Objective To develop an approach for a rapid extraction of genomic DNA fragments from serum.Methods Serum specimen from human was processed using high temperature denaturation and was centrifuged, then using various primer sets to amplify the DNA fragment extracted, so as to verify the efficiency of this DNA extraction method.Results The templates preparing from this DNA extraction method were able to guarantee the amplification of fragments shorter than 400 base pairs.Conclusion This method is simple, rapid and inexpensive.These genomic DNA fragment extracted can be utilized to amplify definite DNA template for polymerase chain reaction.

OBJECTIVE: To research the content of alkaloids and polysaccharides in different varieties of Aconitum carmichaeli. METHODS: The contents of total alkaloids, diester diterpenoid alkaloids and polysaccharides were measured by acid dye colorimetry, HPLC and sulphuric acid-anthrone colorimetric method respectively. RESULTS: In different varieties, the differences of total alkaloids and diester diterpenoid alkaloids were significant. However, the difference of polysaccharides wasn’t significant. The content of total alkaloids and diester diterpenoid alkaloids in processed A. carmichaeli reduced significantly while the content of polysaccharides increased slightly. CONCLUSION: The results of study could provide a scientific basis for rational breeding of A. carmichaeli.

To study the drug-drug interaction of morinidazole and warfarin and its application, a sensitive and rapid liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for the determination of R-warfarin/S-warfarin in human plasma. In a random, two-period crossover study, 12 healthy volunteers received a single oral dose of 5 mg racemic warfarin in the absence and presence of morinidazole. Blood samples were collected according to a pre-designed time schedule. R-warfarin, S-warfarin and methyclothiazide were extracted with ethylether : methylenechloride (3 : 2), then separated on a Astec Chirobiotic V (150 mm x 4.6 mm ID, 5 microm) column using 5 mmol x L(-1) ammonium acetate (pH 4.0) - acetonitrile as mobile phase at a flow-rate of 1.5 mL x min(-1). The mobile phase was splitted and 0.5 mL x min(-1) was introduced into MS. A tandem mass spectrometer equipped with electrospray ionization source was used as detector and operated in the negative ion mode. Quantification was performed using multiple reaction monitoring (MRM). The resolution of warfarin enantiomers is 1.56. The linear calibration curves for R-warfarin and S-warfarin both were obtained in the concentration range of 5 - 1 000 ng x mL(-1). Intra- and inter-day relative standard deviation (RSD) for R-warfarin and S-warfarin over the entire concentration range across three validation runs was both less than 10%, and relative error (RE) ranged from -4.9% to 0.7%, separately. The method herein described is effective and convenient, and suitable for the study of metabolic interaction between morinidazole and warfarin. The results showed that coadministration of warfarin with morinidazole did not affect the pharmacokinetics of either R-warfarin or S-warfarin.

ObjectiveTo evaluate left ventricular systolic synchrony in patients with coronary artery disease (CAD) by speckle tracking imaging (STI) and real-time three-dimensional echocardiography (RT3DE) and investigate the correlation with ECG-gated myocardial perfusion single-photon emission computed tomography (GMPS).MethodsA total of thirteen patients with CAD diagnosed by coronary angiography underwent STI and RT-3DE examinations.The data was analysed off-line using Qlab 8.0 software.STI systolic synchrony indexes included the standard deviation of times to peak strain in radial and circumferial direction in 12 left ventricular segments (Trs12-SD and Tcs12-SD),the standard deviation of times to peak longitudinal strain in 16 left ventricular segments (Tls16-SD).RT-3DE systolic synchrony indexes included the standard deviation of times to the minimum systolic volume in 16 and 12 left ventricular segments (Tmsv16-SD and Tmsv12-SD).GMPS was performed within one week before or after echocardiography.Phase analysis was performed offline using Emory Cardiac Toolbox software.Peak phase,phase SD,bandwidth,skewness and kurtosis were calculated.Results Trs12-SD derived from STI had a positive correlation with phase SD and bandwidth ( r =0.800,P ＜0.05 ; r =0.607,P ＜0.05).Tmsv16-SD derived from RT-3DE had a better positive correlation with phase SD and bandwidth ( r =0.847,P =0.001 ; r =0.890,P ＜0.001).ConclusionsTmsv16-SD derived from RT-3DE had a better correlation with GMPS parameters than STI parameters.RT-3DE assessment of left ventricular systolic parameters is expected to become the ideal synchronization indicator.

Objective To investigate therapeutic effects of Salvia miltiorrhiza on hepatic fibrosis of rats induced by carbon tewachloridean. Methods Rat models of hepatic fibrosis were founded by intraperitoneal injection of carbon tetrachloride. Salvia miltiorrhiza were given to these rats. Normal group and control group were set for comparison at the same time. Serum levels of ALT, AST, HA, LN, and PC-Ⅲ were detected; HE and Masson staining were conducted in hepatic tissues to observe pathological variations. Results Salvia miltiorrhiza could decrease serum levels of ALT, AST, HA, LN,PC-Ⅲ obviously (P ＜0.01), compared with the control group; Salvia miltiorrhiza could obviously improve pathological variations compared with the control group. Conclusion Salvia miltiorrhiza has therapeutic effect on hepatic fibrosis of rats

Objective To explore the changes of the hippocampal formation and the myelinated nerve fibers in the hippocampal formation of aged female Long-Evans rat. Methods The hippocampal formation and the myelinated nerve fibers in the hippocampal formation of 5 young (6-month old), 5 middle-aged (18-month old) and 6 aged (28-month old) female Long-Evans rats were quantitatively investigated with the stereological techniques and transmission electronic microscope technique. Results There were no significant changes in the volume of hippocampal formation, the volume density, the total volume, the length density and the mean diameter of the myelinated nerve fibers in the hippocampal formation among young, middle-aged and aged rats. When compared to that of young rats, the total length of the myelinated nerve fibers in the hippocampal formation of middle-aged rats was significantly increased by 63.6%. When compared to that of middle-aged rats, the total length of the myelinated nerve fibers in the hippocampal formation of aged rats was significantly decreased by 47.5%. When compared to that of young rats, the total length of the myelinated nerve fibers in the hippocampal formation of aged rats was non-significantly decreased by 13.8%. Conclusion Although the reason why the total length of the myelinated nerve fibers in the hippocampal formation of middle-aged rats was longer than that of young rats needs further investigations, the present results together with our previous findings in white matter and cortex further suggest that there are age-related changes of the myelinated nerve fibers in the normal aged brains.

Objective To investigate the relationship between amnestic mild cognitive impairment and functional genes associated with hyperphosphorylated tau protein.Methods One hundred and sixteen amnestic mild cognitive impairment (aMCI) patients and 93 normal controls were recruited for the study.Multi-dimension neuropsychologic tests were used to assess the cognitive function extensively.MassARRAY and iPlex systems were used to measure candidate SNP polymorphisms,analyze genotypic,allelic or haplotypic distributions and their interaction with ApoE ε4 and the correlation with the cognitive function in the subjects.Results ( 1 ) The scores of neuropsychologic tests in memory domain ( Auditory Verbal Learning Test (AVLT)-first immediate recall,AVLT-second immediate recall,AVLT-second immediate recall,AVLT-5 minute delayed recall,AVLT-20 minute delayed recall,AVLT-recognition,Rey-Osterrich Comolex Test-delay) in aMCI patients ( 3.0 ( 0-7.0 ),5.0 ( 1.0-10.0),6.0 ( 1.0-11.0 ),4.0 (0-11.0),3.0(0-10.0),20.0(8.0-24.0),11.2 ±8.3) were significantly lower than those in the normal controls(4.0(0-9.0),7.0(2.0-11.0),9.0(3.0-12.0),8.0(0-12.0),8.0(0-12.0),22.0 (10.0-24.0),16.1±8.0) (Z=-3.592,-6.802,-6.408,-8.173,-8.533,-5.647 andt=4.216 respectively,all P ＜0.01 ) ; (2) Genotypic distributions of rs242562 GG in aMCI (7.826％ ) were significantly lower than those in normal controls (20.65％,OR =0.3525,95％ CI 0.1411-0.8807,P =0.024 98),however there were no differences in the genotypic,allelic or haplotypic distributions between aMCI patients and controls of glycogen synthase kinase-3β,cyclin dependent protein kinase-5,calcium and calmodulin-dependent protein kinase-Ⅱ,cell division cycle 2,dual-specificity tyrosine-phosphorylation regulated kinase 1A and low density lipoprotein receptor-related protein 6; (3) MAPT/STH rs242562 genotype was correlated with AVLT-immediate recall,AVLT-delayed recall,Rey-Osterrieth Complex Test,Rey-Osterrieth Complex Test-delayed recall and Clock Drawing Test (H =9.763,12.258,10.508,9.624,10.767,F =3.700,3.123 and H =6.591 respectively,all P ＜ 0.05 ) ; (4) There were no differences in the distributions of MAPT/STH rs242562 GG genotype and ApoE ε4 haplotype between aMCI patients and normal controls.Conclusions MAPT/STH rs242562 GG genotype decreases the genetic risk of aMCI,which might have important role in memory function in aMCI.The interaction between rs242562 GG and ApoE ε4 doesn' t affect the susceptibility to aMCI.

The purpose of this study was to evaluate the effect of adenosine A2A receptor antagonist ZM241385 on amygdala-kindled seizures and its roles in epileptogenesis. Electrodes were implanted into the right amygdala of male adult Wistar rats. Kindling was accomplished by using stimulus strength of 500 μA applied daily to the amygdala until 10 consecutive stage 5 seizues were induced. Then effect of ZM241385 was studied in fully kindled rats after intracerebroventricular administration of the drug. In addition, the effect on kindling progression was evaluated through ZM241385 injection before daily stimulation. In all experiments, behavioral changes in the rats in response to ZM241385 were monitored closely. The results showed that, in fully amygdala-kindled rats, ZM241385 (0.001-0.1 nmol/L) decreased afterdischage duration (ADD), motor seizure duration (MSD), stage 5 duration (S5D) and seizure duration (SD), but only the effect on ADD was dose-dependent. The doses of 0.001-0.1 nmol/L had no influence on stage 4 latency (S4L) and seizure stage (SS). The dosages of 0.0001 and 1 nmol/L of ZM241385 did not exert any effect on all seizure parameters. In contrast to the results in fully amygdala-kindled rats, ZM241385 (0.001-0.1 nmol/L) had minimal or no effects on the progression of amygdala-kindled seizures. We are led to the conclusion that although ZM241385 had no influence on the progression of amygdala-kindled seizures, it had potent anti-convulsant profile and little adverse effects at the dosage of 0.001-0.1 nmol/L, suggesting that the agent is effective against the amygdala-kindled seizures.

AIM:To demonstrate the relationship between hormones in follicular fluid and the expression of LH receptor in granulosa cells(GC) in anovulatory women with polycystic ovary syndrome(PCOS).METHODS: Follicles were obtained from 12 women with PCOS and 15 women with normal menstrual period through surgery at time between day 7 and day 10 of menstrual cycle.The accumulations of estrogen(E2),progesterone(P),luteinizing hormone(LH),follicle stimulating hormone(FSH) and insulin in follicular fluid were determined by a automatism chemiluminescent microparticle immunoassay(CMIA) for the quantitative determination.The accumulation of androstenediol(A) was determined by ELISA.The amounts of the mRNA expressions of LH receptors from GC and theca cells(TC) respectively were measured by RT-PCR using ?-actin as intra-control simultaneously.RESULTS: The levels of LH [(3.8?2.1 vs 1.7?0.8)IU/L,P

Objective To construct the stable stathmin-overexpression SMMC-7721 hepatocellular carcinoma cells and to explore the effect of stathmin-overexpression on the cell proliferation and metastasis in SMMC-7721 cells .Methods By using liposome , Flag-pcDNA3 .1 and Flag-pcDNA3 .1-stathmin plasmid were transfected into SMMC-7721 cells respectively ,the stable Flag-pcD-NA3 .1 expression cells(control group) and the stable stathmin-overexpression cells(experimental group) were established after an-tibiotic resistant gene screening ,and the cell lines were identified by Western Blot .Subsequently ,the cell proliferation was detected by cell count kit(CCK-8) and the soft agar assay ,the apoptosis and cell cycle were determined by the flow cytometry (FCM ) ,and the cell motility and invasion were analyzed by the Transwell assay in vitro .Results The stathmin protein expression of the experi-mental group was significantly increased compared with the control group (0 .76 ± 0 .12 vs .0 .16 ± 0 .05 ,P<0 .05) ,which indicated that the stathmin-overexpression human SMMC-7721 hepatocellular carcinoma cell line was successfully constructed .CCK-8 and the soft agar assay showed that the cell proliferation of the experimental group was higher than that of the control group (0 .29 ± 0 .03 vs .0 .60 ± 0 .05 ,P< 0 .01);additionally ,the apoptotic ratio of the experimental group was decreased compared with the control group[(11 .57 ± 1 .09)% vs .(5 .80 ± 0 .33)% ,P<0 .05] ,the cell cycle was arrested in the stage G2/M ;the Transwell experiment results verified that the cell motility and the invasive ability of the experimental group were obviously reinforced compared with the control group[transmenbrane cell numbers in migrant assay :(54 .03 ± 7 .21) vs .(130 .45 ± 14 .13);transmenbrane cell numbers in invasive assay :(17 .75 ± 2 .52) vs .(57 .76 ± 8 .50) respectively] ,the differences had statistical significance(P<0 .01) .Conclusion The overexpression of stathmin promotes the cell proliferation and the invasive ability in SMMC-7721 hepatocellular carcinoma cells .