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.

利用MSAP技术分析成体刺参的呼吸树、肠、肌肉和体壁等组织的基因组DNA甲基化水平,获得了四个组织基因组甲基化率。甲基化水平由高到低依次是呼吸树、体壁、肌肉和肠,分别是35.77%、33.51%、32.72%和28.0%,其中全甲基化位点各占19.46%、18.39%、19.18%和15.97%。统计学分析结果显示肠组织的甲基化水平与其它组织的差异显著（P＜0.05）,其余三个组织间差异不显著（P＞0.05）,但呼吸树与肌肉的半甲基化水平差异显著（P＜0.05）。由MSAP分析差异位点克隆得到四个甲基化特异性片段,经测序分析功能未知,推测为刺参基因足非编码区或新功能基因序列。

选取8个高多态性的微卫星位点,对240只刺参(Apostichopus japonicus)幼体和14只疑似亲本进行了亲缘关系的鉴定.通过检测,8个微卫星位点共得到53个等位基因,平均等位基因数为6.63;观察杂合度在0.730-0.902;各位点的多态信息含量为0.696-0.836.通过亲缘关系分析软件CERVUS 3.0分析,微卫星位点累积排除概率分别为0.9934、0.9997、0.99999.基于排除法个体基因型判定,有37个子代有1个候选亲本,194个子代有2个及以上候选亲本;基于似然法判定,置信度为80％时,有198个子代个体找到最似亲本.综合排除法和似然法的亲权鉴定,成功为229个子代个体找到它们的亲本,鉴定成功率达95％以上.本研究结果可为刺参育种工作中亲本识别及家系分析提供技术支持和理论依据.

This paper is to introduce a new paste functional cosmetic and a percutaneous absorption acne patch with competence of essential oil from wormwood and artemisia apiacea,their physicochemical index,antimicrobial effect,pox-eliminating effect have been tested,and the results are as follow,these two have good stability,compatibility,anti-bacterial activity,and certain pox-eliminating effect.

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 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 study, we analyzed a water-soluble polysaccharide MP-I isolated from Mytilus coruscus. MP-I was obtained by hot-water extraction, anion-exchange and gel-permeation chromatography. Complete hydrolysis, periodate oxidation, methylation analysis, as well as Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR) spectroscopy were conducted to elucidate its structure. MP-I was subjected to investigate the protective effect on carbon tetrachloride (CCl(4)) induced liver damage in male Kunming mice. Based on the data obtained, MP-I was found to be an alpha-(1-->4)-D-glucan, branched with a single alpha-D-glucose at the C-6 position every eight residue, on average, along the main chain. Based on the calibration with Dextran, the glucan had a molecular weight of about 1.35 x 10(6) Da. Pharmacological studies revealed that MP-I could decrease serum alanine aminotransferase (ALT), serum aspartate aminotransferase (AST), and hepatic malondialdehyde aldehydes (MDA) levels, increase the hepatic total superoxide dismutase (T-SOD) activity, and improve hepatic damage in the CCl(4) induced liver injury in mice in a dose-dependent manner. The results suggest that the possible mechanism is due to its antioxidant activity of MP-I.

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 To separate and purify polysaccharide(MP-Ⅰ) from Mytilus coruscus,and study the protective effect on CCl4-induced acute liver injury in mice.Methods The polysaccharide of MP-Ⅰ was purified using DEAE-Sepharose and Sepharose CL-6B column chromatography.The mice were treated with MP-Ⅰ via intraperitoneal injection for 7 days before CCl4 treatment.The activities of serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST),hepatic superoxide dismutase(SOD) and malondialdehyde(MDA),liver index,as well as the pathological changes of the liver were determined in all groups.Results We got pure polysaccharide MP-Ⅰ.The high(100mg·kg-1·d-1)and middle(50mg·kg-1·d-1)doses could significantly lower serum ALT and AST,hepatic SOD and MDA,and improve the liver histology.But it had no influence on the liver index of mice.Conclusion The polysaccharide MP-Ⅰ from Mytilus coruscus can protect the liver against damage induced by CCl4 in mice.

Graduate experiment is an important part of theoretical Molecular biology teaching.In this paper,some experiences were dicussed,such as how to prepare experimentation,how to choose ex- periment of better contents of experiment and how to improve students ability to fulfill experiments.

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.

BACKGROUNDAIM: To identify genes involved in cadmium-induced testicular toxicity. MATERIAL AND METHODS: We used cDNA microarray and real-time quantitative polymerase chain reaction(real-time PCR)technique to analyze the genes expression of testes in rats treated with low cadmium(4μmol/kg). RESULTS: Our studies for the first time demonstrated that expression of T-kininogen,calmegin,UDP-glucuronyl transferase,heme oxygenase and mismatch repair protein gene were associated with cadmium toxicity.However,as the organelles that produced energy,mitochondrion was not affected in the respiratory chain at low dose cadmium.In addition,vitamin C at the concentration of400mg/kg obviously attenuated cadmium-induced toxicity in testes of rats. CONCLUSION: The cadmium-reduced toxicity carcinogenic effects involved energy metabolism,defense mechanisms and DNA repair of the cell.Vitamin C might play essential roles in protecting testes of rats exposed to cadmium.

OBJECTIVE To study the effect of crude polysaccharides which were extracted from Rapana venosa,Septifer bilocularis,Turbo cornutus,Natica fortunei and Haliplanella luciae on the transformation rate of mouse lymphocytes and the multiplication rate of rat stertoli cells in vitro.METHODS The transformation rate of mouse lymphocytes and the multiplication rate of rat stertoli cells in vitro were tested with MTT method after the cells were cultured with 100 and 1 000 mg·L~(-1)polysaccharides from five kinds of marine animals respectively.RESULTS The results showed that the polysaccharides from Turbo cornutus(P0.05),Natica fortunei(low concentration group P0.05;high concentration group P0.001),Haliplanella luciae(P0.001) apparently enhanced the transformation rate of mouse lymphocytes and the multiplication rate of rat stertoli cells in vitro.CONCLUSION The crude polysaccharides from Natica fortunei,Haliplanella luciae and Turbo cornutus can enhance the transformation rate of mouse lymphocytes and the multiplication rate of rat stertoli cells in vitro.

Objective:To separate and purify polysaccharide MP-Ⅰ from Mytilus coruscus and to investigate its in vitro anti-tumor activity.Methods: The crude polysaccharide was extracted from Mytilus coruscus with hot water and the protein was removed using Sevage method.The polysaccharide MP-Ⅰ was purified using DEAE-Sepharose and Sepharose CL-6B column chromatography.Monosaccharides analysis was carried out using thin-layer chromatography and gas chromatography.The in vitro anti-tumor activity of MP-Ⅰwas assessed using MTT method.Results: The yield of MP-Ⅰwas 2.14%.MP-Ⅰ is mainly constituted by glucose.MP-Ⅰ(at 0.5,0.1,0.02 mg/ml) had different degrees of inhibitory effects on HO-8910,MCF-7, K562,and SMMC-7721 tumor cells in vitro(P0.01).The inhibitory rates of MP-Ⅰat 0.5 mg/ml against HO-8910 and MCF-7 cells were 30.55% and 36.38%,respectively.Conclusion: Polysaccharide MP-Ⅰ is mainly constituted by glucose and it has inhibitory effect on tumor cell lines HO-8910,MCF-7,K562,and SMMC-7721 in vitro.

多聚唾液酸(PSA)是一种在神经细胞黏附分子(neural cell adhesion molecule,NCAM)上表达的唾液酸聚合物,在神经发育过程中起重要作用。PSA的聚合程度会影响PSA-NCAM的功能。多聚唾液酸酶主要用于合成PSA-NCAM,两种高度同源的多聚唾液酸转移酶ST8SiaⅡ和ST8SiaⅣ都属于唾液酸转移酶家族。多聚唾液酸转移酶中NCAM的识别域和多聚唾液酸化域是截然不同的,且一些异构酶在NCAM多聚唾液酸化中起明显的负作用。多聚唾液酸酶与很多疾病都有关系,以多聚唾液酸转移酶为标靶设计的药物也将成为神经系统及肿瘤治疗的新型药物。