Application of Sargassum fusiforme(Harv.)Setch.Extract in Cigarettes

[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.

基于聚类及简化基因组联合分析的浙江洞头栽培羊栖菜(Sargassum fusiforme)品系筛选研究

以浙江洞头栽培羊栖菜(Sargassum fusiforme)群体为研究对象,运用主成分分析法对其鲜重、藻体长度、茎宽、气囊形态和大小等表型特征进行分析,确定了羊栖菜簇生气囊中的“特征”大气囊为表型主成分;运用聚类分析法分析了21个羊栖菜差异表型样品的“特征”大气囊表型聚类特征,定量分析归纳了栽培羊栖菜表型品系类别;运用简化基因组分析法构建了34个羊栖菜样品的系统进化树,分析了栽培羊栖菜各样品及韩国丽水、辽宁大连、浙江舟山野生羊栖菜样品间的亲缘关系,定量分析归纳了栽培羊栖菜基因型品系类别。在栽培羊栖菜表型品系和基因型品系总重合率为86%的基础上,将浙江洞头栽培羊栖菜群体分为球囊、锥囊、宽棒囊、狭棒囊和梭镖囊等5个品系,并运用林奈的“双名法”加以学名命名。提供了一种科学的羊栖菜品系分类方法,以期为深度开展浙江洞头栽培羊栖菜的群体结构、亲缘关系、遗传稳定性和品质差异等基础研究,以及定向选育优质高产、逆境高耐受性和活性物质高富集品系等应用研究,提供方法和理论支撑。

Preparation of low molecular weight Sargassum fusiforme polysaccharide and its anticoagulant activity

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.

栽培羊栖菜(Sargassum fusiforme)成熟藻体不同器官的组织结构显微观察及其生理生态学功能剖析

为全面了解栽培羊栖菜(Sargassum fusiforme)成熟藻体假根、茎、叶(气囊)和生殖托(雌托和雄托)四类器官的组织形态特征,运用石蜡切片法,对栽培羊栖菜成熟藻体各器官进行了组织形态学观察和讨论分析。结果表明,不同器官横切组织的形态、结构和宽度差异显著;假根,茎,叶,雌、雄生殖托的横切面由外向内分别为表皮、近表皮、皮层和髓部组织,而气囊无髓部组织结构,但具有内皮层。此外,雌、雄生殖托还包括生殖窝、卵和精子结构。栽培羊栖菜各器官表皮细胞均呈栅栏状排列,宽度顺序为:雄生殖托>茎>雌生殖托>气囊>叶>假根;表皮和近表皮细胞含淀粉粒最多,内含色素体;皮层和髓部细胞的淀粉粒含量相对较少;皮层组织的细胞腔最大,具有典型植物细胞形态和发达的胞间隙;髓部细胞致密,细胞腔多数较小。有关文献报道表明,羊栖菜不同器官响应光照、水温、干出与沉水及低盐胁迫等环境信号表达的生理生态学特征差异显著。研究结果可为深度解析栽培羊栖菜各器官之间的亚细胞结构差异,不同器官生理代谢响应环境信号表达差异,不同器官组织对微量元素或重金属离子的吸收、转运和储藏机制分析等研究提供基础数据和理论参考。

Bioassay-Guided Extraction of Crude Fucose-Containing Sulphated Polysaccharides from Sargassum fusiforme with Response Surface Methodology

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.

Extraction Process Optimization and Moisturizing Performance of Fucoidan from Sargassum fusiforme

[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.

Oxidative stress responses to cadmium in the seedlings of a commercial seaweed Sargassum fusiforme

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.

Effect of Sargassum fusiforme polysaccharide on apoptosis and its possible mechanism in human erythroleukemia cells

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.

Transcriptomic profile of human erythroleukemia cells in response to Sargassum fusiforme polysaccharide and its structure analysis

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.

Use of Sargassum fusiforme Extract and its Bioactive Molecules to inhibit HIV Infection: Bridging Two Paradigms between Eastern and Western Medicine

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.

Comparative analysis on transcriptome sequencings of six Sargassum species in China

Species of Sargassum are distributed worldwide, and are of great ecological and economic importance in marine ecosystems and bioresources. In this study, transcriptome sequencings of six Sargassum species were performed for the first time using an Illumina platform. For each sample, a total of 2.1-2.5 Gb of nucle-otides are collected and assembled into 69 871-116 790 scaffolds, with an average length of 410-550 bp and N50 length of 756-1 462 bp. A total of 20 512-28 684 unigenes of each sample were annotated and compared well with known gene sequences from nr database. Clusters of Orthologous Groups （COG）, gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） analyses were also performed for further un-derstanding of gene functions and regulation pathways. Gene expression levels were calculated based on RPKM values and compared among these species, especially for those genes related to carbohydrate metab-olism. Cluster analyses indicated that the differences of global gene expression between S. fusiforme, which was nominated as Hizikia fusiformis before, and other five species were not significant. Further phylogenet-ic analysis of 108 orthologous genes confirmed that S. fusiforme had closer relationship with S. hemiphyllum rather than S. horneri. These transcriptome data provided valuable information for better understanding of genome and gene characteristics of Sargassum algae and benefiting comparative and phylogenetic studies of Phaeophyceae species in future studies.

Adsorption of congo red on mesoporous activated carbon prepared by CO2 physical activation

This research demonstrates the production of mesoporous activated carbon from sargassum fusiforme via physical activation with carbon dioxide.Central composite design was applied to conduct the experiments at different levels by altering three operating parameters.Activation temperature(766-934℃),CO2 flow rate(0.8-2.8 L·min^-1)and activation time(5-55 min)were the variables examined in this study.The effect of parameters on the specific surface area,total pore volume and burn-out rate of activated carbon was studied,and the influential parameters of methylene blue adsorption value were identified employing analysis of variance.The optimum conditions for maximum methylene blue adsorption value were:activation temperature=900℃,activation time=29.05 min and CO2 flow rate=1.8 L·min(-1).The activated carbon produced under optimum conditions was characterized by BET,FTIR and SEM.The adsorption behavior on congo red was studied.The effect of parameters on the adsorbent dosage,temperature,PH and initial congo red concentration was investigated.The adsorption properties of the activated carbon were investigated by kinetics.The equilibrium removal rate and maximum adsorption capacity reaches up to 94.72%,234 mg·g^-1,respectively when initial congo red concentration is 200 mg·L^-1 under adsorbent dosage(0.8 g·L^-1),temperature(30℃),PH7.