灰绿碱蓬多糖脱蛋白工艺研究

蛋白本身具有热敏感性,糖蛋白复合物在脱除蛋白质以后,多糖自身的生物活性会明显增加。本文测定了灰绿碱蓬根、茎、叶的基本营养成分,采用水提醇沉法提取了灰绿碱蓬多糖,并优化了多糖脱蛋白工艺比较了Savage法、(NH4)SO4沉淀法、TCA法和酶-Savage法这4种方法的脱蛋白效果。结果显示,灰绿碱蓬多糖的最佳脱蛋白方式为酶-Savage法,酶添加量为7%,Savage溶液处理一次,此条件下脱蛋白效果和多糖保留率均最好,蛋白质脱除率为80.71%,多糖保留率为67.27%。     

Induction of apoptosis with Salvia miltiorrhiza monomer IH764-3 via downregulating focal adhesion kinase in H2O2-stimulated hepatic stellate cells

AIM: To investigate the effect of IH764-3 on the apoptosis of H₂O₂-stimulated hepatic stellate cells(HSCs) and the alteration of focal adhesion kinase (FAK). METHODS: The proliferation of HSCs was examined by direct cell count and the apoptosis was determined by Annexin-V/PI labeling, while the morphological change was observed by light microscopy and transmission electron microscopy. In addition, FAK mRNA was detected by RT-PCR. RESULTS: H₂O₂ promoted the proliferation of HSCs. IH764-3 induced the apoptosis of HSCs in a dose-dependent manner. The HSC apoptotic rates of different groups were 6. 35%,9. 28%,15. 10%,19. 69%,respectively, after treated with different concentrations of IH764-3 for 48 h while H₂O₂ group showed 2. 30%. In 30 mg/L group, the apoptosis rates were 6. 73%、10. 34%、15. 10% for the indicated time periods(12 h, 24 h, 48 h). In the presence of IH764-3, FAK mRNA decreased. The FAK mRNA reduction began at 2 h after adding IH764-3. CONCLUSION: IH764-3 induced the apoptosis of HSCs. Down-regulating the expression of FAK mRNA may be one of its mechanisms.     

南欧丹参花提取物杀菌活性初步研究

采用系统溶剂对南欧丹参Salvia sclarea L.花进行提取,以孢子萌发试验法、抑制菌丝生长速率法和盆栽试验系统测定了粗提物对植物病原真菌的生物活性。结果表明,石油醚为提取南欧丹参花活性成分的最佳溶剂。石油醚提取物在离体条件下能显著抑制多种供试真菌菌丝的生长,其对番茄灰霉病菌Botrytis cinerea的EC50值为77.20 μg/mL;对盆栽小麦白粉病Erysiphe graminis的防护效果为94.94%,治疗效果为84.21%。     

Clinical,pathologic, and toxicologic characterization of Salvia reflexa (lance-leaf sage) poisoning in cattle fed contaminated hay

Salvia reflexa (lance-leaf sage)-contaminated alfalfa hay was fed to ~500 mixed-breed beef cattle. Within hours of exposure, nearly half of the cattle developed lethargy, anorexia, depression, and recumbency, followed by bellowing, colic, and death. Even though the uneaten contaminated hay was removed the first day, nearly 100 animals died within the first 48 h. Three of these cattle were examined postmortem, and tissues and hay samples were collected for microscopic and chemical analysis. Several days later, a smaller number of the clinically poisoned cattle developed neurologic disease with aberrant behavior, aggression, icterus, blindness, exhaustion, and death. A total of 165 cattle were fatally poisoned. Poisoned cattle had swollen, dark, mottled livers that had a prominent nutmeg-like lobular pattern on cut section. Histologically, there was severe centrilobular-to-panlobular hepatic necrosis with marked hepatocellular swelling, degeneration, and necrosis. The surviving cattle developed liver disease characterized by altered serum biochemical analyses and microscopic hepatocellular degeneration and necrosis. In subsequent biopsies and analysis, these lesions resolved within 6–7 mo. After confirming toxicity of the hay in cattle, goats, and mice, followed by a mouse bioassay–guided chemical fractionation process, Salvia reflexa was identified as the contaminant in the hay responsible for the hepatotoxicity. S. reflexa has not been reported previously to cause fatal hepatotoxicity in livestock in North America, to our knowledge.     

铜、锌离子影响丹参金属硫蛋白MT2基因表达

植物金属硫蛋白在金属离子的储存、运输和代谢及降低重金属离子的毒性等方面起着重要作用本文研究了Cu2 、Zn2 对丹参毛状根中金属硫蛋白基因表达的影响.以6,7-V为液体培养基,分别以缺乏正常及原培养基100、500、1 000倍的cu2 、Zn2 处理,采用半定量PCR法检测不同时间点时金属硫蛋基因的表达情况.结果发现Cu2 、Zn2 诱导金属硫蛋基因的表达量分别在100倍处理量时的9 h和6 d达到峰值Cu2 、Zn2 能显著诱导丹参金属硫蛋白基因的表达,该基因可能在丹参对铜、锌元素的吸收方面发挥着重要的作用.     

梭梭幼苗根系分泌物提取方法的研究

为了探讨梭梭根系分泌物中的化学成分,本文采用水浸提取和循环灌溉两种方法对梭梭幼苗根系分泌物进行收集和提取,结合气相色谱-质谱联用分析技术,分析梭梭幼苗根系分泌物中组分和含量的差异。实验结果表明:循环灌溉收集的梭梭幼苗根系分泌物中检测到35种化合物,包括酚醌类、醛酮类、醇类、酸类、脂类和胺类化合物;硅烷化处理后,主要增加部分强极性有机酸,如苯甲酸。水浸提取梭梭幼苗根系分泌物的中性相组分共检测出27种化合物,但未检测到酚醌类化合物;碱性相组分共检测出23种化合物,醛酮类化合物种类减少,烷烃类化合物种类增加;酸性相组分中仅检测到10种化合物,以醇类化合物为主,占检出化合物总量的72.62%。收集梭梭幼苗根系分泌物循环灌溉是一种有效的方法。     

丹参海藻糖-6-磷酸合成酶基因（SmTPS）的克隆及其表达分析

对丹参EST序列进行Blast分析,得到一条海藻糖-6-磷酸合成酶（TPS）基因,将其命名为SmTPS,GenBank注册号为JF937196。该基因cDNA全长2836 bp,包含一个长为2574 bp的ORF,编码857个氨基酸。生物信息学分析表明,分子量为97.04 kD,等电点为5.76,含α-螺旋、β转角、延伸连和无规则卷曲。该蛋白同时具有海藻糖-6-磷酸合成酶（TPS）和海藻糖-6-磷酸磷酸酯酶（TPP）的功能结构域。系统进化树分析表明丹参TPS功能域和拟南芥AtTPS6属于一类,而TPP功能域与低等生物酵母ScTPS2属于一类。实时定量PCR结果分析表明,SmTPS基因在丹参不同组织器官中差异表达,其茎中表达量最高。在干旱和低温处理下,其表达量与对照相比均可上调约2倍,表明SmTPS基因在丹参抵抗干旱和低温胁迫中发挥一定的作用。     

丹参DHN2基因的克隆与序列分析

本文以组织培养2～13周的丹参幼苗为材料，构建丹参cDNA文库并进行大规模EST序列分析，所得序列经NCBI的BLAST工具分析，克隆号为rsmsxl_014903．y1．scf序列与晚期胚胎丰富(late embryogenesis abundant)基因家族Ⅱ中的成员有较高的同源性。该序列全长864bp，包含1个长726bp开放阅读框(ORF)，编码241个氨基酸，并含有ⅡLEA蛋白的特征序列，与NCBI注册的脱水素家族基因具有较高的同源性，表明本基因可能是一种新的脱水素基因，命名为DHN2，并在GenBank上进行了注册，序列号为：AY737725。     

氮、磷、钾肥对丹参根系生长及养分含量的影响

采用田间小区试验方法,研究了氮、磷、钾肥对丹参根系生长及养分含量的影响。结果表明,氮、磷、钾肥能够促进丹参根系对氮、磷、钾的吸收,提高其含量,并且随着施用量的增加而提高,在N,P2O5,K2O的施用量分别为225,180,225 kg/hm2时,根的氮、磷、钾含量达到最高值;当施入N,P2O5和K2O分别为225,120,150 kg/hm2时,丹参根干质量、根长及根条数达到最高值,根的直径最适宜,此后随着磷钾肥施入量增加,根干质量及根长、根条数反而下降,根直径增加。所以氮、磷、钾合理配合施用能够提高丹参根的养分含量、根干质量、根长、根数及根直径。     

小根蒜提取物对香椿保鲜效果的影响

通过测定Vc、叶绿素含量变化并结合外观变化,研究了小根蒜提取物对香椿保鲜效果的影响。结果表明：小根蒜提取物对香椿具有显著的保鲜效果,可以延长保鲜期7天以上。小根蒜水提物可以显著降低香椿的腐烂率,抑制Vc含量的下降,贮藏28天香椿的完好率仍维持在89.6%,Vc保持率为49.4%。小根蒜醇提物与6-BA复配可有效降低香椿的脱叶率和失重率,延缓叶绿素的降解,贮藏28天脱叶率为20.3%,失重率和叶绿素损失率分别为3.8%和5.3%。以小根蒜提取物作为天然蔬菜保鲜剂具有巨大的开发潜力。