• Clinical and Experimental Pediatrics
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• 제45권2호
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• pp.240-246
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• 2002

목 적 : 신사구체 질환의 발생과 진행에 있어서 tumornecrosis factor-${\alpha}$($TNF-{\alpha}$)의 혈청 농도가 관련이 있다는 것과 혈청 농도보다는 신장내 농도가 더 중요하다는 주장이 있다. 이에 본 연구자들은 신사구체 신염 질환인 HSP 신염(HSN)과 신증후군 질환인 특발성 신증후군(INS)에서 $TNF-{\alpha}$와 단백뇨와의 관련성을 알아보고자 했다. 방 법: 1998년 3월부터 2001년 3월까지 충북대학교병원 소아과를 방문한 HSP 21명, INS 22명을 대상으로 혈액과 소변을 채취하였다. 단백뇨가 있는 경우는 24시간 동안 단백량을 creatinine에 대한 비를 구하여 비교하였다. $TNF-{\alpha}$의 농도 측정은 mouse anti-human $TNF-{\alpha}$ 항체를 이용한 sandwitch ELISA법을 이용하였다. 통계는 Statistical Analysis System(SAS)을 이용하여 Kruskall-Wallis test, Wilcoxon rank sum test, Spearmann test, paired t-test를 시행하였으며 유의 수준은 P<0.05를 기준으로 하였다. 결 과 : HSP에서 신장 침범이 없는 환아보다 신장 침범을 보인 환아에서 $TNF-{\alpha}$ 농도는 혈청에서 높았으나(P=0.009) 요에서는 유의한 차이가 없었다(P=0.088). HSN에서 혈청 $TNF-{\alpha}$의 농도에 비례하여 요단백이 의미 있게 증가하였고(P=0.004), 요 $TNF-{\alpha}$의 농도에 따라 요단백이 증가하는 양상이었지만 통계적 의의는 없었다(P=0.053). INS에서 단백뇨와 관련하여 혈청에서 P=0.763, 요에서 P=0.007으로 요의 $TNF-{\alpha}$의 농도에서만 통계적 의의가 있었다. 결 론: HSN에서 혈청 $TNF-{\alpha}$의 농도에 비례하는 신사구체의 손상으로 인해 요단백이 증가하고, INS에서는 여과된 단백에 의한 신세뇨관의 손상과 이에 따른 요의 $TNF-{\alpha}$의 증가로 추론할 수 있다. 따라서 HSP 환아에서 초기에 혈청 $TNF-{\alpha}$의 농도를 낮추는 치료를 시행함으로서 신염으로 이환되는 것을 예방하고, HSN 환아에서도 혈청 $TNF-{\alpha}$의 농도를 낮추는 치료를 적극적으로 하여 병의 진행을 막을 수 있을 것으로 사료되며, 신증후군에서는 단백뇨를 줄이는 치료를 하여 신장의 병변이 악화되는 것을 지연시킬 수 있을 것으로 사료된다.

• 생명과학회지
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• 제30권3호
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• pp.313-319
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• 2020

최근, 국내에서 발생하는 대가축의 질병은 바이러스 혹은 세균 등과 같은 병원체가 사료 섭취, 가축 간의 신체접촉, 호흡 등 다양한 경로를 통해 전파되어 발병되는 전염성 질병이다. 전염성 질병은 가축의 건강을 위협하고 생산성을 감소시키기 때문에 현장에서 조기 진단하여 개체 격리와 같은 통제 관리가 필수적이다. 기존 사용되고 있는 진단 키트들은 현장에서 사용하기에 용이하지 않으며 극소량의 감도에서 진단이 제한적인 단점을 가지고 있다. 그러므로, 현장에서 극소량의 감도와 진단의 편이성을 고려하여 DNA와 RNA 수준에서 진단할 수 있는 CRISPR/Cas 시스템은 최적의 시스템이라 할 수 있다. 본 연구논문에서는 대가축의 전염성 질병들을 현장에서 조기 진단함에 있어 CRISPR/Cas 시스템의 활용전략에 대해 소개하고자 한다. 최근 발견된 CRISPR/Cas 효소들은 2개의 클래스와 6가지 하위유형으로 분류되었다. 이 중에서 클래스 2에 포함되는 Cas 효소들은 대표적으로 제 2형에 Cas9, 제 5형에 Cas12a와 Cas12b, 제 6형에 Cas13a와 Cas13b가 있다. 현재까지 개발된 CRISPR/Cas 시스템들은 간단한 시각 신호를 통해 표적에 대한 정량 및 다중 감지가 가능하고 특히, 극소량 수준의 초고감도에서도 표적만을 진단할 수 있으며 단시간 이내에 진단 결과를 얻을 수 있다. 하지만 초고감도 DNA 혹은 RNA를 진단하기 위해 최적의 신호 증폭 방법과 결합되어야 하고 표적 DNA 혹은 RNA를 진단에 적합하도록 DNA를 RNA로, RNA를 DNA로 전변해야 하는 단점이 있다. 따라서, 현장에서 대가축의 전염성 질병을 조기에 진단할 수 있는 CRISPR/Cas 바이오센서를 개발하는데 있어 가축의 전염 매개체로부터 추출되는 병원체 유형(DNA 혹은 RNA)을 고려하여 최적의 Cas 효소를 선정하여야 하고 이에 따른 적절한 신호 증폭 방법이 결합되어야 한다. 따라서, CRISPR/Cas 시스템은 유전자 편집 방법을 사용하는 빠르고 효율적인 진단 도구이며 이 시스템은 소의 전염병을 조기에 진단하고 감염 확산방지에 도움될 수 있을 것으로 판단 되어진다.

• 대한본초학회지
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• 제23권2호
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• pp.179-192
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• 2008

Objectives : Draconis Resina (DR) has been used as a traditional Korean herbal medicine since ancient times, and today it is used as a medication for wounds, tumors, diarrhea, rheumatism, in the itching of insect bites and with other conditions in the folk medicine. The aim of this study was to determine whether fractionated extracts of DR inhibit free radical generation, intracellular oxidation, production of nitrite, an index of NO, PGE2, iNOS, COX-2 and proinflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages, Methods : DR extract prepared with methanol, and then fractionated with hexane, dichloromethane, ethylacetate, n-butanol and water. Inhibitory effect of DR onto free radical generation was determined by measuring DPPH, superoxide anions and nitric oxide scavenging activities in vitro. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Intracelluar oxidation was analysed by DCF-DA assay. The nitric oxide (NO) production was measured by Griess reagent system. The levels of iNOS and COX-2 expression were confirmed by western blot. And proinflammatory cytokines were measured by ELISA kit. Results : Our results indicated that fractionated extracts, especially dichloromethane and ethyl acetate extracts, significantly inhibited free radical generation, the LPS-induced H202, NO, PGE2 production and iNOS, COX-2 expression accompanied by an attenuation of TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 formation in macrophages. Conclusions : Our results indicate that dichloromethane and ethyl acetate extracts of DR have potential as an agent of chronic inflammatory diseases.

• 한국콘텐츠학회논문지
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• 제9권12호
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• pp.391-399
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• 2009

본 연구의 목적은 중추신경계 손상의 쥐에 운동강도에 따른 신경계 회복을 알아보기 위해 부하수영과 무부하수영이 혈청 BDNF 농도와 행동학적 변화에 어떠한 영향을 미치는가를 확인하고자 하였다. 실험동물로는 Sprague-Dawley 수컷 흰쥐를 사용하였으며, Ketamine을 복강 내에 주사하여 마취한 뒤 제 1-2요추의 척수에 6-OHDA를 $100{\mu}{\ell}$ 주입하여 척수손상을 유발시켰다. 척수 손상으로 유발된 행동학적 변화를 알아보기 위해 BBB와 경사판 검사를 기록하였다. 혈청 BDNF는 수술 후 14일에 혈액을 채취해 흡광도를 측정하였다. 척수 손상 후 무부하군에서는 행동변화가 대조군에 비해 유의하게 증가하였고, 혈청 BDNF 농도는 다른 군에 비해 무부하군에서 증가하였다. 이러한 결과로 보아 저강도 수영운동을 중추신경 손상 후 초기에 적용하면 신경조직의 회복에 영향을 주리라 생각된다.

• Clinical and Experimental Reproductive Medicine
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• 제30권1호
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• pp.65-75
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• 2003

Objectives: To investigate the role of TGF (Transforming growth factor-$\beta$) involved in the paracrinic communication during decidualization between UEC (uterine epithelial cells) and USC (uterine stromal cells), we have employed a co-culture system composed of human endometrial epithelial and stromal cells in defined hormonal conditions. Design: In the co-culture, endometrial epithelial cells cultured in the matrigel-coated cell culture insert are seeded on top of the endometrial stromal cells cultured within a collagen gel. The co-culture was maintained for 48 hours under the following hormonal conditions: progesterone dominant condition (100 nM P4 and 1 nM E2) or estrogen-dominant condition (100 nM E2 and 1 nM P4). 10 ng/ ml HGF and/or 10 ng/ml TGF-$\beta$1 are added. Methods: RT-PCR is utilized to detect mRNAs quantitatively. Enzyme-linked immunosorbent assay (ELISA) and immunohistochemical staining are utilized to detect proteins in the tissue. Results: Prolactin mRNA is expressed in the co-cultured stromal cells under the progesterone dominant condition. TGF-$\beta$1 and its receptors are expressed in both the co-cultured epithelial and stromal cells irrespective of the steroid present, which is in contrast with no or negligible expression of TGF-$\beta$1 or its receptor in cells separately cultured. Both estrogen and progesterone significantly elevate the concentration of hepatocyte growth factor (HGF) in the conditioned medium of the co-culture with the value of 4, 325 pg/ml in E2-dominant and 2, 000 pg/ml in P4-dominant condition compare to 150 pg/ml in no hormone. In separately cultured stromal cells, administration of HGF induces the expression of TGF receptor 1 in both hormonal conditions, but induction of TGF receptor 2 is only manifest in the P4-dominant condition. Administration of TGF-$\beta$ and HGF directly induce the decidualization marker prolactin mRNA in separately cultured stromal cells. Conclusion: It is likely that steroid hormones induces prolactin mRNA indirectly by promoting the cell to cell communication between the stromal and the epithelial cells. TGF-$\beta$ and HGF are two possible paracrine mediators in the human endometrial decidualization.

• 생물정신의학
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• 제12권2호
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• pp.189-195
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• 2005

Objects:Some studies have suggested that brain-derived neurotrophic factor(BDNF), one of the most important neurotrophins, is involved in pathophysiology of depression and suicide. This study was performed to determine whether there is an abnormality in plasma BDNF concentration in suicidal attempters. Methods:The subjects were 71 suicidal attempters who visited emergency rooms in multi-medical centers. All subjects had been interviewed by using Structured Clinical Interview for DSM-IV(SCID), Hamilton Depression Rating Scale(HDRS), Young Mania Rating Scale(YMRS), and Positive And Negative Syndrome Scale(PANSS). The severity of the suicidal behavior was measured by Lethality of Suicide Attempt Rating Scale(LSARS) and Risk-Rescue Rating(RRR) system. Seventy-one age, sex, and diagnosis matched non-suicidal psychiatric patients who were consecutively admitted to a psychiatric ward during the same period recruited as psychiatric controls. They were drug-naive or drug-free at least more than 2 months. In addition, 80 healthy controls were randomly selected as normal controls. Plasma BDNF level was measured by the enzyme linked immunosorbent assay(ELISA) methods. Results:In overall F-test, differences of the plasma BDNF levels among the groups were statistically significant(F=20.226, p<0.001). In the multiple comparisons(Scheffe), while mean levels of plasma BDNF between normal controls and non-suicidal psychiatric patients were similar(p=0.984), the BDNF levels of suicidal attempters were lower than those of other two groups(p<0.001). LSARS and RRR did not reveal any significant correlations with BDNF levels in suicidal attempters. Conclusion:These results suggest that reduction of plasma BDNF level is related to suicidal behavior and BDNF level may be a biological marker of suicidal behavior.

• 동의생리병리학회지
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• 제16권2호
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• pp.233-238
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• 2002

Moxibustion is one of major healing technique in oriental medicine. It has been widely used in many diseases such as rheumatoid arthritis, Hashimoto disease, breech presentation, etc. However, till now, effects of moxibustion on NK cell activity and relations between sympathetic nerve system(SNS) and the immune alteration induced by moxibustion were not well studied. This study was designed to evaluate effects of moxibustion on NK cell activity and the intervention of SNS in the alteration of NK cell activity induced by moxibustion. Splenic NK cytotoxity was measured in a standard 4-h 51Cr release assay. We measured the NK cytotoxity at after moxibustion stimulation for 1,3,5, and 7 days, and also measured the NK cell cytotoxity after 3 and 7 days burn stimulation with similar temperature. IL-2, IL-4, INF-γ in serum were measured by rat IL-2, IL-4, INF-γ ELISA TEST KIT. To evaluate the effects of sympathectomy on alteration of NK cell cytotoxity, 6-hydroxydopamine(6-OHDA : 5Omg/kg) was used. We showed that NK cell activity of moxibustion stimulation group increased at the 3rd day, and declined at 7th day in comparison with that of contol group. In moxibustion stimulation group, NK cell activity of 3 day stimulation group was significantly higher than sham group. On the contrary, in burn stimulation group, NK cell activity was significantly higher than that of sham groups at 3rd, 7th days. Patterns between moxibustion and burns were different. INF- γ level of 3 days moxibustion stimulation group significantly higher than sham group. IL 2 level among groups were not different. IL-4 was not detected in serum with this method. Sympathectomy abolished the NK cell activity alteration induced by moxibustion. The results suggest that moxibustion induces the alteration of NK cell activity, along with INF-γ and SNS is related to these effects.

• 약학회지
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• 제48권6호
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• pp.328-334
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• 2004

Cervical cancer is one of the leading causes of female death. Viral oncoproteins E6 and E7 are selectively retained and expressed in carcinoma cells infected with HPV (Human pa pilloma virus) type 16 and cooperated in immotalization and transformation of primary keratinocyte. E6 and E7 oncoproteins interfere the functions of tumor suppressor proteins p53 and retinoblasoma protein (pRb), respectively. Among a lots of natural products, Mentha arvensis Linne var.piperascens have inhibitory effects on bindings between E6 oncoprotein and tumor suppressor p53, E3 ubiqutin- protein ligase (E6AP). HPV oncoprotein inhibitors from Mentha piperita L. were isolated by solvent partition and column chromatography (Silica gel, RP-18) and inhibitory compounds were finally purified by HPLC using an ELISA screening system based on binding between E6 and E6AP. The aim of this study is to identify the structure of inhibitory compounds and to investigate whether these compounds have inhibitory effects on functions of E6 oncoprotein. We investigated whether caffeic acid methyl ester (CAM) extracted from Mentha piperita L. could inhibit the function of E6 oncoprotein. CAM inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53 and also inhibited the proliferation of human cervical cancer cell lines (SiHa and CaSKi) in a dose response manner. These results suggest that CAM inhibited the function of E6 oncoprotein, suggesting that it can be used as a potential drug for the treatment of cervical cancers infected with HPV.

• 대한치과보철학회지
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• 제46권3호
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• pp.307-319
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• 2008

STATEMENT OF PROBLEM: It is known that an anodic oxidation technique, one of the methods for the implant surface treatment, remarkably increased surface area, enhanced wettability and accelerated the initial bone healing. Purpose: This study was performed to evaluate the wettability of anodized titanium surface which has a nanotubular structure, to assess osseointegration after the placement of implant with nano-size tubes on tibia of rats and to analyze quantitatively transferable rhBMP-2 on each surface. MATERIAL AND METHOD: Four different kinds of surface-treated titanium discs (polished (machined surface) group, micro (blasting surface) group, nano (anodizedmachined surface) group, and nano-micro (anodized-blasting surface) group) were fabricated (n=10). Three different media were chosen to measure the surface contact angles; distilled water, plasma and rhBMP-2 solution. After a single drop (0.025 $m{\ell}$) of solution, the picture was taken with the image camera, and contact angle was measured by using image analysis system. For the test of osseointegration, 2 kinds of anodized surface (anodized-machined surface, anodized-blasting surface) implants having 2.0 mm in diameter and 5.0 mm in length inserted into the tibia of Wistar rats. After 3 weeks, tibia were harvested and the specimens were stained with hematoxylin and eosin for histological analysis. To test the possibility of drug delivery, after soaking sample groups in the concentration of 250 ng/$m{\ell}$l of rhBMP-2 for 48 hours, the excess solution of rhBMP-2 were removed. After that, they were lyophilized for 24 hours, and then the rhBMP-2 on the surface of titanium was resolved for 72 hours in PBS. All the extracted solution was analyzed by ELISA. One-way analysis of variance (ANOVA) was performed on the data. RESULTS: The wettability is improved by anodic oxidation. The best wettability was shown on the nano-micro group, and it was followed by nano group, micro group, and polished group. In the histological findings, all implants showed good healing and the new bone formation were observed along the implant surface. After 3 days, nano-micro group delivered the most amount of rhBMP-2, followed by nano group, micro group, and polished group. CONCLUSION: It indicated that anodic oxidation on blasting surface produce functionally graded nano-micro porous structure and enhance hydrophilicity of the surface and osseointegration. The findings suggest that the nano-micro porous structure could be a useful carrier of osteogenic molecules like rhBMP-2.

• Asian Pacific Journal of Cancer Prevention
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• 제15권17호
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• pp.7169-7174
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• 2014

Background: NPAS2 is a product of the circadian clock gene. It acts as a putative tumor suppressor by playing an important role in DNA damage responses, cell cycle control and apoptosis. Chronic lymphocytic leukemia (CLL) appears to be an apoptosis related disorder and alteration in the NPAS2 gene might therefore be directly involved in the etiology of CLL. Here, the Ala394Thr polymorphism (rs2305160:G>A) in the NPAS2 gene was genotyped and melatonin concentrations were measured in a total of seventy-four individuals, including thirty-seven CLL cases and an equal number of age- and sex-matched healthy controls in order to examine the effect of NPAS2 polymorphism and melatonin concentrations on CLL risk in a Pakistani population. Materials and Methods: Genotyping of rs2305160:G>A polymorphism at NPAS2 locus was carried out by amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). Melatonin concentrations were determined by enzyme linked immunosorbent assay (ELISA). Statistical analysis was performed using Statistical Package for Social Sciences software. Results: Our results demonstrated no association of the variant Thr genotypes (Ala/Thr and Thr/Thr) with risk of CLL. Similarly, no association of rs2305160 with CLL was observed in either females or males after stratification of study population on a gender basis. Moreover, when the subjects with CLL were further stratified into shift-workers and non-shift-workers, no association of rs2305160 with CLL was seen in either case. However, significantly low serum melatonin levels were observed in CLL patients as compared to healthy subjects (p<0.05). Also, lower melatonin levels were seen in shift-workers as compared to non-shift-workers (p<0.05). There was no significant difference (p>0.05) in the melatonin levels across NPAS2 genotypes in all subjects, subjects with CLL who were either shift workers or non-shift-workers. General Linear Model (GLM) univariate analysis revealed no significant association (p>0.05) of the rs2305160 polymorphism of the NPAS2 gene with melatonin levels in any of the groups. Conclusions: While low melatonin levels and shift-work can be considered as one of the risk factors for CLL, the NPAS2 rs2305160 polymorphism does not appear to have any association with risk of CLL in our Pakistani population.