Multiple risk factors lead to occurrence of cardiovascular diseases in common.Besides confirmed tradition-al risk factors,such as diabetes mellitus,dyslipidemia,smoking and hypertension etc.,some new risk factors have been found in recent years.The confirmed risk factors need further research,while the new risk factors need en-hanced study.

BACKGROUND:Tissue engineering requires a lot of seed cells. Osteoblasts have become important seed cells in bone tissue engineering. However, it is difficult to culture the osteoblasts, and cellnumber, purity, proliferation and differentiation activity are different obtained by different culture methods. OBJECTIVE:To identify and compare three common primary osteoblat culture methods, and to explore a method for the primary culture of osteoblasts which is easy to operate, economical and effective, in order to provide basis for the further experimental research. METHODS:Calvarias were dissected from newborn Sprague Dawley rats in 72 hours, and osteoblasts were isolated with col agenase digestion method, sequential digestion method and bone tissue method respectively. The morphological observation and cytochemical staining were performed, the growth curve of the cells was drawn with cellCounting Kit-8 method, and the rate of living osteobalsts was counted with trypan blue staining. RESULTS AND CONCLUSION:The proliferation of the insolated and cultured osteoblasts was wel with typical characteristics of osteoblasts, cytochemical staining results were positive. Compared with the sequential col agenase digestion method, the col agenase digestion method presented higher production of osteoblasts and higher cellsurvival rate (P<0.05), and the col agenase digestion method was easier than the sequential col agenase digestion method and cost less than sequential col agenase digestion method. Bone tissue method was the easiest method with less damage to cells, but bone tissue method presented lower production of osteoblasts and cost much more time, which cannot be used in large-scale osteoblast culture. The col agenase digestion method is a simple, efficient and ideal method for isolation and culture of primary osteoblasts.

Objective: To investigate the effect of Tanreqing injections combined with azithromycin in the treatment of children bronchial pneumonia. Methods:Totally 56 cases of pediatric bronchial pneumonia were selected in our hospital from September 2012 to September 2013 and randomly divided into the observation group and the control group with 28 cases in each. The control group was with azithromycin treatment, and the observation group was with Tanreqing injections additionally. The effect and adverse reactions of the two groups were studied and compared. Results:In the observation group, the effective rate was 92. 9%, which was significantly higher than that in the control group(P<0. 05). The hospitalization time and the symptoms disappearance time in the observation group were significantly shorter than those in the control group (P<0. 05). The adverse reactions showed no significant difference be-tween the two groups. Conclusion:Tanreqing injections in the treatment of children bronchial pneumonia is rapid, effective and safe.

Objective To determine the cleaning and the shortest disinfectant time for gastroscope Methods 1? g/ml HBsAg and 3? 108/ml bacteria of Aureus Staphylococci(ATCC 6538),Escherichia coli(ATTC 8099),Bacterium earuginosum(ATCC 27853)were artificially spreaded on the body of gastroscope ,biopsy hole and biopsy clamp.The gastroscope was routinely cleaned and disinfected.HBsAg and pathogens in the samples,which collected before and after cleaning the gastroscope and 3min,5min,10min after disinfection,were tested by using ELISA and bacteria culture,respectively.Results Pathogen and HBsAg can be detected before cleaning the gastroscope.Pathogen can also be detected,but HBsAg disappears after cleaning.Both HBsAg and pathogen was negative 3 min after gastroscope was immersed in glutaraldehyde.Conclusion The tests for pathogens and HBsAg can be negative after gastroscope was cleaned and immersed in 2％ glutaraldehyde for 3 min.

ObjectiveTo investigate the protective effects of p38MAPK inhibitor SB203580 on acute necrotizing pancreatitis associated lung injuries in rats.Methods Fifty-four SD male rates were randomly divided into 3 groups,including control group,ANP group,SB203580 group with 18 rats in each group.ANP was induced by intraperitoneal injection of L-arginine solution. Rats in control group were intraperitoneally injected with same amount of saline.Before ANP induction,the rats in SB203580 group received 10 μmol/L SB203580 dissolved by dimethyl sulfoxide at a dose of 5mg/kg weight via intraperitoneal injection.The rats were sacrificed at 3,6,and 12 h after operation,the serum levels of amylase,TNF-α,IL-6 was determined.Pathological changes of pancreas and lung were observed.The wet/dry (W/D) weight ratio of lung and MPO were measured.CINC mRNA of lungs was determined by RT PCR. Expression of phosphated-p38MAPK (p-p38MAPK) protein was evaluated by Western blotting.ResultsThe serum levels of amylase,TNF-α,IL-6and wet/dry (W/D) weight ratio of lung,MPO activity,CINC mRNA and p-p38MAPK protein expression of lungs were (1035 ±73)U/L,(0.94 ±0.16)μg/L,(4.77 ±0.86) μg/L,3.92 ±0.29,(0.39 ±0.02)U/g,0.28 ±0.04,0.09 ±0.04 in control group at 6 h after operation,and the corresponding values were (5848 ±656) U/L,(3.84 ±0.32)μg/L,(103.54 ± 15.32)μg/L,4.97 ±0.47,(1.03 ±0.08) U/g,0.62 ±0.06,0.52 ±0.14 in ANP group,while they were (4259 ±286) U/L,( 1.64 ±0.21 ) μg/L,(76.56 ± 11.46) μg/L,4.32 ±0.34,(0.78 ±0.05)U/g,0.37 ±0.04,0.27 ±0.08 in SB203580 group.The values in ANP group were significantly higher than those in control group,and the values in SB203580 group were significantly lower than those in ANP group,but they were still significantly higher than those in control group ( P ＜ 0.01 ).ConclusionsSB203580 may attenuate injury of lung and pancreas in ANP by blocking p38MAPK signal transduction pathway,and decreasing the production of inflammatory cytokines.

Objective To explore the potential role of monocyte chemotactic protein-1 ( MCP-1 ) gene in the pathogenesis of acute lung injury (ALI) in early acute necrotizing pancreatitis (ANP). Methods Forty SD rats were randomly divided into control group, ANP 3, 6, 12 h group with 10 rats in each group according to a number table. ANP was induced by intraperitoneal injection of 15% L-arginine solution at a dose of 2.0 mg/g body weight. Pathological changes of pancreases and lungs were observed. Lung wet/dry weight ratio was measured. Intrapulmonary expression of MCP-1 mRNA was evaluated by RT-PCR. Results After intraperitoneal injection of 15% L-arginine solution, the rat's pancreas presented with bleeding, necrosis comparable with pathological changes of ANP. Pulmonary tissue edema was obvious. At ANP 3, 6, 12 h group, the pathological scores of the lung were 3.75±0.58,5.50 ±0.63,5.86 ±0.54, the wet/dry weight ratios were 4.85 ±0.38,4.97 ± 0.47,5.03 ± 0. 46, the MCP-1 mRNA expressions were 0.36 ± 0.08, 0. 56 ± 0. 15, 0. 72 ± 0.21,which were significantly higher than those in the control group (0.12 ±0.05,4.32 ±0.33,0.21 ±0.05, P＜0.05 or ＜0.01 ). The MCP-1 mRNA expression in lungs was significantly correlated with the degree of lung damage and wet/dry weight ratio of lungs (r=0.75,r=0.89,P＜0.05).Conclusions MCP-1 mRNA expression was up-regulated in the early phase of ANP in the lungs, and it may play an important role in ALI during ANP.

Objective To investigate the proportion of CD4+CD25highTreg and CD4+CD25+CD127low/-Treg in peripheral blood in patients with chronic hepatitis B(CHB) and determine the relationship between the proportion of CD4+CD25+Treg and clinical parameters.Methods Fresh isolated peripheral blood mononuclear cells(PBMCs) of 28 patients with CHB and 24 healthy donors were analyzed for the proportion of CD4+CD25+Treg using flow cytometry by surface staining for CD4-PC5,CD25-FITC,CD127-PE.HBsAg,HBsAb,HBeAg,HBeAb and HBcAb were evaluated.HBV DNA levels were measured using real-time RT-PCR.Results The proportions of CD4+CD25highTreg to CD4+ T cells(3.36%?2.59%) and CD4+CD25+CD127low/-Treg(4.05%?1.63%) to CD4+ T cells in patients with CHB were higher than those in health controls(1.60%?0.66%,1.75%?0.83%,P100U?L-1) had a higher fraction(4.26%?3.10%) of CD4+CD25highTreg in peripheral blood than those patients with low level ALT(ALT

In total, 106 patients with maintenance hemodialysis (MHD) were divided into two groups based on their valsartan administration, and 53 healthy controls were recruited in the study. Plasma concentrations of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), resistin and high sensitivity C-reactive protein (hs-CRP) were measured by enzyme-linked immunosorbent assay (ELISA) before and after valsartan treatment. Plasma levels of resistin, IL-6, TNF-α and hs-CRP were all significantly increased in patients with MHD as compared to those in healthy controls (P＜0. 05). Among patients with MHD the plasma levels of resistin, IL-6 and hs-CRP in valsartan group were lower than those in non-valsartan group (P＜0. 05).

Aim To investigate the effects of bone morphogenic protein(BMP)-7 on the expression of extracelluar matrix components(ColⅠand FN)in human renal proximal tubular cells(HK-2)induced by transforming growth factor-?1(TGF-?1),and to explore the inhibition of BMP-7 on renal interstitial fibrosis.Methods HK-2 cells were treated with TGF-?1,BMP-7,or a combination of both for 48 h.The expression of FN was assessed by indirect immunofluorescence.RT-PCR and Western blot were used to determine the mRNA and protein expression of FN and ColⅠ?1.Results Indirect immunofluorescence staining showed that FN staining in 3 ?g?L-1 TGF-?1 group was considerably stronger than that in control group.By an addition of 200 ?g?L-1 BMP-7,the expression of FN was significantly inhibited.RT-PCR and Western blot showed the mRNA and protein expression of FN and ColⅠ?1 were up-regulated by TGF-?1 after 48 hours significantly(P

Aim To investigate the pharmacokinetics of dipfluzine hydrochloride,a novel piperazines calcium antagonist.Methods Eighteen Beagle dogs were randomly divided into three groups,which were administered with dipfluzine hydrochloride at iv single dose of 1.5,3.0 and 6.0 mg?kg-1,respectively.The blood was collected at different time.A RP-HPLC method was developed to determine the concentration of dipfluzine hydrochloride in plasma.The pharmacokinetic parameters were calculated by 3P97 software.Results The specificity,lowest limit of detection and quantification,extraction recoveries,the precision of intra-and inter-day and stability were qualified to the pharmacokinetic study.The concentration-time courses of dipfluzine hydrochloride were best fitted to a two-compartment open model at three doses.The main pharmacokinetic parameters at three doses were 24.7,24.2 and 29.6 h for T12?,0.44,1.12 and 2.86 g?min?L-1 for AUC,1.30,1.22 and 1.28 L?kg-1 for Vc,and 3.4?10-3,2.7?10-3 and 2.1?10-3 L?kg-1?min-1 for CL,respectively.Conclusions The developed RP-HPLC method for determination of dipfluzine hydrochloride in plasma can satisfy the requirement of pharmacokinetic study after iv dipfluzine hydrochloride.Analysis of plasma concentration-time curves indicates a biphasic decrease.There was a linear relationship between AUC and dosage.