微丝解聚剂及微管阻断剂对藓羽藻细胞重建过程的影响

某些种类的海洋多核绿藻的原生质被挤出后,其细胞器在人工海水中可立即发生聚集,随后其细胞组分发生重建,形成新的细胞膜与细胞壁,这些重建的细胞在最佳培养条件下,最多有40%能够发育成成熟藻体。细胞骨架系统是细胞内有序性的维系者,也是细胞及细胞器运动的动力源,这些过程都涉及细胞骨架的解聚与重聚合的动态过程。本文利用微丝解聚剂细胞松弛素B及微管阻断剂秋水仙素对藓羽藻(Bryopsis hypnoides)细胞重建过程的影响进行了研究。结果显示,细胞器聚集及聚集体变圆的动力不来自于细胞骨架系统,而原始微丝体系的解聚是细胞器高效聚集的基本条件;细胞松弛素B及秋水仙素都显著抑制细胞膜的形成及细胞内组分有序性重建等后继发育过程,表明细胞骨架系统在这些过程中扮演着重要角色。     

Effect of Fe3+ on the growth and lipid content of Isochrysis galbana

Inducing lipid accumulation in microalgae cells without suppressing cell growth is vital to the economical production of biodiesel from microalgae. In two experiments, we demonstrate that the cell concentration and lipid content of marine microalgae Isochrysis galbana depend upon the iron concentration in the growth media. In Experiment I, adding chelated FeCl 3 to the medium at the late exponential growth phase prolonged this phase and increased the lipid content in I. galbana cells. The fi nal cell density and lipid content of I. galbana supplemented with chelated FeCl 3 was approximately 2 and 1.65 times higher than that of non-supplemented cultures, respectively. In Experiment II, I. galbana cells in the late exponential phase were collected and re-inoculated into new media containing Fe 3+ at various concentrations. The fi nal cell concentration and lipid content were maximized at the highest iron concentration(38% biomass by dry weight at 1.2×10-5 mol/L Fe 3+). In this study, intracellular neutral lipid storage was evaluated by fl uorescent spectrophotometry using fl uorochrome Nile red, and the measurement conditions were optimized.     

Ultrastructure Developments During Spermiogenesis in Polydora ciliata （Annelida： Spionidae）,a Parasite of Mollusca

Spionid worms of Polydora ciliata inhabit the shells of many commercially important bivalves and cause disease in molluscan aquaculture. Their sperm structure is closely related to their fertilization method. To give an insight into the sperm structure and spermatogenesis, ultrastructure details of the subcellular components of germ cells during spermiogenesis of Polydora ciliata are detected by transmission electron microscopy （TEM）. In P. ciliata, during spermiogenesis, chromatin is regularly arranged as dense fibrils and becomes more condensed when the nucleus elongates. Microtubules do not surround the nucleus during its elon- gation. The Golgi phase is characterized by the formation of proacrosomal granules within the Golgi apparatus. The proacrosomal granules fuse to form a single, spherical acrosomal vesicle that migrates to the anterior pole of the cell. At the time of nuclear con- densation, mitochondria become reduced in number but increased in size, causing deep indentation at the base of the nucleus. The mid-piece has a few mitochondria. The cap phase includes the spreading of the acrosomal granule over the surface of the nucleus of the differentiating spermatid. The acrosomal phase of spermiogenesis is typically associated with changes in the shape of the nucleus, acrosome and tail. The relationship of sperm ultrastructure to spermiogenesis in spionidae species was discussed.     

Purification and photodynamic bioactivity of phycoerythrin and phycocyanin from Porphyra yezoensis Ueda

Phycoerythrin and phycocyanin were purified from Porphyra yezoensis Ueda with their bioactivity determined in this study. Continuous precipitation with ammonium sulfate at different concentrations （10%, 20%, 40%and 50%） increased the purity （A564：A280） of phycoerythrin to 1.49, 3.92 fold of the raw extract （0.38） and the purity （A615：A280） of phycocyanin to 0.70, 3.33 fold of the raw extract （0.21）. Two more times of chromatography with hydroxylapatites finally made the purity of phycoerythrin and phy-cocyanin reach 5.50, 14.47 fold of the raw extract, and 5.10, 24.29 fold of the raw extract, respectviely. The yield of high purity phycoerythrin and phycocyanin were 0.21%and 0.09%of dried P. yezoensis blade, respectively. The photodynamic cytotoxic ex-periment showed that both phycoerythrin and phycocyanin inhibited the growth of liver tumor cells significantly. It was found that 250 mg L-1 purified phycoerythrin and phycocyanin inhibited the growth of hepatocellular carcinoma cells 24 h after laser-irradiation by 80%and 59%, respectively, and 100 mg L-1 purified phycoerythrin and phycocyanin induced the apoptosis of 31.54%and 32.54%of the cells, respectively, 8 h after photodynamic therapy. Oue findings demonstrated that P. yezoensis can serve as photosensitizer （phycoerythrin and phycocyanin） producer.     

PI3K/Akt pathway is involved in the activation of RAW 264.7 cells induced by hydroxypropyltrimethyl ammonium chloride chitosan

We previously demonstrated that 2-hydroxypropyltrimethyl ammonium chloride chitosan(HACC)promoted the production of nitric oxide(NO)and proinflammatory cytokines by activating the mitogen-activated protein kinases(MAPK)and Janus kinase(JAK)/STAT pathways in RAW 264.7 cells,indicating good immunomodulatory activity of HACC.In this study,to further investigate the immunomodulatory mechanisms of HACC,we determined the roles of phosphatidylinositol 3-kinase(PI3K)/Akt,activating protein(AP-1)and nuclear factor kappa B(NF-κB)in HACC-induced activation of RAW 264.7 cells by the western blotting.The results suggest that HACC promoted the phosphorylation of p85 and Akt.Furthermore,c-Jun and p65 were also increased after the treatment of RAW 264.7 cells with HACC,indicating the translocation of NF-κB and AP-1 from cytoplasm to nucleus.In addition,as scanning electron microscopy(SEM)analysis shows,the cell morphology changed after HACC treatment.These findings indicate that HACC activated MAPK,JAK/STAT,and PI3K/Akt signaling pathways dependent on AP-1 and NF-κB activation in RAW 264.7 cells,ultimately leading to the increase of NO and cytokines.     

镉诱导长江华溪蟹(Sinopotamon yangtsekiense)心肌细胞凋亡的研究

采用急性毒性实验方法,研究了不同浓度氯化镉(0、7.25、14.50、29.00、58.00和116.00mg/L)处理不同时间(1d、3d、5d和7d)对长江华溪蟹心肌细胞凋亡的影响。结果表明,低浓度组及染毒时间较短时镉对心肌的影响不明显,但随着染毒浓度的增加和时间的延长,心肌出现空泡变性、玻璃样变性、炎细胞浸润等病理现象,在染毒7d虽然观察到疑似的凋亡细胞,但凋亡率较低;荧光显微镜未观察到心肌细胞典型的凋亡细胞核形态学变化;琼脂糖凝胶电泳图谱未出现凋亡细胞所特有的梯状条带;流式细胞术检测结果显示镉处理组细胞凋亡率增加,但与对照组相比,无统计学意义(P>0.05);Caspase-3活性也没有显著性变化(P>0.05)。因此,在本实验所选取的浓度和时间范围内,镉未诱导长江华溪蟹心肌细胞发生细胞凋亡,这与本课题组前期研究中观察到的肝胰腺、精巢组织在染毒2d、7d发生细胞凋亡的情况明显不同,这可能与心脏组织的特殊性、不同组织对镉的积累量不同有关。     

Longitudinal patterns of spring-intermonsoon phytoplankton biomass, species compositions and size structure in the Bay of Bengal

Vertical distributions of phytoplankton biomass, compositions and size structure were investigated during the spring-intermonsoon (April 22 to 30) of 2010 along transact 10°N of the Bay of Bengal, northern Indian Ocean. Surface phytoplankton biomass (Chl a) was (0.065§0.009) μg/L, being greater than 80% of which was contributed by pico-phytoplankton (<3 μm). The Chl a concen- tration vertically increased to the maximal values at deep chlorophyll maximum (DCM) layer that shoaled eastwards from 75 to 40 m. The Chl a biomass at DCM layer generally varied between 0.2 and 0.4 μg/L, reaching the maximum of 0.56 μg/L with micro-phytoplankton cells (>20 μm) accounting for 58% and nano- (3-20 μm) or pico-cells for 15% and 27%, respectively. In particular, the cells concentration coupling well with phosphate level was observed at middle layer (75-100 m) of 87° to 89°E, dominated by micro-cells diatoms (e.g., Chaetoceros atlanticus v. neapolitana, Chaetoceros femur and Pseudonitzschia sp.) and cyanobacteria (i.e., Trichodesmium hildebrandtii), with the cells concentration reached as high as 4.0×10⁴ and 4.3×10⁴ cells/L. At the rest of the trans- act however, dino°agellates (e.g., Amphisdinium carterae and Prorocentrum sp.) were the dominant species, with the cells concentration varying from 0.3×10³ to 6.8×10³ cells/L. Our results also in- dicate that the regulation of large cells (micro-, nano-) on phytoplankton biomass merely occurred at DCM layer of the Bay.     

坛紫菜叶状体营养细胞与生殖细胞叶绿素荧光特性比较

利用叶绿素荧光技术对坛紫菜(Porphyra haitanensis)叶状体营养细胞和生殖细胞的光能利用特性进行了比较。结果表明:坛紫菜叶状体营养细胞和雌性生殖细胞的实际量子效率(ΔF/Fm′)差异不明显,但显著高于雄性生殖细胞;快速光曲线测定表明雌、雄藻体营养细胞的最大相对电子传递速率(rETRmax)相近,但显著高于雄性生殖细胞;不同生殖细胞半饱和光强(Ik)无显著差异;在生长光强下,营养细胞和雌性生殖细胞rETR、光化学猝灭(qP)和非光化学猝灭(NPQ)差异不明显,而雄性生殖细胞rETR、qP等荧光参数均显著低于营养细胞和雌性生殖细胞。本文表明坛紫菜叶状体营养细胞和雌性生殖细胞具有较高的光能利用能力,能够将吸收的光能多数用于电子传递,而雄性生殖细胞对光能的利用能力较低。     

海洋疣荔枝螺体外抗肿瘤活性

荔枝螺是福建海域常见食用螺类,古方记载和民间长期食用证实具有良好药用价值,但国内外对其生物活性物质的研究还较少.本研究以福建海域疣荔枝螺(Thais clavigera)为研究材料,从疣荔枝螺消化腺部位获得抗癌组分E-EA-2,通过液相色谱、气质联用色谱分析发现其主要成分为6-溴靛红(6-bromoisatine),分子结构式C_8H_4BrNO_2.E-EA-2能有效抑制人乳腺癌MCF-7、人肝癌QGY-7703细胞增殖,药物浓度高于40μg/cm~3时,癌细胞存活率显著降低.E-EA-2能诱导肝癌细胞QGY-7703发生PARP蛋白剪切,加入泛caspase抑制剂使细胞存活率显著提高.同时,E-EA-2使MCF-7细胞核产生固缩,线粒体膜电位显著下降,凋亡细胞数量显著增加.E-EA-2对肝癌细胞QGY-7703的IC50为46μg/cm~3,对乳腺癌细胞MCF-7的IC50为28μg/cm~3,对癌细胞的增殖抑制作用主要是以促凋亡方式实现的.