Objective To study the changes and significance of sCD14 in inflammatory response of patients with gouty arthritis.Methods CD14 mRNA was measured using quantitative real-time PCR in peripheral blood mononuclear cells (PBMCs).The expression of CD14 mRNA in PBMCs was compared between patients with acute gouty arthritis (AGA)(n =31)and non-acute gouty arthritis (NAGA)(n =23)and healthy controls (HC)(n =20).β-actin was selected as the internal control.The protein expressions of sCD14,IL-1βand TNF-αwere measured using enzyme linked immunosorbent assay (ELISA) in patients’ plasma.The protein expression of CRP was measured using immunoturbidimetry in patients’ plasma. Routine blood and blood biochemistry indexes were measured by routine blood analyzer and blood biochemistry analyzer of patients with AGA,NAGA and HC.We analyzed the correlation between CD14 mRNA,sCD14 protein expression and each clinical indicator.Results When compared with that in AGA group,the mRNA expression of CD14 increased significantly in PBMCs of HC patients (P < 0.05 ).When compared with that in HC and NAGA patients,the protein expression of sCD14 increased significantly in the plasma of AGA patients (P <0.01).The protein expression of sCD14 was significantly lower in the plasma of NAGA than in HC (P <0.05).The protein expression of sCD14 increased significantly in the plasma of AGA compared with HC and NAGA (P < 0.01 ).When compared with those in HC,the protein expressions of IL-1β and TNF-α increased significantly in the plasma of AGA and NAGA (P < 0.01 ).When compared with that in NAGA,the protein expression of IL-1βincreased significantly in plasma of AGA (P <0.01). The indexes of WBC increased significantly in AGA compared with HC (P <0.01),and WBC increased significantly in NAGA compared with HC (P <0.05).The indexes of GR and MO increased significantly in AGA compared with HC (P <0.05),and MO increased significantly in AGA compared with NAGA (P < 0.05 ).The indexes of UA increased significantly in AGA and NAGA compared with HC (P <0.01).There was a positive correlation between CD14 mRNA expression and IL-1β in PBMCs in AGA group (r s =0.362,P =0.045).A positive correlation was found between GR and the protein expression of sCD14 in NAGA patients’plasma (r s = 0.397,P = 0.030 ). Conclusion The dysregulated expressions of CD14 mRNA in PBMCs and sCD14 protein in GA show that sCD14 may play a significantly regulatory role in inflammatory reaction.

Objective To investigate the effect of edaravone plus beating-heart-preservation as well as beating-heart-transplantation technique on myocardial protection in donation after cardiac death (DCD) heart transplantation.Methods Twenty-four swine (body weight 28 ± 3 kg) were divided into two groups (n =12 each),and another twelve swine were used for blood donor.( 1 ) Experimental group:cardiac arrest was induced by asphyxiation (turning off the ventilation),and then the swine were subjected to 25-min warm ischemia,and cold oxygenated blood was perfused before the harvest of donor heart.Cardiac resuscitation was initiated by the ex vivo perfusion equipment and warm oxygenated blood was reperfused.Edaravone was given before harvesting of donor heart and in the early period of reperfusion.Donor hearts were kept beating throughout preservation and transplantation period.(2) Control group:all animals were treated in the same way except for without the application of edaravone.Hemodynamic,myocardial enzymes,and water content of myocardium were observed,and uhrastructural damage of cardiomyocytes was examined.Results All recipient animals could wean from cardiopulmonary bypass successfully.Left ventricular compliance and left ventricular contractility were significantly better preserved in experiment group than in control group.Though there was no significant difference in myocardial creatase level,the myocardial edema in experimental group was milder than in control group,and myocardial ultrastructure was better preserved in experimental group.Conclusion Heart from DCD,even though experienced 25-min warm ischemia after cardiac arrest by asphyxiation,still could be resuscitated via isolated heart perfusion equipment ( i.e.,beating-heart-preservation ) successfully.Furthermore,beating-hearttransplantation is feasible technically.Edaravone,a free-radical scavenger,could alleviate asphyxiation-induced myocardial injury,and further improve post-transplantation heart function.

Objective To study the expression level of NLRP3 (NLR family, pyrin domain containing3) inflammasome (NLRP3, ASC, caspase-1 ) mRNA in peripheral blood monocytes (PBMCs) from patients with gout arthritis (GA) and to explore the pathogenesis of GA. Methods NLRP3 inflammasome mRNA was measured using quantitative real-time PCR in PBMCs. The expression of NLRP3 inflammasome mRNA in PBMCs was compared between patients with GA (n=24) and healthy controls (n=24). β-actin was selected as the internal control. Students' t-test was used in two independent samples and Spearman's correlation was used to evaluate the relationship between mRNA level and inflammatory parameters. Results The expression of ASC mRNA in GA increased significantly when compared to healthy controls [(0.029±0.021 ) vs (0.009±0.007 ), P＜0.01 ], and the expression of NLRP3 and caspase-1 mRNA was significantly lower in patients with GA compared to healthy controls [(0.062±0.084) vs (0.133±0.106), P＜0.05; (0.025±0.014) vs (0.117±0.156), P＜0.01]. Moreover, the expression of ASC mRNA was found to correlate significantly with globulin (r=-0.547, P＜0.05) and very low density lipoprotein cholesterol (r=-0.540, P＜0.05) in GA patients,and caspase-1 was correlated to globulin(r= -0.773. P＜0.01) and very low density lipoprotein cholesterol (r=-0.465. P＜0.05). Furthermore, the ASC mRNA in GA patients was associated significantly with NLRP3 mRNA(r=-0.450, P＜0.05) and caspase-1 (r=0.604, P＜0.01 ). Conclusion Dysregulated expression of the NLRP3inflammasome is involved in the inflammatory response and plays a key role in the pathogenesis of GA.

Objective To investigate the single nucleotide polymorphisms(SNPs) rs3733591(C>T) of SLC2A9 gene in Chinese Han population, and to explore the association of this gene polymorphisms with gout susceptibility, tophi, serum uric acid levels, other clinical and laboratory data and the levels of SLC2A9 mRNA of peripheral blood mononuclear cells(PBMCs). Methods ① A total of 297 primary gout arthritis patients(GA) and 211 normal controls(NC) were enrolled into this study. The clinical and laboratory data of patients were collected. The genotypes and alleles frequencies were measured by using TaqMan ?SNP Geno-typing Assays and the possible association between gene polymorphism of SLC2A9 and gout was investigated by Chi-square test. The odds ratios(OR) and 95% confidence intervals(95%CI) were calculated. ② The lev-els of SLC2A9 mRNA on PBMCs of 86 gout patients(46 patients in remission) and controls were measured by real-time quantitative polymerase chain reaction (RT-qPCR). The nonparametric test was used to analyze the expression in different groups. Results The frequencies of genotypes and alleles of rs3733591(C>T) in gout patients were different from controls(P<0.05). The frequency of TT genotype was significantly lower than that in controls (P<0.05) and the relative risk of this genotype to develop gout was 0.647 (95%CI: 0.452-0.925). Moreover, the frequency of T allele in cases was much lower than in controls (60.9% vs 69.2%, χ2=7.324, P=0.007, OR=0.695), but the frequency of C allele was much higher(39.1% vs 30.8%, χ2=1.440, P=0.007, OR=1.440). Interestingly, the levels of SLC2A9 mRNA on PBMCs in gout patients who carried TC genotype of rs3733591 was higher than those who carried TT genotype(P<0.05). There was no difference in the expression of SLC2A9 mRNA on PBMCs among different genotype carriers of rs3733591 in controls (P>0.05). However, there was no significant difference in the distribution of genotypes and alleles between 30 tophaceous gout patients and 190 non-tophaceous gout patients(P>0.05). Conclusion Results of present study suggest the rs3733591(C>T) polymorphism of the SLC2A9 gene might be associated with gout development, but not with tophaceous gout. The C allele predisposes to gout, and TT genotype and T allele might protect Chinese Han population from developing gout. The rs3733591(C>T) polymorphism probably affects the susceptibility to gout by influencing the f expression of SLC2A9 mRNA susceptibility.

Objective This study is aimed to evaluate the association between the ABCG2 gene rs2231142 variant and gout using meta-analysis. Methods Related studies were identified by searching extensively in Chinese and foreign language databases such as Pubmed, EMBASE, Cochrane Library, CBMdisc databases and so on. The quality of included studies was assessed by using the Newcastle-Ottawa Scale (NOS). The odds ratio (OR) was calculated using a random-effects or fixed-effects model. A Q statistic was used to evaluate the heterogeneity, and Eggerˊs test and funnel plot were used to assess publication bias. Sub-group analyses on ethnicities and sex were also performed. Results A total of 10 studies, including 3 478 gout patients and 10,089 controls from 6 countries or regions, were included and identified for the current metaan-alysis. It was found that the A allele or AA genotype of the ABCG2 rs2231142 polymorphism had an increased risk for gout in the general population [A allele: OR=2.03, 95%CI (1.77, 2.34), P<0.01 and AA genotype: OR=3.01, 95%CI (2.34, 3.88), P<0.01, respectively]. Similar results were found in sub-group analyses of different gender and races. Conclusion Existing evidence indicate that rs2231142 polymorphism (the A allele and AA genotype) is associated with increased risk of gout.

Objective To investigate the role of high mobility group box 1 protein(HMGB1) and the receptor for advanced glycation end products (RAGE) in the pathogenesis of primary gouty arthritis (GA).Methods Enzyme-linked immunosorbent assay(ELISA) was used to determine the level of plasma HMGB1 in 68 acute gout (AG),48 quiescent gout (QG) and 45 healthy control(HC).Real-time quantitative polymerase chain reaction (RT-qPCR) was employed to measure the expression of HMGB1 and RAGE mRNA in the peripheral blood mononuclear cells (PBMCs) in 68 AG,48 QG and 94 HC.One way ANOVA or Wilcoxon test and Spearman's correlations were used for statistical analysis.Results The level of plasma HMGB1,PBMCs HMGB1 and RAGE mRNA were significantly higher in GA than that in HC [(24±34) ng/ml,0.019±0.029,0.000 5±0.000 3] (P＜0.05),while the level of plasma HMGB1 and PBMCs HMGB1 mRNA were significantly higher in AG [(222±178) ng/ml,0.235±0.954,0.001 5±0.003 5] than that in QG [(107±176) ng/ml,0.044±0.117,0.001 3±0.000 9] (P＜0.05),and the level of PBMCs RAGE mRNA was higher in AG than that in QG (P＞0.05).In the GA patients,the level of plasma HMGB1 was positively correlated with white blood cell count,neutrophile granulocytes count,mononuclear cells and erythrocyte sedimentation rate (r=0.34,0.44,0.39,0.33; P＜0.05),while negatively correlated with apolipoprotein A1 (r=-0.28,P＜0.05); the level of PBMCs HMGB1 mRNA was positively correlated with RAGE mRNA,white blood cell counts,neutrophil counts,lymphocyte counts,serum total cholesterol level,low density lipoprotein level and apolipoprotein B100 level (r=0.29,0.36,0.26,0.28,0.29; P＜0.05),while negatively correlated with high density lipoprotein (r=-0.30,P＜0.01); the level of PBMCs RAGE mRNA was positively correlated with lymphocyte counts,total cholesterol and apolipoprotein B100 (r=0.35,0.35,0.44; P＜0.05).Conclusion HMGB1 and its signaling pathway may play important role in the pathogenesis of gouty arthritis,which may also be involved in the regulation of the lipid metabolism of gout.

Objective To investigate the role of adiponectin (ADP) and its receptors (ADR) in pati ents with primary gouty arthritis (GA).Methods Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of plasma ADP in 88 GA and 80 healthy controls (NC).Real time quantitative polymerase chain reaction (RT-qPCR) was employed to study the expression of ADR1 and ADR2 mRNA in peripheral blood mononuclear cells (PBMCs).The biochemical indicator of TG,HDL,LDL,VLDL,apoA1,apoB100 and uric acid (UA) were detected at the same time.T test,Spearman's correlations and regression analysis were used for statistical analysis.Results The concentration of plasma ADP was significantly lower in GA than that in NC [(6±7) μg/ml,(8±6) μg/ml,t=-3.71,P＜0.01 ],the expression of ADR1 and ADR2 mRNA in GA (ADR1:0.09±0.08,ADR2:0.0122±0.0164) was significantly increased when compared to the NC (ADR1:0.05±0.03,ADR2:0.0054±0.0024) (t=2.71,2.35,P＜0.05).In the GA patients,the level of ADP was negatively correlated with the lymphocytes count (LY) and UA (r=-0.32,-0.36,P＜0.05),and it was positively correlated with ESR and LDL level (r=0.31,0.39,P＜0.05).The expression of ADR1 mRNAwas negatively correlated with TG (r=-0.43,P＜0.05 ),but positivdy correlated with ESR and CRP level (r=0.45,0.57,P＜0.05).The expression of ADR2 mRNA was negatively correlated with glucose and UA(r=-0.50,-0.59,P＜0.05).Conclusion Altered expression of ADP and its receptors may be involved in thepathogenesis of gouty inflammation.

Objective To explore the therapeutic effect of Jinkoubao on oral ulcer in rabbits and its action mechanism.Methods Among 60 SPF NewZealand rabbits,6 cases were randomly selected for the ulcer model identification,and the rest was randomly and equally divided into 3 groups:the control group (NC group),normal saline drug film group (NS group) and Jinkoubao film group (JK group).The rabbit model of oral ulcer was constructed by applying 40 ％ glacial acetic acid to burn rabbit oral mucosa for duplicating the oral ulcer model.of the change situation of oral ulcer was observed on the same day for constructing model,on 3,5,7 d after medication.The EGF level in oral mucosal tissue was detected by using RT-PCR and the local histopatho1ogical changes in oral ulcer was observed by using HE staining method.Results Compared with the NS group,the ulcer area on 3,5,7 d after medication in the JK group was significantly deceased (P＜0.01).Compared with the NS group,the EGF level in oral buccal mucosal tissue in the NS group and JK group was markedly increased (P＜0.01).But the EGF level increase in the JK group was faster than that in the NS group,the difference was statistically significant(P＜0.01).The HE staining section of rabbit oral ulcer on 3,5,7 d after medication showed that the inflammatory cells decrease in the JK group was more obvious than that in the NS group,fibroblasts proliferation was obvious and epithelial hyperplasia was good.Conclusion Jinkoubao could greatly improve the symptoms of oral ulcer in rabbits and promotes its healing,which is possible to strengthen the repair capacity of oral ulcer by regulating the EGF level.

Objective To detect the distribution of SLC2A9 rs10489070 polymorphism genotypes in Chinese Han population,and to explore the association of this gene polymorphism with gout susceptibility,tophi,serum uric acid levels and other clinical and laboratory data.Methods A total of 151 primary gout patients and 176.healthy controls were enrolled into this study.The genotypes and alleles frequencies were calculated by using TaqMan(R) SNP Genotyping Assays and the possible association between gene polymorphism of SLC2A9 and gout was investigated.T test,Chi-square and Fisher exact probabilities were used for statistcal analysis.Results Genotypes distribution were in Hardy-Weinberg equilibrium in gout patients and controls (P＞0.05).The frequency of CC genotype in gout patients was significantly higher than that in the controls (78.8％ vs 68.5％,P＜0.05),aand the frequency of CG genotype in gout patients was significantly lower (19.9％ vs 30.1％,P＜0.05).However,there were no statistical differences in the alleles frequencies of C and G between gout patients and controls (P＞0.05).Interestingly,there was significant difference in the distribution of genotypes between tophaceous gout patients and non-tophaceous gout patients (P＜0.05),and the frequency of CG genolype was much lower in tophaceous gout patients (0 vs 22.7％,P＜0.05).Conclusion Results of present study suggest that rs10489070 polymorphism of the SLC2A9 gene might be associated with gout development.CC genotype predisposes to gout,and CG genotype might protect Chinese Han population from gout and tophi development.

objective The roles of TLRs and their signal pathway in gouty arthritis(GA)were explored.Methods TLR2 and TLR4 mRNA was measured using real-time quantitative polymerase chain reaction(RT-PCR)in PBMCs,IL-1β level was detected using ELISA in plasma,and NF-κB p65 protein level in PBMCs was measured using Western blot.Level of TLR2 mRNA,ILR4 mRNA,IL-1β,NF-κB p65protein was compared among acute GA,non-acute GA and healthy controls.Correlation between TLR2mRNA,TLR4 mRNA and serum uric acid,IL-1β level in GA patients was analyzed.One-way ANOVA was used to analyze data between multiple groups and q-test was used for two-two comparison.Spearman's analysis was applied for correlation analysis.Resuits The expression of TLR4 mRNA,NF-KB p65 protein,IL-1β arid serum uric acid level in patients with acute GA [(5.0±1.2), (7.11±0.18), (283±83)pg/ml,[585±123)μmol/L] was significantly increased compared to non-acute GA[(2.3±0.4),(0.63±0.06),(134±29)pg/ml,(493±107)μmol/Lj and healthy controls(1.1±0.6),(0.52±0.12),(97±17)pg/ml,(326±65)μmol/L](P<0.01,respectively).Significant diffefence was also observed between non-acute GA patients and healthy controls(P<0.05,respectively).The level of TLRR4 mRNA was positively correlated with uric acid and IL-1β level in GA patients(rs=0.876,0.779;P<0.05,respectively).Conclusion Innate immunity are activated by membrane-type pattern recognition receptors in primary GA.TLR4-NFκB p65-IL-1β signat transduction may participate in the inflammatory mechanisms of gout.Urate crystals in patients with gout may:be involved in the activation of TLR4 and its signal pathway.