Daphne-type diterpenes from genus Daphne and their anti-tumor activity

Daphne-type diterpenenoids is the major biologically active constituents in the genus Daphne. We find that there are about 101 Daphne-type diterpenes in this genus, most of these compounds show different degrees of inhibitory effect on various cancer cell. Some of them have been studied in depth and the potent molecular mechanisms might be associated with modulation of different cell-signaling pathways. In addition, some compounds of this type also can inhibit the synthesis of protein and DNA. Absolutely, the anti-tumor activity of Daphne-type diterpenes is worthy of attention. Unfortunately, most of the current research on the activity of these compounds is focused on simple drug efficacy, and its in-depth mechanism research is far from enough. On the other point of view, there still exists wide growing space on the depth of these compounds.     

基于网络药理学的黄瑞香抗胃癌作用机制探讨及实验验证

目的 采用网络药理学方法预测黄瑞香发挥抗胃癌作用的潜在机制,并通过体外细胞实验进行初步验证。方法 利用中药系统药理学数据库和分析平台（TCMSP）筛选黄瑞香的有效成分,通过Swiss Target Prediction数据库和PharmMapper数据库筛选获得黄瑞香活性成分的相关靶点。通过GeneCards及DisGeNET数据库检索获得胃癌疾病相关靶点。利用Cytoscape软件构建黄瑞香活性成分-靶点网络;通过STRING、Metascape数据库对成分和胃癌疾病交集靶点进行蛋白质相互作用（PPI）网络和基因本体（GO）注释及京都基因与基因组百科全书（KEGG）通路富集分析。采用MTT实验考察黄瑞香异戊烯基黄酮成分构树黄酮醇F（broussoflavonol F）、daphnegiravone D、daphgiflavone C体外对胃癌MGC803细胞生长的影响,采用实时荧光定量PCR方法检测这3种异戊烯基黄酮类化合物作用48h后胃癌MGC803细胞表皮生长因子受体（EGFR）、磷脂酰肌醇3-激酶（PI3K）和蛋白激酶B（Akt）基因的表达。结果 筛选得到黄瑞香70个活性成分,黄瑞香作用于胃癌的42个相关靶点,其中表皮生长因子受体（EGFR）、鼠肉瘤病毒癌基因同源物B1（BRAF）、Ⅰ类磷脂酰肌醇-3激酶催化亚基α（PIK3CA）、酪氨酸激酶（MET）以及丝裂原活化蛋白激酶1（MAPK1）等为核心靶点。KEGG富集分析发现黄瑞香作用于胃癌主要富集在癌症、PI3K/Akt以及丝裂原活化蛋白激酶（MAPK）等通路中。活性测试结果发现,异戊烯基黄酮成分构树黄酮醇F、daphnegiravone D、daphgiflavone C对胃癌MGC803细胞生长具有较好的抑制作用。同时,与对照组比较,构树黄酮醇F、daphnegiravone D、daphgiflavone C可显著抑制胃癌MGC803细胞EGFR、PI3K和Akt基因的表达（P＜0.05、0.01、0.001）。结论 黄瑞香可能以异戊烯基黄酮为主要的抗胃癌活性成分,以EGFR、BRAF、PIKC3A、MET等蛋白为核心作用靶点,主要经I3K/Akt以及MAPK等相关信号通路发挥治疗胃癌的作用。     

芫花中抗肿瘤活性成分瑞香烷型二萜的含量测定研究

目的:建立HPLC法同时测定不同产地芫花药材中7个具有抗肿瘤活性的瑞香烷型二萜[yuanhuadine(1),yuanhuafine(2),genkwadaphnine(3),yuanhuacine(2),yuanhuahine(5),yuanhualine(6),isoyuanhuadine(7)]的含量。方法:芫花以75%乙醇水溶液提取;色谱柱为Zorbax Eelipse SB-C18(250 mm×4. 6 mm,5μm),流动相:乙睛-0. 1%冰醋酸水,检测波长:230 nm,柱温:40℃,流速:0. 8 m L/min,进样量:10μL,洗脱梯度:采用二元梯度洗脱系统,溶剂A(乙睛),B(水、0. 1%冰醋酸)。结果:上述7个瑞香烷型二萜的线性关系良好(r≥0. 9941),平均加样回收率为98. 1%～104. 5%。不同生长环境的芫花中的7个成分各自含量有所差别。结论:该方法可用于中药芫花中瑞香烷型二萜的含量测定研究。     

回收率检验-正交函数分光光度法及其程序设计

本文提出的回收率检验-正交函数分光光度法,减小了在中药分析上应用正交函数法的难度,借助于电子计算机,使得最适实验条件的选择变得简单易行。通过芫花根注射液有关数据的计算,得到了7组回收率在99～101%范围内的适宜条件,8组回收率在98～102%范围内的适宜条件。     

正交函数分光光度法测定芫花根注射液中二萜原酸酯类的含量

芫花根注射液是一种粗制剂,成分复杂,含量测定困难,其主要引产成分为芜花酯甲,芫花酯乙,芫花酯丙和芜花酯丁等瑞香烷型二萜原酸酯类,其中以芫花酯甲含量最高。本文以芫花酯甲为标准品,用正交函数分光光度法,测定芫花根注射液中总瑞香烷型二萜原酸酯的含量。其平均回收率为98.9%,变异系数1.6%(n=9)。     

唐古特瑞香抗炎镇痛活性部位筛选研究

目的筛选出唐古特瑞香抗炎镇痛的活性部位。方法采用醋酸扭体法、小鼠腹腔毛细血管通透性试验以及二甲苯致小鼠耳肿胀试验,分别以小鼠扭体反应次数、毛细管通透性、小鼠耳肿胀度以及胀抑制率为考察指标,对唐古特瑞香的乙醚部位、50%乙醇部位、30%乙醇部位、水部位进行抗炎镇痛药效筛选。结果唐古特瑞香乙醚部位(0.44 g/kg)可明显减少冰醋酸所致的小鼠扭体次数(P<0.01)及冰醋酸所致小鼠毛细血管通透性增加(P<0.01)和抑制二甲苯所致小鼠耳肿胀(P<0.05)。结论唐古特瑞香乙醚部位为抗炎镇痛的药效部位,为唐古特瑞香开发研究提供科学依据。     

芫花对P-糖蛋白底物罗丹明123肠内转运和吸收的影响

 目的 研究芫花水煎液对P-糖蛋白(P-gp)的底物罗丹明123(R123)在空肠内转运和吸收的影响，探讨芫花对肠黏膜P-gp的调控作用。方法 大鼠分组制模，利用体外扩散池法(Ussing chamber)评价芫花对R123和荧光素钠(CF)经空肠黏膜透过性的影响；利用在体实验测定R123和CF在血浆中药物浓度的变化和各种药动学参数。结果 一方面，0.5 g·mL-1的芫花水煎液降低了R123分泌方向的转运，同时提高了吸收方向的转运，而且对CF吸收和分泌方向的转运都有抑制作用；另一方面，在体实验芫花组R123的生物利用度和曲线下面积都显著大于对照组，CF则与对照组无明显差异。结论 实验表明，芫花某些化学成分抑制P-gp，另一些成分可能使细胞之间紧密连接加强，引起旁细胞途径药物CF渗透的降低。因此芫花是肠黏膜P-gp的一种抑制剂，但是哪种化学成分发挥作用有待于进一步研究。     

Genoprotective and neuroprotective effects of Daphne gnidium leaf methanol extract,tested on male mice

Methanol extract of Daphne gnidium leaves was assessed for its antigenotoxic and neuroprotective effects through antioxidant and antibutyrylcholinesterase activities. Antigenotoxic activity was evaluated against methyl methanesulfonate injected intraperitoneally to mice, using the comet assay. The protective effect of D. gnidium reached 99.12%, at the lowest tested dose (44?mg/kg b.w.) in kidney cells, and 92.16% at the dose of 88?mg/kg b.w. in blood cells. The extract was dissolved in water and administrated to mice by intraperitoneal injection. Antioxidant activity was tested against DPPH radicals. It reached a maximum of 74.52% with an IC₅₀ value of 45?µg/ml. Anticholinesterase activity was determined against butyrylcholinesterase, an enzyme linked to Alzheimer disease. The extract exhibited antibutyrylcholinestrase effect with an inhibition percentage of 35.82% at the lowest tested dose (44?mg/kg b.w.).     

Second laparotomy for proximal colon cancer. Sites of recurrence and implications for adjuvant therapy

Sixty-four patients with primary adenocarcinomas of the colon proximal to the peritoneal reflection underwent exploratory laparotomy at the time of first recurrence of their cancers. Local recurrence, retroperitoneal lymph node metastases, parenchymal liver metastases, and diffuse peritoneal seeding were identified as frequent components of the overall pattern of recurrence. The significance of these findings has been discussed herein, relative to refinements of operative technique and selection of appropriate adjuvant therapy.