Objective To determine the role and mechanism of peroxisome proliferator activated receptors (PPAR) α in a mouse model of D-galactosamine/lipopolysaccharide (D-GalN/LPS)-induced acute liver failure(ALF).Methods Firstly,C57BL/6 mice were randomly divided into control group(n =8),ALF 2h group(n =8),ALF 4h group (n =8),ALF 6h group (n =8).Secondly C57BL/6 mice were randomly divided into control group(n =8),ALF group(n =8),WY14643 group(n =8).To induce ALF,the mice were injected intraperitoneally with D-GalN (700 mg/kg) and LPS (10 μg/kg).WY14643 (6 mg/kg),the selective agonist of PPAR α,was administered via tail vein two hours prior to D-GalN/LPS exposure.Two,four,and six hours after D-GalN/LPS treatment in the first study,mice were anesthetized and blood was collected,6h after D-GalN/LPS treatment in the second study,blood was collected.The liver tissue was harvested for histology and mRNA extraction.Serum levels of ALT and AST were measured to evaluate the hepatic damage.Inflammatory cytokines (TNFα,IL-1β,IL-6) and chemokines (CXCL-1,CXCL-10) were detected by real-time quantitative PCR.Differential protein expression of p-NF-κBp65,p-JNK,p-ERK,p-p38 in inflammatory pathways was detected by Western blotting.Significance of inter-group differences was assessed by one-way ANOVA,and pairwise comparison was performed by the least significant difference test.Results The gene and protein expression of PPAR α were gradually reduced during the development of ALF.Compared with the model group,the liver architecture was better preserved almost with normal morphology in WY14643-treated mice.Serum ALT and AST levels in WY14643-treated group were significantly lower [ALT:(555 ±62)U/L vs (2 898 ±822) U/L,P ＜0.05; AST:(791 ±58) U/L vs (3 013 ±997)U/L,P ＜ 0.05].The expression of proinflammatory cytokines and chemokines was significantly suppressed during the activation of PPAR α.In the second study,the levels of gene expression of proinflammatory cytokines and chemokines were detected in control group,ALF group and WY14643 group respectively as followings:TNFα (0.161 ± 0.085,7.996 ± 1.068,3.346 ± 0.94,P ＜ 0.05),IL-1β(0.041 ±0.002,3.657 ±0.904,0.176±0.089,P＜0.01),IL-6 (0.018 ±0.008,1.762 ±0.589,0.163±0.0487,P ＜0.05),CXCL-1 (0.063 ±0.008,7.881 ±0.966,2.737 ±0.864,P ＜0.01),CXCL-10 (0.054 ±0.005,5.671 ±0.948,2.578 ±0.804,P ＜0.05).Conclusion Our findings first demonstrate that PPARα protects liver from injury in an ALF mouse model by suppressing inflammatory response,indicating PPARα as a potential new therapeutic target for ALF.

Objective:To establish the reference values on blood lymphocyte immunophenotype by means of standardized quality control and proper statistical analysis.Methods:The whole bloods of 93 healthy Chinese adults were stained by fluorescent conjugated monoclonal antibodies of Simultest TM IMK lymphocyte Kit and then were analysed by FACS Calibur flow cytometer.The data were acquired and were analysed by SimulSET.Statistical analysis would be conducted after conforming the date of standardized quality control.Results:CD + 3,CD + 3CD + 4,CD + 3CD + 8 cells showed normal distribution but CD - 3CD + 19 ,CD - 3CD + 16 and/or CD + 56 cells,ratio of CD + 3CD + 4 and CD + 3CD + 8 showed skewness distribution .the 95% reference of confidence interval of CD + 3?CD + 3CD + 4?CD + 3CD + 8 cells were established by the method of normal distribution and they were respectively 49 28%～80 52%?25 29%～48 11%?11 93～44 77%;the 95% reference of confidence interval of CD - 3CD + 19 ,CD - 3CD + 16 and/or CD + 56 cells,ratio of CD + 3CD + 4 and CD + 3CD + 8 were established by the method of percentiles and they were respectively 4 74%～19 84%?10 63%～42 86%?0 65～3 05.Conclusion:To establish the reference values,it is considered that the methods of normal distribution and percentiles should be use comprehensively.It is also suggested that the reference values should be confirmed by means of standardized quality control and proper statistical analysis,so the reference values obtained from various laboratories of the same race but in different regions with the same reagents and instruments performed compatibility.

Objective To analyze the types and risk factors of community-acquired infections (CAI)in diabetic patients by system analysis method of evidence-based medicine.Methods China National Knowledge Infrastructure (CNKI),Wanfang database,VIP database were searched by computer,domestic published researches on CAI and related risk factors in dia-betic patients were aggregated,Meta-analysis was conducted by stata 1 1 .0 software.Results A total of 1 2 literatures were included in the study .The average rate of CAI in diabetic patients was 39.55% (22.12%-55.86%).The major infec-tions were respiratory system infection(40.74%),urinary tract infection(27.35%),tuberculosis(10.80%),skin and soft tissue infection(9.19%),and hepatobiliary system infection (5.57%).Stratified analysis on risk factors revealed that OR and OR95%CI of chronic complication,age,disease course,glycemic control,gender,type of diabetes,subtype of ketoac-idosis was 1.63(1.45,1.82),1.30(1.19,1.42),1.47(1.35,1.61),0.68(0.61,0.76),0.69(0.64,0.75),1.37 (1.13,1.66 )and 0.87(0.62,1.23),respectively.There was no publication bias and combined results were stable. Conclusion The main CAI in diabetic patients are respiratory system infection,urinary tract infection,tuberculo-sis,skin and soft tissue infection,and so on ;several factors,such as female,older age,long-term disease course, poor glycemic control,and complication,can contribute to the increase of CAI in diabetic patients.

Objective To evaluate the plasma level of miR-21 in hepatocellular carcinoma (HCC) patients and its clinical diagnostic significance .Methods Case-control study was used .The plasma level of miR-21 was measured by real-time quantitative PCR.The relative expressions of miR-21 were calculated. This study included 60 cases of patients with HCC , 71 patients with liver cirrhosis ( LC ) , 52 healthy volunteers ( HV) from January to June in 2014 in Henan Province People′s Hospital.The receiver operating characteristic ( ROC) curves were analyzed to determine the sensitivity and specificity of miR-21 expression levels in HCC diagnosis. Differences between groups were assessed by the t-test.Results Plasma microRNA-21 level in the 60 patients with hepatocellular carcinoma ( 2.6 ±1.1 ) was significantly higher than in patients with chronic hepatitis (1.6 ±0.9) and healthy volunteers (1.0 ±0.6) (t=5.322,P=0.004;t =8.349, P =0.000 3, respectively ) .Plasma microRNA-21 level in the HCC patients were positively correlated with tumor size and differentiation (tdif=3.366,P=0.019;tsize =3.490,P=0.012). ROC analysis of plasma microRNA-21 yielded an AUC of 0.796 with 70.0% sensitivity and 65.3%specificity when differentiating hepatocellular carcinoma from chronic hepatitis , and an AUC of 0.934 with 89.5% sensitivity and 81.8% specificity when differentiating hepatocellular carcinoma from healthy volunteers.Conclusion The plasma level of miR-21 in HCC patients has high specificity , and maybe help to diagnose of HCC.

Objective To achieve the fusion expression of the entire human beta-defensin-3(hBD-3) gene. Method We synthesized two oligonucleotide primers accor ding to the codon preference of Escherichia coli. The gene was cloned into p GEX -4T-2 to establish the pGEX-4T-2-hBD-3 as the fusion expression vector by PCR. Transformed into E.coli strain DH5α, the express vector was induced an d ex pressed by IPTG. The fusion protein GST-hBD-3 was obtained by repeated cycles of freezing and thawing, cut by thrombin to attain the recombinant hBD-3 protei n. Result The result of the antibacterial peptide agarose diffu sion assay shows the antibacterial activity of the rhBD-3 against the S.aureu s exists, and it reaches 0.843U. Conclusion The fusion expr ession of the hBD-3 gene is successful.

BACKGROUND: Chronic congestive heart failure occurs at the late stage of many heart diseases. Cognitive disability exists in chronic congestive heart failure which has been reported in most clinical studies, and measures that can successfully improve heart function can improve cognitive function.OBJECTIVE: To probe into the effect of chronic congestive heart failure after myocardial infarction on spatial learning and memory ability,and to observe the correlation between heart failure and cognitive function.DESIGN: A randomized controlled observation based on the experimental animals.SETTING: Key Laboratory of Traditional Chinese Medicine (Beijing University of Chinese Medicine), Ministry of Education.MATERIALS: This experiment was performed at the Key Laboratory of Traditional Chinese Medicine (Beijing University of Chinese Medicine),Ministry of Education. in November 2001. Totally 100 healthy male rats aged 10 to 12 weeks, weighing 200 g to 220 g purchased from Beijing Weitong Lihua Experimental animal Technical Co. Ltd were enrolled and randomly divided into the operation group (n=60) and sham-operation group (n=40).METHODS: Model rats in the operation group were infarcted by ligation of the left anterior descending coronary arteries to produce acute myocardial infarction models. Those in the sham-operation group left a loose tie without ligation. The rats that showed myocardial infarction-like changes through electrocardiogram were taken as the rats for operation (n=38), and those survived rats without abnormal changes and without ligation were taken as the sham-operation group (n=25). The two groups were subdivided into 10-day gro up, 30-day group and 60-day group. Morris water maze test was performed 10, 30 and 60 days after the operation followed by hemodynamic monitoring. Among the 10-day group, 30-day group and 60-day group, rats that did not complete the whole Morris swimming test due to heart failure with poor power and died 2 days after the operation or after hemodynamic monitoring were set as heart failure end period group.Rats in the same group as those died with similar body mass were set as the control group.MAIN OUTCOME MEASURES: ① Escape logarithm latency and the good rate if cognitive ability; ② stroke volume, cardiac output per minute,heart rate and cardiac index of hemodynamics, in which cardiac index was the main index for responding heart functionRESULTS: Totally 33 rats met the criteria in the operation group and 25rats in the sham-operation group. All the animals entered the stage of result analysis. ①Results of hymodynamic index: Stroke volume, cardiac output per minute and cardiac index of the rats in the 10-day group, 30-day group and 60-day group were significantly lower than those of the homochronous sham-operation group (P ＜ 0.01), and those indexes were significantly lower in the heart failure end period group than in the end period control group and each period of the operation (P ＜ 0.01-0.05). As compared with homochronous control group, the cardiac index in 10-day group, 30-day group, 60-day group and end period group was 51.21%,50.58%,55.84% and 33.91% respectively, and all were 60% lower than the normal value. ② Results of Morris water maze: Logarithm latency of 10-day operation group after training of 8 times was obviously longer than that of the 10-day sham-operation group [F(1,28) =5.997,P=0.021]. As for cognitive function, it was obviously worse in the 10-day operation group than that of the 10-day sham-operation group (x2=8.142 ,P ＜ 0.005). There was no significant difference between the 30-day operation group and the 30-day sham-operation group, and there was no significant difference in the logarithm latency as well between the 60-day operation group and the 60-day sham-operation group. As for searching strategies, the good rate of general cognitive ability in the 60-day operation group was worse than that in the 60-day sham-operation group (x2=4.988 ,P ＜ 0.05). Logarithm latency prolonged significantly in the heart failure end period group than in the end period control group [F( 1,6)=19.567 ,P=0.004], and the good rate of general cognitive ability was worse in the heart failure end period than that in the end period control group(x=1 1.82,P ＜ 0.01).CONCLUSION: Spatial learning and memory ability after heart failure in rats with myocardial infarction was injured obviously 10 days after the operation, and it recovered to the normal level 30 days after the operation.Cognitive injury occurred to some extent 60 days after the operation, and the cognitive function was obviously worse at the end period of congestive heart failure.

Objective Study of the effects of hepatocyte growth factor on inhibit Intimal hyperplasia of the anastomotic stoma after carotid artery bypass grafting.Methods Thirty-two New Zealand white rabbits were randomly divided into control group and the experimental group.The veins were pretreated with saline solution(control group)only or pretreated with HGF(experimental group ;100ng/ml).The vein grafts were harvested at 14 days,28days after operation,HE Stain and Elastic fibrin Stain,The thickness of Intima and media in the vein grafts,intima-media ratio(I/M) was calculated by computer image analysis system.PCNA Immunohistochemistry was performed.Results The thickness of Intima and media in the vein grafts of control group surpassed experimental group significantly(P ＜0.01).At 14d I/M in the vein grafts of control group (0.81 ± 0.05) surpassed experimental group (0.47 ± 0.05) (P ＜ 0.01),At 28d I/M in the vein grafts of control group(0.73 ± 0.01)surpassed experimental group (0.65 ± 0.01) (P ＜ 0.01).The vascular smooth muscle cell proliferation in experimental group was significantly lower than that in control group (P ＜ 0.01).Conclusion Treatment of veins grafts with HGF can significantly inhibit intimal hyperplasia in a rabbit carotid artery bypass grafting model.

Objective To investigate the role of integrin-linked kinase (ILK) on migration and invasion of lung cancer cell by upregulating matrix metalloproteinase-9 through nuclear factor-κB (NF-κB) pathway.Methods A549 cell line were overexpressed ILK and matrix metalloproteinase-9 (MMP-9) confirmed by cell transfection,siRNA interference,cell scratch test,real-time quantitative PCR and Western Blot.Results Over-expression of ILK stimulated MMP-9 expression in lung cancer cells(P ＜ 0.01).The addition of MMP-9 inhibitor doxycycline and anti-MMP-9 neutralizing antibody significantly impaired the wound healing capacity of ILK-transfected cells(P ＜ 0.01),as well as by in vitro matrigel invasion assay (P ＜ 0.01).In addition,overexpression ILK induced phosphorylation and nuclear translocation of nuclear factor-κB subunit p65.Upregulation of MMP-9 was severely abolished by either BAY 11-7028,a specific NF-κB inhibitor,or siRNA targeted to NF-κB p65 in ILK over-expression cells.Conclusion The finding indicate that over-expression of ILK can promote the migration and invasion of lung cancer cell,and upregulate MMP-9 through the NF-κB pathway.

A survey on smoking status and risk factors was conducted among 3373 residents in two districts of Shen yang city during November 2008 and September 2009 ; the pulmonary function tests were also performed for all subjects.A logistical regression model was used to evaluate the risk factors of smoking,and the knowledge about diseases caused by smoking was evaluated by chi-square test.The overall smoking rate was 50.7％ ( 1710/3373 ) ; 86.8％ (1458/1680) for males a(un)14.9％ (252/1693)for females.The risk factors of smoking ( P ＜ 0.05 ) in order of OR value were as follows:drinking,work environment and underlying diseases ; the protective factors were:female,educational level,overweight ( BMI ≥ 24) and old age.The survey on knowledge of smoking-related disease was conducted in 2478 residents including 815 smokers and 1663 non-smokers. The awareness levels about whether smoking can cause COPD,asthma,birth defects,malignant tumor,abortion,growth retardation,fetal death and myocardial infarction were significantly different between smokers and non-smokers.

Objective To observe the influence of Qizhi Yifei containing serum on the expression of MMP-9 and TIMP-1 mRNA in lung fibroblasts, and explore its mechanism. Methods Trypsin digestion method was used to extract fibroblasts from lung tissue in rats. All fibroblasts were cultured and trained to the fourth generation. Then they were randomly divided into control group, model group and drug serum group. The model group and drug serum group were firstly treated by DMEM with 0.002 5 μg/mL TGF-β1. The control group was treated by DMEM only. The control group and model group were then treated by DMEM with blank drug serum in concentration of 5%, and the drug serum group was treated by DEME with Qiahi Yifei containing serum in same concentration. After 48 and 72 hours, RT-PCR was performed to test the expression of MMP-9 mRNA and TIMP-1 mRNA of each group. Results After 48 hours, MMP-9 and TIMP-1 mRNA were significantly increased in model group and drug serum group compared with control group. There was no difference between model group and drug serum group. After 72 hours, MMP-9 mRNA was up-regulated in model group and was decreased in drug serum group compared with control group. There was no significant difference among the three groups on the expression of TIMP-1 mRNA. Conclusion Qizhi Yifei containing serum can decrease the up-regulated expression of MMP-9 mRNA in lung fibroblasts stimulated by TGF-β1.