DNA引物酶的提取分离与引物合成活性的研究

以小鼠艾氏腹水癌细胞经超声破碎的提取液为材料，依次用ＤＥ－５２和Ｐ－１１层析柱进行离子线的洗脱，分别在ＫＣｌ浓度的０．１７～０．２ｍｏｌ／Ｌ和０．２５～０．２７ｍｏｌ／ｌ处ＤＮＡ引物酶被洗脱下来。用大肠杆菌大片段ＤＮＡ多聚酶Ｉ延长引物法检测Ｄ引物酶比活性为８７５３Ｕ／ｍｇ，酶总获得为２２．１％，放射自显影法检测引物合成活性显示引物酶合成了不同长度的引物。     

聚苯乙烯载体亲和层析纯化尿激酶

聚苯乙烯经磺化后连接手臂６－氨基己酸，然后偶联配基对氨基苯甲脒，制成亲水性的亲和载体。载体的最适配基密度为每克湿载体３２μｍｏｌ，最适吸附条件为ｐＨ７．５、０．０５ｍｏｌ／Ｌ磷酸缓冲液（含ＮａＣｌ０．５ｍｏｌ／Ｌ）。在ｐＨ４．０、０．１ｍｏｌ／Ｌ醋酸缓冲溶液（含ＮａＣｌ０．５ｍｏｌ／Ｌ）中，采用０～１ｍｏｌ／Ｌ　ＮａＳＣＮ梯度洗脱，可将比活８．３３μｍｏｌ·Ｓ－１·ｍｇ－１的粗酶纯化５０倍，收率大于５３％。     

马桑内酯对脑γ－氨基丁酸分泌和谷氨酸脱羧酶及其受体结合力的影响

为探讨癫痫发病的生物化学机制，用培养的大鼠大脑皮层神经元和大脑皮层匀浆及突触膜为模型，采用酶联免疫法，放射分析法观察马桑内酯（ＣＬ）对大鼠脑内γ－氨基丁酸（ＧＡＢＡ）分泌，谷氨酸脱梭酶（ＧＡＤ）活性和谷氨酸（Ｇｌｕ）受体结合力的影响。结果表明，５０μｍｏｌ／ＬＣＬ可抑制ＧＡＢＡ分泌，１２、２４、４８小时抑制率为８．３％～１４。５％；在０．１５～１５０μｍｏｌ／Ｌ范围，ＣＬ可抑制ＧＡＤ活性，抑制率为１．３２％～１８．７６％；ＣＬ在２．８～３５０μｍｏｌ／Ｌ范围内可降低Ｇｌｕ受体结合力，降低程度为１８．８％～３１．６％，且各剂量组与对照组差异均有极显著意义（Ｐ＜０．０１）。证实ＣＬ可抑制培养神经元ＧＡＢＡ分泌，并呈时间依赖和剂量依赖关系。     

β—L—2‘,3’—双脱氧—5—氟胞苷在体外对乙型肝炎病毒复制的抑制作用

目的：研究β－Ｌ－２＇，３＇－双脱氧－５－氟胞苷的抗乙型肝炎病毒作用。方法：从２．２．１５细胞培养液中提取乙肝病毒ＤＮＡ，用ＤＮＡ印迹法分别药物对乙肝病毒ＤＮＡ复制的影响。观察药物对人Ｔ－成淋巴样细胞的生长抑制作用，用狭线印迹法检测人Ｔ－成淋巴样细胞的线粒体ＤＮＡ。结果：β－Ｌ－２＇，３＇－双脱氧－５－氟胞苷对乙肝病毒ＤＮＡ合成的半数抑制浓度为０．０５μｍｏｌ／Ｌ，但其抗乙肝病毒作用是可逆的。对人     

血清总胆汁酸双试剂连续监测的临床意义

目的：利用全自动生化分析仪，建立一种快速检测血清总胆汁酸（ＴＢＡ）活性的方法和正常参考值。方法：双试剂酶偶联连续监测法，检测６０例不同肝病患者和３００名健康成人血清ＴＢＡ活性。结果：其测值（ｘ±ｓ）分别为４８．９５±５．７３μｍｏｌ／Ｌ和５．１２±３．４６μｍｏｌ／Ｌ，差异有显著性（Ｐ＜０．０１）；本法批内ＣＶ分别为１．９９％、２．８５％和３．００％，批间ＣＶ分别为３．２８％、３．４０％和３．１６％，线性范围０～２００μｍｏｌ／Ｌ。结论：双试剂酶偶联连续监测法，线性好，特异性强，灵敏度高，快速、准确，对肝脏疾病的诊断及临床分级具有重要价值。     

转染bcl-2基因抑制足叶乙甙诱导的K562细胞凋亡

目的：观察ｂｃｌ２对Ｋ５６２细胞凋亡的影响。方法：用真核表达质粒ｐ２９０ｂｃｌ２转染Ｋ５６２细胞，用ＲＴＰＣＲ和Ｗｅｓｔｅｒｎｂｌｏｔ检测ｂｃｌ２ｍＲＮＡ和蛋白表达；用ＭＴＴ实验检测Ｋ５６２细胞对化疗药物足叶乙甙的敏感性变化；用ＤＮＡ“ｌａｄｄｅｒ”检测ｂｃｌ２对凋亡的影响。结果：转染ｂｃｌ２后Ｋ５６２的ｂｃｌ２蛋白表达增高，细胞存活率明显提高，在足叶乙甙低于１２５μｍｏｌ／Ｌ时检测不到凋亡梯形ＤＮＡ；而Ｋ５６２细胞在足叶乙甙为３２μｍｏｌ／Ｌ时就能检测到凋亡梯形ＤＮＡ。结论：若把ｂｃｌ２导入正常造血干细胞，从而增强干细胞对化疗药物的耐受性，这将对化疗具有重要意义。     

丙酮酸脱羧酶催化合成L-苯基乙酰基甲醇的研究

用酵母的丙酮酸脱羧酶粗提物催化苯甲醛合成Ｌ－苯基乙酰基甲醇（Ｌ－ＰＡＣ）,后者为合成Ｌ－麻黄素的前体。酶转化反应的最佳温度为１０℃,ｐＨ６．８,时间为５～６ｈ,酶用量７ｕ／ｍｌ,乙醇２．０ｍｏｌ／Ｌ,苯甲醛１５０～１８０ｍｍｏｌ／Ｌ,丙酮酸１５０～２００ｍｍｏｌ／Ｌ,转化液中Ｌ－ＰＡＣ浓度可达１４０ｍｍｏｌ／Ｌ。     

肾上腺素能受体激动剂与阻断剂对血管平滑肌细胞增殖和c－fos，c－myc原癌基因表达的影响

目的：研究肾上腺素能受体激动剂与阻断剂对血管平滑肌细胞增殖、ＤＮＡ合成和ｃ－ｆｏｓ，ｃ－ｍｙｃ原癌基因表达的影响．方法：应用细胞培养、￣３Ｈ－胸腺嘧啶核苷掺入和斑点杂交的方法．结果：１～１０μｍｏｌ／Ｌ去甲肾上腺素（ＮＥ）和０．１～１０μｍｏｌ／Ｌ异丙基肾上腺素（ＩＳＯ）可明显促进离体培养的ＳＤ大鼠的主动脉平滑肌细胞增殖、￣３Ｈ－ＴｄＲ掺入和ｃ－ｆｏｓ，ｃ－ｍｙｃ原癌基因表达，并呈剂量依赖效应．该效应可为相应的受体阻断剂Ｐｈｅｎｔｏｌａｍｉｎｅ（１０μｍｏｌ／Ｌ）和Ｐｒｏｐｒａｎｏｌｏｌ（１０μｍｏｌ／Ｌ）所抑制（Ｐ＜０．０５）．结论：肾上腺素能受体激动剂可促进离体培养的大鼠主动脉平滑肌细胞的增殖、ＤＮＡ合成和原癌基因的表达，该效应可被其相应的阻断剂所抑制．     

用固定化德阿昆合假单孢生产L－丙氨酸

用固定化德阿昆合假单孢生产Ｌ－丙氨酸刘景晶，吕伟峰，吴梧桐．药物生物技术，１９９５，２（１）：４０～４４用卡拉胶固定含有Ｌ－天门冬氨酶－β脱羧酶活性的德阿昆合假单孢。该固定化细胞经０．１ｍｏｌ／Ｌ天冬氨酸铵，０．１ｍｍｏｌ／ＬＰＬＰ和０．２ｍｏｌ／Ｌ...     

银川市学龄前儿童尿铅“建议限值”的探讨

为了调查儿童体内尿铅含量的“建议限值”，作者于１９９５年９～１２月，选择银川市新市区、新城、老城区３～６岁学龄前儿童并选其父母均不从事铅作业者、居住地远离铅工矿区、在银川市居住３年以上的健康儿童为检测对象。随机抽样２８４人，其中男１５６人，女１２８人，采集空腹晨尿，用双硫腙法进行含铅量的测定。结果：男童ｘ为０．１５９μｍｏｌ／Ｌ，女童ｘ为０．１７１μｍｏｌ／Ｌ，经ｕ检验男女尿铅含量之间无显著性差异。故学龄前儿童铅含量可用同一“建议限值”来表示。经计算，３～６岁儿童尿铅ｘ为０．１６５μｍｏｌ／Ｌ，标准差为０．０７９６μｍｏｌ／Ｌ，故９５％的参考值上限为０．３７５μｍｏｌ／Ｌ。作者建议将尿铅含量０．３７５μｍｏｌ／Ｌ作为学龄前儿童尿铅水平的“建议限值”，并列入儿童正常体检指标。从而为进一步了解和控制铅污染、提高和保护儿童健康水平提供理论依据     

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.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

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.     

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.