生物素—链霉亲和素系统在检测HBeAg中的比较

应用链霉亲和素和生物素系统发展成的酶连接免疫吸附测定（ELISA）改良法检测HBeAg，取得较满意效果。标记链霉亲和素-辣根过氧化物酶（SA-HRP）时，两的重量比以（0.5-0.8):1为佳，浓度以7μg/ml为最理想；SA-HRP的反应时间以30min为适宜。本改良法与生物素亲和素ELISA（BA）法、普通ELISA法检测HBeAg的灵敏度比较。其相对灵敏度高于后二；用Abbott试剂作为标准。检测HBeAg100人份，其阳性率高于BA法和普通ELISA法，提示链霉亲和素FLISA（BSA）法可提高方法的特异性及灵敏度。     

生物素－链霉亲和素系统在检测HBeAg中的比较

应用链霉亲和素和生物素系统发展成的酶连接免疫吸附测定（ＥＬＩＳＡ）改良法检测ＨＢｅＡｇ，取得较满意效果。标记链霉亲和素－辣根过氧化物酶（ＳＡ－ＨＲＰ）时，两者的重量比以（０．５～０．８）：１为佳，浓度以７μｇ／ｍｌ为最理想；ＳＡ－ＨＲＰ的反应时间以３０ｍｉｎ为适宜。本改良法与生物素亲和素ＥＬＩＳＡ（ＢＡ）法、普通ＥＬＩＳＡ法检测ＨＢｅＡｇ的灵敏度比较，其相对灵敏度高于后二者；用Ａｂｂｏｔｔ试剂作为标准，检测ＨＢｅＡｇ１００人份，其阳性率高于ＢＡ法和普通ＥＬＩＳＡ法。提示链霉亲和素ＥＩ，ＩＳＡ（ＢＳＡ）法可提高方法的特异性及灵敏度。     

链酶亲和素-生物素ELISA检测人心肌肌钙蛋白T

目的 建立人心肌肌钙蛋白T (cTnT)链酶亲和素-生物素（SAB）双抗体夹心酶联免疫分析（ELISA）方法,诊断急性心肌损伤性疾病。方法 以固相化单抗3F7捕捉样品中cTnT,生物素化3H12单抗为检测抗体,加入SA-HRP,显色。根据已知不同浓度标准cTnT显色后D(l)值的标准曲线,检测待测样品cTnT含量。结果 cTnT的最低检测值为0.15 ng/ml,批内变异系数为3.8%,批间变异系数为11.2%,回收率为98%。35例急性心肌损伤患者cTnT含量为(0.68±0.17) ng/ml,明显高于52例正常人对照组(0.20±0.03) ng/ml(P<0.01)。结论 本方法灵敏、特异、稳定,可用于临床急性心肌损伤疾病的诊断。     

固相生物素－亲和素ELISA检测HIV抗体方法的建立与应用

采用生物素化ＨＩＶ合成肽抗原与预吸附在ＥＩＡ反应板上的亲和素结合，建立检测ＨＩＶ抗体的固相生物素－亲和素ＥＬＩＳＡ方法。经ＨＩＶ（１＋２）抗体参考血清Ｐａｎｅｌ检定，其灵敏度（１３／１３）１００％，特异性（２８／２８）１００％，批内变异系数＜７％，批间变异系数＜１０％。１０５６８例确认血清标本验证，阳性检出率１００％，阴性总符合率９９．８％，假阳性率０．２％。与常规间接ＥＬＩＳＡ法比较，本法操作简便，试验所需时间短，灵敏度、特异性和稳定性提高明显     

PCR—EIA法检测端粒酶活性及其临床应用

目的：探讨用PCR－EIA法检测人端粒酶活性及其临床应用。方法：利用kim法处理细胞和组织标本及扩增端粒酶产物，引物TS用生物素标记，CX用地高辛标记，PCR产物与包被有链亲和素的酶标板结合，并与酶标抗地高辛抗体反应，用TMB显色。结果：PCR－EIA法与TRAP－银染色法有很好的相关性，批内变异系数CV为4．138％，在30例各种恶性肿瘤组织中端粒酶活性检出率为90％，28例癌旁组织中为7．1％。结论：该法具有较好的灵敏度与重复性，无同位素污染，较银染色法更为简便、快速，检测成本低，无需特殊仪器，适合于各级医院推广。     

链酶亲和素—生物素ELISA检测人心肌肌钙蛋白T

目的 建立人心肌肌钙蛋白Ｔ（ｃＴｎＴ）链酶亲和素－生物素（ＳＡＢ）双抗体夹心酶联免疫分析（ＥＬＩＳＡ）方法。诊断急性心肌损伤性疾病。方法 以固相化单抗３Ｆ７捕捉样品中ｃＴｎＴ,生物素化３１１１２单抗为检测抗体,加入ＳＡ－ＨＲＰ,显色。根据已知不同浓度标准ｃＴｎＴ显色后Ｄ（λ）值的标准曲线,检测待测样品ｃＴｎＴ含量。结果 ｃＴｎＴ的最低检测值为０．１５ｎｇ／ｍｌ,批内变异系数为３．８％,批间变异系数     

生物素-链霉亲和素-过氧化物酶法快速诊断呼吸道合胞病毒感染

目的：建立检测下呼吸道感染婴幼儿粘膜细胞呼吸道合胞病毒（RSV）抗原的方法。方法：以生物素标记兔抗RSV抗体，鼻咽部粘膜细胞涂片与该抗体作用后加链霉亲和素-过氧化物酶反应，二氨基联苯胺显色，显微镜下观察结果。结果：RSV抗原总阳性率为41．6％，临床典型和可疑病例抗原检出率分别为47．9％和44．9％，临床不典型者阳性率12．5％（X2=7．9，P＜0．05）。阳性标本中细胞着成棕色，易与阴性标本相区别。结论：本法简便、快速，具有一定敏感性和特异性，适合于基层推广应用。     

生物素-链霉亲和素技术一步法检测淋球菌抗原的研究

目的 :建立一种灵敏、快速诊断淋球菌感染的酶免疫测定法。方法 :检测淋球菌抗原的生物素 -链霉亲和素酶联免疫一步法。结果 :36例临床淋球菌标本中淋球菌抗原含量 1 0 8.47± 5 1 .39μg/ L ,阳性率 97.2 % ,显著高于非淋球菌标本的 4.71± 2 .1 3μg/ L ,阳性率 0 % (P均 <0 .0 1 ) ,本法检测灵敏度 0 .6 2 5 μg/ L,特异性 1 0 0 % ,敏感性 97.2 %。结论 :该法可作为灵敏、快速诊断淋球菌的新的辅助方法。     

半巢式聚合酶链反应-微孔板杂交法检测沙眼衣原体

目的：应用半巢式聚合酶链反应－微孔板杂交法（半巢式PCR－MPH）检测泌尿生殖道沙眼衣原体（Ct)。方法：分别用质粒和主要外膜蛋白基因引物进行扩增，将下游引物用生物素标记，经半巢式PCR扩增Ct主要外膜蛋白基因后，扩增产物被包被有链霉亲和素的微孔板捕获，经碱变性后与标有荧光素的特异性探针进行杂交，然后通过辣根酶标记的抗荧光素抗体与杂交分子反应，经过酶底物显色，通过测定吸光度来判断结果。结果：主要外膜蛋白基因引物半巢式PCR较质粒引物PCR更敏感，比半巢式PCR产物酶切后电泳法高10倍，半巢式PCR－MPH法特异性强，该法重复法好，批内CV值＜10％，批间CV值＜15％，该法在包被浓度为4mg/L，产物1：20稀释，与微孔板结合20min后加入10pmol/mL探针杂交30min,用1：3000稀释的酶显色条件下有最佳结果，结论：该法操作简便，敏感性和特异性均高，无放射性和EB染料污染，适于临床样本的常规检测。     

人端粒酶亲和纯化方法的建立及其初步鉴定

建立一种亲和纯化人端粒酶的手段,方法酶切鉴定人端粒酶ＲＮＡ质粒,所切下的ｈＴＲｃＤＮＡ片段用生物素标记后与链霉亲和素琼脂糖珠耦连,用高端粒酶活性的胃肿瘤细胞提取液与之反应,利用端粒酶核糖核蛋白的特性,形成链霉亲和素琼脂糖珠－生物素标记ｈＴＲｃＤＮＡ－ｈＴＲ－端粒酶蛋白组份复合物,核酸解离液洗脱后获得亲和纯人端粒酶。     

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.     

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.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.