약용버섯의 계통분류 및 국내유통 Inonotus속내 종간 구별을 위한 신속동정법 개발

김성윤 외 한국버섯학회 2005 한국버섯학회지 Vol.3 No.2

국내유통약용버섯의 분류체계를 확립하고 이들 속 및 종간의 유연관계확립을 위하여 계통학적 정보를 지니고 있는 ITS부위의 염기서열을 밝히고 ITS1과 ITS2부위의 다양한 염기서열을 이용하여 분류학적 위치를 확립하였고, 시판 I. obliquus 종의 진위여부와 계통분류학적인 유연관계 확립 및 종 특이적인 유전자 탐침을 개발하였다. 본 연구의 조사결과에 의하면, 약용버섯으로 주로 시판되고 있는 국내유통균주는 총 6개의 속(Phellinus, Inonotus, Sparassis, Fomes, Ganoderma, Hericium)으로 나누어짐을 알 수 있었고 그 중에서 기존에 잘 알려진 상황버섯과 최근 들어 수입양이 급증하고 있는 차가버섯이 대부분을 차지하고 있는 것으로 확인되었다. 본 연구에 사용되어진 일명 차가버섯으로 유통 중인 15개 제품 중 중국에서 수입되어진 59번 균주가 P. pini로 확인되었으며 일본에서 수입되어진 51, 52번 균주가 P. baumii와 P. linteus와 유사종 혹은 동일종으로 확인되었다. 한편 I. rheades(AY237731)와 I. radiatus(AY354217) 및 F. fomentarius(AY354213)는 NCBI 등록 시 문제점이 있는 것으로 사료되며 본 실험에서 조사되어진 30번 균주 F. fomentarius가 정확한 말굽버섯인 것으로 사료된다. The Aphyllophorales is a large order containing about 2,000 known species. Many of these are the bracket and coral fungi. The vast majority of these fungi are saprophytic on the plant debris. Many species are significant in decomposing plant remains, as they are able to digest cellulose or lignin that occurs in plant cell walls. Many of these fungi have been involved in everyday human affairs. A few were used medicinally by the Greeks and Romans as a remedy for many complaints, including colic, fractured limbs and bruises. Other bracket fungi have been used as curry combs for horses, as snuff, as razor strops and as a source of dye for clothing. The texture of the basidiocarp may be similar to that of cork, wood, leather, paper, or cartilage. Unlike the basidiocarps of the Order Agaricales, the basidiocarps of the Aphyllophorales are not fleshly and moist. Division of the members of the Aphyllophorales into genera was originally made on the basis of gross morphology of the basidiocarp and hymenium and Donk(1964) recognizes 22 families in this order. The species and genus whose typical in Aphylloporales were listed in Table. with related information. The ITS region sequence of some genus were found by BLAST search. Sequences retrieved from GenBank were visually aligned by the program CLUSTAL G. As a result, the medicinal mushroom was separated in four groups. In this multiple alignment, the sequence analysis among Fomes group, Inonotus group and Phellinus group showed high genetic similarity except Hericium group and Sparassis group.

Three Cases of Spontaneous Intracranial Hypotension (SIH) Treated with Epidural Blood Patch

신진우,윤창섭,이 철 대한통증학회 1997 The Korean Journal of Pain Vol.10 No.1

Spontaneous intracranial hypotension, a syndrome of low CSF pressure, occurs without preceding events such as lumbar puncture, back trauma, operative procedure or medical illness. The most char- acteristic symptom is an occipital or frontal headache which is aggrevated in the erect position and relieved in the supine position. This syndmme usually resolves spontaneously or with strict bed rest. When the headach persists or is incapacitating, more aggressive treatment may be necessary. Autologous epidural blood patch is highly effective in the management of spontaneous intracranial hypotension. Epidural blood produces an organized clot which effectively tamponade any dural CSF leak The rapid relief of headache immediately after the infusion of blood occur by some other mechanism, such as an increase in subarachnoid pressure that is known to occur with infusion of fluid into the lumbar epidural Space. We report three cases of spontaneous intracranial hypotension successfully treated with epidural blood patch.

Induction of Apoptosis by Aqueous Extract of Cordyceps militaris Through Activation of Caspases and Inactivation of Akt in Human Breast Cancer MDA-MB-231 Cells

Jin, Cheng-Yun,Kim, Gi-Young,Choi, Yung-Hyun The Korean Society for Microbiology and Biotechnol 2008 Journal of microbiology and biotechnology Vol.18 No.12

Cordyceps militaris is well known as a traditional medicinal mushroom and has been shown to exhibit immunostimulatory and anticancer activities. In this study, we investigated the apoptosis induced by an aqueous extract of C. militaris (AECM) via the activation of caspases and altered mitochondrial membrane permeability in human breast cancer MDA-MB-231 cells. Exposure to AECM induced apoptosis, as demonstrated by a quantitative analysis of nuclear morphological change and a flow cytometric analysis. AECM increased hyperpolarization of mitochondrial membrane potential and promoted the activation of caspases. Both the cytotoxic effect and apoptotic characteristics induced by AECM treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, which demonstrates the important role of caspase-3 in the observed cytotoxic effect. AECM-induced apoptosis was associated with the inhibition of Akt activation in a time-dependent manner, and pretreatment with LY294002, a PI3K/Akt inhibitor, significantly increased AECM-induced apoptosis. The results indicated that AECM-induced apoptosis may relate to the activation of caspase-3 and mitochondria dysfunctions that correlate with the inactivation of Akt.

TRAIL attenuates sulforaphane-mediated Nrf2 and sustains ROS generation, leading to apoptosis of TRAIL-resistant human bladder cancer cells

Jin, Cheng-Yun,Molagoda, Ilandarage Menu Neelaka,Karunarathne, Wisurumuni Arachchilage Hasitha Maduranga,Kang, Sang-Hyuck,Park, Cheol,Kim, Gi-Young,Choi, Yung Hyun Elsevier 2018 Toxicology and applied pharmacology Vol.352 No.-

<P><B>Abstract</B></P> <P>Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) can preferentially initiate apoptosis in malignant cells with minimal toxicity to normal cells. Unfortunately, many human cancer cells are refractory to TRAIL-induced apoptosis through many unknown mechanisms. Here, we report that TRAIL resistance can be reversed in human bladder cancer cell lines by treatment with sulforaphane (SFN), a well-known chemopreventive isothiocyanate in various cruciferous vegetables. Combined treatment with SFN and TRAIL (SFN/TRAIL) significantly induced apoptosis concomitant with activation of caspases, loss of mitochondrial membrane potential (MMP), Bid truncation, and induction of death receptor 5. Transient knockdown of <I>Bid</I> prevented collapse of MMP induced by SFN/TRAIL, consequently reducing apoptotic effects. Furthermore, SFN increased both the generation of reactive oxygen species (ROS) and the activation of nuclear factor erythroid 2-related factor 2 (Nrf2), which is an anti-oxidant enzyme. Interestingly, TRAIL effectively suppressed SFN-mediated nuclear translocation of Nrf2, and the period of ROS generation was more extended compared to that of treatment with SFN alone. In addition, silencing of <I>Nrf2</I> increased apoptosis in cells treated with SFN/TRAIL; however, blockade of ROS generation inhibited apoptotic activity. These data suggest that SFN-induced ROS generation promotes TRAIL sensitivity and SFN can be used for the management of TRAIL-resistant cancer.</P> <P><B>Highlights</B></P> <P> <UL> <LI> SFN sensitizes TRAIL-mediated apoptosis in TRAIL-resistant bladder cancer cells. </LI> <LI> SFN-mediated TRAIL sensitization requires Bid cleavage, leading to apoptosis. </LI> <LI> SFN enhances DR5 expression, resulting in cell death. </LI> <LI> SFN/TRAIL increases ROS generation in spite of Nrf2 activation. </LI> <LI> TRAIL helps to sustain SFN-mediated ROS generation. </LI> </UL> </P>

Bcl-2 Overexpression Attenuates Agaricus blazei Murill-Induced Apoptosis in U937 Cells by Inhibition of Caspase-3 Activit

Cheng-Yun Jin,Gi-Young Kim,Jae-Yun Lee,Mun-Ok Kim,Jin-Young Park,Dong-Oh Moon,Yung-Hyun Choi,Jae-Dong Lee 한국버섯학회 2006 한국버섯학회지 Vol.4 No.2

Agaricus blazeiMurill is an edible mushroom distributed in Brazil and presently cultivated in other areas, including Korea, Japan, and China. Its chemical components, including steroids and lectin and various polysaccharides have been widely studied. For this, we used U937/vector and U937/Bcl-2 cells, which were generated by transfection of the cDNA of the Bcl-2 gene. As compared with U937/vector, U937/Bcl-2 cells exhibited a 4-fold greater expression of Bcl-2. Treatment with 0.5 or 4 mg/ml A. blazei Murill for 24 h produced morphological features of apoptosis in U937/vector cells, respectively. This was associated with caspase-3 activation and PARP degradation. In contrast, A. blazei Murill-induced caspase-3 activation and PARP degradation and apoptosis were significantly inhibited by z-DEVD-fmk in U937 cells. Bcl-2 overexpressing cells exhibited sustained caspase-3 activation and expression levels of the Bcl-2 proteins during A. blazei Murill-induced apoptosis. In addition, these findings indicate that Bcl-2 inhibits A. blazei Murill-induced apoptosis by a mechanism that interferes with Bcl-2 degradation and activity of caspase-3 that is involved in the execution of apoptosi.

Bcl-2 and Caspase-3 Are Major Regulators in <i>Agaricus blazei</i>-Induced Human Leukemic U937 Cell Apoptosis through Dephoshorylation of Akt

Jin, Cheng-Yun,Moon, Dong-Oh,Choi, Yung Hyun,Lee, Jae-Dong,Kim, Gi-Young Pharmaceutical Society of Japan 2007 Biological & pharmaceutical bulletin Vol.30 No.8

<P><I>Agaricus blazei</I> is a medicinal mushroom that possesses antimetastatic, antitumor, antimutagenic, and immunostimulating effects. However, the molecular mechanisms involved in <I>A. blazei</I>-mediated apoptosis remain unclear. In the present study, to elucidate the role of the Bcl-2 in <I>A. blazei</I>-mediated apoptosis, U937 cells were transfected with either empty vector (U937/vec) or vector containing cDNA encoding full-length Bcl-2 (U937/Bcl-2). As compared with U937/vec, U937/Bcl-2 cells exhibited a 4-fold greater expression of Bcl-2. Treatment of U937/vec with 1.0—4.0 mg/ml of <I>A. blazei</I> extract (ABE) for 24 h resulted in a significant induction of morphologic features indicative of apoptosis. In contrast, U937/Bcl-2 exposed to the same ABE treatment only exhibited a slight induction of apoptotic features. ABE-induced apoptosis was accompanied by downregulation of antiapoptotic proteins such as X-linked inhibitor of apoptosis protein (XIAP), inhibitor of apoptosis protein (cIAP)-2 and Bcl-2, activation of caspase-3, and cleavage of poly(ADP-ribose)polymerase (PARP). Ectopic expression of Bcl-2 was associated with significantly induced expression of antiapoptotic proteins, such as cIAP-2 and Bcl-2, but not XIAP. Ectopic expression of Bcl-2 also reduced caspase-3 activation and PARP cleavage in ABE treated U937 cells. Furthermore, treatment with the caspase-3 inhibitor z-DEVD-fmk was sufficient to restore cell viability following ABE treatment. This increase in viability was ascribed to downregulation of caspase-3 and blockage of PARP and PLC-γ cleavage. ABE also triggered the downregulation of Akt, and combined treatment with LY294002 (an inhibitor of Akt) significantly decreased cell viability. The results indicated that major regulators of ABE-induced apoptosis in human leukemic U937 cells are Bcl-2 and caspase-3, which are associated with dephosphorylation of the Akt signal pathway.</P>

Induction of apoptosis through down-regulation of AKT and activation of Caspase-3 in Human breast cancer MDA-MB-231 cells by aqueous extract of Codyceps militaris

Cheng-Yun Jin,Mun-Ock Kim,Jin-Woo Jeong,Jae-Dong Lee 한국버섯학회 2007 한국버섯학회지 Vol.5 No.3·4

Cordyceps militaris is well known as a traditional herbal ingredient, which has been used for patients suffering from cancer in oriental medicine. In this study we have investigated the biochemical mechanisms of anti-proliferative effects by C. militaris extract(CBE) in human breast cancer MDA-MB-231 cells. It was found that CBE treatment induced chromatin condensation, mitochondrial energization, annexin V staining and sub-G1 phase DNA content. These indicators of apoptosis correlate with the mitochondrial dependent pathway, which results in the activation of caspase-3 activity. Both the cytotoxic effect by CBE treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, demonstrating the important role of caspase-3 in the observed cytotoxic effect. Co-treatment of CBE and LY294002, resulted in significantly induction of apoptosis. These results indicate that caspase-3 is a key regulator of apoptosis in response to CBE in human breast cancer MDA-MB-231 through down regulation of Akt, and that the C. militaris extract may therefore have therapeutic potential against human breast cancer.

Oleifolioside B-mediated autophagy promotes apoptosis in A549 human non-small cell lung cancer cells

JIN, CHENG-YUN,YU, HAI YANG,PARK, CHEOL,HAN, MIN HO,HONG, SU HYUN,KIM, KYOUNG-SOOK,LEE, YOUNG-CHOON,CHANG, YOUNG-CHAE,CHEONG, JAEHUN,MOON, SUNG-KWON,KIM, GI-YOUNG,MOON, HYUNG-IN,KIM, WUN-JAE,LEE, JAI- Spandidos Publications 2013 International journal of oncology Vol.43 No.6

Sarijang Enhances Maturation of Murine Bone Marrow-Derived Dendritic Cells

Cheng-Yun Jin(김성윤),Min Ho Han(한민호),Cheol Park(박철),Hye Jin Hwang(황혜진),Eun-A Choi(최은아),Yung Hyun Choi(최영현) 한국생명과학회 2011 생명과학회지 Vol.21 No.12

수지상세포(DCs)는 항원을 섭취하여 말초조직에서 lymphoid 기관으로 이동하는 특화된 항원제시세포(APCs)로서 미성숙 T 세포를 자극함으로서 일차적 면역반응에 중심적인 역할을 하기 때문에 DC의 성숙에 대한 조절은 면역학적 치료 접근에 매우 중요한 부분이다. 본 연구에서는 서목태 발효 산물이 주 원료인 사리장에 의한 DC의 성숙 유도 가능성을 조사하기 위해 GM-CSF와 IL-4를 이용하여 골수 유래 수지상세포(BMDCs)를 대상으로 DC의 성숙에 관여하는 주요 인자들의 발현에 미치는 영향을 LPS 처리군과 비교하였다. 사리장은 처리농도 의존적으로 표면 수용체인 CD80 및 CD86의 발현을 증가시켰으나, CD80 발현 증가에 더 유의적이었다. 또한 사리장은 MHC I 보다 MHC II의 발현을 현저하게 증가시켰으며, 이러한 결과는 사리장이 DC의 성숙을 위한 적용에 매우 유의적으로 사용될 수 있을 가능성과 면역활성 효능을 가질 수 있음을 의미하는 것이다. Dendritic cells (DCs) are professional antigen-presenting cells playing key roles in immune sentinels as initiators of T-cell responses against microbial pathogens and tumors. Sarijang, a folk sauce containing extracts of Rhynchosia nulubilis, Ulmus davidiana roots, Allium sativum, and Rhus Verniaiflura bark, has been used as a nonspecific immunostimulant for cancer patients. However, little is known about its immunomodulating effects or their mechanisms. In this study, we investigated whether sarijang induces phenotypic and functional maturation of DCs. For this study, murine bone marrow-derived myeloid DCs were cultured in the presence of interleukin-4 (IL-4) and granulocyte-macrophage colony stimulating factor (GM-CSF), and the generated immature DCs were stimulated with sarijang or lipopolysaccharide (LPS). Our data indicated that sarijang significantly enhanced the expression of co-stimulatory molecules (CD80 and CD86) as well as major histocompatibility complex (MHC) II, as did LPS. The results provide new insight into the immunopharmacology of sarijang and suggest a novel approach to the manipulation of DC for therapeutic application.

Curcumin attenuates the release of pro-inflammatory cytokines in lipopolysaccharide-stimulated BV2 microglia

JIN, Cheng-yun,LEE, Jae-dong,PARK, Cheol,CHOI, Yung hyun,KIM, Gi-young Springer Science and Business Media LLC 2007 Acta pharmacologica Sinica. Vol.28 No.10