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。

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。

Genetic analysis of selected Sargassum fusiforme (Harvey) Setchell (Sargassaceae, Phaeophyta) strains with RAPD and ISSR markers

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.

Ethanol extracts of Hizikia fusiforme induce apoptosis in human prostate cancer PC3 cells via modulating a ROS-dependent pathway

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.

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.

Preliminary exploration of animal models of congenital choledochal cysts

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.

Protective effect of sulfated polysaccharides from a Celluclast-assisted extract of Hizikia fusiforme against ultraviolet B-induced photoaging in vitro in human keratinocytes and in vivo in zebrafish

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.

羊栖菜不同分子质量褐藻多酚抗氧化活性研究

将羊栖菜(Sargassum fusiforme)经浸提、萃取和超滤等步骤获得不同分子质量的褐藻多酚。用普鲁士蓝法检测其总抗氧化能力;采用DPPH、羟自由基试剂盒、超氧阴离子试剂盒评价其对DPPH、羟自由基和超氧阴离子的清除能力。结果显示,羊栖菜褐藻多酚主要以高分子质量的形式存在;乙酸乙酯相组分各项抗氧化指标均好于水相,且乙酸乙酯相分子质量大于30 000组分对羟自由基与超氧阴离子的清除能力比没食子酸、Vc和VE强。

羊栖菜乙醇提取物的抗氧化活性

用体积分数70%的乙醇浸提羊栖菜(Sargassum fusiforme)得到粗提物,将粗提物溶于去离子水再依次用有机溶剂萃取,得到氯仿萃取物、乙酸乙酯萃取物、正丁醇萃取物和萃取后剩余物,测定粗提物和各萃取物的总多酚含量、维生素C和维生素E含量以及还原力。结果表明,正丁醇萃取物中的总多酚和维生素E含量较高,正丁醇萃取物和乙酸乙酯萃取物具有较强的还原力。

羊栖菜多糖抗肿瘤及其作用机制研究进展

羊栖菜(Sargassum fusiforme)多糖在多种肿瘤细胞系中表现出良好的抗癌活性,具有针对肿瘤细胞的选择活性和较小的毒副作用,可以作为现有肿瘤化疗药物的替代品进行开发。羊栖菜多糖主要通过诱导细胞凋亡作用于肿瘤细胞,其通过细胞周期停滞,增强机体免疫功能,改变细胞膜钙通道与流动性,破坏线粒体膜和产生一氧化氮来杀死癌细胞并防止转移。本文就国内外发表的羊栖菜多糖抗肿瘤功能及作用机制进行系统的归纳和总结,旨在为羊栖菜多糖抗肿瘤药物的深入研究、开发提供理论依据。

两种海藻提取物抗水产动物病原菌活性的初步研究

研究了两种大型海藻——羊栖菜(Sargassum fusiforme)、条浒苔(Enteromorpha clathrata)提取物对11种水产动物病原菌的抑制活性。结果显示:羊栖菜乙醇粗提物乙酸乙酯相对溶藻弧菌(Vibrio algi-nolyticus)的抑制作用最明显,抑菌圈可达21.6 mm;薄层层析表明羊栖菜乙醇粗提取物乙酸乙酯相有6条层析带,其中Rf=0.54带对哈氏弧菌(Vibrio harveyi)的抑制作用最强,抑菌圈可达13.6 mm;条浒苔乙醚粗脂对11种水产动物病原菌均表现出了较强的抑制作用,其中对灿烂弧菌(Vibrio splendidus)的抑制作用最强,抑菌圈可达24.7 mm;进一步的实验显示,条浒苔乙醚粗脂3种洗脱组分抗菌性均比粗脂减弱了,而其95%乙醇洗脱组分对恶臭假单胞菌(Pseudomonas putida)的抑菌圈仍可达13.8 mm。说明利用羊栖菜、条浒苔为开发海洋药物资源提供了可能。

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.

羊栖菜栽培敌害生物调查及其防治

本文主要通过洞头县羊栖菜人工栽培期间的生物调查,列举了羊栖菜栽培期间常见的敌害生物种类、分布和丰度情况。研究其危害性、危害途径、程度和防治技术,包括养殖户在多年的生产实践中已探索出卓有成效的防范措施和经验。希冀为洞头羊栖菜栽培业健康可持续发展有所贡献。

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.

Intravitreal aflibercept for ruptured retinal arterial macroaneurysm

Dear Editor,Retinal arterial macroaneurysm（RAM）,which usually occurs within the first three orders of arterial bifurcation,is acquired fusiform or saccular dilatations of retinal arterioles^（[1-2]）.Aging,the female gender,and systemic vascular