Objective To study the distribution and correlation of 20 kinds of inorganic elements (Na,Mg,A1,V,Cr,Co,Fe,Ni,Cu,Zn,Mn,As,Se,Cd,Cs,Sr,Ag,Hg,Pb,and U) in Jiawei Xiaoyao Formula (JXF).Methods The levels of 20 inorganic elements in 10 kinds of herbal substances were determined by ICP-MS with microwave digestion for sample preparation,and the results were analyzed with SPSS22.0.Results There were some correlations among the inorganic elements in JXF;Four main factors were selected by principal component analysis (PCA).The PCA results showed that Na,Mg,A1,V,Ni,Cu,Se,Sr,Cs,Mn,Hg,and Fe might be the characteristic elements in JXF;There were Mg,A1,Mn,and Fe elements in 10 herbal drugs,the contents of inorganic elements in herbal drug were different because the medicinal parts were different,and the decreasing order was roots and rhizomes,skin type,all grass,fruit seeds,and fungi.Conclusion Contents of inorganic elements can not only characterize the stability of extraction process,and can provide experimental evidence for the study of traditional Chinese medicine compound compatibility law.

Objective The visual inspection method were not appropriate to perform a hemolysis evaluation for colored injection like doxorubicin hydrochloride,this article adopted three methods to evaluate the hemolysis test of doxorubicin hydrochloride in vitro and provide reference for clinical drug safety.Methods Using rabbit erythrocytes as experimental object,the durg concentration 4.0 and 2.0 mg/mL was chosen which range of clinical concentration and preclinical safety evaluation concentration,to evaluate the hemolysis test of doxorubicin hydrochloride injection with blood analyzer test,direct colorimetric assay,and indirect colorimetric assay.Results The evaluation results of three different methods were very consistent.The tube's hemolysis rate of 4.0 mg/mL dose was far greater than 5％,which means serious hemolysis;Only 0.1 mL tube of 2.0 mg/mL dose (according to the drug concentration equal to 0.5 mL tube of 0.4 mg/mL drug concentration) without hemolysis occurring,the other tubes' hemolysis rates were far greater than 5％,which means serious hemolysis.Conclusion The hemolysis phenomenon may occur when 2.0 mg/mL dose of doxorubicin hydrochloride solution for iv injection is used in clinic and dilution (final concentration not more than 0.4 mg/mL) is recommended.

Objective To observe the protective and regeneration-promoting effects of Ruyi Zhenbao Pill (RZP) on nerve injury in zebrafish.Methods The zebrafish model of central nervous injury was induced by mycophenolate mofetil,the model of peripheral motor nerve and axonal injury was induced by ethanol,and the model of myelin damage was induced by ethidium bromide.The variations of central nerve,axon and myelin sheath fluorescence intensity,and peripheral motor nerve length in zebrafish,which exposed to the different concentration (10.0,33.3,and 100.0 μ.g/mL) of RZP,were observed with fluorescence microscope.The effective protection rates of RZP on zebrafish central nerve and axone,and the regeneration-promoting effect on peripheral motor nerve and myelin sheath were analyzed and calculated with the image processing software NIDS-Element's.Results In 10.0,33.3,and 100.0 μg/mL RZP groups,the zebrafish central nervous injury protective rates were 2％,24％,and 50％ (P ＜ 0.001),respectively,the peripheral nerve regeneration promoting rates were 44％ (P ＜ 0.05),49％ (P ＜ 0.01),and 93 ％ (P ＜ 0.001),the axonal injury recovery rates were 3％,29％ and 48％ (P ＜ 0.05),and the myelin sheath regeneration promoting rates were 36％ (P ＜ 0.01),37％ (P ＜ 0.001),and 41％ (P ＜ 0.001),compared with model group.Conclusions RZP could not only protect the central nervous and axonal injury,but also promote the regeneration of peripheral nerve and myelin sheath in zebrafish.

Objective To observe Rongxin Pills in the treatment of viral myocarditis in children (deficiency of both qi and yin and heart meridian stasis syndrome) and the effectiveness and clinical application of safety.Methods Viral myocarditis patients (280 cases,deficiency of both qi and yin and heart meridian stasis syndrome),according to 3:1 ratio as the test group (n =21 0) and control group (n =70).The test group took orally Rongxin pills each time 4.5～9 g,3 times daily;the control group oral coenzyme Q10 capsule each time 10 ～ 20 mg,twice daily.The course of treatment was 28 d.The experiment was carried out with the random and double blind method.The symptoms of myocarditis,integrated and electrocardiogram,echocardiography,myocardial enzymes,as well as the efficacy of traditional Chinese medicine and improvement of the effect of the disease were observed.Results The results of FAS (PPS) analysis showed that 28 d after treatment,the symptom score and mean of experimental group and control group were 5.975 (6.000) and 4.721 (4.788).The syndromes of the total effective rates were 91.62％ (90.59％) and 70.59％ (71.21％),curative effect the total effective rates were 90.14％ (92.08％) and 72.06％ (72.73％).The total effective rate of experimental group was higher than that of the control group,the difference was statistically significant.In this experiment,three cases of clinical adverse events were reported,which were not related to the experimental drug.It also not belongs to adverse drug reactions.Conclusion Rongxin Pill in the treatment of viral myocarditis in children (deficiency of both qi and yin and heart meridian stasis syndrome) is more effective than coenzyme Q 10 capsule,and there was no indication of higher risk of clinical application.

Objective To establish a method for determination of the twelve residual organic solvents,including methanol,ethanol,acetone,isopropanol,tert-Butyl methyl ether,dichloromethane,aceticether,tetrahydrofuran,triethylamine,trimethylorthofor-Mate,morpholine,N,N-Dimethylformamide in Apixaban bulks drug.Methods Gas head-space chromatography was applied to this study.The column was DB-624 silica capillary column (30.0 m × 0.53 mm × 3.00 μm) and the carrier gas was high purity nitrogen;The vial temperature was 100 ℃,and the vial time was 20 min.The Column temperature was kept at 40 ℃ for 6 min,then the temperature was raised to 220 ℃ at the rate of 20 ℃/min and subsequently sustained for 10 min.FID detector temperature and injection temperature were both 250 ℃.The N2 flow rate was 2.8 mL/min.Split ratio was 5∶1.Results Twelve kinds of solvents were completely separated and determined with a good linearity (r =0.9994-0.9999).The RSD values of precision experiments and the average recovery was in line with the requirements.Conclusion Theanalytical method is simple,accurate and sensitive,which could be used for determination of residual organic solvents in Apixaban bulks drug.

Objective To study the inhibitory effect of Curcumin combined with Glycyrrhetinic acid on proliferation of HepG-2 cell,and probe the reasonability and scientificity of curcuma combined with glycyrrhiza.Methods The CCK-8 method was used to test the proliferation inhibition rate of HepG-2 cells after treated with Curcumin (10.00,5.00,2.50,and 1.25 μg/mL),Glycyrrhetinic acid (20.0,10.0,5.0,and 2.5 μg/mL) and Curcumin combined with Glycyrrhetinic acid in corresponding concentration for different time (8,16,or 24 h).Flow cytometry was used to detect the cell apoptosis and the cell cycle of HepG-2 cells after treated with Curcumin (5 μg/mL),Glycyrrhetinic acid (10 μg/mL) and Curcumin combined with Glycyrrhetinic acid in corresponding concentration for 24 h.Results Curcumin,Glycyrrhetinic acid and drug combination obviously inhibited the proliferation of HepG-2 cells in a time-and concentration-dependent manner,the effects of combination group was more stronger and showed additive effect or synergistic effect.The apoptosis rate of HepG-2 cells was significantly increased after treated with three groups of drug,combination group showed additive effect;Curcumin,Glycyrrhetinic acid and the drug combination showed significant G2 arrest.Conclusion Curcumin combined with Glycyrrhetinic acid has positive effect on inhabiting the proliferation and promoting apoptosis of HepG-2 cell.Curcuma combined with glycyrrhiza possess reasonability and scientificity.

Objective To study the inhibitory effect on proliferation of Hela cells of podophyllotoxin,4'-demethylepi-podophyllotoxin and drug combination of different proportion of nano-sillca (SiO2) and 4'-demethylepi-podophyllotoxin in vitro,and discuss the mechanism.Methods Used ethyl silicate hydrolysis method to prepare 25 nm SiO2 sample,next carried 4'-demethylepi-podophyllotoxin after the surface modification,and measure cell campatibility by MTT method and Hoechst 33342.The inhibitory effect of podophyllotoxin,4'-demethylepi-podophyllotoxin and drug combination on proliferation of Hela cells was measured by MTT assay.Hoechst 33342 staining method was used to detect cell apoptosis.The effect ofdmg combination treatment on cell morphology was observed by inverted microscope.Western blotting technique was used to detected effect of 4'-demethylepi-podophyllotoxin and drug combination on expression of apoptosis related protein.Results Inhibitory effect onproliferation of Hela cells of 4'-demethylepi-podophyllotoxin is superior to podophyllotoxin,inhibitory effect of drug combination is superior to the single 4'-demethylepi-podophyllotoxin,the inhibition of drug combination with 0.125 μg/mL nano SiO2 and 6.25 μg/mL 4'-demethylepi-podophyllotoxin is the most obvious.MTT and Hoechst 33342 experimental results showed that the 25 nm SiO2 have good cell compatibility.Podophyllotoxin,4'-demethylepi-podophyllotoxin and drug combination can induce apoptosis.Western blotting results showed that 4'-demethylepi-podophyllotoxin and drug combination can up-regulate the ratio of Bax/Bcl-2 and the expression level of Caspase-3、P53 and P38.Conclusion In vitro experimental performance of drug combination is superior to single 4'-demethylepi-podophyllotoxin,it is may by effecting the expression of Bcl-2,Bax,Caspase-3,P53,and P38 and others apoptosis related protein to induce Hela cell apoptosis.

Objective The three-dimensional quantitative structure activity relationship (3D-QSAR) method was applied to study thiazole derivatives as potent inhibitors ofdihydroorotate dehydrogenase,which provided useful guidance for more discovery of potent inhibitors of dihydroorotate dehydrogenase.Methods Molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) were applied to systematicly investigate 3D-QSAR of 38 hiazole derivatives as potent inhibitors of dihydroorotate dehydrogenase.Established models of CoMFA and CoMSIA and the predictive ability of models were validated.Three dimensional map was applied to analyzing the relationship between structure and activity of thiazole derivatives.Results The coefficients of cross validation q2 and non-cross validation r2 for CoMFA model were 0.796 and 0.978,and for CoMSIA model were 0.721 and 0.976 respectively.The prediction of activity of compound was close to the actual value of the two models.Effect of compound structure on its activity could be analyzed comprehensively and intuitively by three dimensional map.Conclusion The model reveals the relationship between the structure characteristics and the inhibitory activity,and has good predictive capability and stability to lay a good foundation for further development and research.

Objective Using middle cerebral artery occlusion (MCAO) model to observe protective effect of effective components of bezoar on brain damage.To discuss the anti-cerebral ischemia mechanism and compare the efficacy of effective components of bezoar from the endoplasmic reticulum stress intervention angle.Methods Rats were stratified randomly divided into sham group,model group,Qingkailing group (positive drug,3 mL/kg),taurine group,ursodeoxycholic acid (UDCA,78 mg/kg) group,taurine-conjugated ursodeoxycholic acid (TUDCA,100 mg/kg) group.Through establish MCAO model in rats,observed the scores of the neurologic impairment,measured infarct volume by TTC.Immunohistochemistry and Western blotting were Used to detect the content of P-PERK,P-EIF2α,and ATF4.Results Compared with sham group,neurologic impairment scores of model group significantly reduced (P ＜ 0.01).Compared with model group,Qingkailing group,UDCA group,and TUDCA group significantly improved neurological function in rats (P ＜ 0.05,0.01).Compared model group,all the treatment groups could significantly reduce the volume of cerebral infarction (P ＜ 0.01).Compared with sham group,expression of P-PERK,P-EIF2α,and ATF4 was significantly increased (P ＜ 0.01).Compared with model group,all the treatment groups reduced the expression of P-PERK,P-EIF2α,and ATF4 in varying degrees,effect of Qingkailing and TUDCA were more obvious.Conclusion The effective components ofbezoar alleviate cerebral ischemia reperfusion injury in rats by inhibiting endoplasmic reticulum stress,the effect of TUDCA is better than that of taurine and UDCA.

Objective To study the inhibitory effects of TPP,a desmosdumotin-C B ring para-fluoro modified derivative,on human breast cancer MCF-7 cell proliferation,and investigate the possible mechanisms.Methods MTT assay was used to measure the proliferation suppression of MCF-7 cells after treated with 1.0,2.5,5.0,10.0,and 20.0 μg/mL TPP for 48 h,and then the cell apoptosis rate and expression rate of NF-κB P65 positive cells were tested by flow cytometry after 20.0 μg/mL TPP treatment for 0,24,and 48 h.Results MTT assay showed that,after treatment for 48 h,1.0,2.5,5.0,10.0,and 20.0 μg/mL TPP all exhibited the inhibitory effects and showed a dose-dependent relationship.Flow cytometry results showed that 20.0 μg/mL TPP induced cell apoptosis after treatment for 24 and 48 h.TPP (20.0 μg/mL) significantly reduced the rate ofNF-κB P65-positive cells in MCF-7 cells after treatment for 48 h.Conclusion TPP could inhibit the proliferation of MCF-7 cells,which may be induced by cell apoptosis.Down-regulation of NF-κB is possible to be related with apoptosis.