冠心病多位点β纤维蛋白原基因多态性检测的临床意义

目的研究纤维蛋白原(Fg)Bβ基因多个位点的多态性与血浆水平及冠心病的关系。方法运用聚合酶链反应-限制性片段长度多态性分析技术及核苷酸序列测定鉴定FgBβ基因中7个可能与血栓相关的多态性位点:-β148C/T-、249 C/T3、45 C/T、-455G/A-、854 G/A、1689T/G及Bc1 I G/A;比浊法测定血浆纤维蛋白原水平。结果与正常对照组相比,冠心病组血浆Fg显著升高(P<0.05);有-148C/T-、455G/A、-854G/A或Bc1 I G/A基因多态性变异组,其血浆Fg水平高于无变异组(P<0.05),其中,同时携带A-455、A-854者增高更为显著(P<0.01)。冠心病组A-455、T-148基因型频率(0.334)显著高于高血压组(0.196)和正常对照组(0.195),多态性位点G-455、C-148或A-455、T-148分别紧密连锁,符合率超过97%;Logistic回归分析发现,携带FgB-β148T-、455A基因的高血压患者,患冠心病的危险性是非携带者的1.654倍(P=0.01,95%CI:1.207-2.267)。结论FgBβ基因多态性与血浆Fg水平及缺血性心脏病发生的危险性相关;选择性FgBβ基因多态性位点的检测有助于临床上冠心病易患人群的筛查。     

纤维蛋白原基因多态性与缺血性心脑血管病的关系

目的：调查健康人、心肌梗死患者及脑梗死患者的纤维蛋白原（Fg)β-455G/A、-148C/T、448G/A基因多态性频率分布、Fg分子反应性及与血浆Fg水平的关系。方法：用限制性片段长度多态性分析基因频率分布，用计算机辅助的纤维蛋白单体聚合反应分析方法和Clauss法分析血浆Fg水平。结果：等位基因－455A、－148T和448A在正常人中的频率分别是0．185，0．194及0．192；在心肌梗死患者中的频率分别是0．295，0．318及0．307；在脑梗死患者中的频率分别是0．177，0．193及0．182。心肌梗死患者中－455A、－148T和448A的频率比健康人明显升高。3个多态性位点－455G、－148C和448G或－455A、－148T和448A分别紧密连锁，符合率超过98％。心脑血管病患者的Fg功能明显增高且与Fg水平相关。3个多态性位点不同基因型组血浆Fg水平差异无显著性。结论：3对等位基因紧密连锁不平衡，不同基因型组血浆Fg水平差异无显著性，心肌梗死患者中－455A、－148T和448A的频率比健康人明显升高。提示Fgβ-455G/A、-148C/T和448G／A三种基因多态性与血浆Fg水平无关，而与心肌梗死的发病相关。心脑血管病患者不仅Fg功能明显增高，且与Fg水平相关。     

纤维蛋白原紧密连锁不平衡基因位点BβG/A-455、C/T-148、G/A+448与缺血性脑血管病相关性分析

目的 分析纤维蛋白原(FIB)3个紧密连锁不平衡基因位点BβG/A-455、C/T-148、G/A+448基因多态性及血浆FIB水平与缺血性脑血管病的相关性.方法 以我院2010年2月至2012年2月收治的缺血性脑血管病患者89例为病例组,以同期在我院健康体检的50例为对照组,检测两组血浆FIB水平,使用多聚酶链式反应限制性片段长度多态性(PCR-RFLP)技术测定BβG/A-455、C/T-148、G/A+448基因片段.结果 缺血性脑血管病患者中,FIB基因型分布、BβA-455、T-148、A+448等位基因频率与对照组比较差异有统计学意义(P＜0.05).两组血浆FIB水平比较差异有统计学意义(P＜0.01),病例组高于对照组.A、T等位基因携带者与非携带者血浆FIB水平比较差异有统计学意义(缺血性脑血管病患者组,P＜0.01;对照组,P＜0.05),携带者高于非携带者.结论 FIB 3个紧密连锁不平衡基因位点BβG/A-455、C/T-148、G/A+448基因多态性及血浆FIB水平与缺血性脑血管病呈明显的正相关,BβG/A-455、C/T-148、G/A+448基因多态性可能是缺血性脑血管病的遗传易感因素.     

β—纤维蛋白原—455G／A基因多态性对缺血性脑卒中患者血浆纤维蛋白原水平的影响

为了分析β-纤维蛋白原-455G/A(β-Fg-455G/A)基因多态性与环境因素对血浆Fg水平及缺血性脑卒中发病的影响,应用聚合酶链反应(PCR)及限制性内切酶分析的方法,分析了104例缺血性脑卒中患者及156例健康人的β-Fg-455G/A基因多态现象,用比浊法测定血浆Fg水平.研究结果显示,病例组血浆Fg水平明显高于对照组(P＜0.01).无论男、女患者与对照相比,A等位基因携带者血浆Fg水平均比同组GG基因型者明显升高(P＜0.05).对照组内只有A等位基因携带者随年龄增长血浆Fg水平有明显升高(P＜0.05).病例组男性按吸烟及基因型情况分组,在GA基因型中,吸烟组血浆Fg水平高于不吸烟与戒烟组(P＜0.05);GG基因型组吸烟与否对Fg水平无显著影响(P＞0.05).病例组与对照组A等位基因频率分布无差异.结论提示,A等位基因携带者血浆Fg水平升高,并随年龄、吸烟而更加显著,提示β-Fg-455 A等位基因携带者血浆Fg水平更易受环境因素影响而升高,故此基因多态性可用于易感人群的检测,对缺血性脑卒中早期防治有一定意义.     

缺血性脑血管疾病发病的遗传学因素:南宁地区人群β纤维蛋白原基因-455G/A多态性与纤维蛋白原的观察

目的:调查广西南宁地区人群是否存在β纤维蛋白原基因-455G/A多态性及其与纤维蛋白原表达的关系,并与其他地区比较。方法:调查于2003-11/2004-05在广西医科大学第一附属医院进行,应用聚合酶联反应-限制性片断长度多态性(PCR-RFLP)技术检测南宁地区123份DNA的β纤维蛋白原基因-455G/A多态性,用PT衍生法测定全部123份血浆纤维蛋白原水平;检索国内及欧洲部分文献中β纤维蛋白原基因-455G/A基因型频率后与之比较。结果:检出β纤维蛋白原基因-455G/A多态性,GG型为常见基因型,G/A等位基因频率为0.789/0.211。基因频率与欧洲部分国家相近(P>0.05),但在国内存在差异。A等位基因组纤维蛋白原量为(3.4585±0.7231)g/L,G等位基因组纤维蛋白原量为(3.5058±0.7171)g/L,两等位基因组纤维蛋白原量比较差异无显著性意义(P=0.7251)。结论:南宁地区多人群存在β纤维蛋白原基因-455G/A多态性,该基因多态性可能存在一定的地域性差异。G/A等位基因对血浆纤维蛋白原水平的影响不大。     

纤维蛋白原β-455G/A、-148C/T、448G/A基因多态性与血浆纤维蛋白原水平的关系

目的　调查 15 6名广东汉族健康人纤维蛋白原 (Fg) β - 45 5G/A、- 148C/T、44 8G/A基因多态性频率分布、连锁不平衡关系及与血浆Fg水平的关系。方法　用限制性片段长度多态性 (RFLP)分析方法和比浊法检测血浆Fg水平。结果　等位基因A-4 55、T-14 8和A4 48的频率分别是 0 .2 76 ,0 .2 85及 0 .2 72。 15 6人中 3个多态性位点G-4 55、C-14 8和G4 48或A-4 55、T-14 8和A4 48分别紧密连锁 ,符合率超过97%。 3个多态性位点携带及不携带突变基因两组血浆Fg水平分别是 β - 45 5 :(3 .13± 0 .74)g/L和(2 89± 0 .5 7)g/L ;β - 148:(3 .12± 0 .73)g/L和 (2 .89± 0 .5 8)g/L ;β 44 8:(3 .13± 0 .74)g/L和 (2 89±0 5 7)g/L。上述 3个位点携带突变基因组血浆Fg水平均比野生型组明显升高 (P <0 .0 5 )。结论　 3对等位基因紧密连锁不平衡 ,携带突变基因者较不携带者血浆Fg水平高 ,提示Fgβ - 45 5G/A、- 148C/T、44 8G/A 3种基因多态性均与血浆Fg水平关联     

β-纤维蛋白原-455 G/A基因多态性与脑梗死的关系

目的:探讨β-纤维蛋白原-455 G/A(-βFg-455G/A)基因多态性与中国东北地区汉族脑梗死发病(CI)的关系。方法:应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法,检测脑梗死组(CI组)72例和对照组69例-βFg-455G/A基因的多态性,并测定其血浆纤维蛋白原(Fg)的含量。结果:经χ2检验,各基因型频率和等位基因频率在两组间均无显著性差异(P>0.05);CI组血浆Fg水平(2.70±1.05 g/L)和对照组(2.62±0.51 g/L)比较差异无显著性意义(P>0.05)。结论:本研究未发现-βFg-455G/A基因多态性与脑梗死之间存在相关关系。     

β-纤维蛋白原-455 G/A基因多态性的研究进展

多篇流行病学调查报道血浆纤维蛋白原(Fg)水平与心脑血管疾病、静脉血栓形成以及外周动脉性疾病强相关,而β-纤维蛋白原-455G/A基因多态性可导致血浆纤维蛋白原水平增高,从而引起高凝状态,致使血栓发病率升高。本文综述了近年来关于β-纤维蛋白原-455G/A基因多态性的研究进展。     

儿童单纯性肥胖与纤维蛋白原Bβ-148C/T、Bβ448G/A基因多态性关系的研究

目的 研究纤维蛋白原Bβ-148C/T、Bβ448G/A 基因多态性与儿童单纯性肥胖的相关性,为防治儿童单纯性肥胖提供理论依据.方法 抽取空腹静脉血采用聚合酶链反应限制性酶切方法对纤维蛋白原Bβ-148C/T、Bβ448G/A位点的基因型进行测定.结果 发现Bβ-148C/T基因型的分布在单纯性肥胖组和正常体重组差异有统计学意义(CC 51/67,CT 47/37,TT 8/2,基因型分布P=0.03);儿童单纯性肥胖T等位基因频率明显高于健康对照组(C 149/171,T 63/41,等位基因频率P=0.02),而B13448G/A基因型及等位基因频率的分布在单纯性肥胖组和正常体重组差异无统计学意义(GG 61/69,AG41/32,AA4/5,G 163/70,A49/42,均为P0.05).结论 纤维蛋白原Bβ-148 C/T基因多态性与儿童单纯性肥胖有相关性.而Bβ448G/A基因多态性与儿童单纯性肥胖不相关.     

纤维蛋白原Bβ-148C/T基因多态性与脑卒中的关系

目的:探讨纤维蛋白原(Fg)Bβ基因启动子区的多态性位点-148C/T对血浆Fg水平的影响及与脑卒中的关系。方法:利用聚合酶链反应-限制片段长度多态性(PCR-RELP)方法分析基因多态性,比浊法测定血浆Fg水平。对脑卒中组和对照组进行检测和分析。结果:发现-148TT等位基因频率在脑梗死组、脑出血组分别为0.2444、0.2500明显高于对照组0.1750(P<0.05);且-148CT/TT的脑梗死组、脑出血组的Fg水平分别为(3.23±1.09)g/L、(3.36±0.58)g/L,也明显高于对照组(2.85±0.76)g/L。结论:FgBβ基因启动子区-148C/T多态性与脑卒中的发生相关,并可影响血浆Fg水平,可能是脑卒中发病的遗传学危险因素。     

Enhanced atherosclerosis and kidney dysfunction in eNOS(-/-)Apoe(-/-) mice are ameliorated by enalapril treatment

Hypertension and atherosclerosis are each important causes of morbidity and mortality in the developed world. We have investigated the interaction between these conditions by breeding mice that are atherosclerotic due to lack of apolipoprotein (apo) E with mice that are hypertensive due to lack of endothelial nitric oxide synthase (eNOS). The doubly deficient mice (nnee) have higher blood pressure (BP) and increased atherosclerotic lesion size but no change in plasma lipoprotein profiles compared with normotensive but atherosclerotic (NNee) mice. The nnee mice also develop kidney damage, evidenced by increased plasma creatinine, decreased kidney weight/body weight ratio, and glomerular lipid deposition and calcification. Enalapril treatment abolishes the deleterious effects of eNOS deficiency on BP, atherosclerosis, and kidney dysfunction in nnee mice. In striking contrast, a genetic lack of inducible NOS, which does not affect BP, has no effect on the development of atherosclerotic lesions in Apoe(-/-) mice. We also observed a positive relationship between BP and size of atherosclerotic lesions These results suggest that the atherogenic effects of eNOS deficiency can be partially explained by an increase in BP and reemphasize the importance of controlling hypertension in preventing atherosclerosis.     

Hydronephrosis associated with antiurothelial and antinuclear autoantibodies in BALB/c-Fcgr2b-/-Pdcd1-/- mice

Because most autoimmune diseases are polygenic, analysis of the synergistic involvement of various immune regulators is essential for a complete understanding of the molecular pathology of these diseases. We report the regulation of autoimmune diseases by epistatic effects of two immunoinhibitory receptors, low affinity type IIb Fc receptor for IgG (FcgammaRIIB) and programmed cell death 1 (PD-1). Approximately one third of the BALB/c-Fcgr2b(-/-)Pdcd1(-/-) mice developed autoimmune hydronephrosis, which is not observed in either BALB/c-Fcgr2b(-/-) or BALB/c-Pdcd1(-/-) mice. Hydronephrotic mice produced autoantibodies (autoAbs) against urothelial antigens, including uroplakin IIIa, and these antibodies were deposited on the urothelial cells of the urinary bladder. In addition, approximately 15% of the BALB/c-Fcgr2b(-/-)Pdcd1(-/-) mice produced antinuclear autoAbs. In contrast, the frequency of the autoimmune cardiomyopathy and the production of anti-parietal cell autoAb, which were observed in BALB/c-Pdcd1(-/-) mice, were not affected by the additional FcgammaRIIB deficiency. These observations suggest cross talk between two immunoinhibitory receptors, FcgammaRIIB and PD-1, on the regulation of autoimmune diseases.     

Mechanical, thermal and formalin-induced nociception is differentially altered in 5-HT1A-/-, 5-HT1B-/-, 5-HT2A-/-, 5-HT3A-/- and 5-HTT-/- knock-out male mice

Extensive studies in rodents suggest that serotonin (5-HT) modulates nociceptive responses through the stimulation of several receptor types. However, it remains to demonstrate that these receptors participate in the control of nociception under physiological conditions. Pain behaviors of mutants which do not express 5-HT_1A, 5-HT_1B, 5-HT_2A or 5-HT_3A receptors, or lacking the 5-HT transporter, compared to paired wild-type mice of the same genetic background, were examined using validated tests based on different sensory modalities. Mechanical (von Frey filaments, tail pressure, tail clip tests), thermal (radiant heat, 46 °C water bath, hot-plate test) and formalin-induced nociception were determined in 2- to 3-month-old males. 5-HT_1A knock-out mice differed from wild-types by higher thermal sensitivity (hot-plate test only), and 5-HT_1B knock-out mice by higher thermal and formalin sensitivity. Both 5-HT_2A and 5-HT_3A knock-out mice differed from wild-types by a dramatic decrease in the formalin-induced nociceptive responses for phase II (16–45 min after injection/inflammatory phase). In contrast, neither mechanical, thermal nor formalin-induced nociception differed between mutants lacking the 5-HT transporter and paired wild-type mice. Although differences in spontaneous locomotor activity in 5-HT_1B−/− (increase) and 5-HT_3A−/− (decrease) knock-out mice versus paired wild-types might have confounded differences in nociception, acute 5-HT receptor blockade by selective antagonists was found to replicate in wild-type mice the effects on pain behavior, but not on locomotor activity, of the respective gene knock-out in mutants. These results support the conclusion that the complex control of pain mechanisms by 5-HT, acting at multiple receptors, is physiologically relevant in mice.     

Acoustic quantification of /i/-/I/ overlap in children 21 to 33 months

This longitudinal investigation examined the temporal and spectral characteristics of the high front vowels /i/ and /I/ as produced by nine monolingual US English children from 21–33 months. Vowel overlap was quantified in two-dimensional (F1, F2) and three-dimensional (F1, F2, duration) space using Spectral Overlap Assessment Measure (SOAM). These findings were compared with the results from Support Vector Machine (SVM) vowel classification, vowel duration ratios, and measures of effect size, to determine whether a spectral/temporal trading effect existed in the early vowel productions of young children. Children between the ages of 21 and 33 months are highly variable in the way they use spectral and temporal parameters to distinguish between these two adjacent vowels. However, findings pointed to the existence of a spectral/temporal trading effect when spectral overlap values are relatively high (>60%) at 21 and 24 months of age.     

瑞舒伐他汀钙对apoE-/-小鼠动脉粥样硬化血清抵抗素的影响

目的研究瑞舒伐他汀钙对小鼠动脉粥样硬化血清抵抗素的影响。方法将高脂喂养的雄性apoE-/-小鼠24只,随机分为模型组(n=12)、瑞舒伐他汀钙组(n=12,加用瑞舒伐他汀10mg.kg-1.d-1蒸馏水稀释后灌胃)。雄性C57BL/6J野生型小鼠12只作为正常对照组。12周后,处死小鼠,收集血清,测定血脂及抵抗素的含量。取主动脉根部做病理切片,HE染色后,检测校正斑块面积。结果瑞舒伐他汀钙组较模型组血中TC、TG、LDL-C浓度有显著下降(P<0.01)。瑞舒伐他汀钙组动脉粥样硬化病变的校正斑块面积显著低于模型组(P<0.01)。模型组与正常对照组比较,血清抵抗素浓度明显升高(P<0.01)。瑞舒伐他汀钙组较模型组血清抵抗素浓度显著降低(P<0.01)。结论瑞舒伐他汀钙可有效地降低动脉粥样硬化小鼠的血清抵抗素含量,显著抑制炎症反应,抑制粥样斑块进展,从而减轻动脉粥样硬化,此可能为他汀类药物抗动脉粥样硬化的机制之一。     

Genetic modifiers of systemic lupus erythematosus in FcgammaRIIB(-/-) mice

FcgammaRIIB is a potent lupus susceptibility gene as demonstrated by the observation that mice deficient in this molecule develop spontaneous antinuclear antibodies (ANA) and fatal glomerulonephritis when on the C57BL/6 background. To determine the mechanisms underlying the epistasis displayed by this gene we have constructed hybrids between FcgammaRIIB(-/-) and the systemic lupus erythematosus (SLE) modifiers yaa and lpr and the susceptibility locus Sle1. Sle1 and B6.RIIB(-/-) are both physically and functionally coupled; compound heterozygotes of Sle1 and B6.RIIB(-/-) develop significant disease, while single heterozygotes display no evidence of autoimmunity or disease, indicating that these genes lie on the same genetic pathway resulting in the loss of tolerance to nuclear antigens. However, the generation of ANA in itself is insufficient to account for the severity of autoimmune disease in this model, as demonstrated by analysis of yaa and lpr hybrids. Thus, B6.RIIB(-/-)/lpr mice are protected from disease progression, despite equivalent titers of ANA. In contrast, B6.RIIB(-/-)/yaa mice have significantly enhanced disease despite reduced ANA titers. Yaa modifies the specificity and thus the pathogenicity of the B6. RIIB(-/-) ANA, by converting them to antinucleolar antibodies. In addition to these known modifier pathways, we have discovered two novel, recessive loci contributed by the C57BL/6 genome that are required for the ANA phenotype, further indicating the epistatic properties of this SLE model.     

Evaluation of safety and efficacy of RNAi against HIV-1 in the human immune system (Rag-2(-/-)gammac(-/-)) mouse model

RNA interference (RNAi) gene therapy against HIV-1 by stable expression of antiviral short hairpin RNAs (shRNAs) can potently inhibit viral replication in T cells. Recently, a mouse model with a human immune system (HIS) was developed that can be productively infected with HIV-1. In this in vivo model, in which Rag-2(-/-)gamma(c)(-/-) mice are engrafted with human CD34(+)CD38(-) hematopoietic precursor cells, we evaluated an anti-HIV RNAi gene therapy. Human hematopoietic stem cells were transduced with a lentiviral vector expressing an shRNA against the HIV-1 nef gene (shNef) or the control vector. We observed normal development of the different cell subsets of the immune system. However, although initial transduction efficiencies were similar for both vectors, a reduced percentage of transduced human immune cells was observed for the shNef vector after establishment of the HIS in vivo. Further studies are required to fully evaluate the safety implications. When we infected the mature human CD4(+) T cells from the HIS mouse ex vivo with HIV-1, potent inhibition of viral replication was scored in shNef-expressing cells, confirming efficacy. When challenged with an shNef-resistant HIV-1 variant, equal replication was scored in control and shNef-expressing cells, confirming sequence-specificity of the RNAi therapy. We thus demonstrated that an antiviral RNAi-based gene therapy on blood stem cells leads to HIV-1-resistant T cells in vivo, an important proof of concept in the clinical development of RNAi against HIV-1.