浅论海洋生物医药产业发展现状、趋势与建议

随着海洋生物科技的发展，世界各国对相关成果转化力度加大，从而为海洋生物医药相关科技的成熟与发展提供了新的契机。中国海洋生物医药资源丰富，并且拥有独特的传统中医药理体系，为海洋生物医药的发展提供了良好的基础。目前，海洋药物研发进入海洋天然产物与新药重点开发并举的时期，多重资本形式开始介入，推动海洋生物医药产业实现了25％～30％的高速增长，该产业有望成为战略性的支柱产业。因此，在政策支持、技术创新和投资力度方面，应有针对性地加强对海洋生物医药产业的支持，为海洋生物医药产业的健康发展提供良好基础。

Promotive Effects of Alginate-Derived Oligosaccharides on the Inducing Drought Resistance of Tomato

In order to determine the role of alginate-derived oligosaccharides (ADO) in drought stress resistance of tomato (Ly-copersicon esculentum Miller) seedlings, the leaves were exposed to different concentrations of ADO (0.05%, 0.10%, 0.20%, 0.30% and 0.50%) after drought stress was simulated by exposing the roots to 0.6 molL-1 PEG-6000 solution for 6 h. Changes in biomass, electrolyte leakage and malondialdehyde (MDA), free proline, total soluble sugars (TSS) and abscisic acid (ABA), the enzyme activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD) and phenylalanine ammonia-lyase (PAL) were measured to investigate the effects of ADO treatment. The results showed that the treatment with an ADO concentration of 0.20% exhibited the highest performance of drought stress resistance in the tomato seedlings by decreasing the electrolyte leakage and the concentration of MDA, increasing the contents of free proline, TSS and ABA, and increasing the activities of CAT, SOD, POD and PAL after treatment with ADO. It is suggested that changes in electrolyte leakage, MDA, osmotic solutes, ABA, anti-oxidative enzyme and PAL activities were responsible for the increased drought stress resistance in tomato seedlings. To our best knowledge, this is the first report of the effect of ADO treatment on enhancing the drought stress resistance of tomato seedlings.

Efficient gusA Transient Expression in Porphyra yezoensis Protoplasts Mediated by Endogenous Beta-tubulin Flanking Sequences

Endogenous tubulin promoter has been widely used for expressing foreign genes in green algae, but the efficiency and feasibility of endogenous tubulin promoter in the economically important Porphyra yezoensis (Rhodophyta) are unknown. In this study, the flanking sequences of beta-tubulin gene from P. yezoensis were amplified and two transient expression vectors were con-structed to determine their transcription promoting feasibility for foreign gene gusA. The testing vector pATubGUS was constructed by inserting 5’- and 3’-flanking regions (Tub5’ and Tub3’) up- and down-stream of β-glucuronidase (GUS) gene (gusA), respectively, into pA, a derivative of pCAT?3-enhancer vector. The control construct, pAGUSTub3, contains only gusA and Tub3’. These con-structs were electroporated into P. yezoensis protoplasts and the GUS activities were quantitatively analyzed by spectrometry. The results demonstrated that gusA gene was efficiently expressed in P. yezoensis protoplasts under the regulation of 5’-flanking sequence of the beta-tubulin gene. More interestingly, the pATubGUS produced stronger GUS activity in P. yezoensis protoplasts when com-pared to the result from pBI221, in which the gusA gene was directed by a constitutive CaMV 35 S promoter. The data suggest that the integration of P. yezoensis protoplast and its endogenous beta-tubulin flanking sequences is a potential novel system for foreign gene expression.

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.

Preparation, Characterization and Pharmacokinetics of Fluorescence Labeled Propylene Glycol Alginate Sodium Sulfate

A rapid and sensitive fluorescence labeling method was developed and validated for the microanalysis of a sulfated polysaccharide drug,namely propylene glycol alginate sodium sulfate(PSS), in rat plasma. Fluorescein isothiocyanate(FITC) was selected to label PSS, and 1, 6-diaminohexane was used to link PSS and FITC in order to prepare FITC-labeled PSS(F-PSS) through a reductive amination reaction. F-PSS was identified by UV-Vis, FT-IR and 1H-NMR spectrum. The cell stability and cytotoxicity of F-PSS were tested in Madin-Darby canine kidney(MDCK) cells. The results indicated that the labeling efficiency of F-PSS was 0.522% ± 0.0248% and the absolute bioavailability was 8.39%. F-PSS was stable in MDCK cells without obvious cytotoxicity. The method was sensitive and reliable; it showed a good linearity, precision, recovery and stability. The FITC labeling method can be applied to investigating the absorption and metabolism of PSS and other polysaccharides in biological samples.

What Depth Should Deep-Sea Water be Pumped up from in the South China Sea for Medicinal Research?

In this study, seawater was pumped up from 150, 200, 300, 500 and 1000 m in the South China Sea and analyzed to make certain what depth should deep-sea water (DSW) be pumped up for medicinal usage. The pumping depth of DSW was determined on the basis of chemical ingredients. The analyses of inorganic elements and dissolved organic matter (DOM) were performed by inductively coupled plasma mass spectrometry (ICP-MS) and ultra performance liquid chromatography-mass spectrometry (UPLC-MS) respectively. The raw data were used for hierarchical cluster analysis (HCA) and principal component analysis (PCA). The results showed that seawater pumped up from 500 m and 1000 m was similar in their chemical ingredients, and was different from the seawater pumped up from other depths. These results indicated that seawater from more than 500 m depth had relatively stable chemical ingredients and could be used as DSW in the South China Sea.

Isolation of Protoplasts from Undaria pinnatifida by Alginate Lyase Digestion

The aim of this study is to isolate protoplasts from Undaria pinnatifida. Protoplasts of the alga were isolated enzymatically by using alginate lyase, which was prepared by fermenting culture of a strain Vibrio sp. 510. Monofacterial method was applied for optimizing digestion condition. The optimum condition for protoplast preparation is enzymatic digestion at 28 ℃ for 2 h using alginate lyase at the concentration of 213.36 U (8 mL) every 0.5 g fresh thalline with NaCl 50 and at the shaking speed of 150 r min -1 during digestion. The protoplast yield can reach 2.62±0.09 million per 0.5 g fresh leave under the optimum condition. The enzyme activity is inhibited by Ca 2+ and slightly enhanced by Fe 2+ and Mn 2+ at concentrations of 0.05, 0.08 and 0.10 mol L -1.

Synthesis and Pharmacokinetic Property Improvement of Deuterated Plinabulin 9

Plinabulin,a potent microtubule-targeting agent,is derived from marine natural diketopiperazine ‘phenylahistin'.To develop novel plinabulin analogue that could display better pharmacokinetic properties and less side effects,deuterated plinabulin 9 was synthesized and evaluated in vitro and in vivo.In comparison with plinabulin,in vivo pharmacokinetic studies indicated that the deuterated derivative 9 could alter blood circulation behavior obviously,which was proved by increased area under the plasma concentration-time curve(AUC_(0–∞)),reduced clearance(CL),and prolonged total body mean residence time(MRT).The derivative 9 also has higher inhibition rates against Bx PC-3,Jurkat and A-431 tumor cell lines as compared with its prototype plinabulin.Therefore,the deuterated compound 9 might be developed as a potential agent for different cancer treatments.

Ab Initio Prediction of ^(29)Si-NMR Chemical Shifts

The ability of several ab initio models to predict experimental 29Si-NMR chemical shift is examined. The shielding values of trimethylsilyl chloride (A), t-butyldimethylsilyl chloride (B) and allyltrimethylsilane (C) are calculated by GIAO , CSGT and IGAIM methods, using HF/6-31G*, B3LYP/6-31G*, HF/6-311+G **, B3LYP/6-311+G ** and MPW1PW91/6-311+G ** models respectively. The 29Si chemical shifts calculated by GIAO method using HF/6-311+G ** model are highly in agreement with those obtained experimentally. All of the models above reproduce the trends of chemical shifts in all cases studied, suggesting that the models are of practical value.

The influence of dihydropyridines calcium antagonists on 5-HT-induced intracellular calcium signal

Confocal laser scanning microscopy (CLSM) was applied to detect the intracellular [Ca2+] variety of fluorescent intension, with Fluo-3/AM fluorescence loaded in SFSMC. The results show that 10 μmol/L Lacidipine can reduce the frequence which 10 μmol/L 5-HT induced [Ca2+] spark in SFSMC of calcium over loading to 50%, and amplitude to 50% or so. We can draw a conclusion that dihydropyridines cal-cium antagonists lacidipine can antagonize the release of intracellular [Ca2+] which 5-HT-induced in dose dependent manner.