We have developed an on-line detection method using acridine orange as the fluorescence probe and applied this method to rapidly identify active compounds in herbal medicines. This on-line method was equipped with a h...We have developed an on-line detection method using acridine orange as the fluorescence probe and applied this method to rapidly identify active compounds in herbal medicines. This on-line method was equipped with a high-performance liquid chromatography tandem diode array detector, electrospray ionization-ion-trap time-of-flight mass spectrometry and DNA- acridine orange fluorescence detection （HPLC-DAD-MSn-DNA-AO-FLD）. A large amount of information could be simultaneously obtained during one run, which included HPLC fingerprint, ultraviolet spectra, total ion chromatograms, MSn data of high-resolution mass spectrometry and activity profile of each compound binding with DNA. The method also provided information on structureactivity relationships and mechanism of interaction. We used this on-line method to identify five DNA-binding activity components from Lithospermum erythrorhizon sample for the first time. The result showed that the parent nucleus of shikonin derivatives could bind with DNA. The structure-activity relationship showed that the parent nucleus of shikonin derivatives plays a major role in DNA binding, not the carboxyl group on the side chain. This simple, rapid, high precision and good stability on-line method should be useful for compound separation, structural identification and screening of DNA-binding compounds in herbal medicines.展开更多
A culture of Lithospermum erythrorhizon adsorbed on fungal cell wall polysaccharides, a novel bio-adsorbent made from fungal cell wall, has been established in this paper. Three steps were involved in this immobilizat...A culture of Lithospermum erythrorhizon adsorbed on fungal cell wall polysaccharides, a novel bio-adsorbent made from fungal cell wall, has been established in this paper. Three steps were involved in this immobilization. The first step was preparation of suspended plant cells from tightly aggregated plant cell clumps. The disassembled ratio of 0.715g·g-1 (the disassembled cells over total cells) was obtained under optimum condition for the enzymatic reaction. Then, the adsorption of plant cells onto fungal cell wall polysaccharides was conducted and the saturated capacity of 12 g cell per gram of carrier was obtained in adsorption immobilization. Finally, the culture of cells adsorbed on fungal cell wall polysaccharides was compared with that of cells entrapped in alginate or suspension cell culture. While exposed to in situ liquid paraffin extraction coupled with cell culture, the shikonin productivity of immobilized cells by adsorption was 10.67g·L-1, which was 1.8 times of that in suspension culture and 1.5 times of that entrapped in alginate.展开更多
In order to identify the potential nephrotoxic compounds in traditional Chinese medicine Lithospermum erythrorhizon,it was separated into serial fractions according to their polarities.An in vitro method was utilized ...In order to identify the potential nephrotoxic compounds in traditional Chinese medicine Lithospermum erythrorhizon,it was separated into serial fractions according to their polarities.An in vitro method was utilized to determine the nephrotoxicity of these fractions with the help of fluorescence image analysis.As a result,the primary fraction A05 and its secondary fractions C06 ＂C09 and C12 ＂C14 were found to have significant toxicity to LLC-PK1 cell line,as determined by the survive rate less than 20% after they were treated with these fractions.These potential nephrotoxic fractions were further analyzed by multistage and high resolution mass spectrometry.The main compounds in these fractions were tentatively identified to be acetylshikonin,isobutyrylshikonin,β,β＇-dimethyla-cryloylshikonin,and isovalerylshikonin,which may bring nephrotoxicity.展开更多
[Objectives] To optimize the pet dog dyeing process of plant hair dye. [Methods]Three factors affecting dyeing were selected including pH,concentration of dye and dyeing time. The L_9(3~3) orthogonal design was used t...[Objectives] To optimize the pet dog dyeing process of plant hair dye. [Methods]Three factors affecting dyeing were selected including pH,concentration of dye and dyeing time. The L_9(3~3) orthogonal design was used to evaluate the hair coloring effect and the absorbance of cleaning solution after hair was dyed. [Results] The optimum pet dog dyeing process of plant hair dye was dye concentration of50 mg/mL,dye pH of 7. 5 and dyeing time of 1 h. [Conclusions] The coloring and fixing effects of the dye and dyeing process are good,which can lay a foundation for the development and utilization of plant dyes.展开更多
Objective:To reveal GSDME-executed pyroptosis in cancer cells induced by the Chinese traditional herbal medicine plant Lithospermum erythrorhizon(L.erythrorhizon,Zi Cao)and to investigate the potential mechanism.Metho...Objective:To reveal GSDME-executed pyroptosis in cancer cells induced by the Chinese traditional herbal medicine plant Lithospermum erythrorhizon(L.erythrorhizon,Zi Cao)and to investigate the potential mechanism.Methods:L.erythrorhizon was extracted by ultrasonication in 95%ethanol,and determined using high-performance liquid chromatography(HPLC).He La,A549,SW620,HEK-293 T,THP-1,K562,Raw264.7 and MDA-MB-231 cell lines were used to investigate the morphology and mechanism of pyroptosis induced by L.erythrorhizon.The lactate dehydrogenase(LDH)release,propidium iodide(PI)/Hoechst double-staining,and pyroptosis reconstitution experiments were performed to study L.erythrorhizon-induced cell pyroptosis.Results:Compared with the death inhibitor,PI/Hoechst and LDH release experiments,we found that L.erythrorhizon induced pyroptosis.Recombination and western blot experiments confired that L.erythrorhizon induced GSDME cleavage,which drives pyroptosis.This phenomenon is conserved in several cancer cell lines that might be triggered by caspase family proteases.The mechanism of L.erythrorhizon inducing pyroptosis is widely found in tumor cells.Conclusion:Our findings not only explain how L.erythrorhizon triggers cancer cell pyroptosis,but also provide mechanistic insights to guide its clinical application in the future.展开更多
The clinical effects of dosage during the period of treatment of Lithospermum Ruderale extract-a kind of Chinese traditional herbal medicine-on medicinal abortion induced by mifepristone and misoprostol were studied. ...The clinical effects of dosage during the period of treatment of Lithospermum Ruderale extract-a kind of Chinese traditional herbal medicine-on medicinal abortion induced by mifepristone and misoprostol were studied. Lithospermum Ruderale extract was administrated 3 d before, 3 d after or 3 d before plus 3 d after the administration of misoprostol respectively. The dose of Lithospermum Ruderale extract was 50 g, 75 g or 100 g respectively. Thus 1 350 women of early pregnancy were grouped into 9 groups and observed. The results showed that the effects of Lithospermum Ruderale used 3 d before, and 3 d before plus 3 d after (6 days misoprostol were significantly better than those only used 3 d after misoprostol both for complete abortion and bleeding (P<0.05). The dosage between 50 g and 100 g made no significant difference in clinical effects. Therefore it is reasonable to use 50 g Lithospermum Ruderale before misoprostol to improve medicinal abortion.展开更多
Early pregnancies(<42d)in 600 women were terminated respectively by mifepristone and Lithospermum Ruderal with misoprostol or mifepristone with misoprostol. The results showed that the rate of complete abortion(99....Early pregnancies(<42d)in 600 women were terminated respectively by mifepristone and Lithospermum Ruderal with misoprostol or mifepristone with misoprostol. The results showed that the rate of complete abortion(99.00%)of 300 women by mifepristone and Lithospermum Ruderal with misoprostol was significantly higher than that (92. 33%)of the other 300 women by mifepristone with misoprostol(P<0.01).The average length of time of embryo removal(54.01±7.55 h)and the rate of embryo removal (82.33%)within 6 hours after administration of misoprostol of the former were significantly shorter and higher than those(56.41±9.93 h and 72.67%) of the latter(P<0.05 respectively).The average length of the bleeding time of the former(7.01±3.11d)was significantly shorter than that(8.09±5.63d)of the latter(P<0.05).The recovery of the menstruation of both was not significantly different from each other(P>0.05).In all, the medical abortion by mifepristone and Lithospermum Ruderal with misoprostol was even more successful,with less bleeding time.Therefore it is essential to make researches on the auxiliary effects of Lithospermum Ruderal on the medical abortion by mifepristone with misoprostol.展开更多
基金Peking University Comprehensive Platform for Innovative Drug Research and Development(Grant No.2009ZX--09301--010)
文摘We have developed an on-line detection method using acridine orange as the fluorescence probe and applied this method to rapidly identify active compounds in herbal medicines. This on-line method was equipped with a high-performance liquid chromatography tandem diode array detector, electrospray ionization-ion-trap time-of-flight mass spectrometry and DNA- acridine orange fluorescence detection （HPLC-DAD-MSn-DNA-AO-FLD）. A large amount of information could be simultaneously obtained during one run, which included HPLC fingerprint, ultraviolet spectra, total ion chromatograms, MSn data of high-resolution mass spectrometry and activity profile of each compound binding with DNA. The method also provided information on structureactivity relationships and mechanism of interaction. We used this on-line method to identify five DNA-binding activity components from Lithospermum erythrorhizon sample for the first time. The result showed that the parent nucleus of shikonin derivatives could bind with DNA. The structure-activity relationship showed that the parent nucleus of shikonin derivatives plays a major role in DNA binding, not the carboxyl group on the side chain. This simple, rapid, high precision and good stability on-line method should be useful for compound separation, structural identification and screening of DNA-binding compounds in herbal medicines.
基金Supported by the National Natural Science Foundation of China (No. 29606009).
文摘A culture of Lithospermum erythrorhizon adsorbed on fungal cell wall polysaccharides, a novel bio-adsorbent made from fungal cell wall, has been established in this paper. Three steps were involved in this immobilization. The first step was preparation of suspended plant cells from tightly aggregated plant cell clumps. The disassembled ratio of 0.715g·g-1 (the disassembled cells over total cells) was obtained under optimum condition for the enzymatic reaction. Then, the adsorption of plant cells onto fungal cell wall polysaccharides was conducted and the saturated capacity of 12 g cell per gram of carrier was obtained in adsorption immobilization. Finally, the culture of cells adsorbed on fungal cell wall polysaccharides was compared with that of cells entrapped in alginate or suspension cell culture. While exposed to in situ liquid paraffin extraction coupled with cell culture, the shikonin productivity of immobilized cells by adsorption was 10.67g·L-1, which was 1.8 times of that in suspension culture and 1.5 times of that entrapped in alginate.
基金Supported by the National Key Scientific and Technological Project of China(No.2009ZX09502-012)the Research Fund for the Doctoral Program of Higher Education of China(No.20090101110126)the Zhejiang Province Science and Technology Plan Project,China(No.2008C23065)
文摘In order to identify the potential nephrotoxic compounds in traditional Chinese medicine Lithospermum erythrorhizon,it was separated into serial fractions according to their polarities.An in vitro method was utilized to determine the nephrotoxicity of these fractions with the help of fluorescence image analysis.As a result,the primary fraction A05 and its secondary fractions C06 ＂C09 and C12 ＂C14 were found to have significant toxicity to LLC-PK1 cell line,as determined by the survive rate less than 20% after they were treated with these fractions.These potential nephrotoxic fractions were further analyzed by multistage and high resolution mass spectrometry.The main compounds in these fractions were tentatively identified to be acetylshikonin,isobutyrylshikonin,β,β＇-dimethyla-cryloylshikonin,and isovalerylshikonin,which may bring nephrotoxicity.
基金Supported by the National Science and Technology Foundation Project of Traditional Chinese Veterinary Medicine Resources Rescuing and Organizing(2013FY110600-03)
文摘[Objectives] To optimize the pet dog dyeing process of plant hair dye. [Methods]Three factors affecting dyeing were selected including pH,concentration of dye and dyeing time. The L_9(3~3) orthogonal design was used to evaluate the hair coloring effect and the absorbance of cleaning solution after hair was dyed. [Results] The optimum pet dog dyeing process of plant hair dye was dye concentration of50 mg/mL,dye pH of 7. 5 and dyeing time of 1 h. [Conclusions] The coloring and fixing effects of the dye and dyeing process are good,which can lay a foundation for the development and utilization of plant dyes.
基金supported by Program for the research startup fund program at Beijing University of Chinese Medicine(90011451310011)the key research fund for drug discovery in Chinese medicine at Beijing University of Chinese Medicine(1000061223740)+1 种基金the experimental technology standardization research project at Beijing University of Chinese Medicine(2021-SYJS-009)the fundamental research funds for the central universities of Beijing University of Chinese Medicine(2020-JYBZDGG-057)。
文摘Objective:To reveal GSDME-executed pyroptosis in cancer cells induced by the Chinese traditional herbal medicine plant Lithospermum erythrorhizon(L.erythrorhizon,Zi Cao)and to investigate the potential mechanism.Methods:L.erythrorhizon was extracted by ultrasonication in 95%ethanol,and determined using high-performance liquid chromatography(HPLC).He La,A549,SW620,HEK-293 T,THP-1,K562,Raw264.7 and MDA-MB-231 cell lines were used to investigate the morphology and mechanism of pyroptosis induced by L.erythrorhizon.The lactate dehydrogenase(LDH)release,propidium iodide(PI)/Hoechst double-staining,and pyroptosis reconstitution experiments were performed to study L.erythrorhizon-induced cell pyroptosis.Results:Compared with the death inhibitor,PI/Hoechst and LDH release experiments,we found that L.erythrorhizon induced pyroptosis.Recombination and western blot experiments confired that L.erythrorhizon induced GSDME cleavage,which drives pyroptosis.This phenomenon is conserved in several cancer cell lines that might be triggered by caspase family proteases.The mechanism of L.erythrorhizon inducing pyroptosis is widely found in tumor cells.Conclusion:Our findings not only explain how L.erythrorhizon triggers cancer cell pyroptosis,but also provide mechanistic insights to guide its clinical application in the future.
基金This study was funded by National Family Planning Commission of China
文摘The clinical effects of dosage during the period of treatment of Lithospermum Ruderale extract-a kind of Chinese traditional herbal medicine-on medicinal abortion induced by mifepristone and misoprostol were studied. Lithospermum Ruderale extract was administrated 3 d before, 3 d after or 3 d before plus 3 d after the administration of misoprostol respectively. The dose of Lithospermum Ruderale extract was 50 g, 75 g or 100 g respectively. Thus 1 350 women of early pregnancy were grouped into 9 groups and observed. The results showed that the effects of Lithospermum Ruderale used 3 d before, and 3 d before plus 3 d after (6 days misoprostol were significantly better than those only used 3 d after misoprostol both for complete abortion and bleeding (P<0.05). The dosage between 50 g and 100 g made no significant difference in clinical effects. Therefore it is reasonable to use 50 g Lithospermum Ruderale before misoprostol to improve medicinal abortion.
文摘Early pregnancies(<42d)in 600 women were terminated respectively by mifepristone and Lithospermum Ruderal with misoprostol or mifepristone with misoprostol. The results showed that the rate of complete abortion(99.00%)of 300 women by mifepristone and Lithospermum Ruderal with misoprostol was significantly higher than that (92. 33%)of the other 300 women by mifepristone with misoprostol(P<0.01).The average length of time of embryo removal(54.01±7.55 h)and the rate of embryo removal (82.33%)within 6 hours after administration of misoprostol of the former were significantly shorter and higher than those(56.41±9.93 h and 72.67%) of the latter(P<0.05 respectively).The average length of the bleeding time of the former(7.01±3.11d)was significantly shorter than that(8.09±5.63d)of the latter(P<0.05).The recovery of the menstruation of both was not significantly different from each other(P>0.05).In all, the medical abortion by mifepristone and Lithospermum Ruderal with misoprostol was even more successful,with less bleeding time.Therefore it is essential to make researches on the auxiliary effects of Lithospermum Ruderal on the medical abortion by mifepristone with misoprostol.
