Adult ; Burnout, Professional ; Female ; Humans ; Life Change Events ; Middle Aged ; Occupations ; Professional Competence ; Work ; psychology ; Young Adult

A mutant strain RZ13 with high lipase productivity was obtained through treating the Rhizopus sp. RXF12 by UV and microwave,and its activity was 2.62-fold of the original one. The high lipase productivity of the mutant strain could be inherited after repetitious subcultures.The optimal culture conditions of the RZ13 for producing lipase were studied.The lipase with highest activity (67.32U/ml) was obtained on the conditions of 25℃,pH 8.0,5 %(v/v) single spore suspension (1?107spore/ml) and 120 r/min for 84h,and stable under 40℃ and pH 8.5. In addition,the lipase was immobilized by adsorption onto Mg-Al hydrotalcite at 25℃ for 4 h by screening carriers and optimizing the immobilization. The results showed that the optimal reaction temperature and pH were 35~55℃ and 7.5~9.0,respectively,much wider than the free lipase.

OBJECTIVETo study the stereoselectivity of R-(+) and S-(-)-propranolol glucuronidation and metabolic interaction between R(+)- and S-(-)-propranolol.

METHODSA RP-HPLC analytical method was developed for determination of R-(+)-and S-(-)-propranolol glucuronide (PG) incubated with rat hepatic microsome induced with phenobarbital (PB). The method was applied to investigate the stereoselectivity metabolism of racemic propranolol glucuronidation in vitro.

RESULTIn control and PB group, the concentration of R-(+)-PG produced at different substrates was higher than that of S-(-)-PG. Compared with the control, the V(max) and Cl(int) for R(+)-and S-(-)-propranolol increased significantly the K(m) for R(+)-propranolol was elevated, while that for S-(-) propranolol was decreased.

CONCLUSIONThere is a stereoselectivity in glucuronidation of propranolol in rat hepatic microsome induced with PB and R-(+)-propranolol is preferred. Metabolic interaction between R-(+)-and S-(-)-propranolol exists with a concentration-dependent mode.

Animals ; Enzyme Induction ; Microsomes, Liver ; enzymology ; Phenobarbital ; pharmacology ; Propranolol ; analogs & derivatives ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stereoisomerism

Objective:To investigate the expression of H19 long non-coding RNA (IncRNA) and zinc finger E-box-binding protein i (ZEB1) in the trophoblast of women with spontaneous abortion.Methods:A total of 20 women underwent miscarriage were enrolled as a miscarriage group,while 20 women underwent artificial abortion were enrolled as a control group.Reverse transcriptional fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of iH19 lncRNA and miR-200,and Western blot was used to detect the expression of ZEB1.Results:Compared to the control group,a tendency of decreased H19 was detected in the miscarriage group,with no significant difference (1.28±0.08 vs 1.66±0.21,P=0.091).There was no significant difference in miR-200 between the 2 groups (0.75±0.35 vs 0.58±0.33,P=0.0.533).The expression of ZEB 1 was significantly decreased in the miscarriage group compared to that in the control group (0.22±0.03 vs 0.41±0.04,P=0.0003).The expression of i19 lncRNA and ZEB1 was positively correlated (r=0.529,P=0.0005),and the correlation between H19 lncRNA and miR-200 had no statistical significance (r=0.0293,P=0.858).The correlation between miR-200 and ZEB1 also had no statistical significance (r=-0.132,P=0.416).Conclusion:Down-regulation of ZEB1 in the trophoblast might be related to miscarriage.H19 lncRNA may regulate the expression of ZEB1.

BACKGROUNDCandida albicans is the most frequently seen opportunistic human fungal pathogen. Terbinafine is an allylamine antifungal agent that has been proven to have high clinical efficacy in the therapy of fungal infections, the mechanism of action of terbinafine involves the specific inhibition of fungal squalene epoxidase, resulting in ergosterol deficiency and accumulation of intracellular squalene. We used cDNA microarray analysis technology to monitor global expression profile changes of Candida albicans genes in response to terbinafine treatment, and we anticipated a panoramic view of the responses of Candida albicans cells to the representatives of allylamine antifungal agents at the molecular level in an effort to identify drug class-specific and mechanism-independent changes in gene expression.

METHODSCandida albicans strain ATCC 90028 was exposed to either medium alone or terbinafine at a concentration equivalent to the 1/2 minimal inhibitory concentrations (MICs, 4 mg/L) for 90 minutes. RNA was isolated and gene expression profiles were compared to identify the changes in the gene expression profile using a cDNA microarray analysis. Differential expression of 10 select genes detected by cDNA microarray analysis was confirmed by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSA total of 222 genes were found to be responsive to terbinafine, including 121 up-regulated genes and 101 down-regulated genes. These included genes encoding membrane transport proteins belonging to the members of the ATP-binding cassette (ABC) or major facilitator superfamily (MFS; CDR1, AGP2, GAP6, PHO84, HOL3, FCY23, VCX1), genes involved in stress response and detoxification (CDR1, AGP2, HOL3), and gene involved in the ergosterol biosynthesis pathway (ERG12). The results of semi-quantitative RT-PCR were consistent with that of the cDNA microarray analysis.

CONCLUSIONSThe up-regulation of the gene encoding the multidrug resistance efflux pump CDR1 may contribute to the terbinafine resistance in Candida albicans. However, the precise roles of other affected genes remain unclear, further studies of these genes and their respective products that play roles in the context of antifungal resistance are warranted.

Antifungal Agents ; pharmacology ; Candida albicans ; drug effects ; genetics ; Ergosterol ; biosynthesis ; Fungal Proteins ; genetics ; Gene Expression Profiling ; Genome, Fungal ; Membrane Transport Proteins ; genetics ; Naphthalenes ; pharmacology ; Oligonucleotide Array Sequence Analysis

The present study was to investigate in vitro the rat cardiomyocyte injury with targeting of home-made perfluorocarbon lipid particles with avidin-biotin interaction. Neonatal rat cardiomyocytes were cultured in vitro and divided into two groups: TNF-alpha activated group and non-activated group. Those in the TNF-alpha activated group were exposed to 200 ng/ml TNF-alpha solution for 6 hours and then cardiomyocytes in both groups were pretargeted with biotinylated ICAM-1 monoclonal antibodies, and were exposed to streptavidin, and then to homemade green fluorescently-labeled biotinylated perfluorocarbon lipid particles. Cardiomyocytes nucleus stained with Hoechst. The results were detected with fluorescence microscope. As a result, in TNF-alpha activated group, around blue fluorescent cardiomyocytes nucleus, a great amount of green fluorescent particles were found, while there were few green fluorescent particles in non-TNF activated group. It has been shown that ICAM-1 is expressed in the surface of cardiomyocytes when they are stimulated by TNF-alpha. Perfluorocarbon lipid particles associated with ICAM-1 monoclonal antibodies can be targeted to injured cardiomyocytes by avidin-biotin interaction.
Animals ; Animals, Newborn ; Antibodies, Monoclonal ; metabolism ; Cells, Cultured ; Contrast Media ; Female ; Fluorocarbons ; immunology ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipids ; chemistry ; Male ; Microspheres ; Myocytes, Cardiac ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; pharmacology ; Ultrasonography

Objective　Coronary artery bypass grafting (CABG)i n 13 patients was analyzed. Methods　9 patients were performed C ABG with cardiopulmonary bypass, 4 patients undergone off-pump coronary artery bypass(OPCAB). Among the 4 patients, 3 undergone transmyocardial laser revascula rization concomitantly. 2 patients with single-vessel disease, 3 with double-v essel disease, 7 with triple-vessel disease and 1 with left main coronary arter y disease. The average bypass per patient was 2.3. Results　All patients survived, 11 patients were angina free, 2 were angina relief. C onclusion　CABG is a safe operation, OPCAB may reduce blood transfusion and complication, patients recover more quickly after OPCAB compared with those with CABG.

OBJECTIVETo explore the association between angiotensin-converting enzyme (ACE) and the polymorphisms of N5, N10-methylenetetrahydrofolic acid reductase (MTHFR) gene in patients with ischemic stroke (IS).

METHODSTotally 454 patients with IS (IS group) and 334 controls (control group) were recruited in our study. Their I/D polymorphisms of ACE gene and C677T polymorphisms of MTHFR gene were detected by PCR and denaturing high performance liquid chromatography.

RESULTSThe frequencies of DD, ID, II and CC, CT, TT genotype in IS group were 22.5%, 43.4%, 34.1%, and 51.8%, 40.5%, 7.7%, respectively, and were 17.4%, 45.5%, 37.1% and 56.9%, 38.3%, 4.8% in the control group, respectively. DD genotype was associated with large-artery atherosclerosis (LAA), and TT genotype and T allele were associated with LAA and cardioembolism. Synergistic effects were found between TT and DD/ID DD genotypes in the pathogenesis of ischemic stroke.

CONCLUSIONDD, TT genotype and T allele are risk factors of IS, and ACE gene and MTHFR gene have synergistic effects in the pathogenesis of IS.

Brain Ischemia ; complications ; genetics ; Genetic Predisposition to Disease ; Humans ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Polymorphism, Genetic ; Renin ; genetics ; Stroke ; etiology ; genetics

OBJECTIVETo establish a method for detecting the polymorphism of methylenetetrahydrofolate reductase gene (MTHFR).

METHODSThe MTHFR was amplified, and the amplified products were detected by denaturing high performance liquid chromatography (DHPLC), and the amplified MTHFR was confirmed by sequencing and restriction enzyme digesting.

RESULTSA total of 334 individuals of Han people in southern China were recruited in our study, and their polymorphisms of MTHFR were detected. The accurate rate of the DHPLC method, that was very sensitive with 100% detection rate available, was over 99%. The frequencies of CC, CT and TT genotypes were 56.9%, 38.3% and 4.8% individually, and the frequencies of T and C alleles were 23.95% and 76.05% individually.

CONCLUSIONThe DHPLC method can detect polymorphism of MTHFR rapidly, effectively and economically. And there is the existence of different MTHFR polymorphisms in area and race.

Adult ; Aged ; Aged, 80 and over ; Alleles ; China ; ethnology ; Chromatography, High Pressure Liquid ; methods ; DNA Mutational Analysis ; Female ; Humans ; Male ; Methylenetetrahydrofolate Dehydrogenase (NAD+) ; genetics ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Middle Aged ; Nucleic Acid Amplification Techniques ; Polymorphism, Genetic

AIMTo investigate the role of extracellular-signal regulated kinase (ERK) cascade on cerebral ischemia and ischemic preconditioning in hippocampal neuron.

METHODSMale gerbils were randomly divided into sham group (SH), ischemia/reperfusion group (I/ R), ischemia preconditioning group (IP), specific antagonist of ERK-PD98059 (PD), solvent control groups (VE group), PD98059 combined with IP group (PIP). Forebrain ischemia was induced by occlusion of bilateral common carotid arteries and confirmed by isoelectricity of EEG. Observations were carried out in each group 15 min, 2 h, 4 h, 6 h, 1 d, 3 d, 5 d and 7 d after ischemia. Open field test was used to examine the spontaneous motor activity, the survival and apoptotic neurons, Fos and NF-kappaB masculine neurons in hippocampal CA1 region were counted, the expression of HSP70 in hippocampal CA1 region and p-ERK in hippocampal CA3/DG regions were detected by SABC immunocytochemical technique.

RESULTSThe spontaneous motor activity, the number of apoptotic neurons and NF-kappaB masculine neurons at 1 d, 3 d, 5 d, 7 d in CA1 region were much less in IP group than in I/R group (P < 0.01). The number of Fos masculine neurons at 15 min, 2 h, 4 h, 6 h, 1 d in CA1 region were significant more in IP group than in I/R group (P < 0.01). The expressions of p-ERK and HSP70 were significantly higher in IP group than in I/R group. The number of Fos masculine neurons at each point were more and apoptotic neurons at 1 d, 3 d were less in PD group than in I/R group. Results of observation in PIP group were within IP group and I/R group.

CONCLUSIONActivation of ERK in CA3/DG regions were related to ischemic tolerance. Induction of the expression of Fos and HSP70, decreasing of the product of NF-kB which might be one of the molecule mechanisms playing an important role in neural protection of ischemic preconditioning.

Animals ; Brain Ischemia ; metabolism ; prevention & control ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Gerbillinae ; Hippocampus ; blood supply ; cytology ; Ischemic Preconditioning ; Male ; NF-kappa B ; metabolism ; Neurons ; metabolism ; Signal Transduction ; physiology