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

In order to optimize extraction process conditions of tannins from Geranium orientali-tibeticum by supercritical CO2, the content of tannins was determined by phosphomolybdium tungsten acid-casein reaction, with extraction pressure, extraction temper- ature and extraction time as factors, the content of tannins from extract of G. orientali-tibeticum as index, technology conditions were optimized by orthogonal test. Optimum technology conditions were as follows: extraction pressure was 25 MPa, extraction temperature was 50 °C, extracted 1.5 h. The content of tannins in extract was 12.91 mg x g(-1), extract rate was 3.67%. The method established could be used for assay the contents of tannin in G. orientali-tibeticum. The circulated extraction was an effective extraction process that was stable and feasible, and that provides a way of the extraction process conditions of tannin from G. orientali-tibeticum.
Carbon Dioxide ; chemistry ; Chemical Fractionation ; methods ; Drugs, Chinese Herbal ; chemistry ; Geranium ; chemistry ; Tannins ; isolation & purification

Objective: To evaluate the efficacy and safety of acupuncture for the treatment of knee osteoarthritis and to provide evidence for its use in clinical practice. Methods: Eight databases were extensively searched up to March 2018. Randomized controlled trials (RCTs) comparing the efficacy of acupuncture with sham acupuncture or no acupuncture for knee osteoarthritis were included. Study selection, data extraction and quality assessment were independently conducted by two reviewers. The Cochrane Collaboration's tool was used for assessing the risk of bias. Results: A total of 18 RCTs were included, involving a total of 3522 participants. The results showed that acupuncture was superior to sham acupuncture in relieving pain (SMD=-0.34, 95％CI:-0.57 to -0.11, I2=85％, P=0.003) and improving physical function (SMD=-0.34, 95％CI: -0.57 to -0.11, I2=85％, P=0.003). In comparison to the no-acupuncture group, the acupuncture group also showed significant advantages in relieving pain (SMD=-0.79, 95％CI: -1.15 to -0.43, I2=87％, P<0.0001) and improving physical function (SMD=-0.75, 95％CI:-1.19 to -0.31, I2=91％, P=0.0008). Sensitivity analyses suggested that the results were robust, and Egger's test found no potential publication bias. Conclusion: In the treatment of knee osteoarthritis, the acupuncture group had significant advantages over sham acupuncture or no-acupuncture groups in relieving pain and improving physical function.

Objective To investigate the level of serum free fatty acid (FFA )after improving the life style in patients with coronary heart disease complicated metabolic syndrome and the effect of therapeutic life style on traditional risk factors of coronary artery disease.Methods A total of 395 patients with coronary heart disease complicated metabolic syndrome were recruited.Pa-tients were divided into intervention group (group A,conventional drug therapy+ intensive life style intervention,n=97)and non-intervention group (group B,conventional drug therapy,n=38)according to the scores of life style.Serum free fatty acid (FFA) was determined by ELASA.The scores of life style was obtained bylife style questionnaire.Results (1)The serum FFA of pa-tients with coronary heart disease complicated metabolic syndrome were positively related to waist circumference and waist-high-ra-tio.(2)Waist circumference,BMI and FFA of group A were significantly lower than those in group B after therapeutic life style in-tervention(P <0.05).(3)Compared with the baseline,the constitution index and FFA in group A were significantly lower after 6-months therapeutic life style intervention(P <0.05).Conclusion Therapeutic life style can reduce the level of FFA and constitution index of the patients with coronary heart disease complicated metabolic syndrome.

Accidents, Traffic ; Forensic Medicine ; Hoarseness ; Humans

OBJECTIVETo compare the differences of active ingredients between tissue cultured cells and cultivated saffron pistils.

METHODThe experiment was carried out by the Fourier transform infrared spectroscopy (FTIR) spectra and high performance liquid chromatography (HPLC).

RESULTThe data indicated that the species and contents active ingredients in saffron pistils from different places were different. The species of active ingredients in tissue cultured cells are less than those in cultivated saffron pistils. However, the quantity of crocin A, which showed good anticancer effect, is 2-3 times more than that in cultivated saffron pistils.

CONCLUSIONThe active ingredients of the tissue cultured cells are similar to those of saffron pistils, but their contents are different. Therefore, the tissue cultured cells can only be the part-substitutes of cultivated saffron pistils.

Carotenoids ; analysis ; Cells, Cultured ; Chromatography, High Pressure Liquid ; Crocus ; chemistry ; cytology ; Ecosystem ; Flowers ; chemistry ; cytology ; India ; Plants, Medicinal ; chemistry ; cytology ; Spain ; Species Specificity ; Spectroscopy, Fourier Transform Infrared

Objective To analyze the clinical features,therapeutic effect and prognosis in patients with bone pain induced by malignant bone me?tastasis as well as the rationality of analgesic application,so as to improve the level of diagnosis and treatment for metastatic bone pain. Methods Totally 123 patients with pain due to malignant bone metastasis received antitumor therapy and analgesic therapy based on standardized three?step guidelines. Their clinical characteristics were retrospectively analyzed. Results The total pain relief rate was 85.4%and the pain was significantly relieved(P<0.05). The DUI value of each narcotic agent was close to 1 and the application of narcotic agents tended to be rational. The Kaplan?Meier survival analysis showed that patients with moderate pain had longer survival time than those with severe pain(P=0.015). The survival rate of patients with significant pain relief after treatment was higher than those unrelieved(P=0.021). The survival rate of patients without visceral me?tastasis was higher than those with visceral metastasis(P=0.000). The COX multivariate analysis indicated that the pain intensity and visceral me?tastasis were independent risk factors influencing patient prognosis. Conclusion Standard treatment can improve symptoms in most patients with bone metastasis and prolong survival time. Opioids have satisfactory analgesic effect for moderate to severe pain and the adverse reactions can be tol?erated.

Objective To clarify the role and significance of Toll?like receptor 4(TLR4)in myocardial damage following paraquat(PQ)poisoning in mice. Methods Male wild type C57BL/6J mice(WT)and male TLR4 deficient mice(TLR4?ko)were divided into four groups in the study:(1)control group(WT mice,n=6);(2)TLR4?ko group(TLR4?ko mice,n=6);(3)WT+PQ group(WT mice,n=30);(4)TLR4?ko+PQ group (TLR4?ko mice,n=30). The mice in group 1 and group 2 were injected intraperitoneally with saline;mice in group 3 and group 4 were injected in?traperitoneally with 75 mg/kg of PQ. At 2 h,4 h,8 h,16 h and 24 h after PQ administration,6 mice of WT+PQ group and TLR4?ko+PQ group were euthanized and the heart tissue specimens were harvested. All the specimens were analysed by histology,while the expression of TLR4 mRNA was only detected in samples of WT+PQ mice. The specimens at 2 h,8 h and 24 h after PQ administration were used for cytokines detection for WT+PQ group and TLR4?ko+PQ group;in addition,Western blot analysis was performed for WT+PQ group. At 8 h after treatment,control mice and TLR4?ko mice were euthanized by the same method. Mice were anesthetized for cardiac geometry and functional assessment using a 2?D guide M?mode echocardiography at 8 h following injection of either PQ or saline. Results During myocardial damage due to PQ exposure in WT+PQ mice,obvious histopathological changes were observed,as well as a noticeable decrease of heart function and increased expressions of TNF?αand IL?1β. Compared with the WT mice,TNF?αand IL?1βprotein levels,changes in heart function and histopathological changes were significantly attenuated following PQ exposure in myocardial damage in TLR4?ko mice. Conclusion The TLR4gene is involved with in heart functional injury and histopathological changes in myocardial damage following PQ poisoning in mice,which may through the regulation of TNF?αand IL?1βexpression. Our findings indi?cate that TLR4 plays an important role in mediating myocardial injury due to PQ.

1.0?108 co-pies/L)and the lower viral load group(HBV DNA