• Journal of Korean Neurosurgical Society
• /
• v.37 no.4
• /
• pp.287-292
• /
• 2005

Objective: Recent clinical studies have demonstrated that intracisternal administration of recombinant tissue plasminogen activator(rt-PA) can facilitate the normal clearing of blood from the subarachnoid space. Urokinase, a first generation fibrinolytic agent, has been used to liquify such clots with some success. Therefore, recombinant tissue plasminogen activator, a second generation fibrinolytic drug that may be safer and more effective, is studied to evaluate its dosage to lyse clots in vitro and reactivity in the brain parenchyme. Methods: Intracerebral hematomas were created by stereotactically injecting 2ml of clotted autogenous blood into the brain parenchyme of total 28 anesthetized adult cats (weighting 3.8 to 4.1 kg). The control animals (group A) received 1 ml of normal saline injected into the clots and the experimental animals received each 0.1 mg of rt-PA (group B), 0.5mg of rt-PA (group C) and 1 mg of rt-PA (group D) at 6 hours after the clot injection. Results: 1. The amount of remained clots after lysing the hematomas were as follows: $1.80{\pm}0.17ml$ in group A, $1.65{\pm}0.23ml$ in group B, $0.61{\pm}0.37ml$ in group C and $0.52{\pm}0.34$ in group D. The result indicated that hematomas in rt-PA treated groups (C & D) were lysed better than the control group. 2. At least 0.5mg of rt-PA should be required for the lysis of 2ml of hematomas. 3. Light microscopic examination revealed no histological evidence of hemorrhage in tissue sections from each brain. Conclusion: Recombinant tissue plasminogen activator may be safely and effectively employed for the lysis of intracerebral hematomas in animal model.

• Journal of Veterinary Clinics
• /
• v.33 no.3
• /
• pp.155-159
• /
• 2016

Four dogs were brought to the Veterinary Medicine Teaching Hospital of Seoul National University (VMTH SNU) with a history of hind limb ataxia, three with pain, one without pain. Three of the four showed weak to absent femoral pulses and cold extremities. Thromboembolism was identified by ultrasonography in the external and/or internal iliac arteries. A thrombolytic agent, recombinant tissue plasminogen activator (rt-PA), was administered (0.5-1 mg/kg, every 60-120 min, 3-5 doses). Two dogs (Cases 2 and 3), which were instantly provided rt-PA treatment, survived 6 and 17 months, respectively, although hematemesis and hematochezia were observed during treatment. In the other two dogs (Cases 1 and 4), rt-PA was administered 4 and 28 days after the appearance of pelvic limb symptoms, which may have limited the benefits of the treatment. When rt-PA treatment is instituted instantly and the side effects are monitored thoroughly during treatment, a good prognosis might be expected in canine aortic thromboembolism. For this reason, we suggest that rt-PA treatment should be initiated immediately if thromboembolism is identified.

• Development and Reproduction
• /
• v.3 no.1
• /
• pp.29-38
• /
• 1999

The pathogenesis of endometriosis is unknown, but retrograde menstruation is widely accepted as an etiology. Refluxed endometrium from endometriosis patients is more prone to implant and invade peritoneum possibly through the action of extracellular proteolysis. This proteolytic action may involve plasminogen activators and the collagenase system. Plasminogen activators (PAs) and matrix metalloproteinases (MMPs) play a critical role in the breakdown of extracellular matrix components and basement membrane in the processes of implantation and tumor invasion. PAs are inhibited by plasminogen activator inhibitor (PAI) and MMPs activity is inhibited by tissue inhibitor of metalloproteinase (TIMP). To test the hypothesis that lower expression of PAI-1 and TIMP-3 in endometrium from women with endometriosis, we investigated their PAI-1 and TIMP-3 expression by quantitative competitive RT PCR in endometrium from women with and without endometriosis. Endometrial tissues were obtained from 14 patients with severe endometriosis and 14 patients without endometriosis. Total RNA was extracted and reverse transcribed into cDNA, and quantitative competitive PCR (QC PCR) was performed to evaluate PAI-1 and TIMP-3 mRNA expression. Endometrium from patients with endometriosis showed decreased expression of PAI-1 and TIMP-3 mRNA compared to endometrium from control in luteal phase (p<0.05). Our results suggest that endometrium from women with endometriosis expresses lower levels of PAI-1 and TIMP-3 than endometrium from normal women. Endometrium from endometriosis patients may be more invasive and prone to peritoneal implantation than control because of higher PA and MMP enzymatic activity. Thus, increased proteolytic activity may be one of the reasons for the invasive properties of the endometrium resulting in the development of endometriosis.

• Clinical and Experimental Reproductive Medicine
• /
• v.33 no.4
• /
• pp.229-236
• /
• 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.

• Journal of Plant Biotechnology
• /
• v.36 no.1
• /
• pp.30-37
• /
• 2009

Tissue-type plasminogen activator (t-PA) is a thrombolytic agent important in fibirn clot lysis. T-PA causes fibirn-specific plasminogen activation. Six binary vectors harboring t-PA and its derivative genes were cloned and expressed in transgenic alfalfa plants. The insertion of the t-PA and its derivative genes in genomic DNA of alfalfa plants was confirmed by PCR. The presence of the t-PA and its derivative transcripts in total RNAs of the transgenic alfalfa leaves was verified by RT-PCR. ELISA experiments demonstrated that the highest level of recombinant t-PA expression was $75.1{\mu}g$/ total soluble protein (mg) in alfalfa plants. The amount of recombinant t-PA and its derivative proteins in transgenic plants was estimated to range from 9.7 to $39.5{\mu}g$/ total soluble proteins (mg). Western blot analysis of the transformed alfalfa leaves revealed bands of approximately 68-kDa recombinant t-PA and its derivative proteins. The fibrinolysis of recombinant t-PA and its derivative proteins was confirmed by a fibrin plate assay (range from 3.2 to 8.1 cm). The results presented provide information for the development of an additional production of recombinant human proteins having pharmaceutical applications using transgenic plants.

• Journal of Gastric Cancer
• /
• v.4 no.4
• /
• pp.207-212
• /
• 2004

Purpose: Invasion and metastasis in solid tumors require the action of tumor-associated proteases. The serine protease urokinase-type plasminogen (uPA) and receptor (uPAR) appear to have a major function in these processes. Expression of the uPAR is elevated in breast and colon carcinomas, and this is often associated with invasiveness and poor prognosis. The purpose of this study was to determine whether the expression of the uPAR gene correlates with clinico-pathological parameters in human gastric carcinomas. Materials and Methods: We examined the expression of uPAR mRNA by using northern blot analysis and RT-PCR in 35 gastric carcinomas and the surrounding normal mucosa. Macroscopic and histopathological tumor findings and survival rates were obtained from the patient records and from endoscopic, surgical, and pathological reports. Results: The expression of uPAR and was higher in most neoplasms than in the corresponding normal mucosal tissue. uPAR mRNA expression in tumors correlated well with lymph-node metastasis (P<0.02) and tumor stage (P<0.01). The survival rate of patients with tumors displaying high uPAR expression levels was significantly lower (P<0.04) than that of patients without uPAR expression, but IL-8 showed only the tendency of survival difference. Conclusion: These results suggest that uPAR may be an important prognostic factor in human gastric carcinomas.

• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.9
• /
• pp.5403-5408
• /
• 2013

Objective: CXCL12 exerts a wide variety of chemotactic effects on cells. Evidence indicates that CXCL12, in conjunction with its receptor, CXCR4, promotes invasion and metastasis of tumor cells. Our objective was to explore whether the CXCL12-CXCR4 biological axis might influence biological behavior of pancreatic cancer cells. Methods: Miapaca-2 human pancreatic cancer cells were cultured under three different conditions: normal medium (control), medium + recombinant CXCL12 (CXCL12 group), or medium + CXCR4-inhibitor AMD3100 (AMD3100 group). RT-PCR was applied to detect mRNA expression levels of CXCL12, CXCR4, matrix metalloproteinase 2 (MMP-2), MMP-9, and human urokinase plasminogen activator (uPA). Additionally, cell proliferation and invasion were performed using CCK-8 colorimetry and transwell invasion assays, respectively. Results: CXCL12 was not expressed in Miapaca-2 cells, but CXCR4 was detected, indicating that these cells are capable of receiving signals from CXCL12. Expression of extracellular matrix-degrading enzymes MMP-2, MMP-9, and uPA was upregulated in cells exposed to exogenous CXCL12 (P<0.05). Additionally, both proliferation and invasion of pancreatic cancer cells were enhanced in the presence of exogenous CXCL12, but AMD3100 intervention effectively inhibited these processes (P<0.05). Conclusions: The CXCL12-CXCR4 biological axis plays an important role in promoting proliferation and invasion of pancreatic cancer cells.