To develop a LC-MS/MS method for the determination of protocatechuic acid, protocatechuic aldehyde, salvianolic acid A, salvianolic acid B, cryptotanshinone and tanshinone II(A) in rat plasma and brain. The plasma and brain samples were precipitated with ethyl acetate, then were separated on an Agilent eclipse plus-C18 column (2.1 mm x 50 mm, 3.5 microm) using acetonitrile (consisting of 0.1% formic acid) and water (consisting of 0.1% formic acid) as mobile phase in gradient elution mode. The mass spectrometer was operated under both positive and negative ion mode with the ESI source, and the detection was performed by MRM. The transition of 154.3/153.1 m/z for protocatechuic acid, 137.3/108 m/z for protocatechuic aldehyde, 493.0/295.2 m/z for Salvianolic acid A, 718.0/520.0 m/z for salvianolic acid B, 321.4/152.3 m/z for chloramphenicol, 297.4/254.3 m/z for cryptotanshinone, 295.5/249.3 m/z for tanshinone II(A) and 285.2/154.0 m/z for Diazepam. The calibration curves in the range of 0.625-1 000 microg x L(-1) for protocatechuic acid and protocatechuic aldehyde, 1.25-1 000 microg x L(-1) for salvianolic acid A, 2.5-1 000 microg x L(-1) for salvianolic acid B, 0.15-1 000 microg x L(-1) for cryptotanshinone, 0.625-1 000 microg x L(-1) for tanshinone II(A) are with good linearityin rat plasma and brain. The analysis method is sensitive, simple, and suitable enough to be applied in the pharmacokinetic study of the 6 main components. Animal testing gives the lgBB of the drugs and further studies of the 6 components cross the blood-brain barrier can be carried out.
Animals ; Benzaldehydes ; administration & dosage ; blood ; pharmacokinetics ; Benzofurans ; administration & dosage ; blood ; pharmacokinetics ; Blood-Brain Barrier ; metabolism ; Brain ; metabolism ; Caffeic Acids ; administration & dosage ; blood ; pharmacokinetics ; Catechols ; administration & dosage ; blood ; pharmacokinetics ; Chromatography, Liquid ; methods ; Diterpenes, Abietane ; administration & dosage ; blood ; pharmacokinetics ; Hydroxybenzoates ; administration & dosage ; blood ; pharmacokinetics ; Injections, Intravenous ; Lactates ; administration & dosage ; blood ; pharmacokinetics ; Phenanthrenes ; administration & dosage ; blood ; pharmacokinetics ; Plant Preparations ; administration & dosage ; blood ; pharmacokinetics ; Rats ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry ; Tandem Mass Spectrometry ; methods

Animals ; Blood Coagulation ; drug effects ; Blood Platelets ; drug effects ; Female ; Haplorhini ; Humans ; Interleukin-11 ; pharmacology ; Male ; Platelet Count ; Thrombopoietin ; pharmacology

Objective To explore the prevalence of ETS gene fusion in prostate cancer and its correlation with patient's clinicopathologic index.Methods Ninety-one samples from prostate needle biopsy cases with median age 75 (55-90) years and 18 samples from radical prostatectomy cases with median age 72 (63-81) years were collected from Oct.2010 to Feb.2012.TMPRSS2-ERG,TMPRSS2-ETV1 and TMPRSS2-ETV4 fusions were tested by multi-probe fluorescence in situ hybridization (FISH) assay.Fusion positive and negative cases were compared with age,TNM stage,Gleason score (median Gleason score 7 (6-9)),and PSA value (median PSA 33.4 μg/L,mcan PSA 118.8 μg/L).In needle biopsy cases,there were early stage 32 (34％),locally advanced prostate cancer 36 (40％),metastasis 23 (26％) ; in radical prostatectomy cases,there were 9 cases in T2 stage,8 cases in T3 stage,1 cases in T4a stage,respectively.Results TMPRSS2-ERG fusions were present in 14.3％ (13/91,95％ confidence interval,0.071-0.215)biopsy specimens and in 11.1％ (2/18,95％ confidence interval,0-0.256) radical prostatectomy specimens.No TMPRSS2-ETV1 or TMPRSS2-ETV4 fusions were found in any cases.Altogether,13 (86.7％)cases possessed deletion pattern.And 7 (46.6％) hold insertion pattern.5 cases had both deletion and insertion pattern.38.5％ (5/13) deletion pattern had distant metastasis.Except one metastatic case harbored both deletion and insertion pattern,there was no insertion pattern accompanied with metastasis.There were no differences between fusion positive and negative cases in the distribution of age (P =1.000),PSA (P =1.138),primary Gleason score (P=2.186),Gleason score (P=2.107),TNM stage (P=2.052) and Risk Degree (P=2.597).Conclusions The TMPRSS2-ERG fusion positive cases harbor more deletion pattern than insertion pattern.There are no differences between fusion positive and negative cases in the distribution of age,PSA,Gleason score,TNM stage and Risk Degree.

OBJECTIVETo compare the clinical safety and efficacy of laparoscopic versus open colorectal resection in octogenarians. Methods Studies comparing laparoscopic colorectal resection with open colorectal resection in octogenarians were identified from the Medline, Embase, Ovid, and Cochrane databases from 1990 to 2012. The methodological quality of the selected studies was assessed to determine studies suitable for inclusion. Meta-analysis was performed by fixed or random effects model.

RESULTSFive observational studies with a total of 685 patients (330 laparoscopic colorectal resections and 355 open colorectal resections) were identified. Laparoscopic colorectal resection was associated with a prolonged operative time (WMD=27.89, P<0.01) and a lower rate of overall complications (OR=0.58, P<0.01), wound infection (OR=0.50, P<0.05), cardiovascular complication(OR=0.53, P<0.05), quicker bowel function return (WMD=-0.83, P<0.01), and shorter length of hospital stay (WMD=-3.60, P<0.05). No differences were found with regard to anastomotic leak (OR=1.13, P>0.05), prolonged ileus (OR=0.71, P>0.05), respiratory complication (OR=0.59, P>0.05),mortality (OR=0.67, P>0.05), and reoperation (OR=0.85, P>0.05).

CONCLUSIONLaparoscopic colorectal resection is as safe as open colorectal resection, and is more favorable in terms of length of hospital stay and bowel function return in octogenarians.

Adult ; Chemical and Drug Induced Liver Injury ; Dimethylformamide ; poisoning ; Humans ; Male

The combined application of statins that inhibit HMG-CoA reductase and fibrates that activate PPAR-α can produce a better lipid-lowering effect than the simple application, but with stronger adverse reactions at the same time. In the treatment of hyperlipidemia, the combined administration of TCMs and HMG-CoA reductase inhibitor in treating hyperlipidemia shows stable efficacy and less adverse reactions, and provides a new option for the combined application of drugs. In this article, the pharmacophore technology was used to search chemical components of TCMs, trace their source herbs, and determine the potential common TCMs that could activate PPAR-α. Because there is no hyperlipidemia-related medication reference in modern TCM classics, to ensure the high safety and efficacy of all selected TCMs, we selected TCMs that are proved to be combined with statins in the World Traditional/Natural Medicine Patent Database, analyzed corresponding drugs in pharmacophore results based on that, and finally obtained common TCMs that can be applied in PPAR-α and combined with statins. Specifically, the pharmacophore model was based on eight receptor-ligand complexes of PPAR-α. The Receptor-Ligand Pharmacophore Generation module in the DS program was used to build the model, optimize with the Screen Library module, and get the best sub-pharmacophore, which consisted of two hydrogen bond acceptor, three hydrophobic groups and 19 excluded volumes, with the identification effectiveness index value N of 2. 82 and the comprehensive evaluation index CAI value of 1. 84. The model was used to screen the TCMD database, hit 5,235 kinds of chemical components and 1 193 natural animals and plants, and finally determine 62 TCMs. Through patent retrieval, we found 38 TCMs; After comparing with the virtual screening results, we finally got seven TCMs.
Acyl Coenzyme A ; metabolism ; Animals ; Databases, Factual ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Hydrophobic and Hydrophilic Interactions ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; chemistry ; pharmacology ; Lipid Metabolism ; drug effects ; Lipids ; blood ; Medicine, Chinese Traditional ; Models, Molecular ; Quantitative Structure-Activity Relationship ; Technology

Inflammatory bowel disease (IBD) has become a common ailment in China in the past decade. At present, the level of diagnosis and treatment of inflammatory bowel disease in China still falls behind Western countries, while the incidence increases rapidly. Chinese IBD Working Group made a new consensus on diagnosis and treatment of inflammatory bowel disease in 2012. This article is to interpret some key issues in the consensus.
China ; Humans ; Inflammatory Bowel Diseases ; diagnosis ; therapy

Objective To investigate the protective effects of the 14-3-3 protein overexpression on the injury of PC12 cell induced by MPP~+ and its mechanisms.Methods For expression in mammalial cells, pcDNA3.1(+)-14-3-3 plasmid was constructed and transfeeted into PC12 cell with Lipofectamine~(TM)2000. The overexpression of transfected 14-3-3 gene in PC12 cell was determined by immunofluorescence and Western blotting.The effects of 14-3-3 overexpressing on the cells viability,apoptotie ratio and the activity of superoxide dismutase(SOD)as well as glutathione peroxidase(GSH-Px)of PC 12 cell treated with MPP~+ were measured by MTT assay,flow cytometry analysis and microplate reader respectively.Results The expression of 14-3-3 protein in transfection group(1.19?0.06)increased evidently compared with control group(0.75?0.05).And the antioxidant enzyme activity assession,MTT assay and flow cytometry analysis shows that the overexpression of 14-3-3 protein elevates the activity of SOD(transfection group:(9.13? 0.41)U/mg protein,MPP~+ group:(6.45?0.52)U/mg protein)and GSH-Px(transfection group: (89.66?3.42)?mol/mg,protein MPP~+ group:(82.73?4.15)?mol/mg protein),increases the cell viability(transfection group:0.78?0.06,MPP~+ group:0.54?0.07),and inhibits cell apoptosis (transfeetion group:11.87%?3.26%,MPP~+ group:36.30%?2.39%)of PC12 induced by MPP~. Conclusion The overexpression of 14-3-3 protein could elevate the activity of antioxidant enzymes SOD and GSH-Px,reduce oxidant stress,alleviate MPP~+ toxicity,and thus inhibit the apoptosis of PC12 cell induced by MPP~+.

Objective To evaluate the expression of alpha-methylacyl-coenzyme-A racemase (AMACR,P504s) with immunohistochemical staining in diagnosis of prostate carcinoma. Methods A total of 133 specimens were taken from the patients (mean age,71 years),including 46 cases of prostate carcinoma (PCa) (1 case of stage A,19 cases of stage B,14 cases of stage C,12 cases of stage D;4 cases of gradeⅠ,14 cases of gradeⅡ,28 cases of grade Ⅲ),53 cases of benign prostate hyperplasia (BPH),13 cases of prostate intraepithelial tumor (PIN),9 cases of normal prostate,6 cases of prostatitis,3 cases of metastatic prostate cancer.Two sections of each specimen were made,with 1 stained by hematoxylin and eosin,and the other stained immunohistochemically by a rabbit monoclonal antibody to AMACR (P504s).AMACR staining expression was characterized as negative (score,1), weak (positive) (2), moderate (3) or strong (4). Results In 46 cases of PCa,AMACR staining expression was negative in 2 cases,weak in 1,moderate in 25 and strong in 18,with a mean staining intensity of 3.28 [95% confidence interval (CI), 3.07-3.50]. In 53 cases of BPH, the staining expression was negative in 47 cases, weak in 6, with a mean intensity of 1.11 (95% CI, 1.02-1.20).In 13 cases of PIN, the staining expression was negative in 12, weak in 1, with a mean intensity of 1.08 (95% CI, 0.91-1.24).The score of PCa group was significantly different from those of the latter 2 groups (P

Objective To investigate the low-dose hyper-radiosensitivity (HRS)/induced radioresistance (IRR) in A549 cells synchronized at G2 phase and the role of ATM kinase in the process.Methods Human lung adenocarcinoma cell line A549 was synchronized at G2 phase by aphidicolin.The ATM-specific activator and inhibitor,chloroquine and KU55933,were used to regulate the activity of ATM.The colony formation assay was used to evaluate cell survival.Flow cytometry was used to determine the cell cycle of radiation-exposed A549 cells synchronized at G2 phase.Immunofluorescence was used to observe the dynamics of γ-H2AX fluorescence and evaluate the efficiency of DNA repair in A549 cells synchronized at G2 phase.Western blot was used to detect the expression of phosphorylated ATM (Ser1981) and ATM.Results A549 cells synchronized at G2 phase had substantially enhanced HRS than non-synchronized cells.The dose-induced transition from HRS to 1RR was in accordance with the dose-response pattern of early G2/M checkpoint.However,with the same threshold dose,the activation of early checkpoint occurred earlier and lasted longer than normal.The activation of ATM kinase inhibited HRS and enhanced DNA repair,while the inhibition of ATM kinase enhanced HRS and hindered DNA repair.Conclusions ATM kinase-mediated early G2+M checkpoint is a molecular switch for HRS in synchronized A549 cells.Low-dose irradiation with G2-phase synchronization and ATM inhibitor can enhance the low-dose radiosensitivity.