CD28Ig 및 CTLA4Ig 융합단백 생산 포유동물 세포주 수립에 관한 연구

유양석,정용훈,최용,임대철,조양자 한양대학교 의과대학 1998 한양의대 학술지 Vol.18 No.2

CTLA-4, which is homologous to CD28, recognizes the same coutner-receptors B7.1 and B7.2. Although initial researches implied that these molecules may act to each other synergistically, recent data suggest that CTLA-4 may acts as a negative regulator of T cell activation. In this study human CD28 and CTLA-4 cDNAs were amplified from PHA-stimulated human peripheral blood lymphocytes mRNAs by using RT-PCR method. IgG1 Fc portion cDNA was amplified from human peripheral blood lymphocytes of patients with febrile illness and fused to C"-terminal parts of CD28 and CTLA-4 extracellular domains. The fusion constructs were subcloned into pCI-neo vector and then transfected into Sp-2/0murine myeloma cell. Stable cell lines were established by geneticin selection and cloning. In the sequence of CTLA-4 cDNA from 3 different healthy persons it was found that 2 sites of bases were different from that published in original paper and submitted to genbank database. The 49th and 331st bases of open reading frame were changed from adenine to guanine and from guanine to adenine respectively. It was confirmed that the 17th and 111th amino acid of human CTLA-4 were alanine and threonine respectively, 5 different human CTLA-4 clones with serial N'-terminal deletion were constructed. CTLA4/L_1 was intact CTLA-4 gene of native form without any form of mutation. CTLA4/L_2 was a mutatn with deletion in gene coding N'-termial 6 amino acids. CTLA4/L_3 was a mutant with deletion in gene coding N'-termial 11 amino acids. CTLA4/L_4 was a mutant with deletion in gene coding N'-termial 16 amino acids. CTLA4/L_5 was a mutant with deletion in gene coding N'-termial 22 amino acids. Level of surface expression of CTLA4Ig on L_1, L_2, L_3, L_4 and L_5 transfectant cell lines were 5%, 3%, -1.5%, 8%, and 6% respectively. These 2 fusion proteins, CD28Ig and CTLA4Ig, would be useful tools for further researches on role of the costimulations in T cell activation.

치면열구전색제의 조건에 따른 법랑질 결합강도에 관한 연구

이용근,이흥수,이성재,방기숙,최정민,류승호,강민석 대한치과기재학회 1997 대한치과재료학회지 Vol.24 No.1

In this study, the shear bond strengths and accompanying elastic modulus ratio, displacement at bond failure and work up to fracture of pit and fissure sealants to toothe enamel were analyzed by the following variables, those were acid etching, moisture content of the surface of enamel, dentin bonding agent(species and method), thermocycling, saliva contamination. One kind of chemically cured resin based pit and fissure sealant, two kinds of light cured resin based pit and fissure sealants and one kind of glass ionomer based pit and fissure sealant, and two kinds of dentin bonding agents were studied. The buccal surface of extracted premolars were ground flat with # 600 silicone carbide papers, and the ground surface was treated depending on the experimental conditions. After each treatment, each pit and fissure sealant was filled into plastic tube of 3mm diameter, which were laid on the surface of ground enamel surface, and were cured. Sealant bonded teeth specimens were immersed in 37℃ distilled water form 24 hours. The shear bond strength of sealant to enamel surface was measured with Universal testing machine with the cross head speed of 1mm/min. From the obtained load-deflection curves, modulus ratio, bond strength, displacement at bond failure and work up to fracture were analyzed. From this experiment, the following results were obtained : 1. Comparing the results of standard condition(i. e. acid etching, dry surface, no dentin bonding agent and 300 thermocyling), the elastic modulus ratio of chemically cured resinbased pit and fissure sealant was significantly higher than those of other materials(P<0.05), and the bond strength, displacement at bond failure and work up to fracture of glass ionomer based pit and fissure sealant were significantly lower than those of other materials(P<0.05). 2. The results of ANOVA with the variables of type of material, acid etching, moisture condition of the enamel surface, dentin bonding agents were as follows ; 1) variables showed significant main effect on the shear bond strength of pit and fissure sealant to enamel(P<0.01), 2) type of material, acid etching and moisture condition of enamel influenced the bond strength significantly(P<0.01), 3) type and application methods of dentin bonding agents influenced on the bond strength significantly(P<0.05). 3. There were statistically significant correlation between the work up to fracture and the bond strength, and between the work up to fracture and the displacement at bond failure(P<0.01), and in these cases the correlation coefficient was 0.87.

둔부 통증을 주소로 내원한 괴사성 근막염 1례

안승찬,류석용,이상래,조석진,오성찬,김홍용 인제대학교 2008 仁濟醫學 Vol.29 No.-

Necrotizing fasciitis (NF) is a rare, life-threatening infection resulting in necrosis of the skin, subcutaneous tissue, and fascia. A very severe and usually fatal fasciitis is caused by a virulent species of streptococcus that is often referred to as the "flesh-eating bacteria". Mortality rates have been noted high. Certain conditions can predispose patients to NF, such as diabetes mellitus, immunosuppressive medications, and AIDS. Patients usually complain of excessive pain as well as constitutional symptoms. Because of this rapid progression, it is important to diagnose and treat NF quickly to decrease mortality. Treatment includes broad-spectrum antibiotic coverage, nutritional supplements, hemodynamic support, wound care, and prompt surgical debridement. We report a case of a 51-year-old man who had presented with right hip pain. Previously, he had been diagnosed with diabetes mellitus and alcoholic liver cirrhosis.

3차 의료기관 응급실로 내원한 안면부 열상 환자에 대한 임상적 고찰

김양원,안성훈,류석용,김홍용,전병민,김기태 대한응급의학회 2001 대한응급의학회지 Vol.12 No.2

Background: Facial laceration is a common clinical problem in the emergency room that is frequently associated with other craniofacial injuries. It elicits an emotional response from the patient because of the underlying concern of permanent scar and secondary facial disfigurement, so they demand prompt and adequate treatment. The aim of this study is to analyze the incidence, etiology, depth, and site of facial lacerations to provide basic data for further understanding. Method: This study reviewed 1,043 facial-laceration patients treated in the emergency room at Pusan Paik Hospital between March 1999 and February 2000. This retrospective study was done by reviewing and analyzing the sex and age distributions, the monthly and daily distributions, the causes of injury, the types of injury, the sites of injury, the lengths of the lacerations, and associated injuries. Results: The sex ratio of men to wemen was 2.4:1, and the 0~10 age group was at the top of the age distribution. The number of patients was the highest on sundays and during March. About 9.2% of the patients visited the emergency room between 10:00 P.M. and 11:00 P.M. Falls(38.1%) were the most common cause of facial laceration, and deep laceration(51.8%) was the most common type of facial laceration. The most common laceration site was the forehead, followed by the chin and the nose. The most common associated craniofacial injury was facial bone fracture(39%), Facial-lacerations were sutured by plastic surgeons(80%), emergency physicians(10%), and oromaxillofacial surgeons(10%). We found statistical significance in the types and lengths of lacerations between men and women, among age groups(p<0.05). Conclusion: Facial laceration is one of the most common craniofacial injuries in the emergency room. By now, most lacerations have been repaired by plastic surgeons, But, the causes of lacerations are diverse and many patients are admitted due to the associated injuries. Therefore, emergency physicians should participate more aggressively in the care of facial-laceration patients to improve the outcome of the patients.

자발성 심막기종 1례

안지영,이상래,안성훈,류석용,김홍용 대한응급의학회 2001 대한응급의학회지 Vol.12 No.3

Pneumopericardium, as a form of barotrauma, refers to the presence of air within the pericardial sac. The causes of pneumopericardium are various, Clinically, pneumopericardium is typically present with dyspnea and precordial chest pain. On physical examination, heart sounds are usually distant, and precordial tympany may be elicited. The diagnosis can be made by clinical and radiographic findings. Management of pneumopericardium depends on many factors, such as the age of the patient, the suspected causes, and the extent of clinically observed respiratory compromise. The case shown here, a patient who developed pneumopericardium, involved an associated underlying pulmonary parenchymal process as the cause of pneumopericardium.

고삼투압성 비케톤성 상태의 예후인자

안성훈,김양원,김미란,진헌철,안지영,이상래,류석용,김홍용,김성준,이병권,김경환 대한응급의학회 2001 대한응급의학회지 Vol.12 No.2

Background: A hyperosmolar nonketotic state has been known to have a high mortality, and even now, despite this high mortality, only a few studies of this disease have been performed. We studied the prognostic factors for the hyperosmolar nonketotic state. Methods: We retrospectively studied the cases of 40 patients who were in a hypersomolar nonketotic state when admitted to Sanggye Paik Hospital during the 6-year Period from 1995 through 2000. We divided the hyperosmolar nonketotic patients into two groups, the complete recovery group and the incomplete recovery group, and compared the clinical features, the laboratory findings, and the precipitating factors between two groups. Results: 1) A total of 40 patients were studied: 24 in the complete recovery group and 16 patients in the incomplete recovery group. The mortality rate was 32.5%. 2) No significant statistical difference existed among the clinical features of the two groups, except for the sex(p<0.01). 3) Among the laboratory findings of both groups, analysis revealed that the effective osmolarity was significantly higher among those in the incomplete recovery group(p<0.01). Serum sodium concentration was also significantly higher among those in the incomplete recovery group(p<0.01). Serum creatinine was also significantly higher among those in the incomplete recovery group(p<0.05). Serum bicarbonate concentration, on the other hand, was significantly lower among those in that group(p<0.05).4) Infection was identified as the most common precipitating factor(62.5%). Among the precipitating factors of the two groups, there were significant statistical difference in pneumonia, UTI, and inappropriate glucose control. 5) A significant statistical difference existed among the initial level of consciousness of both groups(p<0.05). 6) The only significant independent factor responsible for prognosis of nonketotic hyperosmolar state patients was the sex. Conclusion: The sex was only significant independent prognostic factor of nonketotic hyperosmolar state patients.

Inflammatory Cytokine Expression and Sebum Production after Exposure of Cultured Human Sebocytes to Ultraviolet A Radiation and Light at Wavelengths of 650 nm and 830 nm

( Weon Ju Lee ),( Soo Yuhl Chae ),( Hyo Sub Ryu1 ),( Yong Hyun Jang ),( Seok Jong Lee ),( Do Won Kim ) 대한피부과학회 2015 Annals of Dermatology Vol.27 No.3

Background: The effectiveness of ultraviolet (UV) radiation, visible light, or infrared light therapy for the treatment of acne is the subject of ongoing scientific debate. Objective: This study was conducted to investigate changes in sebum production and the expression of inflammatory cytokines, matrix metalloproteinases (MMPs), and antimicrobial peptides (AMPs), following exposure of cultured human sebocytes to UVA radiation and light at wavelengths of 650 nm and 830 nm. Methods: Reverse transcription polymerase chain reaction assays were performed to measure the gene expression levels of inflammatory cytokines (interleukin [IL]-1β, IL-6, IL-8, and tumor necrosis factor-α), MMPs (MMP-1, MMP-3, and MMP-9), and AMPs (psoriasin, hBD-2, hBD-3, and LL-37) in cultured sebocytes after exposure to UVA radiation (2 J/cm2, 3 J/cm2, and 5 J/cm2) and light at wavelengths of 650 nm (14 J/cm2, 29 J/cm2, and 87 J/cm2) and 830 nm (5 J/cm2, 10 J/cm2, and 30 J/cm2). Expression of inflammatory cytokine proteins and sebum production were measured using enzyme- linked immunoassays and a lipid analysis kit, respectively. Results: Exposure of cultured sebocytes to UVA radiation and light at wavelengths of 650 nm and 830 nm did not show a significant increase in the expression of inflammatory cytokines, MMPs, or AMPs. Sebum production was not significantly decreased after exposure to UVA radiation and light at both wavelengths. Conclusion: We propose that UVA radiation, visible light, and infrared light can be used to target Propionibacterium acnes for the treatment of acne, without an increase in the expression of inflammatory biomarkers and sebum production. (Ann Dermatol 27(2) 163∼ 170, 2015)

생리식염수 및 포도당수액 정액 투여가 에탄올 농도에 미치는 효과에 관한 연구

김원율,이상래,안지영,안성훈,류석용,김경환,김홍용 대한응급의학회 2000 대한응급의학회지 Vol.11 No.3

Background: In emergency department, there are many drunken patients with various emergency medical situations. In these patients, evaluation and management of the medical problems are delayed from time to time because of the uncooperative nature of the patients and because of consciousness and sensory changes. A precise evaluation can only be obtained when the patients become sober. For this purpose, most clinicians start IV fluid loading. This study was carried out to evaluate the effect of IV fluid(normal saline and glucose solution) on the rate of ethanol clearance in such patients Methods: Ten volunteers(healthy males in their twenties and with a Michigan alcohol screening test below 4) were enrolled in this study. Alcohol intake was restricted to at least 48 hours before the test and food intake to at least 4 hours. For the test, A predetermined dose of ethanol(1 gm/kg) was given to the volunteers. Each volunteer was tested on three consecutive times with 3 or more days interval for alcohol wash out period . On the lst day, the volunteers received ethanol only per os. On the second day, they received ethanol and a liter of IV 10% dextrose solution. Each day, blood was drawn from the antecubital vein to measure the ethanol level at the time of ethanol ingestion and at 0.5, 1, 1.5, 2, 3, 4, 5 and 6 hours after ingestion. The repeated measure ANOVA test was used for the statistical analysis. Results: The difference in the blood ethanol level between the 3 test was F=1.7, p=0.184 and the difference corrected by the time factor was F=0.32, and p=0.985. Conclusion: There was no significant difference in the blood ethanol level between the tests. In conclusion, the IV normal saline or glucose solution does not accelerate ethanol clearance in suffering

Cnidicin, a Coumarin, from the Root of Angelica koreana, Inhibits the Degranulation of Mast Cell and the NO Generation in RAW 264.7 Cells

Ryu, Shi Yong,Kou, Na Youn,Choi, Hong Seok,Ryu, Hyun,Kim, Tae Sung,Kim, Kyeong Man 전남대학교 약품개발연구소 2001 약품개발연구지 Vol.10 No.-

Cnidicin (1) and five related coumarins were isolated from the root extract of Angelica koreana (Umbelliferae) as active principles responsible for the inhibitory effect on the degranulation process of cultured mast cells. Cnidicin (1) demonstrated a significant inhibition upon the release of β-hexosaminidase from the cultured RBL-2H3 cells in a dose dependent manner (IC_50 value, 25μM) and upon the nitric oxide production from the activated RAW264.7 cells (IC_50 value, 7.5μM). In agreement with this, cnidicin inhibited the expression of nitric oxide synthase in RAW264.7 cells.

STIM1 Regulates Somatic Ca²⁺ Signals and Intrinsic Firing Properties of Cerebellar Purkinje Neurons

Ryu, Changhyeon,Jang, Dong Cheol,Jung, Dayoon,Kim, Yong Gyu,Shim, Hyun Geun,Ryu, Hyun-Hee,Lee, Yong-Seok,Linden, David J.,Worley, Paul F.,Kim, Sang Jeong Society for Neuroscience 2017 The Journal of neuroscience Vol.37 No.37

<P>Control of Ca2+ flux between the cytosol and intracellular Ca2+ stores is essential for maintaining normal cellular function. It has been well established in both neuronal and non-neuronal cells that stromal interaction molecule 1 (STIM1) initiates and regulates refilling Ca2+ into the ER. Here, we describe a novel, additional role for STIM1, the regulation of free cytosolic Ca2+, and the consequent control of spike firing in neurons. Among central neurons, cerebellar Purkinje neurons express the highest level of STIM1, and they fire continuously in the absence of stimulation, making somatic Ca2+ homeostasis of particular importance. By using Purkinje neuron-specific STIM1 knock-out (STIM1(PKO)) male mice, we found that the deletion of STIM1 delayed clearance of cytosolic Ca2+ in the soma during ongoing neuronal firing. Deletion of STIM1 also reduced the Purkinje neuronal excitability and impaired intrinsic plasticity without affecting long-term synaptic plasticity. In vestibulo-ocular reflex learning, STIM1(PKO) male mice showed severe deficits in memory consolidation, whereas they were normal in memory acquisition. Our results suggest that STIM1 is critically involved in the regulation of the neuronal excitability and the intrinsic plasticity of the Purkinje neurons as well as cerebellar memory consolidation.</P>