Determination of Dendrobine and Its Metabolites M-250 and M-280 in Mice Plasma by UPLC-MS/QAMS / 中国药房

OBJECTIVE:To establish a method for the determination of dendrobine and its metabolites M-250 and M-280 in mice plasma for the first time. METHODS:Mice were given dendrobine 60 mg/kg by intragastric administration,1 h later plasma were collected and treated. Using pseudoephedrine hydrochloride as internal standard and dendrobine reference substance as control, the plasma concentrations of dendrobine and its metabolites M-250 and M-280 were determined by UPLC-MS combined with quantitative analysis of multi-components by single marker. The separation was performed on Hypersil Gold C18 column with 0.1％formic acid-acetonitrile(gradient elution)at the flow rate of 0.3 mL/min. The column temperature was set at 40℃,and sample size was 5 μL. Heatable electrospray ionization (HESI) source, scan/ESI + were applied and operated in positive ion mode with atomization temperature of 300℃,ion transmission tube temperature of 350℃,the sheath gas velocity of 35 arb,the auxiliary air velocity of 15 arb,the spray voltage of 3.5 kV,the collision voltage of 30,40,50 eV. The mass-to-charge ratio of detection range were 100-1500. RESULTS:The endogenous substances of mice plasma had no interference with the content determination of dendrobine and its metabolites M-250 and M-280. The linear range of dendrobine were 9.13-912.94 ng/mL(r=0.9996). The limit of quantitation was 3.04 ng/mL. RSDs of intra-day and inter-day were all less than 7.5％(n=5 or n=3). The accuracy were 96.8％-107.5％(n=5). Matrix effects were 97.1％-106.0％(RSD=1.8％-4.7％,n=5). RSDs of the content of sample at 15℃ for 24 h,at -70 ℃ after three times freeze-thaw,at -70 ℃ for 15 d were lower than 12.8％ (n=3). The content of dendrobine in plasma sample of mice was (41.3 ± 5.7) ng/mL (n=12). The contents of its metabolites M-250 and M-280 were (493.0 ± 73.1) and (41.4 ± 3.0) ng/mL (n=12) with Relative correction factor of 1.0. CONCLUSIONS: The method is sensitive and accurate,and can be used for content determination of dendrobine and its metabolites M-250 and M-280 in mice plasma.

Effects of rifampicin on hepatotoxicity and genes related to bile acid metabolism in mice / 中国药理学通报

Aim Toexamineliverdamagebyrifampi-cin and hepatic gene expression related to bile acid me-tabolisminmice.Methods Adultmalemicewere given rifampicin(180 mg·kg-1 ,po)daily for 30 days and(90 mg·kg-1 ,po)daily for 90 days,blood bio-chemistry,histopathology,and gene expression were examined.Results Rifampicinincreasedanimalliver index and serum enzyme activities. Histopathology showed steatosis and spotted feathery-like degenera-tion.Rifampicin increased the expression of CYP7A1 after 30 and 90 days of administration,along with in-creased FXR and SHP.Rifampicin reduced the expres-sion of BSEP after 30 days of high dose administration. Conclusion Repeatedadministrationofrifampicin may cause liver injury and intrahepatic cholestasis in mice,and these effects are associated with the altera-tion of gene expression related to bile acid metabolism.

Effect of nickel-titanium wire on the expression of sarcoplasmic reticulum Ca~(2+)-ATPase and ryanodine receptor in rat myocardial cells / 中国组织工程研究

BACKGROUND:Although the nickel-titanium occluder in the treatment of congenital heart disease has a better clinical effect, arrhythmia wil be more likely to develop in late stage. OBJECTIVE:To investigate the effect of nickel-titanium wire on expression of sarcoplasmic reticulum Ca~(2+)-ATPase and ryanodine receptor of rat myocardial cels. METHODS:Thirty Sprague-Dawley rats were obtained and randomly divided into two groups: experimental and control groups. The nickel-titanium wire was implanted to the apex of heart of rats in the experimental group. Rats in the control group received no special treatment. Rat mycardial cels were harvested at the 1th, 3rdand 6th months after operation. The gene and protein expressions of sarcoplasmic reticulum Ca~(2+)-ATPase and ryanodine receptor of rat myocardial cels were detected using RT-PCR and immunohistochemistry, respectively. The inflammatory reactions were detected using hematoxylin-eosin staining. RESULTS AND CONCLUSION:After the nickel-titanium wire was implanted into the rat myocardium, inflammatory reaction was induced by inflammatory cel infiltration in the experimental group, with hyperplasia of fibrous tissue. The inflammatory reaction gradualy disappeared as the implanted time extended. No inflammatory cel infiltration was visible in the control group. There was no significant difference in the gene and protein expressions of sarcoplasmic reticulum Ca~(2+)-ATPase and ryanodine receptor of rat myocardial cels at different time points after operation between these two groups. It showed that nickel-titanium wire had no influence on the expressions of sarcoplasmic reticulum Ca~(2+)-ATPase and ryanodine receptor of rat myocardial cels. These results suggest that nickel-titanium occluder-related arrhythmia may have little relationship with abnormal protein expression of sarcoplasmic reticulum Ca~(2+)-ATPase and ryanodine receptor.

Safety evaluation of realgar-and cinnabar-containing traditional Chinese medicine / 中国中药杂志

Realgar (90% of AS4S4) and cinnabar (96% of HgS) have been used in traditional Chinese medicines for thousands of years. Both arsenic and mercury are well-known for toxic effects and the safety of realgar-and cinnabar-containing traditional Chinese medicines is of concern. It is considered that any intentional use of known toxic metals in medicine is an unacceptable risk, while an opposing opinion presumes that realgar and cinnabar have clear pharmacological action with tolerable side effects. This review summarized the progress of toxicological study on realgar-and cinnbar-containing traditional Chinese medicines.

Realgar is active ingredient of Angong Niuhuang pill in protection against LPS-induced neuroinflammation / 中国中药杂志

<p><b>OBJECTIVE</b>To determine the effects of Angong Niuhuang pill (AGNHW) on lipopolysaccharide (LPS)-induced neuroinflammation and further to investigate the role of realgar and cinnabar on AGNHW-mediated neuroprotection.</p><p><b>METHOD</b>Primary rat midbrain neuron-glia cultures were used as an in vitro model to examine the effects of AGNHW on LPS-induced dopamine (DA) neuronal damage. Cultures were divided randomly into five groups: control, LPS, LPS plus AGNHW, LPS plus realgar and LPS plus cinnabar. Dopaminergic neurotoxicity was measured by [3H] DA uptake assay. The production of intracellular reactive oxygen species (ROS) was quantified via the DCFH-DA probe. Real-time RT-PCR was applied to detect the mRNA expression of pro-inflammatory factors. Then the protein levels of these factors were determined by ELISA and western blot assay.</p><p><b>RESULT</b>Compared with the control group, LPS apparently decreased DA uptake capacity (P < 0.05); induced the production of intracellular ROS (P < 0.05); enhanced the mRNA expression of TNF-alpha, iNOS, IL-13 and COX-2 (P < 0.05) and the release of TNF-alpha, IL-1beta and PGE2 in the supernatant of cultures (P < 0.05); and also increased the level of iNOS protein (P < 0.05). Compared with the LPS group, AGNHW and realgar significantly inhibited LPS-induced reduction of DA uptake (P < 0.05); attenuated the production of intracellular ROS (P < 0.05) and the mRNA expression of TNF-alpha, iNOS, IL-1beta and COX-2 (P < 0.05) and the release of TNF-alpha, IL-1beta and PGE2 (P < 0.05) and the level of iNOS protein (P < 0.05). However, there was no significant difference between LPS group and LPS plus cinnabar group.</p><p><b>CONCLUSION</b>AGNHW is effective in protecting against LPS-induced neuroinflammation, and realgar is one of active components for AGNHW to produce anti-inflammatory effects.</p>

Experience of introducing analysis of clinical cases into experiment teaching of pharmacology / 中华医学教育探索杂志

Introduction of clinical cases to experiment teaching of pharmacology is an effective method,which combines basic theory with clinical medicine.It can stimulate students' interest in pharmacology,enhance their go-aheadism in study and also improve teaching effects.Moreover,it plays an important role in improving the students qualities such as the reasoning ability,the verbal capacity and the ability to analve and resolve problems,etc..In this article,we explored the teaching method in experiment teaching of pharmacology,and evaluated the effect by questionnaire.

Effects of Breviscapine on the contraction of rabbit aorta strips induced by noradrenaline and high potassium / 中国药理学通报

Aim To observe the effects of Breviscapine (Bre) on the contractions induced by noradrenaline (NA) and high potassium in rabbit aorta strips and to investigate the relationships of these effects to the changes of intracellular free calcium( i). Methods The effects of Bre on the concentration-response curves for NA and high potassium, and on the transient contractions induced by NA and caffeine in Ca 2+-free medium and the sustained contraction by NA after replenishing Ca 2+ were surveyed using rabbit aorta strips; the changes of i increased by NA and high potassium in the presence of Bre were determined using fura-2/AM loaded cultured smooth muscle cells of rabbit aorta. Results Bre shifted the concentration-response curve for NA to right in a dose-dependent manner , but shifted that for high potassium to left; it inhibited the transient contraction induced by NA and caffeine in the Ca 2+-free medium and the sustained contraction induced by NA after replenishing Ca 2+; Bre inhibited the i increased by NA, but enhanced that increased by high potassium in the smooth muscle cells of rabbit aorta. Conclusion Bre inhibits the contraction induced by NA through its inhibition effects on Ca 2+-influx and Ca 2+-release ; it enhances the Ca 2+-influx induced by high potassium, but the mechanism by which Bre enhances the high potassium inducing Ca 2+ -influx is not known.

Study on the grafting skin organ of immunoregulation by warm moxibustion meridian / 中国免疫学杂志

Chinese traditional warm moxibustion meridian was used to enhance the regulationofimmune function.The survival period of grafting skin was determind. Using these indexes the anthor oberved whether the reaction of grafting skinwas decreased and the variety of immune reaction to the T Lymphocytes,B Ly-mphocytes and Macrophage in the spleen and serum of mice were also observed The primary results were as follows. 1. warm moxibustion meridian can prolong the survival period of grafted skin.Thegroup of control were 10.4?2.5days.Governor Vessl Meridian,23.2?2.5(GV).BladderMeirdian(B)16.5?3.5;Governor Vessel Meridian and Bladder Meridian,(GV+B):24.5?4.5.P