Chemical constituents of marine medicinal mangrove plant <Emphasis Type="Italic">Sonneratia caseolaris</Emphasis>

Twenty-four compounds including eight steroids (1–8), nine triterpenoids (9–16, 24), three flavonoids (20–22), and four benzenecarboxylic derivatives (17–19, 23) were isolated and identified from stems and twigs of medicinal mangrove plant Sonneratia caseolaris. The structures of the isolated compounds were determined by extensive analysis of their spectroscopic data. Among these metabolites, compounds 1, 4–20 and 22–24 were isolated and identified for the first time from S. caseolaris. In the in vitro cytotoxic assay against SMMC-7721 human hepatoma cells, compound 21 (3′,4′,5,7-tetrahydroxyflavone) exhibited significant activity with IC₅₀ 2.8 μg/mL, while oleanolic acid (14), 3,3′-di-O-methyl ether ellagic acid (18), and 3,3′,4-O-tri-O-methyl ether ellagic acid (19) showed weak activity. None of these compounds displayed significant antibacterial activites. Supported by the National Natural Science Foundation of China (No. 30770234); Knowledge Innovation Program of Chinese Academy of Sciences (KZCX2-YW-211-04); Department of Science and Technology of Shandong Province (No.2006GG2205023)     

The antitumor effect of bromophenol derivatives in vitro and Leathesia nana extract in vivo

To investigate the antitumor effect of bromophenol derivatives in vitro and Leathesia nana extract in vivo, six bromophenol derivatives 6-(2,3-dibromo-4,5-dihydroxybenzyl)-2,3-dibromo-4,5-dihydroxy benzyl methyl ether (1), (+)-3-(2,3-dibromo-4,5-dihydroxyphenyl)-4-bromo-5,6-dihydroxy-1,3-dihydroisobenzofuran (2), 3-bromo-4-(2,3-dibromo-4,5-dihydroxybenzyl)-5-methoxymethyl-pyrocatechol (3),2,2',3,3'-tetrabromo-4,4',5,5'-tetrahydroxy-diphenylmethane (4), bis(2,3-dibromo-4,5-dihydroxybenzyl) ether (5), 2,2',3-tribromo-3',4,4',5-tetrahydroxy-6'-ethyloxymethyldiphenylmethane (6) were isolated from brown alga Leathesia nana, and their cytotoxicity were tested by MTr assays in human cancer cell lines A549, BGC-823, MCF-7, B16-BL6, HT-1080, A2780, Bel7402 and HCT-8. Their inhibitory activity against protein tyrosine kinase (PTK) with over-expression of c-kit was analyzed also by ELISA. The antitumor activity of ethanolic extraction of Leathesia nana (EELN) was evaluated on S180-bearing mice.All compounds showed very potent cytotoxicity against all of the eight cancer cell lines with IC50 below 10 μg/mL. In PTK inhibition study, all bromophenol derivatives showed moderate inhibitory activity and compounds 2, 5 and 6 showed significant bioactivity with the inhibition ratio of 77.5%, 80.1% and 71.4%,respectively. Pharmacological studies reveal that EELN could inhibit the growth of Sarcoma 180 tumor and increase the indices of thymus and spleen to improve the immune system remarkably in vivo. Results indicated that the bromophenoi derivatives and EELN can be used as potent antitumor agents for PTK over-expression of c-kit and considered in a new therapeutic strategy for treatment of cancer.     

The Antitumor Components from Marine-derived Bacterium Streptoverticillium luteoverticillatum 11014 Ⅱ

Eight known compounds were isolated from a marine-derived bacterium Streptoverticillium luteoverticillatum 11014 using bioassay-guided fractionations. Their structures were identified by spectral analysis as bis (4-hydroxybenzyl) ether (1), p-hydroxyphenylethyl alcohol (2), N-(4-hydroxyphenethyl) acetamide (3), indole-3 carboxylic acid methyl ester (4), dibenzo[b,e] [1,4]dioxine (5), thymine (6), cytosine deoxyribonucleoside (7) and 2, 3-butanediol (8). These compounds were evaluated for their cytotoxic activities against K562 cell line with the SRB method for the first time. Compounds 2 and 4 showed cytotoxcities with IC50 values of 101.1 and 165.3 μmolL-1, respectively. All compounds were isolated from S. luteoverticillatum 11014 for the first time.     

Chemical constituents of marine algal-derived endophytic fungus Exophiala oligosperma EN-21

Seven compounds (1–7) were identified from the cultivation of the endophytic fungus Exophiala oligosperma (EN-21) that was isolated from the inner tissue of the marine red alga Laurencia similis. Their structures were identified with spectroscopic and chemical methods as 2-phenoxynaphthalene (1), (2S, 3R, 4E, 8E)-1-O-β-D-glucopyranosyl-3-hydroxy-2-[(R)-2′-hydroxyoctadecanoyl] amino-9-methyl-4, 8-octadeca-diene (2), (22E,24R)-ergosta-7,22-dien-3β,5α,6β-triol (3), (22E, 24R)-3β, 5α, 9α-trihydroxy- ergosta-7, ...     

The Antitumor Components from Marine-derived Bacterium Streptoverticillium luteoverticillatum 11014 Ⅱ

Eight known compounds were isolated from a marine-derived bacterium Streptoverticillium luteoverticillatum 11014 using bioassay-guided fractionations. Their structures were identified by spectral analysis as bis （4-hydroxybenzyl） ether （1）, p-hydroxyphenylethyl alcohol （2）, N-（4-hydroxyphenethyl） acetamide （3）, indole-3 carboxylic acid methyl ester （4）, dibenzo[b,e] [1,4]dioxine （5）, thymine （6）, cytosine deoxyribonucleoside （7） and 2, 3-butanediol （8）. These compounds were evaluated for their cytotoxic activities against K562 cell line with the SRB method for the first time. Compounds 2 and 4 showed cytotoxcities with IC50 values of 101.1 and 165.3 μolL^-1, respectively. All compounds were isolated from S. luteoverticillatum 11014 for the first time.     

Extraction and PTP1B inhibitory activity of bromophenols from the marine red alga Symphyocladia latiuscula

Previously, we had characterized several structurally interesting brominated phenols from the marine red alga Symphyocladia latiuscula collected from various sites. However, Phytochemical investigations on this species collected from the Weihai coastline of Shandong Province remains blank. Therefore, we characterized the chemical constituents of individuals of this species collected from the region. Eight bromophenols were isolated and identified. Using detailed spectroscopic techniques and comparisons with published data, these compounds were identified as 2,3-dibromo-4,5-dihydroxybenzyl methyl ether (1), 3,5-dibromo-4-hydroxybenzoic acid (2), 2,3,6-tribromo-4,5-dihydroxymethylbenzene (3), 2,3,6-tribromo-4,5-dihydroxybenzaldehyde (4), 2,3,6-tribromo-4,5-dihydroxybenzyl methyl ether (5), bis(2,3,6-tribromo-4,5-dihydroxyphenyl)methane (6), 1,2-bis(2,3,6-tribromo-4,5-dihydroxyphenyl)-ethane (7), and 1-(2,3,6-tribromo-4,5-dihydroxybenzyl)-pyrrolidin-2-one (8). Among these compounds, 1 and 2 were isolated for the first time from S. latiuscula. Each compound was evaluated on the ability to inhibit protein tyrosine phosphatase 1B (PTP1B), which is a potential therapeutic target in the treatment of type 2 diabetes. Bromophenols 5, 6, and 7 showed strong activities with IC50 values of 3.9, 4.3, and 3.5 μmol/L, respectively. This study provides further evidence that bromophenols are predominant among the chemical constituents of Symphyocladia, and that some of these compounds may be candidates for the development of anti-diabetes drugs.     

The antitumor effect of bromophenol derivatives <Emphasis Type="Italic">in vitro</Emphasis> and <Emphasis Type="Italic">Leathesia nana</Emphasis> extract <Emphasis Type="Italic">in vivo</Emphasis>

To investigate the antitumor effect of bromophenol derivatives in vitro and Leathesia nana extract in vivo, six bromophenol derivatives 6-(2,3-dibromo-4,5-dihydroxybenzyl)-2,3-dibromo-4,5-dihydroxy benzyl methyl ether (1), (+)-3-(2,3-dibromo-4,5-dihydroxyphenyl)-4-bromo-5,6-dihydroxy-1,3-dihydroisobenzofuran (2), 3-bromo-4-(2,3-dibromo-4,5-dihydroxybenzyl)-5-methoxymethyl-pyrocatechol (3), 2,2′,3,3′-tetrabromo-4,4′,5,5′-tetrahydroxy-diphenylmethane (4), bis(2,3-dibromo-4,5-dihydroxybenzyl) ether (5), 2,2′,3-tribromo-3′,4,4′,5-tetrahydroxy-6′-ethyloxymethyldiphenylmethane (6) were isolated from brown alga Leathesia nana, and their cytotoxicity were tested by MTT assays in human cancer cell lines A549, BGC-823, MCF-7, B16-BL6, HT-1080, A2780, Bel7402 and HCT-8. Their inhibitory activity against protein tyrosine kinase (PTK) with over-expression of c-kit was analyzed also by ELISA. The antitumor activity of ethanolic extraction of Leathesia nana (EELN) was evaluated on S₁₈₀-bearing mice. All compounds showed very potent cytotoxicity against all of the eight cancer cell lines with IC₅₀ below 10 μg/mL. In PTK inhibition study, all bromophenol derivatives showed moderate inhibitory activity and compounds 2, 5 and 6 showed significant bioactivity with the inhibition ratio of 77.5%, 80.1% and 71.4%, respectively. Pharmacological studies reveal that EELN could inhibit the growth of Sarcoma 180 tumor and increase the indices of thymus and spleen to improve the immune system remarkably in vivo. Results indicated that the bromophenol derivatives and EELN can be used as potent antitumor agents for PTK over-expression of c-kit and considered in a new therapeutic strategy for treatment of cancer. Supported by the National High Technology Research and Development Program of China (863 Program, No. 2007AA09Z410) and Knowledge Innovation Program of Chinese Academy of Sciences (No. KZCX2-YW-209)     

盐酸普鲁卡因诱导下海洋真菌Hypocrea lixii次级代谢化学成分

【目的】通过化学诱导方法从一株海洋真菌Hypocrea lixii中获得多样化的活性小分子化合物。【方法】对菌株在盐酸普鲁卡因的化学诱导作用下进行固体发酵,代谢产物采用硅胶减压柱层析、Sephadex LH-20以及pHPLC等手段分离纯化,对化合物运用NMR、MS等波谱学技术并比对文献进行结构鉴定,采用Ellman比色法及DPPH自由基清除法对化合物进行初步抗乙酰胆碱酯酶(AChE)和抗氧化活性测试。【结果】从海洋真菌Hypocrea lixii在盐酸普鲁卡因诱导下的发酵提取物中分离得到4个异黄酮类化合物,包括6'-O-crotonylgenistin(1)、Genistein(2)、Daidzein(3)和Genistin(4),以及对氨基苯甲酸甲酯(5)、对羟基苯丙酸(6)和胞嘧啶(7),其中化合物1和6为该种中首次报道,化合物5为生物转化来源首次报道,化合物2~4可能源自培养基。在100μmol/L剂量浓度下,化合物1~4对AChE的抑制以及DPPH自由基的清除活性均较弱,化合物5和6显示了相对强的AChE抑制活性,抑制率分别为40%和38%。【结论】盐酸普鲁卡因对该菌次级代谢产生了显著的影响,为深入研究该菌株化学诱导次级代谢产物提供基础。     

The Cytotoxic Constituents from Marine-derived Streptomyces 3320^#

The present work studies the chemical constituents from marine-derived streptomyces 3320^# and their antitumor activities. The n-BuOH extract of the ferment broth of 3320^# was chromatographed on silica gel, Sephadex LH-20, ODS columns and HPLC to separate the compounds with antitoumor activities. Their structures were identified using IR, UV, NMR, MS spectroscopic techniques and compared with published data. The antitumor activities of the isolates were assayed using SRB method and flow cytometry assay, accompanied with the morphological observation of the cells under light micro- scope against mammalian tsFT210 cells. Ten compounds, cyclo-（Ala-Leu） 1, cyclo-（Ala-Ile） 2, cyclo-（Ala-Val） 3, cyclo- （Phe-Pro） 4, cyclo-（Phe-Gly）5, cyclo-（Leu-Pro）6, 1-methyl-1, 2, 3, 4-tetrahydro-β-carboline-3-carboxylic acid 7, N-（4- hydroxyphenethyl） acetamide 8, 4-methyoxy-l-（2-hydroxy） ethylbenzene 9 and uridine 10, were isolated from the ferment broth of streptomyces 3320^# . Among them, compounds 6, 7, 8 and 10 showed potent cytotoxicity against the tsFT210 cell with the IC50 values of 3.6, 7.2, 5.2 and 1.6 mmol L-1, respectively. Compounds 8, 10 also exhibited apoptosis inducing activity under 2.0 mmol L-1. Compounds 6, 7, 8 and 10 are the principle bioactive constituents responsible for the antitumor activities of marine streptomyces 3320^#. Compound 7 was isolated from this species for the first time.     

Lipid oxidation and fatty acid composition in salt-dried yellow croaker (<Emphasis Type="Italic">Pseudosciaena polyactis</Emphasis>) during processing

Lipid oxidation in salt-dried yellow croaker (Pseudosciaena polyactis) was evaluated during processing with commonly used analytical indices, such as the peroxide value (POV), the thiobarbituric acid reactive substances (TBARS) value, and oxidative-relative lipoxygenase (LOX) activity. Additionally, fatty acids were analyzed using gas chromatography-mass spectrometry. Both POV and TBARS increased significantly (P < 0.05) at the rinsing stage. POV reached its peak value of 3.63 meq O₂ per kg sample at the drying stage, whereas TBARS constantly increased from 0.05 to 0.20 mg MDA per kg sample. Processing of salt-dried yellow croaker had an extremely significant (P < 0.01) effect on LOX activity. Twenty-six fatty acids were identified. Combined eicosapentaenoic acid (EPA; C20:5n3) and docosahexaenoic acid (DHA; C22:6n3) content varied between (19.20 ± 0.37) mg g^?1 and (23.45 ± 1.05) mg g^?1. The polyunsaturated fatty acid/saturated fatty acid (PUFA/SFA) ratio in yellow croaker was 0.73–1.10, and the n-6/n-3 PUFA ratio was approximately 0.13–0.20. The contents of most fatty acids varied significantly (P < 0.05) during the different processing stages, and these differences were caused by lipid oxidation. C18:0, C16:1n7, C19:0, and C22:6n3 showed clear changes in principle component one of a principle components analysis. These fatty acids are potential markers for evaluating lipid oxidation in fish muscle because there was a significant correlation between these markers and TBARS and LOX activity (P < 0.05) with Pearson’s coefficients > 0.931.     

Isolation and pharmacological activities of bromophenols from Rhodomela confervoides

Four bromophenols were isolated from the extract of marine red alga, Rhodomela confervoides by column chromatography and HPLC methods. By means of spectroscopic methods including IR, MS, 1D, and 2D-NMR techniques, their structures were elucidated as (1) 3-bromo-4,5-dihydroxy benzoic acid methyl ester; (2) his (2,3-dibromo-4,5-dihydroxybenzyl) ether; (3) 3,4-dibromo-5-(methoxymethyl)-1,2-benzenediol and (4)3-bromo-4,5-dihydroxy-benzaldehyde. Compound 1 was first isolated from the algae in nature, and 1, 4 were found to have selective cytotoxic activities against KB, Bel 7402 and A549 cells, 2 showed powerful antibacterial activities against Staphylococcus aurens and Pseudomonas aeruginosa.     

The chemical constituents from red alga Gymnogongrus flabelliformis Harv.

Eight compounds were isolated from red alga Gymnogongrus flabelliformis Harv. In normal phase silica gel, Sephadex LH-20 gel column chromatography, reverse phase HPLC, and recrystallization. Based on MS and ID NMR spectroscopic data, their structures were determined as： stigmast-4-en-3-one （Ⅰ）, cholest-4-en-3-one （Ⅱ）, cholesterol （m）, uracil （Ⅳ）, uridine （V）, adenosine （Ⅵ）, succinic acid （Ⅶ）, and 5-hydroxy-4-methyl-5-pentyl-2,5-dihydro-furan-2-on （Ⅷ）. All of them were obtained from this species for the first time. Cytotoxicity of these compounds was screened using standard MTT method, but all the compounds were inactive （IC50 〉 10 μg/ml）.     

Isolation and pharmacological activities of bromophenols from Rhodomela confervoides

Four bromophenols were isolated from the extract of marine red alga,Rhodomela confervoides by column chromatography and HPLC methods. By means of spectroscopic methods inclding IR, MS, 1D, and 2D-NMR techniques, their structures were elucidated as (1) 3-bromo-4,5-dihydroxy benzoic acid methyl ester; (2) bis (2,3-dibromo-4,5-dihydroxybenzyl) ether; (3) 3,4-dibromo-5-(methoxymethyl)-1,2-benzenediol and (4) 3-bromo-4,5-dihydroxy-benzaldehyde. Compound 1 was first isolated from the algae in nature, and 1, 4 were found to have selective cytotoxic activities against KB, Bel 7402 and A549 cells, 2 showed powerful antibacterial activities againstStaphylococcus aurens andPseudomonas aeruginosa. Supported by National “863” Program (No. 2004AA625030, 2001AA620503), Quingdao marine sciences project (No. 04-2-NN-26) and Key Knowledge Innovative Project of the Chinese Academy of Sciences (KZCX3-SW-215).     

流沙湾的底栖大型海藻调查

2008年3月～2009年1月调查了流沙湾潮间带及湾内(109°55'～109°59'E,20?22'～20°28'N)底栖大型海藻资源,定性、定量分析该海湾底栖大型海藻种类、生态分布、区系性质、群落结构及生物量的季节变化规律。共采集底栖大型海藻3门10目17科19属32种,其中红藻门4目9科9属15种,绿藻门4目6科7属14种,褐藻门2目2科3属3种,以亚热带藻种占绝对优势,达59.4%,温带成分21.9%,热带成分18.7%。采样海藻具印度-西太平洋海洋植物区系中国-日本亚热带海洋植物亚区特点。调查区域内底栖大型海藻群落组成的季节变化明显。     

Algicidal activities of secondary metabolites of marine macroalgal-derived endophytic fungi

Red tide occurrs frequently and causes signi?cant damage to the environment and human health. As a result, development of new effcient and environment friendly red-tide microalgae inhibitors has gained increasing attention in recent times. Algicolous endophytic fungi with unique habitats are promising sources for active agents owing to their abundant secondary metabolites and distinguished activities. In this study, the algicidal activities of 49 marine macroalgal-derived endophytic fungi against phytoplankton Alexandrium tamarense, Prorocentrum donghaiense, Heterosigma akashiwo, and Chattonella marina were examined using 96-well microplate. Four fungal strains, including Aspergillus wentii(pt-1), A. ustus(cf-42),and A. versicolor(dl-29, pt-20), exhibited potent algicidal activities. A total of 32 pure compounds isolated from these fungi were noted to possess dif ferent degrees of algicidal activities. Of those, 11 compounds comprising ?ve anthraquinones, two terpenoids, and four steroids showed high 24-h inhibition rates for the four red tide algae, with 24 h EC_(50) values ranging from 0.01 to 14.29 μg/mL. Among them, compound1(1,5-dihydroxy-3-methoxy-7-methylanthraquinone) presented the strongest activity against H. akashiwo,and could decrease its chlorophyll a(Chl a) and superoxide dismutase contents and increase the soluble protein, malondialdehyde, and peroxidase contents. These results suggested that the identi?ed anti-algal compound might inhibit the growth of red tide algae by weakening photosynthesis(reducing Chl a content),destroying cell membrane, and damaging the antioxidant system.     

一株溶藻弧菌噬菌体的生理特性研究

对从湛江市水产品批发市场分离得到一株溶藻弧菌噬菌体的生理特性进行测定,结果表明:该株噬菌体噬菌斑12 h直径为1～1.5 mm,对紫外线敏感,暴露在紫外灯(20 W,30 cm)下30 min可丧失全部裂解活性;在pH8～11范围内裂解活性较强,最适pH值为11,裂解周期为65 min,对65℃以上的高温敏感,对乙醚和氯仿有较好的耐受性,对抗病毒药物阿昔洛韦敏感,最大常规稀释度为106CFU/mL,最佳感染复数为10。     

温度对匙吻鲟胚胎与仔鱼发育的影响

研究不同温度下匙吻鲟(Polyodon spathula)的胚胎与仔鱼发育。结果表明,26℃时,匙吻鲟胚胎发育到大卵黄栓期全部死亡;在15～24℃范围内,胚胎均可孵化出膜,孵化时间与温度呈极显著的负相关关系(P<0.01)。胚胎发育的最适温度范围为18～22℃,最佳温度为20℃,生物学零度为12.30℃。卵黄囊期仔鱼历时随温度的升高而缩短。在16～22℃范围内,温度对卵黄囊期仔鱼生长的影响极显著(P<0.01),22～24℃影响不显著(P<0.05)。在22～26℃范围内,仔鱼生长的差异不显著(P<0.05),成活率低,且畸形率极显著地高于16～20℃(P<0.01)。匙吻鲟仔鱼早期发育的最适温度范围为18～20℃,温度过高是造成仔鱼畸形率增加的主要原因之一。     

Chemical constituents and antioxidant activities of waste liquid extract from Apostichopus japonicus Selenka processing

Apos tichopus japonicus Selenka is an ideal tonic food that is used traditionally in many Asian countries, and it contains many bioactive substances, such as antioxidant, antimicrobial, and anticancer materials. To convert waste liquid generated during production into a useful resource, extract from waste liquid was isolated by column chromatography and studied by the pyrogallol autoxidation and 1,10-phenanthroline-Fe²⁺ oxidation methods. Results show that the extract scavenged about 91% of the superoxide anion radical at a concentration of 1.4 mg/mL and 24% of the hydroxyl radical at 3.3 mg/mL. Four compounds were isolated and identified from the extract: 2,4-dihydroxy-5-methyl-1,3-azine; 2,4-dihydroxy-1,3-diazine; 3-O-[β-D-quinovopranosyl-(1→2)-4-O-sodium sulfate-β-D-xylopranosyl]-holosta-9(11)-ene-3β,12α,17α-triol; and 24-ethyl-5α-cholesta-7-ene-3β-O-β-D-xylopyranoside. All of these compounds are known in A. japonicus, and were found in the waste liquid for the first time.     

Steroids from green alga <Emphasis Type="Italic">Chaetomorpha basiretorsa</Emphasis> Setchell

Six steroids have been isolated from ethanolic extract of green alga Chaetomorpha basiretorsa Setchell by a combination of repeated normal phase silica gel and Sephadex LH-20 gel column chromatography as well as recrystallization. Using spectroscopic methods including MS and NMR, their structures were determined as β-1awsaritol （1）, saringosterol （2）, 24-hydroperoxy-24-vinyl- cholesterol （3）, β-stigmasterol （4）, stigmast-4-en-3α, 6β-diol （5）, 29-hydroxystigmasta-5, 24 （28）-dien-3β-ol （6）. All these compounds were obtained from this genus for the first time and they were inactive （IC50〉10μg/ml） against KB, Bel-7402, PC-3M, Ketr 3 and MCF-7 cell lines.     

流沙湾贝类资源调查

对流沙湾潮间带及湾内浅水地带(20°22'103″～20°28'526″N,109°55'341″～109°59'185″E)的贝类资源进行周年调查,定性、定量分析该海湾贝类种类、生态分布、区系性质、群落结构及生物量的季节变化规律。结果表明:在流沙湾采集到的贝类有3纲7亚纲13目42科79属104种,其中帘蛤科24种,牡蛎科、贻贝科各6种,汇螺科和蜒螺科各5种,其他各科均小于5种;贝类组成表现出温带性质,温带种类占绝对优势,该海湾贝类区系属印度—西太平洋的中国-日本亚区,生物量的季节变化明显。