壳聚糖固定α-淀粉酶的研究

目的　从虾蛄壳提取壳聚糖 ,将α-淀粉酶固定在壳聚糖上。　方法　以自制壳聚糖为载体 ,一氯乙酸或三氯乙酸为交联剂 ,将α-淀粉酶固定化。　结果　三氯乙酸是较好的交联剂。固定化酶与原酶的最适 p H及最适温度基本相同 ,均为 6.5 ,5 0℃ ;固定化酶在 65℃仍有较高热稳定性 ,而原酶活力明显下降 ;固定化酶的表观米氏常数 Km =0 .6× 10 -2 g/ml,原酶 Km =1.1× 10 -2 g/ml;一定浓度 Cl-、Br-离子有助提高固定化酶活力。　结论　该固定化酶活力较高 ,热稳定性和贮存稳定性好 ,是良好的检测试剂。     

固定无花果蛋白酶制备可溶性肽的研究

将 Sephadex G-200与β-硫酸酯乙砜基苯胺(SESA)首先醚化制备对氨基苯砜乙基交联葡聚糖(ABSE-Sephadex G-200),然后经重氮化固定无花果蛋白酶。固定化酶的活力回收达69%。苯甲酰-DL-精氨酰-β-蓁胺(BANA)对该酶固定化过程中的活性变化有保护作用。天然酶与固定化酶都具有良好的耐热性,在59～60℃,80min,活性均无明显下降;在69～70℃,80min 固定化酶较天然酶更稳定。用 BANA 为底物,在半胱氨酸存在下,测定了两种形式酶的动力学性质。在 pH7.7的磷酸盐缓冲系统中,37℃天然无花果蛋白酶的 Km=0.32m-mol/L;在间歇振摇下固定化酶的表观 Km′=1.02mmol/L。最适 pH 无明显改变,均为7.7。     

甲壳胺-羧甲基纤维素钠高分子复合物固定酶的研究

本文采用天然高分子甲壳胺与羟甲基纤维素钠的高分子复合物作哉体,固定葡萄糖氧化酶和纤维素酶。得到的固定化酶活性较高、热稳定性好。固定化后的葡萄糖氧化酶比固定化前的最适反应温度提高50℃,固定化后的纤维素酶最适反应温度提高20℃。固定酶的贮存稳定性明显提高,并可反复使用。     

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。     

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.     

固定化脂肪酶的动力学研究

以壳聚糖为载体，用物理吸附固定化脂肪酶，对影响固定化过程的各种因素进行了考察，确定了最优条件，并比较了游离酶和固定化酶的最适温度、pH、酶的热稳定性以及表观Km。固定化酶水解的最适pH(pH7．0)稍低于游离酶(pH8．0)，最适温度为50℃(游离酶为40℃)。固定化酶在50℃的半衰期(0．96h)是游离酶(0．32h)的3倍，在4℃的冰箱中贮存3个月活力变化很小。     

聚甲基丙烯酸缩水甘油酯固定化青霉素酰化酶性质的研究

应用反相悬浮技术合成了珠状甲基丙烯酸缩水甘油酯 - N ,N′-亚甲基双 (丙烯酰胺 )共聚物 ,并将巨大芽孢杆菌 ( Bacillus megaterium)青霉素酰化酶共价偶联到甲基丙烯酸缩水甘油酯共聚物载体上 ,制成固定化青霉素酰化酶 ,其表观活性为 3 71 U/g(干重 ) ,水解青霉素 G钾盐的最适温度为 47°C,最适 p H为 8.5 ,在 p H4.5～ 9.0 ,温度 40°C以下时酶的活性稳定 ,表观米氏常数 Km为 1 .3 3× 1 0 -2 mol/L ,最大反应速度 vmax为 1 .2 7× 1 0 -5mol/min,固定化酶在 4°C冰箱保存 3 5 d,再水解 w=0 .0 2的青霉素 G钾盐溶液 ,重复使用 3 0次 ,保留酶活性 88.1 %。     

壳聚糖固定胰蛋白酶的研究

目的：从蟹壳中提取壳聚糖，将胰蛋白酶固定在从蟹壳中提取的壳聚糖上。方法：以自制壳聚糖为载体，戊二醛为双官能团交联剂，将胰蛋白酶固定化。结果：０．２ｇ壳聚糖（干）与４％戊二醛交联后再与５．０ｍｇ胰蛋白酶固定结合，效果较好。固定化酶的表现米氏常数Ｋ′ｍ＝１．５ｍｍｏｌ／Ｌ，而天然酶Ｋｍ＝７．６ｍｍｏｌ／Ｌ；固定化酶最适温度为８０℃，比天然酶提高了２０℃；固定化酶最适ｐＨ为７．０．而天然酶为８．０。该载体来源广，易提取，固定化酶的贮存稳定性很好，２个多月重复使用，酶活力未见明显下降     

蒜酶催化蒜氨酸动力学研究

目的:研究酶促反应的动力学参数,为新药设计提供依据。方法:采用HPLC方法测定酶促反应的动力学参数及蒜酶的最适pH和最适温度。结果:蒜酶以蒜氮酸为底物,最大反应速度Vmax=27.9 μnml·min-1·mg-1,米氏常数Km=3.6 mmol/L,其最适pH为6.6,最适温度为35℃。结论:酶促反应的动力学参数为蒜酶寻求有利的反应条件、更大限度地提高酶反应的效率提供了参考数据。     

蕲蛇酶(凝血酶样酶)溶液及注射液的稳定性研究

+本文报道温度及pH对蕲蛇酶(凝血酶样酶)溶液及注射液活性的影响。经50,60,80,100±0.5℃恒温水浴中温孵1h后测定凝血酶活力及pH的变化,以判定蕲蛇酶的稳定性。结果50℃热处理酶活力未变,60℃以上酶活力逐渐降低,80,100℃酶活力减少50％～78％。热处理后对pH影响不大。以1:nol/L HCl或Dnol/L NaOH调整蕲蛇注射液的pH至1～12,分别测定凝血酶活力,共最适pH为6,pH在5～8范围内酶活力较稳定,在此范围以外酶活力均受影响同。当调回到pH6时,除ph1,12外,仍表现出很强的酶活力。     

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.     

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.     

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.