Tissue culture is one of the most promising and practical methods for conserving endangered plant species.Therefore,the present study evaluates the conservation of the endangered seaweed Sargassum fusiforme through ti...Tissue culture is one of the most promising and practical methods for conserving endangered plant species.Therefore,the present study evaluates the conservation of the endangered seaweed Sargassum fusiforme through tissue culturing techniques from different explants for the first time.Besides,the genetic variation of the mother plant and cultivated explants using inter-simple sequence repeat(ISSR)techniques;this methodology was the first to be illustrated in such work for algae tissue culturing.The regeneration results have shown that different explants could induce shoot and rhizoid morphogenesis with a total number of blades of 2436,1011,1466,678,and 6 from apical parts,stipe with blades,two-segmented seedlings,rhizoids,and stipe without blades,respectively.The total length was 234,181.8,83.5,and 81.8 cm from the two-segmented seedlings,apical parts,stipe without blades,and stipe with blades,respectively.At the same time,the total wet weight was 73.148,48.369,35.731,18.588,and 2.035 g from the apical parts,the two-segmented seedlings,stipe with blades,rhizoids,and stipe without blades,respectively.Micropropagation of S.fusiforme was successfully achieved with apical,stipe,and stolon segments using free PES media.It is suggested that the applied genetic fingerprint is valid for S.fusiforme and will respond well to molecular marker assistance in cultivation.The significance of S.fusiforme and its exposure to being endangered due to over-exploitation have made its regeneration in vitro a subject of interest in this study.Thus,this report represents the successful regeneration of S.fusiforme and explores the genetic uniformity or somaclonal variation of the obtained seedlings using the ISSR-PCR marker for the first time.展开更多
[Objectives]This study was conducted to investigate the application of Sargassum fusiforme(Harv.)Setch.in cigarettes.[Methods]Tobacco-specific nitrosamines in the smoke of cigarettes added with ethanol extract of S.fu...[Objectives]This study was conducted to investigate the application of Sargassum fusiforme(Harv.)Setch.in cigarettes.[Methods]Tobacco-specific nitrosamines in the smoke of cigarettes added with ethanol extract of S.fusiforme were determined,and the compounds related to the aroma of S.fusiforme were identified by flavor-smelling experiment.[Results]With the addition of the ethanol extract of S.fusiforme,the decrease in the total amount of four tobacco-specific nitrosamines in mainstream cigarette smoke reached 16.42%.The results of the flavor-smelling experiment showed that the aroma of S.fusiforme might be related to(R)-5,6,7,7A-tetrahydro-4,4,7A-trimethyl-2(4H)-benzofuranone,glycerol,ethyl palmitate,methyl palmitate,ethyl linoleate,methyl(Z,Z,Z)-9,12,15-octadecatrienoate,ethyl(Z,Z,Z)-9,12,15-octadecatrienoate,phytol,and tetradecanoic acid.[Conclusions]The ethanol extract of S.fusiforme has the potential function of reducing the content of tobacco-specific nitrosamines in smoke and improving the taste of cigarettes.展开更多
This study aimed to investigate the effects of Sargassum fusiforme polysaccharide(SFPS I,II,and III)on the apoptosis and regulation of human erythroleukemia(HEL)cells.The effect of different doses of SFPS on HEL cell ...This study aimed to investigate the effects of Sargassum fusiforme polysaccharide(SFPS I,II,and III)on the apoptosis and regulation of human erythroleukemia(HEL)cells.The effect of different doses of SFPS on HEL cell growth was detected using the Cell Counting Kit-8 method,and apoptosis was detected by Hoechst staining.Cell cycle distribution and apoptosis were detected using flow cytometry.Expression of the cell cycle gene,p53,antiapoptotic genes,Bcl-xL and Bcl-2,and pro-apoptotic genes,Bax,Bad,and Caspase-3,as well as the expression of the corresponding proteins,were detected using real-time quantitative polymerase chain reaction(qPCR)and Western blot.The results showed that SFPS Ⅱ and Ⅲ decreased HEL cell viability and induced HEL cell apoptosis.Different concentrations of SFPS(Ⅰ,Ⅱ,and Ⅲ)were detected that induced much less toxic effect in normal human embryonic lung(MRC-5)cells,and SFPS Ⅰ increased cell proliferation,indicating its favorable selectivity towards cancer cells.The mechanism by which SFPS induced apoptosis was also found to be related to the induction of cell cycle arrest in the G0/G1 phase and the increased expression of apoptosis-related genes and proteins.We concluded that SFPS induces HEL cell apoptosis,possibly via activation of the Caspase pathway,providing the theoretical basis for the development of SFPS-based anti-tumor drug products.展开更多
Heparin has been used as an anticoagulant drug for many years, but it has significant side ef fects. In the search for good substitutes, low molecular weight(MW) polysaccharides from S argassum fusiforme have been exa...Heparin has been used as an anticoagulant drug for many years, but it has significant side ef fects. In the search for good substitutes, low molecular weight(MW) polysaccharides from S argassum fusiforme have been examined and confi rmed to possess biological activities. Here, S. fusiforme polysaccharides(SFP) were extracted and subjected to a hydrogen peroxide(H_2O_2) oxidation method for the preparation of low-MW SFP(LSFP). The effects of temperature, pH, and H_2O_2 concentration on the degradation process were also examined. Several LSFP of 36, 9, 5.7, and 2.7 kDa were obtained under different conditions, and their anticoagulant activities studied in vitro. The results showed that SFP and LSFP prolonged activated partial thromboplastin(APTT), prothrombin(PT) and thrombin times(TT) significantly, indicating that these low MW polysaccharides possessed anticoagulant activity in the intrinsic, extrinsic, and common coagulation pathways. As these ef fects were related to the MW of the polysaccharides in APTT and TT but not in PT, the contents of the monosaccharide fucose and sulfate and the polysaccharide MW could have exerted combined ef fects. The details of this mechanism require further verifi cation.展开更多
Since the 1980s,Sargassum fusiforme has been cultivated in Zhejiang,South China,and nowadays it becomes one of the important commercial seaweeds in China.With traditions of eating habits in the East Asian countries,th...Since the 1980s,Sargassum fusiforme has been cultivated in Zhejiang,South China,and nowadays it becomes one of the important commercial seaweeds in China.With traditions of eating habits in the East Asian countries,this brown alga is used as food,because it contained functional oligo/polysaccharides and chemical components,and was regarded playing roles in antioxidant activities and regulating immunology.Through over 15 years’selection,breeding and cultivation,we obtained three strains with good traits and testified their characters during the production,which included the cultivars with high yield and other two good characters,either all the selected strains were applied in the Sargassum production.To avoid confusion during the selection and nursery,it was preferred to establish one fingerprint for distinguishing the Sargassum cultivars from different strains.Random amplified polymorphic DNA(RAPD)and inter-simple sequence repeat(ISSR)methods were adopted to analyze the genetic diversities of the selected S.fusiforme strains.With that,one fingerprint with RAPD markers was constructed,and one sequence characterized amplifi ed region(SCAR)marker to S.fusiforme was obtained.It is indicated that the applied fingerprint could be valid in S.fusiforme genetic and germplasm justification,and will be positive to molecular marker assistance in its selection and cultivation.展开更多
Sargassum fusiforme(S.fusiforme)has been used as an ingredient in Chinese herbal medicine for thousands of years.However,there are a limited number of studies concerning its therapeutic mechanism.High performance gel ...Sargassum fusiforme(S.fusiforme)has been used as an ingredient in Chinese herbal medicine for thousands of years.However,there are a limited number of studies concerning its therapeutic mechanism.High performance gel permeation chromatography(HPGPC)analysis showed that the average molecular weight of the S.fusiforme polysaccharide,SFPS 191212,is 43 kDa.SFPS 191212 is composed of mannose,rhamnose,galactose,xylose,glucose,and fucose(at a molar ratio:2.1:2.9:1.8:15.5:4.6:62.5)withα-andβ-configurations.The present research evaluated the anti-tumor potential of the S.fusiforme polysaccharide in human erythroleukemia(HEL)cells in vitro.To explore the SFPS 191212’s apoptosis mechanism in HEL cells,transcriptome analysis was performed on HEL cells that were incubated with SFPS 191212.The inhibitory effect of SFPS 191212 on HEL cell growth was also analyzed.It was found that SFPS 191212 inhibited HEL cell proliferation,reduced cell viability in a concentration-dependent manner,and induced an insignificant toxic effect on normal human embryonic lung(MRC-5)cells.Compared with the control group,transcriptome analysis identified a total of 598 differentially expressed genes(DEGs),including 243 up-regulated genes and 355 downregulated genes.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed on all DEGs,and 900 GO terms and 52 pathways were found to be significantly enriched.Finally,23 DEGs were randomly selected and confirmed by quantitative real-time polymerase chain reaction(qRT-PCR).Moreover,SFPS 191212 down-regulated the PI3K/Akt signal transduction pathway.Our results provide a framework for understanding the effect of SFPS 191212 on cancer cells and can serve as a resource for delineating the anti-tumor mechanisms of S.fusiforme.展开更多
Objective:To investigate whether ethanol extracts of Hizikia fusiforme could induce apoptosis in human prostate cancer PC3 cells.Methods:Cell viability was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetr...Objective:To investigate whether ethanol extracts of Hizikia fusiforme could induce apoptosis in human prostate cancer PC3 cells.Methods:Cell viability was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.Apoptosis and mitochondrial membrane potential(MMP)were measured using flow cytometry in PC3 cells.DNA damage was assessed by nuclear staining and DNA fragmentation assay.Expressions of apoptosis-associated proteins were determined by Western blotting assays.Activities of caspase-3,-8,and-9 were determined by colorimetric assay.Moreover,intracellular reactive oxygen species(ROS)generation was detected using a flow cytometer and fluorescence microscope.Results:Treatment of PC3 cells with ethanol extracts of Hizikia fusiforme inhibited proliferation,which was associated with induction of apoptosis,and accompanied by increased expression of Fas,Fas-ligand(Fas L),Bax and t Bid,and decreased expression of Bcl-2.In addition,ethanol extracts of Hizikia fusiforme reduced c-Flip expression and activated caspase-8,-9 and-3,resulting in an increase in poly(ADP-ribose)polymerase(PARP)cleavage.However,in the presence of a pan-caspase inhibitor,ethanol extracts of Hizikia fusiforme-mediated growth inhibition and apoptosis were significantly attenuated.Ethanol extracts of Hizikia fusiforme also destroyed the integrity of mitochondria due to the loss of MMP,leading to cytosolic release of cytochrome c.Moreover,the levels of ROS were markedly increased by treatment with ethanol extracts of Hizikia fusiforme,which was significantly suppressed by the ROS scavenger N-acetyl-L-cysteine.Further investigation of whether ethanol extracts of Hizikia fusiforme-induced apoptosis was related to the generation of ROS was conducted and the results showed that N-acetyl-L-cysteine fully blocked ethanol extracts of Hizikia fusiforme-induced apoptotic events including loss of MMP,activation of caspase-3,the cytosolic release of cytochrome c and cytotoxicity.Conclusions:Ethanol extracts of Hizikia fusiforme have chemopreventive potential via induction of ROS-dependent apoptosis.Therefore,ethanol extracts of Hizikia fusiforme may be useful for developing effective and selective natural sources to inhibit cancer cell proliferation.展开更多
The response surface methodology(RSM) combined with bioassays was employed to optimize the extraction process of crude fucose-containing sulphated polysaccharides(c FCSP) from Sargassum fusiforme. The central composit...The response surface methodology(RSM) combined with bioassays was employed to optimize the extraction process of crude fucose-containing sulphated polysaccharides(c FCSP) from Sargassum fusiforme. The central composite design(CCD) was used with four variables, five levels, and four responses. The four variables were p H value of hydrochloric acid solution, extraction temperature(℃), ratio of liquid to raw material(m L g^(-1)), and extraction time(h), respectively. Chemical and bioassay indices were used in combination as the response parameters, which included the yield of c FCSP, fucose content, proliferation rate of spleen cells, and lipopolysaccharide-induced proliferation of splenocytes. The experimental data were fitted to a second-order polynomial equation using multiple regression analysis, and examined using the appropriate statistical methods. The best extraction conditions were as follows: the p H value of hydrochloric acid solution was 3.50; the extraction temperature was 100℃; the ratio of liquid to raw material was 15.00 m L g^(-1) and the extraction time was 2.50 h. The experimental yield was close to the predicted from the model. The extract could promote spleen lymphocyte proliferation, especially the lipopolysaccharide-induced lymphocyte proliferation in vitro, which suggested that its immunomodulatory effect on B lymphocytes. Therefore, c FCSP extracted from S. fusiforme could be utilized as an immunostimulant in functional foods and pharmaceutical industry in future.展开更多
[Objectives] The research aimed to optimize extraction process of fucoidan from Sargassum fusiforme( FSF) and study its moisturizing performance. [Methods]Extracting condition of FSF by cellulase hydrolysis-ultrasonic...[Objectives] The research aimed to optimize extraction process of fucoidan from Sargassum fusiforme( FSF) and study its moisturizing performance. [Methods]Extracting condition of FSF by cellulase hydrolysis-ultrasonic assisted extraction method was optimized. The influences of solvent p H,enzyme dosage,extraction temperature,cellulose hydrolysis time,ultrasonication time,and the ratio of material to liquid on FSF were investigated by single factor and orthogonal experiments. [Results] The optimum extraction conditions were as followings:p H,4. 5; enzyme dosage,1%; extraction temperature,40℃; cellulose hydrolysis time,2 h; ultrasonic time,15 min; and the ratio of material to liquid,1∶ 10( g∶ m L). Under the optimal condition,the extraction yield of FSF was 8. 50%,RSD = 2. 74%. The short-time hygroscopicity( within 8 h) of crude extract of fucoidan from S. fusiforme( CEFSF) was better than glycerin,butanediol,and sodium alginate,and the moisture retention capacity of 1% CEFSF aqueous solution was better than 1% butanediol or 1% sodium alginate,and was equal to 5% glycerin under relative humidity of 43% and 81%. The determination results of skin moisture content and transepidermal water loss rate( TEWL)showed that: 5% CEFSF solution had good moisturizing effect. [Conclusions]The research could provide certain reference for deep development of S. fusiforme.展开更多
Cadmium(Cd)is a common heavy metal pollutant in the aquatic environment,generally toxic to plant growth and leading to growth inhibition and biomass reduction.To study the oxidation resistance in Sargassum fusiforme s...Cadmium(Cd)is a common heavy metal pollutant in the aquatic environment,generally toxic to plant growth and leading to growth inhibition and biomass reduction.To study the oxidation resistance in Sargassum fusiforme seedlings in response to inorganic Cd stress,we cultured the seedlings under two different Cd levels:natural seawater and high Cd stress.High Cd stress significantly inhibited the seedlings growth,and darkened the thalli color.Additionally,the pigment contents,growth rate,peroxidase(POD)activity,dehydroascorbic acid(DHA)content,and glutathione reductase(GR)activity in S.fusiforme were significantly reduced by high Cd treatment.Contrarily,the Cd accumulation,Cd2+absorption rate,dark respiration/net photosynthetic rate(Rd/Pn),ascorbic acid(Vc)content,soluble protein(SP)content,glutathione(GSH),and the activities of superoxide dismutase(SOD)and catalase(CAT)of S.fusiforme under Cd treatment significantly increased compared to the control group.The decrease of malondialdehyde(MDA)was not significant.Although S.fusiforme seedlings increased the antioxidant activities of POD,SOD,Vc,and the AsA-GSH cycle to disseminate H2O2 and maintain healthy metabolism,high Cd stress caused Cd accumulation in the stem and leaves of S.fusiforme seedlings.The excessive Cd significantly restricted photosynthesis and reduced photosynthetic pigments in the seedlings,resulting in growth inhibition and deep morphological color,especially of the stems.High levels of Cd in seawater had toxic effects on commercial S.fusiforme seedlings,and risked this edible seaweed for human food.展开更多
AIDS is a global pandemic that has seen the development of novel and effective treatments to improve the quality of life of those infected and reduction in the spread of the disease. While great advancements have been...AIDS is a global pandemic that has seen the development of novel and effective treatments to improve the quality of life of those infected and reduction in the spread of the disease. While great advancements have been made in HIV/AIDS therapeutics, there is still no cure or viable vaccine in development. The high rate of HIV-1 mutation contributing to virus immune escape, combined with increase in sexual transmission and the significant clinical therapeutics side effects of the currently available treatments highlights the need for novel therapeutics with broad anti-viral activity against both CXCR4 （X4） and CCR5 （R5）-tropic viruses. In our search for novel modalities against HIV infection, we investigated several aqueous extracts from Traditional Chinese Medicine （TCM） collection and identified Sargassum fusiforme （S. fusiforme） as a potent inhibitor of HIV infection. Following the Western approach of drug discovery and development, we isolated several bioactive molecules from S. fusiforme and determined their mechanism of action. TCM and Western approaches to disease treatment and drug development have been shown to be complementary to each other. The former provides a rich medicinal history for natural compounds that can have clinical impact on a variety of illnesses, while the latter enables the application of chemical informatics with rational drug design approach coupled to mechanistic underpinnings of such therapeutics. This multistep paradigm is demonstrated in the example of S. fusiforme as a treatment to prevent HIV infection, as described in this review.展开更多
Our previous study evaluated the in vitro and in vivo antioxidant activities of sulfated polysaccharides from a Celluclastassistedextract of Hizikia fusiforme (HFPS). The results indicate that HFPS possesses potent an...Our previous study evaluated the in vitro and in vivo antioxidant activities of sulfated polysaccharides from a Celluclastassistedextract of Hizikia fusiforme (HFPS). The results indicate that HFPS possesses potent antioxidant activity and suggestthe potential use of HFPS to combat photoaging. In this study, we investigated the ultraviolet (UV) protective effect of HFPSin vitro in keratinocytes (HaCaT cells) and in vivo in zebrafish. The results indicate that HFPS significantly reduced thelevel of intracellular reactive oxygen species (ROS) and improved the viability of UVB-irradiated HaCaT cells. In addition,HFPS remarkably decreased apoptosis formation in UVB-irradiated HaCaT cells in a dose-dependent manner. The in vivotest results also demonstrate that HFPS significantly reduced intracellular ROS levels, cell death, NO production, and lipidperoxidation levels in UVB-irradiated zebrafish in a dose-dependent manner. These results suggest that HFPS possessesstrong in vitro and in vivo UV-protective effects, making it a potential ingredient in the cosmeceutical industry.展开更多
BACKGROUND Various animal models have been used to explore the pathogenesis of choledochal cysts(CCs),but with little convincing results.Current surgical techniques can achieve satisfactory outcomes for treatment of C...BACKGROUND Various animal models have been used to explore the pathogenesis of choledochal cysts(CCs),but with little convincing results.Current surgical techniques can achieve satisfactory outcomes for treatment of CCs.Consequently,recent studies have focused more on clinical issues rather than basic research.Therefore,we need appropriate animal models to further basic research.AIM To establish an appropriate animal model that may contribute to the investigation of the pathogenesis of CCs.METHODS Eighty-four specific pathogen-free female Sprague-Dawley rats were randomly allocated to a surgical group,sham surgical group,or control group.A rat model of CC was established by partial ligation of the bile duct.The reliability of the model was confirmed by measurements of serum biochemical indices,morpho-logy of common bile ducts of the rats as well as molecular biology experiments in rat and human tissues.RESULTS Dilation classified as mild(diameter,≥1 mm to<3 mm),moderate(≥3 mm to<10 mm),and severe(≥10 mm)was observed in 17,17,and 2 rats in the surgical group,respectively,while no dilation was observed in the control and sham surgical groups.Serum levels of alanine aminotransferase,aspartate aminotrans-ferase,total bilirubin,direct bilirubin,and total bile acids were significantly elevated in the surgical group as compared to the control group 7 d after surgery,while direct bilirubin,total bilirubin,and gamma-glutamyltransferase were further increased 14 d after surgery.Most of the biochemical indices gradually decreased to normal ranges 28 d after surgery.The protein expression trend of signal transducer and activator of transcription 3 in rat model was consistent with the human CC tissues.CONCLUSION The model of partial ligation of the bile duct of juvenile rats could morphologically simulate the cystic or fusiform CC,which may contribute to investigating the pathogenesis of CC.展开更多
AIM:To describe the surgical procedure of fusiform penetrating keratoplasty(FPK)using multiple trephines of different sizes for treating patients with severe infectious keratitis.METHODS:Fourteen eyes underwent FPK,an...AIM:To describe the surgical procedure of fusiform penetrating keratoplasty(FPK)using multiple trephines of different sizes for treating patients with severe infectious keratitis.METHODS:Fourteen eyes underwent FPK,and 15 eyes received conventional penetrating keratoplasty(PK)were included in the study.The best-corrected visual acuity(BCVA),refractive outcomes,endothelial cell density,and postoperative complications were recorded.RESULTS:The FPK group was followed for an average of 15.3±2.1mo,whereas the PK group was followed for 16.1±1.9mo.The corneal ulcers were elliptical-shaped in all 14 eyes in the FPK group.The mean BCVA(logMAR,0.26±0.13)showed no statistically significant differences from that in the PK group(logMAR,0.21±0.12,P>0.05)at 1y after surgery.But the mean curvature,mean astigmatism,and mean spherical equivalent in the FPK group were lower than those in the PK group(P<0.05).Peripheral anterior synechia was observed in one patient in the FPK group,whereas 6 patients in the PK group.Suture loosening and neovascularization were observed in 4 and 5 eyes in the PK group,respectively.No graft immune rejection or elevation of intraocular pressure was observed in the two groups.CONCLUSION:For patients with elliptical-shaped corneas or corneal ulcers,FPK can avoid disrupting of corneal limbus,reduce the risk of postoperative complications,and can result in satisfactory visual quality.展开更多
基金supported by the grants from China Asia-Pacific Economic Cooperation(Nos.HV180101 and HV01-190101)the Xiamen Science and Technology Program(No.3502Z20161162)the Specific Fund Project of Xiamen Marine and Fishery Development(No.19CZP011HJ08).
文摘Tissue culture is one of the most promising and practical methods for conserving endangered plant species.Therefore,the present study evaluates the conservation of the endangered seaweed Sargassum fusiforme through tissue culturing techniques from different explants for the first time.Besides,the genetic variation of the mother plant and cultivated explants using inter-simple sequence repeat(ISSR)techniques;this methodology was the first to be illustrated in such work for algae tissue culturing.The regeneration results have shown that different explants could induce shoot and rhizoid morphogenesis with a total number of blades of 2436,1011,1466,678,and 6 from apical parts,stipe with blades,two-segmented seedlings,rhizoids,and stipe without blades,respectively.The total length was 234,181.8,83.5,and 81.8 cm from the two-segmented seedlings,apical parts,stipe without blades,and stipe with blades,respectively.At the same time,the total wet weight was 73.148,48.369,35.731,18.588,and 2.035 g from the apical parts,the two-segmented seedlings,stipe with blades,rhizoids,and stipe without blades,respectively.Micropropagation of S.fusiforme was successfully achieved with apical,stipe,and stolon segments using free PES media.It is suggested that the applied genetic fingerprint is valid for S.fusiforme and will respond well to molecular marker assistance in cultivation.The significance of S.fusiforme and its exposure to being endangered due to over-exploitation have made its regeneration in vitro a subject of interest in this study.Thus,this report represents the successful regeneration of S.fusiforme and explores the genetic uniformity or somaclonal variation of the obtained seedlings using the ISSR-PCR marker for the first time.
文摘[Objectives]This study was conducted to investigate the application of Sargassum fusiforme(Harv.)Setch.in cigarettes.[Methods]Tobacco-specific nitrosamines in the smoke of cigarettes added with ethanol extract of S.fusiforme were determined,and the compounds related to the aroma of S.fusiforme were identified by flavor-smelling experiment.[Results]With the addition of the ethanol extract of S.fusiforme,the decrease in the total amount of four tobacco-specific nitrosamines in mainstream cigarette smoke reached 16.42%.The results of the flavor-smelling experiment showed that the aroma of S.fusiforme might be related to(R)-5,6,7,7A-tetrahydro-4,4,7A-trimethyl-2(4H)-benzofuranone,glycerol,ethyl palmitate,methyl palmitate,ethyl linoleate,methyl(Z,Z,Z)-9,12,15-octadecatrienoate,ethyl(Z,Z,Z)-9,12,15-octadecatrienoate,phytol,and tetradecanoic acid.[Conclusions]The ethanol extract of S.fusiforme has the potential function of reducing the content of tobacco-specific nitrosamines in smoke and improving the taste of cigarettes.
基金Zhejiang Province Focuses on“Biological Engineering”Innovation Projects(No.CX2017001)the Autonomous Research Project of FSEKDNB(No.2020FSEKDNB001)。
文摘This study aimed to investigate the effects of Sargassum fusiforme polysaccharide(SFPS I,II,and III)on the apoptosis and regulation of human erythroleukemia(HEL)cells.The effect of different doses of SFPS on HEL cell growth was detected using the Cell Counting Kit-8 method,and apoptosis was detected by Hoechst staining.Cell cycle distribution and apoptosis were detected using flow cytometry.Expression of the cell cycle gene,p53,antiapoptotic genes,Bcl-xL and Bcl-2,and pro-apoptotic genes,Bax,Bad,and Caspase-3,as well as the expression of the corresponding proteins,were detected using real-time quantitative polymerase chain reaction(qPCR)and Western blot.The results showed that SFPS Ⅱ and Ⅲ decreased HEL cell viability and induced HEL cell apoptosis.Different concentrations of SFPS(Ⅰ,Ⅱ,and Ⅲ)were detected that induced much less toxic effect in normal human embryonic lung(MRC-5)cells,and SFPS Ⅰ increased cell proliferation,indicating its favorable selectivity towards cancer cells.The mechanism by which SFPS induced apoptosis was also found to be related to the induction of cell cycle arrest in the G0/G1 phase and the increased expression of apoptosis-related genes and proteins.We concluded that SFPS induces HEL cell apoptosis,possibly via activation of the Caspase pathway,providing the theoretical basis for the development of SFPS-based anti-tumor drug products.
基金Supported by the Ocean Public Welfare Scientifi c Research Project,State Oceanic Administration of the People’s Republic of China(No.201505033)the Qingdao People’s Livelihood Science and Technology Program(No.16-6-2-41-nsh)
文摘Heparin has been used as an anticoagulant drug for many years, but it has significant side ef fects. In the search for good substitutes, low molecular weight(MW) polysaccharides from S argassum fusiforme have been examined and confi rmed to possess biological activities. Here, S. fusiforme polysaccharides(SFP) were extracted and subjected to a hydrogen peroxide(H_2O_2) oxidation method for the preparation of low-MW SFP(LSFP). The effects of temperature, pH, and H_2O_2 concentration on the degradation process were also examined. Several LSFP of 36, 9, 5.7, and 2.7 kDa were obtained under different conditions, and their anticoagulant activities studied in vitro. The results showed that SFP and LSFP prolonged activated partial thromboplastin(APTT), prothrombin(PT) and thrombin times(TT) significantly, indicating that these low MW polysaccharides possessed anticoagulant activity in the intrinsic, extrinsic, and common coagulation pathways. As these ef fects were related to the MW of the polysaccharides in APTT and TT but not in PT, the contents of the monosaccharide fucose and sulfate and the polysaccharide MW could have exerted combined ef fects. The details of this mechanism require further verifi cation.
基金Supported by the Doutou Sci-Tech Project(No.N2006Y11B)the Shandong Key Sci-Technology Research Project(Nos.2018SDKJ0302-2,2018SDKJ0502-1)the CAS-Fujian STS Project(No.2017T3012)
文摘Since the 1980s,Sargassum fusiforme has been cultivated in Zhejiang,South China,and nowadays it becomes one of the important commercial seaweeds in China.With traditions of eating habits in the East Asian countries,this brown alga is used as food,because it contained functional oligo/polysaccharides and chemical components,and was regarded playing roles in antioxidant activities and regulating immunology.Through over 15 years’selection,breeding and cultivation,we obtained three strains with good traits and testified their characters during the production,which included the cultivars with high yield and other two good characters,either all the selected strains were applied in the Sargassum production.To avoid confusion during the selection and nursery,it was preferred to establish one fingerprint for distinguishing the Sargassum cultivars from different strains.Random amplified polymorphic DNA(RAPD)and inter-simple sequence repeat(ISSR)methods were adopted to analyze the genetic diversities of the selected S.fusiforme strains.With that,one fingerprint with RAPD markers was constructed,and one sequence characterized amplifi ed region(SCAR)marker to S.fusiforme was obtained.It is indicated that the applied fingerprint could be valid in S.fusiforme genetic and germplasm justification,and will be positive to molecular marker assistance in its selection and cultivation.
基金partially funded by Zhejiang Wanli University Scientific Research and Innivation Team(No.SC1032110880210)Zhejiang Provincial Top Discipline of Biological Engineering(No.KF2021010)Ningbo Public Service Platform for High-Value Utilization of Marine Biological Resources(Nos.NBHY-2017-S5,NBHY-2017(1))。
文摘Sargassum fusiforme(S.fusiforme)has been used as an ingredient in Chinese herbal medicine for thousands of years.However,there are a limited number of studies concerning its therapeutic mechanism.High performance gel permeation chromatography(HPGPC)analysis showed that the average molecular weight of the S.fusiforme polysaccharide,SFPS 191212,is 43 kDa.SFPS 191212 is composed of mannose,rhamnose,galactose,xylose,glucose,and fucose(at a molar ratio:2.1:2.9:1.8:15.5:4.6:62.5)withα-andβ-configurations.The present research evaluated the anti-tumor potential of the S.fusiforme polysaccharide in human erythroleukemia(HEL)cells in vitro.To explore the SFPS 191212’s apoptosis mechanism in HEL cells,transcriptome analysis was performed on HEL cells that were incubated with SFPS 191212.The inhibitory effect of SFPS 191212 on HEL cell growth was also analyzed.It was found that SFPS 191212 inhibited HEL cell proliferation,reduced cell viability in a concentration-dependent manner,and induced an insignificant toxic effect on normal human embryonic lung(MRC-5)cells.Compared with the control group,transcriptome analysis identified a total of 598 differentially expressed genes(DEGs),including 243 up-regulated genes and 355 downregulated genes.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed on all DEGs,and 900 GO terms and 52 pathways were found to be significantly enriched.Finally,23 DEGs were randomly selected and confirmed by quantitative real-time polymerase chain reaction(qRT-PCR).Moreover,SFPS 191212 down-regulated the PI3K/Akt signal transduction pathway.Our results provide a framework for understanding the effect of SFPS 191212 on cancer cells and can serve as a resource for delineating the anti-tumor mechanisms of S.fusiforme.
基金a part of the project titled‘Omics based on fishery disease control technology development and industrialization(20150242)’‘Development of functional food products with natural materials derived from marine resources(2017-0377)’funded by the Ministry of Oceans and Fisheries,Republic of Korea.
文摘Objective:To investigate whether ethanol extracts of Hizikia fusiforme could induce apoptosis in human prostate cancer PC3 cells.Methods:Cell viability was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.Apoptosis and mitochondrial membrane potential(MMP)were measured using flow cytometry in PC3 cells.DNA damage was assessed by nuclear staining and DNA fragmentation assay.Expressions of apoptosis-associated proteins were determined by Western blotting assays.Activities of caspase-3,-8,and-9 were determined by colorimetric assay.Moreover,intracellular reactive oxygen species(ROS)generation was detected using a flow cytometer and fluorescence microscope.Results:Treatment of PC3 cells with ethanol extracts of Hizikia fusiforme inhibited proliferation,which was associated with induction of apoptosis,and accompanied by increased expression of Fas,Fas-ligand(Fas L),Bax and t Bid,and decreased expression of Bcl-2.In addition,ethanol extracts of Hizikia fusiforme reduced c-Flip expression and activated caspase-8,-9 and-3,resulting in an increase in poly(ADP-ribose)polymerase(PARP)cleavage.However,in the presence of a pan-caspase inhibitor,ethanol extracts of Hizikia fusiforme-mediated growth inhibition and apoptosis were significantly attenuated.Ethanol extracts of Hizikia fusiforme also destroyed the integrity of mitochondria due to the loss of MMP,leading to cytosolic release of cytochrome c.Moreover,the levels of ROS were markedly increased by treatment with ethanol extracts of Hizikia fusiforme,which was significantly suppressed by the ROS scavenger N-acetyl-L-cysteine.Further investigation of whether ethanol extracts of Hizikia fusiforme-induced apoptosis was related to the generation of ROS was conducted and the results showed that N-acetyl-L-cysteine fully blocked ethanol extracts of Hizikia fusiforme-induced apoptotic events including loss of MMP,activation of caspase-3,the cytosolic release of cytochrome c and cytotoxicity.Conclusions:Ethanol extracts of Hizikia fusiforme have chemopreventive potential via induction of ROS-dependent apoptosis.Therefore,ethanol extracts of Hizikia fusiforme may be useful for developing effective and selective natural sources to inhibit cancer cell proliferation.
基金supported by the National High Technology Research and Development Program of China (863 Program) (2013AA093003)
文摘The response surface methodology(RSM) combined with bioassays was employed to optimize the extraction process of crude fucose-containing sulphated polysaccharides(c FCSP) from Sargassum fusiforme. The central composite design(CCD) was used with four variables, five levels, and four responses. The four variables were p H value of hydrochloric acid solution, extraction temperature(℃), ratio of liquid to raw material(m L g^(-1)), and extraction time(h), respectively. Chemical and bioassay indices were used in combination as the response parameters, which included the yield of c FCSP, fucose content, proliferation rate of spleen cells, and lipopolysaccharide-induced proliferation of splenocytes. The experimental data were fitted to a second-order polynomial equation using multiple regression analysis, and examined using the appropriate statistical methods. The best extraction conditions were as follows: the p H value of hydrochloric acid solution was 3.50; the extraction temperature was 100℃; the ratio of liquid to raw material was 15.00 m L g^(-1) and the extraction time was 2.50 h. The experimental yield was close to the predicted from the model. The extract could promote spleen lymphocyte proliferation, especially the lipopolysaccharide-induced lymphocyte proliferation in vitro, which suggested that its immunomodulatory effect on B lymphocytes. Therefore, c FCSP extracted from S. fusiforme could be utilized as an immunostimulant in functional foods and pharmaceutical industry in future.
文摘[Objectives] The research aimed to optimize extraction process of fucoidan from Sargassum fusiforme( FSF) and study its moisturizing performance. [Methods]Extracting condition of FSF by cellulase hydrolysis-ultrasonic assisted extraction method was optimized. The influences of solvent p H,enzyme dosage,extraction temperature,cellulose hydrolysis time,ultrasonication time,and the ratio of material to liquid on FSF were investigated by single factor and orthogonal experiments. [Results] The optimum extraction conditions were as followings:p H,4. 5; enzyme dosage,1%; extraction temperature,40℃; cellulose hydrolysis time,2 h; ultrasonic time,15 min; and the ratio of material to liquid,1∶ 10( g∶ m L). Under the optimal condition,the extraction yield of FSF was 8. 50%,RSD = 2. 74%. The short-time hygroscopicity( within 8 h) of crude extract of fucoidan from S. fusiforme( CEFSF) was better than glycerin,butanediol,and sodium alginate,and the moisture retention capacity of 1% CEFSF aqueous solution was better than 1% butanediol or 1% sodium alginate,and was equal to 5% glycerin under relative humidity of 43% and 81%. The determination results of skin moisture content and transepidermal water loss rate( TEWL)showed that: 5% CEFSF solution had good moisturizing effect. [Conclusions]The research could provide certain reference for deep development of S. fusiforme.
基金Foundation item:The National Key R&D Program of China under contract No.2018YFD0901500the National Natural Science Foundation of China under contract Nos 41706147 and 41876124.
文摘Cadmium(Cd)is a common heavy metal pollutant in the aquatic environment,generally toxic to plant growth and leading to growth inhibition and biomass reduction.To study the oxidation resistance in Sargassum fusiforme seedlings in response to inorganic Cd stress,we cultured the seedlings under two different Cd levels:natural seawater and high Cd stress.High Cd stress significantly inhibited the seedlings growth,and darkened the thalli color.Additionally,the pigment contents,growth rate,peroxidase(POD)activity,dehydroascorbic acid(DHA)content,and glutathione reductase(GR)activity in S.fusiforme were significantly reduced by high Cd treatment.Contrarily,the Cd accumulation,Cd2+absorption rate,dark respiration/net photosynthetic rate(Rd/Pn),ascorbic acid(Vc)content,soluble protein(SP)content,glutathione(GSH),and the activities of superoxide dismutase(SOD)and catalase(CAT)of S.fusiforme under Cd treatment significantly increased compared to the control group.The decrease of malondialdehyde(MDA)was not significant.Although S.fusiforme seedlings increased the antioxidant activities of POD,SOD,Vc,and the AsA-GSH cycle to disseminate H2O2 and maintain healthy metabolism,high Cd stress caused Cd accumulation in the stem and leaves of S.fusiforme seedlings.The excessive Cd significantly restricted photosynthesis and reduced photosynthetic pigments in the seedlings,resulting in growth inhibition and deep morphological color,especially of the stems.High levels of Cd in seawater had toxic effects on commercial S.fusiforme seedlings,and risked this edible seaweed for human food.
文摘AIDS is a global pandemic that has seen the development of novel and effective treatments to improve the quality of life of those infected and reduction in the spread of the disease. While great advancements have been made in HIV/AIDS therapeutics, there is still no cure or viable vaccine in development. The high rate of HIV-1 mutation contributing to virus immune escape, combined with increase in sexual transmission and the significant clinical therapeutics side effects of the currently available treatments highlights the need for novel therapeutics with broad anti-viral activity against both CXCR4 （X4） and CCR5 （R5）-tropic viruses. In our search for novel modalities against HIV infection, we investigated several aqueous extracts from Traditional Chinese Medicine （TCM） collection and identified Sargassum fusiforme （S. fusiforme） as a potent inhibitor of HIV infection. Following the Western approach of drug discovery and development, we isolated several bioactive molecules from S. fusiforme and determined their mechanism of action. TCM and Western approaches to disease treatment and drug development have been shown to be complementary to each other. The former provides a rich medicinal history for natural compounds that can have clinical impact on a variety of illnesses, while the latter enables the application of chemical informatics with rational drug design approach coupled to mechanistic underpinnings of such therapeutics. This multistep paradigm is demonstrated in the example of S. fusiforme as a treatment to prevent HIV infection, as described in this review.
文摘Our previous study evaluated the in vitro and in vivo antioxidant activities of sulfated polysaccharides from a Celluclastassistedextract of Hizikia fusiforme (HFPS). The results indicate that HFPS possesses potent antioxidant activity and suggestthe potential use of HFPS to combat photoaging. In this study, we investigated the ultraviolet (UV) protective effect of HFPSin vitro in keratinocytes (HaCaT cells) and in vivo in zebrafish. The results indicate that HFPS significantly reduced thelevel of intracellular reactive oxygen species (ROS) and improved the viability of UVB-irradiated HaCaT cells. In addition,HFPS remarkably decreased apoptosis formation in UVB-irradiated HaCaT cells in a dose-dependent manner. The in vivotest results also demonstrate that HFPS significantly reduced intracellular ROS levels, cell death, NO production, and lipidperoxidation levels in UVB-irradiated zebrafish in a dose-dependent manner. These results suggest that HFPS possessesstrong in vitro and in vivo UV-protective effects, making it a potential ingredient in the cosmeceutical industry.
基金the Key R&D Program of Zhejiang,No.2023C03029Health Science and Technology Plan of Zhejiang Province,No.2022RC201Zhejiang Provincial Natural Science Foundation Project,No.LY20H030007.
文摘BACKGROUND Various animal models have been used to explore the pathogenesis of choledochal cysts(CCs),but with little convincing results.Current surgical techniques can achieve satisfactory outcomes for treatment of CCs.Consequently,recent studies have focused more on clinical issues rather than basic research.Therefore,we need appropriate animal models to further basic research.AIM To establish an appropriate animal model that may contribute to the investigation of the pathogenesis of CCs.METHODS Eighty-four specific pathogen-free female Sprague-Dawley rats were randomly allocated to a surgical group,sham surgical group,or control group.A rat model of CC was established by partial ligation of the bile duct.The reliability of the model was confirmed by measurements of serum biochemical indices,morpho-logy of common bile ducts of the rats as well as molecular biology experiments in rat and human tissues.RESULTS Dilation classified as mild(diameter,≥1 mm to<3 mm),moderate(≥3 mm to<10 mm),and severe(≥10 mm)was observed in 17,17,and 2 rats in the surgical group,respectively,while no dilation was observed in the control and sham surgical groups.Serum levels of alanine aminotransferase,aspartate aminotrans-ferase,total bilirubin,direct bilirubin,and total bile acids were significantly elevated in the surgical group as compared to the control group 7 d after surgery,while direct bilirubin,total bilirubin,and gamma-glutamyltransferase were further increased 14 d after surgery.Most of the biochemical indices gradually decreased to normal ranges 28 d after surgery.The protein expression trend of signal transducer and activator of transcription 3 in rat model was consistent with the human CC tissues.CONCLUSION The model of partial ligation of the bile duct of juvenile rats could morphologically simulate the cystic or fusiform CC,which may contribute to investigating the pathogenesis of CC.
基金Supported by the National Natural Science Foundation of China (No.81900907,No.81870639)Taishan Scholar Program (No.20231255).
文摘AIM:To describe the surgical procedure of fusiform penetrating keratoplasty(FPK)using multiple trephines of different sizes for treating patients with severe infectious keratitis.METHODS:Fourteen eyes underwent FPK,and 15 eyes received conventional penetrating keratoplasty(PK)were included in the study.The best-corrected visual acuity(BCVA),refractive outcomes,endothelial cell density,and postoperative complications were recorded.RESULTS:The FPK group was followed for an average of 15.3±2.1mo,whereas the PK group was followed for 16.1±1.9mo.The corneal ulcers were elliptical-shaped in all 14 eyes in the FPK group.The mean BCVA(logMAR,0.26±0.13)showed no statistically significant differences from that in the PK group(logMAR,0.21±0.12,P>0.05)at 1y after surgery.But the mean curvature,mean astigmatism,and mean spherical equivalent in the FPK group were lower than those in the PK group(P<0.05).Peripheral anterior synechia was observed in one patient in the FPK group,whereas 6 patients in the PK group.Suture loosening and neovascularization were observed in 4 and 5 eyes in the PK group,respectively.No graft immune rejection or elevation of intraocular pressure was observed in the two groups.CONCLUSION:For patients with elliptical-shaped corneas or corneal ulcers,FPK can avoid disrupting of corneal limbus,reduce the risk of postoperative complications,and can result in satisfactory visual quality.
