尿酸氧化酶与肌酐水解酶融合基因的克隆及其在大肠杆菌中的表达

目的 构建含尿酸氧化酶与肌酐水解酶融合基因的质粒并转化大肠杆菌,使其能同时分泌该两种酶分解尿酸及肌酐.方法 根据GenBank数据库已知的尿酸氧化酶基因设计引物,扩增到去掉终止密码子的尿酸氧化酶基因中,并使其连接到质粒pGM36e上构建成pGM36e-U,然后根据已知肌酐水解酶基因设计引物扩增肌酐水解酶基因,把其连接到质粒pGM36e-U中尿酸氧化酶基因后构建成重组质粒pGM36e-U/C,转化大肠杆菌,筛选鉴定重组大肠杆菌,并进行SDS-PAGE分析.将工程菌于特定浓度肌酐和尿酸的培养基中培养24h,取培养液测定肌酐和尿酸浓度.结果 重组质粒得到大小约1.68 kb基因片段.为尿酸氧化酶及肌酐水解酶之合.转化大肠杆菌后尿酸氧化酶及肌酐水解酶活性得到表达.肌酐分解率提高43.50％,尿酸分解率提高42.32％.结论 构建的含复合基因的重组质粒转化大肠杆菌后能表达尿酸氧化酶及肌酐水解酶活性.分解尿酸和肌酐.     

基于牛津郡社区卒中项目分型研究中风（中经络）急性期中医证型与检验指标相关性

目的 基于牛津郡社区卒中项目（Oxfordshire Community Stroke Project,OCSP）分型研究中风（中经络）急性期中医证型与检验指标的相关性。方法 收集225例中风（中经络）急性期患者,根据英国OCSP分型,中医辨证分型分为风痰阻络证、气虚血瘀证、风火上扰证、阴虚风动证。收集患者的既往史、生活习惯、检验指标资料,探讨中风（中经络）患者中医证型分布及OCSP分型与中医证型的关系、不同中医证型患者检测指标的变化。结果 中风（中经络）患者中医证型以风痰阻络证为主。不同性别患者中医证型分布差异无统计学意义（P>0.05）。不同OCSP分型患者4种中医证型分布存在差异（P<0.05）。不同证型患者血清肌酐水平比较,差异有统计学意义（P＜0.05）,不同证型患者尿酸、同型半胱氨酸、三酰甘油、低密度脂蛋白胆固醇、载脂蛋白A1水平比较,差异无统计学意义（P＞0.05）。结论 基于OCSP分型,中风（中经络）急性期中医证型与血清肌酐具有一定的相关性。     

在诊断糖尿病肾病中的价值

目的 评价随机尿白蛋白/肌酐比值(ACR)与血清胱抑素C(CysC)在诊断糖尿病肾病(DN)中的价值.方法 选取临床确诊的DN患者29例,未明确诊断为DN的糖尿病患者111例,正常对照组27例,使用免疫透射比浊法和酶法测定患者的随机ACR与血清CysC水平.结果 DN患者组中随机ACR与血清CysC水平均高于糖尿病未明...     

地高辛与中成药联用临床血清药物浓度监测

目的:;观察地高辛与中成药联用对其血清药物浓度及临床疗效的影响。方法:;取我院心血管内科符合充血性心衰诊断标准的住院患者30例,随机分为2组,每组15例,选择用丹参注射液与刺五加注射液分别与地高辛联用,观察2者对地高辛血清药物浓度及临床疗效的影响。结果:;2种中药注射液对地高辛的血清药物浓度没有影响,无1例不良反应发生,对心功能不全的病人的临床症状如胸闷等有所改善,这2种中药注射液分别与地高辛合用于心衰的治疗,对患者的肝、肾功能无明显影响。结论:;地高辛与中成药联用对其血清药物浓度影响不大。      

艾考糊精腹膜透析液对患者透析流出液中CA125水平的影响

目的 前瞻性研究使用艾考糊精透析液后患者透析流出液中CA125水平的变化,探讨其对腹膜间皮细胞的影响.方法 选取以2.5%葡萄糖透析液夜间留腹的持续性非卧床腹膜透析(CAPD)患者作为研究对象,随机分为艾考糊精组(选择7.5%艾考糊精透析液夜间留腹)和对照组(仍以2.5%葡萄糖透析液夜间留腹),总治疗期为4周,每2周随访1次(用药后第14、28天).观察和比较两组基线期和用药后第14、28天的夜间留腹净超滤量和腹膜肌酐清除率变化;测定两组基线期和用药后第28天的夜间留腹透析流出液中CA125水平,计算CA125呈现率.结果 54例CAPD患者入选,两组各27例.两组患者基线期各项指标比较差异均无统计学意义(P>0.05).艾考糊精组用药后第14、28天的净超滤量和腹膜肌酐清除率均较基线期明显提高,且显著高于相应时间点的对照组(P<0.05).艾考糊精组和对照组用药后第28天与基线期夜间留腹透析流出液CA125呈现率的差值分别为(1484.0±1571.6) U/h和(-222.1±1816.7)U/h,差异有统计学意义(P<0.01).结论 与2.5%葡萄糖透析液相比,7.5%艾考糊精透析液能显著提高患者透析流出液中CA125水平,提示腹膜间皮细胞生成数量增加.     

The role of renal proximal tubule P450 enzymes in chloroform-induced nephrotoxicity: Utility of renal specific P450 reductase knockout mouse models

The kidney is a primary target for numerous toxic compounds. Cytochrome P450 enzymes (P450) are responsible for the metabolic activation of various chemical compounds, and in the kidney are predominantly expressed in proximal tubules. The aim of this study was to test the hypothesis that renal proximal tubular P450s are critical for nephrotoxicity caused by chemicals such as chloroform. We developed two new mouse models, one having proximal tubule-specific deletion of the cytochrome P450 reductase (Cpr) gene (the enzyme required for all microsomal P450 activities), designated proximal tubule-Cpr-null (PTCN), and the other having proximal tubule-specific rescue of CPR activity with the global suppression of CPR activity in all extra-proximal tubular tissues, designated extra-proximal tubule-Cpr-low (XPT-CL). The PTCN, XPT-CL, Cpr-low (CL), and wild-type (WT) mice were treated with a single oral dose of chloroform at 200 mg/kg. Blood, liver and kidney samples were obtained at 24 h after the treatment. Renal toxicity was assessed by measuring BUN and creatinine levels, and by pathological examination. The blood and tissue levels of chloroform were determined. The severity of toxicity was less in PTCN and CL mice, compared with that of WT and XPT-CL mice. There were no significant differences in chloroform levels in the blood, liver, or kidney, between PTCN and WT mice, or between XPT-CL and CL mice. These findings indicate that local P450-dependent activities play an important role in the nephrotoxicity induced by chloroform. Our results also demonstrate the usefulness of these novel mouse models for studies of chemical-induced kidney toxicity.     

Factors associated with serum total homocysteine level in type 2 diabetes

Objectives The aim of this study was to investigate the determinants of serum total homocysteine level (tHcy) in patients with type 2 diabetes mellitus (DM) according to sex. Methods A total of 1,276 Japanese, diabetics (n = 280) with a control group of non-diabetics (n = 996), were enrolled into the study from 2003 to 2005. This cross-sectional study was conducted for all the subjects, using personal data regarding clinical characteristics and lifestyle. Multiple regression analysis was performed to analyze the association of tHcy with selected factors. Results In diabetic subjects, estimated glomerular filtration rate (eGFR) and serum creatinine levels (Cre), even those within the normal range, were strongly associated with tHcy after adjustment in both sexes; the standardized partial regression coefficient of eGFR for tHcy was −0.251, (p = 0.001) in diabetic men and −0.523, (p < 0.001) in diabetic women. Furthermore, the eGFR of the diabetics, except patients with nephropathy, also had significant association with tHcy in both sexes. Fasting plasma glucose levels and serum triglyceride levels were strongly associated with tHcy in diabetic men only. HbA1c was also associated with tHcy in diabetic men only, though not as significantly. Age and presence of hypertension were significantly associated with tHcy in women. Conclusions This study suggests that there are some differences in the factors associated with tHcy between diabetics and non-diabetics, and between the sexes. There is, therefore, circumstantial evidence that elevated tHcy should be evaluated clinically. Because tHcy was strongly associated with eGFR and Cre, even within the normal ranges, tHcy may have important implications regarding the microangiopathy of the kidney and atherosclerosis.     

Characterization of phthalate exposure among pregnant women assessed by repeat air and urine samples

BACKGROUND: Although urinary concentrations of phthalate metabolites are frequently used as biomarkers in epidemiologic studies, variability during pregnancy has not been characterized. METHODS: We measured phthalate metabolite concentrations in spot urine samples collected from 246 pregnant Dominican and African-American women. Twenty-eight women had repeat urine samples collected over a 6-week period. We also analyzed 48-hr personal air samples (n = 96 women) and repeated indoor air samples (n = 32 homes) for five phthalate diesters. Mixed-effects models were fit to evaluate reproducibility via intraclass correlation coefficients (ICC). We evaluated the sensitivity and specificity of using a single specimen versus repeat samples to classify a woman's exposure in the low or high category. RESULTS: Phthalates were detected in 85-100% of air and urine samples. ICCs for the unadjusted urinary metabolite concentrations ranged from 0.30 for mono-ethyl phthalate to 0.66 for monobenzyl phthalate. For indoor air, ICCs ranged from 0.48 [di-2-ethylhexyl phthalate (DEHP)] to 0.83 [butylbenzyl phthalate (BBzP)]. Air levels of phthalate diesters correlated with their respective urinary metabolite concentrations for BBzP (r = 0.71), di-isobutyl phthalate (r = 0.44), and diethyl phthalate (DEP; r = 0.39). In women sampled late in pregnancy, specific gravity appeared to be more effective than creatinine in adjusting for urine dilution. CONCLUSIONS: Urinary concentrations of DEP and DEHP metabolites in pregnant women showed lower reproducibility than metabolites for di-n-butyl phthalate and BBzP. A single indoor air sample may be sufficient to characterize phthalate exposure in the home, whereas urinary phthalate biomarkers should be sampled longitudinally during pregnancy to minimize exposure misclassification.