Sulfated polysaccharides with antioxidant and anticoagulant activity from the sea cucumber Holothuria fuscogliva

Sea cucumber is a traditional nutritional food and medicinal resource with many bioactive components in China.Holothuria fuscogliva is a big sea cucumber with a rich of bioactive polysaccharides.To investigate the bioactivities of the polysaccharides from sea cucumber H.fuscogliva,we prepared the sulfated polysaccharides(HfP) from sea cucumber H.fuscogliva using a protease hydrolysis method.Antioxidant activities of HfP were investigated,including hydroxyl radical scavenging activity and superoxide radical scavenging activity.And,the anticoagulant activities of HfP were studied,including the activated partial thromboplastin time(APTT),prothrombin time(PT) and thrombin time(TT).The average molecular weight was 1 867.1 Da,with a sulfate content of 20.7%.In addition,the molar ratio of monosaccharide composition of HfP was Man:Rha:Glc A:Glc:Gal:Xyl:Fuc=0.083 6:0.437:0.134:0:1.182:0.748:1.It had a strong antioxidant activity,the hydroxyl and superoxide radical scavenging activity EC_(50) of HfP was 3.74 and 0.037 mg/mL,respectively.It also showed a good anticoagulant activity in our study.The APTT of HfP was much higher than that of heparin sodium,and the PT and TT of HfP was close to that of heparin sodium at a low concentration.Therefore,HfP shows a good antioxidant and anticoagulant activity and it may become a potential candidate of the natural antioxidant and anticoagulant and will have a good application future in health product or medicine industry.     

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.     

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.     

Molecular Weight Controllable Degradation of Laminaria japonica Polysaccharides and Its Antioxidant Properties

In this study, molecular weight controllable degradation of algal Laminaria japonica polysaccharides(LPS) was investigated by ultrasound combined with hydrogen peroxide. Three main factors, i.e., ultrasonic power(A), ultrasonic time(B), and H_2O_2 concentration(C) were chosen for optimizing parameters by employing three-factors, three-levels BBD. The influence of degradation on structure change and antioxidant activities was also investigated. A second-order polynomial equation including molecular weight(Y) of Laminaria japonica polysaccharides and each variable parameter, i.e., ultrasonic power(A), ultrasonic time(B), and H_2O_2 concentration(C), was established: Y=20718.67-4273.13A-4000.38B-1438.75C+2333.25AB+1511.00AC+873.00BC+2838.29A~2 + 2490.79B~2+873.04C~2. The equation regression coefficient value(R~2 = 0.969) indicated that this equation was valid. The value of the adjusted determination coefficient(adjusted R~2 = 0.914) also confirmed that the model was highly significant. The results of selected experimental degradation conditions matched with the predicted value. FT-IR spectra revealed that the structures of LPS before and after degradation were not significantly changed. Antioxidant activities of LPS revealed that low Mws possessed stronger inhibitory than the original polysaccharides. The scavenging effects on superoxide radicals was the highest when IC50 of crude LPS was 4.92 mg mL~(-1) and IC50 of Mw 18.576 KDa was 1.02 mg mL~(-1), which was fourfold higher than initial polysaccharide.     

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 amplified 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.     

Study on extraction of agaropectin from Gelidium amansii and its anticoagulant activity

Gelidium amansii agar was fractionated on DEAE-cellulose and four fractions were obtained sequentially. The yields of 1.0 mol/L NaCl fraction and 2.5 mol/L NaCl fraction were 2.80% and 2.03%. They are highly sulfated agar, and named as agaropectin with sulfate content being 22.8% and 32.5%, respectively. The anticoagulant experiment results show that agaropectin could effectively prolong the coagulation time in a dose-dependent manner in vitro. Agaropection could be absorbed and effectively prolong the plasma coagulation time in vivo. After intragastric administration at the doses of 100, 200, and 400 mg/kg-d in rats for 15 days, TT （thrombin time）, CT （coagulation time）, PT （prothrombin time）, and APTT （activated partial thromboplastin time） could be effectively prolonged and the plasma Fib level could be significantly lowered.     

The ecological risks of hydrogen peroxide as a cyanocide: its effect on the community structure of bacterioplankton

Microcystis blooms are an environmental and ecological concern that has received a serious attention. Hydrogen peroxide(H_2O_2) is an environment-friendly cyanocide that is commonly used to control Microcystis blooms. While the ecological safety of H_2O_2 has been previously studied, its influence on bacterioplankton has not been investigated to date. In this study, we used mesocosm experiments to determine the influence of H_2O_2 on the dynamic changes of the community structure of bacterioplankton. By using deep-sequencing and metagenomics strategy we determined the community structures of phytoplankton and bacterioplankton assemblages that were dominated by M icrocystis at a highly eutrophic Dianchi Lake, China. The results showed that M icrocystis was more sensitive to H_2O_2 than other eukaryotic algae. More interestingly, application of H_2O_2 changed the community structure of bacterioplankton, evidenced by the emergence of F irmicutes as the dominant species in place of B acteroidetes and Proteobacteria. The H_2O_2 treatment resulted in the community of bacterioplankton that was primarily dominated by E xiguobacterium and Planomicrobium. Our results show that the abundance changed and the bacterioplankton diversity did not recover even after the concentration of H_2O_2 reached to the background level. Thus, the response of bacterioplankton must be considered when assessing the ecological risks of using H_2O_2 to control Microcystis blooms, because bacterioplankton is the key player that forms the basis of food web of aquatic environment.     

PROMOTIVE EFFECT OF Se ON THE GROWTH AND ANTIOXIDATION OF A BLUE-GREEN ALGA SPIRULINA MAXIMA

ItwasreportedthatthegrOwthofthedrinediatomffelassforiraPsethenana(Hust.)HasleetHeiIndaldependedonSe(Priceetal.,1987).ThisreqUirementissPecificforSe;nootherelementscansubstituteforit,becauseSecaninduceglutathioneperoxidase(GSH-PoD)activityinthisdiatom(PriceandHarrison,l988),andinlow-CO2-grooCham~nasreinhandiiDangerad(Yokotaetal.,l988).ThisenZyInac-tivitywdetectedinextractsOfablue-greenalgaGleeoCaPasp.LB795(TotaandGallon,l979),andalsoincultUredhigherplancells(Drotaretal.,1985),butw…     

壳聚糖希夫碱铜配合物的制备及其催化性能

采用微波辐射加热法使壳聚糖与水杨醛反应生成壳聚糖希夫碱,然后将其与铜盐配位得到壳聚糖希夫碱铜配合物,用UV、FTIR、TG表征产物的结构,并探讨该配合物对过氧化氢分解的催化性能。研究结果表明:壳聚糖希夫碱铜配合物对过氧化氢有良好的催化活性,常温(25℃)常压,pH=7.0的条件下放置12 h,催化H2O2的分解达99.8%,该配合物重复使用3次还保持较好的催化活性。     

过氧化氢对全铁测定的影响

由过氧化钠分解试样的熔融物，经水提取后，会产生大量的过氧化氢。它对重铬酸钾测定全铁的结果，会产生偏高的影响。如果加热煮沸或以饿驱除它，则所得结果将令人满意。     

O_2和H_2O对脉冲电晕分解NO的影响

在脉冲电晕放电条件下,探讨了O2和H2O对NO分解反应过程的影响,并对复杂组分中NO的分解特性进行了实验研究。研究结果表明,脉冲电晕放电对NO分解反应具有一定的促进作用,可有效地分解模拟烟气中的NO,脱除效率随脉冲电压的增加而缓慢增加;受O2和H2O影响,NO分解转化率较N2氛围略有降低。     

Polyguluronate Sulfate,Polymannuronate Sulfate,and Their Oligosaccharides Have Antithrombin Ⅲ-and Heparin Cofactor Ⅱ-independent Anticoagulant Activity

Cardiovascular disease is the leading causes of death.However,the complications can be treated with heparin and heparinoids,such as heparin pentasaccharide Fondaparinux,dermatan sulfate,and PSS made from alginate extracted from brown seaweeds by chemical sulfation.Alginate is composed of a linear backbone of polymannuronate(PM),polyguluronate(PG),and alternate residues of mannuronic acid and guluronic acid.It is unknown if heparin and sulfated PG(PGS)/PM(PMS) have the same or different anticoagulant molecular targets.In the current study,the anticoagulant activities of PGS,PMS,and their oligosaccharides were directly compared to that of heparin,Fondaparinux,and dermatan sulfate by the activated partial thrombinplastin time(aP TT) assay using normal,antithrombin III(ATIII)-deficient,heparin co-factor II(HCII)-deficient,and ATIII-and HCII-double deficient human plasmas.Our results showed that PGS,PMS,and their oligosaccharides had better anticoagulant activity than that of Fondaparinux in all four human plasmas tested.As expected,heparin was the best anticoagulant in normal plasma.Moreover,PGS,PGS6,PGS12,PGS25,PMS6,PMS12,and PMS25 were better anticoagulants than dermatan sulfate in HCII-deficient plasma.Most strikingly,PGS,PGS12,PGS25,PMS6,PMS12,and PMS25 were better anticoagulants than that of heparin in ATIII-and HCII-double deficient human plasma.The results revealed for the first time that sulfated alginate had ATIII-and HCII-independent anticoagulant activities.Therefore,developing PGS and PMS-based anticoagulants might require to discover their major molecular targets and to develop target-specific anticoagulant assays.     

矿物包裹体的气、液相成分测定

矿物中包裹体温度、压力和气、液相成分的测定数据是阐明矿床成因、成矿物质来源、成矿物质化学条件和成矿元素运移方式等的重要地球化学参数。本文介绍了用载气热爆—气相色谱法测定气相成分(H_2、O_2、N_2、CH_4、CO、CO_2、H_2O)和用超声波提取液相成分,再用原子吸收分光光度计法和数字离子计法分别测定阳离子(K~+、Na~+、Ca~(2+)、Mg~(2+))、阴离子(F~-、Cl~-)。对实验装置、工作条件和分析步骤等本文均有叙述。并以石英标样,对实验结果作了初步讨论。其中对气相成分的提取,用载气热爆法代替真空热爆法可提高工作效率五倍。     

Polysaccharides in <Emphasis Type="Italic">Sipunculus nudus</Emphasis>: Extraction condition optimization and antioxidant activities

Marine organisms constitute unlimited resource of bioactive substances due to their high biodiversity and represent a valuable source of new compounds. This study optimized the alkali-extraction conditions and antioxidant activities of soluble polysaccharides from the body wall of Sipunculus nudus. The effects of solid-liquid ratio, extraction duration, extraction temperature, and alkali concentration on the yield of S. nudus polysaccharides (SNP) were examined, according to which the optimal combination of extraction parameters was obtained by an orthogonal test. The relative influencing importance of different extraction parameters on the yield of SNP followed the order as solid-liquid ratio > extraction temperature > alkali concentration > extraction duration. The highest extraction yield, 1.98%, was achieved under an optimal extraction condition: temperature 60°C, solid-liquid ratio 1:6 g mL^?1, duration 5 h, and alkali (NaOH) mass fraction 6%. The in vitro antioxidant activities examination showed that extracted SNP under this optimized condition had strong power in reducing certain hydroxyl and superoxide radical scavenging abilities. The promising results showed that extracted SNP could be a potent natural antioxidant.     

方斑东风螺两种养殖模式的比较

在室内水泥池,利用沙层自净养殖模式和直接铺沙养殖模式对不同规格的方斑东风螺(Babylonia areolataLink)进行了高密度养殖的研究。结果表明,沙层自净养殖模式养殖小螺、中螺、大螺组日均增重分别为0.031、0.088、0.098 g/d,沙层NH4+-N最高含量w分别为1.3、2.1、3.1 mg/L,H2S最高含量w分别为0.03、0.07、0.14mg/L,各规格组东风螺保持正常生长和活动,成活率92.9%以上;直接铺沙养殖模式养殖小螺、中螺、大螺日均增重分别为:0.023、0.051、0.068 g/d,成活率分别为95.2%、86.7%、84.9%,沙层NH4+-N最高含量w达到13.7mg/L,H2S最高含量w达到0.47 mg/L,沙层底质恶化,东风螺活动异常、不摄食。可见,沙层自净养殖模式对方斑东风螺的生长、成活率、沙层水质控制效果显著,在一定程度上克服了直接铺沙养殖底质恶化问题。     

类书的起源及其产生发展的原因探析

类书是一种特殊的文献编纂形式,它的真正起源应该是《皇览》。在文献聚集、文献利用、文学创作、帝王影响、文献编纂、学术思想等因素的共同作用下,类书得以产生与发展。类书是中国传统学术、文化与思想发展到魏晋南北朝时期的必然产物。     

Preparation and characterization of controlled-release fertilizers coated with marine polysaccharide derivatives

Encapsulation of water-soluble nitrogen fertilizers by membranes can be used to control the release of nutrients to maximize the fertilization ef fect and reduce environmental pollution.In this research,we formulated a new double-coated controlled-release fertilizer(CRF)by using food-grade microcrystalline wax(MW)and marine polysaccharide derivatives(calcium alginate and chitosan-glutaraldehyde copolymer).The pellets of water-soluble nitrogen fertilizer were coated with the marine polysaccharide derivatives and MW.A convenient and eco-friendly method was used to prepare the CRF.Scanning electron microscopy(SEM)and Fourier transform infrared spectroscopy(FTIR)were used to characterize the morphology and composition of the products.The nitrogen-release properties were determined in water using UV-Vis spectrophotometry.The controlled-release properties of the fertilizer were improved dramatically after coating with MW and the marine polysaccharide derivatives.The results show that the double-coated CRFs can release nitrogen in a controlled manner,have excellent controlled-release features,and meet the European Standard for CRFs.     

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.     

Spatio-temporal distribution of vegetation index and its influencing factors——a case study of the Jiaozhou Bay,China

The coastal zone is an area characterized by intense interaction between land and sea, high sensitivity to regional environmental changes, and concentrated human activities. Little research has investigated vegetation cover changes in coastal zones resulting from climate change and land-use change, with a lack of knowledge about the driving mechanism. Normalized diff erence vegetation index(NDVI) can be used as an indicator for change of the coastal environment. In this study, we analyzed the interannual changes and spatial distribution of NDVI in the coastal zone around Jiaozhou Bay in Qingdao, a coastal city undergoing rapid urbanization in northeast China. The underlying causes of NDVI variations were discussed in the context of climate change and land-use change. Results showed that the spatio-temporal distribution of NDVI displayed high spatial variability in the study area and showed a typical trend of gradually increasing from coastal to inland regions. The significant increase area of NDVI was mainly found in newly added construction land, extending along the coastline towards the inland. Land vegetation cover demonstrated a certain response relationship to sea-land climate change and land-based activities. The impact of land-based human activities was slightly greater than that of sea-land climate change for land vegetation cover. The results indicate that promoting ecological policies can build an ecological security framework of vegetation suitable for the resource characteristics of coastal cities. The framework will buf fer the negative ef fects of sea-land climate change and land-based human activities on vegetation cover and thereby achieve the balance of regional development and ecological benefits in the coastal zone.