The Topical Use of Aprotinin in Cardiac Surgery with Cardiopulmonary Bypass

Objective To investigate the effects of the topical use of aprotinin on thebasis of comprehensive blood conservations in cardiopulmonary bypass (CPB). Methods In a prospectiveclinical trial, 20 patients were randomly divided into 2 groups. Control group: placebo was usedtopically. Aprotinin group: aprotinin was poured into the pericardial cavity before closure of thesternotomy. Before and 24h after surgery, hemoglobin (Hb), hematocrit (Hct), bleeding time (BT),clotting time (CT) and prothrombin time (PT) were measured. Meanwhile, amounts of the mediastinaldrainage and the hemoglobin loss were observed at 0, 2, 6 and 24h after operation. The samples fromthe mediastinal drainage were also collected to measure D-Dimer (D-D), tissue type plasminogenactivator (t-PA) activity, plasminogen activator inhibitor (PAI) activity and protein C (PC).Results In Aprotinin group, D-D, t-PA activity and PC were significantly reduced, compared withthose in Control group (P<0.05, P<0.05, P<0.01). On the contrary, PAI activity was significantlyincreased, compared with that in Control group. Amounts of the mediastinal drainage and thehemoglobin loss were decreased by 43% and 52%, compared with those in Control group. Conclusion Ourresults suggest that the topical use of aprotinin can have better effects on the basis ofcomprehensive moderate blood conservation.

Platelet activation and the protective effect of aprotinin in hepatolithiasis patients

OBJECTIVE: To explore platelet activation and the protective effect of aprotinin in patients with hepatolithiasis. METHODS: The count of plaletets and levels of CD_(62P) and CD_(63) were measured by flow cytometry in 38 patients with hepatolithiasis. Several measurements were carried out after treatment with aprotinin. RESULTS: The levels of CD_(62P), CD_(63) in patients with hepalolithiasis were higher than those in patients with cholecystolithiasis (P<0.05), but the count of platelets was lower (P<0.05). After operation, the levels of CD_(62P), CD_(63) were significantly increased in patients with hepatolithiasis, but the count of platelets was lower (P<0.05). Postoperative levels of CD_(62P), CD_(63) were significantly lower in patients treated with aprotinin than in normal controls (P<0.05); but there was no significant change in the count of platelets in the two groups. CONCLUSION: Platelet activation occurs in patients with hepatolithiasis, and may be inhibited by aprotinin.

Requirements for transfusion and postoperative outcomes in orthotopic liver transplantation:A meta-analysis on aprotinin

AIM:To study the effect of aprotinin used in orthotopic liver transplantation (OLT) on the intraoperative requirement for blood products and on the incidence of laparotomy for bleeding, thrombotic events and mortality. METHODS: A systematic review of the literature in the electronic database Medline and the Clinic Trials Registry Database was performed. Literature that did not fit our study were excluded. Patients in the reviewed studies were divided into two groups; one group used aprotinin (aprotinin group) while the other did not (control group). The data in the literature that fit our requirements were recorded. Weighted mean differences (WMD) in the requirements for blood products between the aprotinin group and the control group were tested using a fixed effect model. A Z test was performed to examine their reliability; the Fleiss method of fixed effect model was used to analyze data on postoperative events, and odds ratios (ORs) were tested and merged. RESULTS: Seven citations were examined in our study. Among them, a requirement for blood products was reported in 4 studies including 321 patients, while postoperative events were reported in 5 studies including 477 patients. The requirement for red blood cells and fresh frozen plasma in the aprotinin group was statistically lower than that in the control group (WMD=-1.80 units, 95% CI,-3.38 to-0.22; WMD=-3.99 units, 95% CI,-6.47 to-1.50, respectively). However, no significant difference was indicated in the incidence of laparotomy for bleeding, thrombotic events and mortality between the two groups. Analysis on blood loss, anaphylactic reactions and renal function was not performed in this study due to a lack of sufficient information.CONCLUSION: Aprotinin can reduce the intraoperative requirement for blood products in OLT, and has no significant effect on the incidence of laparotomy for bleeding, thrombotic events and mortality.

Effects of Aprotinin on Serum Interleukin-2 and Soluble Interleukin-2 Receptor during Cardiopulmonary Bypass

Interleukin-2 and its receptor are of importance in regulating immunity responses. The changes of interleukin-2 (IL-2) and soluble interleukin-2 receptor (IL-2R) during heart valve (s) replacement operation and effects of aprotinin on them were observed. Twenty patients undergoing heart valve (s) replacement were randomly divided into two groups: control group (n=10) and apro- tinin group (n=10). In aprotinin group, 1 000 000 KIU aprotinin was given by vein injection and then 2 000 000 KIU was given as a bolus in prime. Blood samples were collected before CPB, right after CPB and on the 1st, 3rd and 7th postoperative day (POD) for serum IL-2 and sIL-2R determination. Results showed that after CPB, IL-2 was reduced and slL-2R increased. Meanwhile, serum IL-2R was lower in aprotinin group than that of control. It is concluded that the immunity depression after CPB is associated with low level of IL-2 and high level of sIL-2R and aprotinin can ameliorate the situation.

Experimental Study on the Effects of Aprotinin on Myocardial Ischemia and Reperfusion

Direct effects of a high-dose aprotinin on the normally perfused hearts and the myocardial protection after ischemia and reperfusion were investigated in an isolated working rat heart model. In trial I, hearts had no ischemia and were perfused with either K-H solution or the K-H solution containing aprotinin (200KIU/ml) for 55 min. No statistically significant difference was observed in hemodynamics betweem the two groups. In trial Ⅱ, hearts were exposed to 150 minperiod of global ischemia at 15℃ with 4℃ multidose St. Thomas'Ⅱ solution (STS). The control group I received norma1 K-H solution; the group Ⅱ was treated with the solution with aprotinin added. The group, was similar to the group Ⅰ and received the STS enriched with aprotinin. On reperfusion, the recovery of hearts in group, was significantly better than those of the group Ⅰand Ⅱ, as reflected by better hemodynamics and myocardial oxygen consumption,lower level myocardial enzymes, higher myocardial ATP levels and milder myocardial ultrastructural injury. There was no difference between the group Ⅰand Ⅱ. These results suggest that the aprotinin at a dose of 200 KIU/ml has no harmful effects on normally perfused hearts and has a marked myocardial protective effect on the prolonged myocardial ischemia when used in cold crystalloid cardioplegia.

MECHANISM OF PRESERVING EFFECT OF APROTININ ON PLATELET FUNCTION DURING CARDIOPULMONARY BYPASS

The deficiency of platelet function is the main defect of hemostatic mechanism during cardiopulmonary bypass (CPB), which attributed to the postoperative bleeding complication to a great extent. The proteinase inhibitor aprotinin was reported to have preserving effect on platelet adhesion during CPB. In this clinical reserch we found that CPB caused plasma alpha 2-antiplasmin decreasing, indicating the fibrinolytic system activation. Meanwhile, the ristocetin-induced aggregation declined to 39.6% and platelet GPIb decreased to 50% of preoperative value. However, by treatment with aprotinin, the plasma alpha 2-antiplasmin during CPB did not change, platelet aggregation was improved and platelet GPIb was preserved, and consequently resulted in a 46% lower blood loss postoperatively. These results confirmed that aprotinin could inhibit the fibrinolysis during CPB, and thus relieve the platelet damage and improve the postoperative hemostatic mechanism.

THE IMPAIRMENT OF PLATELET FUNCTION IN FIBRINOLYSIS AND PRESERVING EFFECT OF APROTININ

Platelet adhesion depends on the platelet membrane glycoprotein Ib (GPIb) and plasma von Willebrand Factor (vWF), which can be reflected by ristocetin-induced aggregation. Here we report damage effect of fibrinolysis and preserving effect of aprotinin on platelet function. Addition of 40 U/ml urokinase and 0.3 U/ml plasmin to PRP or washed platelets made the ristocetin-induced aggregation decline to 31.6% and 38.5% of control value respectively. The extent of declining was positively correlated with the concentration of urokinase and plasmin. Meanwhile, the platelet GPIb decreased to 76.4% of control value. The results showed that the fibrinolysis impaired the platelet function and this effect may be associated with the hydrolysis of GPIb. Further research found that by adding the same dose of urokinase or plasmin to aprotinin-pretreated PRP or washed platelets, the aggregation did not change statistically and decrement of GPIb is much less marked. We concluded that the aprotinin could relieve the platelet dsfunction effectively by its inhibitory effect on fibrinolytic activity.

A clinical study of aprotinin blood anesthesia used in the surgery of liver cancer

Objective: To investigate the role of aprotinin blood anesthesia used in hepatotomy. Methods: Patients with liver cancer undergoing hepatotomy were divided into two groups. In experimental group (40 patients) a loading dose with 1112 EPU aprotinin and maintained by 278 EPU/h was used until 2 h after operation. The control group (42 patients) was treated with 0.9% normal saline. The venous blood was withdrew for blood routine, thrombelastography and coagulable test at the time of preinduced, 1 h, 2 h and 4 h following the operation beginning, 6 h and 12 h after operation. The change of TEG and coagulable profile were monitored during the whole surgery. The volume of blood transfusion and hemorrhage between two groups were compared. Results: After the usage of aprotinin, the preoperative hypercoagulability of the experimental group was remitted and the coagulative state was kept relatively stable during the operation. However, hypercoagulability of the control group aggravated following the operation beginning and some of them switched to hypocoagulability. The volumes and rates of hemorrhage and transfusion were smaller in the experimental group than in the control group. Conclusion: Aprotinin can stabilize the coagulable state, reduce the volumes and rates of hemorrhage and transfusion, and is worth using in the surgery of operations of liver cancer.

The influence of aprotinin on the mRNA expression of P-selectin and ICAM-1 in the in situ lung tissue after ischemic cold storage and reperfusion

To explore the influence of aprotinin on the mRNA expression of P-selectin and ICAM-1 in lung tissue after ischemic reperfusion.Methods Thirty New Zealand white rabbits were randomly divided into 3 groups:control group,LPD group and aprotinin group.In situ rabbit lung preservation model was established.In control group,the left lower lung lobe was stored at 10℃ in a specially made lung preservation container for 2 hours and reperfused for another 2 hours.In LPD group and aprotinin group,the left lower lobe was perfused with LPD solution or aprotinin containing LPD solution,respectively,after left lung hilus was clamped.The other procedures were the same as those in control group.The lung tissue was harvested at different time intervals including preclamping lung hilus,2 hours after clamping and 2 hours after reperfusion.The mRNA expression of ICAM-1 and P-selectin in the lung tissue was detected with RT-PCR technique.Results The contents of mRNA of P-selectin at 2 hours after reperfusion in control group and LPD group were significant higher than pre-ischemia and 2 hours after champing the left lung hilus.There was no such significant difference in aprotinin group.The mRNA expression of P-selectin in aprotinin group at 2 hours after reperfusion was significantly lower than that in control group and LPD group.The ICAM-1 mRNA expression at 2 hours after Ischemia and 2 hours after reperfusion in control group and LPD group was significantly higher than the pre-ischemia and its was significantly higher than that in aprotinin group.Conclusion Aprotinin can inhibit the upregulation of the mRNA expression of P selectin and ICAM-1 after ischemia reperfusion in the lung tissue,so the addition of aprotinin in LPD solution may reduce the ischemia reperfusion injury in lung tissue.5 refs,1 tab.

The evaluation of aprotinin contained preservation solution with a new animal model

ObjectiveTo explore the protective effect of aprotinin contained LPD (low potassium dextran) solution via an in situ rabbit lung preservation model.MethodsThirty New Zealand rabbits were divided randomly into 3 groups,10 in each group.In group A (control group),the left lung hilus was clamped without solution perfusion;In group B (LPD group) and group C (aprotinin group),the lungs were perfused with LPD solution and aprotinin contained LPD,respectively.The lungs in all groups were stored at 10 centigrade in a specially made lung preservation container for 2 hours and then unclamped the lung hilus to reperfuse the lung for 2 hours.Pulmonary venous blood samples were collected at pre-clamping of lung hilus,5 minutes and 120 minutes after reperfusion for analysis of blood gas.Biopsy of lung tissue was excised for morphological examination at pre-unclamping of lung hilus and 2 hours after reperfusion.Examination of bronchoalveolar lavage fluid was taken for the evaluation of inflammation status.ResultsPulmonary venous partial pressure of oxygen (PvPO2) in the 3 groups at 5 minutes and 120 minutes after reperfusion were significantly higher than those before clamping of lung hilus,respectively.PvPO2 in group A and group B at 120 minutes after reperfusion were significantly higher than those at 5 minutes after reperfusion.There was no significant difference of PvPO2 in group C between 5 minutes and 120 minutes after reperfusion.PvPO2 in group C at 5 minutes and 120 minutes after reperfusion were significantly higher than those in group A and group B.The morphological lesion was more severe in group A and B than that in group C.The PMN percentage in bronchoalveolar lavage fluid in group A and B was significantly higher than that in group C.ConclusionsThe protective effect of aprotinin is obvious for lung protection in animal model.Aprotinin contained lung preservation solution is superior to LPD for lung protection.

Effect of Enzymes in Buccal Mucous Membrane on Buccal Absorption of Insulin

To evaluate the effect of proteolytic enzymes on the absorption of insulin in the buccal mucosa, the trichloroacetic acid (TCA) method was used to estimate the degradation of insulin under different conditions in the buccal mucosal homogenates. In vivo experiments estimating the enhancement of hypoglycaemic effect by enzyme inhibitors were also conducted. The results showed that proteolytic enzymes in the buccal mucosa were less active than in the intestine. Bacitracin, aprotinin and sodium deoxycholate could inhibit the degradation of insulin in the buccal mucosal homogenates. The degradation of insulin in buccal mucosal homogenates of normal hamsters was smaller than that of diabetic hamsters. In vivo experiments of hypoglycaemia supported the in vitro results. When given buccally, bacitracin, aprotinin and sodium deoxycholate could increase the relative pharmacological bioavailability of insulin. When co-administered with aprotinin(0.1%), bacitracin(0.5%) and sodium deoxycholate(5%), the relative pharmacological bioavailabilities of insulin were 4.84%, 6.60% and 14.95% respectively. The in vitro and in vivo results suggest that proteolytic enzymes are present in the buccal mucosa, which limit absorption of insulin. Co-administration with some enzyme inhibitors can improve the bioavailability of insulin via buccal delivery and sodium deoxycholte is more efficient than some enzyme inhibitors used for improving buccal absorption.

丝氨酸蛋白酶抑制剂可抑制昆虫细胞表达的蛋白质降解(英文)

干细胞因子是一种多功能细胞因子,能在多级造血水平与其他细胞因子协同作用促进造血干/祖细胞及各种血细胞的存活、增殖和分化。重组人二连体干细胞因子具有比干细胞因子单体更高的比活,可以避免副反应。重组人二连体干细胞因子在Sf9细胞和大肠杆菌中表达时,发现有特异性降解。片段缺 失实验证实切割位点位于重组人二连体干细胞因子的145位到165位氨基酸之间。丝氨酸蛋白酶抑制剂aprotinin和PMSF能抑制这种特异性降解。试验了不同浓度的aprotinin和PMSF对Sf9细胞存活和重组人二连体干细胞因子产量的影响,显示当aprotinin的浓度为 1.0μg/mL时,重组人二连体干细胞因子的产量是没有加aprotinin时产量的2倍,而且aprotinin可以完全抑制这种特异性降解。

新生隐球菌的胞外蛋白水解酶及丝氨酸蛋白酶活性检测

目的:测定不同孵育条件、来源和血清型的新生隐球菌分泌的胞外蛋白水解酶及丝氨酸蛋白酶的活性。方法:30℃和37℃下用含有偶氮白蛋白的琼脂培养板培养不同来源及血清型的36株新生隐球菌菌株,测量其产生的廓清晕环的大小,计算CH值[菌落半径/(菌落半径+廓清晕环半径)]来比较其胞外蛋白水解酶及丝氨酸蛋白酶的活性强弱;用丝氨酸蛋白酶的特异性抑制剂观察两种酶活性改变;酶谱法观察菌株浓缩上清液中胞外蛋白水解酶活性和相对分子质量。结果:36株菌株在30℃和37℃培养条件下它们的平均CH值分别为0.558±0.170和0.575±0.169,两者间无显著差异;血清A(n=13)、B(n=13)、D/AD(n=6)型菌株各自的平均CH值分别为0.564±0.144、0.515±0.078和0.482±0.072,各组间无显著差异;临床分离株(n=23)、环境分离株(n=9)和荚膜缺陷株(n=4)各自的平均CH值分别为0.570±0.177、0.513±0.069和0.942±0.075(P<0.05)。对照组(不含抑肽酶)和抑肽酶不同浓度组(1.2、1.6 mU)的平均CH值分别为0.459±0.188,0.975±0.287、0.733±0.252(P<0.01)。菌株浓缩液中含有复杂的胞外蛋白酶类成分。结论:新生隐球菌胞外蛋白水解酶及丝氨酸蛋白酶的活性在30℃、37℃下无显著差异;在临床分离株中比环境分离株和荚膜缺陷株强;在不同血清型菌株间无显著差别;可被丝氨酸蛋白酶抑制剂抑制。

增强组织工程支架:纤维蛋白胶稳定性的初步研究

目的:研究抑肽酶和氨甲环酸对于减缓纤维蛋白胶支架降解速度的作用。方法:将抑肽酶和氨甲环酸加入纤维蛋白胶中,构建细胞标准支架复合物和细胞增强支架复合物,体外培养,观察软骨细胞在支架中的繁殖情况、形态学、超微结构检测以及载体降解情况。结果:两组复合物体外培养一二周后,其质量有显著性差异;两组复合物的细胞生长曲线无明显差别;软骨细胞在纤维蛋白胶支架中立体培养可分泌胞外基质。结论:在纤维蛋白胶中加入抑肽酶和氨甲环酸,可显著的减缓纤维蛋白胶的降解速度,并且对软骨细胞的繁殖、表型的维持、基质的分泌无不良影响。

口服抑肽酶对大鼠急性肝损伤的保护作用

目的:探讨口服抑肽酶对实验大鼠肝损伤的保护作用。方法:Wistar大鼠60只,采用四氯化碳(CCl4)法建立急性肝损伤实验动物模型,随机分为对照组、模型组及抑肽酶大、中、小剂量组和阳性对照组(甘利欣组),观察抑肽酶对实验鼠血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、一氧化氮(NO)、还原型谷胱甘肽(GSH)含量、肝重量系数以及肝组织病理改变的影响。结果:模型组与正常对照组比较,血清ALT及AST均显著增高(P<0.001)、NO明显增高(P<0.01)、肝重量系数明显增加(P<0.001)、GSH明显降低(P<0.01)。抑肽酶小、中、大剂量组与模型组比较:ALT及AST值均显著降低(P<0.001),GSH值均明显增高(P<0.01),与甘利欣组相近;NO值均明显降低(P<0.05),且低于甘利欣组;肝重量系数均明显降低(P<0.01,P<0.05,P<0.001)。鼠肝组织的病理性损伤改变程度,抑肽酶大、中、小剂量组及模型组依次加重。结论:口服抑肽酶对大鼠急性肝损伤具有明显的保护作用。

原位肝脏移植术中血液保护的临床研究

目的 研究原位肝移植手术中的血液保护。方法 35例行原位肝移植手术的病人 ,随机为分A、B两组。A组 2 0例 ,术中采用洗涤式自体血回收和大剂量抑肽酶的血液保护措施 ;B组 15例 ,不使用上述血液保护方法。A、B两组均加强血液动力学、体温、凝血功能监测包括活化部分凝血酶时间 (APTT)、凝血酶时间 (TT)、凝血酶原时间 (PT)、纤维蛋白原定量 (FIB)、激活全血凝血时间(ACT)、凝血弹性图 (TEG) (取R值作为观察指标 )。并统计两组术中失血量和输血量、A组回收的血量和回输的洗涤红细胞量。两组病人均在无肝期采用静脉 静脉转流。术中保持正常体温并于新肝期中和肝素。结果 A组平均出血量 (315 2± 16 35 )ml,B组 (4 2 6 5± 15 45 )ml(P <0 0 5 )。A组平均每例回收血量 (2 6 6 2± 1135 )ml,回输洗涤红细胞 (1310± 710 )ml。A组平均输血量 (2 410±95 0 )ml,B组 (2 886± 10 10 )ml(P <0 0 5 )。在凝血功能各项中 ,转流 30、6 0分钟、新肝期 15分钟及术毕时 ,A组各指标均优于B组 (P <0 0 5 ,P <0 0 1)。结论 原位肝移植术中应用相应的血液保护方法能节省用血 ,加强凝血功能的监测并予针对性处理 。

抑肽酶佐治脑出血的临床研究

目的 观察抑肽酶辅助治疗脑出血脑水肿的疗效及安全性。方法 收集脑出血患者 41例 ,分为 2组 ,抑肽酶治疗组 2 1例 ;对照组 2 0例。治疗前后进行神经功能评分 ,CT下测量水肿体积。结果 (1 )入院后 1、2、3周用欧洲脑卒中量表进行神经功能评分 ,抑肽酶治疗组分值高于对照组 (P <0 .0 5)。 (2 )水肿产生量治疗组少于对照组 (P <0 .0 5)。结论 脑出血急性期加用抑肽酶可以抑制脑水肿的形成 ,提高脑出血患者神经功能评分 。

不同剂型药用抑肽酶纯度的胶束电动毛细管色谱测定

以十六烷基三甲基溴化铵 (CTAB)为阳离子表面活性剂 ,用胶束电动毛细管色谱 (MECC)分别对抑肽酶粉针剂和抑肽酶注射液进行纯度测定。实验中选择了最佳缓冲液 (含 4mmol/LCTAB的 80mmol/LNa2 HPO4 H3 PO4,pH 7 0 0 ) ,考察了进样量与样品中高浓度盐对分离的影响。并对毛细管区带电泳、MECC和高效液相色谱的分离效果加以比较 ,表明MECC的分离效果最佳。

氨甲环酸对心脏手术后出血量及并发症影响的对照研究

目的:观察氨甲环酸对心脏手术后出血量的影响。方法:根据临床停止使用抑肽酶的时间,将560例行心脏手术患者分为抑肽酶组(n=303)和氨甲环酸组(n=257),对两组术后失血量、二次开胸止血率、输血量、急性肾功能不全及低心排综合征发生率等进行比较。结果:术后24 h出血量氨甲环酸组为(578.5±386.0)ml,抑肽酶组为(629.3±366.7)ml,两组差异无统计学意义(P=0.111)。二次开胸止血率氨甲环酸组为3.9%,抑肽酶组为4.3%,两组差异无统计学意义(P=0.730)。术后红细胞需求量氨甲环酸组为(2.39±6.20)U,抑肽酶组为(2.08±4.92)U;血浆需求量氨甲环酸组为(1.58±4.85)100 ml,抑肽酶组为(1.67±4.88)100 ml;血小板需求量氨甲环酸组为(1.19±5.12)10 U,抑肽酶组为(1.51±5.29)10 U,两组差异均无统计学意义(分别P=0.149、0.355、0.797)。结论:心脏手术中应用氨甲环酸可有效减少术后出血量及血制品需求量,其效果与抑肽酶相似。