Plasminogen Research Articles

Blood coagulation biomarkers for postoperative venous thromboembolism diagnosis in orthopedic traumatic patients: a case control study

Objective To evaluate the value of blood coagulation biomarkers in orthopaedic traumatic patients after surgery and analyze its diagnostic values for venous thrombosis embolism. Methods In thiscase control study, we consecutive enrolled 108 traumatic patients after surgery. 54 patients have thrombosis and other 54 patients have no thrombosis. Blood was taken 3-4 days after surgery. Routine coagulation screening test , FDP(fibrinogen/fibrin degradation products) , D dimer and new item such as TM(thrombomodulin), TAT(thrombin-anti-thrombin complex), t-PAIC(tissue- type plasminogen activator-plasminogen activator inhibitor complex), PIC(plasmin-anti-plasmin complex) were tested. The difference between groups of these biomarkers was compared, and then the receiver operation curve (ROC) was drew to determine the diagnostic cut-off point and diagnostic performance. Results ALL blood coagulation biomarkers in orthopaedic traumatic patients after surgery were significantly increased. The group of patients with thrombosis have higher TM(9.04±2.06)IU/ml, t-PAIC(10.15±4.23)ng/ml, PIC(1.15±0.70)μg/ml, D dimer(5.31±5.10)ng/ml than group without thrombosis TM(7.50±1.70)IU/ml, t-PAIC (6.97±2.56)ng/ml, PIC(0.93±0.84)μg/ml, D dimer(2.35±2.12)ng/ml, and P=0.000 2, <0.000 1, <0.000 1, <0.000 1, respectively. However, TAT(4.79±4.32)ng/ml, (6.51±5.92)ng/ml, FDP(8.87±7.68)μg/ml, (4.91±4.67)μg/ml showed no difference between thrombosis groupand no thrombosis group, (P=0.212 3, 0.050 8; respectively). The area under the ROC curve of TM, t-PAIC, PIC and D-dimer were 0.718 5, 0.741 6, 0.648 0, 0.670 0, respectively; P values were <0.000 1, <0.000 1, 0.009 3, 0.004 1, respectively; cut-off values were 11.15 IU/ml, 10.65 ng/ml, 1.36 μg/ml, 7.69 ng/ml, respectively; positive likelihood ratios were 9.00, 11.29, 3.66, 14.60, respectively; specificity were 98.15%, 96.23%, 90.20%, 97.96%, respectively; the diagnostic rates were 20.3%, 46.3%, 35.8%, 25.9%, respectively. Conclusions There were coagulation and fibrinolysis system activated in orthopaedic traumatic patients after surgery.TM, t-PAIC, PIC, D dimer were good biomarkers for the diagnosis of thrombosis after trauma surgery. TAT was not fit for screening thrombosis after surgery because of influence of anti-coagulation.(Chin J Lab Med, 2016, 39: 751-755) Key words: Postoperative complications; Venous thrombosis; Biological markers; Thrombomodulin; Tissue plasminogen activator; Fibrin fibrinogen degradation products

Effects of recombinant adenovirus with high expression of osteopontin gene on malignant biological behaviors of hepatocellular carcinoma cells

Objective To study the effect of recombinant adenovirus with high expression of osteopontin (OPN) gene on malignant biological behaviors of hepatocellular carcinoma (HCC) cells. Methods HCC (HepG2) cells were cultured and divided into rAd-OPN group and rad NC group. HCC cells in rAd-OPN group were transfected with recombinant adenovirus with high expression of OPN gene, and thsose in NC group were transfected with recombinant adenovirus containing invalid fragment. The expression of OPN, and cell migration, invasion and metastasis associated genes was detected. Results At 12, 24, 36, and 48 h after adenovirus transfection, mRNA expression of OPN in rAd-OPN group was 1.93±0.25, 2.58±0.30, 3.61±0.57 and 3.88±0.63, cell migration rate was (18.20±2.76)%, (43.76±7.17)%, (62.48±9.25)% and (77.33±10.24)%, and transmembrane cell number was (25.24±4.26), (32.13±4.97), (40.27±6.24) and (47.95±7.25) cells respectively, significantly higher than in rAd-NC group (P<0.05). At 48 h after transfection of adenovirus, mRNA expression of matrix metalloproteinase (MMP)-2, MMP-9, urokinase-type plasminogen activator (uPA) in rAd-OPN group was respectively 2.32±0.49, 1.78±0.28, 3.14±0.51, significantly higher than that in rAd NC group (P<0.05). Conclusion Recombinant adenovirus with high expression of OPN gene can promote the migration, invasion and up-regulate the metastasis-related gene expression. Key words: Carcinoma, hepatocellular; Osteopontin; Migration; Invasion

Airway and peripheral uPAR is elevated in asthma, and identifies a severe, non-atopic subset of patients

Rationale: Genetic polymorphisms in the asthma susceptibility gene, urokinase plasminogen activator receptor (uPAR/PLAUR) have been associated with lung function decline and uPAR blood levels in asthma subjects. Preliminary studies have identified uPAR elevation in asthma; however, a definitive study regarding which clinical features of asthma uPAR may be driving is currently lacking. Objectives: We aimed to comprehensively determine the uPAR expression profile in asthma and control subjects utilizing bronchial biopsies and serum, and to relate uPAR expression to asthma clinical features. Methods: uPAR levels were determined in control (n = 9) and asthmatic (n = 27) bronchial biopsies using immunohistochemistry, with a semi-quantitative score defining intensity in multiple cell types. Soluble-cleaved (sc) uPAR levels were determined in serum through ELISA in UK (cases n = 129; controls n = 39) and Dutch (cases n = 514; controls n = 96) cohorts. Measurements and main results: In bronchial tissue, uPAR was elevated in inflammatory cells in the lamina propria (P = 0.0019), bronchial epithelial (P = 0.0002) and airway smooth muscle cells (P = 0.0352) of patients with asthma, with uPAR levels correlated between the cell types. No correlation with disease severity or asthma clinical features was identified. scuPAR serum levels were elevated in patients with asthma (1.5-fold; P = 0.0008), and we identified an association between high uPAR serum levels and severe, nonatopic disease. Conclusions: This study provides novel data that elevated airway and blood uPAR is a feature of asthma and that blood uPAR is particularly related to severe, nonatopic asthma. The findings warrant further investigation and may provide a therapeutic opportunity for this refractory population.

Use of IV TPA for Eligible Stroke Patients Continues to Increase

In the more than 20 years since intravenous tissue plasminogen activator (IV TPA) was shown to improve outcomes for acute ischemic stroke, the process

Acute Antithrombotics and Hemorrhage After Ischemic Stroke

For patients with acute ischemic stroke who receive tissue plasminogen activator (TPA), antithrombotic agents such as aspirin customarily are withheld

Libby amphibole-induced mesothelial cell autoantibodies bind to surface plasminogen and alter collagen matrix remodeling.

Lamellar pleural thickening (LPT) is a fibrotic disease induced by exposure to Libby amphibole (LA) asbestos that causes widespread scarring around the lung, resulting in deterioration of pulmonary function. Investigating the effects of autoantibodies to mesothelial cells (MCAA) present in the study populations has been a major part of the effort to understand the mechanism of pathogenesis. It has been shown in vitro that human mesothelial cells (Met5a) exposed to MCAA increase collagen deposition into the extracellular matrix (ECM). In this study, we sought to further elucidate how MCAA drive increased collagen deposition by identifying the protein targets bound by MCAA on the cellular surface using biotinylation to label and isolate surface proteins. Isolated surface protein fractions were identified as containing MCAA targets using ELISA. The fractions that demonstrated binding by MCAA were then analyzed by tandem mass spectrometry (MS/MS) and MASCOT analysis. The most promising result from the MASCOT analysis, plasminogen (PLG), was tested for MCAA binding using purified human PLG in an ELISA. We report that serum containing MCAA bound at an optical density (OD) 3 times greater than that of controls, and LA‐exposed subjects had a high frequency of positive tests for anti‐PLG autoantibodies. This work implicates the involvement of the plasminogen/plasmin system in the mechanism of excess collagen deposition in Met5a cells exposed to MCAA. Elucidating this mechanism could contribute to the understanding of LPT.

SuPAR remains uninfluenced by surgery in septic patients with bloodstream infection.

Surgical trauma induces activation of the immune system and may cause an increase of inflammatory biomarkers tested postoperatively in septic patients treated for bloodstream infection. The aim of this study was to determine the impact of surgical interventions on the novel sepsis biomarker soluble urokinase plasminogen activator receptor (suPAR) and to compare results with those of routine laboratory parameters CRP, PCT, and IL-6 in patients with culture-proven bloodstream infection. Forty-six adult patients with positive blood culture undergoing minor or major surgical intervention were investigated, 12 blood culture positive patients served as control group. Blood was collected 24 hours before and after surgical intervention for determination of the sepsis biomarkers suPAR, CRP, PCT, and IL-6. Within the surgical study cohort, a non-significant increase of suPAR, CRP, and PCT was observed postoperatively (p 0.642; p 0.773; p 0.087). In contrast, a slight decrease of IL-6 (p 0.599) was observed. A significant correlation was calculated for the pre- and postoperative difference of CRP (p 0.028) and PCT (p 0.008) and type of surgical intervention received: after minor surgical intervention only PCT decreased significantly (p<0.001), while after major surgical interventions no significant differences were observed for all biomarkers evaluated. In the control group, a significant decrease of CRP (p 0.005) and PCT (p 0.005) was observed. In patients treated adequately for bloodstream infections, postoperative suPAR levels remained uninfluenced of the surgical trauma and might therefore be a reliable parameter for postoperative infectious monitoring. After minor surgical intervention, PCT seems to be the most reliable parameter.

Analysis of Plasminogen Genetic Variants in Multiple Sclerosis Patients.

Multiple sclerosis (MS) is a prevalent neurological disease of complex etiology. Here, we describe the characterization of a multi-incident MS family that nominated a rare missense variant (p.G420D) in plasminogen (PLG) as a putative genetic risk factor for MS. Genotyping of PLG p.G420D (rs139071351) in 2160 MS patients, and 886 controls from Canada, identified 10 additional probands, two sporadic patients and one control with the variant. Segregation in families harboring the rs139071351 variant, identified p.G420D in 26 out of 30 family members diagnosed with MS, 14 unaffected parents, and 12 out of 30 family members not diagnosed with disease. Despite considerably reduced penetrance, linkage analysis supports cosegregation of PLG p.G420D and disease. Genotyping of PLG p.G420D in 14446 patients, and 8797 controls from Canada, France, Spain, Germany, Belgium, and Austria failed to identify significant association with disease (P = 0.117), despite an overall higher prevalence in patients (OR = 1.32; 95% CI = 0.93–1.87). To assess whether additional rare variants have an effect on MS risk, we sequenced PLG in 293 probands, and genotyped all rare variants in cases and controls. This analysis identified nine rare missense variants, and although three of them were exclusively observed in MS patients, segregation does not support pathogenicity. PLG is a plausible biological candidate for MS owing to its involvement in immune system response, blood-brain barrier permeability, and myelin degradation. Moreover, components of its activation cascade have been shown to present increased activity or expression in MS patients compared to controls; further studies are needed to clarify whether PLG is involved in MS susceptibility.

Soluble urokinase type plasminogen activator receptor as a predictor of chronic kidney disease

Soluble urokinase type plasminogen activator receptor as a predictor of chronic kidney disease

Is Low-Dose Thrombolysis for Acute Ischemic Stroke a Viable Option?

Epidemiologic studies have shown that Asians have an increased risk for hemorrhagic stroke compared with white patients. This pattern has led to concerns about the safety of full-dose IV tissue plasminogen activator (TPA) in Asian patients with acute ischemic stroke. In Japan, a reduced dose of TPA (0.6 mg/kg) has been approved instead of the conventional dose (0.9 mg/kg). The current study was a randomized trial of these two doses of TPA in …

Phenotype and genotype analysis of in herited dysplasminogenemia caused by plasminogen Ala601Thr mutation

Objective To detectthe phenotype and gene mutations underlying aninherited dysplasminogenemia pedigree and search the virulence gene. Methods The peripheral venous blood samples of the proband and his family members(fourteen subjects of three generations in total)were collected, and their prothrombin time(PT), activated partial thromboplastin time(APTT), thrombin time(TT), fibrinogen(FIB), fibrinogen degradation products(FDP), D-dimmer(D-D)weretested on a STAGO analyzer, the plasminogen activity(PLG: A)and plasminogen antigen(PLG: Ag)were analyzedby thechromogenic substrate assay and rocket immunoelectrophoresis, respectively. All 19 exons, 5' and 3' untranslated regions of PLG were amplified with PCR. Direct DNA sequencing was used to analyze the amplified products, which were confirmed by backward sequencing. Three bioinformatics online softwares(SIFT, PolyPhen-2 and MutationTaster)were used to forecast the possible impact of the mutations on the protein function. At last, the model analysis of mutate site was taken on a Swiss-Pdb Viewer software. Results The PLG: Avalue of the proband and other 6 family members were decreased to the half, while the PLG: Ag was normal. The D-D and FDP value of the proband, his grandma and father were slightly higher. DNA sequencing has revealed that the proband and the other 6 members of this family had the same mutation of g. 38829G>A in exon 15, leading to the missense mutationp.Ala601Thr.The results of bioinformatics softwares showed that the mutation could affect the thePLGfunction. Protein model analysis indicated that the hydrophobic interaction force and hydrogen bond between the amino acids were changed, which might affect the stability of the PLG.In addition, all the members of this family take the heterozygous SNP of g. 2501C>A in the 5'UTR. Conclusions The p. Ala601Thr found in the inherited dysplasminogenemia pedigree in the exon 15 was responsible for the reduced PLG: A of the family, the dysplasminogenemia and this mutation were both reported for the first time in China.(Chin J Lab Med, 2016, 39: 366-371) Key words: Mutation, Missense; Plasminogen; Polymor phism, single nucleotide

Thrombolysis of plasmin-ΔK on photochemically induced branch retinal vein occlusion after intravitreal injection in rats

Background Retinal vein occlusion is a common retinal vascular diseases.Thromblysis and anticoagulation therapies are main approaches.However, systemic thrombolysis is relatively inefficient, and it often enhances the risk of hemorrhage. Objective This study was to investigate the therapeutic effects of PLM-ΔK, a kringle deficiency mutant of plasmin, on photochemically induced branch retinal vein occlusion (BRVO) after intravitreal injection. Methods BRVO models were established by the combination of caudal vein injection of Rose Bengal with argon laser radiation of periphery area of retinal veins in SD rats.Forty model rats were randomized into balance salt solution (BSS) group and 0.01 U, 0.02 U, 0.03 U PLM-ΔK group, and 10 μl corresponding drug was intravtreally injected 12 hours after modeling.Ophthalmoscopy and fundus fluorescein angiography (FFA) were performed to observe the change of retinal veins.The animals were sacrificed 3 days after intravitreal injection, and hematoxylin and eosin staining was used for the histopathological and ultrastructural examination of retinas.The retina of the rats was isolated for the stretched preparation of retina.The expressions of fibronectin (FN) and laminin (LN) in eyeball wall were assayed by immunofluorescence technology.The use and care of the animals complied with Statement of the Association for Research in Vision and Ophthalmology. Results The revascularization of over 2 retinal veins was found in 0, 3, 6 and 8 rats in the BBS group and 0.01 U, 0.02 U, 0.03 U PLM-ΔK group 3 days after intravitreal injection, respectively, showing a significant difference among the groups (χ2=9.635, P=0.022), and the rat number with revascularization in 0.01 U PLM-ΔK group was not significantly different from that in BSS group (Z=-1.558, P=0.119), but the difference between 0.03 U PLM-ΔK group and 0.01 U PLM-ΔK group was significant (Z=-2.762, P=0.006). In the third day after intravitreal injection, retinal vein thrombus were found in the BSS group under the light microscope, and angiogenesis was seen on the retinal flatmount nuclear.In the 0.03 U PLM-ΔK group, posterior vitreal detachment was exhibited under the light microcope, and no retinal new vessel and cell damage were seen.FN was strongly expressed in the inner limiting membrane (ILM) layer, photocyte layer, outer limiting membrane (OLM) layer, choroid and scleral layer, and LN was expressed mainly in the ILM, OLM and scleral layer in the BSS group.However, the expression intensities of FN and LN were obviously weakened in the 0.03 U PLM-ΔK group. Conclusions Intravitreal injection of PLM-ΔK can enhance the reperfusion of occluded branch retinal vein and serve as a potential therapeutic drug for BRVO.Also it can permeate into choroid after intravitreal injection to degradate FN and LN. Key words: Retinal vein occlusion/therapy; Fibrinolytic agents/administration d Intravitreal injection; Thrombolytic therapy/methods; Disease models, animal; SD rats

Evaluation of two novel leptospiral proteins for their interaction with human host components.

Pathogenic species of the genus Leptospira are the etiological agents of leptospirosis, the most widespread zoonosis. Mechanisms involved in leptospiral pathogenesis are not well understood. By data mining the genome sequences of Leptospira interrogans we have identified two proteins predicted to be surface exposed, LIC10821 and LIC10064. Immunofluorescence and proteinase K assays confirmed that the proteins are exposed. Reactivity of the recombinant proteins with human sera has shown that rLIC10821, but not rLIC10064, is recognized by antibodies in confirmed leptospirosis serum samples, suggesting its expression during infection. The rLIC10821 was able to bind laminin, in a dose-dependent fashion, and was called Lsa37 (leptospiral surface adhesin of 37 kDa). Studies with human plasma components demonstrated that rLIC10821 interacts with plasminogen (PLG) and fibrinogen (Fg). The binding of Lsa37 with PLG generates plasmin when PLG activator was added. Fibrin clotting reduction was observed in a thrombin-catalyzed reaction, when Fg was incubated with Lsa37, suggesting that this protein may interfere in the coagulation cascade during the disease. Although LIC10064 protein is more abundant than the corresponding Lsa37, binding activity with all the components tested was not detected. Thus, Lsa37 is a novel versatile adhesin that may mediate Leptospira-host interactions.

Glyceraldehyde 3-phosphate dehydrogenase and galectin from Dirofilaria immitis participate in heartworm disease endarteritis via plasminogen/plasmin system

The interaction between parasitic protozoa and helminths, both in the blood and in tissues and the fibrinolytic system of their hosts is usually considered as a survival parasite mechanism since this system is the physiological route responsible for degrading fibrin clots. The broad-range proteolytic activity of plasmin, the final enzyme of the route, implies that its recruitment by these parasites is an important mechanism that mediates their invasion and establishment in the hosts. However, recent studies have proposed a dual role for plasmin by linking its over-production with pathological mechanisms at vascular level. Most of these studies have been conducted in Dirofilaria immitis, a blood-borne parasite that survives in the pulmonary arteries of its host for years while it produces a chronic inflammatory disease, whose main pathogenic mechanism is the appearance of proliferative endarteritis. Recently, the participation of two proteins from D. immitis, glyceraldehyde 3-phosphate dehydrogenase (DiGAPDH) and galectin (DiGAL), in the activation of the fibrinolytic system of its host has been demonstrated, which has been a priori associated with parasite survival mechanisms. The aim of the present paper was to study the role of plasmin generated by these proteins in the emergence of proliferative endarteritis. An in vitro model of canine endothelial and smooth muscle cells, as well as the two parasitic recombinant proteins were employed. The results show that DiGAPDH and DiGAL stimulate the proliferation and migration of both cell types, as well as the degradation of the extracellular matrix (ECM) via plasminogen (PLG)/plasmin system, being all of these mechanisms related to the appearance of proliferative endarteritis. Due to the high degree of evolutionary conservation of these antigens, these data support the hypothesis of the survival/pathology ambivalence in the interactions between parasites and the fibrinolytic system of their hosts and represent an advance in the knowledge of the mechanisms involved in the emergence of proliferative endarteritis in the cardiopulmonary dirofilariosis (heartworm disease).

Relationship of uric acid levels with clinical outcomes in severe ischemic stroke patients treated with intravenous recombinant tissue plasminogen activator

One hundred and seventy six consecutive patients with acute ischemic stroke who received intravenous recombinant tissue plasminogen activator(rt-PA) in 4.5 hours from symptom onset during February 2009 to July 2013 were included in the study. Modified Rankin Scale was used to evaluate the recovery of neurological functions. Patients were divided into good (0-1) or poor (2-6) outcome groups according modified Rankin Scale score. Univariate analysis and multivariate logistic regression analysis were used to determine the differences of clinical data between the two groups. The age of patients with good outcome was significantly lower than that of poor outcome group [(61.4±11.5) vs. (69.0±13.2) years, P=0.000]. Compared to patients with poor outcomes, patients with good outcome group showed lower rate of diabetes[ 13%(12/93) vs. 29%(24/83), P=0.009], lower blood glucose level [(5.05±0.97) vs. (5.83±1.72) mmol/L, P=0.020], higher uric acid level[(404.4±151.7) vs. (345.6±107.5) μmol/L, P=0.028], shorter onset to treatment time [(1.92±0.94) vs. (2.30±1.01) h, P=0.019], lower baseline National Institute of Health Stroke Scale score [(14.0±5.2) vs. (16.0±6.2), P=0.025], lower systolic blood pressure level at 2 h[(140.8±18.3) vs.(149.0±18.9) mmHg(1 mmHg=0.133 kPa), P=0.005]and 24 h [(137.6±21.9) vs. (147.1±17.4) mmHg, P=0.009]after thrombolysis. Logistic regression analysis showed that uric acid levels were not related to hemorrhagic transformation independently (P=0.172, OR=0.965, 95%CI: 0.917-1.016), but were related to outcome independently (P=0.047, OR=0.957, 95%CI: 0.916-0.999). Key words: Uric acid, blood; Tissue plasminogen activator, recombinant; Stroke, ischemic

Localization and identification of thrombin and plasminogen activator activities in model human thrombi by in situ zymography.

Human thrombi vary in their susceptibility to lysis and this is clinically important. Several potential contributory factors were examined in this study by using model thrombi, created under flow; these provide a robust, reproducible and easily-manipulated system. Here we identify the plasminogen activators (PA) active in model thrombi of known age and define the cellular and plasma contribution to activity in different areas. The cell-rich head of model thrombi had strong thrombin and PA activity, with coagulant activity also at the tail. Thrombin activity decreased as model thrombi were aged. PA activity in the thrombus head also decreased on ageing of thrombi but activity emerged around the thrombi, including the tail. Activity in the head of fresh model thrombi was primarily due to uPA, with some contribution from tPA. Experiments with thrombi prepared from platelet-rich plasma and added leucocytes showed that uPA activity at the head of fresh thrombi was derived from PMN. Older thrombi had tPA activity around the tail of the thrombus; this activity occurred in the absence of cells. This study highlights the importance of PMN-derived uPA activity in the lysis of fresh thrombi, with activity originating in the leucocyte-rich head. It also shows that thrombi are dynamic structures in which fibrin can be repeatedly laid down and lysed, observations that are relevant to therapeutic lysis and potential rethrombosis.

Change in endogeous hydrogen sulfide in patients with acute pancreatitis and its relationship to coagulation function

To study the change in endogenous hydrogen sulfide (H2S) in patients with acute pancreatitis and its relationship to coagulation function. A prospective case control study was conducted. Forty patients with mild acute pancreatitis (MAP group) and 40 with severe acute pancreatitis (SAP group) admitted to Yiwu Central Hospital in Zhejiang Province from December 2002 to March 2015 were enrolled. Forty healthy persons served as control (healthy control group). Blood was collected to determine the levels of H2S, blood coagulation factor Ⅷ (FⅧ), von Willebrand factor (vWF), plasminogen (PLG), antithrombin (AT), platelet count (PLT), tissue factor (TF), tumor necrosis factor-α (TNF-α), and protease activated receptor-1 (PAR-1). The correlations among the above parameters were analyzed. There was no statistical significance in sex, age, body weight and time of disease among three groups, indicating it was comparable among the groups. Compared with healthy control group, the levels of H2S, FⅧ, vWF, TF, TNF-α, and PAR-1 in MAP and SAP groups were significantly elevated [H2S (μmol/L): 67.42±6.34, 112.47±12.69 vs. 42.57±4.18, FⅧ: (67.5±5.8)%, (82.3±4.7)% vs. (57.2±6.4)%, vWF: (112.6±9.7)%, (142.5±12.5)% vs. (76.4±8.2)%, TF (ng/L): 45.27±4.34, 64.76±6.25 vs. 18.15±1.89, TNF-α (ng/L): 197.67±13.62, 324.72±25.54 vs. 20.08±2.57, PAR-1 (fluorescence intensity): 32.16±4.43, 56.12±7.07 vs. 12.27±2.12, all P < 0.01], and PLG and AT activity were significantly decreased [PLG: (52.4±4.7)%, (36.7±3.2)% vs. (62.1±5.6)%, AT: (43.2±6.9)%, (35.5±5.4)% vs. (53.6±6.1)%, all P < 0.01]. The changes in the parameters in SAP group were more remarkable than those in MAP group (all P < 0.01). PLT in SAP group was significantly lower than that in healthy control and MAP groups (×109/L: 8.5±1.1 vs. 15.7±2.8, 12.4±1.9, both P < 0.01). H2S was positively correlated with FⅧ, vWF, TF, TNF-α, and PAR-1 (r value was 0.56, 0.61, 0.72, 0.66, 0.64, respectively, all P < 0.01), and it was negatively correlated with PLG and AT (r value was -0.64, -0.57, both P < 0.01). As an inflammatory factor, endogenous H2S deteriorates coagulation function in patients with acute pancreatitis by up-regulating TF, TNF-α, and PAR-1.

Efficacy and safety of rt-PA intravenous thrombolysis for treating acute ischemic stroke beyond the therapeutic window: a Meta-analysis

Objective To evaluate the efficacy and safety of thrombolytic therapy with recombinant tissue-type plasminogen activator (rt-PA) in the treatment of acute ischemic stroke with broadened therapeutic window (4.50-6.00 h). Methods Online databases, such as PubMed, Cochrane Library, China National Knowledge Infrastructure (CNKI), Wanfang data were searched from January 1980 to August 2015 with key words: acute ischemic stroke, thrombolytic therapy, beyond 4.5 hours. The included studies were evaluated according to Jadad Scale, and Meta-analysis was performed by using RevMan 5.3 software. Results There were 681 relevant records through preliminary searching, and 3 randomized controlled trials (RCTs) were finally included, involving 2033 patients with acute ischemic stroke (1029 patients in rt-PA thrombolytic therapy and 1004 patients in placebo therapy), who were treated within 6 h from the onset of symptoms. Results of Meta-analysis showed there were no statistical differences between rt-PA group and placebo group on modified Rankin Scale (mRS) score ( RR = 1.070, 95%CI: 0.940-1.220; P = 0.310), Barthel Index (BI) score ( RR = 1.040, 95%CI: 0.940-1.160; P = 0.430) and mortality ( RR = 1.260, 95%CI: 0.990-1.610; P = 0.060). However, the incidence of intracerebral hemorrhage (ICH) in rt-PA group was significantly higher than that in placebo group ( RR = 1.550, 95% CI: 1.030-2.340; P = 0.030). Conclusions Thrombolytic therapy with rt-PA in broadened therapeutic window (4.50-6.00 h) is not effective for treating acute ischemic stroke, and the risk of ICH is increased with rt-PA treatment. However, this conclusion still needs to be verified with more high-quality, multi-center, large-sample RCTs. DOI: 10.3969/j.issn.1672-6731.2016.02.002

Effect of TGP capsules combined with urokinase for the patients with acute cerebral ischemic stroke

Objective To evaluated the effect of Total Glucosides of Paeony (TGP)combined with urokinase for the patients with acute cerebral ischemic stroke. Methods A total of 152 patients with ACIS were divided into the control group (77 patients) and the observation group (75 patients) by completely randomized grouping method. Patients in the control group were treated with Urokinase by intravenous, and the observation group were treated with TGP combined with Urokinase. All patients were treated with thrombolysis treatment with in the 24 h after the exclusion of cerebral hemorrhage by brain CT and coagulation mechanism disorder. In the following 14 days, they were given the treatment of anti platelet aggregation, lowering blood lipid, trophic nerve and improving circulation. The National Institutes of Health Stroke Scale (NIHSS), the modified Rankin scale(mRS) and the daily life ability index (Barthel) were measured. Results There were 28 (37.3%) patients with the NIHSS scores decreasing 4 points in observation group and 12 (15.6%) in control group after treatment of 7 days, and there was significant difference between two groups (χ2=8.181, P=0.004). There were 68 (90.7%) patients with mRS score less than 4 points in observation group and 51 (15.6%) in control group after treatment of 14 days, and there was significant difference between two groups (χ2=11.945, P=0.001). The NIHSS scores in the observation group was lower than control group (8.3 ± 2.2 vs. 9.9 ± 2.5; t=4.192, P<0.05). The mRS score in the observation group was lower than control group (2.3 ± 0.9 vs. 2.9 ± 1.1; t=3.684, P<0.01), and the Barthel index in the observation group was higher than the control group (88.7 ± 16.2 vs. 77.5 ± 15.2; t=4.401, P<0.01). Conclusions The TGP capsules combined with Urokinase intravenous thrombolysis could improve the neurological deficit, and the ability of daily life of ACIS patients. Key words: Brain infarction; Acute disease; Urokinase-type plasminogen activator; TGP capsules; Integrated Chinese traditional and western medicine therapy

Group A Streptococcus exploits human plasminogen for bacterial translocation across epithelial barrier via tricellular tight junctions

Group A Streptococcus (GAS) is a human-specific pathogen responsible for local suppurative and life-threatening invasive systemic diseases. Interaction of GAS with human plasminogen (PLG) is a salient characteristic for promoting their systemic dissemination. In the present study, a serotype M28 strain was found predominantly localized in tricellular tight junctions of epithelial cells cultured in the presence of PLG. Several lines of evidence indicated that interaction of PLG with tricellulin, a major component of tricellular tight junctions, is crucial for bacterial localization. A site-directed mutagenesis approach revealed that lysine residues at positions 217 and 252 within the extracellular loop of tricellulin play important roles in PLG-binding activity. Additionally, we demonstrated that PLG functions as a molecular bridge between tricellulin and streptococcal surface enolase (SEN). The wild type strain efficiently translocated across the epithelial monolayer, accompanied by cleavage of transmembrane junctional proteins. In contrast, amino acid substitutions in the PLG-binding motif of SEN markedly compromised those activities. Notably, the interaction of PLG with SEN was dependent on PLG species specificity, which influenced the efficiency of bacterial penetration. Our findings provide insight into the mechanism by which GAS exploits host PLG for acceleration of bacterial invasion into deeper tissues via tricellular tight junctions.