甲壳糖固定血管紧张素转换酶的制备及其性质

目的 将血管紧张素转换酶(ACE)固定化并测定其动力学性质。方法 以甲壳糖为载体，碳二亚胺(EDC)为交联剂，与分离纯化的ACE混合,使ACE固定化；并以马尿酰甘氯酰甘氨酸(HCG)为底物，分别测定了其游离酶和固定化酶的动力学性质。结果 固定化酶活力110u／g湿重，固定化率为22％，与游离酶比较，固定化酶稳定且具良好的耐热性，在50—80℃固定化酶较游离酶稳定。在pH7．1的底物缓冲体系中，37℃，游离酶Km=0.026mol／L；在间歇振摇下固定化酶的表现Km=0．040mol／L。游离酶最适pH为、8.0固定化酶最适pH为9．0。结论 用甲壳糖固定ACE方法简便可行，且固定后的甲壳糖性能稳定，有广阔的应用前景。     

PGA在含环氧活性基的多孔高聚物载体上的固定化及修饰

研究了青霉素G酰化酶(PGA)在含环氧活性基的多孔高聚物载体上的固定化及修饰,优化固定化条件为1 mol/L,pH 8.0的磷酸钾缓冲体系,每克载体(湿重)投酶量为500~550 U,30℃下150 r/min固定化36~48 h,得到的固定化酶表观酶活为每克载体(湿重)177 U,表观酶活回收率35%。固定化酶经巯基乙醇修饰后提高了热稳定性。固定化酶水解青霉素G的最适pH为9.0,最适温度为47℃,在pH 4~9,40℃以下稳定。固定化酶的各项性能均优于游离酶。     

可溶性固定化木瓜蛋白酶的制备及其性质研究

目的 制备一种可溶性固定化木瓜蛋白酶并对其部分酶学性质进行研究.方法 将木瓜蛋白酶偶联在亲水性高分子聚合物羧甲基纤维素醋酸琥珀酸酯(AS-L)上制备得到水溶性固定化木瓜蛋白酶.在相同的条件下测定游离的木瓜蛋白酶和水溶性固定化木瓜蛋白酶的活力,得到两种状态的酶的最适作用pH值和最适作用温度,再于它们各自的最适条件下测定活力,得到各自的Km值;在相同的条件下测定两种状态的酶的热稳定性、酸碱稳定性、低温稳定性和回收稳定性.结果 游离木瓜蛋白酶和可溶性固定化木瓜蛋白酶的最适作用pH值分别为6.0和5.0,最适作用温度分别为60℃和70℃,Km值分别为2.53、3.07mg·ml-1;可溶性固定化木瓜蛋白酶在60℃条件下,保温12h,仍有62%的活性;在4℃条件下,30d后活性保留达90%以上;在pH6.0时稳定性最好,在pH4.0-7.0范围内也比较稳定(相对活性＞80%).结论 可溶性固定化木瓜蛋白酶能够在pH5.5以上的溶液中完全溶解,最适作用pH值范围变窄,最适作用温度和Km值升高;热稳定性和酸碱稳定性均明显增强.     

纳米二氧化硅固定木瓜蛋白酶性能研究

目的：研究固定化木瓜蛋白酶的性质。方法：以纳米二氧化硅为载体,戊二醛为交联剂,制备固定化木瓜蛋白酶。研究木瓜蛋白酶的最佳固定化条件以及固定化酶的酶学性质和动力学参数。结果：固定化酶的最适反应温度70℃,最适反应pH值为7.5,酶活回收率可达61.5％。结论：固定化酶催化的活性与游离酶相比不受显著影响。     

AS1-398中性蛋白酶固定化条件的初步研究

考察壳聚糖、卡拉胶、海藻酸钠、琼脂等固定化载体对ＡＳ１３９８中性蛋白酶固定化的影响。确定０３％戊二醛在３０℃,ｐＨ８．０的条件下处理壳聚糖８１０ｈ,加酶液,固定８ｈ,酶活力回收约为７７％,固定化酶最适作用温度为５５℃,最适作用ｐＨ为８０;固定化酶的稳定性比游离酶的稳定性有显著提高。     

色氨酸酶基因工程菌固定化及其培养条件的研究

目的 :对色氨酸酶基因工程菌ＷＷ 11进行固定化及培养条件研究 ,为工业化生产Ｌ 色氨酸奠定基础。方法 :通过色氨酸酶活力测定 ,考察三种固定化材料及温度、ｐＨ、单价阳离子降乙醇对固定化ＷＷ 11色氨酸酶活力的影响。结果 :以聚乙烯醇作为ＷＷ 11的固定化载体 ,其活力回收为 6 0 9%。固定化ＷＷ 1色氨酸酶降解反应最适 ｐＨ为 9 0、最适温度为 5 0℃ ;固定化ＷＷ 1色氨酸酶合成反应最适 ｐＨ为 7 5、最适温度为 45℃。Ｋ 、ＮＨ 4对固定化工程菌色氨酸酶有明显激活作用 ,而Ｎａ 则有一定的抑制作用。结论 :固定化工程菌色氨酸酶对温度…     

以甲壳胺为载体固定乙酰胆碱酯酶

目的 探讨以甲壳胺为载体固定乙酰胆碱酯酶的最佳条件及固定化酶的理化性质.方法 以戊二醛为交联剂,将乙酰胆碱酯酶固定在甲壳胺上,并分别测定固定化酶与溶液酶的活力.结果 固定化酶与溶液酶的最适温度相同,均为37℃;固定化酶与溶液酶最适pH分别为8.2、8.0;固定化酶在50℃仍有较高热稳定性,而溶液酶活力明显下降;固定化酶的米氏常数Km=0.84 mmol/L,溶液酶的Km=1.16 mmol/L;固定化酶活力回收为44.66%,在重复使用8次后仍能保持原有活力的50%.结论 该固定化酶与溶液酶相比,其热稳定性提高,与底物的亲和能力增大,且重复使用性较好.     

肽脱甲酰基酶在偶极离子修饰亲水磁珠表面的固定化及表征

目的 比较表面修饰偶极离子且带Ni2+-NTA或羧基的两种亲水磁珠对结核分枝杆菌肽脱甲酰基酶(Mycobacterium tuberculosis peptide deformylase,MtPDF)的固定化效果,探讨适于磁分离联用HPLC-MS筛选MtPDF配体混合物库的靶酶固定化方案.方法 将N端带有6xHis标签MtPDF的质粒pET-28a(+)导入E.coli BL21(DE3)重组表达,使用镍离子螯合亲和层析介质纯化后表征其酶学性质.将MtPDF与Ni2+-NTA或羧基磁珠偶联并比较其固定化容量、保留活性、固定化前后酶稳定性及对抑制剂放线酰胺素(Actinonin)的响应.结果 Ni2+-NTA磁珠固定MtPDF的容量为(99.7±4.5)mg/g(n = 3);最优投料比酶量∶磁珠量=1∶15时,固定化酶保留活性为(107.2±13.9)％(n = 3),在0℃、pH = 7.5缓冲液中振摇3 h活性仅保留43％.羧基磁珠活化后通过酰胺键共价固定MtPDF的容量为(36.9±2.6)mg/g(n=3);最优投料比酶量∶磁珠量=1∶60时,固定化酶保留活性为(102.6±15.4)％(n=3),与Ni2+-NTA磁珠固定化保留活性相当;在0℃、pH7.5缓冲液中振摇8 h活性无显著改变;羧基磁珠固定化酶对Actinonin的抑制响应与游离酶无显著差异(P＞0.05).结论 羧基磁珠固定化MtPDF酶活更稳定,不影响抑制剂响应,适于混合物库中MtPDF高亲和力配体筛选.     

α－淀粉酶在壳聚糖上的固定化研究

报道从蟹壳中提取壳聚糖的方法。以壳聚糖为载体，戊二醛为双官能团交联剂，将α－淀粉酶固定化。结果表明固定化α－淀粉酶与原酶的最适ｐＨ值基本相同，且均有２个最适ｐＨ值，分别为５和６．５；固定化酶的表观米氏常数ｋｍ′＝０．３３×１０￣（－２）ｍｇ／ｍｌ，而原酶ｋｍ＝１×１０￣（－２）ｍｇ／ｍｌ；固定化酶的热稳定性明显高于原酶，固定化酶的最适温度为６５℃，原酶为５０℃。     

基因工程菌CTB2的固定化及其制备L—苯丙氨酸的研究

对以卡拉胶为载体固定化基因工程菌ＣＴＢ２制备Ｌ－苯丙氨酸进行了研究。比较了固定化细胞的最适温度和最适ＰＨ，考察了该固定化细胞的柱以应特性以及产物Ｌ－苯丙氨酸的分离方法。结果表明，固定化细胞的最适温度为３０℃，最ＰＨ为７．０，柱反应最佳空时速度ＳＶ＝０．１９ｈ＾－１底转化率为２０％，固定化酶半衰期大于３０ｄ。     

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.