Mesaconitine was incubated with rat liver microsomes in vitro. The metabolites of mesaconitine in rat liver microsomes were identified by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method with high resolution power. A typical reaction mixture of 100 mol L-1 Tris-HCI buffer (pH 7.4) containing 0.5 gL-1 microsomal protein and 50 micro molL-1 mesaconitine was prepared. The above reaction mixture was divided into six groups, and the volume of each group was 200 micro L. The incubation mixture was pre-incubated at 37 degrees C for 2 min and the reactions were initiated by adding NADPH generating system. After 90 min incubation at 37 degrees C, 200 micro L of acetonitrile was added to each group to stop the reaction. The metabolites of mesaconitine were investigated by UPLC-MS/MS method. Mesaconitine and 6 metabolites M1-M6 were found in the incubation system. The structures were characterized according to the data from MS/MS spectra and literatures. The metabolic reactions of mesaconitine in rat liver microsomes included the demethylation, deacetylation, dehydrogenation and hydroxylation. The major metabolic pathways of mesaconitine in rat liver microsomes were determined by UPLC-MS/MS on multiple reaction monitoring (MRM) mode combined with specific inhibitors of cytochrome P450 (CYP) isoforms, including alpha-naphthoflavone (CYP1A2), quinine (CYP2D), diethyldithiocarbamate (CYP2E1), ketoconazole (CYP3A) and sulfaphenazole (CYP2C), separately. Mesaconitine was mainly metabolized by CYP3A. CYP2C and CYP2D were also more important CYP isoforms for the metabolism reactions of mesaconitine, but CYP1A2 and CYP2E1 haven't any contribution to MA metabolism in rat liver microsomes.
Aconitine ; analogs & derivatives ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 CYP3A Inhibitors ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; metabolism ; Enzyme Inhibitors ; pharmacology ; Ketoconazole ; pharmacology ; Male ; Metabolic Networks and Pathways ; Microsomes, Liver ; enzymology ; metabolism ; Quinine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sulfaphenazole ; pharmacology ; Tandem Mass Spectrometry

Objective To investigate the feasibility and value of CT perfusion imaging in combination of CTA 320-slice CT with dual-low-dose in detection of acute cerebral ischemia.Methods Forty patients with acute stroke underwent perfusion imaging and CTA using a single scan with 320-slice CT with dual-low-dose schedule.CTA,4D-CTA,4D-perfusion imaging and fusion images were generated at a post-processing workstation.All patients also underwent a 3.0T magnetic resonance imaging with DWI for a comparison.Quality of the images,degree of vascular stenosis,and location of ischemic lesions were evaluated.Results The vascu-lar stenosis or occlusion was found in 33 patients,8 of whom were confirmed by the DSA.The ratio of better images of CTA was 82.5%.CTP showed 297 lesions with abnormal perfusion.202 lesions were confirmed as infarct by DWI,while 95 ones were nor-mal on DWI but with increased DLY and TTP and slightly decreased CBF and CBV in 49,increased DLY and TTP and normal CBF and CBV in 21,and increased DLY and TTP and slightly increased CBF and CBV in 25.Conclusion CT perfusion in combination with CTA by 320-slice CT with dual-low-dose is feasible and useful to observe accurately the vascular structures and the ischemic status of the whole brain at early stage.

Using a UPLC-MS/MS (MRM) and cocktail probe substrates method, the metabolic fingerprint of the compatibility of Radix Aconite (RA) and Radix Paeoniae Alba (RPA) and its effect on CYP450 enzymes were investigated. These main CYP isoforms include CYP 1A2, CYP 2C, CYP 2E1, CYP 2D and CYP 3A. Compared with the inhibition effect of RA decoctions on CYP450 isoforms, their co-decoctions of RA and RPA with different proportions can decrease RA' inhibition on CYP3A, CYP2D, CYP2C and CYP1A2, but can not reduce RA' effect on CYP2E1. The metabolic fingerprints of RA decoction and co-decoctions with different proportions of RPA in CYP450 of rat liver were analyzed by UPLC-MS. Compared with the metabolic fingerprints of RA decoction, the intensity of diester-diterpenoid aconitum alkaloids decreased significantly, while the intensity of monoester-diterpenoid alkaloids significantly increased in the metabolic fingerprints of co-decoctions of RA and RPA. The results suggest that RA coadministration with RPA increased the degradation of toxic alkaloid and show the effect of toxicity reducing and efficacy enhancing.
Aconitum ; chemistry ; Alkaloids ; chemistry ; Animals ; Chromatography, High Pressure Liquid ; Cytochrome P-450 Enzyme Inhibitors ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Liver ; drug effects ; enzymology ; Metabolome ; Paeonia ; chemistry ; Rats ; Tandem Mass Spectrometry

Partial nature of "promoting blood circulation and dieresis" of Salvia Miltiorrhizain was initially demonstrated by investigating the regulation effect of AQP2 expression in kidney of trauma blood stasis model rats with the Salvia Miltiorrhizain so as to provide guidance for its clinical deployment of administration. Random allocation was taken to averagely divide 30 SD rats into two groups: 10 rats in normal group and 20 rats in blood stasis syndrome group. Trauma blood stasis rat model was established by quantitatively beating. Then the rat model group was divided into model group and salvia group. After 7 days of treatment, the rat kidney AQP2 expression was detected, the content of urine AQP2 was compared and the damaged local muscle and kidney pathological changes were observed by immunohistochemical method and western blot method. Compared with that of the normal group, rats in model group had inflammatory cells infiltration, blood stasis and edema of the injured local muscles and up-regulated AQP2 expression, decreasing urinary output, and kidney tissues blood stasis and edema (P < 0.05). On the other hand, compared with that of the model group, those parameters of rats in salvia group were all decreasing except urine output (P < 0.05). Such result indicated that Salvia Miltiorrhiza can reduce trauma blood stasis rat content of urine AQP2 and down-regulated AQP2 expression in kidney tissue, so as to reduce the reabsorption of water by renal tubular and increase urine output. The promoting blood circulation effect of Salvia Miltiorrhizain can alleviate the degree of the damaged tissue edema and encourage urine drainage. This therapy is closely related to the effect of regulating AQP2 in kidney by salvia, so the purpose of this study by verifying "promoting blood circulation and diuresis" as the mechanism for the regulation effect of the salvia on AQP2 expression.
Animals ; Aquaporin 2 ; genetics ; metabolism ; Blood Circulation ; drug effects ; Diuresis ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Kidney ; blood supply ; drug effects ; metabolism ; physiopathology ; Kidney Diseases ; drug therapy ; genetics ; metabolism ; physiopathology ; Male ; Rats ; Salvia miltiorrhiza ; chemistry

High-speed counter-current chromatography (HSCCC) was used to high performance separate and prepare lignans from Schisandrae chinensis fructus. The solvent system is composed of n-hexane-ethyl acetate-methanol-water (9 : 1 : 5 : 5) and n-hexane-ethyl acetate-methanol-water (9 : 1 : 9 : 5), speed is at 900 r.min-1, and flow rate is at 2.0 mL.min-1. Five fractions from Schisandrae chinensis fructus extract were separated and prepared with one HSCCC process. They were identified as schisandrin, gomisin J, schisandrol B, schisantherin A and deoxyschizandrin by electrospray ionization-multiple tandem mass spectrometry (ESI-MSn), respectively. Their contents were obtained in 98.74%, 94.32%, 99.53%, 94.23% and 98.68% by ultra high performance liquid chromatography (UPLC), separately. The rapid and simple method can be applied for the preparation of lignans from Schisandrae chinensis fructus.
Countercurrent Distribution ; Cyclooctanes ; chemistry ; isolation & purification ; Dioxoles ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Fruit ; chemistry ; Lignans ; chemistry ; isolation & purification ; Molecular Structure ; Plants, Medicinal ; chemistry ; Polycyclic Compounds ; chemistry ; isolation & purification ; Schisandra ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

OBJECTIVE:To establish a method for simultaneous determination of the contents of Cu,Pb,As,Cr,Cd,Ba, Mn,Sb and Hg in Ganmaoling granule. METHODS:ICP-MS was conducted. The sample was handled by nitric acid microwave di-gestion system,the mass concnetration of 9 metal elements were determined by ICP-MS with Ge,In,Bi as internal standard and standard substance of test element standards. RF power was 1 100 W,sampling depth was 8 mm,carrier gas was argon(Ar)with high purity and flow rate was 1.0 L/min. RESULTS:All 9 elements had good linear range between mass concentration and ion peaks (r=0.999 1-0.999 9),detection limits were in the range of 0.3-6.0μg/L;RSDs of precision,stability and reproducibility tests were no more than 6.0%;average recoveries were in the range of 80.0%-113.5%,and RSDs were in 1.0%-4.5%. CONCLUSIONS:The method is simple,rapid and accurate,and can be used for the determination of 9 metal elements in Ganmaoling granule.

OBJECTIVE:To systematically review the efficacy and safety of pirfenidone in the treatment of idiopathic pulmo-nary fibrosis (IPF),and provide evidence-based reference for clinical treatment. METHODS:Retrieved from Cochrane Library, PubMed,EMBase,CJFD,CBM,VIP Database and Wanfang Database,randomized controlled trials (RCT) about the efficacy and safety of pirfenidone (test group) versus placebo (control group) in the treatment of IPF were collected,and Meta-analysis was performed by using Rev Man 5.1.7 software after data extracting and quality evaluating by modified Jadad. RESULTS:Totally 4 RCTs were enrolled,involving 1 153 patients. Results of Meta-analysis showed the decrease value of lung capacity [WMD=0.39,95%CI(0.16,0.61), P<0.001] and decrease value of the percentage of forced vital capacity to expected value [RR=0.68, 95%CI(0.53,0.87),P=0.002] in test group were lower than control group,there was significant difference between 2 groups;there was no significant difference in the lowest oxygen saturation [WMD=0.53,95%CI(-0.78,1.84),P=0.43] between 2 groups,however,the results of subgroup analysis showed the remission degree of 1 200 mg/d PFD for the decrease of lowest oxy-gen saturation was superior to placebo group,there was significant difference between 2 groups [WMD=1.72,95%CI(1.33,2.10), P<0.001],but therewas no significant difference between the remission degree of 1800 mg/d and placebo;and there was no signifi-cant difference in the incidence of adverse reactions between 2 groups [RR=1.70,95%CI(0.46,6.31),P=0.43],but the incidence of photosensitivity veactions in test group was significantly higher than that of control group,there was significant difference [RR=9.35,95%CI(4.23,20.67),P<0.001]. CONCLUSIONS:The efficacy of pirfenidone in the treatment of IPF is good,but the inci-dence of photosensitivity reactions should be noticed.

Objective A method for quantitative determination of 16 kinds of organochlorine pesticide (OCPs) in aquatic prod-ucts by gas chromatography (GC)was established .Methods The sample was extracted by acetonitrile ,purified by Carb/NH2 ,and then determined by GC .Results The linear relations of 16 kinds of OCPs were good at 0 .05 - 1 .00 mg/L(r> 0 .998) .The limit of detection of OCPs was in the range of 0 .04 - 0 .31 μg /kg(S/N = 3) .When the standard levels were 50 ,100 ,200 μg /kg ,the recovery rates were 72 .6% - 115 .2% ,the relative standard deviations were 0 .6% - 7 .5% .Conclusion The method established in this stud-y is applied to the determination of 16 kinds of OCPs in real samples with satisfactory results .

Objective: To establish a new method for determining residual organic solvent DMF in imiquimod. Methods: The samples were injected into an Agilent HP-PLOT/Q capillary column(30 m × 0. 530 mm,40. 0 μm) by a headspace sampler and ana-lyzed with an FID detector, the carrier gas was nitrogen, the inlet temperature was 250℃, and the detector temperature was 270℃. The column temperature was programmed raised. Results: The resolution among the peaks of DMF and the other residual solvents could meet the requirements. There was a good linearity within the experimental concentration range. The average recovery was 94. 6%(RSD=4. 0%, n=9). The limit of quantification and the limit of detection was 4. 809μg·ml-1 and 0. 963μg·ml-1, respectively. Conclusion:The method is convenient, accurate and sensitive, which can be used in the determination of residual solvent DMF in imi-quimod.

Objective:To compare the analgesic effect between living rhino horn and rhino horn in mice and rats,and to explore the possibility of living rhino horn used as a substitute of rhino horn. Methods:The analgesic effect was compared using the body tor-sion method and the formaldehyde method in mice,and the hot plate method and the thermal sting imager method in rats. Results:Compared with the control group,the living rhino horn at the dose of 0. 35,0. 7 and 1. 4 g·kg - 1 could significantly prolong the incu-bation period of body torsion induced by acetic acid(P < 0. 05 or P < 0. 01),and significantly reduce the number of body torsion(P <0. 05 or P < 0. 01). The three dose groups(0. 35,0. 7,1. 4 g·kg - 1 )of rhino horn could significantly reduce the number of body tor-sion(P < 0. 05 or P < 0. 01). After the second dose and compared with the control group,the pain threshold of high dose group(1. 4 g·kg - 1 )of living rhino horn,high and middle dose groups(0. 7,1. 4 g·kg - 1 )of rhino horn was significantly prolonged(P < 0. 05 or P < 0. 01). Compared with the control group,three dose groups(0. 175,0. 35,0. 7 g·kg - 1 )of living rhino horn and rhino horn could significantly reduce the analgesic effect in mice induced by formaldehyde in the second phase(P < 0. 01). Compared with the control group,the changes of pain threshold before and after the administration in three dose groups(110,220,440 mg·kg - 1 )of liv-ing rhino horn and high dose group(440 mg·kg - 1 )of rhino horn was significantly increased(P < 0. 05 or P < 0. 01). Conclusion:Living rhino horn can be used as a substitute of rhino horn with promising analgesia effect.