Objective To establish a multiplex polymerase chain reaction (M-PCR) assay to detect HBV DNA in the peripheral blood mononuclear cells(PBMCs) of chronic HB patients. Methods One pair of primer amplifying HBV genome DNA and another pair of primer amplifying HBV covalently closed circular DNA (cccDNA ) were added to one PCR reaction to detect HBV DNA in PBMCs. Results Various forms of HBVDNA including total DNA and cccDNA could be amplified simultaneously. Among the 30 chronic HB patients, both the HBVDNA and HBVcccDNA in the PBMCs of 23 patients were detected, the positive rate of which was 76.6%. The positive rate of HBV cccDNA accounted for 82.1% of total HBV DNA positive rate. Conclusion HBVDNA in the PBMCs could partially replicate. The M-PCR was successfully set up to amplify HBV genome DNA and HBV cccDNA simultaneously.

Objective To evaluate the cardiovascular disease (CVD) risk of the patients with metabolic syndrome (MS) by CVD predictive models. Methods The CVD risk of MS patients (n=637) and type 2 diabetes mellitus (T2DM)(n=259) were assessed by Framingham and ischemia CVD (ICVD) predictive models. The technique of myocardial nuclide development (MCND) was used on 121 MS patients to confirm the evaluation. Results The CVD risk of MS patients was significantly higher than that of T2DM. The proportion of CVD risk ≥ 5% in MS group was significantly higher than that in T2DM group. An elevated tendency of the proportion of CVD risk ≥ 10% appeared followed the age increasing in both MS and T2DM groups. The proportion of CVD risk ≥ 5% in male patients was higher than that in females assessed by Framingham predictive model, while an opposite result was given by ICVD predictive model. The MCND findings showed that, the more the increasing of CVD risk evaluated by Framingham predictive model,the severer the myocardial ischemia was. Conclusions The CVD risk of patients with MS is significantly higher than that of T2DM assessed by the both models. Framingham model is more susceptible to predicate CHD in MS patients than ICVD model.

Objective:To observe the correlation among cardiac function ,inflammation factor and plasma testoster‐one level in aged male patients with chronic heart failure (CHF) .Methods :A total of 91 male CHF inpatients were selected from our hospital ,including 31 cases with NYHA cardiac function class Ⅱ ,30 cases with class Ⅲ and 30 ca‐ses with class Ⅳ .Another 40 corresponding male patients without CHF were regarded as non‐CHF control group . Plasma levels of testosterone ,interleukin (IL)‐6 ,tumor necrosis factor (TNF)–α and brain natriuretic peptide (BNP) and cardiac function were compared between two groups .Results:Compared with non‐CHF control group , there were significant reductions in plasma testosterone level , P<0. 05 or 0. 01 ,and they more reduced along with NYHA cardiac function class rose in all CHF subgroups ,P<0.05 or <0.01 .Correlation analysis indicated that tes‐tosterone level was inversely correlated with IL‐6 and TNF‐α levels (r= -0.244 , -0.322 , P<0.05 both) ,and positively correlated with LVEF (r=0.322 ,P=0.01) .Conclusion:In patients with chronic heart failure ,the poo‐rer cardiac function is ,the lower plasma testosterone level is .Testosterone level measurement contributes to judging disease degree and prognosis of patients with chronic heart failure .

Objective To probe into the optimal concentration of Wnt-11 to induce the differentiation of bone marrow mesenchymal stem cells (BMMSCs)into cardiomyocyte-like cells in vitro.Methods BMMSCs were isolated from the bone marrow of SD rats using whole bone marrow culture method.After cultured for 48 h, BMMSCs of the second generation were utilized for directed induction.Based on the final concentration of Wnt-11 , BMMSCs were divided up into Group A (100 ng/mL),Group B (200 ng/mL),Group C (400 ng/mL)and Group D (blank control).After 72-hour induction,the cells were cultured in complete medium for 4 weeks while cells in Group D were cultured only in the complete medium.The morphological changes were observed under the phase contrast microscope.Surface antigen expression of BMMSCs was identified by flow cytometry.When cells were cultured for 4 weeks,the expressions of Desmin,Connexin43 and cTnI were detected by immunocytochemistry. Meanwhile, the ultrastructural changes were observed using transmission electron microscope. The mRNA expressions of cardiac transcription factors GATA-4,Nkx2.5 andα-MHC in BMMSCs were detected by RT-qPCR at 1,2 and 4 weeks after induction.Results Primary BMMSCs formed cell colonies at 2 weeks;the cells were mainly fusiform or star-shape,and a few irregularly-shaped ones were also visible.The passaged cells were larger than those of primary culture.After induction,the cells exhibited long shuttle-shape and were aligned in parallel. Flow cytometery displayed that the positive rate of the surface antigens of BMMSCs CD29,CD45,and CD90 was 97.9%,0.4% and 99.5%,respectively.When BMMSCs-induced via Wnt-11 were cultured for 4 weeks,Desmin, cTnI and Connexin43 were all positively expressed in induction groups.Whereas in the blank control group they were slightly positive or negative;the positive rate in Group B was the highest (P<0 .05 ).Transmission electron microscopy exhibited that organelles such as rough endoplasmic reticulum,mitochondria,as well as some ribosomes were visible in the cytoplasm of these cells in each induction group.In addition,myofilaments were arranged in parallel in the cytoplasm.The cells in induction groups could express GATA-4 and Nkx2 .5 in the first week,and then the expression of them decreased in the second week,but then increased in the fourth week;gene expression in induction Group B was significantly higher than in the other two induction groups (P<0 .05 ).The expression of GATA-4 and Nkx2 .5 in Group D was 1 ,α-MHC was not expressed in the four groups during the culture period. Conclusion Wnt-11 can induce the differentiation of BMMSCs into cardiomyocyte-like cells in vitro,and the optimal concentration of Wnt-11 is 200 ng/mL.

Objective: To research the effect and mechanism of how chitosan restrain fat-absorbtion, and provide references for developing a kind of nutritional and healthy fastfood. Methods: 1.In vitro digestion: Phosphomolybdic acid colorimetry; 2.In vivo experiment:240 rats were divided into four groups and fed with different diets: basal diet(Group A), high fat diet (Group B), chitosan supplemented diet (Group C), high fat and chitosan supplemented diet (Group D). The fat and chitosan content(%) of diet were A(10,0), B(60,0), C(8,20), D(34.3,14.8) respectively. Determine the blood lipids and fat output in dung, and observe the weight gain and swimming time. Results: 1.In vitro experiment showed chitosan acted on cholic acid at the best proportion 1∶8. 2.In vivo experiment showed chitosan can restrain fat-absorbtion, and increase fat output. Swimming in ice-water showed chitosan improved rats performance. Compared with group B, the blood lipids and weight of group C and D had no obvious change, but the fat-output much increased.

Objective To explore the method of primary culture and biological characteristics of aortic vascular smooth mus-cle cells (VSMC)in mice,providing experimental material for cellular and molecular scientific research of vascular disease. Methods Thoracic and abdominal aortas in mice were isolated and VSMC were obtained by using improved method of tissue piece inoculation.Digested with trypsin and passaged,VSMC were purified with differential adherence method.The conditions of cellular morphology and growth were observed under inverted phase contrast microscope,and VSMC were identified with hematoxylin-eosin (HE)staining and immunofluorescence.Results VSMC were isolated successfully and grown vigorously with good bioactivity,the cells had a radial or typicalpeak-valleylike growth,and showed fusiform,abundant cytoplasm with large and round or oval nucle-us by HE staining,the expressions of specific cytoplasmicα-smooth muscle actin were positive by immunofluorescence stain.Conclu-sion It can isolate and cultivate VSMC with high purity and good activity under in vitro conditions with simple,economical,reliable method.

Music electrotherapy is a fusion of music therapy and electrotherapy,as a new digital medical technique,it has been concerned by medical workers.In this paper,a portable music therapeutic apparatus based on ARM-WINCE was developed.This apparatus could offer electrical stimulation,audio signal and visual signal provided by LCD.The core of system was ARM9 processor S3C2440,and using Windows CE as GUI development tool.The therapeutic apparatus has been developed and passed the detection.It has the features of small size,high reliability,low cost,and strong alternating capability,which is optimum in the fields of home health care and clinical research.

Objective To analyze the usual death causes of cardiovascular disease and the differences in gender and age .Methods By adopting the retrospective study method ,the clinical data of death cases in cardiovascular disease were collected and analyze on the situation suffering from cardiovascular disease ,direct death causes ,gender and age difference .Results (1) among 181 cases of cardiovascular disease death ,coronary heart disease(115/181 ,64% ) and hypertension(96/181 ,53% ) were the most common dis-ease ,lung infection(104/181 ,57% ) was the most common complication ;(2)There was no significant difference in the situation suf-fering from basic diseases between male and female(P>0 .05);(3)The basic diseases in cardiovascular death cases aged over 60 years old were dominated by coronary heart disease and hypertension ;the proportion of complicating pulmonary infection was grad-ually increased with age increase ;(4) in the direct death causes ,the top 3 places were sudden cardiac death (44/181 ,24 .3% ) ,multi-ple organ dysfunction syndrome(24/181 ,13 .3% ) and cardiogenic shock(24/181 ,13 .3% ) .Conclusion Strengthening the manage-ment of diagnosis and treatment on elderly patients with coronary heart disease ,hypertension ,especially those complicating diabe-tes ,strengthening the treatment intervention of lung infection in cardiovascular disease population and conducting the emphasis pro-tection on the target organ function may reduce the mortality of cardiovascular inpatients .

Objective To observe the effects of Slit2 protein on the proliferation and migration of VSMCs .Methods The VSMCs was cultured in our laboratory .The experiment was divided into two parts ,part one:VSMCs were divided into normal con‐trol group and experimental groups(culture with 50 ,75 ,100 ,125 and 150 ng/mL Slit2 respectively);part two :VSMCs were divided into normal control group ,positive control group(culture with TNF‐α10 ng/mL) and experimental groups(culture with TNF‐α10 ng/mL+Slit2 50 ng/mL ,TNF‐α10 ng/mL+Slit2 75ng/mL ,TNF‐α10 ng/mL+ Slit2 100 ng/mL ,TNF‐α10 ng/mL+ Slit2 125 ng/mL and TNF‐α10 ng/mL+Slit2 150 ng/mL respectively) .To detect proliferation and migration of VSMCs by CCK‐8 and tran‐swell experiment .Results The difference of OD value and numbers of VSMCs has no statistical significance in the presence of Slit 2 (P=0 .516 ,P=0 .52) .The numbers of VSMCs has statistical significance between control and positive control groups (P=0 .00) . The numbers of VSMCs in experimental groups were fewer than positive control group (P<0 .05) ,whereas the difference of OD value still has no statistical significance between experimental and positive groups (P= 0 .173) .Conclusion Recombinant Slit2 could inhibits migration in VSMCs induced by TNF‐α,whereas it has no effect on proliferation of VSMCs .

Objective To investigate whether thalidomide inhibits the over expression of type I collagen in pulmonary fibrosis rats via inhibiting the JNK signaling pathway,thereby reducing bleomycin induced pulmonary interstitial fibrosis in rats.Methods 90 healthy male SD rats were randomly divided into normal control group (group N),model group (group M),thalidomide group (group T),SP600125 group (group SP) and thalidomide+SP600125 group (group T+SP).The pulmonary fibrosis models were prepared via intratracheal injection of 5mg/kg bleomycin,and rats in groups were given corresponding drugs from the first day after preparing model.Rats were randomly sacrificed at 7,14 and 28 days after treatment.The degree of pulmonary alveolitis and fibrosis was evaluated by H&E and trichrome masson stainings.The level of hydroxyproline in the lung tissue was detected by applying alkaline hydrolysis technique,and expression levels of p-JNK and type I collagen were tested by Western bloting for protein expression and real-time polymerase chain reaction (RT-PCR) for mRNA expression.Results In group M,alveolitis was the most serious on day 7; a marked pulmonary fibrosis formed on day 28; the level of hydroxyproline also peaked on day 28,and the contents of p-JNK and type I collagen were higher than in group N(F=277.87,472.51,both P＜ 0.01).Group T,SP and T+SP showed mild alveolitis and fibrosis at all time points,and their levels of hydroxyproline,p-JNK and type I collagen were remarkably decreased as compared with group M (F=14.77,61.59,101.73,all P＜0.01;F=10.33、79.12、57.48,all P＜0.01).No significant difference in p JNK was found between group SP and group T+SP.Conclusions Thalidomide may inhibit the over expression of type I collagen in pulmonary fibrosis rats via inhibiting the JNK signaling pathway,thereby reducing bleomycin induced pulmonary interstitial fibrosis in rats.