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 investigate the shape and function of right ventricles in patients received heart transplantation(HT). Methods Sixty healthy volunteers(control group) and 31 HT patients(HT group) were enrolled, and the HT group was further divided into the nonrejected(HTn) group and the rejected(HTp) group based on the endomyocardial biopsy(EMB) results. All the participants received routine echocardiography. Single beat real-time three-dimensional echocardiography (sRT-3DE) was performed in all to evaluate parameters concerning modality and systolic function of participants' right ventricles. Results 1) Right ventricular stroke volume (RVSV) and right ventricular ejection fraction (RVEF) were significantly different among the groups[RVSV: control group vs HTn group vs HTp group was (56.18 ± 23.72)ml vs (36.08 ± 10.94)ml vs (26.22 ± 9.84)ml, P ＜0.0001; RVEF:control group vs HTn group vs HTp group was (62.09± 7.18)% vs (51.04 ± 7.58) vs (35.86 ± 9.86)%, P ＜0.00001＝.2＝ When taking the influence of rejection into consideration,RVEF was proved to be a stable and sensitive indicator. Conclusions sRT-3DE can quickly assess shape and systolic function of right ventricle.RVEF was the most stableand sensitive among all the RV-related indicators and is a promising indicator in the clinic follow-up of HT patients.

Objective To examine supine fulcrum pressurized radiograph to predict spinal flexibility in adolescent idiopathic scoliosis (AIS). Methods 1. Empirical study: put two points (A and B) on one side of the plastic stick and one point on the other side ( C), pressurize on the plastic stick when point C is in the middle of A and B, and then measure the projection shifting of point A. Repeat the pressure test when the distance between A and C doubled. Analyze the effect of increasing distance on diminishing pressure. 2.Clinical study :45 patients ( 16 males and 29 females) underwent standing anteroposterior radiograph ,fulcrum lateral flexion radiograph, supine lateral flexion radiograph. The Cobb's angles were measured and the flexibility ratio was determined on preoperative fulcrum radiograph. Results (1) If the length of A and C was 25 cm, pressurize on point C, when pressure quantitative scale number located 1 scale and 2 scale of the rule, projection shifting of point A were 5 cm and 10 cm. If the length of A and C was 50 cm, pressurize on point C, when pressure quantitative scale number located 1 scale and 2 scale of the rule, projection shifting of point A were 8 cm and 15 cm. The longer the distance, the lesser the pressure force of plastics stick. (2)The Cobb angles and curve flexibility provided by fulcrum lateral flexion radiograph had significant difference from that provided by supine lateral flexion radiograph in male group and female group. Cobb angles of male group was 28. 6°± 4. 1 °, 16. 7°± 4. 6° respectively (t= 7. 438, P ＜ 0. 01 )and curve flexibility was 47. 6% ±8. 1% ,69. 4% ± 8. 5% respectively( t = 7. 438, P ＜ 0. 01 ). Cobb angles of female group was 24.5°± 2. 7°,12. 6°±2. 4° respectively(t = 17. 540,P ＜0. 01 )and curve flexibility was 53.4% ±5.2% ,76. 0% ±4. 7% respectively(t = 17. 54, P ＜ 0. 01 ). The flexibility of spinal main curve of female patients was obviously higher than male patients. Conclusions Supine fulcrum pressurized technique can make up for the insufficiency of supine lateral flexion technique, and provide a convenient, safe, precise and useful method for evaluation of spinal flexibility in AIS.

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 .

Objective To choose zebrafish as the experimental animal model for studying the spatiotemporal expression rule of rap1 gen in zebrafish embryo early development process .Methods The Rap1 gene fragment was cloned from the zebrafish emby‐oscDNA ,then the Rap1 gene fragment and pCS2+ plasmid were performed the in vitro connetion and recombination was extracted , the combinant plasmid was correct after the double enzyme digestion ,colony PCR and sequencing identification .T3 RNA polymer‐ase in vitro transcription system was used to obtain the digoxin (DIG)‐labeled anti‐sense mRNA probe of Rap1 gene .The whole mount in situ hybridization method was adopted to detect the Rap1 expression in zebrafish embryo early development process . Results The positive hybridization signal of Rap1 gene was detected at the cell division junction region of 0 .75 hpf ,animal pole of 3 .70 hpf and 6 .00 hpf ,and notochord of 12 .00 -72 .00 hpf .Conclusion Rap1 gene might be involved in the early development process of notochord nervous system in zebrafish .

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