• Biomolecules & Therapeutics
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• v.8 no.2
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• pp.153-159
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• 2000

A novel serine/threonine protein phosphatase with EF-hand motif, which belongs to PPEF family was partially cloned from rat brain cDNA by employing RT-PCR method. The size of the amplified clone was 1.6kbp. The amplified DNA was subcloned into pGEM-T-Easy vector and the resulting plasmid was maned as pGEM-rPPEF2. The nucleuotide sequence is shared by 88% with that of mouse PPEF-2 cDNA, and the deduced amino acid sequence reveal 92% homology with that of mouse PPEF-2 cDNA. The N-terminal region of the cloned rat brain PPEF contains a putative phosphatase catalytic domain (PP domain) and the C-terminal region contains multiple $Ca^{2+}$ binding sites (EF region). The putative catalytic domin (PP) and the EF-hand motif (EF) regions were subcloned into pGEX4T-1 and were overexpressed in E. coli DH5 as glutathione-S-transferase (GST) fusion proteins. Expression of the desired fusion protein was identified by SDS-PAGE and also by immunoblot analysis using monoclonal antibody against GST. The recombinant proteins were purified by glutathione-agarose chromatography. This report is first to demonstrate the cloning of PPEF family from rat brain tissues. The clone reported here would be invaluable for the investigation of the role of this new type-phosphatase in rat brain.

• Biomolecules & Therapeutics
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• v.22 no.3
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• pp.246-253
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• 2014

Codonopsis lanceolata has been used as an herbal medicine for several lung inflammatory diseases, such as asthma, tonsillitis, and pharyngitis. Previously, we showed the neuroprotective effect of steamed and fermented C. lanceolata (SFC) in vitro and in vivo. In the current study, the treatment of HT22 cells with SFC decreased glutamate-induced cell death, suggesting that SFC protected HT22 cells from glutamate-induced cytotoxicity. Based on these, we sought to elucidate the mechanisms of the neuroprotective effect of SFC by measuring the oxidative stress parameters and the expression of Bax and caspase-3 in HT22 cells. SFC reduced contents of ROS, $Ca^{2+}$ and NO. Moreover, SFC restored contents of glutathione and glutathione reductase as well as inhibited Bax and caspase-3 activity in HT22 cells. These results indicate that steamed and fermented C. lanceolata (SFC) extract protected HT22 cells by anti-oxidative effect and inhibition of the expression of Bax and caspase-3.

• BMB Reports
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• v.29 no.6
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• pp.555-563
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• 1996

A membrane-bound phosphatidylinositol 4-kinase (PI 4-kinase) was separated in a sucrose gradient and solubilized with 1% Triton X-100 from mouse brain. The enzyme was purified 2,952-fold by various chromatographic techniques including DEAE-cellulose, PI-Sepharose and Sephacryl S-200 gel filtration. The molecular weight of PI 4-kinase was approximately 76 kDa by gel filtration and 70.8 kDa by SDS-polyacrylamide gel electrophoresis. The purified enzyme exhibited specific activity of 11.2 nmol/min/mg protein and pi value of 4.7. Kinetic analysis of the PI 4-kinase indicated apparent $K_m$, values of 190 ${\mu}M$ and 120 ${\mu}M$ for phosphatidylinositol and ATP, respectively. The maximal activity of this purified enzyme was observed at pH 7.4 at an incubation temperature of $37^{\circ}C$. The enzyme activity was significantly activated by $Mg^{2+}$, $Mn^{2+}$ and $Fe^{2+}$, and inhibited severely by $Ca^{2+}$. PI 4-kinase was proved to be pure in its immunoblot test by polyclonal antibody prepared from immunized rabbit sera. By this test, we were able to detect the existence of the same type of PI 4-kinase from other mouse organ tissues, such as liver, heart, kidney and spleen. Furthermore, similar immunoblot analysis with the same antisera recognized the different epitopes of PI 4-kinase proteins from various organs of rabbit, chinese hamster and rat.

• Radiation Oncology Journal
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• v.9 no.2
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• pp.227-232
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• 1991

Authors described the remote afterloading endobronchial brachytherapy (EBBT technique using the microSelectron HDR Ir-192 and the Asan Medical Center experience. Total 28 EBBT in 9 patients were performed since November 1989 and 24 EBBT in 8 patients were emploiyed for palliation and 3 EBBT in 1 patient was treated curatively. Authors observed a significant relief of obstructive symptom with tumor regression in 7 patients out of 8 who were treated palliatively but one of them died of pulmonary congestion in 3 weeks after EBBT One patient with prior therapy of extensive electrocautery expired within 1 day after 2nd EBBT procedure with massive hemorrhage from the lesion. EBBT procedure has been tolerable and can be performed as an outpatient.

• Radiation Oncology Journal
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• v.15 no.3
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• pp.197-206
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• 1997

Purpose : Phospholipase D (PLD) catalyzes the hydrolysis of phosphatidylcholine to phosphatidic acid (PA) and choline. Recently, PLD has been drawing much attentions and considered to be associated with cancer Process since it is involved in cellular signal transduction. In this experiment, oleate-PLD activities were measured in various tissues of the living rats after whole body irradiation. Materials and Methods : The reaction mixture for the PLD assay contained $0.1\;\muCi\;1,2-di[1-^{14}C]palmitoyl$ phosphatidylcholine 0.5mM phosphatidylcholine, 5mM sodium oleate, $0.2\%$ taurodeoxycholate, 50mM HEPES buffer(pH 6.5), 10mM $CaCl_2$, and 25mM KF. phosphatidic acid, the reaction product, was separated by TLC and its radioactivity was measured with a scintillation counter. The whole body irradiation was given to the female Wistar rats via Cobalt 60 Teletherapy with field size of 10cmx loom and an exposure of 2.7Gy per minute to the total doses of 10Gy and 25Gy. Results : Among the tissues examined, PLD activity in lung was the highest one and was followed by kidney, skeletal muscle, brain, spleen, bone marrow, thymus, and liver. Upon irradiation, alteration of PLD activity was observed in thymus, spleen, lung, and bone marrow. Especially PLD activities of the spleen and thymus revealed the highest sensitivity toward $\gamma-rar$ with more than two times amplification in their activities In contrast, the PLD activity of bone marrow appears to be reduced to nearly $30\%$. Irradiation effect was hardly detected in liver which showed the lowest PLD activity. Conclusion : The PLD activities affected most sensitively by the whole-body irradiation seem to be associated with organs involved in immunity and hematopoiesis. This observation s1ron91y indicates that the PLD is closely related to the physiological function of these organs, Furthermore, radiation stress could offer an important means to explore the phenomena covering from cell Proliferation to cell death on these organs.

• The Journal of Korean Society for Radiation Therapy
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• v.34
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• pp.83-92
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• 2022

Purpose: To evaluate usefulness of Tomotherapy and Halcyon VMAT treatment in radiation therapy for bi-breast cancer Materials and Methods: For 10 patients with bi-breast cancer, volumetric modulated arc therapy(VMAT) treatment plan was established using helical Tomotherapy(Accuray. USA) with field width of 5.0-cm and pitch of 0.287, and Halcyon(Varian Medical System, Palo Alto, CA, Version 3.0 USA) of 6arc and 8arc. Prescribed dose was 42.4 Gy/16, and V _40.3Gy of Planning Target Volume(PTV) was 90%. The quality of plan was evaluated by comparing the dose to tumor and normal organs, and the efficiency was evaluated by comparing total MU and beam on time. Results: About three treatment plans(Tomotherapy, Halcyon 6Arc VMAT, Halcyon 8Arc VMAT) , the mean homogeneity index(H.I) of PTV were 1.07, 1.10, 1.11, and the mean conformity index(C.I) of PTV were 1.21, 1.16, 1.17, respectively. The average value of the dose assessment item for a normal organ is V _5Gy(%) of both lung was 36.3, 31.2, 29.7, and V _15Gy(%) were 18.6, 15.5, 14.6, respectively. The mean heart dose(Gy) were 4.17, 2.69, 2.51. Total MU were (7498.6, 2494.2, 2471.5), and beam on time(sec) were 462.5, 195.4, 198.0. Conclusion: Halcyon VMAT showed similar quality of treatment plan compared to helical Tomotherapy, while also protecting normal organs. In addition, the efficiency of radiotherapy increased due to a decrease in Beam On Time and MU.

• Journal of Chest Surgery
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• v.31 no.9
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• pp.827-836
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• 1998

Background: Effective cardiopulmonary resuscitation (CPR) should provide acceptable hemodynamics for the vital organs during cardiac arrest and early restoration of spontaneous circulation that guarantees long-term, neurologically intact survival. CPR using heart-lung bypass has been suggested as an option for that use. This study was designed to determine the effectiveness of standard CPR techniques, closed-vs. open-chest CPR, which could be used in the future study verifying the role of heart-lung bypass CPR. Material and Method: By using adult mongrel dogs, closed-chest CPR (CCCPR, n=4) and open-chest CPR (OCCPR, n=5) were compared with respects to hemodynamics, restoration of spontaneous circulation(ROSC), and survival. Ventricular fibrillation-cardiac arrest (VF-CA) was induced by electrical shock in all animals. After 4 minutes of cardiac arrest, basic life support (BLS) was applied for 15 minutes and followed by advanced life support (ALS). ALS was maintained until achi ving ROSC but not longer than 30 minutes regardless of the recovery. Resuscitation procedures in either group were standardized by adopting the protocol of American Heart Association. Result: Prearrest baseline hemodynamic data was not different between two groups. During resuscitation, substantially higher systolic pressure was maintained in OCCPR group than in CCCPR group (45$\pm$15 vs. 33$\pm$11 mmHg during BLS, 83$\pm$36 vs. 44$\pm$15 mmHg during ALS; p=NS). Mean pulmonary arterial pressure went up to the level of mean systemic arterial pressures in CCCPR group and to half of that in OCCPR group, and had kept higher in CCCPR group throughout CPR (32$\pm$10 vs. 22$\pm$4 mmHg during BLS and 32$\pm$15 vs. 24$\pm$10 mmHg during ALS; p=NS). ROSC was obtained in 4 of 5 dogs receiving open-chest CPR and 2 of 4 closed-chest CPR. Prolonged survival was noted in all dogs in OCCPR group (6 to 1440 hours) but not in CCPR group (p<.05). Conclusion: These findings indicate that open-chest CPR can be more effective t maintain hemodynamics during cardiac arrest and to obtain restoration of spontaneous circulation and survival. Further experiment will be designed to compare heart-lung bypass CPR with open-chest CPR.

• Childhood Kidney Diseases
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• v.5 no.2
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• pp.213-218
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• 2001

Microscopic Polyangiitis(MPA) belongs to a spectrum of systemic vasculits, and particularly antineutrophil cytoplasmic autoantibodies(ANCA)-associated small-vessel vasculitis which is characterized by involvement of the lung and kidney. The diagnosis of MPA is often difficult to make, and delayed because of the variability of the clinical presentation. Renal biopsies have a very important diagnostic and prognostic value in MPA. We experienced a case of microscopic polyangiits which was confirmed by renal biopsy and positive serum perinuclear ANCA, associated with alveolar hemorrhage and gastrointestinal bleeding. We began methylprednisolone pulse therapy, combined with a low dose of cyclophosphamide and plasmapheresis therapy. ACE inhibitor and Ca channel blocker were used when proteinuria and hypertention developed. On admission, the patient's lab findings showed BUN l17 mg/dL, Cr 2.3 mg/dL, while on the 60th hospital day BUN/Cr values fell to 20.8 mg/dL / 1.6 mg/dL and though proteinuria persisted, the patient's condition was tolerable and is currently under observation on an out-patient basis. The last lab values were BUN 26 mg/dL / Cr 1.6 mg/dL. (J. Korean Soc Pediatr Nephrol 2001 ; 5 : 213-18)

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.98-98
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• 2003

${\beta}$-(1,3)-D-Glucans have been known to exhibit antitumor and antimicrobial activities. The presence of dectin-1,${\alpha}$, ${\beta}$-glucan receptor of dendritic cell, on macrophage has been controvertial. RT-PCR analysis led to the detection of dectin-1${\alpha}$ and ${\beta}$ in murine macrophage Raw264.7 cell line. Among the various organs of mouse, dectin-1${\alpha}$ and ${\beta}$ were detected in the thymus, lung, spleen, stomach and intestine. To analyze gene expression modulated by ${\beta}$-glucan treated murine Raw264.7 macrophage, total mRNA was applied to cDNA microarray to interrogate the expression of 7,000 known genes. cDNA chip analysis showed that ${\beta}$-glucan of P. osteatus increased gene expressions of immunomodulating genes, membrane antigenic proteins, chemokine ligands, complements, cytokines, various kinases, lectin associated genes and oncogenes in Raw 264.7 cell line. When treated with ${\beta}$-glucan of P. osteatus and LPS, induction of gene expression of TNF-${\alpha}$ and IFN-R1 was confirmed by RT-PCR analysis. Induction of TNF-R type II expression was confirmed by FACS analysis. IL-6 expression was abolished by EDTA in ${\beta}$-glucan and LPS treated Raw264.7 cell line, indicating that ${\beta}$-glucan binds to dectin-l in a Ca$\^$＋＋/ -dependent manner. To increase antitumor efficacy of ${\beta}$-glucan, ginsenoside Rh2 (GRh2) was co-treated with ${\beta}$-glucan in vivo and in vitro tests. IC$\sub$50/ values of GRh2 were 20 and 25 $\mu\textrm{g}$/$m\ell$ in SNU-1 and B16 melanoma F10 cell line, respectively. Co-treatment with ${\beta}$-glucan and GRh2 showed synergistic antitumor activity with cisplatin and mitomycin C both in vitro and in vivo. Single or co-treatment with ${\beta}$-glucan and GRh2 increased tumor bearing mouse life span. Co-treatment with ${\beta}$-glucan and GRh2 showed more increased life span with mitomycin C than that with cisplatin. Antitumor activities were 67% and 72 % by co-injection with ${\beta}$-glucan and GRh2 in the absence or presence of mitomycin C, respectively.

• Tuberculosis and Respiratory Diseases
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• v.66 no.2
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• pp.127-131
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• 2009

Most mediastinal teratomas are histologically well-differentiated tumors and benign. The majority of patients with a mediastinal teratoma are asymptomatic and their tumors are usually discovered incidentally on chest radiography. On rare occasions this tumor will rupture spontaneously into the adjacent organs. A 72-year-old female patient was admitted for dyspnea and she had a multiloculated pleural effusion in the left lung field. Although repeated pleural biopsy and pleural fluid cytology did not prove the presence of malignancy, we assumed that this was a malignant effusion because it revealed consistently high levels of carcinoembryonic antigen and carbohydrate antigen 19-9, and the chest CT scan did not show typical fat or bone density in the mass. Secondary infection and an uncontrolled septic condition due to pleural empyema finally compelled the patient to undergo a surgical operation. Mature teratoma was the final diagnosis and she has done well without recurrence for 2 months.