocasein, and the IC50 value was 2.16 mM. PCA also significantly inhibited fibrinogenolytic activity, protecting the Bß chain of fibrinogen from degradation when preincubated with jellyfish venom at a ratio of >1:0.6 (PCA:venom, w/w). Molecular docking with several well-characterized snake venom metalloproteinases suggested the venom metalloproteinases inhibitory property of PCA by forming complex interactions with the active site via hydrogen bonds, π-π stacking and salt bridges, which was distinct from the binding mode of batimastat. The present study represents the first study identifying natural jellyfish venom metalloproteinase inhibitors from marine natural products, which may provide an alternative to develop therapeutic agents for treating jellyfish envenomations.

The synthesis, characterization and immunological activity of mucopolysaccharide-quaternized chitosan nanoparticles.

In this study, nanoparticles were prepared by using positively charged quaternized chitosan and negatively charged mucopolysaccharide such as chondroitin sulfate, heparin and hyaluronic acid. The nanoparticles have a stable nanostructure with particle size in 336.2-424.5 nm, potential in 18.5-31.1 mV and polydispersity index PDI of 0.172-0.335. Moreover, their encapsulation efficiency was 68.77 % and 64.89 %, and they have low endotoxin and good stability. It can significantly promote the expression of IL-6, TNF-α, and IL-1ß of DCS cells. Moreover, the in vivo immune activity of heparin-quaternized chitosan-OVA nanoparticles against BALB/C mice was showed that, the nanoparticles could significantly promote the secretion of immunoglobulins in mice including IgG1 and IgG2. And nanoparticle also can promote the production of immune factors. Meanwhile, the expression of immune factor genes was also elevated. Furthermore, the results of tissue section experiments showed that the nanoparticles are safety of the body.

Hepatoprotective Effect of Oyster Peptide on Alcohol-Induced Liver Disease in Mice.

Alcohol-induced liver disease (ALD) has become one of the major global health problems, and the aim of this study was to investigate the characterization of the structure as well as the hepatoprotective effect and mechanism of oyster peptide (OP, MW < 3500 Da) on ALD in a mouse model. The results demonstrate that ethanol administration could increase the activities of aspartate aminotransferase (AST), alanine transaminase (ALT), γ-Glutamyl transferase (GGT), reactive oxygen species (ROS), malondialdehyde (MDA), and triglycerides (TG), as well as increase the interleukin-1ß (IL-1ß), interleukin-6 (IL-6), and tumor necrosis factor (TNF-α) levels (p < 0.01), and reduce the activity of superoxide dismutase (SOD) and the concentration of glutathione (GSH). Those changes were significantly reversed by the application of different doses of OP. Furthermore, the mRNA expressions of nuclear factor elythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and quinone oxidoreductase1 (NQO1) were significantly up-regulated in OP groups, and the mRNA expressions of nuclear factor kappa-light chain enhancer of B cells (NF-κB), TNF-α, and IL-6 were markedly reduced in OP groups compared to that of the model group. Thus, OP had a significant protective effect on ALD through the enhancement of the in vivo antioxidant ability and the inhibition of the inflammatory response as possible mechanisms of action, which therefore suggests that OP might be useful as a natural source to protect the liver from alcohol damage.

The Effect of N-Acetylation on the Anti-Inflammatory Activity of Chitooligosaccharides and Its Potential for Relieving Endotoxemia.

Endotoxemia is mainly caused by a massive burst of inflammatory cytokines as a result of lipopolysaccharide (LPS) invasion. Chitooligosaccharides (COS) is expected to be a potential drug for relieving endotoxemia due to its anti-inflammatory properties. However, the structural parameters of COS are often ambiguous, and the effect of degree of acetylation (DA) of COS on its anti-inflammatory remains unknown. In this study, four COSs with different DAs (0%, 12%, 50% and 85%) and the same oligomers distribution were successfully obtained. Their structures were confirmed by 1H NMR and MS analysis. Then, the effect of DA on the anti-inflammatory activity and relieving endotoxemia potential of COS was researched. The results revealed that COS with a DA of 12% had better anti-inflammatory activity than COSs with other DAs, mainly in inhibiting LPS-induced inflammatory cytokines burst, down-regulating its mRNA expression and reducing phosphorylation of IκBα. Furthermore, this COS showed an obviously protective effect on endotoxemia mice, such as inhibiting the increase in inflammatory cytokines and transaminases, alleviating the injury of liver and intestinal tissue. This study explored the effect of DA on the anti-inflammatory activity of COS for the first time and lays the foundation for the development of COS as an anti-inflammatory drug against endotoxemia.

Design, Synthesis, and Antifungal Activities of Hymexazol Glycosides Based on a Biomimetic Strategy.

Hymexazol (HYM) is irreplaceable for treating soil-borne diseases due to its high efficiency and low cost, as a broad-spectrum fungicide. However, when HYM is absorbed by plants, it is rapidly converted into two glycoside metabolites, and the antifungal activities of these glycosides are inferior to that of HYM. Therefore, in this study, to maintain strong antifungal activity in vitro and in vivo, HYM was glycosylated with amino sugars that have diverse biological activities to simulate plant glycosylation. The antifungal experiment proved that glycoside 15 has the highest antifungal activity, and N-acetyl glucosamine and HYM had obvious synergistic effects. According to the structure-activity relationship studies, glycoside 15 had greater numbers of active electron-rich regions and front-line orbital electrons due to the introduction of N-acetyl glucosamine. Moreover, glycoside 15 can significantly promote plant growth and induce an increase in plant defense enzyme activity. Additionally, compared to HYM, the results of electron microscopy and proteomics revealed that glycoside 15 has a unique antifungal mechanism. The promising antifungal activity and interactions with plants mean that glycoside 15 is a potential green fungicide candidate. Furthermore, this research conducted an interesting exploration of the agricultural applications of amino sugars.

Jellyfish Nemopilema nomurai causes myotoxicity through the metalloprotease component of venom.

Jellyfish envenomation is a common medical problem in many countries. However, the myotoxicity and effector molecules of scyphozoan venoms remain uninvestigated. Here, we present the myotoxicity of nematocyst venom from Nemopilema nomurai (NnNV), a giant venomous scyphozoan from China, for the first time, using in vivo models with inhibitors. NnNV was able to induce remarkable myotoxicity including significant muscle swelling, increasing the content of CK and LDH in serum, stimulating inflammation of muscle tissue, and destroying the structure of muscle tissue. In addition, the metalloproteinase inhibitors BMT and EDTA significantly reduced the myotoxicity induced by NnNV. Moreover, BMT and EDTA could decrease the inflammatory stimulation and necrosis of muscle tissue caused by the venom. These observations suggest that the metalloproteinase components of NnNV make a considerable contribution to myotoxicity. This study contributes to understanding the effector molecules of muscle injury caused by jellyfish stings and suggests a new idea for the treatment of scyphozoan envenomation.