少弱精子症和正常生育男性精液中尿激酶及其受体含量的研究

目的:通过研究精子正常和异常男性精浆和精子中尿激酶及受体含量差异,以了解尿激酶及受体与男性生育力的关系。方法:采用双抗体夹心ELISA法测定22例正常生育男性和44例少弱精子症男性精浆和精子中尿激酶及受体的含量。结果:①正常男性精浆尿激酶平均含量为(4 803.69±602.78)mU/L,与少弱精子症组[(4 061.35±736.23)mU/L]相比,差异有显著性(P<0.01)。正常生育男性精子尿激酶平均含量为(30.29±3.16)mU/106个精子,与少弱精子症组[(20.51±4.2)mU/106个精子],差异有显著性(P<0.01)。②正常生育男性精子尿激酶受体平均含量为(12.97±3.11)mU/106个精子相比,与少弱精子症组[(6.09±1.45)mU/106个精子]相比,差异有显著性(P<0.01)。③精子和精浆中尿激酶含量和精子活率和活力呈显著正相关。结论:尿激酶和男性生育力相关,少弱精子症和正常生育男性精液中尿激酶及其受体含量存在差异。     

Determination of Tissue Plasminogen Activator (t-PA) Antigen in Seminal Plasma by Modified Enzyme-Linked Immunosorbant Assay (ELISA)

Summary: The determination of tissue plasminogen activator (t-PA) antigen in semen by a new immunoenzymatic method was done in 152 men without genito urinary pathology. The seminal value is one hundred higher than the blood value with a log normal distribution. These levels are not influenced by freezing temperature, by the sequences freezing-thawing, by centrifugation. From split ejaculates a correlation has been established between t-PA antigen and zinc and not with fructose. These results indicate the major prostatic origin of the t-PA antigen.
Zusammenfassung: Bestimmung des Gewebe-Plasminogen-Aktivator-Antigens im Spermaplasma mittels modifizierter Elisa-Methode
Bei 152 Männern ohne kongenitale Erkrankung des Urogenitaltraktes wurde eine Bestimmung des Gewebe-Plasminogen-Aktivator-Antigens (t-PA Antigen) im Sperma mittels einer neuen enzymimmunologischen Methode durchgeführt. Die Werte im Sperma liegen hundertmal höher als diejenigen im Blut bei einer logarithmischen Normalverteilung. Die Werte werden weder durch Einfrieren noch durch wiederholtes Einfrieren und Auftauen im Wechsel, noch durch Zentrifugieren beeinflußt. Im Split-Ejakulat konnte eine Korrelation zwischen tPA-Antigen und Zink, nicht jedoch zwischen dem t-PA-Antigen und Fruktose festgestellt werden. Diese Ergebnisse zeigen, daß der größte Teil des t-PA Antigens prostatischen Ursprungs ist.     

Fibrinolytic parameters in spermatozoas and seminal plasma

Urokinase-type (u-PA) and tissue-type plasminogen activator antigen (t-PA) as well as plasminogen activator-inhibitor activity were determined in seminal plasma and lysates of the respective spermatozoas in 67 ejaculate of males in infertile marriage without genito urinary pathology. U-PA was determined by a competition RIA, t-PA by an ELISA and PAI by a spectrophotometric assay. 15 patients showed normozoospermia, 11 azoospermia and 41 oligoasthenoteratozoospermia (OAT-syndrome). In lysates of spermatozoas, significantly higher levels of both plasminogenactivators and PAI were found in patients with OAT syndrome as compared to those exhibiting normozoospermia. Whereas PAI was absent in the seminal plasma of normozoospermic ejaculate, patients with azoospermia (180 +/- 13 mU/ml.) and OAT-syndrome (60 +/- 5 mU/ml.) showed high PAI levels. The similarly high values of t-PA (190.8-227.8 ng./ml.) and u-PA (19.4-32 ng./ml.) in the same compartment confirm their predominantly prostatic origin and seem to have no influence on the quality of the ejaculate.     

尿激酶在雄性生殖中的作用

尿激酶型纤溶酶原激活因子(uPA)存在于许多动物的生殖系统,在雄性生殖系统精子和精浆中都可以检测到。关于uPA的作用机制尚未完全阐明,目前多数学者认为uPA是通过与uPA受体结合而发挥作用。在雄性生殖系统,uPA的生理作用是多方面的,它参与精子发生、成熟、运动、获能、顶体反应、受精以及精液液化等生殖生理过程。现就uPA在此方面的研究进展作一综述。     

Peritoneal fibrinolytic activity in peritonitis

BACKGROUND: Peritonitis may cause a reduction in abdominal fibrinolytic activity. The reduced local fibrinolysis seems to be an important process in the subsequent development of adhesion formation. The aim of the study was to evaluate peritoneal fibrinolytic capacity in inflamed and normal peritoneum. METHODS: Peritoneal biopsy specimens were taken at the beginning of operation from 15 patients with peritonitis and 10 patients who underwent elective operation. Levels of tissue-type plasminogen activator (tPA), urokinase-type plasminogen activator (uPA), plasminogen activator inhibitor (PAI) type 1 (PAI-1) and type 2 (PAI-2), and tPA/PAI complex in tissue extracts were determinated by commercially available enzyme-linked immunosorbent assay kits. RESULTS: tPA was significantly reduced in peritonitis compared with normal peritoneum (P <0.001), whereas it was found that the levels of PAI-1, PAI-2, uPA, and tPA/PAI complex in peritonitis were significantly higher than those in normal controls. CONCLUSIONS: Plasminogen activator activity was significantly reduced in peritoneal biopsy samples from patients with peritonitis compared with those from patients without peritonitis.     

Breed and seasonal variation of plasminogen activator activity and plasminogen activator inhibition in spermatozoa and seminal plasma of the ram in correlation with testosterone in the blood

Summary. Plasminogen activator activity (PAA) and plasminogen activator inhibition (PAI), against t-PA (t-PAI) or u-PA (u-PAI), in spermatozoa and seminal plasma as well as testosterone in the blood of Friesland, Chios, and Karagouniki rams all showed a seasonal variation with the highest values during the corresponding breeding season of the ewes (Autumn-Winter). The seasonal variation of PAA and PAI in spermatozoa or seminal plasma as well as blood testosterone was different among the three breeds studied. Increased spermatozoal PAA was observed in November and May in Friesland rams, in October and November in Chios rams, and in October in Karagouniki rams. Spermatozoal t-PAI was increased in December and June in Friesland rams, in November and December in Chios rams, and in November in Karagouniki rams. Spermatozoal u-PAI was increased in December in Friesland rams, in October and December in Chios rams, and in November and December in Karagouniki rams. Plasminogen activator activity and PAI in seminal plasma also showed similar seasonal variations. Plasminogen activator activity and PAI in spermatozoa and seminal plasma showed a positive correlation with blood testosterone. The results of the present study support our previous findings on the possible role of spermatozoal PAA and PAI in the fertilizing ability of spermatozoa.     

Enhanced fibrinolytic activity during the course of hemodialysis.

In order to clarify the effect of hemodialysis (HD) on the fibrinolytic system, fibrinolytic activity was evaluated in 27 patients undergoing regular hemodialysis treatment (RDT) using new parameters including plasma alpha 2-plasmin inhibitor (alpha 2 PI), alpha 2-plasmin inhibitor-plasmin complex (alpha 2 PIC), cross-linked fibrin degradation products (XL-FDP), tissue plasminogen activator (t-PA) activity, t-PA antigen and plasminogen activator inhibitor-1 (PAI-1) antigen. Predialysis baseline levels of plasminogen and alpha 2PI activity in RDT patients were significantly lower and those of alpha 2PIC were significantly higher than normal control values. During a single HD session, alpha 2PIC exhibited a continuous, significant increase reaching about 180% of initial values by the end of HD. alpha 2PI activity was significantly decreased at the end of the HD, though there were no significant changes in plasminogen activity during HD. Predialysis baseline levels of XL-FDP in RDT patients were significantly higher than normal control values. No significant changes in XL-FDP were observed during HD. Both t-PA activity and t-PA antigen significantly increased during HD, and PAI-1 antigen significantly decreased during HD. Von Willebrand factor (vWF) antigen in plasma, which is regarded as reflecting a release reaction by vascular endothelial cells to certain stimuli, also significantly increased during HD. However, neither vWF antigen nor t-PA antigen was increased by heparin administration alone. The changes in alpha 2PI and alpha 2PIC levels suggest that fibrinolytic activity is slightly higher in RDT patients and is even higher during HD.(ABSTRACT TRUNCATED AT 250 WORDS)     

The relationship between cardiovascular complications of estrogen therapy and fibrinolysis in patients with prostatic cancer.

To determine the relationship between cardiovascular complications of estrogen therapy and fibrinolysis, fibrinolysis parameters plasminogen, urokinase-type plasminogen activator (u-PA), tissue-type plasminogen activator (t-PA), and plasminogen activator inhibitor-1 (PAI-1), were assessed in 12 prostatic cancer patients before and 6 weeks after the onset of estrogen therapy. The levels of plasminogen, u-PA, and PAI-1 in the patients treated with the estrogen therapy were significantly higher than those in the patients before the therapy. The t-PA level in the patients during the therapy was significantly lower than that before the treatment. Cardiovascular complications were found in two patients (16.7%) during estrogen therapy. In the two patients, marked elevation of PAI-1 and decreased level of t-PA were observed during the therapy. These results indicate that cardiovascular complications of estrogen therapy in patients with prostatic cancer may be related to hypofibrinolysis resulting from changes of PAI-1 and t-PA.     

多囊卵巢综合征患者血纤溶能力的研究

目的了解多囊卵巢综合征(PCOS)患者的血纤溶能力及服用二甲双胍后血纤溶能力的变化。方法(1)对照组20例、PCOS患者30例,分别取血检测组织纤溶酶原激活物(t-PA)、纤溶酶原激活抑制物(PAI-1)的活性;(2)PCOS高胰岛素血症者10例,分别给予二甲双胍750~1 500 mg/d治疗,比较治疗前、后t-PA、PAI-1活性。结果PCOS组的血胰岛素(INS)、游离睾酮(F-T)t、-PA、PAI-1分别为(24.42±12.30)mU/L(、2.70±1.50)ng/L、(0.17±0.06)KU/L(、0.88±0.05)AU/ml,对照组分别为(10.04±6.12)mU/L(、1.70±1.00)ng/L(、0.28±0.04)KU/L、(0.70±0.09)AU/ml。10例PCOS患者经二甲双胍治疗后INS、F-Tt、-PA、PAI-1分别为(12.20±7.78)mU/L、(1.70±1.00)ng/L、(0.26±0.08)KU/L、(0.70±0.35)AU/ml。结论PCOS患者的血纤溶能力低于正常者,而服用二甲双胍后血纤溶能力明显改善。     

Production of Urokinase-Type Plasminogen Activator (u-PA) and Plasminogen Activator Inhibitor-1 (PAI-1) in Human Brain Tumours

Summary We investigated the role of plasminogen activators (PAs) and their inhibitor (plasminogen activator inhibitor-1, PAI-1) in human brain tumours. The amounts of urokinase-type plasminogen activator (u-PA), tissue-type plasminogen activator (t-PA), and plasminogen activator inhibitor-1 (PAI-1), and the activity of u-PA and t-PA were determined by enzyme-linked immunosorbent assay (ELISA), and u-PA and PAI-1 were immunolocalized using monoclonal antibodies in human brain tumours and normal brain tissues. The tissues were surgically removed from 64 patients; normal brain tissue (5 cases), low-grade glioma (4 cases), high-grade glioma (17 cases), metastatic tumour (9 cases), meningioma (benign 12 cases, malignant 6 cases), acoustic schwannoma (11 cases). u-PA activity and u-PA and PAI-1 antigen levels were significantly elevated in malignant brain tumours (malignant meningiomas, high-grade gliomas, and metastatic tumours) and acoustic schwannomas but very low in benign meningiomas, low-grade gliomas and normal brain. There was no difference in t-PA antigen levels among normal and malignant tissues, however levels of t-PA activity were markedly decreased in metastastic tumours. All malignant brain tumour tissues showed positive immunostaining for u-PA and PAI-1, however, some tumour cells showed negative intensity while others showed strong intensity for these antibodies. This contrasts to the homogeneous staining pattern found in acoustic schwannoma. These findings indicate that malignancy in human brain tumours is associated with elevated levels of u-PA and PAI-1 and that an imbalance between these proteins in a micro-enviroment contributes (ascribes) to tumour cell invasion.     

Increased plasminogen activator activity and plasminogen activator inhibition in spermatozoa and seminal plasma of the ram after serum gonadotrophin (PMSG) administration. Correlation with the increased level of testosterone in the blood

After a single injection of serum gonadotrophin (PMSG) at the dose of 15 IU/kg, i.m. into rams testosterone in the plasma of blood showed a significant rise between 4th and 7th day post-injection. At the same time (4th-7th day) the plasminogen activator activity (PAA) in seminal plasma was found to be increased, but the plasminogen activator inhibition (PAI) expressed against t-PA (anti-t-PA) showed an increase between 32nd and 46th day. In spermatozoa a marked increase of PAA was revealed between 32nd and 46th day post-injection, while an increase of PAI (anti-t-PA) was exhibited on the 74th day. Plasmin inhibition (PI) in seminal plasma and spermatozoa showed no change compared to controls. A positive correlation has been found between increased concentrations of testosterone and PAA or PAI (anti-t-PA) in spermatozoa and seminal plasma. The induced increase of PAA in spermatozoa under the effect of testosterone might be of physiological importance, since PAA is localized to sperm membranes and might participate in the whole process of fertilization.     

Comparison of hepatic coagulant, fibrinolytic, and anticoagulant functions between Banna Minipig Inbred line and humans

BACKGROUND: As an ideal candidate for xenotransplantation, the compatibility of physiological porcine organs with those of humans is an essential premise. In this study, we analyzed hepatic coagulant, fibrinolytic, and anticoagulant functions between Banna Minipig Inbreds (BMIs) and humans to evaluate such hepatic compatibility. METHODS: BMI factors II, V, VII, X, and XII were added to the corresponding factor-deficient human plasma to determine prothrombin times (PT) and activated partial thromboplastin times (APTT). Human tissue plasminogen activator (t-PA) was added to both BMI and human plasma to determine plasmin activity. The antithrombin-III (AT-III) activity of plasma was analyzed with the STA-Stago autoanalyzer using an AT-III assay kit. RESULTS: Both PT and APTT were reduced but within normal parameters when BMI factors II, V, VII, X, and XII were added to the corresponding factor-deficient human plasma. The activities of BMI coagulation factors II, V, VII, X, and XII were 3.2, 3.7, 4.7, 2.9, and 4.5 times those of humans, respectively. The activity of plasmin was significantly higher in BMI plasma than in humans when human t-PA was added to both. The normal range of human AT-III activity was 90-108% while BMI AT-III was 124.50 +/- 2.38%. CONCLUSIONS: The activities of coagulation factors and AT-III were higher in BMIs than in humans. BMI coagulation factors XII, VII, and X trigger human intrinsic, extrinsic, and common pathways, respectively, which functioned normally. In addition, BMI plasminogen could be activated by human t-PA.     

7/8肾切除大鼠凝血纤溶系统的变化及干预治疗的影响

目的：探讨肾小球硬化及小管间质纤维化过程中肾组织纤溶酶原激活物（PA）和PA抑制物（PAI）－1蛋白表达及干预治疗的影响。方法：7／8肾切除肾功能衰竭大鼠为实验动物模型,随机分为未治疗组和治疗组,观察12周后各组大鼠血尿各项生化指标、尿PA活性及残肾组织常规病理和用免疫组织化学染色定性定量评价残肾组织型PA（tPA)、尿激酶型（uPA)、PAI－1蛋白表达。结果：未治疗组大鼠肾功能进行性丧失,尿PA活性下降,肾组织PAI－1表达增高,而tPA、uPA表达下降,残肾组织出现肾小球硬化和间质纤维化。治疗组大鼠残肾组织tPA、uPA蛋白表达增加,PAI－1表达下降。尿PA活性增加,肾功能改善。结论：水蛭治疗组、苯那普利治疗组及联合治疗组都可以通过改善7／8肾切除大鼠PA／PAI－1系统的紊乱而延缓肾小球硬化和间质纤维化病变的进展。     

Plasma fibrinolytic activity in patients undergoing major abdominal surgery

The fibrinolytic system was studied in 30 patients undergoing elective cholecystectomy and in 30 with more advanced elective abdominal surgery. Blood was sampled before, during and after operation for determination in plasma of the tissue plasminogen activator (t-PA), the recently described fast t-PA inhibitor, and plasmin alpha 2-antiplasmin complex (PAP). In addition the t-PA activity during venous occlusion was determined preoperatively. Most of the patients showed raised t-PA levels during surgery, but the interindividual variation was wide and was not correlated to fibrinolytic capacity measured preoperatively as enhancement of t-PA activity during venous occlusion. The levels of the t-PA inhibitor rose during and immediately after surgery, and were higher in patients without increased t-PA activity during surgery. The patients with more advanced disease had higher levels of the inhibitor than the cholecystectomy patients. The data suggest that the t-PA inhibitor may influence the fibrinolytic response to surgical trauma and may explain the previously reported shutdown in fibrinolysis in the early postoperative period. PAP, used as reflecting the overall fibrinolytic activity, was increased in plasma after the first postoperative days.     

Effect of dextran on plasma tissue plasminogen activator (t-PA) and plasminogen activator inhibitor-1(PAI-1) during surgery

Dextran is known to increase the plasminogen activation rate in vitro and to decrease the α₂-antiplasmin activity.
We decided to explore the effect of dextran on plasma tissue plasminogen activator (t-PA) and plasminogen activator inhibitor-1(PAI-1) during surgical trauma.
Thirty-one patients undergoing elective surgery were given 500 ml of 6% dextran 70. Another nine patients serving as controls were given 500 ml of a glucose-electrolyte solution. The activities of t-PA and PAI-1during surgery were determined, as was the concentration of t-PA antigen.
PAI-1activity was decreased by 19% after infusion of 250 ml of dextran. After 500 ml, the activity was reduced by 22% (both P <0.05). The activity of t-PA was increased by 43% and 29% (both P <0.05) and the antigenic amount of t-PA was increased by 18% and 15% (both P <0.05) after infusion of 250 ml and 500 ml of dextran, respectively. No changes in these variables were observed in the control patients.
It is concluded that infusion of dextran promotes fibrinolysis by enhancing plasminogen activation in patients subjected to trauma. Since elevated levels of PAI-1prior to surgery are known to predispose to deep vein thrombosis, which may form already during the operation, the effect of dextran on PAI-1described here may explain its clot preventing properties.     

Enhanced fibrinolysis caused by tissue plasminogen activator release in hemodialysis

The effect of hemodialysis on components of the fibrinolytic system was measured in 22 patients. Plasma levels of t-PA antigen, t-PA activity, u-PA antigen and plasminogen activator inhibitor were determined by means of immunological and functional assays. During hemodialysis, 11 patients exhibited an increase in t-PA antigen within the first hour to about three times the starting values (P less than 0.05), followed by a decrease to about double of the initial values until the end of the treatment. Eleven patients showed a continuous increase up to 200% of the starting values until the end of hemodialysis. u-PA levels did not change significantly during the time of investigation (P greater than 0.5).     

Decreased fibrinolytic capacity in coronary patients by increased plasminogen activator inhibitor activity]

The fibrinolytic capacity of the blood mainly depends on the amount of tissue-plasminogen activator (t-PA) antigen and plasminogen activator inhibitor (PAI). In this study the fibrinolytic response to a venous occlusion test (VOT) was measured in 109 patients with angiographically documented coronary artery disease (CAD) and in 20 healthy volunteers at comparable age (controls). CAD-patients had higher plasma plasminogen activator inhibitor capacity before (24.4 +/- 11.0 vs. 15.4 +/- 5.2 arbitrary units [AU/ml]; p less than 0.0002) and after VOT (19.6 +/- 13.2 vs. 10.9 +/- 5.3 AU/ml; p less than 0.0001) compared with controls. Furthermore they showed significant lower plasma t-PA activity after VOT (3.0 +/- 6.8 vs. 6.6 +/- 10.6 AU/ml; p less than 0.0001). However there were no difference between both groups in plasma t-PA antigen levels after VOT (17.3 +/- 12.1 vs. 18.7 +/- 14.4 ng/ml). In 10% of patients the decrease in fibrinolytic activity resulted from a lower t-PA release ("lower" was defined as mean minus one standard deviation of the control group). 40% showed elevated plasma PAI capacity before VOT ("elevated" was defined as mean plus two standard deviations of the control group). Both caused significantly reduced post occlusion plasma t-PA activity and prolonged Euglobulin clot lysis time (p less than 0.003). A positive correlation was found between PAI capacity and serum triglyceride levels. Reduced fibrinolytic activity in 109 patients with coronary heart disease based either on a decrease in t-PA antigen release or a increased in PAI capacity in comparison with healthy controls. The mechanism of these findings is not yet well-known.(ABSTRACT TRUNCATED AT 250 WORDS)     

Increased fibrinolysis in orthotopic but not in heterotopic liver transplantation: the role of the anhepatic phase

The major cause of increased tissue-type plasminogen activator (t-PA) activity during orthotopic liver transplantation (OLT) is still unclear. Both lack of hepatic clearance of t-PA in the anhepatic period and/or increased endothelial release from the graft upon reperfusion have been suggested. Heterotopic liver transplantation (HLT) avoids resection of the host liver and is therefore a useful model to differentiate these two possibilities. The fibrinolytic system was evaluated in ten patients with OLT and in 18 patients with HLT. A marked increment in t-PA activity was observed during the anhepatic period of OLT, which rapidly normalized after reperfusion. In contrast t-PA activity levels remained normal in HLT. As a reflection of the increased t-PA activity fibrin degradation products were markedly elevated during OLT and plasminogen and α₂-antiplasmin decreased simultaneously during the anhepatic period. In conclusion, the lack of hepatic clearance during the anhepatic period is the most important factor in the evolution of increased t-PA activity during OLT.     

Effects of seprafilm on peritoneal fibrinolytic system

BACKGROUND: There is a high incidence of adhesions after ventral hernia repair with polypropylene mesh. The purpose of the present study was to evaluate the efficacy of Seprafilm in the prevention of adhesion formation and effect on peritoneal fibrinolytic activity. METHODS: An incisional hernia model was created in rats. In the experimental group Seprafilm was placed between polypropylene mesh and abdominal organs. On the 14th day adhesions were evaluated and tissue plasminogen activator (tPA), urokinase plasminogen activator (uPA), plasminogen activator inhibitor (PAI) type 1 and 2 were measured in peritoneal biopsy specimens. Results: Adhesions were significantly reduced in the Seprafilm group (P = 0.002). Nevertheless, there were no difference between the two groups in levels of tPA, PAI-1 and PAI-2. However, the levels of uPA were significantly decreased in the Seprafilm group. CONCLUSIONS: The adhesion preventive effect of Seprafilm is not directly related in peritoneal fibrinolytic activity. Instead, the physical properties (barrier, hydroflotation and sliconizing effect) of the membrane are primarily responsible for adhesion prevention.     

不同胃癌细胞系尿激酶型纤溶酶原激活物表达与腹膜转移潜能相关的体外研究

目的比较不同胃癌细胞系尿激酶型纤溶酶原激活物(uPA)表达及其活性与腹膜转移潜能的关系。方法采用ELISA和Western印迹方法,比较不同胃癌细胞系uPA表达的差异,并测定其活性。在24孔培养板或Boyden小室中与生长良好的间皮细胞共同培养不同时间,在显微镜下直接计数与间皮细胞黏附的胃癌细胞,而用MTT法评估胃癌细胞在间皮细胞间的迁移和侵袭情况。结果4株胃癌细胞(AGS、SGC7901、MKN45和MKN28)的uPA表达以SGC7901最高,uPA活性以MKN45最高,AGS两者皆最低。MKN45的黏附能力明显强于MKN28(P<0.05)、SGC7901(P<0.05)和AGS(P<0.01),但其迁移和侵袭能力与SGC7901和MKN28相比差异无统计学意义;而AGS在3方面均明显弱于其他3株细胞。结论4株胃癌细胞uPA表达量及其活性差异较大,并且与其腹膜转移潜能呈正相关。