生物素-链霉抗生物素蛋白硷性磷酸酶体系标记检测DNA探针

本室采用近年发展的生物素—链霉抗生物素蛋白体系检测特异互补的核酸序列,先经缺口翻译制备生物素DNA探针,分子杂交后,再与链霉抗生物素蛋白—生物素—硷性磷酸酶复合物反应和显色,检测限度约50pg靶DNA,并将其结果与~(35)S标记相同的探针进行了比较。     

生物素探针分子杂交检测92例血清乙肝病毒DNA的结果分析

血清HBV—DNA的检测是目前诊断乙型肝炎,判断HBV复制和患者肝脏病变和予后的最重要指标之一。1992年6月,我们引进北京医科大学肝病研究所研制推出的生物素探针分子杂交实验(Bio—DNA),对92例不同人群乙肝病毒核酸(HBV—DNA)进行了检测,现将结果报告如下:     

新型TaqMan-MB荧光探针测定古DNA模板含量

目的:设计荧光检测探针,对古DNA样本中的线粒体DNA片段进行定量分析,并对其检测效果进行评价。方法:利用分子信标的茎环结构和TaqMan的降解作用设计一种新型的TaqMan-Molecular Beacon荧光检测探针,与实时PCR仪联用,对古DNA模板进行定量检测。结果: 5个古代样本中3个测得拷贝数,1个无法观察到原始模板量,1个样本无原始模板。结论: TaqMan-MB荧光探针集中了分子信标和TaqMan的技术优势,对古DNA模板具有很好的定量检测效果。     

HBV—DNA分子杂交技术的建立及其临床方面的应用

本文采用目前最灵敏的HBV—DNA分子杂交技术,利用α—~(32)P和生物素标记DNA做为探针进行乙型肝炎的研究。对230例HBsAg阳性的携带者进行检测HBV—DNA阳性者占26.5%。230例中HBeAg占63%,抗—HBe占25.2%。而HBV—DNA阳性的血清中HBeAg阳性占86.9%为阳性。抗—HBe中只有8.1%。两者有显著差异。本文还对HBV—DNA与HBsAg免疫复合物的关系以及乙肝疫苗的检测有一些新的评价。     

功能性硫化锌纳米荧光探针与DNA的相互作用

目的:研究L-半胱氨酸包裹的ZnS纳米荧光探针与核酸的相互作用及其机理,建立DNA的定量分析方法.方法:通过考察溶液pH值、缓冲液用量及粒子浓度等影响因素,在最佳试验条件下,利用在pH值近中性时DNA对纳米荧光探针具有猝灭作用的特性,采用荧光光谱法进行研究.结果:当DNA浓度为20～600 μg/L时,荧光强度与DNA的浓度呈良好的线性关系,检测限为0.032 μg/L.结论:该方法简单,快速,灵敏度高.DNA与L-半胱氨酸-ZnS纳米粒子存在静电作用.     

小鼠PTA1(CD226)胞膜外两个结构域同时与小鼠Tage4(CD155)的相互作用

目的:确定mPTA1分子与mTage4参与相互作用的结构域,以深入研究这两种分子的功能. 方法: 扩增编码mTage4全长分子和mPTA1,hICAM-1分子不同结构域的DNA序列,构建含mTage4,mPTA1不同结构域编码序列的截短体、mPTA1/hICAM-1荧光蛋白嵌合体分子基因的真核表达载体,并转染COS-7真核细胞,通过激光共聚焦扫描显微镜研究这两种分子间的相互作用. 结果: DNA序列测定证实成功构建了含mTage4全长分子和mPTA1截短体、mPTA1/hICAM-1嵌合体分子基因的真核表达载体,激光共聚焦扫描显微镜观察结果表明mPTA1截短体分子和mPTA1/hICAM-1嵌合体分子不能与mTage4有效结合,而mPTA1全长分子能与mTage4相互作用. 结论: mPTA1分子胞膜外区两个结构域同时参与了与mTage4的相互作用.     

HBV—DNA斑点杂交技术在肝病血清学诊断中的应用价值

<正> 近年来,由于HBV—DNA已能用基因工程的方法克隆化并制备成探针,HBV—DNA斑点分子杂交技术在乙肝研究中广为应用。 HBV—DNA则为检测乙肝病毒直接的指标,较其他方法反映的结果更确切更可靠,为乙型肝炎诊断提供一项最敏感的检测手段。我院1985～1988年检测500例各型肝炎患者血清,现将结果报告如下。     

金电极上自组装DNA传感器的研究

目的:在金电极表面自组装DNA,制备DNA传感器.方法:以双层组装技术,先进行-SH化合物自组装,得到自组装单分子层,再在SAM上吸附固定DNA,制备DNA修饰电极,用电化学分析法对其组装过程及活性进行表征.结果:基于构建的新型电极,表明双层膜组装方法是组装DNA分子的有效手段,可进一步研究DNA的电化学特性及与其他分子的相互作用.结论:该电极显示良好灵敏度和稳定性.     

基因探针技术与平板免疫溶血试验检测ETEC—LT的比较

本文报告从腹泻患者检出237株大肠艾希氏菌,用PIHT法与LT—DNA探针技术进行ETEC—LT检测比较。结果LT—DNA探针技术检出率(13.5％)明显高于PIHT法(4.7%),未发现PIHT法检出阳性而LT—DNA探针阴性者。认为LT—DNA探针特异性强,敏感度高,标本又可直接点膜检测,不需作系统培养鉴定,适用流行病学调查大批量的标本检测等优点。     

银川地区HBsAg阳性人群中HBV—DNA的分布

以非放射性(Dig—HBV—DNA 探针)分子杂交技术,检测了银川地区405例 HBsag携带人群中的 HBV—DNA。阳性率为34.81%(男性40.32%高于女性30.14%,P<0.05),且随年龄的增长而极急速地下降(P<0.005),回汉民族间无明显差异。HBsAg 的滴度与HBV—DNA 的检出率呈正相关(γ_s=0.999,P<0.01)HBsAg 的滴度至≥1:256时,HBV—DNA 的检出率可高达100%。作者等认为;以 HBsAg 的定量滴度来判定其传染性有一定的参考价值;滴度在1:64～1:128时需加强治疗管理,≥1:256时应视为病毒复制的标志,并具有较强的传染性。     

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.