新型漏声表面波生物传感器液相检测条件的实验研究

目的探讨漏声表面波生物传感器液相检测系统的影响因素.方法构建检测和参比双路延迟线,并观察在不同影响因素如离子浓度、缓冲液pH值及不同粘度的甘油PBS溶液中两路延迟线的相位变化规律.结果检测延迟线声表面波信号的相位值随溶液粘度、离子浓度、pH值的增大而发生明显的变化,而参比通道的相位曲线变化不明显.＃结论成功构建了漏声表面波生物传感器液相检测检测技术平台,并探索了溶液的粘度、离子浓度、pH等因素对传感器的影响.     

压电石英晶体传感器金膜表面亲和素固定研究

目的 研究压电石英晶体传感器金膜表面亲和素固定的反应趋势及最佳条件。方法 用3,3＇-二巯基丙酸自组装方法将亲和素固定在压电石英晶体传感器金膜表面,比较不同浓度、不同pH值条件下亲和素固定引起的频率变化。结果 经单因素方差分析,磷酸盐缓冲液pH值为6．2时亲和素固定引起的频率变化与pH5．8、6．6时差异无统计学意义（P〉0．05）,但pH值为6．2时频率下降有最大均值．应用pH7．0、7．4溶液进行亲和素固定时频率改变显著减小（P〈0．01）。浓度为0．2mg／ml时亲和素固定已达饱和。结论 选用pH6．2的0．2mg／ml时亲和素进行压电石英晶体传感器金膜修饰。     

卵磷脂双层膜上胆红素的吸附行为

目的:研究胆红素的吸附行为,探索最佳分析条件。方法:利用石英晶体微天平研究磷酸盐缓冲溶液(PBS)中胆红素在卵磷脂双层膜的吸附行为,石英晶体震荡频率下降越多,表示吸附量越多。考察了溶液pH值、离子强度和温度的影响,并与巯基苯胺膜和巯基苯胺/卵磷脂修饰膜进行了比较。结果:溶液pH为8.0时,频率变化-780Hz;溶液中离子强度增加一倍,频率变化-76Hz;10℃、28℃时,频率变化分别为-709Hz和-374Hz。结论:卵磷脂双层膜在碱性条件下有最大吸附;离子强度也会增加胆红素在膜上的吸附,但是影响没有其它因素大;温度会显著的影响胆红素的吸附,低温时吸附量明显增加;不同膜上胆红素的吸附量不同,其中硫醇/卵磷脂修饰膜上最大。     

pH值对尿酸溶解度影响的实验研究

目的:探讨室温下(20～22℃)pH值对尿酸(uric Acid, UA)溶解度的影响,为深入了解体内UA的溶解与析出提供理论依据。方法:分别将UA溶于NaOH溶液和PBS溶液中制备母液,然后将母液配置成不同浓度和pH值的UA溶液;相同pH值下的UA溶液,取最低浓度1 000μmol/L为标准UA溶液,经紫外分光光度计检测其吸光度值,每个样本检测3次,取均值建立标准曲线方程。分别于0 h和24 h,检测其它不同浓度和pH值的样本溶液,重复3次。结果:(1)在NaOH溶液中,分别在0 h和24 h时对1 000、2 000、3 000和4 000μmol/L样本溶液中UA的溶解度进行检测,结果显示随pH值的升高UA的溶解度也不断升高;但5 000和6 000μmol/L样本溶液中,在0 h时UA的溶解度随pH值的升高而升高,在24 h时UA的溶解度随pH值的升高呈现先升高后降低的趋势,当pH值为7.0时UA的溶解度最高,且测得浓度与理论浓度基本一致。(2)在PBS溶液中,0 h时对1 000、2 000、3 000、4 000、5 000和6 000μmol/L样本溶液中UA的溶解度进行检测,结果显示随pH值的升高UA的溶解度不断升高,当pH值为7.0及以上时,UA的溶解最高,且测得浓度与理论浓度基本一致;在24 h时对上述溶液进行重复检测,结果显示在pH值为8.0时,各样本溶液UA溶解度的测得浓度与理论浓度基本一致,当pH值降低时,UA的溶解度不断降低。在1 000、2 000和3 000μmol/L样本溶液中,当pH值为6.0和7.5时,UA的溶解度均出现大幅度下降的现象。在NaOH和PBS溶液中,除1 000μmol/L样本溶液外,其它样本溶液均出现不同程度的晶体析出,经过红外光谱测定晶体为UA。结论:pH值可以显著影响UA的溶解与析出。     

压电石英晶体传感器检测血浆纤维蛋白原方法的研究及应用

目的建立一种压电石英晶体传感器检测血浆纤维蛋白原(Fib)的方法.方法采用AT切向、基频10MHz的银膜石英晶体作为压电元件;观察血浆凝集过程中晶体振荡频率的变化,确定血浆凝集反应的起点和终点.结果 (1)检测池加入凝血酶原待频率稳定后,血浆标本加入反应体系后,晶体振荡快速下降,由于反应中形成的纤维蛋白原凝块的质量、体积与晶体接触的面积以及血清产生的量不同,造成频率瞬时的上升、下降,或频率稳定,直至反应结束频率平稳,加样后频率下降的起点为血浆凝集反应的起点,频率上升、下降,稳定的起始点为反应的终点,计算两点的时间即为反应时间,查标准曲线得纤维蛋白原的含量.(2)该方法的检测结果与STAGO磁珠检测法的相关性好(r＝0.957).结论压电石英晶体传感器检测血浆纤维蛋白原是一种实时在线检测的方法,在线观测凝集反应的终点,反应的起点和终点的判别方便、可靠、准确.且操作简单,快速,成本低廉,是一种敏感性高、重复性好的方法,并可用于临床检测.     

压电石英晶体传感器检测血浆凝血酶时间的实验研究

目的建立一种检测血浆凝血酶时间（TT）的压电石英晶体传感器。方法采用AT切向、基频10MHz的银膜石英晶体作为压电元件;探讨反应体系密度和黏度与晶体振荡频率变化的关系,观察血浆凝集过程中晶体振荡频率的变化规律,确定判断血浆凝集反应起点和终点的方法。结果（1）反应体系加入凝血酶试剂启动凝血反应后,晶体振荡频率呈阶梯状快速下降,接着频率瞬时上升,然后再缓慢降低达到平稳,频率阶梯下降的起点为血浆凝集反应的起始点,频率从快速上升变为下降的转折点为血浆凝集的终点,计算两点之间的时间即为血浆凝血酶时间。（2）该方法的检测结果与STAGO血凝仪磁珠检测法的相关性好（γ=0．964）,批内CV=2．26％,批间CV=7．53％。结论压电石英晶体传感器检测血浆凝血酶时间,血浆凝集反应起点和终点确定方便、可靠。且操作简单、快速、成本低廉,检测结果准确,是一种敏感性高、重复性好的血浆凝血酶时间检测新方法,可用于临床栓测。     

压电石英晶体振荡频率检测平台的建立及应用

目的　构建一种 2× 5型压电石英微阵列振荡频率检测平台。方法　利用基频 1 0MHz的 2× 5型石英晶体微阵列作为传感换能器件 ,自行设计自激式振荡频率检测仪、频率分析软件PESA2 .0 ,与计算机连接组成压电石英晶体传感器检测平台 ,并应用于气、液相检测。结果　石英晶体微阵列在气、液相起振能力强 ,并获得稳定振荡 ,气相稳定度可达± 1Hz ,液相≤ 2Hz。消除了温度、电离辐射、机械震动等因素的干扰 ;实现了频率信号检测的自动化、微机化和自能化。结论　该检测平台具有检测频率稳定、自动化和智能化程度较高、能实时检测等优点 ,是一种新型压电晶体振荡频率检测共用平台 ,可广泛用于压电传感器振荡频率的检测     

离子浓度对肽核酸压电基因传感器杂交效应的影响

目的　探索不同离子浓度的PBS缓冲液对肽核酸压电基因传感器阵列杂交效应的影响。方法　压电基因传感器阵列表面固定上 1 .5 μM的bis PNA探针后 ,分别观察在 1 0、2 0、5 0、1 0 0mM的PBS缓冲液环境中与相应的靶序列杂交 ,记录频率的下降值及反应所需的时间。结果　钠离子浓度从 1 0mM增加至 2 0mM时 ,杂交导致的频率下降值及平衡时间均增加 (P <0 .0 5 ) ;从 2 0mM增加至 5 0、1 0 0mM时 ,杂交导致的频率下降值各组间无显著差异 ,但杂交平衡时间却呈倍数增长。结论　离子浓度极大地影响了杂交反应的时间 ,低盐离子浓度更有利于提高bis PNA和dsDNA的杂交速率 ,高盐离子浓度则会大大降低bis PNA的结合速率。     

离子强度对压电传感器基因杂交动力学的影响

目的：探讨盐离子浓度对压电传感器基因杂交动力学的影响。方法：设计并合成金黄色葡萄球菌耐药基因探针及与其完全互补的靶序列，在石英谐振压电传感器阵列上进行杂交，杂交缓冲液中含5种NaCl浓度，计算机采集并分析传感器频率数据得出杂交动力学参数，同时比较各盐离子浓度下杂交达到平衡所需时间，杂交前后的频率差，平衡后频率波动幅度大小。结果：随NaCl浓度增加，结合速率常数，杂交平衡常数，最大杂交量均呈现明显增大趋势。而离解速率常数呈下降趋势，杂交前与杂交平衡后的频率差值增大，同时，杂交平衡后频率波动幅度相应增大，杂交达平衡所需时间延长。结论：一定的盐离子浓度促进基因杂交，但盐离子浓度过高则对晶体振荡性能产生负面影响。     

固定多黏菌素B的压电生物传感器检测内毒素

目的本研究拟探讨快速、定量和灵敏的内毒素(ET)检测方法。方法通过硅烷化的晶体表面将多黏菌素B(PMB)固定到ET切割10MHz压电石英晶体表面,研制成一种新型的用于检测血浆中ET浓度的压电生物传感器。结果该传感器检测中,ET在0.5～20.0μg/mL浓度范围内有良好的线性关系。结论该研究探讨了PMB浓度、溶液的离子强度和pH值以及反应时间等对ET吸附性能的影响,检测结果表明该传感器可用于临床和环境检测。     

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.