Objective To develop a sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of forsklin in rat plasma.Methods After extraction with methyl tert-butyl ether,chromatographic separation was performed on a C18 column with the mobile phase consisting of water ( 0.1％ formic acid)-acetonitrile in a gradient elution mode.A tandem mass spectrometer equipped with electrospray ionization (ESI) source was used as detector in the positive ion mode.Quantification was performed using multiple reaction monitoring (MRM) with the precursor product combination ions of m/z 411→375.3 and 285→193 for forsklin and diazepam.Results Good linearity was obtained in the 0.5-1000 ng/ml range for the analyte and the analytical method was validated in terms of specificity,precision,accuracy,recovery,stability and matrix effect.These assays gave RSD values always lower than 14.4％ and RE values between -3.5 ％ and 3.8％.In addition,the specificity,extraction recovery,stability and matrix effect were satisfactory.Conclusion Due to its high sensitivity,specificity and simplicity,the method could be used for pharmacokinetic studies of forsklin.

Objective To evaluate the effect of nicotine pretreatment on cardiac function following myocardial ischemia-reperfusion (I/R) in rats.Methods Sixty male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 3 groups (n =20 each):sham operation group (group S),group I/R and nicotine pretreatment group (group N).The rats were anesthetized with intraperitoneal 20％ urethane 1 g/kg,tracheostomized and mechanically ventilated.Myocardial ischemia was induced by occlusion of the left anterior descending branch of coronary artery for 30 min followed by 120 min of reperfusion.The left anterior descending branch of coronary artery was only exposed,but not occluded in group S.Nicotine 400 μg/kg was injected intravenously via the right jugular vein at 30 min before myocardial ischemia in group N.The equal volume of normal saline was injected instead in groups S and I/R.Before ischemia,at 30 min of ischemia and at 30 and 120 min of reperfusion,10 rats from each group were chosen for record of left ventricular systolic pressure (LVSP),left ventricular diastolic pressure (LVDP),± dp/dtmax,HR and mean arterial pressure (MAP).Blood samples were collected from the right carotid artery of the left 10 rats in each group at 60 min of reperfusion to measure plasma CK-MB activity and cTnI and TNF-α concentrations.Results Compared with group S,MAP and LVSP at T2,3 and HR,LVDP and ± dp/dtmax at T1-3 were significantly decreased,and the plasma CK-MB activity and cTnI and TNF-α concentrations were increased in group I/R,and LVDP at T1,2 and HR and ± dp/dtmax at T1-3 were significantly decreased,and the plasma CK-MB activity and cTnI and TNF-α concentrations were increased in group N (P ＜ 0.05).Compared with group I/R,MAP,HR,LVSP,LVDP and ± dp/dtmax were significantly increased at T3,and the plasma CK-MB activity and cTnI and TNF-α concentrations were decreased in group N (P ＜ 0.05).Conclusion Nicotine pretreatment can reduce myocardial I/R injury through activating cholinergic anti-inflammatory pathway,thus improving cardiac function in rats.

We have designed and carried out problem based learning (PBL) pedagogy since 2004. According to clinical eases, students learnt the pathophysiology of heart failure knowledge by themselves. Each group recommen-ded one student to make an oral presentation and wrote a review about heart failure. Preparing clinical cases and group discussions are very important. At the same time we should pay attention to the change in role of the teacher in PBL and cooperation with other disciplines.

Objective To assess the effects of health management on high-risk diabetic populations.Methods A total of 307 diabetic high-risk adults from 6 communities of Tianjin were recruited by using diabetes risk screening technology.Three-month intensive health management and nine-month follow-up were conducted in this participants.Paired t test for continuous variables and paired contingency table x2 test were used for data analysis.Results Energy intake (1989.8 vs.1766.4 kcal,t =6.84,P ＜0.05),effective exercises (120.4 vs.157.5 kcal,t =-5.00,P ＜ 0.05),body weight (73.0 vs.71.5 kg,t =6.92,P ＜0.05),systolic blood pressure (130.4 vs.124.6 mm Hg (1 mm Hg =0.133 kPa),t =8.36,P ＜0.05),diastolic blood pressure (81.8 vs.78.4 mm Hg,t =7.40,P ＜ 0.05),serum total cholesterol (5.21 vs.5.08 mmol/L,t =2.73,P ＜ 0.05),fasting plasma glucose (6.4 vs.5.8 mmol/L,t =16.37,P ＜ 0.05)and 2 h postprandial blood glucose (7.7 vs.6.9 mmol/L,t =9.67,P ＜ 0.05) were significantly improved after the intervention.Conclusions Community-based health management may provide an effective way to prevent and control the risk factors of diabetes.

Objective To assess the role of health education in outcomes of diabetes mellitus among high-risk populations.Methods The community physicians who participated this investigation received standardized training,and 307 community residents at high risk of developing diabetes obtained three-month intense health education and nine-month follow-up study.Paired t-test,and Analysis of Variance were used for data analysis.Results After systematic health education,professional level of community physicians was improved.Cognitive level of health knowledge was also significantly improved (5.5 vs 12.6,t=-28.511,P＜0.05).In addition,health knowledge of variant age (F=4.036,P＜0.05),education level (F=15.27,P＜0.05) and occupation (F=9.80,P＜0.05) subgroups was significantly increased.In comparison with baselines,the scores of each age subgroups (F=0.204,P＞0.05) showed no significant differences,although scores of different education level (F=4.71,P＜0.05) and occupation (F=4.87,P＜0.05) subgroups were significantly different.The risk factors of diabetes were effectively controlled.Conclusions Health education should be the key to health management of diabetes,which plays important roles in improving cognitive level of health knowledge among populations at high risk of developing diabetes and reducing the incidence of this condition.

Healthy Beagle dogs were administrated with batroxobin by intravenous infusion at high, medium and low doses. The study of pharmacodynamics and pharmacokinetics was intended to clarify the relevance of them and provided strong evidence for clinical use of batroxobin. The blood samples were collected after injection based on the time schedule and samples were tested by ELISA method to get the concentration of batroxobin. At the same time, changes of prothrombin time (PT), thrombin time (TT), activated partial thromboplastin time (APTT), fibrinogen (Fib) and D-dimmer were tested. The results showed that the concentration of D-D increased significantly after administration compared with that of before administration. The main pharmacokinetic parameters were as follows: t1/2 were (2.27 +/- 0.42) h, (10.65 +/- 2.19) h and (11.01 +/- 3.51) h; C(max) were (11.9 +/- 1.72) ng x mL(-1), (154.53 +/- 12.38) ng x mL(-1) and (172.14 +/- 47.33) ng x mL(-1); AUC(last) were (29.38 +/- 3.69) ng xh x mL(-1), (148.43 +/- 72.85) ng x h x mL(-1) and (599.22 +/- 359.61) ng x h x mL(-1). The elimination of batroxobin was found to be in accord with linear kinetics characteristics. The results of pharmacodynamics showed that D-dimmer level increased significantly after the administration of batroxobin, which was similar with the changes of batroxobin plasma concentration. Simultaneously, Fib concentrations in Beagle dog blood decreased significantly after the iv administration of batroxobin, while recovered to base level after 48 hours. PT, TT and APTT significantly became longer after administration, which returned to normal level after 48 hours. Especially, the D-dimmer levels and the batroxobin concentration in plasma after intravenous infusion of the drug were synchronized in Beagle dogs. Changes between PD/PK results had obvious correlation, and the D-dimmer levels in plasma can be one of the important monitoring indicators of batroxobin in thrombolytic medication.

Objective To study the effect of different approach on myocardial contractile force, angiogenesis and collagen renovation in acute myocardial infarct site by implantation of bone marrow mesenchymal stem cells (MSCs). Methods 32 Gui-zbou Xiang porcine were randomly divided into control group (group C), intra-coronary arterial injection group (group A) ,topical injection group (group T) and intra-coronary arterial + topical injection group( group A + T). 3 ml bone marrow was extracted. MSCs were cultured according to the methods of Wakitani's. After being co-cultured with 5-azacytidine for 24 hours, these cells were labelled with bromodeoxyuridine ( BrdU ). Autologus MSCs were implanted into the acute myocardial infarct site via the distal segment of the ligated left anterior descending artery (LAD) ,topical injection and both. Specimens and muscle strip were harvested in 3 and 6 weeks after MSCs implantation for microvessel count, P Ⅲ NP activity and contractile force measurement. Results The contractile force(48.6±5.9) %, (42.1±6.2 ) %, ( 56.9±5.1 ) %, microvessel count ( 19.6±4.3,17.1±4.0, 23.2±5.5 ) and collagen renovation (4.6±0.5 ) μg/L, ( 5.9±0.7 ) μg/L, ( 3.9±0.3 ) μg/L in the cell implant groups was better than that in the control group(37.9±5.4)% ,13.2±3.8,(8.7±0.8) μg/L,(P <0.01) ,espe-cially in group A + T. Conclusions Implantation of MSCs through intra-coronary arterial plus topical injection of MSCs maybe an ideal approach for infracted myocardium.

OBJECTIVE: To investigate the role of p27 in the inhibition of emodin on the mesangial cell (MC) proliferation induced by tumor necrosis factor-alpha(TNF-alpha). METHODS: p27 protein of MC was detected with western blotting analysis. The degree of MC proliferation was estimated through [(3)H] thymidine ([(3)H] TdR) incorporation. Different dosage of emodin (50 mg/L,100 mg/L) was added into MC stimulated by TNF-alpha. RESULTS: TNF-alpha (200 kU/L) decreased p27 level of MC cultured in serum-free DMEM for 24 hours and increased[(3)H] TdR incorporation. Emodin increased p27 level of MC stimulated by TNF-alpha and decreased [(3)H] TdR incorporation. The more the emodin was added, the greater the above-mentioned effect of emodin. CONCLUSION: The increment of p27 level maybe play an important role in the inhibition of emodin on MC proliferation induced by TNF-alpha.

Objective To investigate the relationship between Wnt5a gene and E-cadherin or vimentin gene in breast cancer cell line MCF-7. Methods RT-PCR was used to detect the mRNA expression of Wnt5a, E-cadherin and vimentin in breast cancer MCF-7 cells and the normal human mammary epithelial cell line MCF-10A, respectively, and their correlation was analyzed. Results The mRNA expression levels of Wnt5a and E-cadherin in cell line MCF-7 were significantly lower than those in cell line MCF-10A [(16.93± 2.97)%vs. (27.47±2.76) %, (12.97±1.35) % vs. (20.43±2.60) %, both P<0.05]. The mRNA expression level of vimentin in cell line MCF-7 was significantly higher than that in cell line MCF-10A [(16.53±0.85)%(6.33± 2.08) %, P<0.05 ]. In cell line MCF-7, the expression of Wnt5a was positively related to E-cadherin (г=0.997, P<0.05), but it was negatively related to vimentin (г=-0.998, P<0.05). Conclusions The expression of Wnt5a in human breast cancer cell line MCF-7 is significantly lower than that in cell line MCF-10A, which indicates that Wnt5a is a cancer suppressor gene in breast cancer. The expression of Wnt5a in cell line MCF-7 is positively related with E-cadherin, and it is negatively related with vimentin. Wnt5a may cause invasion and metastasis of breast cancer cell through the breast epithelial mesenchymal transitions.

Objective:To explore the clinical effectiveness of an automatic computer-aided diagnosis(CAD) method in benign and malignant breast masses discrimination.Methods:The ultrasound images of 539 patients from the Second Hospital of Harbin Medical University and the Second Hospital of Hebei Medical University between 2012 to 2019 were analyzed retrospectively. According to the fifth Breast Imaging Reporting and Data System (BI-RADS), four breast radiologists first sent the case into a BI-RADS category with the original ultrasound image. Then with the CAD result, radiologists gave a category again. Pathology results and clinical data were not available to the radiologists during the diagnosis process. The histological and follow-up results were used as the gold standard. The accuracy of CAD automatic classification, radiologists′ diagnosis before and after CAD application were compared using the ROC curves. The accuracy, sensitivity and specificity of the diagnosis were also calculated.Results:The classification algorithm has a good performance in benign and malignant breast masses discrimination.When the cutoff value was 0.495, the accuracy, sensitivity and specificity were 0.878, 0.868 and 0.886 respectively. When the cutoff value was 0.203, the sensitivity and specificity was 0.981 and 0.337 respectively. With the CAD method, the radiologists improved their diagnostic performance. The total area under the ROC curve for the four radiologists increased from 0.775 to 0.871( P<0.001). The total sensitivity increased from 0.786 to 0.842, and the specificity increased from 0.681 to 0.813. Conclusions:The automatic classification algorithm in this study provides quantitative reference for doctors′ diagnosis. It has the potential to improve junior radiologists′ diagnostic performance in differentiating benign and malignant breast masses.