Bile acids(BAs)are synthesized by the liver from cholesterol through several complementary pathways and aberrant cholesterol metabolism plays pivotal roles in the pathogeneses of cholesterol gallbladder polyps(CGP)and cholesterol gallstones(CGS).To date,there is neither systematic study on BAs profile of CGP or CGS,nor the relationship between them.To explore the metabolomics profile of plasma BAs in healthy volunteers,CGP and CGS patients,an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was developed and validated for simultaneous determination of 42 free and conjugated BAs in human plasma.The developed method was sensitive and reproducible to be applied for the quantification of BAs in the investigation of plasma samples.The results show that,compared to healthy volunteers,CGP and CGS were both characterized by the significant decrease in plasma BAs pool size,furthermore CGP and CGS shared aberrant BAs metabolic characteristics.Cheno-deoxycholic acid,glycochenodeoxycholic acid,λ-muricholic acid,deoxycholic acid,and 7-ketolithocholic acid were shared potential markers of these two cholesterol gallbladder diseases.Subsequent analysis showed that clinical characteristics including cysteine,ornithine and body mass index might be closely related to metabolisms of certain BA modules.This work provides metabolomic information for the study of gallbladder diseases and analytical methodologies for clinical target analysis and efficacy evaluation related to BAs in medical institutions.

Pyrrolizidine alkaloids(PAs)are among the most hepatotoxic natural compounds that are widely distributed throughout the world.Most PAs are metabolically activated to trigger toxicity.Exposure to herbal medicine containing PAs and food supplements contaminated by PAs is considered to be one of the two main causes of hepatic sinusoidal obstruction syndrome(HSOS),which is a rare hepatic vascular disease with a high mortality rate.PAs-induced HSOS cases have been reported worldwide.However,there is no clinically effective therapy for PAs-induced HSOS,which is partially because the toxic mechanism is not fully understood.This review focuses on updating the information on the metabolism and the molecular mechanisms of PAs hepatotoxicity,including oxidative stress,apoptosis,and dysfunction of bile acid metabolism,and their interactions.

OBJECTIVE:To explore the role of clinical pharmacists participating in the individualized treatment for purulent meningitis. METHODS:Clinical pharmacists participated in the therapy for a patient with purulent meningitis complicated with Staphylococcus aureus infection. According to patient's condition,clinical pharmacists assisted physicians to formulate preliminary therapeutic plan. Reviewing relevant guidelines,domestic and foreign literatures,clinical pharmacists suggested to combine with dexamethasone so as to relieve inflammatory reaction. According to the results of drug sensitivity test,based on vancomycin plasma concentration monitoring and population pharmacokinetics model fitting,clinical pharmacists suggested to reduce the dose of vanco-mycin to 0.5 g,ivgtt,q12 h. The pharmaceutical care were conducted throughout the therapy,including efficacy evaluation of an-ti-infective therapy,ADR monitoring,renal function monitoring,etc. RESULTS:Physicians adopted some suggestions of clinical pharmacists. The disease condition of the patient was recovred,and no ADR related to vancomycin was found. On the 16th day, the patient was discharged from the hospital. CONCLUSIONS:Clinical pharmacists participate in treatment of purulent meningitis, assist physicians to optimize therapy plan based on relevant guideline,literature,etiological examination,blood concentration moni-toring and pharmacokinetics model fitting results. It not only guarantee therapeutic efficacy of anti-infective therapy,but also pre-vent and reduce the occurrence of ADR.

OBJECTIVE:To explore the role of clinical pharmacists in therapy for patient with interstitial lung disease (ILD) induced by erlotinib. METHODS:Clinical pharmacists participated in the therapy for ILD in a patient receiving erlotinib target treat-ment after thoracic vertebra and lumbar radiation,analyzed the cause of ILD and suggested to stop taking imipenem and cilastatin sodium,fluconazol and erlotinib according to lab indexes and patient’s symptom;took prednisone 30 mg,po,qd,for anti-inflam-mation instead of methylprednisolone;adjusted the dose of prednisone to 40 mg/d,and additionally took Carbocisteine oral solution 10 ml,tid,for improving respiratory symptom;panipenem betamipron 1 g,ivgtt,bid,instead of piperacillin sodium and sulbactam sodium. RESULTS:Physicians adopted the suggestions of clinical pharmacists,and the symptom of anhelation and double pneumo-nia recovered;discharged medication plan was erlotinib 150 mg,po,qd. CONCLUSIONS:The patient with radiation history easily suffers from ILD when using erlotinib,and should use erlotinib carefully in the clinic. Clinical pharmacists participated in drug ther-apy and promote safe and rational use of drugs in the clinic.

Aim To explore micro RNAs-integrated pathogenic signaling to control endothelial-mesenchy-mal transition ( EndMT ) in pulmonary hypertension ( PH) by a network bioinformatic approach. Methods Literature-mining method was used to find PH-relat-ed genes and EndMT/EMT-related miRNAs. Bioinfor-matic prediction approach ( DIANA3 , Miranda4 , PicT-ar5 , TargetScan6 , miRDB7 and microT-CDS8 ) was used for miRNA target prediction. Hypergeometric a-nalysis was used to predict miRNAs related to EndMT in PH. The analysis of interactions between PH-rele-vant genes( PH network) was performed with the use of Biological General Repository for Interaction Datasets ( BioGRID ) . These miRNAs were ranked with the highest probability of substantial overlap among their gene targets in the PH-network, the relationship be-tween their targets and the PH functional categories which include hypoxia, inflammation, and transforming growth factor/BMP signaling. Then, the part of results was validated by animal experiment. Lastly the miR-NA-Target network was built using Cytocape 3 . Results List of 230 genes was compiled that were directly im-plicated in the development of PH and 189 miRNAs were related to EndMT in PH. Among 189 miRNAs, only 22 microRNAs(miR-let-7 family, miR-124, miR-130 family, miR-135, miR-144, miR-149, miR-155, miR-16-1, miR-17, miR-181 family, miR-182, miR-200 family, miR-204, miR-205, miR-21, miR-224, miR-27, miR-29 family, miR-301a, miR-31, miR-361 and miR-375) were related to hypoxia, inflamma-tion, and transforming growth factor/BMP signaling. Among these miRNAs, the levels of let-7g, miR-21, miR-124 and miR-130 family were significantly changed in the pulmonary artery in hypoxia-induced PH rats. Conclusions Among numerous miRNAs,22 of which may be involved in hypoxia, inflammation, and transforming growth factor/BMP signaling and re-lated to EndMT in PH by network bioinformatic ap-proach, which provides a theoretical basis for further investigation of EndMT in PH.

OBJECTIVE: To explore the role of calpain in pulmonary vascular remodeling in hypoxia-induced pulmonary hypertension and the underlying mechanisms.
 METHODS: Sprague-Dawley rats were randomly divided into the hypoxia group and the normoxia control group. Right ventricular systolic pressure (RVSP) and mean pulmonary artery pressure (mPAP) were monitored by a method with right external jugular vein cannula. Right ventricular hypertrophy index was presented as the ratio of right ventricular weight to left ventricular weight (left ventricle plus septum weight). Levels of calpain-1, -2 and -4 mRNA in pulmonary artery were determined by real-time PCR. Levels of calpain-1, -2 and -4 protein were determined by Western blot. Primary rat pulmonary arterial smooth muscle cells (PASMCs) were divided into 4 groups: a normoxia control group, a normoxia+MDL28170 group, a hypoxia group and a hypoxia+MDL28170 group. Cell proliferation was detected by MTS and flow cytometry. Levels of Ki-67 and proliferating cell nuclear antigen (PCNA) mRNA were determined by real-time PCR.
 RESULTS: RVSP, mPAP and right ventricular remodeling index were significantly elevated in the hypoxia group compared to those in the normoxia group. In the hypoxia group, pulmonary vascular remodeling was significantly developed, accompanied by up-regulation of calpain-1, -2 and -4. MDL28170 significantly inhibited hypoxia-induced proliferation of PASMCs concomitant with the suppression of Ki-67 and PCNA mRNA expression.
 CONCLUSION Calpain mediates vascular remodeling via promoting proliferation of PASMCs in hypoxia-induced pulmonary hypertension.
Animals ; Calpain ; genetics ; physiology ; Cell Proliferation ; Dipeptides ; physiology ; Hypertension, Pulmonary ; chemically induced ; genetics ; physiopathology ; Hypertrophy, Right Ventricular ; Hypoxia ; Ki-67 Antigen ; drug effects ; Myocytes, Smooth Muscle ; physiology ; Proliferating Cell Nuclear Antigen ; drug effects ; Pulmonary Artery ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Up-Regulation ; Vascular Remodeling ; genetics ; physiology

OBJECTIVETo develop an HPLC method for fingerprint study of both Senecio scandens and S. scandens.

METHODFingerprints of the two Senecio herbs were compared. And the concentrations of main peaks in them were semi-quantified as chlorogenic acid or hyperoside. Chromatography was performed on a Shiseido Capcell Pak MG II C18 (4.6 mm x 250 mm, 5 microm) column using a gradient elution of acetonitrile-water containing 0.2% acetic acid. And detection wavelength was 360 nm.

RESULTSignificant difference was found in the fingerprint of the two herbs. Eleven peaks were picked out for the evaluation of S. scandens and S. scandens, respectively. They were identified to be either organic acid compounds or flavones by HPLC-UV and LC-ESI-MS analysis. And semi-quantification of them showed the concentrations of organic acid compounds and flavones in S. scandens were 2.29- and 15.56- folds of those in S. scandens, respectively.

CONCLUSIONThe developed HPLC method is suitable for the fingerprint study for both of S. scandens and S. scandens. It is robust and producible enough to be used for the quality evaluation on S. scandens.

Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; methods ; Quercetin ; analogs & derivatives ; analysis ; Senecio ; chemistry

OBJECTIVETo develop an UPLC-MS method for the simultaneously quantitation of adonifoline and senecinine in Senecio herbs.

METHODUPLC-Micro 2000 was used for quantification in SIR mode under ESI+. Monocrotaline was used as the internal standard. Chromatography was performed on a Shiseido Capcell Pak MG (2.0 mm x 50 mm, 3 microm) column at 30 degrees C using an gradient elution of acetonitrile-0.5% formic acid in water at the flow rate of 0.3 mL x min(-1). The injection volume was 2 microL.

RESULTGood linearity was obtained for quantitation of adonifoline over the range of 1.02-816.00 microg x L(-1) (r = 0.998 0). And recoveries at different concentration levels were between 95.73% and 103.0% with RSDs no more than 2.5%. For quantification of senecionine, the linear range was between 1.08-860.56 microg x L(-1) (r = 0.997 6). And recoveries at different concentration levels were between 95.67% and 101.5% with RSDs no more than 2.3%. Good reproducibility and precision were also achieved.

CONCLUSIONThe new developed UPLC method is sensitive, accurate and reliable enough for the quantitation of adonifoline and senecionine in Senecio herbs thus can be used for the limit detection of pyrrolizidne alkaloids in S. scandens. It can also be used for the identification of fake drugs of S. scandens such as S. vulgaris. The developed method was served for the quality evaluation of Herba Senecionis Scandentis.

Chromatography, Liquid ; methods ; Lactones ; analysis ; Mass Spectrometry ; methods ; Pyrrolizidine Alkaloids ; analysis ; Senecio ; chemistry

OBJECTIVETo study the anti-platelet effect and influence of Radix Paeoniae Rubra and Radix Paeoniae Alba on rat's endogenous metabolites by animal experiment and UPLC-MS based metabolomic method.

METHODAfter administration of 80% ethanol extracts of Radix Paeoniae Rubra and Radix Paeoniae Alba for 6 d, the serum samples were analyzed by ultra-performance liquid chromatography- mass spectrometry (UPLC-MS) and partial least squares-discriminant analysis (PLS-DA) to find out the potential biomarker.

RESULTBoth of the extracts of Radix Paeoniae Rubra and Radix Paeoniae Alba have good effects of inhibition on platelet coacervation, and the effect of Radix Paeoniae Alba is better than that of Radix Paeoniae Rubra. Malic acid, alpha-acetone dicarboxylic acid, leukotrieneA4 (LTA4), prostaglandinE2 (PGE2) and prostaglandin F2alpha (PGF2alpha) are proved to be significant expressed biomarkers.

CONCLUSIONMetabolomics is helpful for the further research of the mechanism of anti-platelet action of Radix Paeoniae Rubra and Radix Paeoniae Alba.

Animals ; Chromatography, Liquid ; methods ; Least-Squares Analysis ; Male ; Mass Spectrometry ; methods ; Metabolomics ; Paeonia ; Plant Extracts ; pharmacology ; Platelet Aggregation Inhibitors ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo compare the anti-inflammatory effect of Radix Paeoniae Rubra and Radix Paeoniae Alba by animal experiment and metabolimic analysis.

METHODTo establish the rats model of toes swelling caused by carrageenan, study the anti-inflammatory effect of Radix Paeoniae Rubra and Radix Paeoniae Alba. The serum samples were analyzed by ultraperformance liquid chromatography-mass spectrometry (UPLC-MS), to find out the potential identification biomarker by PLS-DA.

RESULTBoth of the extracts of Radix Paeoniae Rubra and Radix Paeoniae Alba have good effects of inhibition to swelling caused by carrageenan in 0.5-1 h, and the extract of Radix Paeoniae Rubra also show significant inhibition in 2-3 h. Glutathione( GSH), gamma-aminobutyric acid (GABA), prostaglandin F2alpha (PGF2alpha), prostaglandinE3 (PGE3), leukotrieneA4 (LTA4), prostaglandinE2 ( PGE2) are proven to be significant expressed biomarkers. Radix Paeoniae Rubra and Radix Paeoniae Alba may have great influence on PGF2alpha and PGE3. There was also significant difference between the effects of Radix Paeoniae Rubra and Radix Paeoniae Alba, which suggested the difference of anti-inflammatory between the two herbs.

CONCLUSIONThe results of metabolomics are related with the results of classic pharmaco- experiment, which is helpful for the further research of the mechanism of action of Radix Paeoniae Rubra and Radix Paeoniae Alba.

Alprostadil ; analogs & derivatives ; blood ; Animals ; Anti-Inflammatory Agents ; pharmacology ; Chromatography, Liquid ; methods ; Dinoprost ; blood ; Male ; Metabolomics ; Paeonia ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Mass, Electrospray Ionization ; methods