Evaluation of antitumor, immunomodulatory and free radical scavenging effects of a new herbal prescription seaweed complex preparation

Seaweed Complex Preparation (SCP) is a clinical traditional Chinese medicine preparation which is composed of seven traditional Chinese herbs, and it has been used for treatment of lung cancer, liver cancer and digestive cancer. However, little information is available about the pharmacodynamic basis. The antitumor, immunomodulatory and free radical scavenging effects of SCP were evaluated in this study. Transplanted tumor in vivo method was used to determine the antitumor effect. The effects on splenocyte proliferation and phagocytosis of macrophages in tumor-bearing mice were measured by the MTT method and the phagocytizing cock red blood cell (CRBC) method respectively. The scavenging activities of SCP on DPPH and hydroxyl radicals in vitro were investigated. It was found that the medium-dose and high-dose of SCP could significantly inhibit the growth of transplanted hepatic tumor of murine hepatocarcinoma cell line H22, and promote proliferation of splenocytes and phagocytosis of macrophages. SCP possessed noticeable scavenging activities on DPPH and hydroxyl radicals. The antitumor effects of SCP might be achieved by improving immune system and scavenging free radicals, which is in accordance with the viewpoint of traditional Chinese medicine in promoting the body resistance and eliminating pathogenic factors for cancer treatment.     

Pond culture of seaweed Sargassum hemiphyllum in southern China

The seaweed Sargassum hemiphyllum is widely distributed throughout the coastal waters of Asia and has high commercial value.In recent years,its natural biomass has declined due to over-exploitation and environmental pollution.To seek for a feasible way to culture this seaweed efficiently,we designed a simple long-line system in a shrimp pond for the culture during winter,and the growth and nutritional composition of the seaweed were examined.Results show that the culture system was durable and flexible allowing S.hemiphyllum to grow vertically off the muddy bottom of the pond.Although the length of pondcultured S.hemiphyllum was inhibited by water depth,the weight-specific growth rate((1.65±0.17)%/d) was nearly three times higher than that of wild plants((0.62±0.19)%/d).The crude protein(6.92%±0.88%) and ash content(21.52%±0.07%) of the pond-cultured seaweed were significantly lower than those of the wild plants(9.38%±0.43% and 26.93%±0.07%,respectively);however,crude fat(1.01%±0.04%) was significantly higher than that of the wild plants(0.87%±0.02%).In addition,the nutritional composition of both pond-cultured and wild S.hemiphyllum was comparable to or even higher than those of other common seaweeds being used as food and/or aquaculture fodder.Future studies shall be focused on the impact of environmental parameters on its growth and nutritional composition.     

In-vitro anticoagulant activity of fucoidan derivatives from brown seaweed Laminaria japonica

Fucoidan, a group of sulfated heteropolysaccharides, was extracted from Laminaria japonica, an important economic alga species in China. The anticoagulant activity of fucoidan and its derivatives (including sulfated, phosphorylated, and aminated fucoidan) was examined using in-vitro anticoagulant systems. The correlation between chemical variations within the fucoidan group and anticoagulant activity was determined. The in-vitro anticoagulant properties of fucoidan and its derivatives were determined by measuring activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT). The results indicate anticoagulant activity in all samples using APTT and TT assays; however, only the fucoidan derivatives affected the PT assay. Thus, the fucoidan derivatives were able to inhibit both intrinsic and extrinsic blood coagulants. Fucoidan (FPS) and its derivatives presented better anticoagulant activity than low molecular weight fucoidan (DFPS) and its derivatives, suggesting that molecular weight and proper conformation are contributing factors for anticoagulant activity of polysaccharides. Amino groups have a positive charge and can thus change the charge density of fucoidan. Accordingly, among the tested samples, aminated fucoidan (NF) was the most active reflecting the importance of charge density for anticoagulant activity. Available data obtained using in-vitro models suggest that the sulfate content, sulfate/total-sugar ratio, molecular weight, and the substituted group of fucoidan are important factors for anticoagulant activity but that the influence of sulfate, phosphate and amino groups on anticoagulant activity was different.     

IN VITRO STUDY ON CYTOTOXIC EFFECTS OF THE ORGANOPHOSPHOROUS PESTICIDE PROFENOFOS ON THE GILL CELL LINE,FG-9307, OF THE FLOUNDER （PARALICHIHYS OLIVACEUS）

The FG-9307 cell line derived from the gill of flounder Paralichthys olivaceus was used in the present study to determine the acute cytotoxic effects of the organophosphorous pesticide, profenofos. It was found that the cell growth rate was markedly reduced by profenofos at the concentrations of 2.5, 5 and 7.5 mg/L tested; and that the fine structures of the cells were also altered by profenofos, as evidenced by dilation of nuclear membranes and mitochondria cristae, and presence of enlarged lysoscrneswith engulfed organelles and numerous vacuoles in the cytoplama. Probably, mitochondria, the cell ener-gy-generating sites, are the most prcrninent sites of profenofos cytotoxity in the cells. This seems to be the first report of the use of marine fish cell line for evaluation of the acute in vitro cytotoxicity of organophos-phorus pesticide.     

In vitro study of neuroendocrine regulation over the testicular development in mud crabs Scylla serrata

1 INTRODUCTION Ovarian development of female crustaceans is under the direct regulations of two neurohormones: gonad-inhibiting hormone (GIH) secreted by optic ganglia and gonad-stimulating hormone (GSH) se- creted by brain and thoracic ganglia (Fingerman…     

In vitro acute cytotoxicity of abamectin to the Gill Cell Line of Flounder Paralichthy olivaceus

The cytotoxicity of abamectin to the Gill Cell Line of Flounder (FG cell line) was examined in this study. It was found that the exposure of FG cells to abamectin caused the decreases of both cell growth rate and antioxidant enzyme activities,and the increase of intracellular O2- content. It was proposed that the reduction of antioxidant enzyme activities in FG cells caused the accu- mulation of O2- content in FG cells, leading to the change of cell morphology and even the death of cells. The results showed that FG cell line is suitable for the evaluation of the acute toxicity of abamectin.     

Extracts of the seaweed Bifurcaria bifurcata display antifungal activity against human dermatophyte fungi

Bifurcaria bifurcata is a seaweed of the Order Fucales(Ochrophyta, Phaeophyceae) that can be found all year round along the Portuguese Atlantic shore. Although it is considered edible in some countries,its biotechnological potential has not yet been assessed in detail and here we report its antimicrobial potential against human dermatophyte fungi. Three samples were harvested from Baleal Island(Peniche, Portugal)harvested in winter(BbPe); from Aguda Beach(Arcozelo, Vila Nova de Gaia, Portugal), harvested in autumn(BbAg) and a sample obtained by culturing the Aguda sample under laboratory conditions(BbLC).A broth macrodilution assay was applied to determine the MIC(minimum inhibitory concentration) and MLC(minimum lethal concentration) of the B. bifurcata extracts. Results show that the BbPe MeOH extracts were the most effective and had higher antifungal activity against all the tested dermatophyte strains than the BbLC and BaAg extracts [Epidermophyton floccosum FF9(100-200 μg/mL MIC; 200 μg/mL MLC), Microsporum canis FF1(400 μg/mL MIC; 400 μg/mL MLC), Trichophyton mentagrophytes FF7(100 μg/mL MIC; 800 μg/mL MLC), M. gypseum CECT 2908(800 μg/mL MIC; 800 μg/mL MLC), T.mentagrophytes var. interdigitale CECT 2958(800 μg/mL MIC; 800 μg/mL MLC), T. rubrum CECT 2794(200 μg/mL MIC; 400 μg/mL MLC)]. In fact, only E. floccosum FF9 was sensitive to BbAg and BbLC MeOH extracts. To our knowledge, this is the first report of antifungal activity of B. bifurcata against human dermatophyte fungi.     

COMPOSITION AND PROPERTIES OF AGAR-TYPE POLYSACCHARIDES FROM GRACILARIA SJOESTEDTII KYLIN

A study was made on the composition and properties of agar-type polysaccharides of Gracilaria sjoestedtii from Qingdao, China. Fractional extraction was carried out and four fractions were obtained. The yields and chemical composition of these fractions were determined. Gel strength measurements of the agar-type polysaccharide fractions were made after alkli modification. The nature of the non-gelling, highly sulphated galactans (Fraction II) present in the thaw water after the initial extraction of Gracilaria in a freeze-thaw cycle was reported and discussed.     

波纹龙虾胚胎的离体培养及发育观察

对波纹龙虾不同发育时期的胚胎进行离体培养和显微观察。结果表明,初始培育的离体胚胎愈接近孵化期,其孵化率愈高(最高达80.0%);波纹龙虾胚胎发育经历受精卵、卵裂期、囊胚期、原肠期、中眼色素形成期、复眼色素形成期、心跳期、破膜前期和出膜期;胚胎在发育过程中颜色变化:橙红→深橙红→砖红→灰白→接近透明。     

Development of a seaweed species-selection index for successful culture in a seaweed-based integrated aquaculture system

Integrated multi-trophic aquaculture (IMTA) has been proposed as a concept that combines the cultivation of fed aquaculture species (e.g., finfish/shrimp) with extractive aquaculture species (e.g., shellfish/seaweed). In seaweed-based integrated aquaculture, seaweeds have the capacity to reduce the environmental impact of nitrogen-rich effluents on coastal ecosystems. Thus, selection of optimal species for such aquaculture is of great importance. The present study aimed to develop a seaweed species-selection index for selecting suitable species in seaweed-based integrated aquaculture system. The index was synthesized using available literature-based information, reference data, and physiological seaweed experiments to identify and prioritize the desired species. Undaria pinnatifida, Porphyra yezoensis and Ulva compressa scored the highest according to a seaweed-based integrated aquaculture suitability index (SASI). Seaweed species with the highest scores were adjudged to fit the integrated aquaculture systems. Despite the application of this model limited by local aquaculture environment, it is considered to be a useful tool for selecting seaweed species in IMTA.     

Harmful algal bloom-forming dinoflagellate Prorocentrum donghaiense inhibits the growth and photosynthesis of seaweed Sargassum fusiformis embryos

Journal of Oceanology and Limnology - Harmful algal bloom (HAB) is an ecological disaster to local mariculture. At present, its impact on macrophytes has not been well studied. In this study, we...     

In vitro Antioxidant Effects of Porphyra haitanensis Peptides on H_2O_2-Induced Damage in HepG2 Cells

In this study,protein from Porphyra haitanensis was used as raw material to prepare an antioxidant peptide,and its antioxidant activity was evaluated in vitro.A model of H_2O_2-induced oxidative damage in Hep G2 cells was established,and the effects of Porphyra haitanensis hydrolysates (PHHs) on superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were detected.Finally,the structure of PHHs was identified by ESI-MS/MS.The results showed that the 1,1-diphenyl-2-pyridylhydrazine(DPPH)-free radical-scavenging ability of PHHs was the strongest (59.28%at 1.0 mg m L~(-1)) when hydrolyzed with an acidic protease for 4 h.PHHs with different concentrations had protective effects on H_2O_2-induced damage to Hep G2 cells,and the protective effect was enhanced with increasing concentrations.When the level was 400μg m L~(-1),the cell survival rate was as high as 88.62%.Moreover,PHHs can significantly reduce oxidative damage to Hep G2 cells by H_2O_2,improve SOD activity,and reduce MDA content.The tetrapeptide Asp-Lys-Ser-Thr,with a molecular weight of 448 Da,was identified as an important fraction of PHHs by high-resolution mass spectrometry.     

飞机草总黄酮体外抑菌活性与体内抗大肠埃希菌感染效果

提取飞机草(Eupatorium odoratum Linn.)总黄酮,从抑菌圈、最小抑菌浓度(MIC)及最小杀菌浓度(MBC)等方面探讨飞机草总黄酮对金黄色葡萄球菌(Staphylococcus aureus)、鸡白痢沙门氏菌(Salmonella pullorum)、枯草芽孢杆菌(Bacillus subtilis)、大肠埃希菌(Escherichia coli)的体外抑菌活性;经腹腔注射大肠埃希菌感染小鼠后,灌胃给予飞机草总黄酮,观察供试小鼠腹泻率,测定小鼠粪便大肠埃希菌数及血清抗体水平(IgM、IgA和IgG),评价飞机草总黄酮的体内抗感染作用。结果表明,金黄色葡萄球菌对飞机草总黄酮最为敏感,其抑菌圈直径为19.50 mm,MIC为0.015 6 g/m L,MBC为0.031 3 g/m L;160 mg/kg的飞机草总黄酮可降低小鼠腹泻率和粪便大肠埃希菌数(P0.01),提高血清IgA(P0.01)和IgG含量(P0.05)。飞机草总黄酮具有体内、体外抗菌作用。     

A comparative analysis of four kinds of polysaccharides purified from Furcellaria lumbricalis

Four kinds of polysaccharides (FA, FB, FC and FD) were extracted and purified from a red alga Furcellaria lumbricalis. Their monosaccharide composition, average molecular mass and structural feature were determined by gas chromatography (GC), high performance gel permeation chromatography (HPGPC) and Fourier transform infrared spectroscopy (FTIR), respectively. The results showed that FA and FB were composed of galactose (Gal), 3,6-anhydro-galactose (AnG) and 4-sulfated-galactose (G4S), which were structurally similar to β-carrageenan and κ-carrageenan. FC was composed of sulfated galactose and small amount of AnG, indicating that it was a hybrid-sulfated galactan. FD was composed of α-1,4-glucose and belonged to floridean starch. The respective yields of FA, FB, FC and FD were 4.84%, 33.74%, 8.76% and 13.5%, and the corresponding were 537 kD, 432 kD, 513 kD and 51 kD respectively. Owning to its high content (>60%) of polysaccharide, F. lumbricalis will be a good source of marine sulfated polysaccharide and shows its potential utilization in food and medicine.     

Preparation of polysaccharides in different molecular weights from Ulva pertusa Kjellm （Chorophyta）

As molecular weight controls the biological activities of polysaccharides, screening the optimal molecular weight of polysaccharides is important in drug research and application. In this study, hydrogen peroxide was employed as oxidant, and temperature, reaction time, and concentration of polysaccharides and hydrogen peroxide were examined for their effects on the preparation of polysaccharides in different molecular weights from Ulva pertusa. Our experiment suggested that the optimal degradation concentrations for polysaccharides and hydrogen peroxide were 2.5% (w/v) and 8.0% (v/v), respectively. The range of degradation measured by relative viscosity was mainly controlled by temperature and time. Results revealed that 35℃ was the optimal temperature for obtaining low-degradation samples, and 50℃ was the most favorable temperature to accelerate the reaction to yield highly-degraded samples. Four samples in different molecular weights A, B, C and D were finally prepared. The controllability was evaluated by the relative standard deviation (RSD) of relative viscosity, and the peak molecular weights and the polydispersity indexes (Mw/Mn) of molecular weights were measured by high performance gel permeation chromatography (HPGPC).     

Structural analysis and anti-complement activity of polysaccharides extracted from Grateloupia livida (Harv.) Yamada

Polysaccharides were extracted from Grateloupia livida(Harv.) Yamada using hot water(extracted product denoted WGW) and then degraded in dilute sulfuric acid(degraded product denoted WGWD). The degraded mixture was then separated into four fractions through anion exchange chromatography on 2-diethylaminoethanol(DEAE)-Bio-Gel Agarose FF gel. Electrospray ionization collision-induced dissociation tandem mass spectrometry(ESI-CID-MS/MS) was performed to elucidate the structural features of all fractions. In combination with nuclear magnetic resonance spectroscopy(NMR)and infrared spectroscopy(IR) data, the major polysaccharide structures were concluded to be μ-carrageenan and κ-carrageenan. μ-Carrageenan usually has a backbone of alternating 1,3-linked β-D-galactopyranose residues sulfated at C-4 and 1,4-linked a-D-galactopyranose residues sulfated at C-6, while κ-carrageenan consists of alternating 1,3-linked β-D-galactopyranose residues sulfated at C-4 and 1,4-linked a-D-3,6-anhydrogalactopyranose residues. Trace v-carrageenan, composed of 1,3-linked β-D-galactopyranose residues sulfated at C-4 and 1,4-linked a-D-galactopyranose residues sulfated at C-2 and C-6, was also detected. Furthermore, the polysaccharide had a backbone comprising 1,3-linked β-D-galactopyranose and1,4-linked α-L-galactopyranose sulfated at C-6, which is the agarose precursor. The hydroxy groups in the galactopyranose were partially substituted by methyl and pyruvic acid acetal(PA) groups. The anticomplementary activities of WGW and its derivatives against classical pathways were measured. The native polysaccharides in WGW had higher activities, while the derivatives had much weaker activities. This indicated that the molecular weight and sulfate content were important factors affecting the anti-complement activity.     

Properties of polysaccharides in several seaweeds from Atlantic Canada and their potential anti-influenza viral activities

To explore the polysaccharides from selected seaweeds of Atlantic Canada and to evaluate their potential anti-influenza virus activities, polysaccharides were isolated from several Atlantic Canadian seaweeds, including three red algae (Polysiphonia lanosa, Furcellaria lumbricalis, and Palmaria palmata), two brown algae (Ascophyllum nodosum and Fucus vesiculosus), and one green alga (Ulva lactuca) by sequential extraction with cold water, hot water, and alkali solutions. These polysaccharides were ana-lyzed for monosaccharide composition and other general chemical properties, and they were evaluated for anti-influenza virus activities. Total sugar contents in these polysaccharides ranged from 15.4% (in U. lactuca) to 91.4% (in F. lumbricalis); sulfation level was as high as 17.6% in a polysaccharide from U. lactuca, whereas it could not be detected in an alikali-extract from P. palmaria. For polysaccharides from red seaweeds, the main sugar units were sulfated galactans (agar or carrageenan) for P. lanosa, F. lumbricalis, and xylans for P. palmata. In brown seaweeds, the polysaccharides largely contained sulfated fucans, whereas the polysaccharides in green seaweed were mainly composed of heteroglycuronans. Screening for antiviral activity against influenza A/PR/8/34 (H1N1) virus revealed that brown algal polysaccharides were particularly effective. Seaweeds from Atlantic Canada are a good source of marine polysaccharides with potential antiviral properties.     

Chemical characteristics and anticoagulant activities of two sulfated polysaccharides from Enteromorpha linza (Chlorophyta)

Two sulfated polysaccharides, designated MP and SP, were extracted from the marine green alga Enteromorpha linza using hot water and then purified using ion-exchange and size-exclusion chromatography. The anticoagulant activities of MP and SP were examined by determination of their activated partial thromboplastin time (APTT), thrombin time (TT) and prothrombin time (PT) using human plasma. Results showed that MP and SP were composed of abundant rhamnose with small amounts of xylose and glucuronic acid, whereas SP also contained a small amount of galactose. Approximate molecular weights of MP and SP were 535 and 502 kDa, respectively. As compared with SP, MP had higher contents of sulfate ester (19.0%) and uronic acid (14.9%). The MP mainly consisted of (1→4)-linked rhamnose residues with partially sulfated groups at the C-3 position, and small amounts of (1→3, 4)-linked rhamnose, (1→2, 4)-linked rhamnose, (1→4)-linked glucuronic acid and (1→4)-linked xylose residues. The SP contained abundant (1→4)-linked rhamnose with minor amounts of (1→3)-linked rhamnose, (1→3, 4)-linked rhamnose, (1→2, 4)-linked rhamnose, (1→4)-linked glucuronic acid, (1→4)-linked xylose, and (1→3)-linked galactose residues. The sulfate groups were mainly located at C-3 of (1→4)-linked rhamnose residues. Both MP and SP, in particular the former, effectively prolonged APTT and TT. This work demonstrates that MP and SP have unique structural characteristics distinct from those of other sulfated polysaccharides from Enteromorpha. The MP is a potential source of anticoagulant, and the difference in anticoagulant activities of the two sulfated polysaccharides is directly linked to the discrepancy of their chemical features.     

Comparative studies on structural feature of agar polysaccharides fromPorphyra haitanensis grown in south and north China

The structural feature of agar polysaccharides fromPorphyra haitanensis grown in south China and transplanted to the north was investigated by fractionation on DEAE—Sephadex A 50, chemical analysis, and infrared and¹³C-NMR spectroscopy. The agars composed mainly of charged molecules were eluted from DEAE—Sephadex A 50 with 1.0 mol/L NaCl solution from the southernP. haitanensis and with 0.5 mol/L NaCl from the northern one. The¹³C-NMR spectra showed that agarobiose [(1→3)-β-D-galactopyranosyl-(1→4)-3,6-anhydro-α-L-galactopyranose] and the biological precursor of agarobiose [(1→3)-β-D-galactopyra nosyl-(1→4)-6-sulfate-α-L-galactopyranose] were the major disaccharide repeating units in the charged fractions. The content of the biological precursor in the agar polysaccharides from southernP. haitanensis was higher than that in the northern one, the content of the biological precursor extracted from cold water was higher than that from hot water, and the content of 6-OMe-D-galactose in the southernP. haitanensis polysaccharides was higher than in the northern one. This distinct difference will be of significance for further study of the physiological characters ofP. haitanensis. Contribution No. 1849 from the Institute of Oceanology, Academia Sinica.     

Preparation of polysaccharides in different molecular weights from Ulva pertusa Kjellm (Chorophyta)

As molecular weight controls the biological activities of polysaccharides, screening the optimal molecular weight of polysaccharides is important in drug research and application. In this study, hydrogen peroxide was employed as oxidant, and temperature, reaction time, and concentration of polysaccharides and hydrogen peroxide were examined for their effects on the preparation of polysaccharides in different molecular weights fromUlva pertusa. Our experiment suggested that the optimal degradation concentrations for polysaccharides and hydrogen peroxide were 2.5% (w/v) and 8.0% (v/v), respectively. The range of degradation measured by relative viscosity was mainly controlled by temperature and time. Results revealed that 35°C was the optimal temperature for obtaining low-degradation samples, and 50°C was the most favorable temperature to accelerate the reaction to yield highly-degraded samples. Four samples in different molecular weights A, B, C and D were finally prepared. The controllability was evaluated by the relative standard deviation (RSD) of relative viscosity, and the peak molecular weights and the polydispersity indexes (M_w/M_n) of molecular weights were measured by high performance gel permeation chromatography (HPGPC). This work was supported by the Scientific and Technical Bureau of Shandong Province.