The Effect of the Early Application of Tirofiban on Acute Ischemic Stroke (AIS) after Intravenous Thrombolysis with Urokinase

Objective:Discussion and analysis of the effect of the early application of Tirofiban on acute ischemic stroke(AIS)after intravenous thrombolysis with urokinase.Method:The subjects of this study are 40 patients with AIS admitted at the Yibin Fourth People’s Hospital,of which were computer-randomized into a control group(20 cases,with regular urokinase intravenous thrombolysis therapy)and a research group(20 cases,combined with early Tirofiban treatment)from January 2018 to December 2022.The intervention outcomes between these two groups were compared and analyzed.Result:The blood platelet-related parameters before treatment had no statistical difference between the two groups(P>0.05),but the research group was higher than that of the control group after treatment(P<0.05).The Barthel index before treatment in both groups had no statistical difference(P>0.05),but the research group was higher than that of the control group after treatment(P<0.05).Conclusion:Early Tirofiban treatment for patients with AIS after intravenous thrombolysis with urokinase could effectively regulate the blood platelet-related parameters,hence improving treatment benefits and living capacity for patients,with definite clinical benefits.

Human urokinase-type plasminogen activator gene-modifiedbone marrow-derived mesenchymal stem cells attenuateliver fibrosis in rats by down-regulating the Wnt signalingpathway

AIM: To evaluate the therapeutic effects of bone marrow-derived mesenchymal stem cells(BMSCs) with human urokinase-type plasminogen activator(u PA) on liver fibrosis, and to investigate the mechanism of gene therapy.METHODS: BMSCs transfected with adenovirusmediated human urokinase plasminogen activator(Adu PA) were transplanted into rats with CCl4-induced liver fibrosis. All rats were sacrificed after 8 wk, and their serum and liver tissue were collected for biochemical, histopathologic, and molecular analyzes. The degree of liver fibrosis was assessed by hematoxylin and eosin or Masson's staining. Western blot and quantitative reverse transcription-polymerase chain reaction were used to determine protein and m RNA expression levels.RESULTS: Serum levels of alanine aminotransferase, aminotransferase, total bilirubin, hyaluronic acid, laminin, and procollagen type Ⅲ were markedly decreased, whereas the levels of serum albumin were increased by u PA gene modified BMSCs treatment. Histopathology revealed that chronic CCl4-treatment resulted in significant fibrosis while u PA gene modified BMSCs treatment significantly reversed fibrosis. By quantitatively analysing the fibrosis area of liver tissue using Masson staining in different groups of animals, we found that model animals with CCl4-induced liver fibrosis had the largest fibrotic area(16.69% ± 1.30%), while fibrotic area was significantly decreased by BMSCs treatment(12.38% ± 2.27%) and was further reduced by u PA-BMSCs treatment(8.31% ± 1.21%). Both protein and m RNA expression of β-catenin, Wnt4 and Wnt5 a was down-regulated in liver tissues following u PA gene modified BMSCs treatment when compared with the model animals.CONCLUSION: Transplantation of u PA gene modified BMSCs suppressed liver fibrosis and ameliorated liver function and may be a new approach to treating liver fibrosis. Furthermore, treatment with u PA gene modified BMSCs also resulted in a decrease in expression of molecules of the Wnt signaling pathway.

Clinical outcomes of transcatheter selective superior mesenteric artery urokinase infusion therapy vs transjugular intrahepatic portosystemic shunt in patients with cirrhosis and acute portal vein thrombosis

AIM To compare the outcomes of transcatheter superior mesenteric artery(SMA) urokinase infusion and transjugular intrahepatic portosystemic shunt(TIPS) for acute portal vein thrombosis(PVT) in cirrhosis.METHODS From January 2013 to December 2014, patients with liver cirrhosis and acute symptomatic PVT who met the inclusion criteria were randomly assigned to either an SMA group or a TIPS group. The two groups accepted transcatheter selective SMA urokinase infusion therapyand TIPS, respectively. The total follow-up time was24 mo. The primary outcome measure was the change in portal vein patency status which was evaluated by angio-computed tomography or Doppler ultrasound.Secondary outcomes were rebleeding and hepatic encephalopathy.RESULTS A total of 40 patients were enrolled, with 20 assigned to the SMA group and 20 to the TIPS group. The symptoms of all patients in the two groups improved within 48 h. PVT was improved in 17(85%) patients in the SMA group and 14(70%) patients in the TIPS group. The main portal vein(MPV) thrombosis was significantly reduced in both groups(P < 0.001), and there was no significant difference between them(P= 0.304). In the SMA group, superior mesenteric vein(SMV) thrombosis and splenic vein(SV) thrombosis were significantly reduced(P = 0.048 and P = 0.02),which did not occur in the TIPS group. At 6-, 12-,and 24-mo follow-up, in the SMA group and the TIPS group, the cumulative rates free of the first episode of rebleeding were 80%, 65%, and 45% vs 90%, 80%,and 60%, respectively(P = 0.320); the cumulative rates free of the first episode of hepatic encephalopathy were 85%, 80%, and 65% vs 50%, 40%, and 35%,respectively(P = 0.022).CONCLUSION Transcatheter selective SMA urokinase infusion and TIPS are safe and effective for acute symptomatic PVT in cirrhosis.

Urokinase-type plasminogen activator is a modulator of synaptic plasticity in the central nervous system:implications for neurorepair in the ischemic brain

The last two decades have witnessed a rapid decrease in mortality due to acute cerebral ischemia that paradoxically has led to a rapid increase in the number of patients that survive an acute ischemic stroke with various degrees of disability.Unfortunately,the lack of an effective therapeutic strategy to promote neurological recovery among stroke survivors has led to a rapidly growing population of disabled patients.Thus,understanding the mechanisms of neurorepair in the ischemic brain is a priority with wide scientific,social and economic implications.Cerebral ischemia has a harmful effect on synaptic structure associated with the development of functional impairment.In agreement with these observations,experimental evidence indicates that synaptic repair underlies the recovery of neurological function following an ischemic stroke.Furthermore,it has become evident that synaptic plasticity is crucial not only during development and learning,but also for synaptic repair after an ischemic insult.The plasminogen activating system is assembled by a cascade of enzymes and their inhibitors initially thought to be solely involved in the generation of plasmin.However,recent work has shown that in the brain this system has an important function regulating the development of synaptic plasticity via mechanisms that not always require plasmin generation.Urokinase-type plasminogen activator(uPA)is a serine proteinase and one of the plasminogen activators,that upon binding to its receptor(uPAR)not only catalyzes the conversion of plasminogen into plasmin on the cell surface,but also activates cell signaling pathways that promote cell migration,proliferation and survival.The role of uPA is the brain is not fully understood.However,it has been reported while uPA and uPAR are abundantly found in the developing central nervous system,in the mature brain their expression is restricted to a limited group of cells.Remarkably,following an ischemic injury to the mature brain the expression of uPA and uPAR increases to levels comparable to those observed during development.More specifically,neurons release uPA during the recovery phase from an ischemic injury,and astrocytes,axonal boutons and dendritic spines recruit uPAR to their plasma membrane.Here we will review recent evidence indicating that binding of uPA to uPAR promotes the repair of synapses damaged by an ischemic injury,with the resultant recovery of neurological function.Furthermore,we will discuss data indicating that treatment with recombinant uPA is a potential therapeutic strategy to promote neurological recovery among ischemic stroke survivors.

MEASUREMENT AND CORRELATION OF PARTITION COEFFICIENTS OF UROKINASE IN AQUEOUS TWO-PHASE PEG-POTASSIUM PHOSPHATE SYSTEM

Partition coefficients of Urokinase(UK)were measured in aqueous two-phase systems con-taining polyethylene glycol and potassium phosphate at 273.2K.Based on the Diamond-Hsu model,a modified expression was obtained for the correlation of enzyme partitioning in theabove-mentioned systems.Utilizing a modified form,the partitioning data of UK and heteroproteinwere correlated.The results show that the modified model is simple gives good precision.and will fa-cilitate engineering scale-up of aqueous two-phase systems for certain proteins purification.

Matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 expression in early focal cerebral infarction following urokinase thrombolysis in rats

Activity of matrix metalloproteinase-9 increases following cerebral ischemia/reperfusion, and is associated with cerebral microvascular permeability, blood-brain barrier destruction, inflammatory cell infiltration and brain edema. Matrix metalloproteinase-9 also likely participates in thrombolysis. A rat model of middle cerebral artery infarction was established by injecting autologous blood clots into the internal carotid artery. At 3 hours following model induction, urokinase was injected into the caudal vein. Decreased neurological severity score, reduced infarct volume, and increased expression of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 were observed in the cerebral cortex 24 hours after urokinase thrombolysis. These results suggest that urokinase can suppress damage in the acute-early stage of cerebral infarction.

Gene expression changes of urokinase plasminogen activator and urokinase receptor in rat testes at postnatal stages

Aim： To investigate the gene expression changes of urokinase plasminogen activator （uPA）/urokinase receptor （uPAR） in rat testes at postnatal stages and explore the effects of uPA/uPAR system on the rat spermatogenesis. Methods： The mRNAs of uPA and uPAR in rat testes were measured by using real-time quantitative polymerase chain reaction （PCR） at postnatal days 0, 5, 10, 15, 21, 28, 35, 42, 49 and 56, respectively. Results： The tendencies of uPA and uPAR mRNA expression were similar at most postnatal stages except for Do. The expression of uPAR mRNA in rats testes was relatively higher than that of uPA at postnatal Do, and both were decreased until D21, increased obviously at postnatal D28, reached a peak at postnatal D35, then declined sharply at postnatal D42 and retained at a low level afterwards. Conclusion： The uPA/uPAR system may be strongly linked to spermiation and spermatogenesis via regulating germ cell migration and proliferation, as well as promoting the spermiation and detached residual bodies from the mature spermatids.

Studies on the Relationship between Urokinase Plasminogen Activator(uPA)and Human Sperm Motility

To clarify the role of urokinase plasminogen activator(uPA) in the mechanisms of regulating sperm motility and the ability of fertilizing, we investigated the quantities and activities of uPA in human seminal Plasma and on the membrane of spermatozoa.Semens were harvested from 22 infertile patients with asthenospermia and 20 healthy fertile men according to WHO standards. To quantify the membrane-bound uPA in the samples, polyclonal antibodies against human urokinase were employed by means of a sandwich ELISA. The uPA activities in seminal plasma and on the surface of spermatozoa were determined using Agarose-Fibrine-Plate method and the experiment of immunological identification with polyclonal antibodies against urokinase. In lysates of spermatozoa, significantly lower levels of uPA(23. 1±7.35 mu/106 cells ) and uPA activity (5.13±3.85 mu/106 cells) were found in patient group as compared to healthy fertile men exhibiting normospermia (29. 89±9. 40 mu/105 cells and 10. 17±6. 18 mu/106 cells). In seminal plasma, uPA activity in patient group (2134±1581. 3 IU/L)was also found significantly lower than that of normal group (3365±1859. 5 IU/L). Positive correlations were observed between sperm motility and uPA quantities (r=0. 48, P＜0. 005), as well as with uPA activities (r= 0. 45,P＜0. 005).Thus, it is inferred that membrane associated uPA on human spermatozoa may be related directly to sperm motility and fertility.

AFFINITY CHROMATOGRAPHY PURIFICATION OF UROKINASE WITH EPICHLOROHYDRIN ACTIVATED AGAROSE MATRIX

1 INTRODUCTIONIn literature,most matrices of affinity chromatography for urokinase(EC 3.4.99.26)purification were prepared by cyanogen bromide activation.However,theseadsorbents usually suffered from the drawback of leakage of the ligand,particularly inalkaline medium,because of the instability of the isourea linkage between the ligandand the spacer or agarose.Moreover,the positively charged imido group of theN-substituted isourea derivative and the hydrophobicity of the spacers might promotenonspecific adsorption.On the contrary,the adsorbents prepared by the method

Genetic association of urokinase-type plasminogen activator gene rs2227564 site polymorphism with sporadic Alzheimer's disease in the Han Chinese population

A missense C/T polymorphism in exon 6 （the NCBI rslD is rs2227564） of the urokinase-type plasminogen activator gene has been identified as a possible hot spot for Alzheimer＇s disease risk. The present study analyzed urokinase-type plasminogen gene polymorphisms of rs2227564 with sporadic Alzheimer＇s disease by PCR-restriction fragment length polymorphism. Results showed that CC, CT and TT genotype distribution frequencies had significant differences between sporadic Aizheimer＇s disease patients and healthy controls, in-depth analysis of the association between urokinase-type plasminogen gene rs2227564 polymorphisms and sporadic Alzheimer＇s disease indicated that people with the C-positive genotype CC ＋ CT were at a higher risk for developing sporadic Alzheimer＇s disease. These results support the contribution of the polymorphisms of rs2227564 in the urokinase-type plasminogen gene to the pathogenesis of sporadicAIzheimer＇s disease in the Han Chinese population.

Sub-Tenon's urokinase injection-assisted vitrectomy in early treatment of suprachoroidal hemorrhage: Four cases report

BACKGROUND Suprachoroidal hemorrhage(SCH) is a rare but potentially catastrophic ocular event. Surgery for SCH is often challenging because of the difficulty in resolving the retinal and choroidal detachment. Here, we describe a novel surgical technique in which urokinase is administered by sub-Tenon's injection to target an organized clot in SCH prior to drainage.CASE SUMMARY A consecutive case series of four eyes with serous and hemorrhagic choroidal detachments secondary to cataract surgery or trauma was documented to evaluate the feasibility of using a sub-Tenon's urokinase injection-assisted 23-gauge and 20-gauge incision to drain choroidal detachments. Urokinase(2000 IU) was given by sub-Tenon's injection one day before surgery for clot liquefaction. A 23-gauge infusion line was placed in the anterior chamber. A 20-gauge incision was created in the suprachoroidal space 3.5 mm from the limbus. After drainage, pars plana vitrectomy was performed because of concomitant pathology that demanded this additional procedure. Visual acuity, ocular findings, the timing of surgical interventions, surgical procedures, and outcomes were retrospectively reviewed in four patients. Postoperative follow-up of the patients ranged from 6 to 24 mo(mean, 13 mo). After the treatment, all patients achieved excellent anatomical recovery. CONCLUSION Sub-Tenon's urokinase injection-assisted vitrectomy makes clot liquefaction happen in the early treatment stage, resulting in marked stability during the procedure.

UROKINASE THROMBOLYSIS IN ARTERY FOR TREATMENT OF PERIPHERAL ARTERIAL OCCLUSION

Purpose:To evaluate the clinical efficacyof thrombolysis injecting urokinase inperipheral artery for treatment of arterialocclusion.Materials and methods:12 patientswere treated with injecting urokinase inartery.Results:The mean dose of urokinase was500000U,the mean time of thrombolysis was200 minutes.The rate of clinic efficacy was83%.Conclusion:The indication of urokinasethrombolysis in artery is for those patientswith arterial occlusion in whom catheteriza-tion is feasible and clinical course is inone month.This operation is simple and conv-enient,it does not require special equipment.The method can be used safely and effectivelyin patients with arterial occlusion.

Influence of irbesartan combined with low dose urokinase on treatment effect and serologic indexes in elderlyisolated systolic hypertension patients

To study the influence of irbesartan combined with low dose urokinase on treatment effect and serologic indexes in elderly isolated systolic hypertension patients. Methods: One hundred and ten cases of elderly isolated systolic hypertension patients in our hospital during January 2013–January 2016 were randomly divided into observation group with 55 cases and control group with 55 cases. Patients in observation group received irbesartan combined with low dose hydrochlorothiazide treatment and those in control group received amlodipine combined with low dose hydrochlorothiazide treatment, each lasted for 4 weeks. After 4 weeks treatment, non-invasive blood pressure and blood pressure variability indexes were measured, serum endothelial function, inflammatory factors and oxidative stress indexes were measured. Results: After 4 weeks treatment, patients systolic blood pressure (SBP), 24 h mean systolic blood pressure (24 h SBP), 24 h systolic blood pressure standard deviation (24 h SSD) were significantly lower in observation group than in control group (P<0.05);serum endothelial function index EF-1 was lower in observation group than in control group, while NO and endothelial nitric oxide synthase (eNOS) were higher in observation group than in control group (P<0.05);serum high sensitivity C-reactive protein (hs-CRP), interleukin (IL)-1β, IL-6, IL-8, tumor necrosis factor-α (TNF-α) contents were lower in observation group than in control group (P<0.05);serum oxidation index such as malondialdehyde (MDA) was lower in observation group than in control group, while antioxidant indexes such as total anti-oxidative capacity (TAC), superoxide dismutase (SOD), GSH were higher in observation group than in control group (P<0.05). Conclusions: Irbesartan combined with low dose hydrochlorothiazide can effectively reduce systolic hypertension and blood pressure variability in elderly isolated systolic hypertension patients, while optimizing the systemic inflammation and oxidative stress status.

Effect of urokinase in combined with minimally invasive hematoma puncture with YL-1 type needle on the blood sugar and serum CRP in patients with hypertensive cerebral hemorrhage

Objective:To observe the clinical efficacy of urokinase in combined with minimally invasive hematoma puncture with YL-1 type needle in the treatment of hypertensive cerebral hemorrhage and the effect on blood sugar and serum CRP.Methods:A total of 84 patients with hypertensive cerebral hemorrhage who were admitted in our hospital were included in the study and divided into the minimally invasive group (n=53) and the conservative group (n=31) according to different treatment protocols. The patients in the two groups were given routine drug treatments. The patients in the observation group were given urokinase in combined with minimally invasive hematoma puncture with YL-1 type needle. The blood sugar and serum CRP levels before and after treatment in the two groups were compared. CT was performed to reexamine the cerebral hematoma and edema volume.Results: The serum CRP and blood sugar levels 3, 7 and 14 d after treatment in the minimally invasive group were significantly lower than those in the conservative group (P<0.05). The cerebral hematoma and edema volume 1, 3, 7, and 14 d after treatment in the minimally invasive group was significantly lower than that in the conservative group (P<0.05).Conclusions: Urokinase in combined with minimally invasive hematoma puncture with YL-1 type needle in the treatment of hypertensive cerebral hemorrhage can significantly alleviate the brain tissue injury, reduce the systemic inflammatory reaction and blood sugar level, and contribute to the rehabilitation.

Intracoronary arterial retrograde thrombolysis with percutaneous coronary intervention: a novel use of thrombolytic to treat acute ST-segment elevation myocardial infarction

Background Clearance of coronary arterial thrombosis is necessary in patients with acute ST-segment elevation myocardial infarction (STEMI) undergoing urgent percutaneous coronary intervention (PCI). There is currently no highly-recommended method of thrombus removal during interventional procedures. We describe a new method for opening culprit vessels to treat STEMI: intracoronary arterial retrograde thrombolysis (ICART) with PCI. Methods & Results Eight patients underwent ICART. The guidewire was advanced to the distal coronary artery through the occlusion lesion. Then, we inserted a microcatheter into the distal end of the occluded coronary artery over the guidewire. Urokinase (5–10 wu) mixed with contrast agents was slowly injected into the occluded section of the coronary artery through the microcatheter. The intracoronary thrombus gradually dissolved in 3–17 min, and the effect of thrombolysis was visible in real time. Stents were then implanted according to the characteristics of the recanalized culprit lesion to achieve full revascularization. One patient experienced premature ventricular contraction during vascular revascularization, and no malignant arrhythmias were seen in any patient. No reflow or slow flow was not observed post PCI. Thrombolysis in myocardial infarction flow grade and myocardial blush grade post-primary PCI was 3 in all eight patients. No patients experienced bleeding or stroke. Conclusions ICART was accurate and effective for treating intracoronary thrombi in patients with STEMI in this preliminary study. ICART was an effective, feasible, and simple approach to the management of STEMI, and no intraprocedural complications occurred in any of the patients. ICART may be a breakthrough in the treatment of acute STEMI.

The intra-neuroendoscopic technique: a new method for rapid removal of acute severe intraventricular hematoma

The mortality rate of acute severe intraventricular hematoma is extremely high, and the rate of disability in survivors is high. Intraventricular hematoma has always been a difficult problem for clinical treatment. Although minimally invasive endoscopic hematoma evacuation is widely used to treat this disease, the technique still has room for improvement. Equipment for the intra-neuroendoscopic technique（INET） consists of two of our patented inventions： a transparent sheath（Patent No. ZL 200820046232.0） and a hematoma aspirator（Patent No. ZL 201520248717.8）. This study explored the safety and efficacy of INET by comparing it with extraventricular drainage in combination with urokinase thrombolytic therapy. This trial recruited 65 patients with severe intraventricular hemorrhage, including 35（19 men and 16 women, aged 53.2 ± 8.7 years） in the INET group and 30（17 men and 13 women, aged 51.5 ± 7.9 years） in the control group（extraventricular drainage plus urokinase thrombolytic therapy）. Our results showed that compared with the control group, the INET group exhibited lower intraventricular hemorrhage volumes, shorter intensive care-unit monitoring and ventricular drainage-tube placement times, and fewer incidences of intracranial infection, secondary bleeding, and mortality. Thus, the prognosis of survivors had improved remarkably. These findings indicate that INET is a safe and efficient new method for treating severe intraventricular hematoma. This trial was registered with Clinical Trials.gov（NCT02515903）.

The uPA/uPAR system in astrocytic wound healing

The repair of injured tissue is a highly complex process that involves cell prolife ration,differentiation,and migration.Cell migration requires the dismantling of intercellular contacts in the injured zone and their subsequent reconstitution in the wounded area.Urokinase-type plasminogen activator(u PA)is a serine proteinase found in multiple cell types including endothelial cells,smooth muscle cells,monocytes,and macrophages.A substantial body of experimental evidence with different cell types outside the central nervous system indicates that the binding of uPA to its receptor(uPAR)on the cell surface prompts cell migration by inducing plasmin-mediated degradation of the extracellular matrix.In contrast,although uPA and uPAR are abundantly found in astrocytes and u PA binding to uPAR triggers astrocytic activation,it is unknown if uPA also plays a role in astrocytic migration.Neuronal cadherin is a member of cell adhesion proteins pivotal for the formation of cell-cell conta cts between astrocytes.More specifically,while the extracellular domain of neuronal cadherin interacts with the extracellular domain of neuronal cadherin in neighboring cells,its intracellular domain binds toβ-catenin,which in turn links the complex to the actin cytos keleton.Glycogen synthase kinase 3βis a serine-threonine kinase that prevents the cytoplasmic accumulation ofβ-catenin by inducing its phosphorylation at Ser33,Ser37,and Ser41,thus activating a sequence of events that lead to its proteasomal degradation.The data discussed in this perspective indicate that astrocytes release u PA following a mechanical injury,and that binding of this u PA to uPAR on the cell membrane induces the detachment ofβ-catenin from the intracellular domain of neuronal cadherin by triggering its extracellular signal-regulated kinase 1/2-mediated phosphorylation at Tyr650.Remarkably,this is followed by the cytoplasmic accumulation ofβ-catenin because uPA-induced extracellular signalregulated kinase 1/2 activation also phosphorylates lipoprotein receptor-related protein 6 at Ser1490,which in turn,by recruiting glycogen synthase kinase 3βto its intracellular domain abrogates its effect onβ-catenin.The cytoplasmic accumulation ofβ-catenin is followed by its nuclear translocation,where it induces the expression of uPAR,which is required for the migration of astrocytes from the injured edge into the wounded area.

~1H-magnetic resonance spectroscopy screening for animals with acute cerebral infraction suitable forthrombolytic therapy

BACKGROUND： As a non-invasive technique which can provide comprehensive biological information, 1H-magnetic resonance spectroscopy （1H-MRS） may provide valuable reference data for irreversible recovery or reversible changes in ischemic tissue after stroke. OBJECTIVE： To monitor and evaluate the effect of the urokinase thrombolytic therapy after experimental acute cerebral ischemia by 1H-MRS technology and investigate its adaptability. DESIGN： Randomly controlled animal study. SETTINGS： Shenzhen Hospital of Peking University and National Key Laboratory of Pattern and Atom ＆ Molecular Physics, Wuhan Physics and Mathematics Institute, Chinese Academy of Science. MATERIALS： Eleven healthy adult Sprague-Dawley （SD） rats, weighing 260–300 g and of both genders, were supplied by Experimental Animal Center of Tongji Medical Collage, Huazhong University of Science and Technology [SCXK （e） 2004-007]. 4.7T superconducting nuclear magnetic resonance meter was provided by Brucker Company. METHODS： The experiment was carried out in Shenzhen Hospital of Peking University and National Key Laboratory of Pattern and Atom ＆ Molecular Physics, Wuhan Physics and Mathematics Institute, Chinese Academy of Science from August 2003 to December 2005. ① The rats were randomly divided into 30-minute self-thrombo-embolism group （n =6） and 60-minute self-thrombo-embolism group （n =5）. Six rats in 30-minute self-thrombo-embolism group were occluded with clot embolus for 30 minutes and 5 rats in 60-minute self-thrombo-embolism group were occluded for 60 minutes. 10 000 U/kg urokinase was dissolved in 2 mL saline and the operation lasted for 5 minutes. ② 1H-MRS was performed before thrombolysis and at 3 hours and 24 hours after successful embolization. The metabolic changes of N-acetyl-L-aspartic acid （NAA）/phosphocreatine （PCr） ＋ creatine （Cr）, choline phosphate （Cho）/PCr＋Cr and lactic acid （Lac）/PCr＋Cr in the region of interests were analyzed. ③ The T2W image was conducted 24 hours after the thrombolytic therapy with TR=500 ms and TE=25 ms. ④ The subjects were sacrificed immediately after 1H-MRS and the brain tissues were cut into pieces and stained with HE method; in addition, pathological changes were observed under optic microscope. MAIN OUTCOME MEASURES： ① Metabolic changes of NAA/PCr＋Cr, Cho/PCr＋Cr and Lac/PCr＋Cr in the region of interests; ② T2W image at 24 hours after the thrombolysis; ③ pathological observation of brain tissue. RESULTS： Eleven rats were all involved in the final analysis. ① Metabolic changes in the region of interests ： In 30-minute self-thrombo-embolism group, the Lac peak emerged immediately after the embolism, but the ischemic zone decreased 3 hours after the thrombolytic therapy （0.252±0.01, 0.603±0.01, P 〈 0.01）. Lac/（PCr＋Cr） ratio was 0.290±0.01 at 24 hours after thrombolysis, which was higher than that at 3 hours after thrombolysis （P 〈 0.01）. The NAA/ （PCr＋Cr） ratio decreased significantly at 3 hours after the thrombolysis as compared with that before thrombolysis （0.922±0.16, 1.196±0.01, P 〈 0.05）. In 60-minute self-thrombo-embolism group, the Lac/（PCr＋Cr） ratio was higher at 3 hours after thrombolysis than that before thrombolysis （0.846±0.12, 0.601±0.11, P 〈 0.05） and the NAA/（PCr＋Cr） decreased at 3 hours after the embolism. Fluctuation of NAA/ （PCr＋Cr） ranged from 0.68 to 0.75 before thrombolysis and from 0.71 to 0.75 at 3 hours after thrombolysis. ② T2W image： T2W image showed that 2 subjects in 30-minute self-thrombo-embolism group whose Lac/NAA was higher than 0.7 suffered from intracranial hemorrhage. This meant that the subjects with Lac/NAA 〉 0.7 were more likely to suffer from intracranial hemorrhage. ③ Histological and morphological examinations： Optic microscope demonstrated that interspace surrounding nerve cells was widened at ischemic center; neurons were swelling; nucleus was stained lightly; pyknosis and mesenchymal edema were mainly observed in lateral cortex of brow and vertex and in lateralpart of corpus striatum. CONCLUSION： ①Compound parameters in ischemic area before thrombolysis should be regarded as an important predicting marker for thrombolytic therapy, effect evaluation and termination. ② 1H-MRS combining with other imaging technique is a detecting way for screening cases who are suitable for thrombolytic therapy.

Cloning and Analysis of the Promoter Region of Rat uPA Gene

Objective To clone and analyze the promoter sequence of rat urokinase plasminogen activator protein gene. Methods The genomic DNA was extracted from rat testicular tissue. According to urokinase plasminogen activator, the gene sense primer and antisense primer of uPA gene were designed and synthesized, then Touch-Down PCR were performed. After proper purification, the PCR product was sequenced, analyzed with the promoter prediction software and compared with the DNA sequence of rattuas urokinase plasminogen activator. Results The cloned uPA gene was about 1 572 bp in length, which contained a full open-reading frame with 21 bp in length exons, and the upper region of transcriptional start was 1 551 bp in length which was eucaryon transcriptional control area. The 5＇ UTR had a promoter region including a non-responsive TATA-box. Not only the GC-box binding region was found in this gene, but also active protein I （AP1） and SP1 were seen in other regions. Conclusion A 1 572 bp uPA gene fragment （GenBank accession No.X65651） was obtained from rat genomic DNA library, containing eucaryon transcriptional control area with a promoter region, non-conspicuous TATA-box, GC-box and an extron. A non-responsive TATA-box is located at the upper -30 region.