PAI-1启动子区4G/5G基因多态性与脑血管病相关性研究

目的初步探讨人类纤溶酶原激活物抑制物-1（plasminogenactivatorinhibitor,PAI-1）启动子区基因多态性与脑血管病的关系及其在脑血管病发病过程中的作用。方法通过多聚酶链反应（polymerasechainreaction,PCR）技术和发色底物法(ELISA),测定96例脑血管病病人,其中脑梗死（cerebralinfarction,CI）组65例,脑出血（cerebralhemorrhage,CH）组31例和60例对照组的白细胞PAI-1启动子区4G/5G多态性位点的基因型及血浆PAI-1活性。结果CI组血浆PAI-1活性明显高于其它二组,各组中均以纯合子4G/4G基因型患者的PAI-1血浆活性水平为最高,5G/5G基因型最低,杂合子4G/5G基因型居中;4G纯合子基因型与其它二型之间比较差异均有显著意义,4G/5G与5G/5G基因型之间比较差异无显著意义。CI组4G/4G纯合子型基因型与对照组（Controls）比较有显著性差异（P<0.05）,CI组基因型与CH组及CH组与对照组基因型比较均无统计学意义（P>0.05）。女性CI4G纯合子基因型患者血浆PAI-1活性与同型男性患者比较有显著性差异。结论纯合子4G/4G基因型可能是CI发病的危险因素之一,4G纯合子个体可能具有较高的CI发病倾向,尤其可能与女性CI发病相关。     

PAI-1启动子区插入/缺失(5G/4G)多态性与脑血管病的相关性研究

目的 初步探讨人类纤溶酶原激活物抑制物－1（plasminogen ativator inhibhitor-1,PAI-1)启动子区基因多态性与脑血管病的关系,及其在脑血管病发病过程中的作用。方法 通过多聚酶链反应技术和发色底物法（ELISA）,测定了96例脑卒中患者和60名健康对照者的白细胞PAI－1启动子区4G／5G多态位点的基因型及血浆PAI－1活性。结果 脑梗死（cerebral infarction,CI)组血浆PAI－1活性明显高于脑出血（cerabral hemorrhage,CH)组及对照组,脑梗死和脑出血组中均以纯合子4G／4G基因型患者的PAI－1血浆活性水平最高,5G／5G基因型最低,杂合子4G／5G基因型居中;4G纯合子基因型与其它二型之间比较差异无有显著性,4G／5G与5G／5G基因型之间比较差异无显著性。CI组4G／4G纯合子基因型与对照组比较差异有显著性（P＜0．05）,CI组基因型与CH组及CH组与对照组基因型比较差异均无显著性（P＞0．05）。CI组女性4G纯合子基因型患者血浆PAI－1活性与同型男性患者比较差异有显著性（P＜0．05）。结论 纯合子4G／4G基因型可能是CI发病的危险因素之一,4G纯合子个体可能具有较高的CI发病倾向,尤其可能与女性CI发病相关。     

RAI—1启动子区4G／5G基因多态性与脑血管病相关性研究

目的：初步探讨人类纤溶酶原激活物换制物－1（plasminogen activator inhibitor,PAI-1）启动子区基因多态性与脑血管病的关系及其在脑血管病发病过程中的作用。方法：通过多聚酶链反应（polymerase chain reaction,PCR）技术和发色底物法（ELISA），测定96例脑血管病病人，其中脑梗死(cerebral infarction,CI）组65例，脑出血（cerebral hemorrhage,CH）组31例和60例对照组的白细胞PAI－1启动子区4G／5G多态性位点的基因型及血浆PAI－1活性。结果；CI组血浆PAI－1活性明显高于其它二组，各组中均以纯合子4G／4G基因型患者的PAI－1血浆活性水平为最高，5G／5G基因型最低，杂合子4G／5G基因型居中；4G纯合子基因型与其它二型之间比较均有显著意义，4G/5G与5G／5G基因型之间比较差异无显著意义。CI组4G／4G纯合子型基因型与对照组（Controls）比较有显著性差异（P＜0.05),CI组基因型与CH组及CH组与对照组基因型比较均无统计学意义（P＞0.050。妇性CI 4G纯合子基因型患者血浆PAI－1活性与同型男性患者比较有显著性差异。结论：纯合子4G／4G基因型可能是CI发病的危险因素之一，4G纯合子个体可能具有较高的CI发病倾向，尤其可能与女性CI发病相关。     

儿童脓毒症纤溶酶原激活物抑制剂-1基因启动子区4G/5G多态性的研究

目的探讨纤溶酶原激活物抑制剂-1（PAI-1）基因启动子区单核苷酸插入或缺失（4G/5G）多态性与广州地区汉族脓毒症患儿的相关性,对脓毒症的发生、发展和临床预后的影响。方法选取2007年4-12月广州市妇女儿童医疗中心诊治的汉族脓毒症患儿为病例组,同期收集健康查体儿童为对照组。应用等位基因特异性扩增多聚酶链（AS-PCR）法对病例组和对照组行PAI-1基因启动子区4G/5G多态性检测和分析。采用基因计数法计算各组基因型频率和等位基因频率,χ^2检验分析比较两组人群各基因型的分布差异,计算OR值及其95%CI评估各基因型的风险。结果研究期间病例组纳入148例,对照组181名。病例组和对照组PAI-1基因启动子区4G/5G多态性的基因型和等位基因频率分布差异无统计学意义（χ^2=0.79,P〉0.05）。等位基因4G（χ^2=4.35,P〈0.05）及其纯合子（χ^2=4.44,P〈0.05）与脓毒症发展相关;携带等位基因4G患儿发展至重症脓毒症的风险比5G高,OR=4.05（95%CI：1.09-15.08）,4G/4G纯合子患儿发展至重症脓毒症的风险比其他基因型高,OR=4.57（95%CI：1.11-18.78）。等位基因4G（χ^2=9.17,P〈0.05）及其纯合子（χ^2=7.35,P〈0.05）与脓毒症病死率相关,携带等位基因4G患儿脓毒症病死风险较5G高,OR=4.30（95%CI：1.50-12.29）,4G/4G纯合子患儿脓毒症病死风险较其他基因型高,OR=3.14（95%CI：1.49-6.61）。结论PAI-1基因启动子区4G/5G多态性与广州地区汉族脓毒症患儿进展及预后相关,等位基因4G及其纯合子是其高危遗传因素;PAI-1基因启动子区4G/5G点多态性与脓毒症的易感性无关。     

PAI-1基因4G4G基因型与IgA肾病易感性及临床表现的关系

目的探讨PAI-1基因启动子区4G／5G多态性与IgA肾病的发生、进展和临床表现的关系。方法收集IgA肾病患者的临床资料；应用PCR-限制性片段长度多态技术分析296例IgA肾病患者和310名健康人的PAI-1基因4G／5G多态性；分析PAI-1基因4G／5G多态性与IgA肾病的发生与临床表现及病理改变的关系。结果（1）PAI-1基因4G4G，4G5G，5G5G基因型频率在IgA肾病组和正常对照组分别为0．33、0．19、0．48和0．3、0．23、0．47，两组之间差异无统计学意义（X^2=1．63，P〉0．05）；（2）4G4G纯合子基因型频率在病理Lee氏分级Ⅲ级以下组（A组）和Ⅳ～Ⅴ级组（B组）分别为0．39和0．28，（X^20=7．86，P〈0．05）。（3）按基因型分组，4G4G组IgA肾病患者的肌酐清除率明显低于非4G4G组；4G4G组患者的血清甘油三酯水平明显高于5G5G组；4G4G组患者高甘油三酯血症发生率明显高于4G5G组（P〈0．05）。结论PAI-1基因启动子区4G／5G多态性不是IgA肾病发生的易感因素，但可能是IgA肾病病情加重的危险因子。     

复发性流产与β-纤维蛋白原基因-455G／A多态性关系的研究

目的探讨血浆纤维蛋白原（Fg）及βFg-455G／A基因多态性与山西地区汉族妇女复发性流产的关系。方法应用聚合酶链反应-限制性片段长度多态性（PCR—RFLP）方法，检测复发性流产组30例和对照组30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X2检验，各基因型构成比和等位基因频率在两组间均无显著性差异（P〉0.05）；复发性流产纽血浆Fg水平（2.31±0．57g/L）较对照组（3.08±0．57g/L）显著降低（P〈0．05）。结论本研究发现血浆纤维蛋白原水平降低是复发性流产的危险因素，βFg-455G／A基因多态性不会影响复发性流产血浆纤维蛋白原水平。     

PAI-1基因4G/5G多态性与特发性肺纤维化的相关性研究

目的探讨纤溶酶原激活剂抑制物-1（plasminogen activator inhibitor-1,PAI-1）基因启动子区4G/5G多态性与特发性肺纤维化（IPF）的相关性。方法应用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）分析,检测42例IPF患者和164例正常对照组人群PAI-1基因4G/5G多态性。结果PAI-1基因4G/4G、4G/5G、5G/5G基因型频率分布,IPF组分别为31.0%、50.0%、19.0%,对照组分别为17.1%、54.9%、28.0%;4G和5G等位基因频率,IPF组分别为0.560和0.440,对照组分别为0.445和0.555;4G/4G基因型频率IPF组显著高于对照组（P〈0.05）。与4G/5G和5G/5G基因型比较,携带4G/4G型个体发生IPF的风险增加2.18倍,95%CI：1.02～4.68（P〈0.05）。结论PAI-1基因4G/5G多态性与IPF的发病相关,纯合子4G/4G基因型可能是IPF发病的重要危险因素之一。     

PAI-1启动子区4G/5G基因多态性、ACE插入/缺失多态性与脑卒中的相关性

目的探讨纤溶酶原激活物抑制物-1(PAI-1)启动子区基因多态性和血管紧张素转换酶(ACE)插入/缺失多态性与脑卒中的关系。方法 PCR检测203例脑卒中患者和139名健康对照者PAI-1基因启动子区4G/5G多态性、ACE基因插入/缺失多态性,同时应用比色法测定血清ACE活性,发色底物法测定PAI-1活性。结果脑梗死(CI)组PAI-1活性(0.769±0.163 AU/mL)、ACE活性(43.42±14.36 U/L)明显高于对照组(0.652±0.116 AU/mL和31.28±8.64 U/L,P<0.01);CI组PAI-I基因4G纯合子、ACE D/I基因DD纯合子比例明显高于对照组(P<0.01);PAI-I基因4G/4G基因型与ACE基因D/D基因型对CI发病可相互协同作用(P<0.01)。结论 PAI-1基因4G/4G基因型和ACE基因D/D基因型均可能是CI发病的危险因素,且具有协同作用。     

超声影像学与分子生物学在复发性自然流产的联合应用

目的探讨纤溶酶原激活剂抑制物-1(plasminogen activator inhibitor-1,PAI-1)基因4G/5G多态性和早期妊娠子宫动脉舒张期切迹与复发性自然流产的关系,为早期诊断和治疗复发性自然流产提供依据。方法应用聚合酶链反应-限制性片段多态性(PCR-RFLP)分析,对61例反复自然流产患者(病例组)和52例正常妊娠对照组(对照组)做了PAI-1基因4G/5G多态性分析,并应用彩色多普勒超声检测子宫动脉舒张期切迹(uterine artery diastolic notches depth,ND)结果1.复发性自然流产组有ND切迹者占54.1%(33/61),正常妊娠对照组为34.62%(18/52),二者差异有显著性意义(χ2=4.30,P<0.05,OR值为2.23,95%CI 1.04-4.77。2.携带4G/4G基因型患者其子宫动脉血流循环阻力有增加趋势,(χ2=16.08,P<0.01,OR值为6.0,95%CI 2.38-15.12。结论1.子宫动脉血流循环阻力增加与复发性自然流产的发病有关联。2.PAI-1基因4G/4G型个体,同时有ND者自然流产的风险为对照组的4.25倍...     

RANTES基因-28C／G单核苷酸多态性与子宫内膜异位症遗传易感性的研究

目的探讨调节正常T细胞表达和分泌活性因子（regulated on activation, normal T cell expressed and secreted,RANTES）基因启动子区-28C／G单核苷酸的多态性与广东籍汉族患者子宫内膜异位症的关系。方法应用聚合酶链反应-限制性酶切片段长度多态性技术（PCR—RFLP）并进行基因测序的方法检测广东籍汉族子宫内膜异位症患者59例（内异症组）,非子宫内膜异位症患者49例（对照组）,比较分析各组间基因型频率和等位基因频率。结果RANTES基因启动子区-28C／C基因型在子宫内膜异症组及对照组分布频率分别为81．36％、81．63％,C／G基因型分布频率分别为18．64％、18．37％;两组间的基因型分布频率比较差异无显著性（P〉0．05）。RANTES基因启动子区-28位点C等位基因型在内异症组及对照组中的分布频率分别为90．68％、90．82％,G等位基因型分布频率分别为9．32％、9．18％,两组间等位基因型频率比较差异无统计学意义（P〉0．05）。结论在广东籍汉族妇女中,RANTES基因启动子区-28C／G单核苷酸多态性与子宫内膜异位症遗传易感性可能无关联。     

Hutchinson-Gilford progeria syndrome with G608G LMNA mutation

Hutchinson-Gilford progeria syndrome (HGPS) is a rare condition originally described by Hutchinson in 1886. Death result from cardiac complications in the majority of cases and usually occurs at average age of thirteen years. A 4-yr old boy had typical clinical findings such as short stature, craniofacial disproportion, alopecia, prominent scalp veins and sclerodermatous skin. This abnormal appearance began at age of 1 yr. On serological and hormonal evaluation, all values are within normal range. He was neurologically intact with motor and mental development. An echocardiogram showed calcification of aortic and mitral valves. Hypertrophy of internal layer at internal carotid artery suggesting atherosclerosis was found by carotid doppler sonography. He is on low dose aspirin to prevent thromboembolic episodes and on regular follow up. Gene study showed typical G608G (GGC- > GGT) point mutation at exon 11 in LMNA gene. This is a rare case of Hutchinson-Gilford progeria syndrome confirmed by genetic analysis in Korea.     

Direct analysis of the genes encoding G proteins G alpha T2, G alpha o, G alpha Z in ADHD.

We have followed up the extensive replicated evidence that the dopamine DRD4 receptor is involved in the aetiology of ADHD by undertaking direct analysis of genes encoding other proteins in this effector system. We prioritised the genes encoding G protein alpha subunits G alpha(T2), G alpha(o), G alpha(Z) as these have been shown to transduce the effects of ligand binding at DRD4. We screened the exons of all three genes for sequence variation in 28 unrelated subjects with ADHD and identified 13 novel polymorphisms. All were tested for possible association with ADHD using a combination of pooled and individual genotyping. The results of our study do not suggest that polymorphisms in these genes contribute to susceptibility to ADHD.     

Fission yeast enters the stationary phase G0 state from either mitotic G1 or G2

Summary The cell cycle of Schizosaccharomyces pombe in continuous culture is controlled at two steps, one which limits the transition from G₁ to S phase and the other which determines the timing of cell division. We have investigated, by means of flow-cytofluorometry, the cell cycle characteristics of nutritionally starved cells in stationary phase. Cells were shown to become arrested in either G₁ or G₂, in ratios which depended on the composition of the growth medium. G₁ and G₂ stationary phase cells share certain properties. (1) They become relatively resistant to heat shock. (2) They can reenter the cell cycle after subculture into fresh medium. (3) The G₁ and G₂ arrested populations have equal long-term viability in stationary phase. (4) Both populations require the activity of the cdc2 ⁺ gene for reentry into the cell cycle. We suggest that cell cycle arrest in stationary phase is regulated by the activity of the same G₁ and G₂ controls which limit the rate of cell cycle progression in continuous culture. The data demonstrate that in fission yeast the transition from G₁ to S phase does not mark a point of commitment to the completion of the cell cycle.     

G. Holzknecht

Ohne Zusammenfassung     

Maturation of Hantaan virus glycoproteins G1 and G2.

Hantaan virus-infected Vero E6 cell lysates were used for immunoprecipitation with monoclonal antibodies against glycoprotein G1 (MAbG1) or G2 (MAbG2). When cell lysates were prepared with buffer containing nonionic detergent, both G1 and G2 glycoproteins were precipitated with either MAbG1 or MAbG2. In contrast, when cell lysates were prepared with a buffer containing ionic detergents MAbG1 precipitated only glycoprotein G1 and MAbG2 precipitated only glycoprotein G2. Heterodimers and possibly higher oligomeric forms of the glycoproteins were detected on nonreducing SDS-polyacrylamide gels only after chemical cross-linking and immunoprecipitation with either MAbG1 or MAbG2. In order to determine the sites of Hantaan virus glycoproteins maturation and the G1-G2 complex formation, infected cells were treated with inhibitors that prevent specific steps of oligosaccharide processing. Furthermore, glycoproteins G1 and G2 immunoprecipitated from infected cell lysates or from isolated virus particles were tested for sensitivity to endoglycosidase H, endoglycosidase F, and endoglycosidase D. The results of these experiments show that maturation of both G1 and G2 takes place in the endoplasmic reticulum (ER). Furthermore, G1-G2 complex formation occurs in the ER as well, since the two glycoproteins co-precipitated with either MAbG1 or MAbG2 from infected cell lysates treated with brefeldin A and prepared with buffer containing nonionic detergent.     

The G syndrome-additional observations

We are reporting the second lethal case of G syndrome occurring in a female. The developmental defects in this patient included posteriorly angulated auricles, bifid tip of tongue with a long frenulum, hypoplasia of the epiglottis and larynx, rocker bottom feet, and hypertrophied labia majora and clitoris. Additional anomalies not previously reported in the G syndrome were circumvallate placenta, and incompletely perforated hymen.