目的建立桃仁多指标成分高效液相色谱法(high performance liquid chromatography,HPLC)定量分析方法,联合化学计量学、加权逼近理想解排序(technique for order preference by similarity to ideal solution,TOPSIS)法与灰色关联度分析...目的建立桃仁多指标成分高效液相色谱法(high performance liquid chromatography,HPLC)定量分析方法,联合化学计量学、加权逼近理想解排序(technique for order preference by similarity to ideal solution,TOPSIS)法与灰色关联度分析(grey correlation analysis,GRA)融合模型,科学评价不同产地桃仁的质量。方法采用Develosil ODS-UG-5 C_(18)色谱柱,以乙腈-体积分数0.2%磷酸溶液为流动相进行梯度洗脱,检测波长为210 nm(用于检测苦杏仁苷、野黑樱苷、β-谷甾醇、岩藻甾醇)和360 nm(用于检测儿茶素、山柰酚-3-O-芸香糖苷、槲皮素、柚皮素、山柰酚),柱温为30℃,建立HPLC多指标定量方法。采用外标法测定18批不同产地桃仁中上述9种成分的含量;参照《中华人民共和国药典》(2020年版)四部通则,测定酸值、羰基值、总灰分、酸不溶性灰分和醇浸出物含量。对检测数据进行化学计量学分析,建立加权TOPSIS与GRA融合评价模型。结果建立的HPLC定量方法在检测中,各成分线性关系良好,方法的重复性、准确性及仪器精密度均符合要求;体积分数70%甲醇回流提取制备的桃仁供试品溶液在24 h内稳定性良好。外标法测定结果显示,不同产地桃仁样品质量存在显著差异。化学计量学分析结果显示,18批桃仁可聚为3类,苦杏仁苷、野黑樱苷、槲皮素、β-谷甾醇和山柰酚是影响桃仁质量的关键差异性标志物。融合模型评价结果显示,18批样品的综合相对贴近度为0.2845~0.6835。结论建立的HPLC多指标定量方法操作便捷、稳定可靠,适用于桃仁多指标成分的同步测定;化学计量学联合加权TOPSIS与GRA融合模型,为不同产地桃仁的质量综合评价提供了技术支撑。展开更多
Xiao-Xu-Ming decoction has been widely used to treat stroke and sequelae of stroke. We have previously shown that the active fractions of Xiao-Xu-Ming decoction attenuate cerebral ischemic injury. However, the global ...Xiao-Xu-Ming decoction has been widely used to treat stroke and sequelae of stroke. We have previously shown that the active fractions of Xiao-Xu-Ming decoction attenuate cerebral ischemic injury. However, the global protein profile and signaling conduction pathways regulated by Xiao-Xu-Ming decoction are still unclear. This study established a two-vessel occlusion rat model by bilateral common carotid artery occlusion. Rats were intragastrically administered 50 or 150 mg/kg Xiao-Xu-Ming decoction for 4 consecutive weeks. Learning and memory abilities were measured with Morris water maze. Motor ability was detected with prehensile test. Coordination ability was examined using the inclined screen test. Neuronal plasticity was observed by immunofluorescent staining. Differentially expressed proteins of rat hippocampus were analyzed by label-free quantitative proteomics. Real time-polymerase chain reaction and western blot assay were used to identify the changes in proteins. Results showed that Xiao-Xu-Ming decoction dramatically alleviated learning and memory deficits, and motor and coordination dysfunction, and increased the expression of microtubule-associated protein 2. Xiao-Xu-Ming decoction extract remarkably decreased 13 upregulated proteins and increased 39 downregulated proteins. The regulated proteins were mainly involved in oxidation reduction process, intracellular signaling cascade process, and protein catabolic process. The signaling pathways were mainly involved in ubiquitin mediated proteolysis and the phosphatidylinositol signaling system. Furthermore, there was an interaction among Rab2 a, Ptpn1, Ppm1 e, Cdk18, Gorasp2, Eps15, Capza2, Syngap1 and Mt-nd1. Protein analyses confirmed the changes in expression of MTND1. The current findings provide new insights into the molecular mechanisms of Xiao-Xu-Ming decoction extract's effects on chronic cerebral hypoperfusion.展开更多
In the present study, we aimed to investigate the effects of Xiao-Xu-Ming decoction extract(XXM) on lipopolysaccaride(LPS)-induced neuroinflammation in vitro and in vivo. In vitro, the microglia BV2 cells were treated...In the present study, we aimed to investigate the effects of Xiao-Xu-Ming decoction extract(XXM) on lipopolysaccaride(LPS)-induced neuroinflammation in vitro and in vivo. In vitro, the microglia BV2 cells were treated with 200 ng/mL LPS for 24 h to induce inflammatory responses. In vivo, mice were treated with 5 mg/kg LPS to induce inflammatory responses. The NO level was determined by Griess Reagents. The levels of IL-1β, IL-6, TNF-α and MCP-1 were determined by ELISA. The expressions of Iba-1, TLR4 and MyD88 at the protein levels were determined by Western blotting analysis. The mRNA levels of TLR4 and MyD88 were determined by real-time PCR. In vitro, XXM significantly reduced the levels of various pro-inflammatory factors, including NO, IL-1β, IL-6 and TNF-α, induced by LPS in the supernatant of BV2 cells and suppressed expressions of inflammatory proteins TLR4 and MyD88 induced by LPS in BV2 cells. In vivo, XXM significantly inhibited microglia activation, attenuated LPS-induced inflammatory factors and chemokine production, such as IL-1β, IL-6, TNF-α and MCP-1, and inhibited the expressions of inflammatory proteins including TLR4 and MyD88, in the cortex of LPS-induced mice. Our findings suggested that XXM could attenuate LPS-induced neuroinflammation via down-regulating TLR4/MyD88 signaling pathway.展开更多
OBJECTIVE Vascular dementia(VD) refers to a progressive decline in memory and cognitive function caused by chronic cerebral ischemia.2-Vessels occlusion(2-VO) has been widely used as a model of VD.Xiao-Xu-Ming decocti...OBJECTIVE Vascular dementia(VD) refers to a progressive decline in memory and cognitive function caused by chronic cerebral ischemia.2-Vessels occlusion(2-VO) has been widely used as a model of VD.Xiao-Xu-Ming decoction,a well-known traditional Chinese medicine prescrip.tion,has been widely used to treat stroke and sequelae of stroke.The present study was to investigate the mechanism of Xiao-Xu-Ming decoction(XXM) against chronic cerebral ischemia injury in rats.METHODS After XXM treatment,rats were performed a memory testing with Morris water maze and motor ability testing using prehensile test and inclined screen test.Neuronal plasticity was observed by immunofluorescent staining with MAP2 antibody.Differentially expressed proteins of rat hippocampus were analyzed by Label-free quantitative proteomics.RESULTS XXM significantly alleviated 2-VOinduced learning and memory deficits,motor ability dysfunction,and neuronal plasticity injury in rats.The mechanism might be involved in up-regulation of 39 proteins and down-regulation of 13 proteins in the hippocampus of rats after XXM treatment vs 2-VO group rats.Gene ontology and pathway analysis showed that the regulated proteins are mainly involved in oxidation reduction process,intracellular signaling cascade process,and protein catabolic process,etc.The signal pathways are mainly involved in ubiquitin mediated proteolysis and phosphatidylinositol signaling system.CONCLUSION Current findings provide new insights into the molecular mechanisms of XXM on chronic cerebral ischemia.展开更多
目的探讨肺组织硅藻检验在法医学溺死诊断中的应用价值。方法将实验动物随机分为溺死组、死后入水组和陆地死亡组,采用微波消解-真空抽滤-扫描电镜联用的硅藻检验方法对肺组织和溺液硅藻进行定量分析,记录肺组织与溺液硅藻含量的比值(ra...目的探讨肺组织硅藻检验在法医学溺死诊断中的应用价值。方法将实验动物随机分为溺死组、死后入水组和陆地死亡组,采用微波消解-真空抽滤-扫描电镜联用的硅藻检验方法对肺组织和溺液硅藻进行定量分析,记录肺组织与溺液硅藻含量的比值(ratio of content of diatoms in lung tissue and drowning fluid,C_L/C_D)。结果溺死组实验兔的C_L/C_D值(5.82±3.50)高于死后入水组(0.47±0.35);溺死组实验猪肺叶不同部位的C_L/C_D值均高于死后入水组;在海水、咸淡水交界、河道淡水及湖泊淡水中,溺死组实验猪的C_L/C_D值均高于死后入水组(P<0.05)。动物实验中,C_L/C_D值>1.6的案例均来自溺死组。结论C_L/C_D值是溺死诊断中有较好应用前景的指标。展开更多
文摘目的建立桃仁多指标成分高效液相色谱法(high performance liquid chromatography,HPLC)定量分析方法,联合化学计量学、加权逼近理想解排序(technique for order preference by similarity to ideal solution,TOPSIS)法与灰色关联度分析(grey correlation analysis,GRA)融合模型,科学评价不同产地桃仁的质量。方法采用Develosil ODS-UG-5 C_(18)色谱柱,以乙腈-体积分数0.2%磷酸溶液为流动相进行梯度洗脱,检测波长为210 nm(用于检测苦杏仁苷、野黑樱苷、β-谷甾醇、岩藻甾醇)和360 nm(用于检测儿茶素、山柰酚-3-O-芸香糖苷、槲皮素、柚皮素、山柰酚),柱温为30℃,建立HPLC多指标定量方法。采用外标法测定18批不同产地桃仁中上述9种成分的含量;参照《中华人民共和国药典》(2020年版)四部通则,测定酸值、羰基值、总灰分、酸不溶性灰分和醇浸出物含量。对检测数据进行化学计量学分析,建立加权TOPSIS与GRA融合评价模型。结果建立的HPLC定量方法在检测中,各成分线性关系良好,方法的重复性、准确性及仪器精密度均符合要求;体积分数70%甲醇回流提取制备的桃仁供试品溶液在24 h内稳定性良好。外标法测定结果显示,不同产地桃仁样品质量存在显著差异。化学计量学分析结果显示,18批桃仁可聚为3类,苦杏仁苷、野黑樱苷、槲皮素、β-谷甾醇和山柰酚是影响桃仁质量的关键差异性标志物。融合模型评价结果显示,18批样品的综合相对贴近度为0.2845~0.6835。结论建立的HPLC多指标定量方法操作便捷、稳定可靠,适用于桃仁多指标成分的同步测定;化学计量学联合加权TOPSIS与GRA融合模型,为不同产地桃仁的质量综合评价提供了技术支撑。
基金supported in part by the National Natural Science Foundation of China,No.81473383(to YHW)the Significant New-Drugs Creation of Science and Technology Major Projects in China,No.2018ZX09711001-003-019(to YHW)the Innovation Fund for Graduate of Beijing Union Medical College of China,No.2017-1007-02(to XC)
文摘Xiao-Xu-Ming decoction has been widely used to treat stroke and sequelae of stroke. We have previously shown that the active fractions of Xiao-Xu-Ming decoction attenuate cerebral ischemic injury. However, the global protein profile and signaling conduction pathways regulated by Xiao-Xu-Ming decoction are still unclear. This study established a two-vessel occlusion rat model by bilateral common carotid artery occlusion. Rats were intragastrically administered 50 or 150 mg/kg Xiao-Xu-Ming decoction for 4 consecutive weeks. Learning and memory abilities were measured with Morris water maze. Motor ability was detected with prehensile test. Coordination ability was examined using the inclined screen test. Neuronal plasticity was observed by immunofluorescent staining. Differentially expressed proteins of rat hippocampus were analyzed by label-free quantitative proteomics. Real time-polymerase chain reaction and western blot assay were used to identify the changes in proteins. Results showed that Xiao-Xu-Ming decoction dramatically alleviated learning and memory deficits, and motor and coordination dysfunction, and increased the expression of microtubule-associated protein 2. Xiao-Xu-Ming decoction extract remarkably decreased 13 upregulated proteins and increased 39 downregulated proteins. The regulated proteins were mainly involved in oxidation reduction process, intracellular signaling cascade process, and protein catabolic process. The signaling pathways were mainly involved in ubiquitin mediated proteolysis and the phosphatidylinositol signaling system. Furthermore, there was an interaction among Rab2 a, Ptpn1, Ppm1 e, Cdk18, Gorasp2, Eps15, Capza2, Syngap1 and Mt-nd1. Protein analyses confirmed the changes in expression of MTND1. The current findings provide new insights into the molecular mechanisms of Xiao-Xu-Ming decoction extract's effects on chronic cerebral hypoperfusion.
基金The National Natural Science Foundation of China(Grant No.81473383)the Innovation Fund for Graduate of Beijing Union Medical College(Grant No.2017-1007-02)+1 种基金the Drug Innovation Major Project(Grant No.2018ZX09711001-003-019)the Medical and Health Innovation Project of Chinese Academy of Medical Sciences(Grant No.2016-I2M-3-007,2018-1007-04)
文摘In the present study, we aimed to investigate the effects of Xiao-Xu-Ming decoction extract(XXM) on lipopolysaccaride(LPS)-induced neuroinflammation in vitro and in vivo. In vitro, the microglia BV2 cells were treated with 200 ng/mL LPS for 24 h to induce inflammatory responses. In vivo, mice were treated with 5 mg/kg LPS to induce inflammatory responses. The NO level was determined by Griess Reagents. The levels of IL-1β, IL-6, TNF-α and MCP-1 were determined by ELISA. The expressions of Iba-1, TLR4 and MyD88 at the protein levels were determined by Western blotting analysis. The mRNA levels of TLR4 and MyD88 were determined by real-time PCR. In vitro, XXM significantly reduced the levels of various pro-inflammatory factors, including NO, IL-1β, IL-6 and TNF-α, induced by LPS in the supernatant of BV2 cells and suppressed expressions of inflammatory proteins TLR4 and MyD88 induced by LPS in BV2 cells. In vivo, XXM significantly inhibited microglia activation, attenuated LPS-induced inflammatory factors and chemokine production, such as IL-1β, IL-6, TNF-α and MCP-1, and inhibited the expressions of inflammatory proteins including TLR4 and MyD88, in the cortex of LPS-induced mice. Our findings suggested that XXM could attenuate LPS-induced neuroinflammation via down-regulating TLR4/MyD88 signaling pathway.
基金supported by National Natural Science Foundation of China(81473383) National Science and Technology Major Project of China(2018ZX09711001-003-019)+1 种基金 CAMS Innovation Fund for Medical Sciences(2016-I2M-3-007) Innovation Fund for Graduate of Beijing Union M
文摘OBJECTIVE Vascular dementia(VD) refers to a progressive decline in memory and cognitive function caused by chronic cerebral ischemia.2-Vessels occlusion(2-VO) has been widely used as a model of VD.Xiao-Xu-Ming decoction,a well-known traditional Chinese medicine prescrip.tion,has been widely used to treat stroke and sequelae of stroke.The present study was to investigate the mechanism of Xiao-Xu-Ming decoction(XXM) against chronic cerebral ischemia injury in rats.METHODS After XXM treatment,rats were performed a memory testing with Morris water maze and motor ability testing using prehensile test and inclined screen test.Neuronal plasticity was observed by immunofluorescent staining with MAP2 antibody.Differentially expressed proteins of rat hippocampus were analyzed by Label-free quantitative proteomics.RESULTS XXM significantly alleviated 2-VOinduced learning and memory deficits,motor ability dysfunction,and neuronal plasticity injury in rats.The mechanism might be involved in up-regulation of 39 proteins and down-regulation of 13 proteins in the hippocampus of rats after XXM treatment vs 2-VO group rats.Gene ontology and pathway analysis showed that the regulated proteins are mainly involved in oxidation reduction process,intracellular signaling cascade process,and protein catabolic process,etc.The signal pathways are mainly involved in ubiquitin mediated proteolysis and phosphatidylinositol signaling system.CONCLUSION Current findings provide new insights into the molecular mechanisms of XXM on chronic cerebral ischemia.
文摘目的探讨肺组织硅藻检验在法医学溺死诊断中的应用价值。方法将实验动物随机分为溺死组、死后入水组和陆地死亡组,采用微波消解-真空抽滤-扫描电镜联用的硅藻检验方法对肺组织和溺液硅藻进行定量分析,记录肺组织与溺液硅藻含量的比值(ratio of content of diatoms in lung tissue and drowning fluid,C_L/C_D)。结果溺死组实验兔的C_L/C_D值(5.82±3.50)高于死后入水组(0.47±0.35);溺死组实验猪肺叶不同部位的C_L/C_D值均高于死后入水组;在海水、咸淡水交界、河道淡水及湖泊淡水中,溺死组实验猪的C_L/C_D值均高于死后入水组(P<0.05)。动物实验中,C_L/C_D值>1.6的案例均来自溺死组。结论C_L/C_D值是溺死诊断中有较好应用前景的指标。
