• Title/Summary/Keyword: Urokinase

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Relationship between Plasminogen Activity and Plasminogen Inhibitor during the Culture of Porcine Oviduct Epithelial Cells

  • Ahn, Shin-Hye;Cheong, Hee-Tae;Yang, Boo-Keun;Kim, Dae-Young;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.33 no.4
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    • pp.203-209
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    • 2009
  • The present study was performed to identify changes of plasminogen activator (PA) and plasminogen activator inhibitor (PAI) in porcine oviduct epithelial cells (POECs) during the estrous cycle. POECs obtained from ovary in pre-ovulatory (Pre-Ov), early to mid-luteal stage (Early-mid L) and post-ovulatory stage (Post-Ov). For the examine of PA activity, $1{\times}10^5$ fresh cells of POECs were cultured in DMEM/Ham F-12 containing 10% FBS and 0.2% amphotericin under humidified atmosphere of 5% $CO_2$ in air and $38^{\circ}C$. The urokinase-type PA (uPA) was observed at 7 days of POECs culture. PA activity was measured with culture prolonged of 0, 3, 6, 12 and 24 h after culture of 7 days. The PA activity were high significantly (p<0.05) at 12 h of culture, but PA activity were decreased with culture periods increased. The PA activity in POECs of Post-Ov stage were higher significantly (p<0.05) than that of Early-mid L and Pre-Ov stage. When PAI-1 and PAI-2 were added during the POECs culture, the PA were observed significant low activity (p<0.05). The PA activity and protein expression were decreased by PA inhibitor. This results suggest that PAI-1 and PAI-2 have a suppressive action on change of PA activity during the estrous cycle of pigs. Specifically, this study using PA inhibitor was effect the PA activity and PAI expression in oviduct epithelial cells in pigs.

The Development of Serum Free Medium for the Economic Production of scu-PA from HEK Cells (인간 신장세포로부터 scu-PA의 경제적 생산을 위한 무혈청 배지의 개발)

  • 김현규;김현구
    • KSBB Journal
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    • v.9 no.5
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    • pp.518-524
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    • 1994
  • Serum free medium was developed to cultivate Human Embryonic Kidney cells with Cytodex III microcarriers. The cells normally attached and spreaded on the microcarriers in serum free medium, and grew well by bead to bead processes. 85% of attachment yield was obtained on Cytodex III in this medium, compared to about 93% in 1% serum containing medium. About 90% of the attached HEK cells spreaded after 6 hours of post-attachment periods on the surface of microcarrier. Maximum cell density and scu-PA concentration in a serum free medium were $9.1{\times}10^5$ cells/ml and 1790 IU/ml, respectively, with fed-batch cultivation. Maximum cell density and scu-PA concentration in this medium with perfusion cultivation were $2.5{\times}10^6$ cells/ml and 1820 IU/ml, respectively. The conversion of single chain urokinase type plasminogen activator (scu-PA) into two chain type plasminogen activator (tc-PA) was less than 5% in a serum free medium compared to about 10% in 0.5% serum containing medium.

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Effect of Progesterone on Expression of Prostaglandin Synthases and Plasminogen Activator in Bovine Endometrium during Estrous Cycle (발정주기의 소 자궁내막에서 Progesterone이 Prostaglandin 합성효소와 Plasminogen Activator 발현에 미치는 영향)

  • Choi, Su-Bin;Hwangbo, Yong;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Journal of Embryo Transfer
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    • v.31 no.1
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    • pp.53-59
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    • 2016
  • This study was to investigate effect of progesterone ($P_4$) on prostaglandin (PG) synthases and plasminogen activators (PAs) system in bovine endometrium during estrous cycle. Endometrium tissues were collected from bovine uterus on follicular and luteal phase and were incubated with culture medium containing 0 (Control), 0.2, 2, 20 and 200 ng/ml $P_4$ for 24 h. The $PGF_{2{\alpha}}$ synthase (PGFS), $PGE_2$ synthase (PGES), cyclooxygenase-2 (COX-2), urokinase PA (uPA), and PA inhibitors 1 (PAI-1) mRNA in bovine endometrium were analyzed using reverse transcription PCR and PA activity was measured using spectrophotometry. In results, COX-2 was higher at 2 ng/ml $P_4$ group than control group in luteal phase (p<0.05), but, it did not change in follicular phase. Contrastively, PGES was significantly increased in 2 ng/ml $P_4$ group compared to control group in follicular phase, but there were no significant differ among the treatments in luteal phase. uPA was no significant difference between $P_4$ treatment groups and control group in both of different phase. PAI-1 was decreased in 20 ng/ml $P_4$ group compared to control group in follicular phase (p<0.05). PA activity was decreased in 2 ng/ml $P_4$ group compared to other groups in follicular and luteal phase (p<0.05). In conclusion, we suggest that $P_4$ may influence to translation and post-translation process of PG production and PA activation in bovine endometrium.

Antiangiogenic and Antitumor Activities of the Cryptic Fragments with Kringle Architecture

  • Joe, Young-Ae;Kim, Myung-Rae;Shim, Byoung-Shik;Oh, Dae-Shik;Hong, Sung-Hee;Hong, Yong-Kil
    • Biomolecules & Therapeutics
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    • v.11 no.4
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    • pp.205-213
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    • 2003
  • Various angiogenesis inhibitors target vascular endothelial cells and block tumor angiogenesis. Angiostatin is a specific endogenous angiogenesis inhibitor in clinical trials, which contains only the first four triple loop structures, known as kringle domains. Its generated by proteolytic cleavage of its parent molecule plasminogen, which itself does not exhibit antiangiogenic activity. Kringle domains from prothrombin, apolipoprotein, hepatocyte growth factor, urokinase and tissue-type plasminogen activator also elicit anti-angiogenic or antitumor activities in several model systems, albeit low amino acid sequence identity between angiostatin and each individual kringle. However, the differential effects of each kringle domain on endothelial cell proliferation, and migration observed in these kringle domains, suggest that the amino acid sequence of the primary structure is still important although kringle architecture is essential for anti-mlgiogenic activity. If it is further studied as to how amino acid sequence and kringle architecture contributes in anti-angiogenic activity, with studies on underlying mechanisms of anti-angiogenesis by kringle-based angiogenesis inhibitors, it will provide basis for the development of new potent anti-angiogenesis inhibitors and improvement of the efficacy of angiogenesis inhibitors.

Clinical Analysis of Stereotactic Aspiration and Conservative Management in Spontaneous Thalamic Hematoma (자발성 시상부 출혈에서 뇌정위적 흡인술 및 보존적 치료의 임상분석)

  • Nam, Cheon Hyun;Kang, Jae Kyu;Doh, Jong Oung;Lee, Chun Dae
    • Journal of Korean Neurosurgical Society
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    • v.30 no.2
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    • pp.156-162
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    • 2001
  • Objective : The purpose of this study is to review our experience with spontaneous thalamic hemorrhage. Clinical outcome of patients was brought about by comparing stereotactic aspiration and conservative medical therapy. Methods : The study consists of seventy-three cases with spontaneous thalamic hemorrhage which were treated from the period of Jan. 1993 to Dec. 1999. Thirty-eighty patients were treated with computed tomography (CT) guided stereotactic aspiration and thirty-five patients were treated conservatively. We compared the factors affecting treatment and the factors are as follows : age & sex, conscious level on admission, hematoma volume, hematoma sites, presence of ventricular penetration. Results : The results in the thirty eight stereotactic aspirated cases for the 6 months from oneset are as follows : good recovery or moderate disability in 43%, severe disability in 32%, vegitative state in 11%, dead in 13% respectively. The clinical result was more favorable in stereotactic aspiration, with 11-30cc hematoma volume, extend to internal capsule of hematoma, poor conscious level on admission than conservative medical therapy. But age & sex, conscious level on admission, presence of ventricular penetration were not influential in the statistical outcome between stereotactic aspiration and conservative medical therapy. Conclusion : Treatment modality of spontaneous thalamic hemorrhage is still controversial. But stereotactic aspiration is more recommended for improvement therapeutic results than conservative treatment or open craniotomy in case of 11-30cc hematoma volume, extend to internal capsule of hematoma and poor conscious level on admission.

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Effects of Progesterone and 17β-Estradiol under Presence or Absence of FBS on Plasminogen Activators Activity in Porcine Uterine Epithelial Cells

  • Hwangbo, Yong;Lee, Mi-Rim;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Development and Reproduction
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    • v.22 no.4
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    • pp.309-318
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    • 2018
  • The present study was conducted to investigate the regulatory mechanism of plasminogen activators (PAs) activation by $17{\beta}$-estradiol ($E_2$) and progesterone ($P_4$) in porcine uterine epithelial cells (pUECs). pUECs were collected from porcine uterine horn and cultured at 80% confluence. Then, 0.1% (v/v) DMSO, 20 ng/mL $E_2$, and $P_4$ with or without fetal bovine serum (FBS) treated to cultured cells for 24 hours. The supernatants were used for measurement of PAs activity and expression of urokinase-type PA (uPA), tissue-type PA (tPA), uPA specific receptor (uPAR), and type-1 PA inhibitor (PAI-1) mRNA were analyzed by real-time PCR. The expression of PAs-related genes was not affect by steroid hormones in both of serum treatment groups. However, PAs activity was increased by treatment of $E_2$ compared to 0.1% DMSO treatment in serum-free group (p<0.05). Then, $E_2$ and $P_4$ were diluted with 0.002% (v/v) DMSO for reduction of its effect and treated to cultured cells without FBS. Only tPA mRNA was significantly increased by $E_2$ treatment (p<0.05). PAs activity was enhanced in $E_2$ treated group compared to control groups (p<0.05). These results indicate that serum-free condition is more proper to evaluate effect of steroid hormones and activation of PAs in pUECs was mainly regulated by estrogen. These regulation of PAs activation may be associated with uterine remodeling during pre-ovulatory phase in pigs, however, further studies are needed to investigate precise regulatory mechanism.

The Effect of Locally Administered Fibrinolytic Drugs Following Aneurysmal Subarachnoid Hemorrhage : A Meta-Analysis with Eight Randomized Controlled Studies

  • Jang, Kyoung Min;Choi, Hyun Ho;Nam, Taek Kyun;Park, Yong Sook;Kwon, Jeong Taik;Byun, Jun Soo;Hwang, Doyeon
    • Journal of Korean Neurosurgical Society
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    • v.64 no.2
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    • pp.207-216
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    • 2021
  • Objective : Rapid dissolution of blood clots reduces vasospasm and hydrocephalus after subarachnoid hemorrhage (SAH), and locally administered fibrinolytic drugs (LAFDs) could facilitate the dissolution. However, the efficacy of LAFDs remains controversial. The aim of this meta-analysis was to determine the efficacy of LAFDs for vasospasm and hydrocephalus and in clinical outcomes. Methods : From PubMed, EMBASE, and Cochrane database, data were extracted by two authors. Meta-analysis was performed using a random effect model. Inclusion criteria were patients who had LAFDs with urokinase-type or recombinant tissue-plasminogen activator after SAH in comparison with medically untreated patients with fibrinolytic drugs. We only included randomized controlled trials (RCTs) in this analysis. The outcomes of interest were vasospasm, hydrocephalus, mortality, and 90-day unfavorable functional outcome. Results : Data from eight RCTs with 550 patients were included. Pooled-analysis revealed that the LAFDs were significantly associated with lower rates of vasospasm (LAFDs group vs. control group, 26.5% vs. 39.2%; odds ratio [OR], 0.48; 95% confidence interval [CI], 0.32-0.73); hydrocephalus (LAFDs group vs. control group, 26.0% vs. 31.6%; OR, 0.54; 95% CI, 0.32-0.91); and mortality (LAFDs group vs. control group, 10.5% vs. 15.7%; OR, 0.58; 95% CI, 0.34-0.99). The proportion of 90-day unfavorable outcomes was lower in the LAFDs group (LAFDs group vs. control group, 32.7% vs. 43.5%; OR, 0.55; 95% CI, 0.37-0.80). Conclusion : This meta-analysis with eight RCTs indicated that LAFDs were significantly associated with lower rates of vasospasm and hydrocephalus after SAH. Thus, LAFDs could consequently reduce mortality and improve clinical outcome after SAH.

mRNA Expression Differences of uPA, uPAR in Eutopic Endometrium of Advanced Stage Endometriosis Patients (중증 자궁내막증 환자의 자궁내막과 정상인 자궁내막에서 uPA, uPAR mRNA 발현의 차이에 관한 연구)

  • Hur, Sung Eun;Lee, Ji Young;Lee, Woon Jung;Chung, Hye-Sung;Chung, Hye Won
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.4
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    • pp.229-236
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    • 2006
  • Objective: We investigated the expression of uPA and uPAR in eutopic endometrium of advanced stage endometriosis and control patients. Methods: The 33 endometriosis patients and 32 controls were enrolled. Endometrial samples were obtained from 65 premenopausal women aged 29-44 years, undergoing laparoscopic surgery or hysterectomy for non-malignant lesions. Sufficient samples were collected from 33 patients with endometriosis stage Ill and IV and 32 controls without endometriosis confirmed by laparoscopic surgery. The mRNA expression of uPA and uPAR from eutopic endometrium were analyzed by RT-QC PCR. Results: The mRNAs of uPA and uPAR were expressed in eutopic endometrium from endometriosis and normal controls throughout the menstrual cycle. Uterine endometrium from women with endometriosis expresses significantly (p<0.05) higher levels of u-PA mRNA than endometrium from normal women without endometriosis in the proliferative phase. There were no significant differences in expression of uPAR in eutopic endometrium between controls and endometriosis patients. Conclusion: These results suggest that eutopic endometrium from endometriosis patients may be more invasive and prone to peritoneal implantation because of greater u-PA mRNA expression than endometrium from women without endometriosis. Thus, increased proteolytic activity may be one etiology for the invasive properties of the endometrium resulting in the development of endometriosis.

Pharmaco-mechanical Thrombectomy and Stent Placement in Patients with May-Thurner Syndrome and Lower Extremity Deep Venous Thrombosis (May-Thurner 증후군과 동반된 하지 심부정맥혈전환자에서 혈전제거술과 스텐트삽입술)

  • Jeon, Yonh-Sun;Kim, Yong-Sam;Cho, Jung-Soo;Yoon, Yong-Han;Baek, Wan-Ki;Kim, Kwang-Ho;Kim, Joung-Taek
    • Journal of Chest Surgery
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    • v.42 no.6
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    • pp.757-762
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    • 2009
  • Background: Compression of the left common iliac vein by the overriding common iliac artery is frequently combined with acute deep vein thrombosis in patients with May-Thurner Syndrome. We evaluate the results of treatment with thrombolysis and thrombectomy followed by stenting in 34 patients with May-Thurner Syndrome combined with lower extremity deep venous thrombosis. Material and Method: The authors retrospectively reviewed the records of 34 patients (mean age: $65{\pm}14$ year old) who had undergone stent insertion for acute deep vein thrombosis that was caused by May-Thurner syndrome. After thrombectomy and thrombolysis, insertion of a wall stent and balloon angioplasty were performed to relieve the compression of the left common iliac vein. Urokinase at a rate of 80,000 to 120,000 U/hour was infused into the thrombosed vein via a multi-side hole thrombolysis catheter. A retrieval inferior vena cava (IVC) filter was placed to protect against pulmonary embolism in 30 patients (88%). Oral anticoagulation with warfarin was maintained for 3 months, and follow-up Multi Detector Computerized Tomography (MDCT) angiography was done at the date of the patients' hospital discharge and at the 6 months follow-up. Result: The symptoms of deep venous thrombosis disappeared in two patients (4%), and there was clinical improvement within 48 hours in twenty eight patients (82%), but there was no improvement in four patients (8%). The MDCT angiography at discharge showed no thrombus in 9 patients (26%) and partial thrombus in 21 (62%), whereas the follow-up MDCT at $6.4{\pm}5.5$ months (32 patients) revealed no thrombus in 23 patients (72%), and partial thrombus in 9 patients (26%). Two patients (6%) had recurrence of DVT, so they underwent retreatment. Conclusion: Stent insertion with catheter-directed thrombolysis and thrombectomy is an effective treatment for May-Thurner syndrome combined with acute deep vein thrombosis in the lower extremity.

Radiation Induced Changes in the Expression of Fibronectin, Pai-1, MMP in Rat Glomerular Epithelial Cell (백서 사구체 상피세포에서 방사선에 의한 Fibronectin, Pai-1, MMP 발현의 변화)

  • Park Woo-Yoon;Kim Won-Dong;Zheng Ying;Ha Tae-Sun;Kim Jae-Sung;Cho Moon-June
    • Radiation Oncology Journal
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    • v.24 no.1
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    • pp.58-66
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    • 2006
  • Purpose: Renal irradiation can lead to the development of radiation nephropathy, and this is characterized by the accumulation of extracellular matrix and final fibrosis. To determine the possible role of the glomerular epithelial cell, the radiation-induced changes in the expression of its genes associated with the extracellular matrix were analyzed. Materials and Methods: Rat glomerular epithelial cells (GEpC) were irradiated with a single dose of 0, 2, 5, 10 and 20 Gy with using 6 MV LINAC (Siemens, USA), and the samples were collected 6, 24, 48 and 72 hours post-irradiation, respectively. Northern blotting, western blotting and zymography were used to measure the expression level of fibronectin (Fn), plasminogen activator inhibitor-1 (Pai-1), matrix metalloproteinases-2, 9 (MMP-2, 9), tissue inhibitor of metalloproteinase-2 (TIMP-2), tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator (u-PA). Results: Irradiation with a single dose of 10 Gy resulted in a significant increase in Fn mRNA since 24 hours post-irradiation, and a single dose of 5 and 10 Gy significantly increased the Fn immunoreactive protein measured 48 hours post-irradiation. An increase in Pai-1 mRNA and protein was also observed and especially, a single dose of 10 Gy significantly increased the mRNA measured 24 and 48 hours post-irradiation. The active MMP-2 measured 24 hours post-irradiation slightly increased in a dose dependent manner, but this increase did not reach statistical significance. The levels of MMP-9, TIMP-2, t-PA and u-PA appeared unaltered after irradiation. Conclusion: Irradiation of the glomerular epithelial cells altered the expression of genes associated with the extracellular matrix, implying that the glomerular epithelial cell may be involved in the development of radiation nephropathy.