A rapid and sensitive liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) method for quantification of sitagliptin in human plasma and urine had been developed. This method was applied to the pharmacokinetics study of sitagliptin tablet after single- and multiple-dosing in Chinese population. Plasma samples were prepared by a liquid-liquid extracted method, and urine samples were diluted. Compounds were analyzed by multiple reaction monitoring (MRM) mode with a electrospray ionization (ESI) interface. Mobile phase consisted of methanol and water (85 : 15, v/v). The linear concentration range of calibration curve was 0.5-1 000 ng.mL-1. and 0.2-100 µg.mL , intra-run/between-run accuracy was 98.98%-103.69% and 97.63%-102.29%, intra-run/between-run precision was <5.51% and 4.26% for plasma and urine sample, respectively. The stability of sitagliptin stock solution was tested for 55 days at -30 °C. Sitagliptin was stable when stored under the following conditions: 24 hours in the autosampler after sample preparation; 24 hours at room temperature, after 3 freeze and thaw cycles (from -30 °C to room temperature), 40 days at -30 °C for plasma and urine samples. The absolute recovery in plasma was 71.1%, and no matrix effect was founded. This method was proved simple, specific, sensitive, rapid and suitable for pharmacokinetics study of sitagliptin in human being.
Calibration ; Chromatography, Liquid ; Humans ; Liquid-Liquid Extraction ; Sitagliptin Phosphate ; blood ; pharmacokinetics ; urine ; Tandem Mass Spectrometry

Objective To compare the value of speckle-tracking imaging and tissue Doppler strain imaging in evaluating left atrial (LA) mechanical contraction. Methods Forty patients after successful percutaneous atrial septal defect (ASD) closure were enrolled in this study. Two-dimensional strain imaging (2D strain echocardiography, 2DSE) and tissue Doppler imaging (TDI) were performed one weak after ASD closure. Off-line analysis was done for atrial longitudinal peak systolic strain on the interatrial septum, in correspondence of the device, and on the lateral wall of the left atrium from apical 4 chamber view with 2DSE and color-coded strain imaging, respectively. Results There was no statistical difference of the inter- and intra-observer viability as well as time consumption on the off-line analysis between 2DSE and TDI. LA lateral wall strain was much higher than that of LA septum assessed by both color-coded strain imaging (47.31%±27.25% vs 30.06%±14.29%, P<0.01) and 2DSE (43.49%±25.55% vs 12.74%±9.16%, P<0.001). LA lateral wall strain was similar with either method (P=NS). However, at the site of ASD occluder, a non-contractile element, deformation was significantly higher in color-coded strain imaging than 2DSE (P<0.001). Five patients (12.50%) presented absence of deformation on ASD occluder with 2DSE, whereas there was no patient with TDI assessment (χ~2=5.33, P=0.027). Conclusion Speckle-tracking imaging is superior to tissue Doppler strain imaging in assessing left atrium mechanical function.

Animals ; Dementia, Vascular ; metabolism ; physiopathology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Hippocampus ; cytology ; metabolism ; physiopathology ; Male ; Mice ; Neurons ; metabolism ; physiology ; Random Allocation

Abstract?AIM:To evaluate the pupil light reflex in patients with primary open angle glaucoma, and to investigate the relation between pupil and visual field defect.?METHODS:From July 2014 to October 2015, 115 eyes in 86 patients with primary open angle glaucoma and 23 eyes in 16 healthy individuals were continuously enrolled in this study.All the subjects received comprehensive eye examination, visual field examination ( Humphrey, SITA Standard 24-2 ) and dynamic pupil measurement ( MonCV3 Metrovision) .According to the visual field and the Glaucoma Staging System, the patients with glaucoma were divided into 5 subgroups: stage 1, stage 2, stage 3, stage 4 and stage 5. The parameters of pupillary light reflex were as follows: pupil diameter ( minimum, maximum ) , latency and duration of contraction, latency and duration of dilatation, contraction amplitude, contraction and dilatation speed, and percent of pupil contraction ( PPC ). SPSS 19.0 statistical software was used to analyze the measurement results.?RESULTS:The control group significantly differed from the stage 4 subgroup ( P=0.032 ) and stage 5 subgroup (P=0.014) in terms of minimum pupil diameter; there was significant difference in the pupil contraction speed between groups ( F =648.675, P <0.01 ), and the contraction speed in stage 5 subgroup was significantly lower than those in the other subgroups and control group (P<0.05); the control group significantly differed from the stage 3, stage 4, and stage 5 subgroup in terms of PPC ( P<0.05 ).Pupil contraction speed, PPC and minimum diameter showed correlation with the stages of glaucoma.?CONCLUSION:Pupil contraction ability in patients with primary open angle glaucoma was impaired, and the degree of impairment is related with the degree of visual field defect.

By functional plates,16 strains which can produce?-mannana-se were isolated frnm 28 Bacillus spp.Using a pair of degenerated primers,the conserved fragments of?-mannanase gene from the selected strains were amplified by PCR.The obtained nucleotide fragments were sequenced and compared with the homologous?-mannanase genes in GenBank and a phylogenetic tree was generated.Comparing to the genes coding?-mannanase published,the cloned nucleotide fragments show the highest sequence identity between 62% and 98%.The genes coding fnr?-mannanase of Bacillus circulus have low identity while the?-mannanase genes of Bacillus subtilis and other Bacillus spp. have high identity.

Adult ; Aged ; Aged, 80 and over ; Benign Paroxysmal Positional Vertigo ; diagnosis ; drug therapy ; economics ; Female ; Humans ; Male ; Middle Aged ; Surveys and Questionnaires ; Treatment Outcome

Ataxia Telangiectasia Mutated Proteins ; Breast ; metabolism ; pathology ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; pathology ; Cell Cycle Proteins ; metabolism ; Checkpoint Kinase 2 ; DNA-Binding Proteins ; metabolism ; Female ; Humans ; Lymphatic Metastasis ; Neoplasm Grading ; Protein-Serine-Threonine Kinases ; metabolism ; Tumor Burden ; Tumor Suppressor Protein p53 ; metabolism ; Tumor Suppressor Proteins ; metabolism

Animals ; Hydroxy Acids ; toxicity ; Lacquer ; toxicity ; Mice ; Mice, Inbred ICR ; Superoxide Dismutase ; metabolism

Objective To establish a method for detecting HBV cccDNA in hepatocytes of chronic hepatitis B patients.Method 21 liver biopsies from the hepatic operation patients in the hospital of jiangsu province,concluding 19 HBV chronic infected patients (10 HBeAg positive patients and 9 HBeAg negative patients) and 4 uninfected patients,HBV DNA(+) serum of hepatitis B patients was thought as rcDNA.To use proteinase K to release HBV cccDNA and genomic DNA,then divide the cell lysis solution into two parts,one for detecting HBV cccDNA,the other for detecting the number of ?-Globin as internal control. Nucleic acid for detecting HBV cccDNA extracted by phenol-chloroform was digested by plasmid-safe ATP dependent DNase which was applied to digest the single strand DNA in rcDNA and ssDNA,then was quantitated by the primers spanning across the nick and SYBR Green Ⅰ dye.The specifity of PCR production was confirmed by the sequence analysis and rcDNA comparison.The significance of the difference of HBV cccDNA level between HBeAg(+) and HBeAg(-) group was analyzed by two group t test.Results The agarose gelelectrophoresis showed the molecular weight of the PCR production was about 350bp.The coincidence rate of PCR production and goal fragement was nearly 99% by sequence analysis.The result of PCR detection of rcDNA group was negative.The positive rate of HBV cccDNA of liver biopsies of HBeAg (+) patients detected by this method was 100%,the level of HBV cccDNA in the liver biopsies of HBeAg (+) patients was higher than HBeAb(+) patients.Conclusions The specificity of the method is proved by agarose electrophoresis,gene sequencing of the PCR product and rcDNA comparison.The quantitative method that use SYBR Green Ⅰ dye and ?-Globin as internal control is more specific,sensitive and economical,and more suitable for clinical purpose.

Objective To analyze the epidemiological characteristics of Mycobacterium tuberculosis by enterbaeterial repetitive intergenic consensus-polymerase chain reaction(ERIC-PCR)DNA fingerprint. Methods Mycobacterium tuberculosis positive sputum samples between September 2003 to May 2006 were collected and cultured.Chromosomal DNA were extracted and ERIC-PCR DNA fingerprinting was analyzed by software,such as RAPD PHYLIP and Treeview.Results A total of 42 different fingerprints were detected.Phylogenetic analysis showed that they could be classified into three clusters,the clustering rate was 72.6%.The characteristics of ERIC-PCR fingerprint patterns were related to age,drug resistance,and type of resistance.Conclusions ERIC-PCR DNA fingerprinting technique used in this study is good for epidemiological studies with its strong discrimination,simplicity and rapidness.A high level of recent transmission is found in our city.