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OBJECTIVE： To establish a method for simultaneous determination of 10 isoflavones in Belamcanda chinensis， and to evaluate the differences of active ingredient content of B. chinensis from different areas. METHODS： UPLC method was adopted. The determination was performed on Waters ACQUITY UPLC BEH C18 column with mobile phase consisted of 0.5 ％ methyl-β-cyclodextrin and 0.1％ phosphate as water phase， acetonitrile as organic phase （gradient elution） at the flow rate of 0.2 mL/min. The column temperature was set at 35 ℃， and the detection wavelength was set at 265 nm. The sample size was 2 μL， and analysis time was 20 min. The contents of 10 isoflavones in 26 samples from 8 provinces， including tectoridin， iristectorin A， iristectorin B， iridin， tectorigenin， iristectorigenin B， iristectorigenin A， irigenin， irisflorentin， dichotomitin， were determined. RESULTS： The linear ranges of tectoridin， iristectorin A， iristectorin B， iridin， tectorigenin， iristectorigenin B， iristectorigenin A， irigenin， irisflorentin， dichotomitin were 8.569 5-342.78， 0.643-25.72， 1.119 8-44.79， 2.187 8-87.51， 0.770 3-30.81， 0.421 3- 16.85， 0.288 5-11.54， 1.795 3-71.81， 0.560 8-22.43， 0.086-3.44 μg/mL（all r≥0.999 6）. The limits of quantitation were 0.015， 0.102， 0.096， 0.013， 0.036， 0.088， 0.102， 0.019， 0.067， 0.092 μg/mL. RSDs of precision， stability（24 h）and reproducibility tests were lower than 2.00％ （n＝6）. The recoveries ranged 95.30％-103.30％ （all RSD≤2.33％， n＝6）. Among 26 samples of B. chinensis， the content of tectoridin was the highest （3.66％-57.79％）， and the content of dichotomitin was the lowest （0.09％- 0.59％）， the contents of irisflorentin were 0.29-2.80 mg/g. The contents of isoflavones in B. chinensis from different areas were different greatly. CONCLUSIONS： The established method is sensitive， with short analysis time and good repeatability， and can be used to determine the content of 10 isoflavones and evaluate the content difference of each component.
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OBJECTIVE:To study the effects of Dianxianqing granule on the interleukin(IL-6)content and glial fibrillary acid-ic protein(GFAP),ionized calcium linker molecules 1(Iba-1)expressions in hippocampus tissue of rats with kainate-induced epi-lepsy,and explore its mechanism of preventing and treating epilepsy. METHODS:Rats were randomly divided into sham operation group (distilled water),model group (distilled water),phenytoin group (0.03 g/kg,positive control) and Dianxianqing granule low-dose,medium-dose,high-dose groups(4.74,9.47,18.94 g/kg,calculated by crude drug),20 in each group. Rats were intra-gastrically administrated once a day,for 7 d. After 1 h of last administration,except for sham operation group,rats in other groups received single injection of kainite in hippocampus CA1 of left side to induce the epilepsy model. Behavioral changes and death of rats were observed. After 24 h of modeling,enzyme-linked immunosorbent method was used to detect the IL-6 content in hippocam-pus tissue of rats,Nissl staining was used to count the hippocampus neurons,and immunohistochemistry was used to detect the GFAP,Iba-1 expressions in hippocampus tissue of rats. RESULTS:Compared with sham operation group,rats in model group had obvious epilepsy symptoms after modeling,and parts of rats died;IL-6 content and number of neurons in hippocampus tissue were obviously decreased (P<0.01), while GFAP, Iba-1 expressions were obviously enhanced (P<0.01). Compared with model group,epilepsy symptoms and death in each administration group had improved,while IL-6 content in hippocampus tissue were in-creased to varying degrees,with no statistical significance (P>0.05). The numbers of neurons in phenytoin group,Dianxianqinggranule medium-dose, high-dose groups were obviously in-creased(P<0.01);GFAP expression was obviously decreased (P<0.01). Iba-1 expressions in hippocampus tissue in phenyt-oin group,Dianxianqing granule high-dose group were obvi-ously decreased (P<0.01). CONCLUSIONS:Dianxianqing granule can play the role in preventing and treating epilepsy by inhibiting GFAP,Iba-1 expressions in hippocampus tissue and in-creasing the number of neurons in hippocampus tissue.
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Translational medicine means that to establish a bridge between fundamental research and clinical medicine through bidirectional translation path in order to promote the translation and application from fundamental research to clinical medicine.The purpose of traditional Chinese medicine (TCM) translational model is to collate and build one multidimensional TCM translational model,rationality carry out the research on TCM translation.Three paths have been confirmed,where are from literature research to clinical verification,from clinical application to clinical verification,and from experiment to clinical verification.In this research,methodology of one path mcntioned above could be established,which is the key technology of translational medicine.Take constipation,insomnia,and post-flu cough as target diseases to investigate the herbal compounds which could be qualified for clinical application based on preliminary work.This research not only provided clinical data to TCM translation,but also established one methodology of TCM translation path--"from experiment to clinical verification".
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OBJECTIVE:To study the effects of Dianxianqing granule on the interleukin(IL-6)content and glial fibrillary acid-ic protein(GFAP),ionized calcium linker molecules 1(Iba-1)expressions in hippocampus tissue of rats with kainate-induced epi-lepsy,and explore its mechanism of preventing and treating epilepsy. METHODS:Rats were randomly divided into sham operation group (distilled water),model group (distilled water),phenytoin group (0.03 g/kg,positive control) and Dianxianqing granule low-dose,medium-dose,high-dose groups(4.74,9.47,18.94 g/kg,calculated by crude drug),20 in each group. Rats were intra-gastrically administrated once a day,for 7 d. After 1 h of last administration,except for sham operation group,rats in other groups received single injection of kainite in hippocampus CA1 of left side to induce the epilepsy model. Behavioral changes and death of rats were observed. After 24 h of modeling,enzyme-linked immunosorbent method was used to detect the IL-6 content in hippocam-pus tissue of rats,Nissl staining was used to count the hippocampus neurons,and immunohistochemistry was used to detect the GFAP,Iba-1 expressions in hippocampus tissue of rats. RESULTS:Compared with sham operation group,rats in model group had obvious epilepsy symptoms after modeling,and parts of rats died;IL-6 content and number of neurons in hippocampus tissue were obviously decreased (P<0.01), while GFAP, Iba-1 expressions were obviously enhanced (P<0.01). Compared with model group,epilepsy symptoms and death in each administration group had improved,while IL-6 content in hippocampus tissue were in-creased to varying degrees,with no statistical significance (P>0.05). The numbers of neurons in phenytoin group,Dianxianqinggranule medium-dose, high-dose groups were obviously in-creased(P<0.01);GFAP expression was obviously decreased (P<0.01). Iba-1 expressions in hippocampus tissue in phenyt-oin group,Dianxianqing granule high-dose group were obvi-ously decreased (P<0.01). CONCLUSIONS:Dianxianqing granule can play the role in preventing and treating epilepsy by inhibiting GFAP,Iba-1 expressions in hippocampus tissue and in-creasing the number of neurons in hippocampus tissue.
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This study was aimed to screen out a safe and effective traditional Chinese medicine (TCM) compound prescription in insomnia treatment, through the method of uniform design and comprehensive scoring method. It provided candidate drugs to further research and development of TCM new drug. U7(76) uniform design was used to divide groups. The test was given on sleep time prolonging effect of mice injected with pentobarbital sodium. The subliminal dosage of pentobarbital hypnosis was tested on mice injected with pentobarbital sodium. The acute toxicity (LD50) was tested on mice. Comprehensive scoring method was applied to evaluate the effect to determine the optimum dose. The results showed the optimized prescription was composed of Radix Sophorae Flavescentis and Schisandra chinensis. The optimum ratio was 1:3. It was concluded that the uniform design was applied in the experimental design with 3 factors and 6 levels. Comprehensive scoring method was used to evaluate the combined effect. The optimized prescription that put benefit and risk into consideration was composed of Radix Sophorae Flavescentis and Schisandra chinensis. The best ratio was 1:3, which can be further developed.
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Objective To discuss the content determination method ofα-asarone in Dianxianqing Granules. Methods Kromasil C18 column (250 mm×4.6 mm, 5 μm) was used, with methanol and water as mobile phase of gradient elution. The flow rate was 1.0 mL/min, and the detection wavelength was 257 nm. The column temperature was 25 ℃. Results The sample solution was prepared by reflux extracted with 70% ethanol for 20 min. α-asarone had a good linearity in the ranges of 1.24×10-3-7.44×10-3μg (r=0.999 5). The method was quite good in precision, stability and repeatability, and the average recovery ofα-asarone was 98.14%, with corresponding RSD of 2.5%. Conclusion The experiment provides an accurate and fast analytical method for α-asarone determination in compound preparation of traditional Chinese medicine.
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This study was aimed to optimize Qinshuang particle best spray granulation process conditions. The or-thogonal test method was used with grain yield and baicalin transfer rate as the inspection indexes. The main in-fluence factors of Qinshuang particle spray granulation process were investigated. The results showed that the best technological parameters are atomization pressure of 1 800 Pa, inlet temperature at 90°C - 100°C, outlet temper-ature at 50 ° C - 60 ° C , and the extract relative density of 1 . 10 ( 60 ° C ) . It was concluded that the reasonable pro-cess conditions make particles with stable quality, which provide the experimental basis for industrial production.
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Objective To investigate whether no asphyxia neonates with intrauterine distress are complicated with myocardial injury and determine the sensitive biochemical diagnostic parameters. Methods A total of 89 neonates born in the First Affiliated Hospital of Sun Yat-sen University from July 2009 to December 2009 were enrolled. Fifty-three fetal distress cases with Apgar score ＞ 7 at 1 and 5 minites were enrolled in the study group; while the rest 36 healthy neonates, whose Apgar score = 10 at 1 and 5 minites, were the control group. Umbilical artery blood samples of all cases were collected for blood gas analysis and biochemical measurement. Results(1)pH(7.23±0.07) and BE [(-4.8±3.0)mmol/L] in the study group were significantly lower than pH (7.31 ±0.03) and BE [(-2.1±1.5)mmol/L] in the control group (P＜0.05).The lactic acid of study group [(5.2±2.3)mmol/L] was higher than that of the control group [(2.3±1.1)mmol/L], and the difference was significant (P＜0.01). However, there was no significant difference between the two groups in PaO2[(16.2±7.9)mm Hg(1 mm Hg=0.133 kPa) vs. (17.5±6.7)mm Hg] and PaCO2[(54.0±11.2)mm Hg vs. (48.5±5.4) mm Hg; P＞0. 05]. (2) The level of CK-MB in neonates with fetal distress[(48 ±59) U/L] was significantly higher than that of healthy neonates [(36±27)U/L]. However, no significant difference was found in CK [(194±73)U/L vs. (162±95) U/L]and BNP levels[(519±309)ng/L vs.(481±216)ng/L;P ＞ 0.05]. (3) Spearman rank correlation analysis showed that CK-MB level was negatively correlated with pH(r=-0.296, P＜0.05) and BE (r=-0.318,P＜0.05) of umbilical artery blood,while BNP level was positively correlated with umbilical lactic acid (r=0.278, P＜0.05). No correlation was found between other parameters (P＞0.05).Conclusions Intrauterine distress without neonatal asphyxia had effect on fetal myocardial injury. CK-MB can be used as a sensitive parameter for monitoring the development of myocardial injury. The severity of myocardial injury was related to fetal acidosis.