板蓝根脂溶性成分的GC-MS分析

目的:分析、鉴定板蓝根的脂溶性成分。方法:采用石油醚萃取板蓝根的脂溶性成分,进行甲酯化处理后用气相色谱-质谱联用技术分离和鉴定其组成和含量。结果:从石油醚萃取部位共鉴定出42种脂溶性成分,其中脂肪酸甲酯化产物占36.72%(饱和脂肪酸甲酯为8.67%、不饱和脂肪酸甲酯为28.05%);有35种成分在板蓝根脂溶性成分的研究中未见报道。结论:板蓝根脂溶性成分可为其综合利用以及开发抗氧化方面应用提供参考。     

板蓝根化学成分研究

目的从板蓝根中分离化学成分。方法板蓝根95%乙醇提取物经硅胶吸附,不同溶剂洗脱部分经硅胶柱层析反复分离纯化,测定所得化合物的理化常数和波谱数据,鉴定其化学结构。结果从氯仿及正丁醇部位分离得5个化合物,分别鉴定为4—(1,2,3—三羟基丙基)—2,6—二甲氧基苯—1—O—β—D—葡萄糖苷(Ⅰ)、香草醛(Ⅱ)、甘露醇(Ⅲ)、琥珀酸(Ⅳ)、苯甲酸(Ⅴ)。结论化合物Ⅰ、Ⅱ、Ⅲ为首次从板蓝根药材中分离,具有一定的参考价值。     

板蓝根化学成分的研究(Ⅰ)

目的 从板蓝根中分离化学成分.方法 将板蓝根的95%乙醇提取物经硅胶吸附,不同溶剂洗脱部分经硅胶柱层析分离纯化,测定所得化合物的理化常数和波谱数据,鉴定化学结构.结果 从石油醚部位分得5个化合物:β-谷甾醇十二烷酸酯(Ⅰ)、十七烷酸(Ⅱ)、棕榈酸(Ⅲ)、豆甾醇(Ⅳ)、β-谷甾醇(Ⅴ).结论 化合物Ⅰ、Ⅱ、Ⅳ为首次从板蓝根药材中分离.     

几丁糖预防CO2激光声带切除术后声带粘连短期观察的实验研究

Objective To observe the effect of immediate topical application of chitosan on preventing anterior giottic stenosis (AGS) after microsurgical resection of both vocal fold with CO<,2> laser , including the anterior commissure, in a canine model. Methods Sixteen canine larynges were injured by microresecting procedure of both vocal folds with CO<,2> laser. The dogs were randomly divided into two groups, chitosan group and control group. The chitosan and isotonic sodium chloride solution(control) were used for 5 minutes immediately after surgery. One week after the initial surgery, three dogs in each group were randomly selected , ultrastructure of fibroblast were examined with transmission electronic microscope and expression of basic fibroblast growth factor(bFGF) and traansforming growth factor betal (TGF-β1) were evaluated by enzyme-linked immunosorbent assay(ELISA). Three weeks after surgery, the rest dogs' glottic web were lysed and repeatedly treated with chitosan and isotonic sodium chloride solution respectively. The glottic wound healing and AGS formation were examined every week, and all larynges were harvested and examined histologically six weeks after the initial surgery. Results Transmission electronic microscope examination of the ultrasmcture of fibroblast indicated that chitosan inhibited the proliferation of fibroblast. Chitosan increased the expression of bFGF and TGF-β1, and bFGF and TGF-β1 in chitosan group, which was significantly higher than that in control group (z = -2.887 and -2.005, P =0.002 and 0.041). Chitosan decreased the extent of AGS formation. Three weeks after the surgery, the AGS lesion in the control group affected mean 49% of the length of the vocal folds from the anterior commissure to the vocal process, while chitosan group affected mean 7%, which was significantly less than the extent of web formation in the control group, (z = - 2. 619, P = 0. 008). The grade of collagen content in chitosan group was significantly lower than that in control group (P = 0. 003). Conclusion Chitosan is effective in preventing AGS after CO<,2> laser cordectomy.     

小檗碱通过激活自噬诱导caspase依赖性凋亡发挥抗结直肠癌作用

  目的  探究小檗碱通过自噬依赖性凋亡发挥抗结直肠功效的潜在作用机制。  方法  使用HCT116细胞和移植瘤模型小鼠进行小檗碱抗肿瘤药效和可能作用机制的研究。体外研究中, 通过利用CCK8实验评价小檗碱对结直肠癌细胞活力的影响, 使用细胞克隆实验评价小檗碱对结直肠癌细胞增殖的影响, 同时借助流式细胞术和TUNEL染色法对小檗碱诱导结直肠癌细胞凋亡作用进行研究。透射电镜和mCherry-GFP-LC3B腺病毒转染细胞用于检测细胞内自噬流变化。通过对小鼠组织进行HE染色、免疫组化染色和TUNEL染色, 评价小檗碱体内抗结直肠癌作用。  结果  体外研究结果表明, 小檗碱能够抑制结直肠癌细胞活力, 诱导细胞凋亡, 同时提高细胞内LC3B水平。使用自噬抑制剂3-MA、CQ和BafA1进行干预, 自噬抑制剂能够促进HCT116细胞生长, 抵消小檗碱诱导的结直肠癌细胞凋亡作用。雷帕霉素增强小檗碱上调HCT116细胞中LC3B水平的作用, 同时Z-VAD-FMK或ATG siRNA能够废除小檗碱诱导结直肠癌细胞凋亡的作用。在体研究结果表明, 小檗碱能够降低肿瘤体积和重量, 引起肿瘤组织发生凋亡。进一步的体内作用机制研究结果表明, 小檗碱显著抑制p-mTOR表达, 同时显著上调ATG5、ATG7、cleaved-caspase3和cleaved-caspase8表达。  结论  小檗碱能够通过诱导结直肠癌细胞发生自噬, 促进其发生caspase依赖性凋亡, 进而抑制结直肠癌细胞增殖。      

几丁糖预防CO2激光声带切除术后声带粘连短期观察的实验研究

Objective To observe the effect of immediate topical application of chitosan on preventing anterior giottic stenosis (AGS) after microsurgical resection of both vocal fold with CO<,2> laser , including the anterior commissure, in a canine model. Methods Sixteen canine larynges were injured by microresecting procedure of both vocal folds with CO<,2> laser. The dogs were randomly divided into two groups, chitosan group and control group. The chitosan and isotonic sodium chloride solution(control) were used for 5 minutes immediately after surgery. One week after the initial surgery, three dogs in each group were randomly selected , ultrastructure of fibroblast were examined with transmission electronic microscope and expression of basic fibroblast growth factor(bFGF) and traansforming growth factor betal (TGF-β1) were evaluated by enzyme-linked immunosorbent assay(ELISA). Three weeks after surgery, the rest dogs' glottic web were lysed and repeatedly treated with chitosan and isotonic sodium chloride solution respectively. The glottic wound healing and AGS formation were examined every week, and all larynges were harvested and examined histologically six weeks after the initial surgery. Results Transmission electronic microscope examination of the ultrasmcture of fibroblast indicated that chitosan inhibited the proliferation of fibroblast. Chitosan increased the expression of bFGF and TGF-β1, and bFGF and TGF-β1 in chitosan group, which was significantly higher than that in control group (z = -2.887 and -2.005, P =0.002 and 0.041). Chitosan decreased the extent of AGS formation. Three weeks after the surgery, the AGS lesion in the control group affected mean 49% of the length of the vocal folds from the anterior commissure to the vocal process, while chitosan group affected mean 7%, which was significantly less than the extent of web formation in the control group, (z = - 2. 619, P = 0. 008). The grade of collagen content in chitosan group was significantly lower than that in control group (P = 0. 003). Conclusion Chitosan is effective in preventing AGS after CO<,2> laser cordectomy.     

板蓝根水溶性化学成分的研究

目的提取、分离板蓝根水溶性化学成分。方法板蓝根水提取液经D101大孔树脂吸附,不同浓度乙醇洗脱部分经硅胶柱层析反复分离、纯化,测定所得化合物的理化常数和波谱数据,鉴定其化学结构。结果从板蓝根水提取液中暂时分得3个化合物,分别鉴定为丁香苷(Ⅰ)、吲哚—3—乙腈—6—O—β—D—葡萄糖苷(Ⅱ)、(+)—异落叶松树脂醇(Ⅲ)。结论化合物Ⅰ、Ⅱ系首次从板蓝根中分离。     

更正

腺样体肥大是小儿常见疾患，为小儿阻塞性睡眠呼吸暂停低通气综合征的主要原因。我科自2001年7月～2005年1月，应用小儿鼻内镜及切割器经口、鼻联合进路行腺样体切除术85例，取得良好效果，总结如下。     

UPLC-MS/MS法测定大鼠血浆中紫草氰苷、格列风内酯浓度

目的 建立使用UPLC-MS/MS法测定大鼠血浆中紫草氰苷、格列风内酯浓度的方法,并研究其药物代谢动力学特征。方法 采用Aglient C₁₈色谱柱(100 mm×2.1 mm,1.8 μm),流动相为乙腈(A)-0.1%甲酸水溶液(B),梯度洗脱(0~1 min,5%~20%B;1~2 min,20%~60%B;2~6 min,60%~70%B;6~6.5 min,70%~5%B;6.5~7.5 min,5%B),流速0.3 mL/min,柱温35 ℃。大鼠灌胃给药量为4 g/(kg·d),使用液质联用色谱法测定给药后不同时间点紫草氰苷和格列风内酯的血药浓度,通过DAS2.0软件模拟计算,得出相应的药物代谢动力学参数。结果 在大鼠血浆中,紫草氰苷在0.701 9~140.4 ng/mL,格列风内酯在1.120~240.4 ng/mL内线性关系良好,两者日内、日间精密度和准确度RSD均小于15%,回收率在87.22%~107.75%之间。紫草氰苷和格列风内酯在体内吸收均较迅速,0.5 h达到最高血药浓度,并且消除速率较快,24 h基本消除。结论 经方法学检验,此方法准确、稳定,可用于紫草氰苷和格列风内酯的药物代谢动力学研究。     

缺氧微环境下仙连解毒方抑制结直肠癌细胞增殖的作用及机制

目的 观察缺氧微环境下仙连解毒方对含溴结构域蛋白4（Brd4）诱导的核转录因子-κB（NF-κB）信号通路激活的影响,探讨其抑制结直肠癌细胞HT-29增殖的作用机制。方法 于缺氧培养箱或常氧培养箱中培养人结直肠癌细胞HT-29,给予仙连解毒方（0.8、1、1.2、1.6、3.2、6.4、12.8 g·L^-1）干预细胞48 h,采用噻唑蓝（MTT）比色法检测细胞活力;采用线粒体膜电位荧光探针（JC-1）检测仙连解毒方（1.25、2.5、5 g·L^-1）对细胞线粒体膜电位的影响,采用流式细胞术检测结直肠癌细胞HT-29凋亡情况;细胞克隆形成实验及5-乙炔基-2''-脱氧尿苷（EDU）法检测结直肠癌细胞HT-29细胞增殖能力;蛋白免疫印迹法（Western blot）检测Brd4及其下游蛋白如c-核蛋白类基因（c-Myc）、六亚甲基双乙酰胺诱导蛋白1（HEXIM1）的表达水平,同时检测仙连解毒方对NF-κB信号通路相关蛋白的影响。结果 与空白组比较,仙连解毒方（0.8、1、1.2、1.6、3.2、6.4、12.8 g·L^-1）组均能抑制结直肠癌细胞HT-29细胞活力（P<0.05,P<0.01）,且缺氧培养下细胞半数抑制浓度（IC₅₀）>常氧培养组。与空白组比较,仙连解毒方（1.25、2.5、5 g·L^-1）组细胞线粒体膜电位明显下降、细胞凋亡增多（P<0.05,P<0.01）。与空白组比较,仙连解毒方（1.25、2.5、5 g·L^-1）组细胞克隆数减少,EDU阳性细胞数减少（P<0.05,P<0.01）。Western blot结果示,与空白组比较,仙连解毒方（1.25、2.5、5 g·L^-1）组细胞内Brd4、c-Myc蛋白表达量均有不同程度的下降,HEXIM1表达升高（P<0.05,P<0.01）;磷酸化（p）-NF-κB p65、磷酸化NF-κB抑制蛋白α（p-IκBα）蛋白表达下调（P<0.05,P<0.01）。结论 缺氧微环境下仙连解毒方可抑制Brd4调控的结直肠癌细胞HT-29增殖,抑制NF-κB信号通路的激活可能是其机制之一。