Effects of carboxymethyl-chitosan on wound healing in vivo and in vitro

Effects of carboxymethyl-chitosan with different molecular weights on wound healing were investigated in vivo and in vitro.A second degree burn model was performed in rats and the accelerative effects of carboxymethyl-chitosan on wound repair were observed.Contents of transforming growth factor(TGF)-β1,interleukin(IL)-6 and matrix metalloproteinase(MMP)-1 in wounds were determined by enzyme-linked immunosorbent assay(ELISA).In vitro study evaluated the influence of carboxy-methyl-chitosan on cytokines secretion of fibroblasts and macrophages.In vivo results showed that carboxymethyl-chitosan effec-tively accelerated the wound healing process in burned rats(P<0.05).Levels of TGF-β1,IL-6 and MMP-1 in carboxymethyl-chitosan groups were significantly elevated,compared with control group(P<0.05).In vitro results indicated that carboxymethyl-chitosan significantly promoted the proliferation of fibroblasts and stimulated its IL-8 and IL-10 secretion at different incubation time,but it did not affect collagen secretion of fibroblasts.Carboxymethyl-chitosan enhanced phagocytosis ability of macrophages,and in-creased its tumor necrosis factor(TNF)-α secretion.In conclusion,carboxymethyl-chitosan promoted wound healing by activating macrophages,accelerating fibroblasts growth and exerting considerable effects on the secretion of a series of cytokines.     

Effect of fossil diatoms on the growth of two syngeneic murine leukemias

The effect of micronized frustules of fossil diatoms (DS) on tumor growth and survival of mice receiving an ip inoculations of either a spontaneous lymphoblastic leukemia (LB) or P-388 lymphocytic leukemia (P-388) was investigated. A multi-dose study was designed, mice receiving 1000, 100, 10 and 1 μg of a suspension of DS, either ip or sc on day 1, 4 and 9 after tumor inoculation. With LB the best results were observed when DS were inoculated by the ip route, no effect being observed with the sc route. The best therapeutic effect resulted with 1 μg DS: 33% of the mice were alive 24 days after tumor inoculation, when all the controls had died. With P-388, only a temporary effect with the 10 μg DS dose was observed. In no case was it possible to prolong survival. It is concluded that at low doses of DS therapeutic effect is greatest, while toxicity due to SiO₂ is greatly reduced.     

A study on the meat and bone meal or poultry by-product meal as protein substitutes of fishmeal in concentrated diets forParalichthys olivaceus

A study was conducted to evaluate the effects of meat and bone meal （MBM） and poultry by-product meal （PBM） as the replacement of fishmeal in the diets on the growth performance, survival and apparent digestibility coefficient （ADC） of Japanese flounder （Paralichthys olivaceus）. The experimental diets included 0%, 20%, 40%, 60% and 80% MBM or PBM replacement of total fishmeal respectively. All diets were iso-nitrogenous and isocaloric. The results showed that there are no significant differences （P 〉0.05） in growth performance among the treatments fed with 0% -60% MBM replacement of fishmeal, while the percent weight gain （WG, % ）, body length gain （BLG, % ） and ADC significantly decrease when fishmeal is replaced by 80% MBM. The result showed also that there are no significant differences （P 〉0.05） in growth performance and ADC among all treatments fed with the diets with 0% -80% replacements of fishmeal with PBM.     

A Study on the Meat and Bone Meal and Poultry By-product Meal as Protein Substitutes of Fish Meal in Practical Diets for Litopenaeus vannamei Juveniles

1　Introduction　Proteinisanimportantnutritionalingredientofaquaticfeed .Thehighproteinneedsoffish ,shrimpandmostotheraquaticanimalsissuppliedmainlybyfishmeal (WilliamsandBarlow ,1996 ) .Recently ,thefishmeal,asamainproteinsourceofaquaticfeedisshort suppliedintheinternationalmarketanditspriceisbecominghigherandhigher .Therefore ,itisnecessarytofindsubstitutesoffishmealasaproteinsourceofaquaticfeed .　Someotheranimalby productmealssuchasmeatandbonemeal (MBM )andpoultryby productmeal(PBM )ha…     

A study on the meat and bone meal and poultry by-product meal as protein substitutes of fish meal in practical diets for Litopenaeus vannamei juveniles

A study was conducted to evaluate the effects of meat and bone meal (MBM) and poultry by-product meal (PBM) as the replacement of fish meal in the diets on the growth performance, survival and apparent digestibility coefficients (ADC) of Litopenaeus vannamei. The basal diets were formulated with 22% fish meal and other ingredients which provided about 40% protein and 9% lipid in the diet. The experimental diets included MBM or PBM to replace 0, 20%, 40%, 60% and 80% of total fish meal respectively. All diets were iso-nitrogenous and isocaloric in gross terms. The results showed that there were no significant differences (Pτ;0.05) in growth performance and ADC among the treatments fed with the diets in which 0–60% fish meal had been replaced with MBM, while the percent weight gain (WG, %), body length gain (BLG, %) and ADC significantly decreased when the MBM was up to 80% of the fish meal. There were no significant differences (Pτ;0.05) in growth performance and ADC among all the treatments fed with the diets in which 0–80% fish meal had been replaced with PBM.     

The polypeptide in Chlamys farreri can protect human dermal fibroblasts from ultraviolet B damage

To investigate the effect of polypeptide fromChlamys farreri (PCF) on NHDFin vitro, we modeled oxidative damage on normal human dermal fibroblasts (NHDF) exposed to ultraviolet B (UVB). In this study, 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) were tested to measure cell viability. Enzymes including superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), catalase (CAT) and xanthine oxidase (XOD) were determined biochemically. Total antioxidative capacity (T-AOC) and anti-superoxide anion capacity (A-SAC) were also determined. Ultrastructure of fibroblasts was observed under transmission electron microscope. The results showed that: UVB (1.176×10⁻⁴ J/cm²) suppressed the growth of fibroblasts and the introduction of PCF (0.25%–1%) before UVB reduced the suppression in a concentration-dependent manner. PCF could enhance the activities of SOD, GSH-PX and T-AOC as well as A-SAC. Also PCF could inhibit XOD activity, while it did not affect CAT activity. Ultrastructure of fibroblasts were damaged after UVB irradiation, concentration-dependent PCF reduced the destructive effect of UVB on cells. These results indicated that PCF can protect human dermal fibroblasts from being harmed by UVB irradiation via its antioxidant proerty. This work was supported by the National Natural Science Foundation of China (No. 39970638) and the Science and Technology Bureau of Qingdao (No. 2001-28-50).     

复方中草药对吉富罗非鱼生长及肠道菌群的影响

将A、B、C三种复方中草药分别按质量分数1.5%的比例添加到罗非鱼商品饲料中,喂养初始体重约16.58±0.48g的吉富罗非罗28d,通过测定罗非鱼的增重率、饲料系数、肠道菌群等指标,研究三种复方中草药制剂对罗非鱼生长性能及肠道菌群的影响。结果显示:三种复方中草药制剂对罗非鱼的增重率、成活率、饲料系数等生长性能指标虽无显著影响(P>0.05),但与对照组相比,添加1.5%的复方中草药制剂C可使吉富罗非鱼的增重率提高10.82%,饲料系数降低4.71%,蛋白质效率提高15.68%;三种复方中草药制剂均可显著促进罗非鱼肠道细菌、双歧杆菌、乳酸杆菌的生长(P<0.05),C方中草药制剂还可显著抑制大肠杆菌生长(P<0.05)。结果表明,C方中草药制剂可显著促进罗非鱼肠道中乳酸杆菌等有益菌群的增殖,抑制大肠杆菌生长,从而有效促进罗非鱼生长。     

长松藻基本营养成分分析及其多糖的提取

对南海长松藻(Codium cylindricum Holmes)的主要营养成分进行了分析,结果显示:多糖、蛋白质和粗纤维是构成藻体的主要营养成分,占藻体干基的80%左右,其中多糖占55.57%、粗纤维占11.09%、蛋白质占12.57%,但脂肪含量仅为0.59%。水溶性营养成分较为丰富,水浸出物总量为34.55%,其中包括水溶性蛋白、水溶性多糖、游离氨基酸等。多糖是长松藻的主要有效成分。单因素及正交实验结果表明:长松藻多糖提取的最佳工艺参数为浸提温度95℃,浸提时间1.5 h,料水质量比1∶50,多糖得率为9.79%。     

Studies on the effects on growth and antioxidant responses of two marine microalgal species to uniconazole

Uniconazole, as a plant growth retardant, can enhance stress tolerance in plants, possibly because of improved antioxidation defense mechanisms with higher activities of superoxide dismutase(SOD) and peroxidase(POD) enzymes that retard lipid peroxidation and membrane deterioration. These years much attention has been focused on the responses of antioxidant system in plants to uniconazole stress, but such studies on aquatic organism are very few. Moreover, no information is available on growth and antioxidant response in marine microalgae to uniconazole. In this paper, the growth and antioxidant responses of two marine microalgal species, Platymonas helgolandica and Pavlova viridis, at six uniconazole concentrations(0-15 mg L-1) were investigated. The results demonstrated that 3 mg L-1 uniconazole could increase significantly chlorophyll a and carbohydrate contents of P. helgolandica(P 0.05). Higher concentrations(≥12 mg L-1) of uniconazole could inhibit significantly the growth, dry weight, chlorophyll-a and carbohydrate contents of P. helgolandica and P. viridis(P 0.05). Uniconazole caused a significant increase in lipid peroxidation production(MDA) at higher concentrations(≥ 9 mg L-1). The activities of antioxidant enzymes, superoxide dismutase(SOD) and catalase(CAT) were enhanced remarkably at low concentrations of uniconazole. However, significant reduction of SOD and CAT activities was observed at higher concentrations of uniconazole.     

Production of dimethylsulfide and acrylic acid from dimethylsulfoniopropionate during growth of three marine microalgae

We measured the concentrations of dimethylsulfide （DMS）, acrylic acid （AA）, and dimethylsulfoniopropionate （DMSP） during growth of three microalgae： Prorocentrum micans, Gephyrocapsa oceanica, and Platymonas subcordiformis. The DMSP, AA, and DMS concentrations in culture media varied significantly among algal growth stages, with the highest concentrations in the late stationary growth stage or the senescent stage. In the stationary growth stage, the average DMSP concentration per cell in P. micans （0.066 5 pmol/cell） was 1.3 times that in G. oceanica （0.049 5 pmol/ cell） and 20.2 times that in P. subcordiformis （0.003 29 pmol/cell）. The average concentrations of AA were 0.044 6, 0.026 9, and 0.003 05 pmol/cell in P. micans, G. oceanica, and P. subcordiformis, respectively, higher than the concentrations of DMS （0.272, 0.497, and 0.086 2 fmol/cell, respectively）. There were significant positive correlations between cell density and AA, DMSP, and DMS concentrations. The ratios of DMS/AA and AA/（DMSP＋AA） in the three algae differed significantly over the growth cycle. In all three microalgae, the DMS/AA ratios were less than 25% during the growth period, suggesting that the enzymatic cleavage pathway, which generates DMS, was not the main DMSP degradation pathway. The changes in the DMS/AA ratio indicated that there was a higher rate of enzymatic breakdown of DMSP in the early growth period and a lower rate during senescence. In all three microalgae, the AA/（DMSP＋AA） ratio （degradation ratio of DMSP） decreased during the exponential growth phase, and then increased. The variations in these ratios can approximately indicate the cleavage mechanism of DMSP at different stages of algal growth.     

Synthesis and Characterization of a Hydroxyethyl Derivative of Chitosan and Evaluation of Its Biosafety

Hydroxyethyl chitosan(HE-chitosan) is a water-soluble derivative of chitosan with many apparent biological properties. For example, it is non-toxic and rapidly biodegradable. Moreover, HE-chitosan has advantages in water-solubility, moisture retention and gelling property due to its hydroxyethyl group. However, the biocompatibility and biodegradability of this multifunctional derivative have rarely been documented although they are critical for its application in biomedical and clinical treatments. The purpose of this work was to evaluate the biosafety of HE-chitosan, and draw important clues for its diverse applications. HE-chitosan was synthesized and characterized its chemical structure with FTIR. Its molecular weight(MW) was determined by gel permeation chromatography(GPC), and its deacetylation degree(DD) was investigated through potentiometric analysis. The cytotoxicity of HE-chitosan on mouse fibroblast cell L929 was tested. The biocompatibility and biodegradability of HE-chitosan in rat and rabbit were evaluated. The FTIR results indicated that the hydroxyethyl groups were linked to C6 of chitosan. The GPC analysis confirmed that its Mw was about 90.01 k Da. It was also demonstrated that HE-chitosan had excellent biocompatibility and biodegradability in vivo and had no cytotoxicity on L929. These findings indicated that HE-chitosan can potentially be applied as a biomaterial in tissue engineering, drug delivery, and other biomedical fields.     

Single nucleotide polymorphisms in intron 1 and intron 2 of Larimichthys crocea growth hormone gene are correlated with growth traits

The growth hormone gene (GH) affects animal growth and is a potential target for genetic studies of variation related to growth traits. In this study, we analyzed single nucleotide polymorphisms (SNPs) in GH intron regions and their associations with growth traits in large yellow croaker, Larimichthys crocea, from Zhejiang and Fujian stocks. The results of PCR-single strand conformation polymorphism showed two haplotypes of intron 1, named AA and AB genotypes, in Zhejiang stock. AB exhibited an SNP at position 196 (G A) that was negatively correlated with body height and positively correlated with standard length/body height (P 0.05). Two different genotypes, CC and CD, were identified in intron 2 in Fujian stock, with CD showing an SNP at position 692 (T C). The CD genotype had a significantly positive correlation with both weight and total length (P 0.01). These basic data highlight the potential for using GH as a genetic marker of fish growth in marker assisted selection.     

The aggregation behavior and formation of nanoparticles of oleoylchitosan in dilute aqueous solution

Oleoylchitosans (O-chitosans) with different degrees of substitution (DS) were synthesized by reacting chitosan with oleoyl chloride. The chemical structures of the products were characterized by 1H NMR and FT-IR. These results suggested the for-mation of an amide linkage between the amino groups of chitosan and the carboxyl groups of oleic acid. The viscosity of O-chitosan sharply increased with the increase of concentration, whereas that of unmodified chitosan rose only slightly. This increase became larger as the DS increased. All of the O-chitosans could reduce surface tension slightly. The critical aggregation concentration (CAC) of O-chitosans with DS 5%, 11%, and 27% were 79.43 mgL-1, 31.6 mgL-1, and 10mgL-1, respectively. Nanoparticles were prepared using an O/W emulsification method. The mean diameters of the polymeric amphiphilic nanoparticles of O-chitosans with DS 5% and 11% were around 327.4 nm and 275.3 nm, respectively.     

地衣芽孢杆菌在异育银鲫日粮中的应用

在异育银鲫基础日粮中分别添加质量分数0.01%、0.02%、0.03%、0.04%的地衣芽孢杆菌,测定了鱼体的生长及食糜植酸酶活性等。结果表明:与对照组相比,添加量0.01%、0.02%、0.03%组的增重率分别显著提高了12.46%、18.59%、18.84%;添加量0.02%、0.03%组的磷表观消化率分别提高37.63%、35.03%(P<0.05);饵料系数降低了0.52、0.57(P<0.05),其他组差异不显著。与对照组相比,0.03%组食糜植酸酶活性提高50.79%、0.04%组肝体比降低20.91%(P<0.05),其他组差异不显著。因此在异育银鲫日粮中添加地衣芽孢杆菌提高了机体酶活性,促进了鱼体生长。     

Use of tetracycline hydrochloride and alizarin complexone for immersion marking black rockfish Sebastes schlegelii

We tested the utility of chemical marking techniques in the juvenile black rockfish Sebastes schlegelii. Juveniles(30–40 mm total length) were immersed in a range of tetracycline hydrochloride(TC) solutions at concentrations ranging from 300 to 500 mg/L, and alizarin complexone(ALC) solutions at concentrations ranging from 200 to 400 mg/L in filtered sea water(salinity of 30) for 24 h, respectively. Otoliths(sagittae, asteriscus), scales, fin rays(dorsal, pectoral, ventral, anal, and caudal fin rays), and fin spines(dorsal, ventral, and anal fin spines) were sampled and used to detect fl uorescent marks after a 60-day growth experiment. With the exception of 300 mg/L TC, acceptable marks were produced in the otoliths and fin spines by all concentrations of TC and ALC. In particular, we observed clearly visible marks in the sagittae, asteriscus, and fin spines under normal light at concentrations of 200–400 mg/L, 250–400 mg/L, and 250–400 mg/L ALC, respectively. Scales and fin rays had acceptable marks at much higher concentrations(≥350 mg/L TC, ≥250 mg/L ALC for scales and ≥350 mg/L TC, ≥300 mg/L ALC for fin rays). The best mark quality(i.e., acceptable marks were observed in all sampled structures after immersion marking) were obtained following immersion in TC at between 350–500 mg/L, and ALC between 300–400 mg/L. In addition, there was no significant difference in survival and growth of TC and ALC marked fish compared to their controls up to 60 days post-marking( P 0.05).     

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.     

Strain H<Subscript>2</Subscript>-419-4 of <Emphasis Type="Italic">Haematococcus pluvialis</Emphasis> induced by ethyl methanesulphonate and ultraviolet radiation

Two strains H2-410 and H2-419 were obtained from the chemically mutated survivors of wild Haematococcus pluvialis 2 by using ethyl methanesulphonate (EMS). Strains H2-410 and H2-419 showed a fast cell growth with 13% and 20% increase in biomass compared to wild type, respectively. Then H2-419-4, a fast cell growth and high astaxanthin accumulation strain, was obtained by exposing the strain H2-419 to ultraviolet radiation (UV) further. The total biomass, the astaxanthin content per cell, astaxanthin production of H2-419-4 showed 68%, 28%, and 120% increase compared to wild H. pluvialis 2, respectively. HPLC (High Performance Liquid Chromatography) data showed also an obvious proportional variation of different carotenoid compositions in the extracts of H2-419-4 and the wild type, although no peak of carotenoids appeared or disappeared. Therefore, the main compositions in strain H2-419-4, like its wild one, were free of astaxanthin, monoester, and diester of astaxanthin. The asexual reproduction in survivors after exposed to UV was not synchronous, and different from the normal synchronous asexual reproduction as the mother cells were motile instead of non-motile. Interestingly, some survivors from UV irradiation produced many mini-spores (or gamete?), the spores moved away from the mother cell gradually 4 or 5 days later. This is quite similar to sexual reproduction described by Elliot in 1934. However, whether this was sexual reproduction remains questionable, as no mating process has been observed.     

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.     

Measurement and spatial distribution of urban agglomeration industrial compactness in China

Urban agglomeration （UA） compactness means spatial concentration degree of physical entities, such as cities （towns）, industries, resources, funds, traffic and technologies, whose concentration is formed according to specified economic and technologic association in the process of UA formation and development. The UA industrial compactness means the concentration degree of industry and industry clusters with reference to the industrial, technological and economic relations among the cities in the UA in the process of rational industrial division and with the extension of industrial chain. After analyzing the researches on compactness, this paper finds that the relevant measurement coefficient and methods reflecting industrial geographical concentration fail to link industries spatial concentration with urban spatial concentration. Taking 23 UAs as samples and classifying them by development degree, this paper probes into UA compactness and spatial distribution characteristics from the perspective of industry by adopting UA index systems of industry and measurement models. The research finds out： 1） there is obvious positive correlation between UA industrial compactness and UA development degree; 2） the spatial distribution difference of UA industrial compactness is relatively great; and 3） UA industrial compactness shows a gradually decreasing tendency from the eastern part, the middle part to the western part of China. From the research thoughts and approaches, this article suggests that studies on the UA integrated compactness measurement should be enhanced from a multidimensional per- spective involving space, traffic, population density and so on.