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OBJECTIVE： To mine and analyze the formulation rules of Chinese patent medicine for cold， and to provide reference for clinical dialectical medication and R&D of new medicine for cold. METHODS： The name， dosage form， formulation， function of curing of Chinese patent medicines for cold were collected from 2015 edition of Chinese Pharmacopeia （part Ⅰ） and then input into TCM Inheritance Assistant Platform V 2.5； use frequency of TCM were counted. Apriori algorithm and association rules were used to analyze the core medicinal material combination （10％ support and 0.65 confidence）. New formulation combinations were extracted by unsupervised entropy hierarchical clustering method. RESULTS： A total of 130 kinds of Chinese patent medicine （196 Chinese patent medicine with the same prescription and different dosage forms） for treating cold were collected， including granules （47）， pills （32）， tablets （31）， mixtures （31）， etc. 264 medicinal materials were involved. The cold syndromes contained wind-heat syndrome， wind-cold syndrome， summer-dampness syndrome and Qi deficiency syndrome. Top 3 medicinal materials in the list of use frequency were Glycyrrhiza uralensis （45.38％）， Scutellaria baicalensis （32.31％） and Platycodon grandiflorus （31.54％）. There were 28 core medicinal material combinations， among which the top 3 were G. uralensis-P. grandiflorus， Mentha haplocalyx-P. grandiflorus and S. baicalensis-Forsythia suspensa. New combinations were excavated， including Nepeta cataria-P. grandiflorus-M. haplocalyx-Citrus reticulate-Folium Perillae-Citrus aurantium- Poria cocos， F. suspense-S. baicalensis-Lonicera japonica- Arctium lappa-Fermented soybean. CONCLUSIONS： This study analyzed the formulation rules of Chinese patent medicine for treating cold by using the TCM inheritance assistant platform V2.5， which can provide reference for clinical dialectical medication and R&D of new medicines for cold.
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OBJECTIVE:To optimize the extraction technology of Compound radix fici hirta granule. METHODS:Using the transfer rate of psoralen and amygdalin in extraction liquid of Compound radix fici hirta granule and extraction rate as indexes, U12(6×4×3)uniform design was used for the test,the effects of amount of adding water,extraction time,extraction times on the extraction technology were investigated,and optimized technology was verified by three pilot scale tests. RESULTS:The optimal technology was as follow as 10-fold water,extracting for 3 times,60 min each time. Under the conditions,transfer rate of pso-ralen and amygdalin and extraction rate were 82.51%(RSD=1.45%,n=3),93.69%(RSD=0.85%,n=3),18.89%(RSD=0.74%,n=3),respectively. The validated results were in the 95% confidence interval of predictive value. CONCLUSIONS:The optimized extraction technology is stable and feasible,and provides the scientific basis for the follow-up development of the prepa-ration.
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Objective To explore the effects of manumycin on U937 cells and its mechanisms.Methods Apoptosis was detected by flow cytometry using annexin V and propidium iodide (PI) after treatment with manumycin (2 μmol/L) for different time.The reactive oxygen species (ROS) was detected by flow cytometry using Reactive Oxygen Species Assay Kit.The expressions of PI3K,P-Akt and Akt were detected by Western blotting.Results Compared with the 0 h treatment,manumycin treatment resulted in apoptosis (P ＜ 0.05) and ROS generation (P ＜ 0.05) gradually in U937 cell lines.Furthermore,manumycin also inhibited the activation of phosphatidylinositol-3 kinase (PI3K)/Akt pathway.Moreover,NAC could decrease manumycin-induced ROS generation and inhibit the effects of manumycin on the PI3K/Akt pathway and protect U937 cells from apoptosis induced by manumycin.Conclusion Manumycin induces apoptosis in U937 human leukemia cells through the regulation of the PI3K/Akt pathway and ROS production plays a critical role in the progress.
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OBJECTIVE:To optimize the extraction technology of Compound radix fici hirta granule. METHODS:Using the transfer rate of psoralen and amygdalin in extraction liquid of Compound radix fici hirta granule and extraction rate as indexes, U12(6×4×3)uniform design was used for the test,the effects of amount of adding water,extraction time,extraction times on the extraction technology were investigated,and optimized technology was verified by three pilot scale tests. RESULTS:The optimal technology was as follow as 10-fold water,extracting for 3 times,60 min each time. Under the conditions,transfer rate of pso-ralen and amygdalin and extraction rate were 82.51%(RSD=1.45%,n=3),93.69%(RSD=0.85%,n=3),18.89%(RSD=0.74%,n=3),respectively. The validated results were in the 95% confidence interval of predictive value. CONCLUSIONS:The optimized extraction technology is stable and feasible,and provides the scientific basis for the follow-up development of the prepa-ration.
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Objective To explore the effects and the mechanism of manumycin on KAT-18 cells Methods Human ATC (anaplastic thyroid cancer) KAT-18 cells were used. The cytotoxicity was analyzed by SRB assay. Apoptosis and cellular nitric oxide were detected by flow cytometry using annexin v and NO sensor dye. Superoxide anion was measured with a fluorescent plate reader by DHE.GSH was assayed by fluorescent Monochlorobimane. The SOD activities were assayed by colorimetric methods. The protein expression of Mn-SOD was determined by western blot. Results Manumycin decreased the viability of KAT-18 cells in a dose-dependent manner. Manumycin induced apoptosis significantly and NO generation simultaneously. Manumycin also induced superoxide anions generation. Manumycin reduced intracellular GSH in a time-course manner. However , manumycin did not decrease the SOD activity after 6 h treatment and Mn-SOD expression. A delayed induction of SOD activity was observed after 24 h manumycin treatment. N-acetyl-L-cysteine blocked NO and superoxide anions generation and apoptosis induction by Manumycin. Furthermore , NAC protected KAT-18 cells from the cytotoxicity of manumycin. Conclusion Manumycin induces apoptosis and has cytotoxic effects on KAT-18 cells. Cellular NO and superoxide anions generation are required for Manumycin-induced apoptosis in KAT-18 cells.
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Objective:To investigate the expression of CD56 antigen in leukemia cells of the patients with acute myeloid leukemia (AML)and its relationship with the prognosis of AML, and to clarify the role of CD5 6 antigen expression in predicting the prognosis of the AML patients.Methods:171 AML (non-M3)patients aged from 14 to 60 years old,who received a IA Regimen as the first time inducing chemotherapy were chosen.Flow cytometric analysis was used to evaluate the CD56 expression in leukemia cells.COX proportional regression analysis was used to select the prognostic factors,and bivariable analysis was used to study the relationship between the positive rate of CD56 and overall survival (OS).The CD56+ group (n=52),including CD56≥50% expression group (n=39) and CD560.05),while the 2-year survival rate in CD56≥50% group was lower than that in CD560.05).The relapse rate and first year relapse rate of patients in CD56+ group (64.3% and 37.5%)were significantly higher than those in CD56- group (34.3% and 17.9% )(P0.05).The DFS in CD56+ group was shorter than that in CD56- group (P<0.05).The same DFS result was also found between CD56≥50%group and CD56<50% group (P<0.05).Conclusion:The expression of CD56 antigen in leukemia cells predicts a bad prognosis in the AML patients,and the higher expression of CD56 indicates the worse prognosis.