河南省2003年医药卫生科研进展

1．河南汉族群体中短串联重复序列的遗传多态性研究及应用：人类基因组是一个结构十分稳定的体系，同时又是一个变异的体系。在长期的进化过程中，基因组DNA的序列不断地发生变异，导致了不同种族、群体和个体间基因组的差异和多态性。随着分子生物学技术的不断发展，检测基因组DNA多态性已成为可能。河南地处中原，人口众多，河南汉族群体     

河南省2003年医药卫生科研进展

一、基础医学研究 1.河南汉族群体中短串联重复序列的遗传多态性研究及应用:人类基因组是一个结构十分稳定的体系,同时又是一个变异的体系。在长期的进化过程中,基因组DNA的序列不断地发生变异,导致了不同种族、群体和个体间基因组的差异和多态性。随着分子生物学技术的不断发展,检测基因组DNA多态性已成为可能。河南地处中原,人口众多,河南汉族群体的遗传学特征对中国北方汉族人群有较好的代表性。河南省     

克隆波尔山羊的微卫星DNA鉴定

目的:利用微卫星DNA技术进行体细胞克隆后获得个体的鉴定.方法:提取正常山羊、供核波尔山羊、本地受体山羊及克隆个体的基因组DNA,通过设计微卫星DNA多态PCR引物扩增微卫星DNA序列并对之进行微卫星DNA鉴定.结论:通过微卫星DNA序列的扩增,证明克隆个体为供核波尔山羊的克隆.     

人类基因组微卫星DNA多态及其法医学应用

人类基因组是指人类细胞的DNA 分子中所包含的储藏有人体全部遗传信息的一整套基因它包括细胞核所构成的基因组和线粒体DNA 所构成的基因组两部分细胞核基因组规模庞大结构复杂包含了人类基因组的绝大部分基因其DNA 分子总长度可达3 10 6 Kb 基因总数约5 ~10 万个而线粒体基因组是一个结构简单的小基因组DNA 分子总长度约1 6 .6 Kb 基因数仅37 个人类基因组是一个十分稳定的体系不同民族不同群体的个体有相同数目的基因也有基本相同的核苷酸序列正是基因组结构的这种稳定性保证了人类作为一个物种的稳定性和同一物种向的共同性然而人…     

钩端螺旋体基因组大小的初步研究

作者采用脉冲场凝胶电泳技术,对不同属、群(型)的5株钩端螺旋体基因组DNA分子量进行了研究,并用限制性内切酶Not I对基因组DNA进行分析。结果表明:钩端螺旋体基因组大小为2000kb;各株钩体基因组大小无明显差异;限制性内切酶Not I可将钩体基因组消化为11个大片段;钩体基因组DNA被Not I消化后电泳图谱与细菌基因组DNA图谱有很大差异。     

实时荧光定量PCR检测精子线粒体DNA的提取效率

目的：测定宝生物工程有限（大连）公司和上海杰美基因医药科技有限公司两种试剂盒精子线粒体DNA提取的效率,以及核基因组DNA的残留。方法：CASA法计数精子数量,试剂盒提取线粒体DNA,TaqMan法荧光实时定量PCR,利用标准曲线测定线粒体基因Cox-I片段、核基因β-actin片段的拷贝数,计算出精子线粒体DNA的提取效率和核基因组DNA的残留。结果：两种试剂盒精子线粒体DNA提取效率分别是：0.44%±0.03%和6.19%±0.17%。核基因组DNA的残留率分别为4.71%±0.76%和38.1%±1.97%。结论：后者线粒体DNA提取效率高一个数量级,但核基因组DNA的残留率也高一个数量级。     

山医群体中国地鼠近交E家系RAPD分析

目的　分析山医群体中国地鼠E家系的遗传纯度。方法　应用经过筛选的 3 1条随机引物对中国地鼠E家系 12只个体基因组DNA进行RAPD扩增 ,计算近交个体间的相似系数。结果　所有样品的相似系数0 943 1到 0 9978之间变动 ,平均相似系数为 0 9749,聚类分析得到了这些个体的同源树资料。结论　山医群体E家系有较高的遗传纯度     

线粒体DNA突变及其与衰老的关系

真核生物基因组可以分为核 DNA ( n DNA,nuclear DNA)和线粒体 DNA ( mt DNA,mitochondrial DNA)二部分。哺乳类动物的mt DNA大小相近 ,基因的组织和定位也相似。人mt DNA由 1 65 69bp组成 ,是一个双链闭环分子 ,存在于线粒体基质中。它含有 37个基因 :2个r RNA( 1 2 s、1 6sr     

DNA指纹图技术对国内SD大鼠遗传距离的分析

目的 通过DNA指纹技术分析国内封闭群SD大鼠遗传背景分布情况。方法 采用JL-02多位点探针的DNA指纹技术，对国内最具有典型的6个地区的9个SD大鼠群体内和群体间进行分析比较。结果 同一群体不同个体间SD大鼠遗传距离主要在0．3-0．5之间；不同群体间SD大鼠DNA指纹图带的遗传距离主要分布在0．5—0．6之间。结论 DNA指纹图较好地反映了封闭群SD大鼠群体内和群体间动物个体的遗传本质及遗传背景，具有良好的多态性。     

河北汉族人群D2S1328基因座的遗传多态性研究

基因组DNA多态性的研究,对人类进化和群体遗传提供了更为科学和可靠的依据。短串联重复序列（short tandem repeat,STR)属微卫星DNA序列,重复单位2－7bp。STR位点在人类基因组中含量丰富,它是绘制基因组物理图谱和遗传连锁分析的理想标记。近年来,随着STR－PCR技术的进一步发展,解决了微量检材的分析问题,扩展了DNA分析在遗传学、人类学以及在法医学领域的应用。作者从正常健康人血液中提取DNA,采用聚合酶链反应（polymerase chain reaction,PCR）技术和聚丙烯酰胺凝胶电泳技术,并结合银染的方法,对中国河北省181名汉族无关个体进行了调查,首次获得了D2S1328基因座群体遗传数据,结果报告如下。     

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.