1.Study on the extraction and purification of glycosaminoglycans from Rhopilema esculenta and its antihyperlipidemia effects
Xiaoshi JIN ; Hongmian WU ; Min ZHONG ; Xiaoling LEI
Chinese Journal of Marine Drugs 1994;0(04):-
Objective To study the extraction and purification of glycosaminoglycans from Rhoqilema esculenta Kishinouye,optimize the enzymolysis process,and its antihyperlipidemia effects were studied.Methods Extraction of glycosaminoglycans(GAG) from Rhopilema esculenta Kishinouye by hydrolyzation method with papain,subtilisin and trypsin neutral proteinase,ethanol precipitation.The rude product of glycosaminoglycans(GAG) was purified and fractionated by the methods of adsorption,dialysis,ethanol precipitation and CTAB precipitation.The experimental hyperlipidemia mice induced by high fat forage were studied.The total cholesterol(TC) and triglyceride(TG) levels were determined.Results Optimum enzymolysis process: papain neutral proteinase1.5%,subtilisin neutral proteinase1.5%,treated for 5h;The result showed that the levels of TC and TG decreased to 6.83?1.09 mmol?L-1(P
2.Studies on antitumor activities of phycoerythrin from Gracilaria lemaneiftheormis
Meizhen CHEN ; Anshan GE ; Jie YU ; Wenjie YANG
Chinese Journal of Marine Drugs 1994;0(04):-
Objective To investigate the antitumor effects of phycoerythrin from Gracilaria lemaneiftheormis.Methods The coarse phycoerythrin(CPE) was ig administered to sarcoma180 mice,then the antitumor effect of CPE in vivo was evaluated by the inhibition rate,thymus and spleen indexes.The growth inhibitory effect of phycoerythrin(PE) on cell lines HeLa,EC109,and vascular endothelial in vitro was examined with MTT assay.Influences on HeLa cell cycle and apoptosis of PE were observed by flow cytometry and annexin v-fitc/PI fluorescence staining.Results CPE exhibited inhibitory effect against sarcoma180 growth in mice at the dosage of 100~300 mg?kg-1with a fair dose-effect relationship,and the inhibitory rate was 49.23% at the dosage of 300 mg? kg-1.CPE could also obviously increase the weight of thymus;PE could obviously inhibit the growth of HeLa cell in a dose-effect relationship,the inhibitory rate could reach more than 70%.It could prevent the transforming of HeLa cell cycle from G2/M to S phase to cause apoptosis.But PE couldn't inhibit the growth of EC109 cell,while some growth stimulating effects of PE on vascular endothelial cell were observed.Conclusion The PE from Gracilaria lemaneiftheormis could observably inhibit sarcoma180 growth in vivo.Effects of PE on different cells in vitro were differ,but showed potent inhibitory effect to HeLa cell.
3.Preliminary study on the antiultraviolet irradiation active substances of mycosporine-like amino acids from four kinds of Antarctic ice microalgae
Ning DU ; Tianxiang GAO ; Jinlai MIAO ; Botao ZHANG ; Shouqiang WANG ; Bin WANG
Chinese Journal of Marine Drugs 1994;0(04):-
Objective To study on bioactive chemical compounds against ultraviolet irradiation in antarctic ice microalgae.Methods Four kinds of Antarctic ice microalgae were cultured under the conditions of low temperature,illumination,UV-A and UV-B irradiation.The chemical compounds were extracted by methanol and analyzed by spectrophotometer and HPLC/MS.The molecular structures were elucidated according to these data.The chemical compounds were irradated by sunshine to observe the speed of degradation and to estimate the bioactivity.Results There were five active compounds in Chlorophyceae L-4 and 7 active compounds in Diatom HF-9,in which 6 compounds belong to MAAs(mycosporine-like amino acids) and were of more bioactivity against ultraviolet irradiation.Conclusion MAAs from of Antarctic ice microalgae play an important role in resisting ultraviolet irradiation.
4.Studies on the structural features and anticoagulant activity of the polysaccharide from Aeodes orbitosa
Shunchun WANG ; Songshan SHI ; Yingqi WENG ; Zhengtao WANG ; Zhibi HU
Chinese Journal of Marine Drugs 1994;0(04):-
Objective To study the structural features and anticoagulant activity of the polysaccharide isolated from Aeodes orbitosa.Methods Sugar composition analysis,methylation analysis and IR were used to characterize the structural features.The anticoagulant activity of the polysaccharide was evaluated by cutting tail and capillary methods.Results The polysaccharide was composed of 3,6-anhydrous-galactose,6-methyl-galactose,2-methyl-galactose,galactose,xylose and glucose in the molar ratios of 6.4∶0.9∶5.8∶1.6∶84.1∶1.2.The content of Sulfate group was 37.5%. The main linkage model of the polysaccharide were 1,3 and 1,4 linkages,branch point located at O-2 and 6 of galactose residues.Sulfate group located at 2 and 6 position of 1,4 linked galactose residue and 2 position of 1,3 linked galactose residue.The polysaccharide showed significant activities to extend bleeding time and coagulation time in mice.Conclusion The polysaccharide from Aeodes orbitosa was a sulfate galactan with strong anticoagulant activity.
5.Construction of the fusion protein between EGFP and IgG-binding peptide
Hongming YANG ; Yijun ZHANG ; Peng GAO
Chinese Journal of Marine Drugs 2001;0(05):-
Objective Making the fusion protein of IgG-binding peptide with enhanced green fluorescent protein(EGFP) and determining its bioactivity.Methods The enhanced green fluorescent protein(EGFP) gene was cloned into pEZZ 18 vector containing ZZ peptide gene to construct expression vector pSpA-EGFP-His.The fusion protein was expressed in E.coliDH5? and its bioactivity was detected by competitive ELISA and fluorescence properties.Results The fusion protein migrated at approximately 42kD in SDS-PAGE,which correspond to the theoretical molecular weight.The spectra of SpA-EGFP fusion protein was similar to what was reported.SpA-EGFP competed with SpA-Peroxidase to bind IgG.Conclusion The plasmid pSpA-EGFP-His correctly expressed in E.coli.The fusion protein retains the bifunctional effects of EGFP and IgG-binding activity.
6.Fluorimetric method for determination of trace lead in alginate sodium
Weili LU ; Jiachao XU ; Xin GAO ; Changhu XUE
Chinese Journal of Marine Drugs 2001;0(05):-
Objective To establish a method for determination of trace lead in alginate sodium.Method The lead in the samples was determined by fluorescence spectroscopy after been digested by hydrothermal decomposition.Results The detection limit of lead was 2.71?10-2?g?mL-1.The relative standard deviation of the three samples were 4.06%,1.57% and 2.12% respectively,the average recovery was 88.32%~100.8%.Conclusion The method had the advantages of simple operation,higher precision,higher sensitivity and repeatability and was suitable for the determination of trace lead in the alginate sodium
7.Best protecting dosage of phycocyanin on PC12 cells after anoxia/re-oxygen in vitro
Rui ZHANG ; Hengzhao GUO ; Jidong LIU ; Hongbing CHEN ; Delin DUAN ; Yunliang GUO
Chinese Journal of Marine Drugs 2001;0(05):-
Objective To study the best protecting dosage of phycocyanin on PC12 cells after anoxia/re-oxygen in vitro.Methods The anoxia/re-oxygen cell models were established with cultured PC12 cells and treated by phycocyanin.The absorbency of PC12 cells was determined by MTT method to observe the activity and number of PC12 cells after anoxia/re-oxygen.The fluorescent intensity in PC12 cells labeled by Rhodamine123 was measured with laser scanning confocal microscope to observe the mitochondrial membrane potential after anoxia/re-oxygen.Results After anoxia/re-oxygen,the absorbency and fluorescent intensity in PC12 cells were significantly lower than those in control group.After treatment with phycocyanin,the absorbency of PC12 cell in test groups was higher than that in model group,especially in 5?g?mL-1 subgroup,P
8.Comparative study on immunoregulation of sulfated propylene glycol alginate and its fractions
Changzheng LIN ; Huashi GUAN ; Guangli YU ; Haihua LI ; Caixia GU
Chinese Journal of Marine Drugs 2001;0(05):-
Objective To investigate the influence of sulfated propylene glycol alginate(PSS) and its fractions on the immunoregulation.Methods The immunoregulation activity of PSS and its fractions were investigated by using immunocyte cultivation technique in vitro.The structure activity relationship was analysed on the basis of the structure studies of PSS' fractions.Results The experimental results showed that PSS could improve spleen cell proliferation,enhance macrophage phagocytic function and inhibit T-cell and B-cell proliferation.Conclusion PSS possessed significant immunoregulation effect,whilst the immunocompetence comparison of PSS' fractions proved that the different immunocytes had different requirements for saccharides length.
9.Immunoregulation effects of polysaccharides isolated from Octopus dollfusi on immunosuppression mice
Xiaoling LEI ; Shujin ZHAO ; Linlan JIANG
Chinese Journal of Marine Drugs 2001;0(05):-
Objective To study the immunoregulation effect of three kinds of polysaccharides extracted from different tissues of Octopus dollfusi on immunosuppression mice.Methods Two kinds of immunosuppression mice models were built by ip,cyclophosphamide(CY) and sc.hydrocortisone(HC),respectively.The spleen and thymus were weighed and the index of the immune organs was calculated.The white blood cells were counted by automated hematologic analyzer.Results In certain concentrations(20~80 mg?kg-1),the three kinds of polysaccharides could antagonize the atrophy of spleen and thymus caused by CY and HC,and increase the weight of spleen and thymus;The polysaccharides could protect against leukocytopenia induced by CY and HC,and especially in low and middle dose.Conclusion The results suggested that three kinds of polysaccharides extracted from Octopus dollfusi have potent enhancement effect on non-specific immunity for the immunosuppression mice.
10.Isolation of algicolous fungi and screening of their antitumor activity
Pengfei HAO ; Tianjiao ZHU ; Tiezheng JIA ; Qianqun GU ; Weiming ZHU
Chinese Journal of Marine Drugs 2001;0(05):-
Objective To isolate algicolous fungi from algae and obtain antitumor strains.Methods Selective isolation of algicolous fungi from algae was done,and antitumor activity corresponds to their lethiferous effect on brine shrimp and cytotoxicity against K562 cell was determined.Results 221 strains of algicolous fungi were isolated from 28 algae samples collected from intertidal zones of the Yellow Sea.Eight strains showed lethiferous activity and eight showed cytotoxicity.Conclusion The results indicated that algicolous fungi would be one of the important resources to search for new antitumor compounds.