Estudo imuno-histoquímico e in vitro de proteínas do sistema ativador de plasminogênio em carcinomas de células escamosas de língua oral / Immunohistochemical and in vitro study of plasminogen activator system proteins in oral tongue squamous cell carcinomas

O carcinoma de células escamosas (CCE) é a neoplasia maligna mais frequente da cavidade oral e apresenta prognóstico desfavorável. Assim sendo, pesquisas têm buscado esclarecer o papel de biomarcadores no comportamento biológico do CCE oral. Nesta perspectiva, destacam-se o ativador de plasminogênio tipo uroquinase (uPA) e seu receptor (uPAR), além do inibidor do ativador de plasminogênio-1 (PAI-1). O presente trabalho analisou, por meio de imuno-histoquímica, a expressão das proteínas uPA, uPAR e PAI-1 no CCE de língua oral (CCELO) e sua relação com parâmetros clinicopatológicos. Este experimento também avaliou os efeitos in vitro da proteína recombinante humana PAI-1 (rhPAI-1) na linhagem celular SCC25, derivada de CCELO. A imunoexpressão de uPA, uPAR e PAI-1 foi analisada em 60 casos de CCELO, de forma semiquantitativa, nas células neoplásicas do front de invasão tumoral. Visando a associação dos achados imuno-histoquímicos com variáveis clinicopatológicas e taxas de sobrevida, os casos foram classificados nas categorias baixa expressão (≤50% das células positivas) e alta expressão (>50% das células positivas). No experimento in vitro, foram analisados os seguintes grupos: G0 (controle; células cultivadas na ausência de rhPAI-1), G10 (células tratadas com rhPAI-1 a 10 nM) e G20 (células tratadas com rhPAI-1 a 20 nM). Diferenças entre estes grupos foram investigadas através dos ensaios: viabilidade celular (Alamar Blue), ciclo celular (marcação com iodeto de propídio, PI), apoptose/necrose (marcação com Anexina V e PI), atividade migratória (Wound healing) e invasão celular (Transwell). A análise imuno-histoquímica revelou alta expressão do uPA na maioria dos CCELOs, mas sem relações significativas com parâmetros clinicopatológicos. As expressões do uPAR e do PAI-1, em nível membranar, foram associadas a recidivas locais (p=0,019) e ao elevado tumor budding (p=0,046), respectivamente. A expressão membranar do PAI-1 também apresentou associação significativa com o alto escore de risco histopatológico (p=0,043). A análise estatística evidenciou ausência de associações significativas entre as variáveis imunohistoquímicas (uPA, uPAR e PAI-1) e indicadores de prognóstico do CCELO (sobrevida específica e sobrevida livre da doença). No estudo in vitro, decorridas 24 horas da administração da rhPAI-1, os grupos G10 e G20 exibiram maior viabilidade celular em comparação ao grupo controle (p=0,020), assim como aumento da progressão para a fase S do ciclo celular (p=0,024). No que concerne aos percentuais de células apoptóticas e necróticas, não foram encontradas diferenças significativas entre os grupos. Nos grupos celulares cultivados na presença da rhPAI1, também foi constatado aumento da atividade migratória (p=0,039) e do potencial de invasão (p=0,039), respectivamente, nos intervalos de 24 horas e 72 horas. Os achados deste estudo sugerem o envolvimento das proteínas uPA, uPAR e PAI-1 na patogênese do CCELO. Entretanto, a expressão destes biomarcadores pode não estar relacionada com a sobrevida dos pacientes. Os resultados in vitro demonstram que o PAI-1 exerce efeitos estimulatórios na proliferação, migração e invasão celular, podendo assim contribuir para a agressividade biológica do CCELO (AU).

Squamous cell carcinoma (SCC) is the most frequent malignant neoplasm of the oral cavity and has an unfavorable prognosis. Thus, studies have sought to clarify the role of biomarkers in the biological behavior of oral SCC. Within this context, urokinase-type plasminogen activator (uPA) and its receptor (uPAR), as well as plasminogen activator inhibitor 1 (PAI-1), are particularly interesting. The present study analyzed, by means of immunohistochemistry, the expressions of uPA, uPAR and PAI-1 in oral tongue SCC (OTSCC) and their relationship with clinicopathological parameters. This experiment also evaluated the in vitro effects of recombinant human PAI-1 (rhPAI-1) on the OTSCC-derived cell line SCC-25. The immunoexpression of uPA, uPAR and PAI-1 was analyzed semiquantitatively in neoplastic cells of the invasion front of 60 OTSCC cases. Aiming to determine the association between immunohistochemical findings, clinicopathological variables and survival rates, the cases were classified as low expression (≤50% of positive cells) and high expression (>50% of positive cells). The following groups were analyzed in the in vitro experiment: G0 (control; cells cultured in the absence of rhPAI-1), G10 (cells treated with 10 nM rhPAI-1), and G20 (cells treated with 20 nM rhPAI-1). Differences between these groups were investigated using the following assays: cell viability (Alamar Blue), cell cycle (staining with propidium iodide, PI), apoptosis/necrosis (staining with Annexin V and PI), migratory activity (Wound healing), and cell invasion (Transwell). Immunohistochemical analysis revealed high expression of uPA in most OTSCC cases, but there were no significant associations with clinicopathological parameters. The high membrane expression of uPAR and PAI-1 was associated with local recurrence (p=0.019) and high tumor budding (p=0.046), respectively. Membrane expression of PAI-1 also presented a significant association with high-risk cases (p=0,043). Statistical analysis demonstrated no significant associations between the immunohistochemical variables (uPA, uPAR and PAI-1) and prognostic indicators of OTSCC (disease-specific and disease-free survival). In the in vitro experiment, 24 hours after administration of rhPAI-1, G10 and G20 exhibited greater cell viability compared to the control group (p=0.02), as well as increased progression to the S phase of the cell cycle (p=0.024). There were no significant differences in the percentages of apoptotic or necrotic cells between groups. In the groups cultured in the presence of rhPAI-1, migratory activity (p=0.039) and invasion potential (p=0.039) were found to be increased after 24 and 72 hours, respectively. The findings of this study suggest the involvement of uPA, uPAR and PAI-1 in the pathogenesis of OTSCC. Nevertheless, the expression of these biomarkers may not be related to survival of patients. The in vitro results suggest that PAI-1 exerts stimulatory effects on cell proliferation, migration and invasion and may therefore contribute to the biological aggressiveness of OTSCC (AU).

Effects of Danhong Injection (丹红注射液) and its main components on anticoagulation and fibrinolysis in cultured vein endothelial cells / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effects of Danhong Injection (丹红注射液) and its main components, including daiclzein and hydroxysafflor yellow A (HSYA), on the anticoagulation, fibrinolysis, anti-apoptosis in hypoxia model of vein endothelial cells (VECs).</p><p><b>METHODS</b>VECs were prepared and were put in a hypoxia environment, which consisted of mixed gas of 95% N and 5% CO mixed gas, when reached confluent culture. Five groups used different treatments, including normal control group, hypoxia group, daiclzein group, HSYA group and Danhong Injection group. The VECs were identified by fluorescence double labeling methods. The morphology was observed by a phase contrast microscopy. The effects of Danhong Injection, daiclzein and HSYA on 6 keto prostaglandin F1α (6-keto-PGF1α) level was measured by the method of radioimmunoassay (RIA). Superoxide dismutase (SOD) activity was tested by water soluble tetrazolium salt. The content of malondialdehyde (MDA) was measured by thiobarbituric acid. The activities of tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI) were measured by the method of chromogenic substrate. The contents of endothelin (ET) and nitric oxide (NO) were detected by non-equilibrium RIA and enzymelinked immunosorbent assay. Cells apoptosis rate was determined by flow cytometry.</p><p><b>RESULTS</b>Compared with the normal control group, the floating cells number, PAI activity, ET and MDA contents, and cells apoptosis rate in the culture solution of hypoxia group were all significantly increased, whereas the 6-keto-PGF1α and NO contents, and t-PA and SOD activities were decreased significantly (P<0.01). Compared with the hypoxia group, Danhong Injection markedly increased the 6-keto-PGF1α content and SOD activity, regulated PAI and t-PA activities, ET and NO contents, and decreased MDA content and cells apoptosis rate (P<0.05 or P<0.01).</p><p><b>CONCLUSIONS</b>Danhong Injection and its main components played an important role in protecting primary VECs from hypoxic damage by regulating the secretion and vasomotor function of VECs. The function of Danhong Injection was most remarkable.</p>

Effect of thyroxine on plasminogen activator and inhibitor activity in rat.

Thyroid hormones influence mineral metabolism, distribution of water and electrolytes and are therefore of great importance in the maintenance of homeostasis under normal and diseased conditions such as renal failure. The present study was carried out to determine the effect of thyroxine on fibrinolytic parameters such as plasminogen activators (PA) in rat kidney, levels of PA and plasminogen activator inhibitor (PAI), glucose in plasma and serum lipid profile injected with thyroxine (75 microg eltroxine/ 100 g(-1) body weight, ip for 7 days). Treatment increased PA activity significantly in rat kidneys. No changes were seen in PA, PAI and glucose in plasma of rats. There was significant decrease in total cholesterol and LDL-cholesterol levels in serum of treated group resulting in the decrease of HDL/LDL and total cholesterol/cholesterol ratios. However, triglycerides and VLDL showed significant higher activity in the serum of treated group as compared to controls. The results suggest beneficial effects of thyroxine treatment by increasing PA activity in kidney and reducing the cholesterol content in blood. It may be helpful to prevent hypercoagulable state by maintaining the normal homeostatic balance and restoring renal function.

Brothers of women with polycystic ovary syndrome: a new group with high risk for type 2 diabetes mellitus and cardiovascular disease

Polycystic ovary syndrome [PCOS] is common endocrinal disorder which is highly inherited and characterized by many metabolic abnormalities. We hypothesized that male relatives of PCOS women would also have metabolic abnormalities. Thus, our aim was to assess insulin sensitivity and metabolic parameters in brothers of women with PCOS and male control individuals. 30 brothers of women with PCOS and 20 male healthy control subjects were included in the study. Brothers and control were subjected to complete medical evaluation with stress on anthropometric measurements, fasting insulin, homeostasis assessment model [HOMA-IR], lipids, plasminogen activator inhibitor-1 [PAI-1], C-reactive protein [CRP] and androgens. Brothers and control individuals were similar as regard to age, MBI, WHR and blood pressure. However, brothers were insulin resistant and had dyslipidemia and dyscoagulability [HOMA-IR, P=0.043, TC P=0.001, LDL-C P=0.002, HDL-C P=0.03, TG P=0.048, PAI-1 P=0.002, CRP P=0.046]. Also HOMA-IR, was correlated significantly with BMI p<0.001, WHR P<0.001, PAI-1 P<0.001, CRP P<0.01, TG, P<0.001, LDL-C P=0.02, HDL-C P=0.019]. Brothers of women with PCOS have a metabolic phenotype consisting of dyslipidemia, insulin resistance, dysmgulability and carry an increased risk of cardiovascular disease [CVD] and type 2 diabetes mellitus [type 2 DM]. Given the high prevalence of PCOS, brothers may represent an important new risk factor for CVD in men and should be considered a well identified group for primary preventive measures

Renin angiotensin aldosterone system and fibrinolytic activity in experimentally induced diabetic and hypertensive rats

Many evidences pointed to certain relations between renin angiotensin aldosterone system [RAAS] and endogenous fibrinolytic system. To examine the effect of low salt supplement, as an activator to endogenous RAAS on plasma fibrinolytic function in alloxan diabetic L-NAME induced hypertensive ncphritic rats in the presence or absence of angiotensin converting enzyme inhibitor [ACEI] and/or aldosterone antagonist. Diabetes was induced in male Wistar rats by a single intraperitoneal [i.p] injection of alloxan [150 mg/kg]. Rats were received low salt supplement [0.08% NaCI] and N[G]-nitro-L-arginine methyl ester [L-NAME, 0.1 mg/ml] in the drinking water during the experimental periods. Treatment of diabetic hypertensive nephritic rats with ACEI [mocxipril hydrochloride, 7 mg/kg body weight daily and orally], aldosterone antagonist [spiranolactone, 25 mg/kg body weight daily and orally] or their combined administration for 6 weeks. Glucose, creatinine, sodium, potassium, aldosterone, ACE activity, transforming growth factor-beta 1 [TGF-beta] were determined in serum, whereas, renin, plasminogen activator inhibitors [PAI-1 were estimated in plasma. In urine nitric oxide [NO] concentration was evaluated and total. DNA and RNA were recorded in kidney tissues. During low salt intake, activation of the RAAS was monitored through observed significant Increase in serum alciosterone, sodium, potassium, and ACE activity. Impairment of renal function following diabetes and L-NAME administration was manifested By increase in serum glucose, mean arterial pressure [MAP] heart rate [HR]. serum creatinine and low urinary NO level, these data demonstrated that activation of RAAS in diabetic hypertensive nephritic rats significantly increased PAI-1 activity, TGF-beta1 and dry thrombus weight. It markedly decreased total DNA contents in kidney homogenate. Interruption of the RAAS with the ACEI, aldosterone antagonist or their combined administration lot 6 weeks significantly decreased PAl-1 activity TGF-beta1 and thrombus weight [fibrinolytic actvity]. However urinary NO, kidney content of total DNA showed significant increase. Combined form therapy has a better effect regarding PAI-1 TGF-beta 1 and NO than monotherapy. Data obtained provides an evidence of direct functional association between the RAAS and the fibrinolytic system in rats. This may help to elucidate possible mechanisms by which ACE inhibition and aldosterone antagonist exerts antagonist exerts vasculoprotective effects and reduce the risk of renal atherothromhotic events closely related to uncontrolled diabetes

Study on the function of decoction for invigorating the kidney and improving blood circulation to thrombosis on rabbit blood stasis model / 中国中药杂志

<p><b>OBJECTIVE</b>To evaluate the effect of decoction for invigorating the kidney and improving blood circulation to thrombosis and pathology on rabbit blood stasis model.</p><p><b>METHOD</b>Thirty rabbits were ramdomly divided into normal group, model group, high dose group, low dose group and Xue Shuan Ning group. Tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor (PAI), fibrinogen (Fbg) and D-dimer (DD) were investigated after those rabbits had been treated. One rot was solected randomly from each group to observe pathological changes.</p><p><b>RESULT</b>There were significant differences in t-PA, PAI, Fbg and DD between normal group and other groups is very obvious (P < 0.01) . Between groups of high dose low dose Xue Shuan Ning and model, the statistical differeces were significant, as well as between groups of high dose, low dose and Xue Shuan Ning groups (P < 0.05). However, there was no statistical difference between high dose group and high dose group (P > 0.05). The pathological changes in model group were most serious, those in Xue Shuan Ning were less serious. There were slight pathological changes in high dose group and low dose group.</p><p><b>CONCLUSION</b>Models ware made successfully. High dose group and low dose group have stronger effect on thrombosis than Xue Shuan Ning group.</p>

Effects of chronic mercury poisoning on blood coagulation and fibrinolysis systems / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the effects of chronic mercury poisoning on blood coagulation and fibrinolysis systems, and the possible mechanism.</p><p><b>METHODS</b>Twenty-seven patients with chronic mercury poisoning were studied with 30 healthy people as control. Thrombomodulin (TM), tissue plasminogen activator (t-PA), plasminogen activator inhibitor (PAI), interleukin-13 (IL-13), interleukin-18 (IL-18), soluble intercellular adhesion molecule-1 (SICAM-1) were examined with ELISA methods, and superoxide dismutase (SOD) and lipid peroxidation (LPO) was examined with chemical catalysis methods. Two to three weeks after treatment with reduced glutathione, tiopronin and daidzein, blood was used for determin the above items again.</p><p><b>RESULTS</b>(1) The concentration of TM in patients [(2.36 +/- 0.16) ng/ml] was significantly lower than in the control [(4.36 +/- 0.24) ng/ml] (P < 0.01), while TM tended to be higher after treatment [(4.82 +/- 0.34) ng/ml] (P < 0.05). (2) The concentration of t-PA in patients [(3.44 +/- 0.34) ng/ml] was significantly lower than in the control [(4.52 +/- 0.16) ng/ml] (P < 0.05), and was higher significantly [(5.63 +/- 0.58) ng/ml] after treatment (P < 0.05); The concentration of PAI in patients [(48.23 +/- 3.59) ng/ml] was significantly higher than in the control [(31.59 +/- 2.13) ng/ml] (P < 0.05), but after treatment no significant change [(50.71 +/- 4.29) ng/ml] was found (P > 0.05). (3) The activity of SOD in patients [(953.85 +/- 9.56) U/g Hb] was significantly lower than in the control [(1,308.75 +/- 10.21) U/g Hb] (P < 0.01), and was higher significantly [(1,217.95 +/- 6.29) U/g Hb] after treatment (P < 0.05); and the concentration of LPO in patients [(9.53 +/- 0.26) nmol/ml] was significantly higher than in the control (P < 0.05), and significantly lower [(7.29 +/- 0.35) nmol/ml] after treatment (P < 0.05). (4) The concentrations of IL-13 [(35.93 +/- 5.28) pg/ml], IL-18 [(28.79 +/- 2.53) pg/ml], SICAM-1 [(603.16 +/- 29.12) ng/ml] were significantly higher than those in the controls (P < 0.05, P < 0.01), but no significant difference was found after treatment.</p><p><b>CONCLUSION</b>Dysfunction of the TM/protein C system and t-PA/PAI system (i.e. the decrease of anti-coagulation activity and the inhibition of the function for the fibrolysis system) may play a key role in the secondary hypercoagulable state induced by chronic mercury poisoning.</p>

Role of adhesion molecules in vascular complications of type 2 diabetes mellitus

The present study was undertaken to evaluate the plasma levels of soluble adhesion molecules [intercellular adhesion molecule-1 [ICAM-1] and vascular cell adhesion molecule-1 [VCAM-l] in type 2 DM to elucidate its potential involvement in pathogenesis of diabetic vascular complications and its association with other independent risk factors for diabetic atherosclerosis. The study was conducted on 60 patients with type 2 DM [27 with vascular complications Vs. 33 without vascular complications, assessed by fundus examinations] and 20 healthy controls, at Al-Azhar University Hospitals between March 2004 to August 2004. Serum levels of ICAM-l and VCAM-l in association with fasting blood sugar, fasting insulin and insulin resistance were measured, as wall as lipid profile, plasminogen activator inhibitor [PAl] and factor VII. Serum levels of ICAM-1 and VCAM-1 were significantly higher in type 2 diabetic patients with vascular complication than those without vascular complications, but no significant changes in their levels were found between patients without vascular complications and controls. Serum levels of ICAM- 1 and VCAM- I were significantly correlated with other studied parameters in patients with and without vascular complications except fasting serum insulin levels. Serum levels of insulin resistance [IR] and lipid profile were significantly higher in patients with type 2 DM than controls and in patients with vascular complications than those without vascular complications. Serum levels of fasting blood glucose, factor VII and PAl were significantly higher in diabetic patients with and without vascular complications than controls, on the other hand no significant changes was found between patients with and those without vascular complications. The present study suggested that the levels of soluble adhesion molecules in type 2 DM with dyslipidaemia, hyperinsulinemia and hypercoagulable state may be a marker of endothelial cell activation or dysfunction and may be related to the activity of multiple cell types in atherosclerotic lesion. Serum levels of ICAMs were closely related to vascular diabetic complications. Furthermore, they may serve as a tool for monitoring the impact of prevention and intervention on vascular damage

Impaired systemic fibrinolysis in severe neonatal respiratory distress syndrom

The intravascular and intra-alveolar deposition of fibrin in severe neonatal respiratory distress syndrome [RDS] have been attributed to activation of clotting. We questioned whether in face of enhanced clotting, fibrinolysis is sufficient in these neonates. Therefore, we aimed to assess plasminogen activator inhibitor-1 [PAI-1] as a marker of fibrinolysis in plasma of neonates with RDS to investigate its relation to disease severity and the possible prognostic value of its early measurement. The study included 65 neonates, all of them were clinically assessed within 6 hours of birth for inclusion in the study. The study group consisted of 45 preterm neonates, 30 with RDS, and 15 healthy preterm control neonates. The remaining twenty neonates were fullterms; 10 had clinical evidence of infection and 10 were healthy fullterm control neonates. Neonates with RDS were clinically and radiologically evaluated to assign severity and accordingly they were categorized into a severe group and a mild to moderate group. Blood samples were obtained from these neonates while on mechanical ventilation during the first day of life. Fraction of inspiratory oxygen [FiO2] was determined and they were clinically followed up throughout the whole duration of ventilation. Laboratory investigations included CBC, C-reactive protein [CRP], ABG and plasma PAI-1 determination for all neonates as well as a coagulation assay including PT, PTT, fibrinogen and fibrin degradation products [FDPS] done for 18 RDS neonates. Mean plasma concentrations of PAI-1 were significantly elevated in PT neonates with RDS as a whole [P<0.001] and in each subgroup [P<0.01] as compared to control PT neonates. Severe RDS group showed significantly higher PAI-1 in plasma as compared to the mild to moderate group [P<0.001]. A significant positive correlation existed between PAI-1 and FiO2 in all neonates with RDS [P<0.01]. Fibrinogen levels were significantly lower in neonates with severe RDS as compared to the mild to moderate group [P<0.05] and they were negatively correlated to plasma PAI-1 in all studied RDS neonates [P<0.05]. The ratio of FDPs between 5 and 20 to FDPs<5 was 9:1 in the mild to moderate RDS group, compared to 1:1 in the severe group. Plasma levels of PAI-1 in full term neonates with systemic infection were significantly higher as compared to healthy fullterms [P<0.05]. Sepsis was documented in 21.05% of deaths among neonates with RDS. Retrospective tracing of CRP in neonates with RDS who died revealed values that were non-significantly higher than those of survivors [P>0.05]. Meanwhile, Plasma PAI-1 levels in deceased were significantly higher than those of survivors [P<0.001]. PAI-1 cut off value of 58 ng/ml was 60% sensitive and 73.68% specific to predict mortality in RDS. In severe RDS, high PAI-1 impairs systemic fibrinolysis which likely facilitates the deleterious effects of early clotting activation, contributing to disease severity and mortality. Plasma level of PAI-1 within 24 hours of birth, though might be influenced by early infection may be a useful predictor of outcome in neonatal RDS

Study on function of decoction for invigorating the kidney and improving blood circulation on rabbits blood stasis model / 中国中药杂志

<p><b>OBJECTIVE</b>To evaluate the effect of decoction for invigorating the kidney and improving blood circulation to thrombosis on rabbits blood stasis model.</p><p><b>METHOD</b>Thirty rabbits were randomly divided into normal group, model group, heavy dose group, slight dose group and xue shuan ning group. Tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor (PAI), fibrinogen (Fbg) and D-dimer (DD) were investigated after those rabbits had been treated. One was selected randomly from each group to observe pathological changes.</p><p><b>RESULT</b>There was significant difference in t-PA, PAI, Fbg and DD between normal group and other groups (P < 0.01). Among groups of heavy dose, slight dose, xue shuan ning and model, the statistical differences were significant, as well as among groups of heavy dose, slight dose and xue shuan ning (P < 0.05). However, there was no statistical difference between heavy dose group and slight dose group (P > 0.05). The pathological changes in model group were most serious, and those in xue shuan ning were less serious. There were slight pathological change in heavy dose group and light dose group.</p><p><b>CONCLUSION</b>Models were made successfully. Heavy dose group and slight dose group have stronger effect on thrombosis than xue shuan ning group.</p>

Curative machanism of Shenle capsule on 5/6 nephrectomy rats / 中国中药杂志

<p><b>OBJECTIVE</b>To explore curative machanism of Shenle capsule on the 5/6 nephrectomy rats.</p><p><b>METHOD</b>Fibrin plate method was applied to examine activity of urinary plasminogen activator(PA). Semi-quantitative analysis was used to observe stained intensity and area of tissue-type plasminogen activator, urokinas-type plasminogen activator/ plasminogen activator inhibitor(tPA, uPA/PAI-1)in remnant renal tissue. Northern blot was employed to analyze the expression of transforming growth factor (TGF-beta) mRNA.</p><p><b>RESULT</b>In model control group, the urinary PA activity and protein expression of tPA, uPA were down-regulated, but protein expression of PAI-1, TGF-beta mRNA was up-regulated in remnant renal tissue. In each treated group, the urinary PA activity and protein expression of tPA/uPA were enhanced,but the protein expression of PAI-1, TGF-beta mRNA decreased simultaneously.</p><p><b>CONCLUSION</b>Shenle capsule can delay glomerulosclersis and tubulointerstitial fibrotic lesions of remnant kidney by improving the activity of urinary PA and modulating the expression of tPA, uPA/PAI-1 and TGF-beta mRNA.</p>

Parâmetros hemostáticos na gravidez normal e após o parto / Hemostatic parameters in normal pregnancy and postpartum

Relatos da literatura revelam que a gravidez normal está associada a complexas alterações da hemostasia que resultam em um estado de hipercoagulabilidade sanguínea. O objetivo do presente estudo foi estabelecer a evolução de 8 marcadores que refletem diversas etapas do processo hemostático, avaliados ao final da gravidez e 4 meses após o parto. A condição hemostática de 8 gestantes que não tiveram complicações durante a gravidez e não apresentavam doenças intercorrentes no 3º trimestre de gestação (Grupo I) e 4 meses após o parto (Grupo II) foi avaliada através da determinação dos níveis plasmáticos de trombomodulina (TM), ativador tissular do plasminogênio (t-PA), inibidor do ativador do plasminogênio (PAI), fragmento 1 + 2 da protrombina (F¹+²), plasminogênio (Plg), fibrinogênio, Dímero D (D-Di) e da contagem de plaquetas. A análise dos resultados mostrou uma diferença significativa entre as médias obtidas nos dois grupos para TM (p<0,05), t-PA (p<0,01), PAI-1 (p<0,05), fibrinogênio (p<0,01),F ¹+² (p<0,001) e D-Di (p< 0,001). Para plasminogênio e contagem de plaquetas nenhuma diferença foi observada. Estes resultados permitem concluir que a gravidez normal está associada a uma exacerbação do mecanismo da coagulação e que o sistema fibrinolítico está funcionando adequadamente, o que previne a formação de trombos. Quatro meses após o parto, nenhuma alteração nos parâmetros hemostáticos avaliados foi observada, caracterizando o retorno à condição hemostática normal.

Relationship between plasminogen activator, plasminogen activator inhibitor and Sertoli cells / 中华男科学杂志

Plasminogen activator(PA) and plasminogen activator inhibitor(PAI) are involved in many physiological or pathological events. The Sertoli cells, the important elements within the seminiferous epithelium, are thought to play a key role in spermatogenesis. The Sertoli cells secrete PA and PAI. The levels of them are modulated by hormonal and cell-mediated influences. They play a fundamental role in the maintenance of spermatogenesis, sperm motility and fertilization.

Effect of growth hormone on fibrinolytic system in rat.

Growth hormone deficiency (GHD) is associated as a risk factor in increased mortality from cardiovascular diseases. Abnormal lipid profile and increased levels of plasminogen activator inhibitor (PAI) and fibrinogen have been noted in GHD patients. Present study was carried out to investigate the effect of growth hormone (GH) on plasminogen activator (PA) activity in heart, levels of PA, PAI, glucose and fibrinogen in plasma and serum lipid profile. Rats were injected 125 mU GH kg-1 body weight subcutaneously daily for one week. PA activity was significantly higher in the heart of GH treated rats as compared to controls. GH treatment decreased plasma glucose and fibrinogen levels significantly. No significant differences were seen in PA, PAI in plasma, triglycerides and total cholesterol in serum of the two groups of rats. A significant increase in high density lipoprotein cholesterol (HDL) occurred in GH treated group resulting into a decrease in LDL/HDL ratio. The results indicate that GH may be beneficial in cardiovascular diseases as it decreases the levels of plasma fibrinogen and increases the level of HDL in blood and also increases the level of PA in heart.

Evaluación retrospectiva del uso de activador tisular del plasminógeno (rt-PA) en el manejo del infarto agudo de miocardio en el servicio de urgencias de la Clínica Shaio

Objetivo: Evaluar el uso de activador tisular del plasminógeno recombinante (rt-PA) en los pacientes con IAM que ingresaron por el servicio de urgencias. Materiales y métodos: Estudio descriptivo retrospectivo de corte transversal en donde se tomaron todos los pacientes con diagnóstico de IAM al ingreso en urgencias durante el período comprendido entre enero de 1996 hasta julio de 1997 en los cuales se utilizó la terapia trombolítica con rt-PA. Se administró el esquema acelerado de rt-PA seguido por infusión de heparina para mantener PTT entre 50 y 70 segundos por 24 a 48 horas. Las indicaciones para administrar rt-PA fueron: edad menor de 75 años, infarto de localización anterior y tiempo de evolución menor de 6 horas. La recolección de la información se realizó a partir de la historia clínica. Las variables estudiadas fueron edad, sexo, tipo de infarto, localización, clasificación de killip, tiempo de evolución al momento de la consulta, uso, tipo y causa de exclusión de trombosis con rt-AP. permeabilidad de la arteria relacionada con el infarto, realización de angioplastia, requerimiento de cirugía (electiva o de urgencia), complicaciones y mortalidad antes de la intervención con angioplastia electiva o cirugía. Resultados: De un total de 458 pacientes con diagnóstico de IAM se realizó terapia de recanalización en 206 pacientes (44.5 por ciento), 12 con angioplastia primaria (6 por ciento), 146 (70.5 por ciento) con STK y 48 (23.5 por ciento) con rt-PA. De estos pacientes 42 (87.5 por ciento) fueron hombres y 6 (12.5 por ciento) mujeres. Las edades estuvieron entre 42 años el más joven y 75 años el de mayor edad, con un promedio de 60-17 años. El tiempo promedio de consulta a urgencias después del inicio de los síntomas fue de 4.9 horas. Ningún paciente entró con Killip IV, 3 (6.5 por ciento) Killip I, 14 (29 por ciento) en Killip II yb 31 (64.5 por ciento) en Killip III. La principal causa de exclusión de trombólisis con rt-PA fue la consulta fuera de tiempo > 6 horas. Se presentó un caso de hemorragia de vías digestivas clasificada como menor ya que no requirió transfusión. De las complicaciones secundarias al IAM se presentó choque en 4 (8.3 por ciento), arritmia ventricular en 3 (6.2 por ciento); pericarditis en 2 (4.15 por ciento); angina post IAM en 1 (2.5 por ciento); insuficiencia mitral en 1 (2.5 por ciento); trombo intracavitario 1 (2.5 por ciento). La mortalidad entre los pacientes trombolizados...

Terapêutica de reposiçäo hormonal / Hormone replacement therapy

A mulher, no período pré-menopausa, está relativamente protegida de doença cardiovascular. Admite-se que essa proteção seja devida ao estrogênio e a seus efeitos sobre fatores de risco e mecanismos de aterogênese. A terapêutica de reposição hormonal, definida como tratamento com estrogênio ou a combinação estrogênio e protestógeno, representa, hoje, tema de permanente debate, considerando que sua indicação pode modificar o impacto de várias doenças que afetam a mulher na menopausa. A terapêutica de reposição pode reduzir o risco de osteoporose, câncer de cólon, doença de Alzheimer e doença isquêmica miocárdica, e, simultaneamente, aumentar a mortalidade devida a câncer de mama e de endométrio. Até que estejam disponíveis os resultados de ensaios clínicos aleatórios, com "endpoints" adequadamente definidos de doença cardiovascular, a decisão para o uso de reposição hormonal deverá ser individualizada, levando em conta os possíveis benefícios em mulheres de alto risco cardiovascular frente aos riscos de câncer de endométrio e câncer de mama.

Clot lysis: role of plasminogen activator inhibitors in haemostasis and therapy.

An unimpeded circulation of blood depends on the concerted activation of coagulation and fibrinolytic factors. The latter entails the controlled, localised conversion of plasma zymogen plasminogen to the active enzyme plasmin mediated by tissue-type plasminogen activator (tPA). Bulk of tPA activity is in the proximity of the endogenous plasminogen activator inhibitor (PAI) as an active complex. The advent of molecular biology techniques has enabled isolation of cDNA for the inhibitors PAI-1, PAI-2 and PAI-3 and data indicate that these belong to the serine protease inhibitor (Serpine) family with arginine as its active site but immunologically distinct from each other. Enhanced tPA or PAI-1 forms one of the risk factors related to cardiac diseases and thrombotic disorders. A line of therapy entails lowering of PAIs with concomitant increase in tPA levels leading to net enhancement in fibrinolytic activity. In as much as plasminogen activators exert their action extracellularly, they are accessible to inhibitors and therefore PAIs could have a therapeutic potential and serve as prognostic indicators in cancer. Documented findings related to the biochemical characteristics and therapeutic potential of PAIs are presented and discussed in the review.

The Significance of Plasma Urokinase-type Plasminogen Activator and Type 1 Plasminogen Activator Inhibitor in Lung Cancer / 결핵

BACKGROUND: Cancer invasion and metastasis require the dissolution of the extracellular matrix in which several proteolytic enzymes are Involved. One of these enzymes is the urokinase - type plasminogen activator(u-PA), and plasminogen activator inhibitors(PAI-1, PAI-2) a]so have a possible role in cancer invasion and metastasis by protection of cancer itself from proteolysis by u-PA. It has been reported that the love]s of u-PA and plasminogen activator inhibitors in various cancer tissues are significantly higher than those in normal tissues and have significant correlations with tumor size and lymph node involvement Here, we measured the concentration of plasma u-PA and PAI- 1 antigens in the patients with lung cancer and compared the concentration of them with histologic types and staging parameters. METHODS: We measured the concentration of plasma u-PA and PAI-1 antigens using commercial ELISA kit in 37 lung cancer patients, 21 benign lung disease patients and 24 age-matched healthy controls, and we compared the concentration of them with histologic types and staging parameters in lung cancer patients. RESULTS: The concentration of u-PA was 1.0α0.3ng/mL in controls, 1.0α0.3ng/mL in benign lung disease patients and 0.9α0.3ng/mL in lung cancer patients. The concentration of PAI-1 was 14.2α6.7ng/mL in controls, 14.9α6.3ng/mL in benign lung disease patients, and 22.1 α9.8ng/mL in lung cancer patients. The concentration of PAI- 1 in lung cancer patients was higher than those of benign lung disease patients and controls. The concentration of u-PA was 0.7α0.4ng/mL in squamous cell carcinoma, 0.8α 0.3ng/mL in adenocarcinoma, 0.9ng/mL in large cell carcinoma, and 1.1α0.7ng/mL in small cell carcinoma. The concert traction of PAI-1 was 22.3α7.2ng/mL in squamous cell carcinoma, 22.6α9.9ng/mL in adenocarcinoma, 42ng/mL in large cell carcinoma, and 16.0α14.2ng/mL in small cell carcinoma. The concentration of u-PA was 0.74ng/mL in stage I, 1.2α0.6ng/mL in stage II, 0.7 α 0.4ng/mL in stage IIIA, 0.7α0.4ng/mL in stage IIIB, and 0.7α0.3ng/mL in stage IV. The concentration of PAI-1 was 21.8ng/mL in stage I, 22.7α8.7ng/mL in stage II, 18.4 α4.9ng/mL in stage IIIA, 25.3α9.0ng/mL in stage IIIB, and 21.5α10.8ng /mL in stage IV. When we divided T stage unto T1-3 and 74, the concentration of u-PA was 0.8α 0.4ng/mL in T1-3 and 0.7α0.4ng/mL in T4, and the concentration of PAI-1 was 17.9α 5.6ng/mL in T1-3 and 26.1α9.1ng/mL in T4. The concentration of PAI-1 in T4 was significantly higher than that in T1-3. The concentration of u-PA was 0.8α 0.4ng/mL in M0 and 0.7α0.3ng/mL in Ml, and the concentration of PAI-1 was 23.6α8.3ng/mL in M0 and 21.5α10.8ng/mL in M1 CONCLUSIONS: The plasma levels of PAI-1 in lung cancer were higher than benign lung disease and control, and the plasma levels of PAI-1 in 74 were significantly higher than T1-3. These findings suggest involvement of PAI-1 with local invasion of lung cancer, but it should be confirmed by the data on comparison with pathological staging and tissue level in lung cancer.

Tissue-type plasminogen activator, plasminogen activator. Inhibitor in stressed newborns

Stressed newborns have an increased prevalence of thrombotic disease. Estimation of tissue-type plasminogen activator [tPA] antigen and activity, plasminogen activator inhibitor [PAI] antigen and activity, fibrinogen level, prothrombin time [PT] and partial thromboplastin time [PTT] were determined in fifteen stressed and fifteen normal neonates. The stressed group showed significantly higher levels of [tPA] antigen and activity. Also [PAI] antigen and activity were significantly elevated in stressed neonates. On the other hand fibrinogen concentration, showed in significant decrease in stressed neonates. Prothrombin time and partial thromboplastin time showed insignificant prolongation in stressed babies. This study proved that stressed neonates increase [tPA] and activate lytic system. However, the activity is suboptimal due to associated increase in [PAI]

Epidemiología de los estados trombofílicos / Epidemiology of the thrombophilic states

Esta revisión hace referencia a la epidemiología de los estados trombofílicos más importantes, con todas las dificultades para establecer su verdadera prevalencia, la cual se halla influenciada por diversos factores: demográficos, diagnósticos, poblacionales, entre otros. De los pacientes con antecedentes trombóticos recurrentes y/o historia familiar, sólo un 10 por ciento obedece a una trombofilia, existiendo alrededor de un 25 por ciento que posiblemente la presente, pero en donde su etiología no ha podido ser determinada con los métodos hoy disponibles. Las alteraciones fibrinolíticas continúan siendo objeto de discusión, con respecto a la relevancia clínica y a su carácter hereditario