The study aims to establish a method for simultaneous determination of repaglinide and pravastatin sodium in rat plasma by LC-MS/MS and to study its pharmacokinetic interactions. Eighteen male SD rats were divided into repaglinide group, pravastatin sodium group and co-administration group. Blood samples were collected at different times after oral administration. Repaglinide and pravastatin sodium in rat plasma were separated by Agilent HC-C18 with the mobile phase consisting of methanol-0.1% formic acid (80 : 20). Detection and quantification were performed by using ESI-MS. The detector was operated in selected Reaction-monitoring mode at m/z 453.3-->230.1 for repaglinide, m/z 447.2-->327.4 for pravastatin sodium and m/z 285.1-->192.9 for diazepam as the internal standard. The calibration curve obtained was linear (R2>0.99) over the concentration range of 9.77-10,000 ng.mL-1 for repaglinide and 4.88-625 ng.mL-1 for pravastatin sodium. Compared with the single administration group, Cmax and AUC0-6h of repaglinide increased significantly (P<0.05) and tmax of pravastatin sodium prolonged (P<0.05) in co-administration group. The method is found to be simple, sensitive and accurate for determining the concentration of repaglinide and pravastatin sodium in rat plasma. There exists pharmacokinetic interactions in the co-administration of repaglinide and pravastatin sodium.
Administration, Oral ; Animals ; Carbamates ; administration & dosage ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; Drug Interactions ; Male ; Piperidines ; administration & dosage ; blood ; pharmacokinetics ; Pravastatin ; administration & dosage ; blood ; pharmacokinetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Sensitivity and Specificity ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

This study aims to investigate the change of plasma concentration of digoxin (DIG) in rats with ovariectomy. Twelve female SD rats were randomly assigned into ovariectomized group and sham group (n = 6). All rats plasma was collected after a single dose of 2 mg x kg(-1) DIG administrated orally, serum DIG concentration was determined by LC-MS/MS. The level of P-gp in the intestinal was analyzed by Western blotting. Pharmacokinetic calculations were performed on each individual using DAS 2.0 practical pharmacokinetic software. Compared with the sham group, C(max) of ovariectomized group decreased significantly (P < 0.01). There was no significant difference of AUC(0-t), and the level of P-gp was elevated in ovariectomized group. It was found that C(max) of DIG was significantly reduced after ovariectomy, and the change was associated with the decreased level of estrogen, which contributes to the increased level of P-gp.
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Animals ; Blotting, Western ; Chromatography, Liquid ; Digoxin ; blood ; pharmacokinetics ; Disease Models, Animal ; Estrogens ; blood ; Female ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

Adenoma ; genetics ; metabolism ; pathology ; surgery ; Adult ; Carcinoma, Renal Cell ; genetics ; metabolism ; pathology ; Chromosomes, Human, Pair 3 ; genetics ; Chromosomes, Human, Pair 7 ; genetics ; Diagnosis, Differential ; Diploidy ; Female ; Follow-Up Studies ; Humans ; Kidney Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Retrospective Studies ; S100 Proteins ; metabolism ; Vimentin ; metabolism ; WT1 Proteins ; metabolism ; Wilms Tumor ; metabolism ; pathology

The tetranucleotide repeat locus D7S809 was investigated in Cantonese population by polymerase chain reaction (PCR) amplification, subsequent polyacryramide gel electrophoresis and silver staining. 14 alleles and 50 genotypes were detected in 190 sample individuals. All alleles differed in size by 4 bp repeat. No interalleles were found. The heterozygosity, power of discrimination and chance of non-paternal exclusion were 0.8613, 0.9645 and 0.7184 respectively. No significant deviations from Hardy-Weinberg equilibrium were found. The D7S809 was a highly polymorphic, stable and accurate type locus and had been successfully applied to 100 paternity test cases.
Alleles ; Asian People/genetics* ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Microsatellite Repeats/genetics* ; Paternity ; Polymorphism, Genetic

OBJECTIVETo study the effect of hypoxia-inducible factor-1α (HIF-1α) on human gastric cancer cells apoptosis in simulated CO2 pneumoperitoneum environment.

METHODSApplied closed box to simulated CO2 pneumoperitoneum environment under the pressure of 0, 5, 10 and 15 mm Hg (1 mm Hg = 0.133 kPa). Compared HIF-1α mRNA and protein expression of MKN-45 cells before and after silencing HIF-1α by RT-PCR and Western blot. Study the changes of bcl-2/bax expression in MKN-45 cells before and after silencing HIF-1α by immunohistochemistry. The apoptosis ratio of MKN-45 was measured by using Annexin V-FITC/PI double labelled staining.

RESULTSIn 15 mm Hg group, HIF-1α mRNA and protein expression of MKN-45 cells (1.48 ± 0.22, 1.34 ± 0.09) and HIF-1α protein expression in 10 mm Hg group (1.25 ± 0.10) were significantly higher than those in control group (0.55 ± 0.17, 0.83 ± 0.04) (P < 0.05). But there was no significant differences among 0, 5, 10 mm Hg group and control group in HIF-1α mRNA (P > 0.05); and no obvious difference was found among 0, 5 mm Hg group and the control group in HIF-1α protein expression (P > 0.05). In 15 mm Hg CO2 pressure, bcl-2/bax expression (0.78 ± 0.05) was obviously lower than that in the control group (1.43 ± 0.15) (P < 0.05) and the apoptosis ratio (11.70 ± 0.12) was significantly higher than the control group (0.22 ± 0.07) (P < 0.01) before silencing HIF-1α. But once HIF-1α was silenced, HIF-1α mRNA (0.52 ± 0.11), HIF-1α protein expression (0.92 ± 0.02), bcl-2/bax ratio (1.57 ± 0.04) and apoptosis ratio (0.45 ± 0.11) in MKN-45 were not significantly different between 15 mm Hg group and the control group (P > 0.05).

CONCLUSIONSThe apoptosis ratios of MKN-45 under 0, 5, 10 mm Hg CO2 pneumoperitoneum are comparable with that in the control group before the silencing of HIF-1α. The apoptosis ratio of MKN-45 is increased under 15 mm Hg CO2 pneumoperitoneum environment and HIF-1α may be one of the important factor to improve the apoptosis of human gastric cancer cell.

Apoptosis ; Carbon Dioxide ; physiology ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Pneumoperitoneum, Artificial ; RNA Interference ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVE:To explore an effective method to formulate management-related strategies for off-lable use of drugs by the evidence-based medicine. METHODS:The process of guideline formulation included seven procedures,i.g. establishment ofguideliesformulation workgroup;investigation and selection of the status quo on off-label drug use;identification of the clinical problems;retrieval and evaluation and comprehensing of evidence;applification of GRADE in evidence quality grading;formation of the recommendations consensus;peer review and result publication. And eventually guidelines were formed based on the steps. This study took off-label use of rheumatoid immunoprotective subjects as a case to explore. RESULTS & CONCLUSIONS:Based on the evidence evaluation system and above 7 steps,the methods and process of guideline formulation on off-label use of rheuma-toid immunoprotective subjects that integrated administration,law,clinical medicine,pharmacy subjects were made .The process of guideline formulation fully reflects multidisciplinary characteristics of the workgroup,the advanced nature of the process,the comprehensiveness of evidence ,the rigor of evidence quality grading,and the normalization of consensus. It provides reference in methodology for establishing a comprehensive evidence-based evaluation and management system of off-label use of drugs for all clinical specialist disease. Therefore,this scientific research results may promote the standardization and legalization of the off-label use of drugs management in China.

Department of General practice of West China Hospital of Sichuan University has been commissioned to train general practitioner for Tibet Autonomous Region since 2015. Based on the "demand-oriented and clinical ability training", the established training framework includes four aspects: a rotation plan of general practice theory-clinical practice-community application, the closed-loop mechanism of teaching management system improvement,innovation of training assessment and teaching activities. This article summarizes the practical experience of the commissioned general practice residency training for Tibet to provide reference for the program design.

Objective To investigate the effect of bile acids on altered synthesis and transport in the pathogenesis of colitis-related cholestasis and the underlying mechanisms in rats.Methods Adult Wistar male rats were randomly divided into normal group and model group,ulcerative colitis(UC) model was established by administration 2,4,6-trinitro-benzenesulfonicacid in intro-colonic,while rats in normal group were colonic administered with saline.UC model was evaluated by calculating disease activity index,histopathological score and myeloperoxidase activity.Serum level of alkaline phosphatase (ALP),glutamyl transpeptidase (GGT),alanine aminotransferase (ALT) and aspartate aminotransferase(AST) were also determined.Moreover,the level of hepatic total bile acids,deoxycholic acid and lithocholic acid were determined by HPLC/MS.The hepatic protein expression of bile acids synthesis enzyme cholesterol 7α-hydroxylase(Cyp7a1) as well as bile acids export transporter multidrug resistance-associated protein 2 (Mrp2) were all investigated by Western-bolt.Results Compared to the normal group,disease activity index,histopathological score and myeloperoxidase activity in model group were all significantly increased to (4.51 ± 1.49),(1.36 ± 0.69) point,and (0.40 ± 0.07) U · mg-1,respectively (all P ＜ 0.05),suggesting the successful preparation of UC rat model.Compared to the normal group,the content of total bile acids and deoxycholic acid were all markedly increased to (85.50 ± 18.60) μmol · L-1 and (1.50 ± 0.68) ng · g-1 in model group,and the serum level of ALP and GGT were all significantly increased to (259.43 ±58.58) U · L-1and (1.50 ±0.68) U · L-1 in model group (all P ＜ 0.05),indicating the occurrence of cholestasis in model group of rats.Compared to the normal group,the protein expression of Cyp7a1 in the liver of model group was significantly increased to 0.72 ± 0.07 in gray value,but the protein expression of Mrp2 was markedly decreased to 0.66 ± 0.04 in gray value in model group (all P ＜ 0.05).Conclusion Acute colitis can induce intrahepatic cholestasis in rats,increased bile acids synthesis and decreased bile acids excretion may contribute to colitis-related cholestasis via up-regulating Cyp7a1 expression and down-regulating Mrp2 expression in the liver of colitis rats.

AIM:To investigate the changes of cardiac structure and function in rats with type 2 diabetic melli-tus(T2DM),and to explore the mechanisms underlying diabetic cardiomyopathy.METHODS:The cardiac structure and function were measured by echocardiography in Zucker diabetic fatty(ZDF)rats and their control Zucker lean(ZL)rats. The size of the cardiomyocytes was determined by wheat germ agglutinin staining.The protein expression of atrial natriuretic peptide(ANP),β-myosin heavy chain(β-MHC), receptor for advanced glycation end products(RAGE), L-type cal-cium channel α1C subunit(CaV1.2)and Orai1 was assessed by Western blot.RESULTS:Compared with the ZL control rats,the thickness of left ventricular wall,ejection fraction(EF),fractional shortening(FS)and the sizes of cardiomyo-cytes were significantly increased,and diastolic function was decreased in the ZDF rats(P<0.05).The protein expression of β-MHC, ANP, RAGE and Orai1 was increased, while the expression of Ca V1.2 was decreased in ZDF rats(P <0.05).CONCLUSION:T2DM rats show the prominent features including cardiomyocyte hypertrophy,ventricular hyper-trophy and compensatory enhancement of cardiac function, and the Ca2+handling and increase in RAGE expression may play important roles in the processes.

Background:The Chinese appropriate use criteria (AUC) for coronary revascularization was released in 2016 to improve the use of coronary revascularization.This study aimed to evaluate the association between the appropriateness of coronary revascularization based on the Chinese AUC and 1-year outcomes in stable coronary artery disease (CAD) patients.Methods:We conducted a prospective,multi-center cohort study of stable CAD patients with coronary lesion stenosis ≥50％.After the classification of appropriateness based on Chinese AUC,patients were categorized into the coronary revascularization group or the medical therapy group based on treatment received.The primary outcome was a composite of death,myocardial infarction,stroke,repeated revascularization,and ischemic symptoms with hospital admission.Results:From August 2016 to August 2017,6085 patients were consecutively enrolled.Coronary revascularization was associated with a lower adjusted hazard of 1-year major adverse cardiovascular and cerebrovascular events (MACCEs;hazard ratio [HR]:0.62;95％ confidence interval [CI]:0.45-0.86;P =0.004) than medical therapy in patients with appropriate indications (n =1617).No significant benefit in 1-year MACCEs was found after revascularization compared to after medical therapy in patients with uncertain indications (n =2658,HR:0.81;95％ CI:0.52-1.25;P =0.338) and inappropriate indications (n =1810,HR:0.80;95％ CI:0.51-1.23;P=0.308).Contusions:In patients with appropriate indications according to Chinese AUC,coronary revascularization was associated with significantly lower risk of MACCEs at 1 year.No benefit was found in coronary revascularization in patients with inappropriate indications.Our findings provide evidence for using Chinese AUC to guide clinical decision-making.