Epidermal collagenase activity and its induction by 20-hydroxyecdysone in the fiddler crab Uca pugilator

The epidermal collagenase activity and its induction by 20-hydroxyecdysone in Uca pugilator were investigated.Zymographic electrophoresis showed four bands of collagenae activity,16,19,22 and 29 kDa in molecular weight,with the former two accounting for 60% and 36%,respectively,of the total coUagenase activity. The collagenase activity varies during the molting cycle. Among the molt stages tested,Premoh Stage Do exhibited the highest epidermal coUagenase activity for both the 16 and 19 kDa isoenzymes and,as the molt stage proceeded,the enzymatic activity of these two isoenzymes decreased,with the lowest activity for both found in Premoh Stage D3-4. Injection of 20-hydroxyecdysone significantly induced the activity of the 16 kDa collagenase in the epidermis of Uca pugilator,suggesting that the activity of this isoenzyme is under molting hormone control.Although 20-hydroxyecdysone injection did not result in a statistically significant increase in the activity of the 19 kDa isocnzyme,a tendency of the induction was nonetheless demonstrated. This is the first report on epidermal collagenase activity and its induction by the molting hormone in a crustacean.     

Sorcin蛋白高表达对胃癌细胞生长的影响

目的：研究Sorcin蛋白在胃癌及癌周组织中的表达及其在胃癌发生发展中的可能作用机制。方法：应用免疫组化检测Sorcin在85例胃癌及癌周组织中的表达情况;转染Sorcin基因至正常胃粘膜上皮细胞GES-1,Western blot验证转染组及对照组Sorcin表达情况;MTT法检测细胞存活率。结果：Sorcin在胃癌组织中的表达明显高于癌旁组织（p＆lt;0.01）;与对照组相比,Sorcin高表达明显增强细胞抗凋亡能力。结论：Sorcin蛋白质在胃癌中高表达,其可能通过调控细胞凋亡途径参与胃癌恶性生物学行为,有望成为潜在的诊断胃癌、判断预后的分子标志物。     

ocs/mas|一个受损伤和植物激素诱导的嵌合启动子的构建与功能分析

选择适宜的转录调控序列以提高启动子的转录效率,增强外源基因在转基因植株中的表达,对改良作物的抗病虫性具有重要意义。将甘露碱合成酶基因(mas)启动子和章鱼碱合成酶基因(ocs)增强子杂合而成的嵌合启动子ocs/mas与GUS报告基因连接,构建了植物表达载体pOMS-GUS。对照载体pMAS-GUS仅携带mas启动子驱动的GUS基因。利用根癌农杆菌介导法,将以上植物表达载体分别转化烟草。应用半定量RT-PCR和GUS荧光定量分析法分别检测不同胁迫条件下启动子驱动的GUS基因表达量的变化。结果显示,未诱导处理的转基因植株GUS基因仅有微弱表达。伤害处理1h后,mas启动子驱动的GUS活性是未诱导处理的1.8倍,而嵌合启动子ocs/mas的诱导表达活性是未处理的5.7倍。植物激素水杨酸(SA)和茉莉酸甲酯(MJ)处理也诱导了较高水平的ocs/mas嵌合启动子活性;而且SA和MJ联合作用时呈现叠加效应,转基因烟草的GUS活性明显高于伤害处理后的GUS表达水平。以上结果表明,ocs/mas嵌合启动子是一种强诱导型启动子,可以接受多种刺激因子的诱导,从而为更有效地改良作物抗病虫的能力提供新的候选高效启动子元件。     

p300通过homeobox结构域增强Nanog的转录激活活性

胚胎干细胞(ES细胞)能够不断地进行自我更新来维持其多能性,很多转录因子共同调控着ES细胞的自我更新和多能性,Nanog就是其中之一,然而Nanog维持ES细胞多能性的机制并不清楚。p300是真核生物中普遍存在的转录辅助因子,与许多转录因子共同作用调控下游基因的表达。为探索p300是不是能够影响Nanog的转录活性,我们在细胞中共转Nanog(或突变体)及报告基因和p300,结果表明p300能够通过homeobox结构域增强Nanog的转录激活活性。     

炭团菌提取物对樟子松枯梢病菌的抑菌活性及稳定性研究

采用水提和酯提2种方式提取炭团菌液体培养菌液中高效抑制樟子松枯梢病菌活性物质。采用生长速率法测定提取物对病原菌菌丝生长的抑制率,采用悬滴法测定其对病原菌孢子萌发的抑制率。炭团菌液体培养菌液乙酸乙酯提取物稳定性试验选用紫外线、温度、pH、氧化剂与还原剂以及贮存时间5个因子。结果表明:炭团菌液体培养菌液的5种提取物对樟子松枯梢病菌均有一定的抑菌活性,乙酸乙酯提取物对病原菌菌丝生长的抑制率最高,达到73.20%;乙酸乙酯提取物、超声波提取物、正丁醇提取物对病原菌孢子萌发抑制率均超过90%。乙酸乙酯提取物对自然环境下的紫外线、温度具有很高的稳定性,并且有一定的抗氧化还原能力和良好的耐热性,长期贮存不影响其抑菌活性,具有很高的开发价值和很好的应用前景。     

油菜花粉总黄酮的微波辅助提取及其抗氧化活性研究

采用微波辅助提取法提取油菜花粉总黄酮,通过单因素和正交试验,得到最佳提取条件：80％的乙醇、1：20的料液比、提取时间140s、微波功率242W、提取3次,总黄酮质量分数为（2．64±0．05）％。同时还研究提取物对羟自由基、DPPH自由基的清除作用,以及对小鼠肝组织脂质过氧化的抑制作用,自由基半清除质量浓度和脂质过氧化丰抑制盾奄浓度分剐为0．115、0．325和0．065mg／mL。     

干旱胁迫条件下小麦旗叶酶活性和丙二醛含量的染色体定位

以中国春-Synthetic 6x小麦染色体代换系及其亲本为材料,在不同生育时期对其旗叶SOD、POD活性和丙二醛(MDA)含量进行测定.结果表明,在干旱胁迫下,2B和7D代换系叶片的SOD活性及其相对活性在孕穗期、开花期和灌浆期始终显著或极显著高于中国春;1A、2A和2D代换系叶片POD活性及其相对活性始终显著或极显著高于中国春;而7A、1D和7D代换系叶片的MDA含量及其相对MDA含量始终显著或极显著低于中国春.由此表明,Synthetic 6x的2B和7D染色体上可能存在干旱胁迫下诱导SOD活性增强的有利基因;1A、2A和2D染色体上可能存在诱导POD活性增强的有利基因;7A、1D和7D染色体上可能存在抑制MDA含量增高的基因.     

紫芝酸性三萜类化合物体外抑癌和抑菌作用的研究

采用噻唑蓝比色(MTT)法研究紫芝酸性三萜对几种癌细胞体外增殖的影响,并用管碟法检测了紫芝胞内酸性三萜对几种细菌和霉菌的体外抑菌作用。结果表明,紫芝胞内酸性三萜和胞外酸性三萜在250μg/mL时,对人肝癌细胞BEL7402和人乳腺癌细胞MCF-7均有显著抑制作用(P<0.05),但对人胃癌细胞SGC-7901没有显著抑制作用(P>0.05)。BEL7402细胞的生长曲线试验表明,胞内酸性三萜组的细胞受到显著抑制,未出现指数增长期,且BEL7402细胞培养3d后,对照组细胞数目多、均匀,而胞内酸性三萜组的细胞数目明显减少,且细胞变小。抑菌试验结果表明,胞内酸性三萜在40mg/mL时,对大肠杆菌Escherichia coli和金黄色葡萄球菌Staphylococcus aureus的生长均具有显著抑制作用(P<0.01),对枯草芽孢杆菌Bacillus subtitis和青霉的Penicillium chrysogenum抑制作用较弱;而在此浓度下对黑曲霉Aspergillus niger没有抑制作用。该样品对大肠杆菌、金黄色葡萄球菌、枯草芽孢杆菌、黑曲霉和青霉的MIC分别为20mg/mL、20mg/mL、40mg/mL、80mg/mL和40mg/mL。此外,该酸性三萜的抑菌成分在60℃下(处理2h)较稳定,但在80℃以上,热稳定性较差,活性降低。     

盐渍条件下AM真菌对大豆生长和离子含量的影响

盐渍条件下研究了丛枝菌根(AM)真菌对大豆Glycine max植株生长和叶片离子含量的影响。结果表明,接种摩西球囊霉Glomus mosseae处理的叶片K+含量和K/Na比显著高于对照,而Na+含量无显著差异。G.mosseae显著增加了大豆植株生长量,这一效应随盐处理浓度的提高而增大。表明盐渍条件下AM真菌提高大豆抗盐性与其增加K+吸收和运输有关。     

Evaluating the antitumor activity of combined photochemotherapy mediated by a meso-substituted tetracationic porphyrin and adriamycin

The combined anticancer modality comprising porphyrins as photodynamic sensitizers and anticancer drugs has been an interesting subject for many researchers. In this study, the photochemotherapeutic effect mediated by simultaneous photoactivation of tetracationic meso-tetrakis（N-methyl-4-pyridyl） porphine tetratosylate （TMPyP） and adriamycin （ADM） were explored using human hepatocellular carcinoma cell line （HePG2）. The efficiency of TMPyP acting in concert with ADM in the dark and in the presence of photoirradiation was evaluated, by studying cell viability, caspase-3 activity and ultrastructural changes in the cells after incubation with each of the two agents, separately, or simultaneously as a co-mixture. Under dark conditions, the simultaneous incubation of cells with TMPyP and ADM significantly enhanced cell death by 1.8 folds and 1.3 folds, compared with TMPyP or ADM treatment, respectively. After photoirradiation, the antiproliferative effect of the co-treatment with TMPyP and ADM increased further by 2 folds. Transmission electron microscopy and the measurements of caspase-3 levels in treated cells revealed that the co-treatment of cells with ADM and TMPyP followed by light irradiation directed the cell death towards necrosis and abrogated the apoptotic cell death pathway, which was exhibited in cells treated with ADM in absence and in presence of photoirradiation.