蝎毒抗癌多肽组分Ⅲ的分离方法研究

目的：探索蝎毒抗癌多肽组分Ⅲ(antineoplastic polypeptide-Ⅲfrom APBMV,AP-Ⅲ)的分离和纯化方法。方法：蝎毒经预处理后,分别采用pharose FF阳离子交换凝胶柱(cm×5 cm交换柱,流速0．8 ml／min,梯度洗脱1 200 min)层析法和Sepharose FF阳离子交换凝胶柱与Sephadex G-50凝胶柱(柱：100cm×5 cm;流速：0．5 ml／min)联合层析法,分离AP-Ⅲ,用HPLC仪检测2种方法分离出的AP-Ⅲ的纯度及产率。结果：单独用Sepharose FF阳离子交换凝胶柱分离,AP-Ⅲ产率为2．24％,纯度为89％。Sephadex G-50凝胶柱与Sepharose FF阳离子交换凝胶柱联用,其产率及纯度分别提高至4．72％和％。结论：Sephadex G-50与Sepharose FF联用可提高AP-Ⅲ的产率及纯度,从而为开发和利用AP-Ⅲ提供了实验依据。     

Searose FF凝胶柱分离参数变化对蝎毒抗癌多肽纯化的影响

目的优选蝎毒抗癌多肽(antineoplasticpolypeptidefromButhusmartensiivenom,APBMV)组分Ⅲ(antineoplas-cpolypeptide-ⅢfromAPBMV,AP-Ⅲ)的分离参数。方法采用不同长度的SepharoseFF阳离子交换凝胶柱及不同的样品洗脱速度,比较AP-Ⅲ的分离效果。结果当柱长为em,直径为5cm,流速为1．0ml／min时,可分离出6个峰,AP-Ⅲ纯度为89％;分别以0．8,0．5和0．2ml／min流速进行层析,可分别分离出6,4,3个峰,AP-Ⅲ相应的纯度为89％,85％,83％。当柱长为12．5cm,而其他条件相同时,均得到2个峰,相应的AP-Ⅲ纯度分别为83％,82％,％和78％。结论分离、纯化AP-Ⅲ较理想参数为柱长25cm,直径5em,流速0．8ml／min,梯度洗脱时间1200min。     

Sepharose FF凝胶柱分离参数变化对蝎毒抗癌多肽纯化的影响

目的 :优选蝎毒抗癌多肽 (antineoplasticpolypeptidefromButhusmartensiivenom ,APBMV)组分Ⅲ (antineoplas ticpolypeptide ⅢfromAPBMV ,AP Ⅲ )的分离参数。方法 :采用不同长度的SepharoseFF阳离子交换凝胶柱及不同的样品洗脱速度 ,比较AP Ⅲ的分离效果。结果 :当柱长为 2 5cm ,直径为 5cm ,流速为 1 .0ml/min时 ,可分离出 6个峰 ,AP Ⅲ纯度为 89% ;分别以 0 .8,0 .5和 0 .2ml/min流速进行层析 ,可分别分离出 6 ,4,3个峰 ,AP Ⅲ相应的纯度为89% ,85 %、83%。当柱长为 1 2 .5cm ,而其他条件相同时 ,均得到 2个峰 ,相应的AP Ⅲ纯度分别为 83% ,82 % ,80 %和 78%。结论 :分离、纯化AP Ⅲ较理想参数为柱长 2 5cm ,直径 5cm ,流速 0 .8ml/min ,梯度洗脱时间 1 2 0 0min。     

蝎毒抗癌多肽组分Ⅲ的分离方法研究

目的：探索蝎毒抗癌多肽组分Ⅲ的分离和纯化方法：方法：蝎毒经预处理后,分别采用ＳｅｐｈａｒｏｓｅＦＦ阳离子交换凝胶柱层析法和ＳｅｐｈａｒｏｓｅＦＦ阳离子交换凝胶柱ｅｐｈａｄｅｘＧ－５０凝胶柱联合层析法,分离ＡＰ－Ⅲ,用ＨＰＬＣ仪检测２种方法分离出的ＡＰ－Ⅲ的纯度及产率。结果：单独用ｅｐｈａｒｏｓｅＦＦ阳离了交换凝胶柱分离,ＡＰ－Ⅲ产率为２．２４％,纯度为８９％。ＳｅｐｈａｄｅｘＧ－５０凝胶柱与Ｓｅｐ     

蝎毒抗癌多肽及其纯化组分Ⅰ及Ⅲ的毒性观察

目的 :观察蝎毒抗癌多肽 (APBMV)纯化组分Ⅰ (AP Ⅰ )及Ⅲ (AP Ⅲ )与蝎毒抗癌多肽之间的毒性差别 ,为前二者药效学研究提供新的实验依据。方法 :取昆明种小鼠 180只 ,随机分为 18组 ,每组 10只 ,腹腔一次注射0 .2ml/ 2 0g不同浓度的APBMV(1～ 6组 )、AP Ⅰ (7～ 12组 )及AP Ⅲ (13～ 18组 ) ,观察动物的行为学改变 ,并取出死亡动物脏器进行病理学观察 ,按照综合计算法得出AP Ⅰ、AP Ⅲ与APBMV的LD50 和 95 %可信区间。结果 :小鼠注射高浓度的APBMV、AP Ⅰ或AP Ⅲ后 ,在 2min后即出现明显的烦躁、尖叫、嘶咬、多涎 ,继而呆滞、呼吸不规则、颈毛直立等行为学改变 ,随着给药时间的延长 ,动物会表现行动迟缓、竖尾、腹泻、喘息性呼吸 ,后肢强直 ,窒息直至死亡。而其他各组在 3min后才出现程度不同的毒性反应。高浓度组动物在 2 5min内全部死亡 ,其他各组部分动物在 6 0min内相继死亡。死亡和处死动物内脏无明显病理学改变。APBMV、AP Ⅰ或AP Ⅲ的LD50 和 95 %可信区间分别为 1.386 ,1.118,1.145和 (1.386± 0 .2 47) ,(1.118± 0 .194) ,(1.145± 2 0 .198)。结论 :APBMV经进一步纯化后 ,其有效成分AP Ⅰ及AP Ⅲ毒性均有显著提高     

蝎毒抗癌多肽及其纯化组分Ⅰ及Ⅲ的毒性观察

目的观察蝎毒抗癌多肽(APBMV)纯化组分Ⅰ(AP-Ⅰ)及Ⅲ(AP-Ⅲ)与蝎毒抗癌多肽之间的毒性差别,为前二者药效学研究提供新的实验依据.方法取昆明种小鼠180只,随机分为18组,每组10只,腹腔一次注射0.2ml/20g不同浓度的APBMV(1～6组)、AP-Ⅰ(7～12组)及AP-Ⅲ(13～18组),观察动物的行为学改变,并取出死亡动物脏器进行病理学观察,按照综合计算法得出AP-Ⅰ、AP-Ⅲ与APBMV的LD50和95%可信区间.结果小鼠注射高浓度的APBMV、AP-Ⅰ或AP-Ⅲ后,在2min后即出现明显的烦躁、尖叫、嘶咬、多涎,继而呆滞、呼吸不规则、颈毛直立等行为学改变,随着给药时间的延长,动物会表现行动迟缓、竖尾、腹泻、喘息性呼吸,后肢强直,窒息直至死亡.而其他各组在3min后才出现程度不同的毒性反应.高浓度组动物在25min内全部死亡,其他各组部分动物在60min内相继死亡.死亡和处死动物内脏无明显病理学改变.APBMV、AP-Ⅰ或AP-Ⅲ的LD50和95%可信区间分别为1.386,1.118,1.145和(1.386±0.247),(1.118±0.194),(1.145±20.198).结论[HTSSAPBMV经进一步纯化后,其有效成分AP-Ⅰ及AP-Ⅲ毒性均有显著提高.     

高效毛细管电泳及其在体内药物分析中的应用

高效毛细管电泳(HighPerformanceCapillary Electrophoresis,HPCE)是近年来发展较快的一种分离、分析技术,是经典电泳技术和现代微柱分离相结合的产物。HPCE和高效液相色谱法(HPLC)相比,相同处在于都是高效分离技术,仪器操作均可自动化,且均有多种不同分离模式。差异在于,HPCE用迁移时间取代HPLC中的保留时间,分析时间通常不超过30min,比HPLC速度快;对HPCE而言,从理论上推得其理论塔板高度和溶质扩散系数成正比,对扩散系数小的生物大分子而言,其柱效就要比HPLC高得多;HPCE所需样品为nl级,最低可达270fl,流动相用量也只需几…     

SepharoseFF凝胶柱分离参数变化对蝎毒抗癌多肽纯化的影响

目的：优选毒抗癌多肽组分Ⅲ的分离参数,方法：Ｅ 同长度的ＳｅｐｈａｒｏｓｅＦＦ阳离子交换凝胶柱及不同样品洗脱速度,比较ＡＰ－Ⅲ的分离效果。结果：当柱长为２５ｃｍ,直径为５ｃｍ,流速为１．０ｍｌ／ｍｉｎ时,可分离,ＡＰ－Ⅲ纯度为８９％;分别０．８,０．５和０．２ｍｌ／ｍｉｎ流速进行层析,可分别同６,４,３个峰,ＡＰ－Ⅲ相应的纯度为８９％,８５％,８３％。当柱长为１２．５ｃｍ,而其他条件相同时,得到     

东亚钳蝎蝎毒的指纹图谱

目的 :探索东亚钳蝎蝎毒 (scorpionofButhusmartensiivenom)的分析及鉴定方法 ,建立相关质量标准。方法 :采用高效液相色谱法 (highperformanceliquidchromatography ,HPLC)分析检测 8个蝎毒样品。结果 :蝎毒可分离出40多个组分 ,实验所使用的 8个省内所产蝎毒具有非常相似的图谱。蝎淋巴液和 3种蛇毒与蝎毒图谱有显著不同。同一样品 (二次冻干粉 )连续测定 5次进样量的精密度、重现性均达到有关规定。结论 :采用HPLC法获得了蝎毒的指纹图谱 ,并测定了可影响蝎毒质量的蝎淋巴液和可能用于掺伪的 3种蛇毒的图谱。本研究为蝎毒及其制剂的进一步开发应用提供了有价值的实验依据。     

蝎毒促增殖肽在大鼠骨髓间充质干细胞上靶蛋白的初步研究

目的研究蝎毒促增殖肽Bmkpp在大鼠骨髓间充质干细胞(BMSCs)中的靶点。方法采用Sepha-dex G-50凝胶层析和CM-Sepharose Fast Flow弱阳离子交换层析从东亚钳蝎蝎毒中分离纯化出蝎毒促增殖肽Bmkpp,RP-HPLC检测,酶解-质谱鉴定;提取大鼠BMSCs全蛋白、膜蛋白、胞浆蛋白后上样到Bmkpp亲和层析柱,洗脱收集靶蛋白峰,超滤离心法脱盐浓缩;SDS-PAGE电泳分析洗脱蛋白。结果分离纯化出Bmkpp和其相邻的峰4,RP-HPLC检测分离纯化的Bmkpp纯度达95%。酶解质谱法鉴定其与Bmkpp标准品的主要成分一致。一步亲和层析洗脱峰均为2个,电泳结果显示胞浆洗脱蛋白条带较多,但细胞膜无结合蛋白条带。改变盐浓度分步梯度洗脱全蛋白共得4个峰,1 mol/L NaCl盐浓度分步洗脱全蛋白于13、14、40、260 kD处有4条较明显的电泳条带。结论 Bmkpp在BMSCs上作用于多个靶点并有其特异性的靶标。     

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.     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

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.     

CLINICAL SIGNIFICANCE OF SERUM CYTOKINES IL-1β,sIL-2R,IL-6,TNF-α,AND IFN-v IN ACUTE CORONARY SYNDROME

Objectives To explore serum cytokines levels (including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v) and their significance in patients with acute coronary syndrome (ACS) and the subsequent follow-ups, with attempt to estimate the role of various serum inflammatory markers in the diagnosis and assessment of ACS.Methods The study population include 40 patients with acute myocardial infarction (AMI), 40 patients with unstable angina pectoris (UAP), and 40 controls. Among the 80 patients, 60 patients attended a follow up 4 months later. Serum inflammatory markers including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v were measured by enzyme linked immunosorbent assay.Results Serum IL- 1 β, sIL-2R, IL-6, TNF-α were significantly higher in AMI group or UAP group compared to the control group and became significantly lower 4 months later in the follow-up patients. Serum levels of IFN-v shows no significant difference between AMI group or UAP group and controls, also showing no significant change when measured in follow up patients. There was no correlation between serum creatine kinase-MB isoenzyme levels and serum inflammatory markers either in UAP or AMI group. Furthermore, when divided into two subgroups using Wagner's QRS scoring system in the AMI group, there is no difference of each serum inflammatory marker between ≤ 6 scores group and ＞ 6 scores group.Conclusion Serum levels of certain inflammatory markers may have some diagnostic value for ACS, and can be a useful marker reflecting disease stability.