蛋白磷酸酶2A-Aα亚基基因5'侧翼区多态性在广东汉族人群中的分布

目的:探讨蛋白磷酸酶2A-Aα亚基基因PPP2R1A启动子区的遗传多态性在中国南方汉族人群中的分布特征.方法:随机选取部分广东汉族人群,获取全血基因组DNA.PCR扩增获得PPP2R1A基因5'-侧翼区的目的片段产物直接再测序,筛查并确证潜在的基因多态性位点,采用HaploView软件进行该人群多态性等位基因分布频率的分析、并与HapMap结果进行分布特征比较.结果:PCR扩增和成功测序63例健康人(126条染色体)PPP2R1A基因-1 844～+201 nt的目的片段,序列比对发现该人群中6个已知多态性位点及其人群最小等位基因频率分别为-1 039 G>T(+Ins)(rs10414793,但其中T等位基因型含有29个碱基片段的插入)(8.73%)、-568 G>A(rs1864007)(25.40%)、-241 -/G(rs11453459)(26.90%)、+87 T>C(rs13344984)(7.94%)、+107 -/C(rs3833207)(30.16%)和+108 A>G(rs17554825)(7.94%);且其中2个位点在该人群中的分布与HapMap公布的国外人群数据存在不同(P<0.05);同时,该人群中还发现-512 G>A(2.38%)的潜在新多态性位点.结论:筛查PPPwR1A基因5'-侧翼区在中国广东汉族人群中多态性位点的等位基因分布,为进一步开展多态性功能性分析提供基础.     

雄激素受体基因E211G》A多态性与汉族女性青春期后痤疮的关联性分析

目的:探讨雄激素受体(AR)基因E211 G>A单核苷酸多态性与汉族女性青春期后痤疮的关联性,为汉族女性青春期后痤疮发病遗传学研究奠定基础.方法:将女性青春期后痤疮患者79例和正常女性志愿者80例分为女性青春期后痤疮组和女性正常对照组,采用PCR技术扩增出AR-E211 G>A基因多态位点的片段,用限制性内切酶StuⅠ进行酶切,产物在2%琼脂糖凝胶上电泳,确定G>A基因的3种基因型,即GG、GA、AA,观察两组基因型频数和等位基因型频数分布的差异.结果:女性青春期后痤疮组GA+AA基因型频数低于对照组(P=0.028,OR=0.937,95% CI=0.885～0.992).女性青春期后痤疮组A等位基因频数低于对照组(P=0.029,OR=0.968,95% CI=0.941～0.996).结论:AR-E211 A等位基因的存在使青春期后女性患痤疮的风险性明显降低.     

人甘露糖结合凝集素相关丝氨酸蛋白激酶-2C端基因的克隆与鉴定

目的:获得人甘露糖结合凝集素相关丝氨酸蛋白激酶-2(MASP-2)C端编码基因,并表达人MASP-2 C端片段,为制备单克隆抗体及应用于临床相关疾病的检测奠定分子学基础.方法:采用RT-PCR技术从人胎肝组织总RNA中扩增人MASP-2 C端的cDNA,克隆入pGEX-6p-2表达载体,酶切图谱分析和序列分析鉴定.结果:获得编码人MASP-2 C端片段的cDNA,并且与pGEX-6p-2载体连接,转化大肠杆菌XL1-blue,成功构建人MASP-2 C端片段cDNA的重组克隆.重组质粒酶切图谱分析和DNA测序分析,人MASP-2 C端片段cDNA的重组克隆与GenBank中人MASP-2 C端的cDNA序列一致.结论:成功克隆了人MASP-2 C端片段cDNA,并在大肠杆菌中表达人MASP-2 C端多肽片段.     

Insertion/deletion polymorphism in the angiotensin-converting enzyme gene associated with macroangiopathy and blood pressure in patients with non-insulin-dependent diabetes mellitus

In the search for new risk factors for diabetic macroangiopathy the insertion/deletion (I/D) polymorphism in the angiotensin-converting enzyme gene was studied in 237 consecutive patients (125 men and 112 women) with non-insulin-dependent diabetes. The female population showed an excess of ischemic electrocardiographic changes or definite myocardial infarctions in the patients homozygous for the deletion [D/D; odds ratio (OR) 2.8; 95% confidence interval (CI) 1.4-5.3] and in the insertion/deletion heterozygotes (I/D; OR 1.8; CI 1.1-3.1) compared with the patients homozygous for the insertion (I/I). In the total series coronary heart disease, cerebrovascular disease, and claudication were more often observed in the patients with I/D (OR 1.5; CI 1.0-2.2) or the D/D genotype patients (OR 1.7; CI 1.1-2.6) than in those with the genotype I/I. The systolic blood pressure was lower in patients with genotype I/I (138 +/- 19 mmHg) than in those with the genotype I/D (149 +/- 22 mmHg) or D/D (150 +/- 21 mmHg; P < 0.02). The prevalence of hypertension and the median urinary albumin excretion rate also tended to be lowest in the I/I genotype patients. Multiple logistic analysis revealed that in women the angiotensin-converting enzyme D/D genotype is independently associated with coronary heart disease. Our findings suggest that variation at the angiotensin-converting enzyme gene locus is one of the factors involved in the predisposition of diabetic patients to the development of arterial disease and hypertension.     

The prevalence of type 2 diabetes and gestational diabetes mellitus in an inner city multi-ethnic population

Zeeburg', a multiethnic town borough in the Amsterdam-East region, has one of the city's highest rates of immigrants. In the total population of 19,825 Surinam (mainly Creole), Turkish, Moroccan, and Dutch adults the prevalence of known type 2 diabetes in 1994 and of gestational diabetes mellitus (GDM) between January 1992 and January 1997 was investigated. Based on World Health Organization (WHO) criteria of 1985, the age-standardized prevalence of type 2 diabetes was similar in men (6.4%; 95% confidence interval [CI]: 5.6-7.2) and women (6.4%: 95% CI: 5.8-7.0) for all ethnic groups combined. However, the age- and sex-standardized prevalence of type 2 diabetes was significantly greater in the non-Dutch inhabitants than in the Dutch inhabitants (17.3% [95% CI: 12.9-21.6] in Surinam inhabitants, 10.9% [95% CI: 9.7-12.2] in Turkish inhabitants, 12.4% [95% CI: 9.7-15.0] in Moroccan inhabitants, and 3.6% [95% CI: 3.2-3.9] in Dutch inhabitants). The odds ratios for type 2 diabetes for the separate immigrant groups relative to the Dutch group were 5.88 (95% CI: 4.54-7.69) for Surinam inhabitants, 4.00 (95% CI: 2.86-5.55) for Turkish inhabitants, and 4.17 (95% CI: 3.03-5.55) for Moroccan inhabitants. GDM was present in 2.59% of women of non-Dutch origin compared with 0.62% of women of Dutch origin. A significant positive association was found between the non-Dutch origin and the occurrence of GDM (chi2 = 6.7; p < 0.01). The study highlights a high prevalence of known type 2 diabetes and GDM in the immigrant inhabitants and emphasizes that appropriate interventions are necessarily with implications for health targets and capitation based budgets.     

Comparison of granulocyte function in diabetes mellitus type 1 and type 2

The phagocytosis rate of polymorphonuclear leucocytes was measured by flow-cytometry. Vital bacteria were incubated in whole blood. 111 blood samples were measured, 54 in diabetic patients (14 type 1 and 40 type 2), the rest of 57 samples in healthy controls. Results showed firstly, that a decompensation in glucose metabolism in diabetic patients correlated with a decrease in phagocytosis. The HbA1 level was more closely correlated than the glucose level. The second result was, that despite a similar grade of decompensation in type 1 and type 2 diabetic patients, the phagocytosis was significantly lower in type 1 diabetes. No correlation was found concerning age and sex. These findings show, that the impact on granulocytic function in diabetes is of multifactorial origin, not only a shorter or longer elevation of the serum glucose level can explain it solely.     

The association between a family history of type 2 diabetes and coronary artery disease in a type 1 diabetes population

The plasma protein beta2-glycoprotein I (beta2-GPI) is a major target of autoantibodies in patients with the antiphospholipid syndrome. To understand the physiological function of beta2-GPI and its potential role in the pathophysiology of the antiphospholipid syndrome, the binding of beta2-GPI to phospholipid membranes was characterized. The interaction of beta2-GPI with unilamellar vesicles containing varying amounts of acidic phospholipids with phosphatidylcholine (PC) was measured at equilibrium via relative light scattering. Analysis of binding isotherms gave apparent Kd values ranging from approximately 5.0 to 0.5 microM over a range of 5-20 mol % anionic phospholipid. Inhibition of binding by increasing ionic strength and Ca2+ ions suggests that binding is primarily electrostatic. These data indicate that beta2-GPI binding to membranes with physiological anionic phospholipid content is relatively weak in comparison to plasma coagulation proteins, suggesting that beta2-GPI does not function as a physiological anticoagulant based on its phospholipid-binding properties.     

Combination insulin and sulfonylurea therapy in insulin-requiring type 2 diabetes mellitus

OBJECTIVES: To identify ethical dilemmas experienced by occupational and physical therapists working in the UK National Health Service (NHS). To compare ethical contexts, themes and principles across the two groups. DESIGN: A structured questionnaire was circulated to the managers of occupational and physical therapy services in England and Wales. SUBJECTS: The questionnaires were given to 238 occupational and 249 physical therapists who conformed to set criteria. RESULTS: Ethical dilemmas experienced during the previous six months were reported by 118 occupational and 107 physical therapists. The two groups were similar in age, grade, and years of experience. Fifty of the occupational therapy dilemmas occurred in mental health settings but no equivalent setting emerged for physical therapy. Different ethical themes emerged between the two groups, with the most common in occupational therapy being difficult/dangerous behaviour in patients and unprofessional staff behaviour, and for physical therapists resource limitations and treatment effectiveness. No differences were found in the ethical principles used. CONCLUSION: The ethical dilemmas reported by the therapists were primarily concerned with health care ethics, rather than the more dramatic ethics reported in much of the biomedical ethics literature. Differences were found between the two professional groups when ethical contexts and themes were compared but not when ethical principles were compared. This suggests that educators and researchers need to be aware of work settings and the interdisciplinary nature of employment as well as ethical principles held by individual therapists.     

Diabetic retinopathy, promoter (4G/5G) polymorphism of PAI-1 gene, and PAI-1 activity in Pima Indians with type 2 diabetes

OBJECTIVE: To examine the relationship between plasma plasminogen activator inhibitor 1 (PAI-1) activity and PAI-1 gene (4G/5G) polymorphism and diabetic retinopathy in Pima Indians with type 2 diabetes. RESEARCH DESIGN AND METHODS: We studied 171 Pima Indians with type 2 diabetes between the ages of 30-70 years in a population-based epidemiological survey. Plasma PAI-1 activity was measured by a spectrophotometric assay and PAI-1 4G/5G promoter genotype by the polymerase chain reaction (PCR) using allele-specific primers. Retinopathy was assessed by ophthalmoscopy after pupillary dilation and classified as any retinopathy or as nonproliferative and proliferative. RESULTS: Retinopathy was present in 70 (41%) subjects, and 4 (2.3%) subjects had proliferative retinopathy. Plasma PAI-1 activity was not significantly different among subjects with and without retinopathy (17.1 +/- vs. 19.7 +/- 9.1 arbitrary units (AU)/ml, P = 0.09). PAI-1 activity was negatively correlated with duration of diabetes (rs = -0.18, P = 0.02). In a logistic regression analysis controlled for age, sex, BMI, and duration of diabetes, any retinopathy was significantly associated with fasting plasma glucose concentrations (P < 0.05), 2-h postload glucose (P = 0.02), and HbA1c (P = 0.008), but not with PAI-1 activity (P = 0.48). The prevalence of retinopathy in the three genotype groups differed significantly (4G/4G, 4G/5G, and 5G/5G were 44, 49, and 24%, respectively; chi 2 = 8.22, df = 2, P = 0.016) and remained significant after controlling for age, sex, BMI, duration of diabetes, glycated hemoglobin, and urine albumin-to-creatine ratio in a logistic regression analysis. The odds ratios for retinopathy in subjects with 4G/4G and 4G/5G, compared with the 5G/5G genotype, were 2.0 and 3.1, respectively. CONCLUSIONS: Although diabetic retinopathy in Pima Indians with type 2 diabetes is not associated with PAI-1 activity, subjects with the 4G/4G and 4G/5G genotype had a higher prevalence of retinopathy compared with 5G/5G PAI-1genotype. These preliminary findings indicate that in Pima Indians with type 2 diabetes, presence of the 4G allele of the PAI-1 gene was associated with a higher risk of diabetic retinopathy.     

Effect of metformin on bile salt circulation and intestinal motility in type 2 diabetes mellitus

Theophylline anhydrate microcapsules with different amounts of MA/MMA copolymer (Eudragit L) were prepared by the solvent evaporation method. Qualitative as well as quantitative investigation of the drug-polymer interaction by Fourier transform infrared (FTIR) spectroscopy with a curve fitting program was undertaken. The release mechanisms of theophylline in pH 1.2 and pH 6.8 media were also studied to elucidate the effect of drug-polymer interaction on the release characteristics of microcapsules. Direct evidence for a hydrogen bonding interaction between theophylline and Eudragit L in microcapsules was obtained. Moreover, the fraction of hydrogen bonded theophylline increased with the increase of Eudragit L. The dissolution of theophylline from microcapsules exhibited an enteric-coated release property. The drug release mechanism was found to fit the Higuchi matrix model in the simulated gastric acid condition, but drug release was much more rapid in the pH 6.8 buffer solution. The drug release rate decreased as the composition of theophylline increased, and it was proportional to the fraction of hydrogen bonded theophylline. These results suggest that the increased fraction of hydrogen bonded theophylline in microencapsulation might improve the mixing and dispersibility of theophylline in the Eudragit L matrix, thus resulting in the increase of the release rate of theophylline from microcapsules.     

Relationships between serum lipids, platelet membrane fatty acid composition and platelet aggregation in type 2 diabetes mellitus

In order to gain further insight into the mechanism of platelet dysfunction frequently reported in diabetes we investigated circulating fatty acids, lipid composition of platelet membrane and platelet function in Type 2 diabetic patients. In these subjects, percentages of C16 : 1n-7 and C18 : 1n-9 in serum phospholipid fraction and of C16 : 1n-7 in serum cholesterol ester fraction were decreased. Moreover, the content of C20 : 4n-6 in serum cholesterol esters was altered in Type 2 diabetic subjects: C18 : 0 and C20 : 3n-6 were increased but C20 : 4n-6 content was similar to controls. Aggregation in vitro did not differ from controls but aggregation in vivo was increased in Type 2 diabetic subjects. No correlation was observed between metabolic parameters -i.e., HbA1, blood glucose, serum triglycerides and total cholesterol, circulating fatty acids and fatty acid content of platelet membrane. A negative linear correlation was found between aggregation in vivo and C20 : 4n-6 content of platelet membrane. Moreover, a U shaped relationship was observed between platelet aggregation in vitro and C20 : 4n-6 content of platelet membrane suggesting that C20 : 4n-6 level should be tightly controlled otherwise platelet hyperreactivity may occur. These results indicate that despite a normal mean C20 : 4n-6 content in the platelet membrane, regulation of C20 : 4n-6 metabolism is less strictly controlled in Type 2 diabetes mellitus and confirm the importance of arachidonic acid platelet content in the regulation of platelet aggregability.     

Pharmacodynamics of insulin Lispro in 2 patients with type II diabetes mellitus

We studied the effects of the new rapid acting human insulin analogue (Lys(B28), Pro(B29) insulin), insulin Lispro (Lispro) on metabolic control and insulin receptor binding in type II diabetes mellitus. We investigated 2 patients: Patient 1 was obese, clearly insulin-resistant, injected high doses of insulin (3-4 IU/kg body weight), and had insufficient diabetes control. Patient 2 was of normal body weight, injected normal insulin doses (0.7-0.8 IU/kg body weight), and had good diabetes control. Patient 1 showed a considerable improvement of insulin binding after receiving Lispro (26,700 vs. 5,600 receptors/monocyte; Kd 560 vs. 1,500 pM). Concommitantly, a decrease of serum glucose and insulin dose was observed, reflecting a higher insulin sensitivity during Lispro treatment. In patient 2 injected with Lispro the time course of serum glucose, serum insulin, and insulin binding after an oral meal was comparable to values obtained in healthy controls. We conclude that the quick and pulsatile pharmacokinetic profile of the insulin analogue Lispro may improve glycemia, insulin receptor binding, and insulin resistance in type II diabetes.