硒化牡蛎多糖制备及其抗氧化和抗肿瘤活性研究

亚硒酸钠和牡蛎多糖合成硒化牡蛎多糖(SeOPS),纤维素DE-52分离纯化,红外光谱结构表征。DPPH·,ABTS+和·OH清除作用及铁还原力测定SeOPS抗氧化活性。MTT法检测SeOPS抑制肿瘤细胞增殖能力,采用流式细胞术检测SeOPS对细胞周期和凋亡的影响。结果表明,SeOPS具有清除DPPH·和ABTS+作用,IC50值分别为7.102 mg/m L和2.243 mg/m L。·OH及铁还原力清除效果与牡蛎多糖(OPS)无显著差别。SeOPS阻滞人宫颈癌(Hela)细胞G0/G1期及肝癌(HepG2) S期的发育,诱导细胞凋亡,从而抑制肿瘤细胞的增殖。     

表皮生长因子对牛输卵管上皮细胞生长、增殖和凋亡的影响

研究不同浓度的表皮生长因子对牛输卵管上皮细胞生长、增殖和凋亡的影响,目的是建立高质量的牛体外胚胎共培养细胞系。牛输卵管上皮细胞体外培养技术结合血球计数法检测细胞增殖,并利用流式细胞仪进行细胞周期和凋亡的检测,通过Lysis软件对数据进行分析。结果表明,EGF在0～50 ng/mL浓度范围内,其促增殖、抑制凋亡作用随浓度的增加而增大,呈剂量依赖性;当浓度持续增高时,其促增殖、抑制凋亡能力降低;此外,S期细胞数量明显增加,G2-M期细胞数量有下降趋势。表皮生长因子在一定浓度范围内具有促进牛输卵管上皮细胞增殖和抑制其凋亡的作用,并使其细胞周期发生变化。     

硫脲对水稻幼苗生长和部分生理指标的影响

充分吸胀的水稻种子经0￣1000mg/L的硫脲溶液培养,随着硫脲浓度增加,幼苗生长受到抑制,表现在株高、根长、根数、干重明显降低,叶绿素、类胡萝卜素含量及膜脂过氧化产物丙二醛(MDA)含量下降;硫脲浓度50￣500mg/L使幼苗可溶性糖含量增加、500mg/L以上浓度使可溶性蛋白含量降低、300mg/L以上浓度使SOD活性下降。     

氯化钴处理微藻的生物学效应初报

在塔胞藻培养液中加入不同浓度的CoCl2(0、2、4、6、8、10、12mg/L),然后分析了氯化钴处理塔胞藻的生物学效应。结果发现,当培养液中钴离子浓度较低时,塔胞藻被处理后的生物学效应明显优于对照。当钴离子浓度达到一定值(6mg/L)时,则随着钴离子浓度的升高,细胞的生长繁殖在前2轮(9d为一轮)培养中均明显降低;细胞内的蛋白质、叶绿素含量也均明显下降,而可溶性糖的含量与对照相比变化不大。继续培养,塔胞藻细胞的生长繁殖及细胞内含物含量接近正常值。     

高Fe2+胁迫下水稻幼苗根尖铁的分布及细胞超微结构变化

以耐性水稻(Oryza sativa L.)品种Azucena为实验材料,研究高Fe2+胁迫下,0～10mm段幼苗根尖铁的分布,以及根尖细胞亚显微结构变化情况.结果表明,400μmol/L Fe2+处理下,0～2mm段根尖铁含量最高;随着Fe2+浓度的升高,根尖边缘细胞(BC)铁含量呈递增趋势.电镜观察发现,400μmol/L Fe2+处理下,BC内出现大量碎片,受损明显,而根冠细胞主要表现为细胞壁增厚、细胞内出现许多小液泡.     

镉胁迫对大球盖菇菌丝生长及抗氧化系统的影响

本研究以质量浓度为0、0.2、0.6、1.0、2.0和10 mg/L的Cd胁迫培养的大球盖菇(Stropharia rugosoannulata)菌丝为试验材料,研究不同浓度Cd胁迫对菌丝中Cd积累量、菌丝生长状况及形态、细胞膜氧化损伤、非酶类抗氧化剂、以及抗氧化酶活性的影响。结果表明,随着Cd胁迫浓度的升高,冻干菌丝中积累的Cd含量逐渐升高,从0.530 mg/kg升高至669.91 mg/kg,存在显著差异性(P0.05)。Cd胁迫对大球盖菇的菌丝生长具有抑制作用,对生长速度的抑制率为10%～36%。Cd胁迫浓度的增加,促进了丙二醛(MDA)含量的升高,同时也促进了非酶类抗氧化剂抗坏血酸(ASA)含量、还原型谷胱甘肽(GSH)含量的增加,使其都在10 mg/L的Cd处理浓度下达到最高值,分别为对照组的2.63、1.88和18.49倍。过氧化氢(H2O2)含量则先上升后下降,被H2O2激发的谷胱甘肽还原酶(GR)、谷胱甘肽过氧化物酶(GPX)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)和超氧化物歧化酶(SOD)活力也都呈现先上升后下降的趋势。低浓度Cd胁迫时,大球盖菇菌丝可能主要靠SOD和CAT来清除细胞中的活性氧(ROS);高浓度Cd胁迫时,则可能主要靠抗坏血酸-谷胱甘肽循环(As A-GSH)来清除细胞中的ROS。本试验为深入开展大球盖菇菌丝应答Cd胁迫的分子机制研究提供科学依据。     

氯化钻处理微藻的生物学效应初报

在塔胞藻培养液中加入不同浓度的CoCl2（0、2、4、6、8、10、12mg／L），然后分析了氯化钴处理塔胞藻的生物学效应。结果发现，当培养液中钴离子浓度较低时，塔胞藻被处理后的生物学效应明显优于对照。当钴离子浓度达到一定值（6mg/L）时，则随着钴离子浓度的升高，细胞的生长繁殖在前2轮（9d为一轮）培养中均明显降低；细胞内的蛋白质、叶绿素含量也均明显下降，而可溶性糖的含量与对照相比变化不大．继续培养．塔胞藻细胞的生长繁殖及细胞内含物含量接近正常值。     

玉米种子萌发期根缘细胞对 NaCl 胁迫响应

NaCl胁迫是导致玉米盐害的主要因素,研究了NaCl对玉米萌发期根系和离体根缘细胞的影响。结果表明,随着NaCl浓度从0 mmol/L增加到250 mmol/L,胚根长和次生根数量受到显著抑制,同时根缘细胞存活率也显著降低。但在100 mmol/L的NaCl胁迫下,根缘细胞的数量比对照和高浓度胁迫有显著增加,同时100 mmol/L的NaCl胁迫也显著刺激了玉米离体根缘细胞的凋亡速度。由于试验材料对100 mmol/L的NaCl胁迫有一定耐性,因此,认为该胁迫浓度下根缘细胞数量增加可能与其耐盐性有关,为进一步探讨根缘细胞与玉米耐盐机制的关系奠定了基础。     

铅（Pb）对西葫芦幼苗生理生化特性的影响

用不同浓度的1/2 Hoagland+Pb2+做胁迫试验,研究重金属铅(Pb)对西葫芦生理生化特性的影响.结果表明,随着Pb2+浓度的升高,西葫芦幼苗叶绿素含量下降,以及叶绿素a/b比值明显上升,叶片细胞膜透性显著增加,丙二醛(MDA)含量显著升高.随着Pb2+浓度的升高,超氧化物歧化酶( SOD)活性、过氧化物酶(POD)活性、过氧化氢酶(CAT)活性逐渐升高,达到一定浓度时,SOD、POD、CAT活性则显著下降.     

CO2浓度升高对春小麦不同生育时期抗氧化系统的影响

利用开顶式气室研究了CO2浓度升高(550μmol/mol)对小麦(Triticum aestivumL)孕穗期和开花期抗氧化系统的影响。结果表明,高浓度CO2下,小麦叶片外渗电导率和丙二醛的含量下降,说明膜脂过氧化程度有所降低;虽然O2产生速率和H2O2含量2个时期相比开花期大于孕穗期,但是两者均低于对照,说明高CO2浓度下活性氧产生减少;SOD,POD,CAT的活性与对照相比有所增加并达到显著或极显著水平;随着CO2浓度的升高,ASA和Car含量也随之增加,但是随着生育期的推移,两者含量均有不同程度的减少;最终生物量和籽粒产量分别增长18%和14%,说明CO2浓度升高有利于减轻小麦的氧化损伤,促进植物生长。     

Apoptosis of Hepatoma Cells HepG-2 Induced by Shikimic Acid from FRUCTUS ANISI STELLATI and Its Mechanism

[Objective]To research the apoptosis of hepatoma cells HepG-2 induced by shikimic acid from FRUCTUS ANISI STELLATI and its mechanism. [Methods]Hepatoma cells HepG-2 at logarithmic phase were randomly divided into drug group and control group. Shikimic acids from FRUCTUS ANISI STELLATI at the concentrations of 0. 125,0. 5 and 1 g /L were used. Inhibitory effects of shikimic acids on HepG-2 cell proliferation were detected by MTT method. Effects of shikimic acids at the concentrations of 0. 125,0. 5 and 1 g /L on the cell cycle of hepatoma cells HepG were researched by Flow Cytometer. Expression of Bcl-2 and Bax proteins were detected by immunocytochemistry detection. [Results]Results of MTT test showed that shikimic acid(0. 125,0. 5 and 1 g /L) had significant inhibitory effects on the proliferation of in vitro human liver cancer HepG-2 in a time-and dose-dependent relationship(P 0. 05). Shikimic acid blocked the hepatoma cells HepG-2 at G1 phase,so that the cells could not enter into S phase for DNA synthesis,which restricted the cell proliferation. Results of immunocytochemistry detection showed that protein expression of Bcl-2 decreased significantly,while Bax expression enhanced significantly,so that the proportion of Bcl-2 to Bax reduced. [Conclusions]shikimic acid blocked the hepatoma cells HepG-2 at G1 phase,and had significant inhibitory effects on the proliferation of human hepatoma cell through reducing the proportion of Bcl-2 to Bax.     

大型海藻养殖对水体DO含量的影响

为了解大型海藻养殖对海水DO含量的影响,明确其在光照、黑暗及光暗交替条件下海水DO含量的变化过程。笔者将取自温州洞头近海养殖的鼠尾藻、羊栖菜带至实验室内进行养殖实验,设置光照（2、6、12 h）、黑暗（2、6、12 h）、光暗交替（1 h 1 h、6 h 6 h、12 h 12 h）处理,测定养殖水体DO含量,对其产氧速率、耗氧速率等进行分析。结果表明：随着光照时间的延长,鼠尾藻、羊栖菜产氧速率都逐渐下降,鼠尾藻的产氧速率比羊栖菜高（P<0.05）;黑暗条件下,鼠尾藻耗氧速率在6 h时达最高288.02 mg/kg DW?h,羊栖菜耗氧速率在2 h时达最高207.46 mg/kg DW?h,鼠尾藻的耗氧速率比羊栖菜高;光暗交替下鼠尾藻的净产氧速率先急剧降低后缓慢升高,短光暗交替周期（1 h 1 h）时净产氧速率最高457.08 mg/kg DW?h。羊栖菜净产氧速率先升高后降低,在中光暗交替周期（6 h 6 h）时最高64.04 mg/kg DW?h。大型海藻养殖时随着光暗环境的交替进行,不断发生着光合产氧、暗呼吸耗氧的转换,总体而言海藻养殖提高了水体DO含量。大型海藻养殖的生态、社会价值显著,建议大力推广     

Effects of Water Decoction of Plumbago zeylanica L. on the Proliferation,Apoptosis and Cell Cycle of Rats Hepatic Stellate Cells

[Objective] To observe the effects of water decoction of Plumbago zeylanica Linn.on the proliferation,apoptosis and cell cycle of rats hepatic stellate cells,and to discuss the function and mechanism of anti-hepatic fibrosis of P.zeylanica.[Methods] SD rats were given water decoction of P.zeylanica by gavage,so as to obtain medicated serum.Medicated serum was incubated together with hepatic stellate cells( HSC-T6) according to different dosage groups.Blank control group was desgined; and medicated serum groups of colchicine and Compound Biejiaruangan Tablet were taken as the positive control groups.Proliferation of HSC-T6 was detected after incubation by MTT colorimetry; cell apoptosis and the DNA content in each cell phase was detected by flow cytometer.[Results]Inhibitory rate and apoptosis rate: compared with blank control,inhibitory rate and apoptosis rate in medicated serum groups enhanced significantly,showing significant differences( P 0.01).When the dosage of medicated serum was within 5%- 20%,inhibitory rate and apoptosis rate of HSC-T6 enhanced as dosage increased.Inhibitory rate and apoptosis rate in medicated serum group of high dosage were significantly higher than those in colchicine groups,but was equal to those in Compound Biejiaruangan Tablet group.Cell cycle: there were no significant changes in cell percentage in each group at G2/M phase.Compared with blank control group,cell percentages of medicated serum groups of P.zeylanica enhanced significantly at G0/G1 phase,and reduced significantly at S phase,showing significant changes( P 0.01).When the dosage of medicated serum reduced within 20%- 5%,the cell percentage gradually declined at G0/G1 phase,and gradually increased at S phase.Under the same serum concentration,cell percentage of P.zeylanica group enhanced significantly at G0/G1 phase and decreased obviously at S phase compared with that of colchicine group.There were no significant changes in Compound Biejiaruangan Tablet group.[Conclusions] Medicated serum of P.zeylanica could restrict G1/S proliferation and induce its apoptosis,showing a dose-dependent manner.Medicated serum group of P.zeylanica had stronger effects than medicated serum group of colchicine,while had equal effects to the group of Compound Biejiaruangan Tablet.The mechanism of medicated serum of P.zeylanica in inhibiting HSC-T6 proliferation was to block cell cycle at G0/G1 phase,and to prevent it from passing G1/S.     

Abscisic acid, gibberellin and cell viability in cereal aleurone

The aleurone layer of cereals is a secretory tissue whose activity is regulated by abscisic acid (ABA) and gibberellins (GAs). Whereas GA triggers enzyme synthesis and secretion and initiates a program that culminates in cell death, ABA prevents enzyme production and cell death. Reactive oxygen species (ROS) are key players in regulating cell viability and GA sensitizes the aleurone cell to ROS. Sensitivity of GA-treated cells results in part from a reduction in steady-state amounts of mRNAs encoding enzymes that scavenge ROS. mRNAs encoding catalase, superoxide dismutase and ascorbate peroxidase are almost undetectable in aleurone layers 24 h after incubation in GA. For layers incubated in ABA, however, the amounts of these mRNAs increase. Western blotting and enzyme activity assays confirm that GA but not ABA reduced the amount and activity of ROS scavenging enzymes (Fath et al., 2001b). Substantial amounts of ROS are produced by enzymes engaged in lipid metabolism, and by the electron transport chain in the mitochondria. Aleurone layers contain abundant stores of triglycerides and ROS are produced as these lipids are rapidly converted to sugars. We hypothesize that the ROS produced in GA-treated aleurone cells bring about cell death by disrupting the plasma membrane. Aleurone cells incubated in ABA, on the other hand, are better able to maintain redox balance. ABA does not initiate rapid triglyceride metabolism, and the activities of ROS-scavenging enzymes remain high in ABA-treated cells. We conclude that GA initiates a metabolic cascade in aleurone cells that results in death from ROS. ABA maintains viability by keeping ROS under control. This revised version was published online in August 2006 with corrections to the Cover Date.     

硫化氢对白桦悬浮细胞生长、黄酮和多酚累积的影响

旨在探究硫化氢(H2S)对白桦悬浮细胞生长、黄酮和多酚等次生代谢物累积的影响。将H2S供体硫氢化钠(NaHS)添加到培养8 天的白桦悬浮培养体系中，采用pH计、电导率仪和紫外分光光度计等分析pH值、电导率、细胞活力、丙二醛、过氧化氢酶(CAT)和苯丙氨酸解氨酶(PAL)活性、黄酮和多酚含量。NaHS处理后白桦悬浮细胞培养液的pH值呈降低趋势、电导率呈增加趋势；白桦悬浮细胞中的丙二醛含量、CAT和PAL酶活性、黄酮和多酚含量呈增加趋势，而细胞活力基本呈下降趋势，且NaHS处理对上述白桦悬浮细胞生长、黄酮和多酚含量的影响存在浓度和时间效应。其中，1 mmol/L NaHS处理24 h 时黄酮和多酚含量最高，分别是对照组的128.57%和136.89%。NaHS处理促进了白桦悬浮细胞中黄酮和多酚的累积。     

植物乳杆菌CY1-2对粉红单端孢体外和番茄体内的抑菌活性及可能机制研究

以果蔬采后致病真菌粉红单端孢（Trichothecium roseum）为对象,研究了不同浓度（0、1.0×101、1.0×102、1.0×103、1.0×104、1.0×105、1.0×106、1.0×107 CFU/mL）植物乳杆菌CY1-2（Lactobacillus plantarum CY1-2）对T. roseum菌丝生长、孢子萌发和接种番茄果实腐烂情况的影响。结果表明：不同浓度的L. plantarum CY1-2均能显著抑制T. roseum菌丝生长和孢子萌发,同时也可抑制接种T. roseum番茄果实的病斑直径;浓度为1.0×105 CFU/mL的L. plantarum CY1-2处理提高了T. roseum培养液中相对电导率、可溶性糖和可溶性蛋白含量,但对丙二醛含量没有显著影响。由此表明,L. plantarum CY1-2抑制T. roseum生长的机制可能与破坏其细胞膜有关。     

2种大型海藻养殖对水体DIC含量的影响

为了解大型海藻养殖对水体溶解无机碳(DIC)含量的影响,明确其在光照、黑暗及光暗交替条件下水体DIC含量的变化过程,将取自温州洞头近海养殖的鼠尾藻、羊栖菜在实验室内进行养殖试验,设置光照、黑暗、光暗交替处理,测定了养殖水体DIC含量,并对DIC变化速率进行了分析。结果显示：在光照条件下,鼠尾藻DIC的消耗速率比羊栖菜高(P<0.05)。在黑暗条件下,鼠尾藻DIC释放速率比羊栖菜高(P<0.05)。随着光暗交替周期的延长,鼠尾藻、羊栖菜DIC的消耗速率均是先升后降,鼠尾藻DIC消耗速率比羊栖菜高。近海海藻养殖时随着光暗环境的交替进行,将发生光合作用消耗DIC、暗呼吸释放DIC的转换。总体而言,海藻养殖最终降低了水体DIC含量,能够促进大气CO₂向海水转移,而且随养殖大型海藻的收获,形成一个“可移出的碳汇”,大力发展大型海藻养殖具有重要的碳汇生态意义。     

夏玉米籽粒胚乳细胞增殖及产量对不同光照的响应

为了探讨不同光照条件对夏玉米籽粒胚乳细胞增殖及产量的影响,2012—2014年于山东农业大学试验农场进行了大田试验,选玉米品种郑单958和登海605为试验材料,设置大田自然光照(CK)、开花至收获期遮阴(S1)、拔节至开花期遮阴(S2)、出苗至收获期遮阴(S3)和开花至收获期增光(L)5个处理,研究其对夏玉米籽粒胚乳细胞增殖、籽粒灌浆、淀粉含量、干物质积累及产量的影响。结果表明,遮阴后籽粒胚乳细胞增殖速率、细胞数目和淀粉积累量降低,细胞充实度下降,粒重显著降低,且S3对其影响最大,S1次之,S2影响相对较小。郑单958和登海605两个品种S1、S2和S3成熟期的胚乳细胞数目较CK分别降低33%、6%、29%和41%、5%、29%,DH605 L处理显著提高了胚乳细胞增殖速率、胚乳细胞数目及淀粉含量,细胞充实度和粒重显著上升。遮阴导致籽粒灌浆速率减缓,灌浆高峰期推迟,2个品种S1、S2和S3的最大灌浆速率较CK分别降低34%、13%、58%和38%、13%、64%,登海605L处理提高了籽粒灌浆速率,籽粒灌浆高峰期提前出现。2个品种S1、S2和S3较CK分别减产58%、26%、81%和67%、27%、81%,登海605 L处理增产9%。即遮阴降低了籽粒胚乳细胞数目、淀粉含量和灌浆速率,进而影响产量,花粒期充足的光照则有助于籽粒胚乳细胞增殖和产量提高。     

Responses to Aluminium Toxicity Effects of Two Populations of Piptatherum miliaceum (L.) Cosson

The effects of different aluminium (Al) concentrations (0 (control), 5 and 25 ppm) and pH levels (4.5 and 10.0) on two Piptatherum miliaceum populations, one from a bauxite area (population 1) and the other from pasture soil (population 2) were studied. Root length of both populations was severely inhibited at 25 ppm Al concentration and pH 4.5. Positive correlations between the Al content of roots of the two populations and increasing Al concentrations and pH levels in the nutrient solution were observed. Increase of Al concentrations in nutrient solution resulted in the decrease of calcium (Ca) content of the roots of the two populations in both pH values, while potassium (K) concentrations in the roots slightly differed from those of the control. Leaf chlorophyll content increased with increasing Al concentrations and pH values of nutrient solution in population 1 but decreased in population 2.
Based on relative values at different treatments of root length (R.L.), Ca and K content of roots and leaf chlorophyll content, it is concluded that population 1 is more tolerant to Al than population 2.
The chlorophyll content, under Al influence, may serve to distinguish Al-tolerant and non-tolerant ecotypes.     

Inhibitory Effects of Polyphenols from Pinus koraiensis Seed Scale on the Proliferation of Human Cancer Cells in Vitro

[Objective] To research on the inhibitory effects of polyphenols from Pinus koraiensis seed scale on the proliferation of human cancer cells in vitro. [Methods] Polyphenols from Pinus koraiensis seed scale were prepared into extracting solutions at different concentrations. Suspension cultures of five tumor cells were processed,which were lung carcinoma cell A549,human bone marrow neuroblastoma cell SH-SY5 Y,human skin cancer cell A375,human hepatocarcinoma cell HepG-2 and human ovarian cancer cell SKOV3. Inhibitory rate of cell proliferation in vitro was detected by MTT method. [Results] P. koraiensis polyphenols had relatively good inhibitory effects on A549,showing certain correlation with time or concentration. The inhibitory rare was the optimal when the concentration of P. koraiensis polyphenols was0. 4 mg / mL. Under this concentration,the inhibitory rate of extracting solution of P. koraiensis polyphenols reached 55% on lung carcinoma cell A549. P. koraiensis polyphenols showed no significant inhibitory effects on SH-SY5 Y,A375,HepG 2 or SKOV3. [Conclusions] P. koraiensis polyphenols had inhibitory effects on lung carcinoma cells.