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Neuroprotective Effect of Dizocilpine (MK-801) via Anti-apoptosis on Hypoxic-ischemic Brain Injury in Neonatal Rats

  • Seo, Min-Ae;Lee, Hyun-Ju;Choi, Eun-Jin;Kim, Jin-Kyung;Chung, Hai-Lee;Kim, Woo-Taek
    • Neonatal Medicine
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    • v.17 no.2
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    • pp.181-192
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    • 2010
  • Purpose: Current studies have demonstrated the neuroprotective effects of dizocilpine (MK-801) in many animal models of brain injury, including hypoxic-ischemic (HI) encephlopathy, trauma and excitotoxicity, but limited data are available for those during the neonatal periods. Here we investigated whether dizocilpine can protect the developing rat brain from HI injury via anti-apoptosis. Methods: In an in vitro model, embryonic cortical neuronal cell culture of Sprague-Dawley (SD) rats at 18-day gestation was done. The cultured cells were divided into three groups: normoxia (N), hypoxia (H), and hypoxia treated with dizocilpine (HD). The N group was prepared in 5% $CO_2$ incubators and the other groups were placed in 1% $O_2$ incubators (94% N2, 5% $CO_2$) for 16 hours. In an in vivo model, left carotid artery ligation was done in 7-day-old SD rat pups. The animals were divided into six groups; hypoxia (N), hypoxia (H), hypoxia with sham-operation (HS), hypoxia with operation (HO), HO treated with vehicle (HV), and HO treated with dizocilpine (HD). Hypoxia was made by exposure to a 2 hour period of hypoxic incubator (92% N2, 8% $O_2$). Results: In the in vitvo and in vivo models, the expressions of Bcl-2 in the hypoxia groups were reduced compared to the normoxia group. whereas those in the dizocilpine-treated group were increased compared to the hypoxia group. However. the expressions of Bax and caspase-3 and the ratio of Bax/Bcl-2 were revealed reversely. Conclusion: Dizocilpine has neuroprotective property over perinatal HI brain injury via anti-apoptosis.

Cloning of the β-Lactamase Gene from Bacillus sp. J105 and Analysis of Its Expression in E. colis Cells (Bacillus sp. J105 유래 β-lactamase 유전자의 cloning 및 E. coli 내에서의 발현 분석)

  • Kang, Won-Dae;Lim, Hak-Seo;Seo, Min-Jeong;Kim, Min-Jeong;Lee, Hye-Hyeon;Cho, Kyeong-Soon;Kang, Byoung-Won;Seo, Kwon-Il;Choi, Yung-Hyun;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.18 no.11
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    • pp.1592-1599
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    • 2008
  • The $\beta$-lactamase gene was cloned into E. coli DH5$\alpha$ from Bacillus sp. J105 with strong resistance against $\beta$-lactam antibiotics. The chromosomal DNA was partially digested with Sau3AI and ligated to BamHI digested pLAFR3. $\beta$-Lactamase positive clones were obtained by using in vitro packaging kit. The pKL11-${\Delta}4.6$ with $\beta$-lactamase activity was obtained by subcloning of the recombinant plasmid ($\beta$-lac +). The 6.5 kb fragment in the subcloned plasmid was sequenced. The DNA fragment that contains the $\beta$-lactamase gene encodes 309 amino acids. The 0.17 kb upstream region was similar to those of B. thuringinesis and B. cereus with 97% identity. The deduced amino acids sequence was also similar to those of $\beta$-lactamase from B. thuringinesis and B. cereus with 97% and 94% identity, respectively. The phylogenetic tree also showed the relationships of the $\beta$-lactamase gene of Bacillus sp. J105 to genetically related that of other Bacillus strains. Analysis of expression pattern of the pKL11-${\Delta}4.6$ in E. coli, revealed that the secretion efficiency of $\beta$-lactamase was $4{\sim}5%$ and the molecular weight was as same as that of original $\beta$-lactamase (31 kDa) from Bacillus sp. J105.

Clinical Application of Stent-graft in Thoracic Aortic Diseases (흉부 대동맥 질환에서 스텐트-그라프트의 임상적 적용)

  • Kim, Kyung-Hwan;Lee, Cheul;Chang, Ji-Min;Chung, Jin-Wook;Ahn, Hyuk;Park, Jae-Hyung
    • Journal of Chest Surgery
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    • v.34 no.9
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    • pp.698-703
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    • 2001
  • Background: Endovascular stent-graft insertion in aortic diseases is now generally accepted as an attractive alternative treatment modality. We reviewed our clinical experiences of endovascular stent-graft insertion in thoracic aorta. Material and Method: Since 1995, we performed 8 cases of endovasclar stent-graft insertion. Preoperative diagnoses were aortic aneurysms in 4, traumatic aortic ruptures in 3, and ruptured aortic pseudoaneurysm in 1. All procedures were performed in angiography room with the guidance of fluoroscopy. The stent-graft device is a custom-made 0.35mm thickness Z-shaped stainless steel wires, intertwined with each other using polypropylene suture ligation. It is covered with expanded Dacron vascular graft. Result: All procedures were performed successfully. Follow-up studies revealed 2 minimal perigraft leakages. There was no significant leakage or graft migration. 2 patients expired due to multiple organ failure and fungal sepsis. Other survivors(6) are doing well. Conclusion: Endovascular stent-graft insertion is relatively saft and effective treatment modality in the managment of various types of aortic diseases. In may be an effective alternative in aortic diseases of great surgical risk.

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The Validity of a Combined High Saphenous Division and Sclerotherapy for Varicose Vein (하지정맥류의 치료에 있어서 복재정맥 분리 결찰 및 혈관 경화요법 병용의 유용성)

  • Choi Se-Yong;Yang In-Suk;Won Tae-Hee
    • Journal of Chest Surgery
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    • v.39 no.7 s.264
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    • pp.544-548
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    • 2006
  • Background: The purpose of this study is to evaluate the validity of a combined high saphenous vein division and sclerotherapy without sacrificing saphenous vein for varicose vein. Material and Method: Between August 2004 and October 2005, 70 limbs in 50 patients were treated by a combined high saphenous vein division and sclero-therapy. The operative indication is valvular incompetence of femoral-saphenous or popliteal-saphenous junction. Patients received local anesthesia and were treated by a combined high saphenous vein division, ligation of incompetent perforating vein and $1{\sim}3%$ sclerosant. Patients received 1 day hospitalization and applied in com-pression stocking for 6 weeks. Patients followed after 1 week, 1 and 3 months. Result: Mean age of patients was $50{\pm}11$. The female was more common. 8 patients was no symptom, another 42 patients complained of pain, heaviness and fatigue of limbs. The symptoms of varicose vein disappeared 1 month after the procedure in all symptomatic patients. 8 patients needed a adjuvant sclerotherapy for residual varicose vein on 1 week after the procedure. There were only minor complications such as hematoma (1), wound infection (1), thrombophlebitis (20), skin blister (10), hyper-pigmentation (1), and skin ulcer (1). Conclusion: We concluded that a combined high saphenous vein division and sclerotherapy without sacrificing saphenous vein for varicose vein is simple, less invasive, economical, and effective treatment for primary varicose vein, and it has a special advantage that saphenous vein can be used as a bypass conduit later. The method was selective in old aged patients.

The Neuroprotective Effects of 6-cyano-7-nitroquinoxalin-2,3-dione (CNQX) Via Mediation of Nitric Oxide Synthase on Hypoxic-ischemic Brain Injury in Neonatal Rats

  • Jung, Ji-Eun;Keum, Kyung-Hae;Choi, Eun-Jin;Kim, Jin-Kyung;Chung, Hai-Lee;Kim, Woo-Taek
    • Neonatal Medicine
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    • v.18 no.1
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    • pp.59-69
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    • 2011
  • Purpose: Current studies have demonstrated the neuroprotective effects of 6-cyano-7-nitroquinoxalin-2,3-dione (CNQX) in many animal models of brain injury, including hypoxic-ischemic (HI) encephlopathy, trauma and excitotoxicity, but limited data are available for those during the neonatal periods. Here we investigated whether CNQX can protect the developing rat brain from HI injury via mediation of nitric oxide synthase. Methods: In an in vivo model, left carotid artery ligation was done in 7-day-old Sprague-Dawley (SD) rat pups. The animals were divided into six groups; normoxia (N), hypoxia (H), hypoxia with sham-operation (HS), hypoxia with operation (HO), HO treated with vehicle (HV), and HO treated with CNQX at a dose of 10 mg/kg (HC). Hypoxia was made by exposure to a 2 hr period in the hypoxic chamber (92% $N_2$, 8% $O_2$). In an in vitro model, embryonic cortical neuronal cell culture of SD rats at 18-day gestation was done. The cultured cells were divided into three groups: normoxia (N), hypoxia (H), and hypoxia treated with CNQX (HC). The N group was prepared in 5% $CO_2$ incubators and the other groups were placed in 1% $O_2$) incubators (94% $N_2$, 5% $CO_2$) for 16 hr. Results: In the in vitvo and in vivo models, the expressions of iNOS and eNOS were reduced in the hypoxia group when compared to the normoxia group, whereas they were increased in the CNQX-treated group compared to the hypoxia group. In contrast, the expression of nNOS was showed reversely. Conclusion: CNQX has neuroprotective property over perinatal HI brain injury via mediation of nitric oxide synthase.

The Effect of Postnatal Dexamethasone Treatment on Hypoxic-Ischemic Brain Injury in Neonatal Rats (신생쥐의 저산소성 허혈성 뇌손상에서 손상 후 덱사메타손의 투여 효과)

  • Park, Chang Ro;Park, Kyung Pil;Kim, Heng Mi;Sohn, Yoon Kyung
    • Clinical and Experimental Pediatrics
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    • v.46 no.10
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    • pp.989-995
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    • 2003
  • Purpose : Dexamethasone is frequently administered to prevent or treat chronic lung disease in human neonates who are also prone to hypoxic-ischemic(HI) insults. Recently, meta-analysis of the follow-up studies reveals a significantly increased odd ratio for the occurrence of cerebral palsy or an abnormal neurologic outcome, and there is conflicting evidence regarding the impact of dexamethasone exposure on HI brain injury. This study was conducted to explore the effect of post-HI dexamethasone administration on neuronal injury in neonatal rats. Methods : HI was produced in seven-day-old rats by right carotid artery ligation followed by two hours of 8% oxygen exposure. At the end of HI, the animals were injected intraperitoneally either with dexamethasone(0.5 mg/kg) or saline. Neuronal injury was assessed seven days after the HI by the area of infarction, TUNEL reactivity, Bcl-2 and Bax expression in brain. Results : Post-insult dexamethasone administration resulted in reduction of weight gain and a higher mortality rate during seven days after HI. Dexamethasone treatment revealed no effect on the size of brain infarction induced by HI. Bax protein expression increased in dexamethasone treated brain but Bcl-2 protein expression and TUNEL reactivity revealed no significant differences between dexamethasone treated and non treated brain. Increased Bax protein expression suggest upregulation of the apoptosis by dexamethasone. Conclusion : The result suggests the adverse role of Post-HI administration of dexamethasone in neonatal HI.

The Effects of Ethylacetate Fraction of Sanguisorba officinalis L. on Experimentally-induced Acute Gastritis and Peptic Ulcers in Rats (지유가 흰쥐의 급성위염 및 소화성 궤양에 미치는 영향)

  • Lee, Jae-Joon;Choi, Hyun-Suk;Lee, Jeong-Hwa;Jung, Chang-Joo;Lee, Myung-Yul
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.10
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    • pp.1545-1552
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    • 2005
  • Sanguisorba officinalis L. has been used as a traditional remedy for arthritis, neuralgia, diarrhea, vomiting, gastric ulcer and gastro-intestinal disorders. This study was performed to investigate the protective effects of ethylacetate fraction of Sanguisorba officinalis L. ethanol extract (SOE) on the gastric mucosal damage and gastric ulcer induced by HCI-ethanol, indomethacin, Shay-ligation method, restraint and water-immersion stress, and cysteamine in rats. The experiment animals were divided into four groups: a negative control group (CON), positive control group (cimetidine 100 mg/kg-CMT or omeprazol 100 mg/kg administrated group-OMT), SOE 200 mg/kg administrated group (SOL) and SOE 400 mg/kg administrated group (SOH), respectively. Rats were given an oral or intraduodenal administration of SOE, and all SOE treatment groups compared with the CON significantly inhibited HCI-ethanol-induced acute gastritis and indomethacin-induced gastric ulcers in dose-dependent manner, of which effects were lower in a positive control drug (CMT). The inhibition rates ($\%$) on the acute gastritis induced by HCI-ethanol and the gastric ulcers induced by indomethacin in rats, 17.66$\%$ and 48.28$\%$ in SOL, 21.71$\%$, and 51.08$\%$ in SOH, and 47.26$\%$ and 58.26$\%$ in CMT compared with CON, respectively, In pylorus ligated rats, the groups of SOE showed decreases in the volume of gastric secretion and acid output. However, no significant differences were observed in the pepsin activity between treated groups . In addition, SOH also depressed gastric ulcers induced by restraint and water-immersion stress and duodenal ulcers induced by cysteamine. These results suggest that Sanguisorba officinalis L. has remarkable antigastric ulcer effects and could be developed as a new antigastric ulcer agents.

Na, K-ATPase Activity in the Aged Erythrocytes of Hypertensive Rats (고혈압쥐 노화 적혈구에서의 Na, K-ATPase에 관한 연구)

  • Park, Chang-Kil;Hur, Gang-Min;Seok, Jung-Ho;Lee, Jae-Heun
    • The Korean Journal of Pharmacology
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    • v.27 no.1
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    • pp.33-43
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    • 1991
  • To study the age dependent change of Na, K-ATPase in the erythrocyte of hypertensive rat, 1-kidey 1-clip hypertensive rat was made by the removal of right kidney and partial ligation of left renal artery. After 4 weeks, aged erythrocyte fraction was separated by density gradient centrifugation, and Na, K-ATPase activity and $^3H-ouabain$ binding with ghost cell membrane and ouabain sensitive Rb-uptake with whole cell were measured. 1) In the hypertensive rats, blood pressure was significantly increased to 165.5/119.0 mmHg (systolic/diastolic). Mean corpuscular volume and membrane protein(mg) per $10^9RBC$ were decreased and hemoglobin content was increased in the aged erythrocyte. 2) Na, K-ATPase activity in the solution containing 110 mM NaCl and 10 mM KCI, was decreased in hypertensive rat, and decreased in aged erythrocyte of both group. 3) Ouabain sensitive Rb-uptake by low RbCl concentration(4 mM) was slightly decreased in aged erythrocyte compared to that in young erythrocyte of each group, but slightly increased in young erythrocyte in hypertensive rat compared to that in normotensive rat. 4) Ouabain sensitive Rb-uptake by high RbCl concentration(16 mM) was decreased about 30% to 50 % in aged erythrocyte in both group. And in hypertensive rat, especially in young erythrocyte it was significantly decreased compared to that in normotensive rats. 5) $^3H-ouabain$ binding at 0.13 or $1{\times}10^-6M$ ouabain concentration was slightly decreased in aged erythrocyte of normotensive rat, and significantly decreased in aged erythrocyte of hypertensive rats. 6) $^3H-ouabain$ binding at 6 or $64{\times}10^-6M$ ouabain concentration is slightly decreased in aged erythrocyte of both group, but significantly decreased in young and aged erythrocyte of hypertensive rats compared to that of normotensive rats. The present results suggest that (1) in the young erythrocyte of hypertensive rat, the alterations of Na-pump activity that slightly increased in weak stimulation and inhibited in strong stimulation, may be related to increased molecular activity and the decrease in the number of low affinity site without change in high affinity site, (2) in the aged erythrocyte of normotensive rat, inhibited Na-pump may be related to the change in molecular activity of pump. (3) And in the aged erythrocyte of hypertensive rat, it may be related to the decrease in the number of high and low affinity site as well as the change in molecular activity

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Clinic Analysis of Heart Surgery -110 cases- (심장수술 110례 임상고찰)

  • Chang, Hoon;Shin, Sung-Aia;No, Joong-Kee;Rho, Jun-Ryang
    • Journal of Chest Surgery
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    • v.34 no.8
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    • pp.597-603
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    • 2001
  • Background: The purpose of writing this article is to get better clinical results and further clinical improvement based on subject to 110 cases of cardiac surgery which were performed and clinically analyzed. Material and Method: Since January 1995, the patent ductus arteriosus surgery had started in our hospital. In February 1999, an open heart surgery had started and up to September 2000, total of 110 cases were performed as of double ligation of patent ductus arteriosus(10 cases) and open heart surgery(100 cases). Result: Among the patients, Korean-Chines was 74(67.3%) and Han-Chinese was 35(31.8%). Congenital heart disease was 95 cases and acquired valvular heart disease was 15 cases. 83 cases of acyanotic congenital heart disease consisted of ventricular septal defect(VSD) with associated anomaly(45 cases), atrial septal defect(ASD) with associated anomaly(20 cases), patent ductus arteriosus(PDA) with associated anomaly(11 cases), cogenital aortic stenosis(5 cases), double chamber right ventrical(1 case) and Ebstein's anomaly(1 case). Among the 12 cases of cyanotic congenital heart disease, 11 cases of tetralogy of Fallot underwent total correction. Among the 15 cases of acquired valvular heart disease, valvular replacement(7 cases), double valve replacement (3 cases), mitral valve replacement(3 cases) and aortic valve replacement(1 case) were performed. And 8 cases of valvuloplasty were performed by using of commissurotomy, chordal plasty, plasty of papillary muscle, ring type annuloplasty, repair of leaflet. Conclusion: on cogenital heart disease, short term results of surgery for acyanotic cogenital heart disease was good. Among the cyanotic congenital heart disease, tetralogy of Fallot showed a little difference of recovery according to the surgery method so that further follow up observation was needed for long term result. On acquired valvular heart disease, especially, in terms of short term result of valvuloplasty, was relatively good, but further follow up observation was also needed for long term result. There wasn's any operative mortality.

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Neuroprotection of Recombinant Human Erythropoietin Via Modulation of N-methyl-D-aspartate Receptors in Neonatal Rats with Hypoxic-ischemic Brain Injury (신생 백서의 저산소성 허혈성 뇌손상에서 NMDA receptor 조절을 통한 유전자 재조합 인 에리스로포이에틴의 신경보호)

  • Jang, Yoon-Jung;Seo, Eok-Su;Kim, Woo-Taek
    • Neonatal Medicine
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    • v.16 no.2
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    • pp.221-233
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    • 2009
  • Purpose: Erythropoietin (EPO) has neuroprotective effects in many animal models of brain injury, including hypoxic-ischemic (HI) encephalopathy, trauma, and excitotoxicity. Current studies have demonstrated the neuroprotective effects of EPO, but limited data are available for the neonatal periods. Here in we investigated whether recombinant human EPO (rHuEPO) can protect the developing rat brain from HI injury via modulation of NMDA receptors. Methods: In an in vitro model, embryonic cortical neuronal cell cultures from Sprague-Dawley (SD) rats at 19-days gestation were established. The cultured cells were divided into five groups: normoxia (N), hypoxia (H), and 1, 10, and 100 IU/mL rHuEPO-treated (H+E1, H+ E10, and H+E100) groups. To estimate cell viability and growth, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay was done. In an in vivo model, left carotid artery ligation was performed on 7-day-old SD rat pups. The animals were divided into six groups; normoxia control (NC), normoxia Sham-operated (NS), hypoxia-ischemia only (H), hypoxia-ischemia+vehicle (HV), hypoxia-ischemia+rHuEPO before a HI injury (HE-B), and hypoxia-ischemia+rHuEPO after a HI injury (HE-A). The morphologic changes following brain injuries were noted using hematoxylin and eosin (H/E) staining. Real-time PCR using primers of subunits of NMDA receptors (NR1, NR2A, NR2B, NR2C and NR2D) mRNA were performed. Results: Cell viability in the H group was decreased to less than 60% of that in the N group. In the H+E1 and H+E10 groups, cell viability was increased to >80% of the N group, but cell viability in the H+E100 group did not recover. The percentage of the left hemisphere area compared the to the right hemisphere area were 98.9% in the NC group, 99.1% in the NS group, 57.1% in the H group, 57.0% in the HV group, 87.6% in the HE-B group, and 91.6% in the HE-A group. Real-time PCR analysis of the expressions of subunits of NMDA receptors mRNAs in the in vitro and in vivo neonatal HI brain injuries generally revealed that the expression in the H group was decreased compared to the N group and the expressions in the rHuEPO-treated groups was increased compared to the H group. Conclusion: rHuEPO has neuroprotective property in perinatal HI brain injury via modulation of N-methyl-D-aspartate receptors.