半夏多倍体复合体基因组DNA甲基化状态的MSAP分析

目的分析半夏多倍体复合体DNA甲基化状态的变化(甲基化水平和甲基化差异模式)。方法应用甲基化敏感扩增多态性(methylation sensitive amplified polymorphism,MSAP)技术,采用34对引物进行选择性扩增。结果扩增共得到7708条带,其中5636条带为甲基化多态性带在半夏7倍、8倍、9倍、10倍体值株间DNA甲基化水平变化不大。总扩增位点甲基化水平在54%～58%,全甲基化率为24.1%～24.3%,高倍性植株甲基化程序相应比较高。除甲基化水平稍有变化外,半夏不同倍性株间的DNA甲基化模式也形式各异,检测到的带型分为两大类(和型)。其中,不同倍性间DNA甲基化状态保持不变的位点占52.5%,归为型;少部分检测位点(占47.5%,归为型)的DNA甲基化模式在不同倍性间存在显著差异。结论半夏多倍体复合体各倍性植株间存在大量的胞嘧啶甲基化变异且整体甲基化水平较高。     

不同干燥工艺对鱼腥草挥发油的影响研究

目的研究不同干燥工艺对大别山鱼腥草挥发油的影响。方法分别用低温烘干(50℃)、晒干、阴干的干燥方式处理新鲜的鱼腥草,挥发油提取器提取挥发油,采用GC-MS检测分析。结果阴干、晒干、烘干鱼腥草中的挥发油分别鉴定出43、37和14种。结论晒干和低温烘干方式均会影响鱼腥草挥发油的组成,但对总体积影响不显著。     

Effects of Ixeris Chinensis (Thunb.) Nakai Boiling Water Extract on Hepatitis B Viral Activity and Hepatocellular Carcinoma

Background

Hepatitis B virus (HBV) infection and hepatocellular carcinoma are major diseases that affect the Taiwanese population. Therefore, the development of an alternative herbal medicine that can effectively treat these diseases is a research target. In this study, we tested Ixeris Chinensis (Thunb.) Nakai boiling water extract (ICTN BWE) in vitro and analysed its effects on the HBV and liver cancer.

Materials and Methods

We used a human liver cancer cell line (Hep3B, a cell line that continuously secretes HBV particles into a medium) as an experimental model for the screening of various ICTN BWE concentrations and their effects on the HBV in vitro.

Results

Our results showed that 75 µg/mL ICTN BWE downregulated the relative expression of the hepatitis B virus surface antigens (HBsAg) to 77.1%. Using the human liver cancer cell lines HuH-7 and HepG2, and 3-(4,5-dimethylthiazol-zyl)-2,5-diphenyl tetrazolium bromide (MTT) and tumour clonogenic assays, we then showed that ICTN BWE inhibits hepatocellular carcinoma growth.

Conclusion

Fluorescent microscopy of DAPI(4′,6-Diamidino-2-phenylindole)-stained nuclei and DNA fragmentation assays confirmed the inhibitory effects of ICTN BWE on liver tumour cell growth through induction of apoptosis.     

全基因组水平金银花TCP基因家族的鉴定及表达模式分析

目的 对金银花Lonicera japonica TCP（LjTCP）基因家族进行鉴定分析,以期为进一步研究LjTCPs的功能机制奠定基础。方法 以金银花基因组数据为基础,通过生物信息学系统分析TCP基因家族在染色体上的分布、系统进化、基因结构、启动子元件及其表达模式。结果 共鉴定出17个LjTCPs,其中16个LjTCPs分布在9条染色体上,LjTCP17未定位到染色体上,基于系统进化树和多重序列比对,LjTCP基因家族可以分为PCF、CIN和CYC/TB1 3个亚家族,分别包含9、6、2个LjTCPs。基因组内共线性分析表明,全基因组复制和片段性复制在LjTCP基因家族进化中发挥了重要作用。LjTCP基因家族启动子包含许多与植物生长发育、激素响应,以及非生物和生物胁迫相关的调控元件。表达模式分析表明,LjTCP基因在金银花花发育初期表达量较高,随着发育进程表达量呈现下降趋势,并且LjTCP05、LjTCP13、LjTCP14、LjTCP16和LjTCP17基因在花青素含量不同的品种中表达量存在差异。对6个具有低温响应元件的LjTCPs在冷胁迫下的基因表达研究表明,大部分基因在冷处理后基因表达呈现上升趋势。结论 金银花TCP基因家族包含17个成员,各成员的分子特征和表达模式存在差异。     

忍冬VIGS基因沉默体系构建

目的 以忍冬Lonicera japonica八氢番茄红素脱氢酶（phytoene desaturase,PDS）基因为标记基因,利用ZMBJ-CMV载体建立基于病毒诱导的忍冬基因沉默体系。方法 以忍冬为材料,通过选取同源性较高的片段设计PDS基因的特异性引物,克隆出片段长度为135 bp的序列作为目的基因片段,经双酶切后与黄瓜花叶病毒载体连接构建VIGS重组载体,转化根癌农杆菌后,侵染忍冬幼苗。此外,还对农杆菌吸光度（A）值和侵染方法对PDS基因沉默效率的影响进行了比较。结果 被侵染植株叶片PDS基因表达水平显著降低,叶绿素和类胡萝卜素含量显著下降,叶片出现白化表型。结论 建立了以ZMBJ-CMV载体诱导的忍冬基因沉默体系,农杆菌A值为0.8、注射后再进行真空浸润,能够提高忍冬目标基因的沉默效率,为开展忍冬关键基因的功能研究提供了技术支撑。     

HPLC法测定何首乌及复方虫草胶囊中大黄素、大黄素甲醚的含量

采用 HPL C法测定了何首乌及含首乌制剂复方虫草胶囊中游离及结合型大黄素、大黄素甲醚的含量。色谱柱 :Waters型 C1 8柱 (10μm,2 5 0 m m× 4.6 mm ) ;检测波长 :2 5 4nm;流动相 :甲醇 -水 -磷酸 (85 0∶ 15 0∶ 1) ;流速 :1m L/ min。方法简便易行 ,结果准确 ,重现性好 ,可有效地控制生药及其制剂的质量     

Effects of Polysaccharides from Gynostemma Pentaphyllum (Thunb.), Makino on Physical Fatigue

Background

Gynostemma pentaphyllum (Thunb.) Makino has been reported to have a wide range of health benefits in Chinese herbal medicines. Polysaccharides from Gynostemma pentaphyllum (PGP), has been identified as one of the active ingredients responsible for its biological activities. Although many pharmacological activities of PGP have received a great deal of attention, there is limited evidence for the anti-fatigue effects of PGP. The purpose of this study was to investigate the effects of polysaccharides from PGP on physical fatigue.

Materials and method

The rats were divided into four groups, with 10 animals per group: control (C), group, low-treated (LT), group, medium-treated (MT), group, and high-treated (HT), group. The C group received distilled water, while LT, MT and HT groups were given various doses of PGP (100, 200, 400 mg/kg· d). After 30 days, forced swimming test was carried out in an acrylic plastic pool, then the exhaustive swimming time of rats and some biochemical parameters related to fatigue were measured. The data obtained showed that PGP could extend the exhaustive swimming time of the rats, as well as decrease the blood lactic acid (BLA), and blood urea nitrogen (BUN), concentrations, and increase the hemoglobin, liver glycogen and muscle glycogen concentrations.

Result

The data obtained showed that different doses of PGP could extend the exhaustive swimming time of the rats, as well as decrease the BLA and BUN concentrations, and increase the hemoglobin, liver glycogen and muscle glycogen concentrations, which suggests that PGP had significant anti-fatigue effects on rats.

Conclusion

PGP may be of use as a potential anti-fatigue agent, but there is a need for further research on long-term use in order to show its positive effects on physical fatigue.     

壮药赪桐乙酸乙酯部位及流份对LPS诱导的RAW264.7细胞的抗炎作用

目的 筛选赪桐乙酸乙酯部位及不同洗脱梯度二氯甲烷-甲醇部位对炎症反应的抑制作用最强的流份。 方法 使用MTT法确定壮药赪桐乙酸乙酯部位及不同洗脱梯度二氯甲烷-甲醇部位对RAW264.7细胞安全给药浓度范围,通过ELISA法测定赪桐乙酸乙酯部位及不同洗脱梯度二氯甲烷-甲醇部位对LPS诱导RAW264.7细胞分泌NO、TNF-ａ、IL-12、IL-6、IL-1β含量,筛选对炎症反应的抑制作用最强的流份。 结果 赪桐乙酸乙酯部位及流份在0.06~2.0 mg/ml浓度范围内,赪桐乙酸乙酯部位及流份对细胞活力的抑制作用逐渐增强,浓度在0.5 mg/ml以上具有明显的细胞毒性,在0.5 mg/ml以下对细胞活力具有增强作用。二氯甲烷-甲醇洗脱部位高剂量能抑制炎症因子IL-12、IL-6、TNF-ａ、IL-1β释放,对NO的分泌没有抑制作用。 结论 赪桐乙酸乙酯部位及不同洗脱梯度的二氯甲烷-甲醇部位抗炎机制是通过抑制NO、TNF-ａ、IL-12、IL-6、IL-1β炎症因子的分泌,二氯甲烷-甲醇（50:1）洗脱部位和二氯甲烷-甲醇（30:1）洗脱部位的抗炎能力较强。     

A Study on Anti-Tumour Effect of Solanum Lyratum Thunb. Extract in S180 Tumour-Bearing Mice

The objective of the study was to investigate the anti-tumour effect of ethanol extract of Solanum lyratum Thunb. in S₁₈₀ tumour-bearing mice, and to preliminarily explore its mechanism of action. Methods: Mice were made into S₁₈₀ solid tumour model, grouped and administered. Tumour inhibition rate was measured by harvesting the tumours. Serum IL-2, TNF-a contents were measured by taking blood samples, and thymus index and spleen index were measured by harvesting the thymus and spleen. The results showed that the Solanum lyratum Thunb. extract had certain tumour inhibitory effect, which can elevate the serum IL-2, TNF-a contents, and increase the thymus and spleen indices to a certain extent. The study concluded that Solanum lyratum Thunb. extract has certain in vivo anti-tumour effect which may be exerted through enhancing the body immunity.