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Objective To investigate the effect and mechanism of lipoxin A4 (LXA4) on uric acid (UA) induced oxidative stress of human umbilical vein endothelial cells (HUVECs).Methods The HUVECs was treated with uric acid to constructing the model of oxidative stress,and intervene the model with LXA4 and xylene based iodine (DPI),rotenone.Reactive oxygen species (ROS) of HUVEC were detected by a fluorescence probe 2',7'-dichlorofluorescin diacetate (DCFH-DA).The activity of reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and p47phox protein was measured by Lucigenin enhanced chemiluminescence and Western blotting among control,uric acid (UA),LXA4 and UA +LXA4 groups,respectively.All the results were described by the relative expression of the control group,repeated measure variance analysis and least significant difference test (LSD) were used for statistical analysis.Results UA could stimulate HUVEC to generate ROS with different concentrations and times (F=7.286,F=4.532,P＜0.05).Compared with the control group(100±l 1),the ROS production of group with 80 mg/L UA (177±18),120 mg/L (226±29) and 160 mg/L (225±16) increased significantly (t=4.127,t=7.591,t=7.236,P＜0.05).Compare with baseline(100±8),the ROS production increased significantly (t=3.688,t=3.513,t=4.526,t=8.269,t=3.829,P＜ 0.05) at 3 h(143±16),6 h(140±17),12 h(183±20),24 h(240±29) and 48 h(160±22).LXA4 could inhibit ROS generation at different concentrations and times (F=4.008,F =4.497,P ＜0.05).Compared with LXA4 concentration of 0 nmol/L,the LXA4 concentrations of 10 nmol/L (162±16) and 100 nmol/L (132±15) could significantly inhibit ROS generation(t=3.712,t=4.083,P＜0.05).Compared with pretreatment (269±39),the ROS generation decreased significantly (t=6.373,t=6.426,t=7.125,t=6.981,P＜0.05).with LXA4 pretreated for 15 min (160±16),30 rain(158±21),1 h (136±13) and 2 h(140±13).Compared with the UA group(252±31),LXA4 and DPI could significantly inhibited ROS generation (145±29,154±27;t=6.356,t=5.853,P＜0.05),but Rot was not significantly intervented (241±32;t=1.027,P＞0.05).The NADPH oxidase activity in the UA group was significantly higher than that in the control group (144±16,100±13;t=3.659,P＜0.05),but the group of LXA4+ UA was significantly lower than that of the UA group (119±14;t=3.124,P＜0.05).The cytoplasmic expression of NADPH oxidase subunit p47phox of UA group was significantly lower than that in the control group (47±6,100±8;t=7.562,P＜0.05),but the LXA4+UA group was significantly increased compare with the UA group (83±6,t=5.386,P＜0.05).The cytomembrane expression of p47phox of UA group was significantly higher than that in the control group (328±36,100±4,t=12.817,P＜0.05),but the LXA4+UA group was significantly decreased compared with the UA group (183±30,t=5.129,P＜0.05).Conclusion LXA4 inhibits UA induced ROS production in HUVECs.This mechanism might be through inhibiting p47phox trafficing from cytoplasm to cytomembrane,results in inhibiting the activation of NADPH oxidase.
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Objective To investigate the effects of lipoxin A4 ( LXA4 ) on inflammatory related factors induced by uric acid( UA) in human umbilical vein endothelial cells( HUVECs) . Methods HUVECs were treated with 5, 25, and 50 ng/ml LXA4 prior to exposure to 12 mg/dl UA. Tumor necrosis factor-alpha(TNF-α), interleukin ( IL)-1β, and IL-6 were analyzed with ELISA and realtime PCR. The phosphorylation levels of p38 mitogen-activated protein kinases( MAPK) and NF-κB/p65 were observed with Western blot. Results Stimulation of HUVECs with 12 mg/dl UA markedly increased TNF-α, IL-1β, and IL-6 production(P<0. 01). Pretreatment with LXA4 significantly inhibited UA-induced production of TNF-α, IL-1β, and IL-6 in a concentration dependent manner(P<0. 01). The mRNA expressions of TNF-α, IL-1β, and IL-6 in response to UA were also decreased by LXA4(P<0. 05). Western blot analysis showed that the phosphorylation levels of p38 MAPK and NF-κB/p65 were significantly raised by 12 mg/dl UA in HUVECs(P<0. 05), but attenuated significantly in the presence of 50 ng/ml LXA4. Conclusion LXA4 may inhibit the expressions of inflammatory related factors induced by UA via reducing p38 MAPK and NF-κB/p65 phosphorylation and play a role in anti-inflammation.
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Objective To investigate the prevalence of diabetes and prediabetes in northeast Sichuan area.Methods A total of 8 053 adult residents(over 18 years old)from the hospital who had underwent physical examination were recruited in the study.The concentrations of Fasting plasma glucose(FPG),glycated hemoglobin A1c(HbA1C)and blood lipids were measured.Diabetes and prediabetes were defined according to the 2013 American Diabetes Association criteria.Results The FPG and HbA1C concentra-tions increased with age.The overall prevalence of diabetes and prediabetes were estimated to be 8.5% and 39.6% in the adult pop-ulation in northeast sichuan area respectively.The prevalence of diabetes and prediabetes in men were significantly higher than those in women(10.7% vs.5.7%,41.5% vs.37.2%,P <0.05,respectively).Furthermore,the prevalence of diabetes increased with age,and the prevalence of prediabetes had a trend of first enhancement followed by a decline.Conclusion The prevalence of diabe-tes and prediabetes are relatively high in northeast Sichuan area.
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Objective To establish reference interval of hemoglobin A1c(HbA1c) determined in healthy adults in northeast Si-chuan .Methods Venous blood was pumped from 494 individuals without previously diagnosed diabetes and other critical illness . The HbA1c level ,blood routine examination ,blood lipids ,fasting plasma glucose ,liver and kidney function were measured . Reference value of HbA1c was determined by 95% confidence interval through the mean of HbA1c .Results The HbA1c level took on normal distribution in 494 healthy individuals ,and the reference interval was 4 .482% -6 .012% .There was no statistical signifi-cance of HbA1c level between different genders(t= -0 .905 ,P= 0 .366) .The levels of HbA1c in 20 - < 35 years old people , 35- <65 years old people and ≥65 years old people were(5 .109 ± 0 .150)% ,(5 .224 ± 0 .122)% ,(5 .444 ± 0 .125)% ,and the differences were statistically significant among different age group(P<0 .05) .The HbA1c level and age were positively correlated (r=0 .338 ,P<0 .01) .Conclusion It is necessary to establish appropriate reference intervals of HbA1c for different laboratories or areas .
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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.
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Objective To investigate the relationship of serum soluble CD40L with diabetic retinopathy(DR) in patients with type 2 diabetes mellitus(DM). Methods Serum soluble CD40L level was measured by ELISA in 30 diabetic patients without diabetic retinopathy (NDR), 29 patients with background diabetic retinopathy (NPDR), 31 patients with proliferative diabetic retinopathy (PDR), and 32 normal control(NC) subjects. Results The serum soluble CD40L levels were higher in NDR,NPDR and PDR groups [(4.55±3.66), (6.65±4.24), (8.31±5.23) μg/L,P<0.01] than in NC group[(2.01±1.35)μg/L]. Conclusions Sorluble CD40L level in serum is significantly increased in type 2 DM with diabetic retinopathy,which may be correlated with pathogenesis of DR
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0.05. The reference range of seum TIBC( ?1.96s) was 51.3-76.3?mol/L(70 cases).Conclusion The method was stable,simple, sensitive, rapid and accurate, and suitable for the routine manul and automatic analysis of total iron-binding capacity in serum.
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Experimental report is an important segment of the experimental teaching pro-cess.This article analyzes the current status and factors that influence the quality of experimental report,puts foreword some countermeasures on how to enhance the experimental teaching by im-proving the writing of experimental reports,in order to achieve the purpose of improving the quality of clinical experimental teaching of clinical biochemistry and laboratory.
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This article analyzes the current status and factors that influence the quality of practice teaching in medical laboratory,and presents the improvement of the status of practice teaching by basement construction,pre-education of clinical practice,standardized management,the cultivation of teachers,in order to achieve the purpose of improving the quality of clinical practice teaching
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Objective A enzymatic method for measurement of calcium in serum by using urea amidolyase was introduction. Methods Double reagent two-point method.TEA buffer(pH 8.0,200 mmol/L)was used as the buffer solution.The first reagent consisted of sodium bicarbonate 26.0 mmol/L,NADPH 0.4 mmol/L,2-oxoglutaric acid 8.0 mmol/L,urea amidolyase 115 U/L and GLDH 43.0 kU/L.The second reagent contained sodium bicarbonate 26.0 mmol/L and ATP 6.2 mmol/L in the buffer solution.Results The Linearity range was 0.5～5.0 mmol/L.The average rate of recovery was 102.7%.The average within-run and between-run CV were 2.13% and 2.69%.Camparison with OCPC(X1)，showed that Y1=1.02X1-0.021,r=0.981,and with Arsenazo Ⅲ(X2),Y2=0.99X2+0.03,r=0.978.No interference from as much as 300 μmol/L of bilirubin,4 g/L of hemoglobin,2.5 mmol/L of magnesium,2.0 mmol/L of ammonium ion and 8.0 mmol/L of triglyceride.Conclusion This method was simple,accurate and ripid,suitable for automatic analysis of calcium in serum.
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Objective To investigate the change of serum remnant like particle cholesterol (RLP C) and the association of RLP C with diabetic nephropathy (DN) in the patients with type 2 diabetes mellitus (DM). Methods Serum RLP C concentration was assayed by immunoseparation method. Plasma ? granule membrane glycoprotein 140 (GMP140) and nitric oxide (NO) levels were also measured. Urinary albumin excretion rate (UAER) was measured by radioimmunoassay, and the patients with type 2 DM were assigned to three groups according to UAER 〔35 without DN (D 1 group, UAER200 ?g/min)〕. Results Serum RLP C concentration in 96 patients with type 2 DM was significantly increased compared to that in 44 normal controls (NC) 〔(0.281?0.162)mmol/L vs (0.193?0.125)mmol/L, P