A rapid, sensitive and convenient LC-MS/MS method was developed for the determination of γ-aminobutyric acid (GABA) in human plasma. d2-γ-Aminobutyric acid (d2-GABA) was synthesized as internal standard (IS). After extraction from human plasma by protein precipitation with acetonitrile, all analytes were separated on a Luna HILIC column (100 mm x 3.0 mm, 3 μm) using an isocratic mobile phase of water: acetonitrile: formic acid (20 : 80 : 0.12) with a flow rate of 0.5 mL x min(-1). Acquisition of mass spectrometric data was performed in multiple reaction monitoring mode (MRM) in positive electrospray ionization using the transitions of m/z 104 --> 69 for GABA and m/z 106 --> 71 for d2-GABA. The method was linear in the concentration range of 5.00 to 1 000 ng x mL(-1). The intra- and inter-day precisions were within 9.9%, and accuracy ranged from 99.1% to 104%, within the acceptable limit across all concentrations. The method was successfully applied to a pharmacokinetic study of GABA tablets in healthy Chinese volunteers.
Chromatography, Liquid ; Humans ; Tandem Mass Spectrometry ; gamma-Aminobutyric Acid ; blood

Objective of this study was to investigate the transcriptional regulation of BHLHB2 gene by the PML-RARα fusion protein in APL cells and reveal the pathogenesis of APL. RT-PCR was performed to detect the expression change of BHLHB2 before and after the induction of PML-RARα in PR9 cells, and its expression level after the treatment of ATRA in PR9 and APL patient derived NB4 cells. Chromatin immunoprecipitation (ChIP)-based PCR was used to analyze whether the BHLHB2 promoter could be bound by PML-RARα in vivo. A large-scale gene expression profile dataset was used to observe the expression pattern of BHLHB2 in AML. The results showed that the expression level of BHLHB2 was significantly reduced with the induction of PML-RARα and ATRA could reverse this inhibition in both PR9 and NB4 cells and increase the expression of BHLHB2. However, the expression of BHLHB2 could not be induced by ATRA in U937 cells which do not express PML-RARα. Mechanism study revealed that PML-RARα could bound to the promoter of BHLHB2 in vivo to regulate the the expression of BHLHB2. It was found that the expression of BHLHB2 was relatively lower in APL as compared with other subtypes of AML and normal bone marrow cells. It is concluded that BHLHB2 is the target of PML-RARα, and the expression of BHLHB2 is inhibited by PML-RARα through binding to its promoter in APL.
Basic Helix-Loop-Helix Transcription Factors ; genetics ; Gene Expression Regulation, Leukemic ; Homeodomain Proteins ; genetics ; Humans ; Leukemia, Promyelocytic, Acute ; genetics ; pathology ; Oncogene Proteins, Fusion ; genetics ; Promoter Regions, Genetic ; Transcription Factors ; genetics ; Tumor Cells, Cultured ; U937 Cells

We investigated the development of an injectable, biodegradable hydrogel composite of poly(trimethylene carbonate)-F127-poly(trimethylene carbonate)(PTMC11-F127-PTMC11 )loaded with bone morphogenetic protein-2 (BMP-2) derived peptide P24 for ectopic bone formation in vivo and evaluated its release kinetics in vitro. Then we evaluated P24 peptide release kinetics from different concentration of PTMC11-F127-PTMC11 hydrogel in vitro using bicinchoninic acid (BCA)assay. P24/ PTMC11-F127-PTMC11 hydrogel was implanted into each rat's erector muscle of spine and ectopic bone formation of the implanted gel in vivo was detected by hematoxylin and eosin stain (HE). PTMC11-F127-PTMC11 hydrogel with concentration more than 20 percent showed sustained slow release for one month after the initial burst release. Bone trabeculae surround the P24/ PTMC11-F127-PTMC11 hydrogel was shown at the end of six weeks by hematoxylin and eosin stain. These results indicated that encapsulated bone morphogenetic protein (BMP-2) derived peptide P24 remained viable in vivo, thus suggesting the potential of PTMC11-F127-PT- MC11 composite hydrogels as part of a novel strategy for localized delivery of bioactive molecules.
Animals ; Biocompatible Materials ; chemistry ; Bone Morphogenetic Proteins ; pharmacology ; Bone and Bones ; drug effects ; Dioxanes ; chemistry ; Drug Delivery Systems ; Hydrogels ; chemistry ; Osteogenesis ; drug effects ; Peptides ; Prostheses and Implants ; Rats

This is to report the screening, extracting and validating antitumor components and compounds from Stellera chamaejasme L. under the case of discrete distribution of active data. In this work, different components from Stellera chamaejasme L. were collected by HPD macroporous resin and polyamide resin column, and their antitumor activity on A549 were tested by MTT assay. Activity results indicate that activity of components at 30-39 min is more potent than that of Stellera chamaejasme L. extract, and the activity of components at 33.97 min is equivalent to positive drug, cis-platinum at 100 microg x mL(-1), but with totally different mode of action. Under the case of discrete activity, the weight analysis is capable of screening active components and compounds from natural products.
Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Drug Screening Assays, Antitumor ; Humans ; Thymelaeaceae ; chemistry

This article studies the thermostability and the crystal structure of a new anticancer drug dasatinib. The thermostability of dasatinib was analyzed using the differential scanning calorimeter (DSC) and thermo gravimetric analyzer (TGA), and the structural characteristics of polymorphism and crystalline transformation was determined using the X-ray powder diffractometry (XRD) with in-situ high temperature accessories. The results showed that dasatinib has at least two different crystal forms. The form-I has one crystalline water and form-II one and half, and in a heating-up processing both of them would change their crystal structures. After losing their crystalline water, both would change into the same crystalline form with no crystalline water. Their melting points were almost the same: form-I was 285.68 degrees C and form-II was 285.50 degrees C. The results of the study method would provide a comprehensive reference for the quality evaluation of dasatinib.
Antineoplastic Agents ; chemistry ; Calorimetry, Differential Scanning ; Crystallization ; Dasatinib ; Molecular Structure ; Protein Kinase Inhibitors ; chemistry ; Pyrimidines ; chemistry ; Temperature ; Thermogravimetry ; Thiazoles ; chemistry ; X-Ray Diffraction

OBJECTIVETo bioinformatically predict and analyze target genes of miRNA-126(*), with the aim of providing certain basis for related research about target genes and regulatory mechanism in the future.

METHODSThe miRNA chip technology was applied to measure expression levels of miRNA-126(*) in 3 time points (embryo 16, 19 and 21 days) of fetal lung development. Then the target genes of miRNA-126(*) were screened through miRGen2.0 database. Subsequent bioinformatic analysis of these target genes was performed by Gene Ontology analysis and Kyoto Encyclopedia of Genes and Genomes Pathway analysis (KEGG Pathway analysis).

RESULTSmiRNA-126(*) manifested continuously upregulated expression with the lung development (from embryo 16 to 21 days). There were 422 predicted target genes in total, and the gene set mainly located in glucuronosyltransferase activity, transferase activity (GO molecular function), multicellular organismal development, developmental process (GO biology process) and intracellular part (GO cellular component). The KEGG Pathway analysis demonstrated that the gene set mostly located in RNA degradation (signal transduction pathway) and prion diseases (disease pathway).

CONCLUSIONSThe results suggest that miRNA-126(*) plays a certain role in fetal lung development and provide a basis for lung development research in the future.

Animals ; Computational Biology ; Female ; Glucuronosyltransferase ; metabolism ; Lung ; embryology ; Male ; MicroRNAs ; physiology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; physiology

OBJECTIVETo discuss the radical treatment of cervical giant cystic lymphangioma in Children and cosmetic result.

METHODSTwenty-five children with cervical giant cystic lymphangioma were retrospectively analyzed. The diameter of all the tumors was more than 10 cm. 24 cases underwent resection. The complication, therapeutic effect and cosmetic result were recorded.

RESULTSThe tumors were all removed radically in all the cases. The patients were followed up for 1-5 years with no recurrence. Cosmetic result was satisfactory in 22 cases. Secondary operation was performed in 2 cases with satisfactory result. Complications included 5 cases of lymph leakage, 2 cases of poor wound healing, 1 case of infection and 2 cases of tongue edema.

CONCLUSIONSThe cervical giant cystic lymphangioma in children can be resected radically with satisfactory result.

Child, Preschool ; Female ; Head and Neck Neoplasms ; surgery ; Humans ; Infant ; Lymphangioma, Cystic ; surgery ; Male ; Neck ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo investigate the association between the level of serum testosterone and atherosclerosis in middle-aged and elderly men.

METHODSWe conducted a population-based study of 413 males aged 40-75 years in a community in Guangzhou. We obtained the sociodemographic characteristics, clinical data, physical measurements, and laboratory results of sex hormones, blood glucose, and blood lipid of the subjects. We also measured the carotid intima media thickness (CIMT) by color Doppler ultrasonography.

RESULTSThe subjects were divided into a carotid atherosclerosis (CAS) group (CIMT ≥ 0.9 mm) and a non-CAS group (CIMT < 0.9 mm). The medians of free testosterone (FT) were 57.41 and 59.72 pmol/L in the CAS and non-CAS groups, respectively (P = 0.005), and no significant difference was found between the two groups in total testosterone (TT). The levels of serum FT and TT were negatively correlated to CIMT, with Spearman's rank correlation coefficients of -0.126 (P = 0.011) and -0.188 (P < 0.001), respectively. The incidence rates of CAS were 23.30, 13.46, 17.48, and 7.77% in the Q1, Q2, Q3, and Q4 groups, respectively according to the quartile of FT (P for trend = 0.008) and 17.48, 18.27, 16.50, and 9.71% respectively according to the quartile of TT (P for trend = 0.116). Based on the quartile of FT and after adjustment for age, waist circumference, systolic blood pressure, and HbAlc, the risk of CAS was significantly increased in the Q1 group as compared with Q4 (OR = 2.491, 95% CI 1.01-6.149), but no statistically significant differences were observed according to the quartile of TT.

CONCLUSIONA low serum FT level may be a risk factor of atherosclerosis in Chinese men aged 40 years or older.

Adult ; Age Factors ; Aged ; Blood Glucose ; analysis ; Blood Pressure ; Carotid Artery Diseases ; blood ; epidemiology ; etiology ; Carotid Intima-Media Thickness ; Gonadal Steroid Hormones ; blood ; Humans ; Incidence ; Lipids ; blood ; Male ; Middle Aged ; Risk Factors ; Statistics, Nonparametric ; Testosterone ; blood

OBJECTIVETo establish corresponding set pair analysis mathematic model using the clinical symptoms of blood-heat type psoriasis vulgaris (BHTPV), thus guiding the clinical accuracy of clinical syndrome typing.

METHODSRecruited were 202 BHTPV with complete data after they were treated by yang subduing blood cooling method. Their clinical symptoms and signs were systematically collected. Using set pair analysis method, the therapeutic results of cured and markedly effective were recruited as the same portion, of effective as the different portion, and of ineffective as the contrary portion. The U value of each syndrome factor was calculated according to the formula. The correlation factor of syndrome typing of BHTPV was screened. The syndrome typing model of BHTPV was established according to the correlation factor.

RESULTSThe main factors of BHTPV included the scale integral > 2.04, the erythema integral > 2.34, age > 50 years old, the area integral > 2.07, dry mouth, slippery pulse, yellowish fur, soggy pulse, dry and hard stool. The secondary factors included less sweat, insomnia, frequent pulse, any infiltration, erythra of any area, red tongue, depression, the disease course ranging 1-360 months, age ranging 16-50 years old, string-tight pulse, thin fur, the area integral ranging 0-2.07, white fur, purplish tongue, the scale integral ranging 0+ -2.04, and feeble pulse. The third factors included the erythema integral ranging 0-2.34 and pale red tongue. The set pair analytical model of BHT-PV was as follows: U = sigma An/N + sigma B(m)i/M + sigma C(p)j/P.

CONCLUSIONSU blood-heat syndrome > or = 0.75 could be judged as blood-heat syndrome. Satisfactory efficacy could be achieved by blood cooling method. For patients with U blood-heat syndrome < 0.75, no satisfactory efficacy could be achieved by blood cooling method alone since they were accompanied with other syndrome types.

Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Psoriasis ; diagnosis ; Young Adult

OBJECTIVETo compare the transcription difference of the mannitol PTS genes between epidemic and non-epidemic strains of Vibrio cholerae El Tor in mannitol ferment tests.

METHODSGrowth curves of 10 epidemic strains (slow-ferment) and 10 non-epidemic strains (rapid-ferment) of Vibrio cholerae were detected in the process of fermentation test, and the transcriptional level of mannitol PTS operon of these strains were determined with quantitative reverse-transcriptional PCR.

RESULTSAfter 4 hours of test, the non-epidemic strains became positive and the average growth density of the non-epidemic strains was higher than that of the epidemic strains; however, some were still lower than the epidemic strains. In contrast, at the eighth hour of test, when epidemic strains got positive, they showed higher average growth density. Compared to the epidemic strains, the transcription of mannitol PTS genes of the non-epidemic strains were much more active at the 1st and 2nd hour and were lower at the 4th and 8th hour.

CONCLUSIONSThe difference of mannitol PTS operon transcription level should be an important feature to identify the epidemic and non-epidemic strains of Vibrio cholerae, which directly influences the mannitol fermentation rate during the test. The growth rate is not a key factor that affect such difference.

Bacterial Proteins ; genetics ; Gene Expression Regulation, Bacterial ; Mannitol ; metabolism ; Operon ; Phosphoenolpyruvate ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sugar Alcohol Dehydrogenases ; metabolism ; Transcription, Genetic ; Vibrio cholerae ; classification ; genetics ; metabolism