Experiments showed the agglutinin activity in the hemolymph of both spring wild parents and autumncultured individuals of the shrimp Penaeus chinensis Osbeck with the body length of 12 - 16 cm as all 32. Thehemolymph ...Experiments showed the agglutinin activity in the hemolymph of both spring wild parents and autumncultured individuals of the shrimp Penaeus chinensis Osbeck with the body length of 12 - 16 cm as all 32. Thehemolymph of the above shrimps showed agglutinant reaction to all A-, B- and O-type human erythrocytes, with thereaction titer equal to that of rabbit blood cells. However, the agglutinin activity level was enhanced after being in-duced by Vibro, which was 128 after 24 h of induction and reached the peak value 256 after 48 h of induction, thendecreased gradually.展开更多
Knowledge of dietary compositions is essential to the recovery of the dwindling populations of horseshoe crabs in China.The feeding habits of Tachypleus tridentatus and Carcinoscorpis rotundicauda of Pearl Bay in sout...Knowledge of dietary compositions is essential to the recovery of the dwindling populations of horseshoe crabs in China.The feeding habits of Tachypleus tridentatus and Carcinoscorpis rotundicauda of Pearl Bay in southwestern China were studied by stable isotope analysis.Hemolymph samples of the two species were obtained from two age groups living in different habitats(inner,central,and outer Bay).In addition,their potential food source samples were collected in May 2019.Results showed that the mean values ofδ^(13)C andδ^(15)N in C.rotundicauda individuals ranged from−19.01‰to−16.47‰and from 10.49‰to 13.5‰,respectively,and those of T.tridentatus ranged from−19.12‰to−14.96‰and from 8.78‰to 13.48‰,respectively.These values indicated that the horseshoe crabs have a wide variety of food sources and therefore are highly omnivorous.No remarkably correlations were found betweenδ^(13)C andδ^(15)N values and individual widths in the two species.Thirteen potential food sources were selected,in which Sipunculus nudus,Batillaria zonalisi,and Ulva lactuca contributed largely to the dietary compositions of the two species of horseshoe crabs.No significant correlation was found between the feeding habits and habitat geochemical characteristics of the horseshoe crabs.All these results possibly indicate the high diversity of their food sources and the indiscriminate dietary compositions of the two Asian horseshoe crabs.展开更多
This study investigated the effects of silkworm hemolymph-derived storage protein 2 (SP2) on the whitening process in mouse B16F1 melanoma cells. After the cells were treated with various concentrations of SP2 (0.1 - ...This study investigated the effects of silkworm hemolymph-derived storage protein 2 (SP2) on the whitening process in mouse B16F1 melanoma cells. After the cells were treated with various concentrations of SP2 (0.1 - 1.0 mg/mL), cytotoxicity, melanin contents, and differences in mRNA and protein expression associated with melanogenesis were observed. No cytotoxicity was observed when cells were treated with SP2, even with increased SP2 concentrations of up to 2.0 mg/mL. When treated with various SP2 concentrations in the cells, the protein and mRNA expression of tyrosinase were dose-dependently decreased, respectively, and inhibition of tyrosinase was further increased by 50.0% with increasing SP2 concentration of 1.0 mg/mL. Expression mRNAs coding tyrosinase related protein-1 and protein-2 (TRP-1 and TRP-2) was also significantly decreased in a dose dependent manner. When measuring the melanin content in melanoma cells, SP2 at 1 mg/mL inhibited melanin synthesis by 73.5% compared with non-treated cells. The inhibitory effect was 2.8-fold higher than that obtained using arbutin as a positive control. This study demonstrates that SP2, as a whitening material, is capable of suppressing melanin synthesis through the downregulation of proteins and genes in the melanin biosynthesis pathway.展开更多
Insects are well recognized as a source of potentially useful compounds for modern medicine. Development of anticancer drugs from natural resources has been performed throughout the world. In the present study, antica...Insects are well recognized as a source of potentially useful compounds for modern medicine. Development of anticancer drugs from natural resources has been performed throughout the world. In the present study, anticancer activity of the hemolymph and fat body of Sarcophaga argyrostoma third larval instars is assayed against human breast cancer cell line (MDA-MB-231 cells). The cytotoxicity of the hemolymph and fat body samples were determined. The results showed that growth of MDA-MB-231 cells was inhibited at different concentrations upon 24 h of exposure. There is no inhibitory activity against Vero cells under these experimental conditions. Protein profile of the hemolymph and fat body were extracted and separated using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Protein analysis showed differences in number of electrophoretic protein bands with different molecular weights between treated and control larvae. The present work assumes that hemolymph and fat body tissue have cytotoxicity against MDA-MB-231 cells and these results exhibit that larvae from S. argyrostoma could be used as a good source for developing anti-cancer agents and knowledge of these anti-cancer compounds will lead to better control of human disease.展开更多
Peptides in shrimp hemolymph play an important role in the innate immune response.Analysis of hemolymph will help to detect and identify potential novel biomarkers of microbial infection.We used magnetic bead-based pu...Peptides in shrimp hemolymph play an important role in the innate immune response.Analysis of hemolymph will help to detect and identify potential novel biomarkers of microbial infection.We used magnetic bead-based purification(ClinProt system) and matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS) to characterize shrimp hemolymph peptides.Shrimp serum and plasma were used as the source of samples for comparative analysis,and it was found that serum was more suitable for shrimp hemolymph peptidomic analysis.To screen potential specific biomarkers in serum of immune-challenged shrimps,we applied magnetic bead-based MALDI-TOF MS to serum samples from 10 immune-challenged and 10 healthy shrimps.The spectra were analyzed using FlexAnalysis 3.0 and ClinProTools 2.1 software.Thirteen peptide peaks significantly different between the two groups were selected as candidate biomarkers of lipopolysaccharide(LPS)-infection.The diagnostic model established by genetic algorithm using five of these peaks was able to discriminate LPS-challenged shrimps from healthy control shrimps with a sensitivity of 90% and a specificity of 100%.Our approach in MALDITOF MS-based peptidomics is a powerful tool for screening bioactive peptides or biomarkers derived from hemolymph,and will help to enable a better understanding of the innate immune response of shrimps.展开更多
Two-dimensional gel electrophoresis （2-DE） followed by matrix-assisted laser desorption ionization- time-of-flight/time-of-flight mass spectrometry （MS） analysis were used to charaterize the hemolymph proteomic pr...Two-dimensional gel electrophoresis （2-DE） followed by matrix-assisted laser desorption ionization- time-of-flight/time-of-flight mass spectrometry （MS） analysis were used to charaterize the hemolymph proteomic profiles of the silkworm, Bombyx mori. At days 4 （V4） and 5 （VS） of the fifth （final） instar, when the larvae were at the fast-growing stage, we found dramatic changes in spots representing proteins having an approximate molecular weight （MW） of 30 kDa. Of these spots, four 30K proteins were highly up- regulated, implying a close association with the growth and development ofB. mori larvae. To understand the molecular basis and underlying mechanisms involved in development and metamorphosis, the proteome of whole hemolymph at V5 was analyzed using shotgun liquid chromatography tandem mass spectrometry with an LTQ-Orbitrap. A total of 108 proteins were identified without any false discovery hits. These proteins were involved in a variety of cellular functions, including metabolism, development, nutrient transport and reserve, and defense response. Gene ontology analysis showed that 3.4% of these proteins had nutrient reservoir activities and 5.7% were involved in the response to stimulus. Pathway analysis revealed that 22 proteins with common targets were involved in various cellular processes such as immunity, differentiation, proliferation and metamorphosis. These results suggested that some key factors such as the 30K proteins in hemolymph play important roles in B. mori growth and development. Moreover, the multiple functions of hemolymph may be operated by a complex biological network.展开更多
During insect larval-pupal metamorphosis,proteins in the hemolymph are absorbed by the fat body for the maintenance of intracellular homeostasis;however,the type of proteins and how these proteins are internalized int...During insect larval-pupal metamorphosis,proteins in the hemolymph are absorbed by the fat body for the maintenance of intracellular homeostasis;however,the type of proteins and how these proteins are internalized into the fat body are unclear.In Bombyx mori,the developmental profiles o f total proteins in the hemolymph and fat body showed that hemolymph-decreased protein bands(55-100 kDa)were in accordance with those protein bands that increased in the fat body.Inhibition o f clathrin-dependent endocytosis predominantly blocked the transportation o f 55-100 kDa proteins from the hemolymph into the fat body,which was further verified by RNA interference treatment o f Bmclathrin.Six hexamerins were shown to comprise〜90%of the total identified proteins in both the hemolymph and fat body by mass spectrum(MS)analysis.In addition,hemolymphspecific proteins were mainly involved in material transportation,while fat body-specific proteins particularly participated in metabolism.In this paper,four hexamerins were found for the first time,and potential proteins absorbed by the fat body from the hemolymph through clathrin-dependent endocytosis were identified.This study sheds light on the protein absorption mechanism during insect metamorphosis.展开更多
Acute-phase response is documented to be a significant mechanism of innate immunity in vertebrates and invertebrates.In this study,proteomic methodologies were applied for different protein expressions in hemolymph of...Acute-phase response is documented to be a significant mechanism of innate immunity in vertebrates and invertebrates.In this study,proteomic methodologies were applied for different protein expressions in hemolymph of Scylla serrata challenged by Vibrio parahaemolyticus after immunization,and in muscles of the crabs separately chal-lenged by V.parahaemolyticus,V.anguillarum and Aeromonas hydrophila.Up-regulated cryptocyanin is documented in the hemolymph and up-regulated calexcitin,wingless(fragment)and tachykinin-related peptide in the muscle as acute-phase proteins.All the four altered proteins were responsible for bacterial stress,but cryptocyanin seemed to be a memory response protein against the chal-lenge by a live bacterium after immunization of the live cells.These up-regulated proteins can be indicative of an understanding of immunity of a crab.展开更多
There is an increasing interest in discovering new antibacterial agents derived from nature to enhance the treatment of various bacterial infections.Defensins and their derived peptide fragments exhibit significant an...There is an increasing interest in discovering new antibacterial agents derived from nature to enhance the treatment of various bacterial infections.Defensins and their derived peptide fragments exhibit significant antibacterial activity without any cytotoxic effects,making them attractive features for potential novel antibacterial therapeutics.Crassostrea gigas,a traditional seafood that has been used worldwide for centuries,has its shells applied in Chinese medicine as Ostreae concha.In this study,bioinformatics analysis was used to obtain a novel antibacterial peptide,CGD-1,derived from marine Chinese medicine Ostreae concha.The physicochemical characterization and circular dichroism analysis results demonstrated that CGD-1 assembled into anα-helical structure in a simulated membrane environment,and it displayed antibacterial action against Gram-negative bacteria.The minimal inhibitory concentrations against both Pseudomonas aeruginosa ATCC27853 and Escherichia coli ATCC25922 were 25μM.CGD-1 was able to efficiently permeate the cell membrane.Changes in bacterial cell morphology were evaluated using a field emission scanning electron microscope.The results suggested that CGD-1 exerted its antibacterial activity through permeabilizing and disrupting the bacterial cell membrane.Therefore,CGD-1 may have potential applications in fighting against pathogenic bacteria such as P.aeruginosa and E.coli.展开更多
文摘Experiments showed the agglutinin activity in the hemolymph of both spring wild parents and autumncultured individuals of the shrimp Penaeus chinensis Osbeck with the body length of 12 - 16 cm as all 32. Thehemolymph of the above shrimps showed agglutinant reaction to all A-, B- and O-type human erythrocytes, with thereaction titer equal to that of rabbit blood cells. However, the agglutinin activity level was enhanced after being in-duced by Vibro, which was 128 after 24 h of induction and reached the peak value 256 after 48 h of induction, thendecreased gradually.
基金the Scientific Research Project of Huaqiao University(No.605-50X18005)Guangxi Bagui Youth Scholars ProgrammeGuangxi Recruitment Program of 100 Global Experts。
文摘Knowledge of dietary compositions is essential to the recovery of the dwindling populations of horseshoe crabs in China.The feeding habits of Tachypleus tridentatus and Carcinoscorpis rotundicauda of Pearl Bay in southwestern China were studied by stable isotope analysis.Hemolymph samples of the two species were obtained from two age groups living in different habitats(inner,central,and outer Bay).In addition,their potential food source samples were collected in May 2019.Results showed that the mean values ofδ^(13)C andδ^(15)N in C.rotundicauda individuals ranged from−19.01‰to−16.47‰and from 10.49‰to 13.5‰,respectively,and those of T.tridentatus ranged from−19.12‰to−14.96‰and from 8.78‰to 13.48‰,respectively.These values indicated that the horseshoe crabs have a wide variety of food sources and therefore are highly omnivorous.No remarkably correlations were found betweenδ^(13)C andδ^(15)N values and individual widths in the two species.Thirteen potential food sources were selected,in which Sipunculus nudus,Batillaria zonalisi,and Ulva lactuca contributed largely to the dietary compositions of the two species of horseshoe crabs.No significant correlation was found between the feeding habits and habitat geochemical characteristics of the horseshoe crabs.All these results possibly indicate the high diversity of their food sources and the indiscriminate dietary compositions of the two Asian horseshoe crabs.
文摘This study investigated the effects of silkworm hemolymph-derived storage protein 2 (SP2) on the whitening process in mouse B16F1 melanoma cells. After the cells were treated with various concentrations of SP2 (0.1 - 1.0 mg/mL), cytotoxicity, melanin contents, and differences in mRNA and protein expression associated with melanogenesis were observed. No cytotoxicity was observed when cells were treated with SP2, even with increased SP2 concentrations of up to 2.0 mg/mL. When treated with various SP2 concentrations in the cells, the protein and mRNA expression of tyrosinase were dose-dependently decreased, respectively, and inhibition of tyrosinase was further increased by 50.0% with increasing SP2 concentration of 1.0 mg/mL. Expression mRNAs coding tyrosinase related protein-1 and protein-2 (TRP-1 and TRP-2) was also significantly decreased in a dose dependent manner. When measuring the melanin content in melanoma cells, SP2 at 1 mg/mL inhibited melanin synthesis by 73.5% compared with non-treated cells. The inhibitory effect was 2.8-fold higher than that obtained using arbutin as a positive control. This study demonstrates that SP2, as a whitening material, is capable of suppressing melanin synthesis through the downregulation of proteins and genes in the melanin biosynthesis pathway.
文摘Insects are well recognized as a source of potentially useful compounds for modern medicine. Development of anticancer drugs from natural resources has been performed throughout the world. In the present study, anticancer activity of the hemolymph and fat body of Sarcophaga argyrostoma third larval instars is assayed against human breast cancer cell line (MDA-MB-231 cells). The cytotoxicity of the hemolymph and fat body samples were determined. The results showed that growth of MDA-MB-231 cells was inhibited at different concentrations upon 24 h of exposure. There is no inhibitory activity against Vero cells under these experimental conditions. Protein profile of the hemolymph and fat body were extracted and separated using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Protein analysis showed differences in number of electrophoretic protein bands with different molecular weights between treated and control larvae. The present work assumes that hemolymph and fat body tissue have cytotoxicity against MDA-MB-231 cells and these results exhibit that larvae from S. argyrostoma could be used as a good source for developing anti-cancer agents and knowledge of these anti-cancer compounds will lead to better control of human disease.
基金Supported by the National Natural Science Foundation of China(No.30600458)
文摘Peptides in shrimp hemolymph play an important role in the innate immune response.Analysis of hemolymph will help to detect and identify potential novel biomarkers of microbial infection.We used magnetic bead-based purification(ClinProt system) and matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS) to characterize shrimp hemolymph peptides.Shrimp serum and plasma were used as the source of samples for comparative analysis,and it was found that serum was more suitable for shrimp hemolymph peptidomic analysis.To screen potential specific biomarkers in serum of immune-challenged shrimps,we applied magnetic bead-based MALDI-TOF MS to serum samples from 10 immune-challenged and 10 healthy shrimps.The spectra were analyzed using FlexAnalysis 3.0 and ClinProTools 2.1 software.Thirteen peptide peaks significantly different between the two groups were selected as candidate biomarkers of lipopolysaccharide(LPS)-infection.The diagnostic model established by genetic algorithm using five of these peaks was able to discriminate LPS-challenged shrimps from healthy control shrimps with a sensitivity of 90% and a specificity of 100%.Our approach in MALDITOF MS-based peptidomics is a powerful tool for screening bioactive peptides or biomarkers derived from hemolymph,and will help to enable a better understanding of the innate immune response of shrimps.
基金Supported by the National Natural Science Foundation of China （No.30170742） and National Key Foundational Research Project of China （No.G1999012012）.
文摘Two-dimensional gel electrophoresis （2-DE） followed by matrix-assisted laser desorption ionization- time-of-flight/time-of-flight mass spectrometry （MS） analysis were used to charaterize the hemolymph proteomic profiles of the silkworm, Bombyx mori. At days 4 （V4） and 5 （VS） of the fifth （final） instar, when the larvae were at the fast-growing stage, we found dramatic changes in spots representing proteins having an approximate molecular weight （MW） of 30 kDa. Of these spots, four 30K proteins were highly up- regulated, implying a close association with the growth and development ofB. mori larvae. To understand the molecular basis and underlying mechanisms involved in development and metamorphosis, the proteome of whole hemolymph at V5 was analyzed using shotgun liquid chromatography tandem mass spectrometry with an LTQ-Orbitrap. A total of 108 proteins were identified without any false discovery hits. These proteins were involved in a variety of cellular functions, including metabolism, development, nutrient transport and reserve, and defense response. Gene ontology analysis showed that 3.4% of these proteins had nutrient reservoir activities and 5.7% were involved in the response to stimulus. Pathway analysis revealed that 22 proteins with common targets were involved in various cellular processes such as immunity, differentiation, proliferation and metamorphosis. These results suggested that some key factors such as the 30K proteins in hemolymph play important roles in B. mori growth and development. Moreover, the multiple functions of hemolymph may be operated by a complex biological network.
基金This study was supported by the National Natu-ral Science Foundation of China(grants 31472042 and 31672368 to LT,31702053 to KL and 31702065 to HY)the Natural Science Foundation of Guangdong Province(grant 2017A030311024 to LT)。
文摘During insect larval-pupal metamorphosis,proteins in the hemolymph are absorbed by the fat body for the maintenance of intracellular homeostasis;however,the type of proteins and how these proteins are internalized into the fat body are unclear.In Bombyx mori,the developmental profiles o f total proteins in the hemolymph and fat body showed that hemolymph-decreased protein bands(55-100 kDa)were in accordance with those protein bands that increased in the fat body.Inhibition o f clathrin-dependent endocytosis predominantly blocked the transportation o f 55-100 kDa proteins from the hemolymph into the fat body,which was further verified by RNA interference treatment o f Bmclathrin.Six hexamerins were shown to comprise〜90%of the total identified proteins in both the hemolymph and fat body by mass spectrum(MS)analysis.In addition,hemolymphspecific proteins were mainly involved in material transportation,while fat body-specific proteins particularly participated in metabolism.In this paper,four hexamerins were found for the first time,and potential proteins absorbed by the fat body from the hemolymph through clathrin-dependent endocytosis were identified.This study sheds light on the protein absorption mechanism during insect metamorphosis.
基金supported by Hi-Tech Research and Development Program of China (No.2002AA603013)the Natural Science Foundation of Fujian Province (No.B0110005).
文摘Acute-phase response is documented to be a significant mechanism of innate immunity in vertebrates and invertebrates.In this study,proteomic methodologies were applied for different protein expressions in hemolymph of Scylla serrata challenged by Vibrio parahaemolyticus after immunization,and in muscles of the crabs separately chal-lenged by V.parahaemolyticus,V.anguillarum and Aeromonas hydrophila.Up-regulated cryptocyanin is documented in the hemolymph and up-regulated calexcitin,wingless(fragment)and tachykinin-related peptide in the muscle as acute-phase proteins.All the four altered proteins were responsible for bacterial stress,but cryptocyanin seemed to be a memory response protein against the chal-lenge by a live bacterium after immunization of the live cells.These up-regulated proteins can be indicative of an understanding of immunity of a crab.
基金This research work was financially supported National Natural Science Foundation of China(No.82003910)Guangdong Basic and Applied Basic Research Foundation(2020A1515110550)+2 种基金Funding Key R&D Program of Shandong Province(2022SFGC0105)Exploration Innovation Team(2021GXRC062),Jinan Talent Project for Universities(202228088)Key innovation Project of Qilu University of Technology(Shandong Academy of Sciences)(2022JBZ01-06).
文摘There is an increasing interest in discovering new antibacterial agents derived from nature to enhance the treatment of various bacterial infections.Defensins and their derived peptide fragments exhibit significant antibacterial activity without any cytotoxic effects,making them attractive features for potential novel antibacterial therapeutics.Crassostrea gigas,a traditional seafood that has been used worldwide for centuries,has its shells applied in Chinese medicine as Ostreae concha.In this study,bioinformatics analysis was used to obtain a novel antibacterial peptide,CGD-1,derived from marine Chinese medicine Ostreae concha.The physicochemical characterization and circular dichroism analysis results demonstrated that CGD-1 assembled into anα-helical structure in a simulated membrane environment,and it displayed antibacterial action against Gram-negative bacteria.The minimal inhibitory concentrations against both Pseudomonas aeruginosa ATCC27853 and Escherichia coli ATCC25922 were 25μM.CGD-1 was able to efficiently permeate the cell membrane.Changes in bacterial cell morphology were evaluated using a field emission scanning electron microscope.The results suggested that CGD-1 exerted its antibacterial activity through permeabilizing and disrupting the bacterial cell membrane.Therefore,CGD-1 may have potential applications in fighting against pathogenic bacteria such as P.aeruginosa and E.coli.
