This work provided polysaccharide from Porphyra yezoensis Ueda for structural identification by biotechnological methods. Firstly, principal polysaccharide component was separated from P. yezoensis by water extraction and alcohol precipitation followed by chromatography. The component was identified as homogeneous polysaccharide by high performance liquid chromatography, optical rotation and ultraviolet absorption spectrophotometry. Then, structural identification was conducted by chemical methods, including periodate oxidation, reductive hydrolysis, gelatin nephelometry, methylation and desulfating analysis, and spectroscopic methods. Analysis showed that the polysaccharide component was composed of galactose (92%), as well as 3,6-inner ether galactose (3%) and 6-methyl galactose (5%) with substituents. The molecular weight was 246 kDa, and acidophobe percentage composition of 9%. It can be inferred that the polysaccharide wwas composed of precursor [(1 -> 3)-beta-D-galactose-(1 -> 4)-6-OSO3-alpha-L-galactose] and repetitive units of agarobiose [(1 -> 3)-beta-D-galactose-(1 -> 4)-3,6-inner ether-2-OCH3-alpha-L-galactose] and [(1 -> 3)-beta-D-galactose(1 -> 4)-alpha-L-galactose] in appropriate ratio of 9 : 9 : 1. Moreover, there were natural methoxy groups on the 2nd position of galactose residue with 1, 4 connection and the 6th position of galactose residue with 1, 3 connection.

This work provided polysaccharide from Porphyra yezoensis Ueda for structural identification by biotechnological methods. Firstly, principal polysaccharide component was separated from P. yezoensis by water extraction and alcohol precipitation followed by chromatography. The component was identified as homogeneous polysaccharide by high performance liquid chromatography, optical rotation and ultraviolet absorption spectrophotometry. Then, structural identification was conducted by chemical methods, including periodate oxidation, reductive hydrolysis, gelatin nephelometry, methylation and desulfating analysis, and spectroscopic methods. Analysis showed that the polysaccharide component was composed of galactose (92%), as well as 3,6-inner ether galactose (3%) and 6-methyl galactose (5%) with substituents. The molecular weight was 246 kDa, and acidophobe percentage composition of 9%. It can be inferred that the polysaccharide was composed of precursor [(1→3)-β-D-galactose-(1→4)-6-OSO3-α-L-galactose] and repetitive units of agarobiose [(1→3)-β-D-galactose-(1→4)-3,6-inner ether-2-OCH3-α-L-galactose] and [(1→3)-β-D-galactose(1→4)-α-L-galactose] in appropriate ratio of 9 : 9 : 1. Moreover, there were natural methoxy groups on the 2 position of galactose residue with 1, 4 connection and the 6 position of galactose residue with 1, 3 connection. Introduction Polysaccharide is a macromolecule active substance widely existing in flora and fauna. At present, the ocean functional polysaccharide has become a new research hotspot. Porphyra yezoensis Ueda (Fam.: Bangiaceae) is mainly distributed in Liaodong Peninsula to coastal Fujian, China. Polysaccharide in P. yezoensis contains sulphate (Yoshizawa et al. 1995) which has functions of enhancing body immunity (Yoshizawa et al. 1995), antioxidation (Isaka et al. 2015), reducing blood fat (Qian et al. 2014), antineoplastic (Yu et al. 2015a), anti-inflammatory (Isaka et al. 2015), anti-radiation and anti-aging activities. Moreover, P. yezoensis can inhibit myofibrillar protein degeneration (Jiang 2014), reduce liver injury induced by carbon tetrachloride (Guo et al. 2007), adjust intestinal flora (Kawadu et al. 1995), promote lipid metabolism (Tsuge et al. 2004) and glucose metabolism in diabetics (Kitano et al. 2012). It also can be used as genetic vector of nanoparticles of positive ion (Yu et al. 2015b). In this work, the principal polysaccharide component was separated from P. yezoensis by water extraction and alcohol precipitation to identify its primary structure. Materials and Methods Porphyra yezoensis collected from Lvsi Marine Area, Jiangsu Province, China, was dried and triturated after washing with purified water. After 3 hrs of backflow in alcohol, P. yezoensis was dried in shade. Then, water was added in solid-liquid ratio of 1 : 50 (g/m), and filtration was conducted after heat at 100oC for 3 hrs. After being concentrated, the filtrate was centrifuged at . *Co-first author, **Author for correspondence: . National Demonstration Center for Experimental Fisheries Science Education (Shanghai Ocean University), Shanghai, 201306, China. Marine Biomedicine Institute, Second Military Medical University, Shanghai, 200433, China.

In this paper, phycocyanin and one component (PY-D2) in polysaccharide were obtained from Porphyra yezoensis to study for their potential anti-tumor effects. MTT proliferation assays showed that, at concentration of 500 mg/L for 72 h. PY-D2 treatment significantly inhibited the growth of four tumor cell lines, HO-8910, MCF-7, K562 and SMMC-7721, with the respective inhibition rates of 21.2%, 23.6%, 19.8%, and 21%. Flow cytometry analysis indicated that the anti-tumor effect of PY-D2 was associated with the cell cycle arrest at G0/G1 or G2/M check-points. Two cancer cell lines, Hep-2 and A375 were treated by different concentrations of phycocyanin, followed by radiation with He-Ne laser at the density of 20 J/cm^2 and the wavelength of 632 nm. MTT assays displayed the minimal survival rate of 29.8% (Hep-2) and 16.2% (A375) at 100 μg/ml and 165 μg/ml respectively. These results suggest that both polysaccharide and phycocyanin from Porphyra yezoensis might be useful in the treatment of human cancers. These results suggest that phycocyanin and one component (PY-D2) in polysaccharide which had been obtained both from Porphyra yezoensis were potential medicaments in the treatment of some kinds of human cancers.

The reaction between ErCl3·6H2O and H2bmdc (H2bmdc = benzimidazole-5,6-dicarboxylic acid) produced two distinct complexes: 1D helical complex {[Er(Hbmdc)(bmdc)(H2O)3]·3H2O}n (1) obtained under hydrothermal condition at 160°C and discrete complex [Er2(Hbmdc)2(bmdc)2(H2O)8]·8H2O (2) under ambient temperature, evaporated from the mother liquor of 1. The crystal structures of the complexes were determined by X-ray single-crystal diffraction. Through comparison of complexes 1 and 2, we found that enhanced temperature favors the formation of higher dimensional products and lower amount of coordinated water. Both complexes are characterized by IR, elemental analysis, X-ray powder diffraction and thermogravimetric analysis.

The ex vivo antioxidation activity of polysaccharide extracted from the red seaweed Porphyra vezoensis was studied by systematically measuring and analyzing the scavenging efficiencies on the free radicals O- 2·, OH and DPPH, and the inhibitory effects on the hemolysis in mouse erythrocytes induced by H2O2 and on lipid peroxidation in mouse liver homogenates. The results show that Porphyra polysaccharide, in the experimental concentration range, possessed antioxidation activity. The scavenging efficiency for O- 2· was found to be remarkably high and the maximum scavenging rate was 82.77%. The scavenging efficiency for -OH was even higher than for O- 2·, with a maximum removing rate of 85.63%, whereas for DPPH it was 13.97%. The inhibitory effects on mouse erythrocyte hemolysis and malondialdehyde formation in mouse liver were significant, with maximum inhibition rates of 82.90% and 58.48%, respectively. The combined data indicate that the polysaccharide extracted from Porphyra has strong antioxidation activity.

In this paper,high purified R-Phycoerythrin(R-PE)and C-Phycocyanin(C-PC)were extracted from Porphyra yezoensis,and they were applied as photosensitizer to study on inhibitory effect of photodynamic reaction on human laryngeal cancer cell line Hep-2.With MTT assay,effects of R-Phycoerythrin and C-Phycocyanin concentrations(10,25,50 and 100μg/ml)and illumination dose on cancer cell photodynamic therapy reaction with irradiation of iodine tungsten lamp were analyzed.The results showed that the photodynamic effects of these two phycobiliproteins on Hep-2 were remarkable.With 100μg/ml concentration of C-PC and R-PE and 50J/cm2 illumination dose of iodine tungsten lamp,the survival rates of the cancer cells were 64% and 57%,respectively.The survival rates of the cancer cells only treated by illumination of iodine tungsten lamp was 86.9%.The inhibition rates of the cancer cells treated only with higher concentration of phycobiliproteins(R-PE,C-PC≥50μg/ml)were about 68% after they cultured for 24hs.It was indicated that the R-Phycoerythrin and C-Phycocyanin extracted from Porphyra yezoensis would become a new kind of photosensitizer and could be applied to photodynamic therapy of cancer.

Objective:To observe the anti-tumor effects of Porphyra yezoensis polysaccharide.Methods:By extracting and purifing of polysaccharide from P.yezoensis,two components,PY-D1 and PY-D2,were isolated with ion-exchange chromatography DEAE-52.In vitro,the growth of four different human tumor cell lines were detected by MTT and flow cytometric analysis after induced by P.yezoensis polysaccharide PY-D2 for 72 h.Results:PY-D2 could significantly inhibit the growth of human tumor cells HO-8910,MCF-7,K562 and 7721.The inhibitory rates were 21.2%,23.6%,19.8% and 21% separately after cells were induced by 500 mg/L PY-D2 induced(P0.001).The result of flow cytometry indicated that PY-D2 showed the antitumor effect of PY-D2 was relative to the blocking of G0/G1 or G2/M period cells.Conclusion:P.yezoensis polysaccharide PY-D2 has showed cytotoxicity effect on the tumor cells and may be a potential adjuvant to cancer therapy.

目的:研究条斑紫菜多糖体外提高机体免疫力的作用。方法:应用生化技术分离和纯化条斑紫菜多糖,获得条斑紫菜多糖2个组分,分别为PY-D1和PY-D2。体外培养条件下分别用不同浓度的PY-D2以及PY-D2与ConA或LPS协同处理小鼠脾淋巴细胞,通过MTT法观察条斑紫菜多糖对小鼠脾淋巴细胞生长的影响。采用流式细胞仪检测小鼠脾淋巴细胞的细胞周期变化。结果:PY-D2处理小鼠脾淋巴细胞72小时后对其生长有明显促进作用,且呈剂量依赖效应,0.25,0.5和1mg/ml条斑紫菜多糖处理小鼠脾淋巴细胞后,小鼠脾淋巴细胞的存活率分别为157.5%,162.1%和173.4%（P＜0.01）。PY-D2与ConA或LPS共同处理小鼠脾淋巴细胞时,小鼠脾淋巴细胞的存活率高于ConA和LPS单独的作用,表现出明显的协同效应。流式细胞仪检测表明PY-D2可以促进小鼠脾淋巴细胞从G1期进入S期。结论:PY-D2可以促进小鼠脾淋巴细胞生长,为今后研究多糖提高机体免疫力的作用机理奠定了坚实的基础。

In this paper,anti-fatigue effects of Porphyra yezoensis polysaccharide and their dose-effect relationship was studied with live-animal experiment.By measureing the effect of different dosage of Porphyra polysacccharide on mice swimming time,lactate dehydrogenase(LDH)activity of mice before and after swimming,and the content of muscle glycogen and hepatic glycogen of mice,for identifying Porphyra polysaccharide could increase anti-fatigue bioactivity of mices.The experiment results showed that PY polysaccharide could prolong mice swimming time signifcantly,increasing as high as 37%,and it could enhance the reserves amount of muscle and hepatic glycogen,by 185.6% and 186% respectively;and the polysaccharide also could increase LDH activity by 135% and 137.5% before and after swimming respectively.It indicated that polysaccharide from Porphyra yezoensis had the antifatigue bioactivity.

To study the protective effect and possible mechanism of Porphyra yezoensis polysaccharide (PYP) in hepatotoxicity mice, acute liver injury was successfully induced by injecting 0.2% carbon tetrachloride (CCl4) intraperitoneally. Levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum and liver homogenate, content of malondialdehyde (MDA), activities of total superoxide dismutase (T-SOD) in liver were measured by biochemical methods. Liver index was calculated and pathological changes of the liver tissue were observed microscopically. PYP was found to significantly decrease the activities of ALT and AST (P < 0.05), to remarkably lower the liver indexes and MDA level in hepatical tissues in mice (P < 0.05), and to upregulated the lower T-SOD level in liver homogenate (P < 0.01). Furthermore, histologic examination showed that PYP could attenuate and the extent of necrosis, reduce the immigration of inflammatory cells. PYP plays a protective action against hepatotoxicity induced by CCl4 in mice, and its mechanisms may be related to free radical scavenging, increasing SOD activities and anti-lipid peroxide.

The polysaccharide from Porphyra yezoensis(PYP) was purified and analyzed by biochemical techniques. The effect of PEP on the proliferation of MCF-7 was investigated in vitro. Methods: The purified polysaccharide from Porphyra yezoensis was obtained by DEAE-cellulose and Sephadex G-200 column chromatography. The effect of PEP on the proliferation of MCF-7 was investigated in vitro by MTT method. Two components, PY-D1 and PY-D2, were isolated from the Porphyra yezoensis polysaccharide by ion-exchange chromatography DEAE-52. Two polysaccharides, PY-G1 and PY-G2, were further isolated from PY-D2 Ueda by gel filtration Sephadex G-200. PY-G was identified by UV and IR spectra. All of components could inhibit the proliferation of MCF-7. Results: It was found polysaccharides of Porphyra yezoensis had antitumor activity for MCF-7 cells.

Objective:In order to research some biological functions of crude polysaccharide from Porphyra yezoensis,the lymphocytes and the sertoli cells were deteced.Methods:After primary culturing,the spleen lymphocytes of mice and rats and the sertoli cells were separated into 96-well cell culture plates.Different concentrations of crude polysaccharide from P.yezoensis(0.05,0.5,5,10 mg/mL)were selected and added to the culture.The cell survival rates were assessed by MTT assay.Results:Obviously,the crude polysaccharide from P.yezoensis can enhance the proliferation of these cells.When the concentration of the polysaccharide is 10 mg/mL,the rate of the proliferation reached 137.3%,149% and 454.5% respectively.Conclusion:The results indicates that crude polysaccharide from P.yezoensis may improve the immunity and the procreative function.Especially,the effect on the proliferation of the sertoli cells has not been reported yet.

Ti tartrate complex (Ti(OiPr)4+DET)-grafted hexagonal mesoporous silica (HMS) catalysts have been successfully prepared through three assembled pathways, and show the high catalytic activity and a definite epoxidative enantioselectivity for the epoxidation of styrene with tetrabutyl hydroperoxide (TBHP) as an oxidant.

Three types of HMS supports, which were as-synthesized, HMS(u), HMS(c) calcined from HMS(u), and HMS(m) prepared by the modification of HMS(c) with 3-amino propyltriethoxysilane, were first used to assemble the Ti catalytic sites through the exchange reactions between them and Ti compounds such as Ti tetraisoproxide (Ti(OiPr)4), Ti tetrachloride (TiCl4)) and their Ti tartrate complexes. The Ti-assembled HMS catalysts were investigated in detail by means of FT-IR, UV–vis reflection spectra, chemical and elemental analyses. It is found that in Ti(OiPr4 and especially TiCl4, the exchange that takes place with the surface hydroxyls of the HMS(c) is easier than with the surface NH2 groups of HMS(u) and HMS(m). Ti tartrate complex can be successfully assembled on the HMS(c) through three assembled pathways. The UV–vis reflection spectra indicates that the Ti species are highly dispersed upon the surface of HMS. In catalyzing epoxidation of styrene with TBHP, the above-described catalysts show a higher catalytic activity and selectivity for the epoxides than unloading Ti tartrate complex, and the Ti-substituted HMS catalysts. Their outstanding advantage is that they can be reused many times with little loss of activity.

Pure silicon and Ti-substituted mesoporous molecular sieves(HMS and MCM-41) have been synthesized by surfactant templpating pathway.A type of Ti(O iPr) 4+DET chiral complex has been successfully assembled to mesoporous carriers by use of exchange reaction of the surface hydroxyls of mesoporous carriers with Ti compound or d-diethyl tartrate(DET).Though four assembled pathways,chiral Ti anchored mesoporous catalysts have been first prepared.These anchored chiral catalysts are characterized by FT-IR.Its Ti contents are measured.In catalyzing the epoxidation of styrene with tertbutyl hydroperoxide(TBHP) as an oxidant,these chiral catalysts have the better catalytic activity and the selectivity for the epoxides.The adjusting interaction of DET ligand to the Ti sites anchored on the mesoporous carriers will cause the decrease in the catalytic activity and the increase in the epoxidation selectivity of the Ti sites.The chiral inducting interaction of DET will induce to produce the enantiomeric excess of d-epoxide(ee.% is up to 32.4%)

Mesoporous silicas MCM 41 and HMS were synthesized by the cooperative assembly of silica and surfactants. Two methods are raised to assemble titanium tartrate complexes over the untreated and calcined mesoporous carriers. The results of XRD and FT IR indicate that MCM 41's mesostructure with long range order will deteriorate, and be transformed into the mesostructure similar to HMS. This deterioration of the mesostructure arises the disappearance of 1?219 and 580?cm -1 , and then also causes a stronger aberrance of MCM 41's Si-OH groups. The binding of HMS with Ti complexes will induce the Si-OH groups of HMS to produce a weaker aberrance. In the epoxidation of styrene, the assembled Ti catalysts with the calcined mesoporous carriers exhibit the highest oxidative activity, and the assembled Ti catalysts with untreated mesoporous carriers possess the best epoxide selectivity. The modification of the templating agents and diethyl tartrate (DET) ligand on the Ti active centers will cause the decreases in the actalytic activity and the increases in the selectivity.

investigated the synthesis conditions of n-pro pyl propionate catalyzed with ammonium ferric sulfate dodecahydrate,reported the catalytic syntheses of n-propyl formiate, n-propyl acetate, n-propyl propionate, n-propyl butyrate and n-propyl isovalerate using ammonium ferric sulfate dodecahydrate.

XRD and IR show that titanium is effectively embedded into framework of HMS, and the order degree of mesoporous HMS doesn't fall down.Embedding content of Ti corresponds well with n(Ti)/n(Si), and the most one is up to 1.61 mmol Ti per g HMS. The intensity of 960 cm -1 band on the IR spectra of Ti-HMS samples takes direct proportion to embedding Ti content. It is shown that this peak may be caused with distortion of SiO 4 tetrahedron that titanium atom incorporate into the mesoporous framework. In catalyzing the epoxidation of styrene, Ti-HMS catalyst can effectively catalyze the oxidation of styrene by TBHP into styrene oxides (65%～86%), phenylacetaldehyde (6%～15%) and benzaldehyde (8%～20%). Be used repeatedly without to be loss of activity. The preparation and reaction conditions of Ti-HMS have the different influences on its catalytic activity and product selectivity.

Cobalt-containing hexagonal mesoporous molecular silica(Co-HMS)was prepared by using neutral dodecylamine (DDA)as templating agent at room temperature,and characterized by means of XRD,FT-IR,SEM,etc..XRD and FT-IR measurements indicate that the incorporation of Co obviously decreases the assembled effects of templating agent,and deteriorate the quality of the 100 reflection of mesoporous materials and the frame of Si-O.The catalytic tests shows Co-HMS is a kind of effectice catalyst for hydroxylation of phenol with 30% H 2O 2 in aqueous solution,but have no catalytic activity in the reactions of epoxidation of styrene and cyclohexene.