표피하 수포성 질환의 진단에서 표피 추출물 면역전사법과 Bullous Pemphigoid 180 NC16a 면역전사법의 유용성 비교

이지범 ( Lee Ji Beom ),이주영 ( Lee Ju Yeong ),윤숙정 ( Yun Sug Jeong ),원영호 ( Won Yeong Ho ),이승철 ( Lee Seung Cheol ) 대한피부과학회 2004 大韓皮膚科學會誌 Vol.42 No.1

N/A Background: The 180kD bullous pemphigoid antigen (BP180) is known to be recognized by sera from patients with bullous pemphigoid (BP), herpes gestationis (HG), and cicatricial pemphigoid (CP). A series of previous studies using BP180 recombinant proteins has shown that most sera from patients with BP and HG react with the NC16a domain of BP180. Objective: This study was performed to compare the immunoblotting results by using human epidermal extracts and by using recombinant protein of BP180 NC16a domain in the diagnosis of subepidermal bullous skin diseases. Method: We observed the reactivity with the epidermal extract of normal human foreskin and recombinant protein of BP180 NC16a domain in immunoblotting assay. Results: In immunoblotting with epidermal extract, 7(78%) and 6(67%) of the 9 BP sera reacted with BP230 and BP180, respectively. 1(50%) of the 2 HG and 2(100%) of the 2 CP sera reacted with BP180. In immunoblotting with recombinant protein of BP180 NC16a domain, 8(89%) of the 9 BP sera and 2(100%) of the 2 HG sera were immunoreactive. But, none of the CP and pemphigus sera reacted with the recombinant protein. Conclusion: The immunoblotting analysis using BP180 NC16a domain is a highly sensitive method without ambiguity in the diagnosis of subepidermal bullous skin diseases, and also is useful for the differential diagnosis of BP and HG from CP. (Korean J Dermatol 2004;42(1):23~28)

황련해독탕을 함유하는 정제 개발과 성분함량 및 약리효과 평가

이지범 ( Ji-beom Lee ),최혜민 ( Hye-min Choi ),김종범 ( Jong-bum Kim ),김정옥 ( Jung-ok Kim ),문성옥 ( Sung-ok Moon ),이화동 ( Hwa-dong Lee ) 대한본초학회 2018 大韓本草學會誌 Vol.33 No.2

Objectives : Hwangryunhaedok-tang (HRHDT) is one of the well-known prescription herbal drugs of Korean herbal medicine, which has been widely used for the treatment of various bacterial and inflammatory diseases. This study was conducted in order to develop the tablet formulations of HRHDT and compare its efficacy with the other commercial formulations. Methods : Corresponding herbal medicines comprising of HRHDT were extracted with water for 3 hr at 95~100℃ and then vacuum dried. Subsequently, some pharmaceutical excipients such as microcrystalline cellulose, croscarmellose sodium, magnesium stearate, etc were used to prepare the HRHDT tablets. The contents with characterizing components of HRHDT tablet was compared with the HRHDT decoction. The contents of characterizing components were analyzed with HPLC. Furthermore, we investigated the anti-inflammatory and anti-oxidative abilities of two different commercial HRHDT granules (HJP-1 and HJP-2) and were compared with that of the formulated HRHDT tablets. The anti-oxidant properties of HRHDR were studied using the 1,1-diphenyl-2-picryhydrazyl (DPPH) radical, contents of total flavonoid and polyphenol. In addition, based on this result the anti-inflammatory effects have verified by mechanism from LPS- treated Raw264.7 macrophages. Results : The results demonstrated that HRHDT tablets showed more anti-inflammatory and anti-oxidative effects than HJP-1, HJP-2. Moreover, it showed more superior effects in terms of dose, usability and stability than the granules. Conclusion : Hence, we concluded that in order to improve the quality and efficacy of the Korean herbal medicine, it is necessary to develop appropriate methods and establish standardized techniques for the development of good formulations.

인체 각질형성세포에서 Pityrosporum ovale에 의한 전염증성 사이토카인의 발현에 대한 연구

문현주 ( Mun Hyeon Ju ),이지범 ( Lee Ji Beom ),김성진 ( Kim Seong Jin ),이승철 ( Lee Seung Cheol ),원영호 ( Un Yeong Ho ) 대한피부과학회 2003 대한피부과학회지 Vol.41 No.11

N/A Backgrounds : Pityrosporum (P.) ovale is a member of the cutaneous microflora, and is associated with several dermatologic diseases. Little is known about the mechanism by which P. ovale causes these dermatoses, but it has been supposed to be associated with immunologic response. The epidermis is largely composed of keratinocytes, and they not only play an important structural part in forming a physical barrier to microorganisms, but also are important functionally in mediating cutaneous immune reaction by secreting various cytokines. Objectives : There have been few studies on the interaction between fungi and keratinocytes, especially the effect of P. ovale on the production of cytokines by human keratinocytes. This study is to evaluate the effects of P. ovale on pro-inflammatory cytokines such as interleukin-lα (IL-lα), tumor necrosis factor-α (TNF-α), granulocyte-macrophage colony stimulating factor (GM-CSF) producted by human keratinocytes. Methods : Normal human keratinocytes (NHK) from neonatal foreskin were suspended in culture medium with different concentrations of the P. ovale extract (0.01, 1, 10, 100㎍/㎖). The production of IL-lα, TNF-α and GM-CSF after 24 hours treatment were somiquantitated by RT-PCR. Time-course changes of the cytokines were also observed until 72 hours (0, 6, 12, 24, 48, 72 hours). NHKs were treated with P. ovale culture supernatant (0.1, 0.2, 0.5, 1.O㎖) for 24 hours, and the cytokines were measured by RT-PCR and ELISA. Results : IL-lα and GM-CSF were increased by treatment with P. ovale extract as a dose-dependent manner. But TNF-α production was not affected by the treatement. The level of IL-1α was steadily increased with its maximum at 48 hours. The level of GM-CSF was increased for 48 hours and became decreased thereafter. NHKs treated with P. ovale culture supernatant showed the increase of IL-lα and GM-CSF synthesis as a dose-dependent manner. Conclusions : P. ovale induced the pro-inflammatory cytokines in normal keratinocytes, which is supposed to induce inflammatory response by the organism in the skin. (Korean J Dermatol 2003;41(11) : 1448∼1454)

세포성 혈관지방종

김재욱 ( Kim Jae Ug ),이지범 ( Lee Ji Beom ),김성진 ( Kim Seong Jin ),이승철 ( Lee Seung Cheol ),원영호 ( Won Yeong Ho ) 대한피부과학회 2003 대한피부과학회지 Vol.41 No.8

Cellular angiolipomas are benign fatty tumors that occur as multiple subcutaneous nodules on the extremities and trunks of young adults. Although clinically benign, they may occasionally mimic Kaposi`s sarcoma or pyogenic granuloma histologically. The most important distinguishing features are encapsulation, intravascular fibrin thrombi, septation, association with other more typical angiolipomas, and occurrence in healthy individuals. A 65-year-old man presented with multiple subcutaneous nodules on the trunk and extremities for several years. Histopathologic findings showed well-circumscribed septated intradermal tumor composed of vascular endothelial cells and spindle cells forming vascular spaces intermingled with scattered adipocytes, consistent with the findings of cellular angiolipoma. (Korean J Dermatol 2003;41(8) : 1065~1067)

선천성 멜라닌 세포성 모반 154예에 대한 임상 및 병리 조직학적 연구

김재욱 ( Kim Jae Ug ),이지범 ( Lee Ji Beom ),김성진 ( Kim Seong Jin ),이승철 ( Lee Seung Cheol ),원영호 ( Won Yeong Ho ) 대한피부과학회 2003 대한피부과학회지 Vol.41 No.8

N/A Background : Congenital melanocytic nevus(CMN) is regarded as benign, however it can be transformed to malignant melanoma. Thus, careful follow-up is mandatory. Objective : To identify the clinical and histologic characteristics of CMN. Method : We evaluated the clinical characteristics of CMN from 154 patients diagnosed at the Department of Dermatology in Chonnam National University Hospital from January, 1993 to December, 2002. Histopathologic exam was done from 50 patients. Results : Female was more common(62.3%) and most cases(96.8%) were clinically apparent at birth. Medium-sized nevi(1.5cm≥, <20cm) was most common(75.3%). The commonest location was on the head and neck region(45.5%). Most lesions were brown, dark brown or black in color, and flat-surfaced or slightly elevated in contour. About half of the cases manifested excessive hair growth on the surface and the possibility to have hair was positively correlated with the size of CMN. Some cases accompanied with neurofibromatosis, nevus flammeus and vitiligo were observed. Large CMN, compared with medium-sized or small CMN, involved deeper in the dermis. In 25 cases(50%), nevus cells involved the skin appendages, nerves, and vessels. Conclusion : Medium-sized CMNs on the head and neck were most prevalent. Some cases were associated with other diseases, especially pigmentary and vascular disorders. There was positive correlation between the size of CMN and the possibility to be hairy and, larger CMN tended to involve deeper in lower reticular dermis. (Korean J Dermatol 2003;41(8) : 1028~1033)

가미소요산의 새로운 제형에 대한 항산화 활성 및 항염증 효능평가

최혜민 ( Hye-min Choi ),김세진 ( Se-jin Kim ),김인수 ( In-su Kim ),이지범 ( Ji-beom Lee ),김종범 ( Jong-beom Kim ),문성옥 ( Sung-ok Moon ),이화동 ( Hwa-dong Lee ) 대한본초학회 2016 大韓本草學會誌 Vol.31 No.6

Objectives : Gamisoyosan (GMS) is a useful prescription for treating insomnia, dysmenorrhea and infertility induced by a stress. Also, GMS has been used traditionally to improve systemic circulation and biological energy production. The purpose of this study was to assess the anti-oxidant activity and anti-inflammatory effects of Gamisoyosan Formulation (Soft extract, GMS-SE). Methods : The biological activities such as anti-oxidant and anti-inflammatory effects were measured through cell line-based in vitro assay. We investigated the anti-oxidant properties of GMS-SE on the 1,1-diphenyl- 2-picryhydrazyl (DPPH) radical, contents of total flavonoid and polyphenol. GMS-SE compared to butyl hydroxy anizole (BHA). Furthermore, based on this result the anti-inflammatory effects of GMS-SE have verified by mechanism from LPS- treated Raw264.7 macrophages. Results : The anti-oxidant activities of GMS-SE increased markedly, in a dose-dependent manner. The GMS-SE showed significant scavenging activity (GMS-SE 500 ㎍/㎖ : 32.77±1.65%, GMS-SE 1000 ㎍/㎖ : 45.06±1.04% and BHA 100 ㎍/㎖ : 39.25±2.41% for DPPH assay). and, The total phenolic compound and flavonoids contents of GMS-SE were 73.93±6.87 ㎍/㎎ and 698.75±6.78 ㎍/㎎. GMS-SE which is LPS has diminished in the LPS-induced release of inflammatory mediators (NO, iNOS, COX2 and PGE2) and pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β) from the RAW264.7 macrophages. Moreover, GMS-SE inhibited the activation of phosphorylation of p38 and ERK MAPKs by induced LPS. Conclusion : The present results indicate that GMS-SE has an anti-oxidant and anti-inflammatory properties, therefore may be beneficial in diseases which related to oxidative stress-mediated inflammatory disorders.

림프종과 동반된 치료에 반응하지 않은 전신성 습진

임주영 ( Im Ju Yeong ),윤숙정 ( Yun Sug Jeong ),이지범 ( Lee Ji Beom ),김성진 ( Kim Seong Jin ),원영호 ( Won Yeong Ho ),이승철 ( Lee Seung Cheol ) 대한피부과학회 2004 대한피부과학회지 Vol.42 No.3

The common causes of eczema are known as contactant, drug, infection, atopy, and so on. Although various clinical manifestations of paraneoplastic syndrome have been described, especially in leukemia or lymphoma, intractable eczema is rarely reported to be associated with malignancy. A 55-year-old female presented with a 3-month history of pruritic erythematous papules and plaques. Oozing on the scalp, face, neck, both inguinal and inframammary areas were seen. Histopathological studies revealed a nonspecific subacute spongiotic dermatitis. The patient had been treated with prednisolone, antihistamines and antibiotics, but the skin lesions were aggravated. During the early course of eczema, laboratory findings were within the normal limits except for mild leukocytosis. Three months later, however, severe leukocytosis and cervical and inguinal lymphadenopathies developed abruptly. Computed tomography (CT) of the neck, chest, and abdomen revealed multiple lymphadenopathies to suggest lymphoma, which was confirmed later by lymph node and bone marrow biopsies, showing atypical lymphocytes extended to the bone marrow. This case shows that generalized eczema recalcitrant to conventional therapy may be associated with underlying lymphoma or leukemia (Korean J Dermatol 2004;42(3):313~316)

대시호탕의 새로운 제형이 3T3-L1에서 지방세포 증식과 분화 과정에 미치는 영향

최혜민 ( Hye-min Choi ),김세진 ( Se-jin Kim ),문성옥 ( Sung-ok Moon ),이지범 ( Ji-beom Lee ),이하영 ( Ha-young Lee ),김종범 ( Jong-beom Kim ),이화동 ( Hwa-dong Lee ) 대한본초학회 2018 大韓本草學會誌 Vol.33 No.2

Objectives : Daesiho-tang (DSHT) has been widely used in the treatment of cerebral infarct in traditional medicine. However, there was not report on the anti-obesity-related diseases efficacy of DSHT. In this study, we investigated the effects for the new formulation of DSHT, on the adipocyte differentiation cycle in 3T3-L1 cells. Methods : 3T3-L1 cells were treated with DSHT (50, 100, 200 ㎍/㎖) during differentiation for 6 days. Also, the inhibitory effect of DSHT against 3T3-L1 adipogenesis was evaluated in various stage of adipogenesis such as early (0-2day), intermediate (2-4day), and terminal stage (4-6day). The accumulation of lipid droplets was determined by Oil Red O staining. and, the expressions of genes related to adipogenesis were measured by RT-PCR and Western blot analyses. Results : DSHT showed inhibitory activity on adipocyte differentiation at 3T3-L1 preadipocytes without affect cell toxicity as assessed by measuring fat accumulation and adipogenesis. In addition, DSHT significantly reduced the expression levels of several adipocyte marker genes including proliferator activated receptor-γ (PPAR-γ) and CCAAT/ enhancer-binding protein-α (C/EBP-α). Also, the anti-adipogenic effect of DSHT was strongly limited in the intermediate (2-4 day), terminal stage (4-6 day) of 3T3-L1 adipogenesis. In addition, the DSHT treatment down- regulated mRNA expression levels of PPAR-γ,, C/EBP-α in mature 3T3-L1 adipocytes. Conclusions : These results suggest that, the ability of DSHT has inhibited overall adipogenesis and lipid accumulation in the 3T3-L1 cells. The new formulation of DSHT may be a promising medicine for the treatment of obesity and related metabolic disorders.

갈근탕 혼합단미엑스제 및 전탕액의 성분 및 생리활성 비교 연구

김정옥(Jung Ok Kim),김세진(Se Jin Kim),김종범(Jong Beom Kim),남원희(Won Hee Nam),이지범(Ji Beom Lee),이화동(Hwa Dong Lee) 한의병리학회 2019 동의생리병리학회지 Vol.33 No.1

구아바·녹차잎 추출복합물(ILS-F-2301)의 기능성 원료 표준화를 위한 지표성분 Guaijaverin과 Epigallocatechin Gallate의 분석법 개발 및 검증

전영희(Young-Hee Jeon),박유진(Yu Jin Park),박세호(Se-Ho Park),김진수(Jin Soo Kim),이지범(Ji-Beom Lee) 한국식품영양과학회 2023 한국식품영양과학회지 Vol.52 No.12

본 연구에서는 HPLC를 이용하여 구아바・녹차잎 추출복합물(ILS-F-2301)을 기능성 원료로 개발하기 위한 원료 표준화의 일환으로 ILS-F-2301의 지표성분을 guaijaverin과 EGCG로 설정하고 이에 대한 HPLC 분석법을 확립하여 유효성 검증을 실시하였다. Guaijaverin과 EGCG의 분석을 위해 0.1% formic acid가 첨가된 물과 메탄올을 이동상으로하고 C18 column을 사용하여 기울기 용리법으로 분리하고 각 물질의 최대흡수파장을 고려하여 254 ㎚(guaijaverin)와 280 ㎚(EGCG)에서 검출하였다. 분석법 유효성 검증은 특이성, 직선성, 정확성 및 정밀성 등을 통해 확인하였다. 표준용액과 ILS-F-2301의 피크 머무름시간과 UV spectrum이 일치하였으며, 다른 물질과 간섭없이 피크가 분리된 것으로 나타나 특이성을 확인하였다. Guaijaverin과 EGCG 검량선의 결정계수(R2)는 0.990 이상으로 높은 직선성을 보였으며, 검출한계는 guaijaverin이 0.44 ㎍/mL, EGCG가 21.44 ㎍/mL였고 정량한계는 guaijaverin이 1.32 μg/mL, EGCG가 64.97 ㎍/mL로 확인되었다. ILS-F-2301에 표준물질을 3개 농도로 첨가하고 분석한 회수율은 guaijaverin이 100.17~104.86%, EGCG가 97.92~99.40%로 나타나 정확성이 있는 것을 확인하였다. ILS-F-2301의 정밀도 측정 결과, guaijaverin과 EGCG의 상대표준편차(RSD)는 각각 0.39~1.38%, 0.16~0.52%로 나타나 본 분석법은 정밀성이 있음을 확인하였다. 이상의 결과로 ILS-F-2301의 지표성분인 guaijaverin과 EGCG의 HPLC 분석법이 신뢰성 있음을 검증하였다. 따라서 본 연구를 통해 확립된 시험법이 향후 ILS-F-2301의 원료 또는 제품의 품질관리를 위한 기초 자료로 활용될 것으로 기대된다. This study validated an analytical method using HPLC for the standard determination of guaijaverin and epigallocatechin gallate (EGCG) in mixed extracts of Psidium guajava and Camellia sinensis (ILS-F-2301) as functional ingredient. The analytical method of guaijaverin and EGCG was optimized by HPLC on a C18 column with a gradient elution of 0.1% aqueous formic acid and methanol at a flow rate of 0.8 mL/min. The analyte was detected at 254 and 280 ㎚. Validation of an analytical method was evaluated with respect to specificity, linearity, accuracy, and precision. Guaijaverin and EGCG were successfully separated with identical RT and UV spectrum without any interference substance, demonstrating that it presented specificity. Calibration curve of guaijaverin and EGCG exhibited high linearity (R²≥0.990). The limit of detection and quantitation of guaijaverin and EGCG were 0.44, 1.32 ㎍/mL and 21.44, 64.97 ㎍/mL, respectively. The recovery rate of guaijaverin and EGCG were obtained 100.17∼104.86% and 97.92∼99.40%, respectively, indicating that the result exhibited high accuracy. Relative standard deviation values in precision of guaijaverin and EGCG were 0.39∼1.38% and 0.16∼0.52%, respectively, showing that it appeared the excellent precision. Taken together, these results suggest that the developed HPLC method for the analysis of guaijaverin and EGCG in ILS-F-2301 is simple, efficient, and reliable. Also, it could be contributed to the quality control of ILS-F-2301.