老年人载脂蛋白E基因型与认知功能关系的研究

目的研究老年人中载脂蛋白E(Apolipoprotein E,ApoE)基因型的分布情况,分析Apo E基因型与认知功能的关系.方法用简易智能量表(Mini-Mental State Examination,MMSE)对316例60岁以上老龄人进行认知功能检查,根据MMSE得分将研究对象分为认知功能正常组和认知功能障碍组,分离其白血细胞基因组DNA,用聚合酶链反应(ploymerase chain reaction,PCR)技术扩增ApoE带有多态性位点的第四外显子片段,根据限制性酶切片段长度多态性分析(restriction fragment length polymorphism,RFLP)方法检测其基因型.结果两组人群中Apo Eε3/3基因型分布的百分比最大,其次为Apo Eε3/4基因型,而Apo Eε2/2基因型最小;ε2、ε3、ε4等位基因频率在认知功能障碍组和认知功能正常组分别为:0.095、0.560、0.345;0.146、0.699、0.155.认知功能障碍组ε4携带者百分比大于对照组(P<0.01),Apo Eε3/4基因型百分比也较认知功能正常组大(P<0.01).结论老年人群中,Apo Eε3/3为最常见的基因型,而ε2/2最少见;Apo Eε4为老年人认知功能障碍发病的危险因子.     

应用多重PCR法对广东地区HBV进行基因型(A-F)分型

目的建立乙型肝炎病毒(HBV)基因型(A-F)多重PCR分型方法,并利用该方法对广东地区HBV PCR阳性血清进行分型。方法将GenBank中114例HBV 全序列进行比较分析,找出每种基因型相对于其他五种基因型的独特序列,并根据这些独特序列设计出六对分别针对A-F基因型的特异引物。利用这六对引物建立HBV的多重PCR分型法。结果多重PCR与以前用PCR-限制温度长度多态型分析法的分型结果一致。对广州周边地区HBV携带者的初步分型结果显示,主要为B型和C型,分别占45.00%和38.75%,另外还有16.25%的D型。结论用多重PCR分型法准确易行,灵敏性高,便于推广应用。     

巢氏PCR—RFLP和多重PCR检测HBV基因型、基因亚型方法的比较

目的:建立HBV基因型和亚型的分型方法,并分析相关的两种方法特异性和敏感性。方法:采用型特异性引物的巢氏PCR-RFLP和六种主要的HBV型特异性引物和亚型特异性引物的多重PCR方法,分别检测了100例患者标本。结果:两法不一致率为50%(27/54)。型特异性引物巢氏PCR-RFLP法检测出B基因型41例(41%),C基因型25例(25%),B C基因型34例(34%),B j亚型3例(7.3%),Ba亚型为38例(92.7%),Cs亚型为21例(84%),Ce亚型为3例(12%),1例C型未分出亚型(4%)。多重PCR法检出B型18例(33.3%),C型7例(13%),B C混和型5例(9.3%),B2亚型2例(11%),C1亚型2例(28.5%)。巢氏PCR-RFLP法型检出率100%亚型检出率98.5%,多重PCR法型检出率55.6%,亚型检出率仅为16%,前者高于后者(P<0.05)。结论:巢氏PCR-RFLP鉴定HBV基因型、基因亚型较多重PCR敏感性高,重复性好,但耗时长,费用高。     

正常人群载脂蛋白E基因多态性的研究

为研究载脂蛋白E在正常汉族人体内的分布,应用聚合酶链-限制性片段长度多态性（PCR-RFLP）方法,共检测197例正常汉族人Apo E基因频率。结果纯合子ε_（3/3）基因型频率及ε_3等位基因频率最高,ε_2随年龄增长有增加趋势;证明该法检测快速、准确,分辨率高,并为Apo E生理功能的研究及临床应用打下基础。     

应用多重PCR法对广东地区HBV进行基因型（A—F）分型

目的：建立乙型肝炎病毒（HBV）基因型（A－F）多重PCR分型方法，并利用该方法对广东地区HBV PCR阳性血清进行分型。方法：将GenBank中114例HBV全序列进行比较分析，找出每种基因型相对于其他五种基因型的独特序列，并根据这些独特序列设计出六对分别针对A－F基因型的特异引物，利用这六对引物建立HBV的多重PCR分型法，结果：多重PCR与以前用PCR－限制温度长度多态型分析法的分型结果一致。对广州周边地区HBV携带者的初步分型结果显示，主要为B型和C型，分别占45．00％和38．75％，另外还有16．25％的D型，结论：用多重PCR分别法准确易行，灵敏性高，便于推广应用。     

Apo E基因检测在阿尔茨海默病早期诊断中的应用研究

目的研究Apo E基因多态性及其在阿尔茨海默病(AD)早期诊断中的应用。方法应用PCR-RFLP和聚丙烯酰胺凝胶电泳方法检测Apo E基因多态性。结果对照组与AD组之间,ε_2/ε_2、ε_3/ε_3、ε_4/ε_4、ε_2/ε_3和ε_2/ε_4基因型频率以及ε_2和ε_3基因频率间无显著性差异(P>0.05)。ε_3/ε_4基因型频率ε_4基因频率,AD组明显高于对照组(P<0.05)。结论 Apo E基因在连云港地区有遗传多态性,ε_4基因检测可用于AD的早期辅助诊断。     

荧光定量多重PCR检测13种高危型人乳头瘤病毒

目的:了解用荧光定量多重PCR检测13种高危型人乳头瘤病毒(HPV)的检测情况。方法:选取用基因芯片法检测的20份标本,再用荧光定量多重PCR法进行检测。结果:荧光定量多重PCR法包含的基因型检测结果与基因芯片法一致,没有包含的基因型检测结果显示阴性。结论:荧光定量多重PCR检测13种高危型HPV,方法简便准确,但也会漏检少量没有包含基因型的阳性结果。     

不同人群载脂蛋白E基因频率分布特征

目的 :研究江苏地区汉族人群载脂蛋白E(ApoE)基因型及等位基因频率 ,并与国内外不同人群比较分布特征。方法 :聚合酶链反应 -限制性片段长度多态性 (PCR -RFLP)方法检测 16 8例江苏地区无血缘关系汉族人群ApoE基因型。计算各基因型及等位基因频率。结果 :江苏地区汉族人群载脂蛋白E各基因型频率分别为 :ε2 / 2 =0 .6 0 % ;ε2 / 3 =11.90 % ;ε2 / 4 =1.2 0 % ;ε3 / 4 =10 .70 % ;ε3 / 3 =75 .0 0 % ;ε4 / 4 =0 .6 0 % ;各等位基因频率分别为 :ε2 =7.14% ;ε3 =86 .31% ;ε4 =6 .5 5 %。频率分布在不同年龄、性别无显著差异。与其他地区中国人群频率分布相似。中国人群ε3 等位基因频率明显高于欧美人群 ,而ε4等位基因频率明显低于欧美人群。结论 :江苏地区汉族人群载脂蛋白E等位基因频率分布与其他地区中国人群频率分布相似 ,但与欧美人群有显著差异。     

载脂蛋白E基因多态性与脑梗死的相关性

目的 探讨载脂蛋白E(apoE)基因多态性与脑梗死的相关性.方法 采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术,检测40例脑梗死患者以及50例健康人的apoE基因型,同时检测不同基因型患者总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)、载脂蛋白A(apoA)、载脂蛋白B(apoB)水平.结果 ε3/ε3为常见基因型.ε4等位基因频率脑梗死组 (20%)明显高于正常对照组 (6%)(P ＜0.01).apoEε4等位基因携带者总胆固醇、低密度脂蛋白、apoB水平比apoEε2、ε3携带者高 (P ＜0.05).结论 apoE基因多态性与脑梗死的发病有关,ε4基因可能为该病的危险因子,其发病机制可能是通过调控载脂蛋白E导致脂代谢紊乱.     

高脂血症患者载脂蛋白E基因测序分布及其与血脂谱改变的关系

目的 探讨高脂血症患者载脂蛋白E(ApoE)基因分布及其与血脂谱改变的关系.方法 采用全序列基因测序法检测212例高脂血症患者(高脂血症组)和100例健康体检者(对照组)的ApoE基因型,检测高脂血症患者血脂水平并分析其与ApoE基因多态性的相关性.结果 两组人群共检出6种基因型,分别是E2/2、E3/3、E4/4、E2/3、E2/4和E3/4;高脂血症患者E3/3基因型及ε3等位基因频率低于健康体检者,E3/4基因型及ε4等位基因频率高于健康体检者(P＜0.01);E3/4+E4/4基因型患者的总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)水平明显高于E3/3和E2/2+E2/3基因型者,高密度脂蛋白胆固醇(HDL-C)水平明显低于E2/2+E2/3基因型者,E3/3基因型患者的LDL-C水平明显高于E2/2+E2/3基因型者,差异均有统计学意义(P＜0.05).结论 ApoE基因型多态性与高脂血症存在一定的相关性,含ε4等位基因者具有更为明显的脂代谢紊乱特点.     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.