Multiplex-PCR을 이용한 제주지역 소아청소년의 급성설사질환 역학조사

이규택 ( Kyu-taeg Lee ),김선미 ( Sunmi Kim ),정무상 ( Moo Sang Chong ) 대한임상검사과학회 2017 대한임상검사과학회지(KJCLS) Vol.49 No.2

2015년 3월부터 2017년 2월까지 2년간 Multiplex-PCR을 이용한 소아청소년의 급성설사질환 원인규명을 위해 의뢰된 521 분변 검체를 대상으로 5종의 바이러스 검사와 10종의 세균검사를 실시한 521 분변 검체 중 바이러스나 세균이 확인된 170명 중 중복 감염이 확인된 9명의 검체를 포함하여 179개의 양성검체의 결과를 분석한 결과 남아가 102명(56.98%), 여아가 77명(43.02%)이었으며, 3∼4세(51.96%), 5∼6세(12.29%) 연령대에서 가장 높은 양성율을 보였다. 179건의 양성 검체 중 중복감염을 포함한 209건(209/521, 40.12%)을 분석한 결과 norovirus-GII 감염이 88건(88/521, 16.89%), Campylo-bacter spp. 감염이 26건(26/521, 4.99%), rotavirus 감염이 18건(18/521, 3.45%), Clostridium difficile Toxin B 감염이 18건(18/521, 3.45%), adenovirus 감염이 17건(17/521, 3.26%), Clostridium perfringens 감염은 16건(16/521, 3.07%), astrovirus 감염은 11건(11/521, 2.11%), Salmonella spp. 감염은 5건(5/521, 0.96%), norovirus-GI, Yersinia spp., Aeromonas spp. 감염은 각각 3건(3/521, 0.58%), verocy-totoxin-producing E. coli 감염은 1건(1/521, 0.19%), Vibrio spp, E. coli O157:H7, Shigella spp. 감염은 나타나지 않았다. 유아기(6세 이하)의 계절별 분리 양상을 확인한 결과는 기온이 낮은 겨울(11∼2월)에는 주로 norovirus-GI, norovirus-GII가 유행하였고, 기온이 상승하기 시작하는 봄(3∼5월)에는 rotavirus가 유행하는 경향을 전형적인 장염바이러스의 경향을 보였다. Astrovirus 경우에는 norovirus-GI, norovirus-GII, rotavirus가 유행하는 시기가 아닌 4월∼10월에 유행하는 경향을 보이고, adenovirus인 경우에는 8월∼2월까지 유행하는 경향을 보였다. 연령이 높아질수록 바이러스뿐만 아니라 세균이 원인이 되어 급성설사질환을 유발하는 것으로 나타났으며, 다중 중합효소연쇄반응으로 원인을 파악하고자 할 때에는 유아기 때에는 바이러스 검사를 의뢰하는 것이 유효하지만 연령이 높아짐에 따라 바이러스와 세균을 동시에 의뢰하는 것이 바람직하다고 사료된다. To examine the cause of acute diarrheal disease in children and adolescents, 521 fecal samples underwent multiple-PCR for a period of two years, between March 2015 and February 20178, in the Jeju region of Korea. Based on the analysis of 179 positive samples, 102 samples were male (56.98%) and 77 were female (43.02%), and highest positive rates were shown in the age group of 3∼4 years (51.96%) and 5∼6 years (12.29%). When 209 cases (40.12%), including double infection were analyzed, there were 88 cases (16.89%) of norovirus-GII infection, 26 cases (4.99%) of Campylobacter spp. infection, 18 cases (3.45%) of rotavirus infection, 18 cases (3.45%) of Clostridium difficile Toxin B infection, 17 cases (3.26%) of adenovirus infection, 16 cases (3.07%) of Clostridium perfringens infection, 11 cases (2.11%) of astrovirus infection, 5 cases (0.96%) of Salmonella spp. infection, 3 cases (0.58%) of norovirus-GI, Yersinia spp. and Aeromonas spp. infections, and 1 case (0.19%) of verocytotoxin-producing E. coli infection. Based on a seasonal separation of early childhood, norovirus-GI and norovirus-GII mainly prevailed during the winter, when the temperature is low. Typical enteritis with an increased prevalence of rotavirus during the spring. Astrovirus prevailed between the months of April and October, when norovirus-GI, norovirus-GII, and rotavirus did not prevail. With increasing age, acute diarrheal disease was not only induced by a virus, but also by bacteria. Although a test for virus is an effective method when trying to identify the cause during early childhood by multiplex-PCR, it would be desirable to undergo tests for both virus and bacteria concurrently as age increases.

다발성 골수종 환자에서 kaposi`s sarcoma-associated herpesvirus(KSHV) 감염의 임상적 의의

김찬규(Chan Kyu Kim),홍대식(Dae Sik Hong),박성규(Sung Kyu Park),이규택(Gyu Taeg Lee),원종호(Jong Ho Won),백승호(Seung Ho Baick),이동화(Dong Wha Lee),박희숙(Hee Sook Park) 대한내과학회 2000 대한내과학회지 Vol.58 No.2

N/A Background : Kaposi's sarcoma-associated herpesvirus (KSHV) been shown to be associated with human diseases including Kaposi's sarcoma, pleural effusion lymphoma, multicentric Castleman's disease. The IL-6 may both stimulate myeloma growth and prevent apoptosis of malignant plasma cells. Interestingly, viral IL-6(vIL-6), homolog to human interleukin-6(IL-6) in KSHV genome retains biologic activity. Thus, oncogenic role of the KSHV has been proposed as a pathogenesis of the multiple myeloma. We used ISH to determine the frequency of patients with multiple myeloma and plasmacytosis associated with KSHV-infected BM cells in fresh core biopsies and to determine the correlation between KSHV infection and clinical characteristics. Methods : Bone marrow(BM) biopsy samples from 16 cases of multiple myeloma, 2 cases of monoclonal gammopathy of undetermined significance(MGUS) were obtained from the pathology division of Soon Chun Hyang University Hospital, Seoul, Korea. Biopsy sample of Kaposi's sarcoma for positive control and BM biopsy samples of myelodysplastic syndrome(MDS) and malignant lymphoma for negative control were obtained. Bitinylated probe to KSHV were prepared with the following sequences: 5' to 3' TGCAGCAGCTGTTGGTGTACCACATATCT. and in situ hybridization (ISH) was performed. Results : Among the 18 patients. Two patients were MGUS and among 16 patients with multiple myeloma, 1 in stage IB disease, 1 stage IIB disease, 8 stage IIIA disease, 4 stage IIIB diseases and 2 in variant of multiple myeloma, extramedullary plasmacytoma. Strong positive signal was detected in nuclei and cytoplasm of the malignant cells of biopsy sample from 1 cases of Kaposi's sarcoma by ISH(positive control). Signal was not detected in BM biopsy samples of 7 cases from MDS and malignant lymphoma(negative control). Among 16 patients with multiple myeloma, 15 demonstrated viral positive cells and 2 cases with MGUS also showed viral positive cells by ISH. Signal was detected in nuclei and cytoplasm of stromal cells. Signal was strongly detected in MGUS than multiple myeloma. Positivity of the KSHV was not related with stage of the patients with multiple myeloma. One patients with multiple myeloma was studied at diagnosis and after chemotherapy. After chemotherapy KSHV was not detected. Conclusion : In MGUS and multiple myeloma, KSHV infects the stromal cells of BM rather than malignant plasma cells. On the basis of these data, we have supposed KSHV to play a role in transformation from MGUS to multiple myeloma. Particularly, due to the fact that signal of ISH was strongly detected in MGUS and was not detected in one case with multiple myeloma, it was presumed that KSHV was not major role in already advanced multiple myeloma but statistic significance was not demonstrated because of small numbers of cases. Further studies to reveal the correlation of KSHV and pathogenesis of multiple myeloma are needed.(Korean J Med 58:213-220, 2000)

재생불량성 빈혈의 병태생리에서 Fas 항원과 Apoptosis의 역할

원종호,이남수,김숙자,정희정,이규택,박성규,백승호,김성일,홍대식,박희숙,Won, Jong-Ho,Lee, Nam-Su,Kim, Sook-Ja,Cheong, Hee-Jeong,Lee, Kyu-Taeg,Park, Seung-Kyu,Baick, Seung-Ho,Kim, Sung-Il,Hong, Dae-Sik,Park, Hee-Sook 대한면역학회 2002 Immune Network Vol.2 No.1

Background: Clinical observations and laboratory studies have supported an immune basis for most acquired aplastic anemias, with the majority of patients responding to immunosuppressive therapy. Fas, a member of the tumor necrosis factor (TNF) receptor superfamily is a critical downregulator of cellular immune responses. Proinflammatory cytokines like interferon gamma (IFN-${\gamma}$) and TNF-${\alpha}$ can induce Fas expression and render hematopoietic progenitor cells susceptible to Fas-induced growth suppression and apoptosis. Methods: In order to investigate the involvement of apoptosis in the pathogenesis of aplastic anemia (AA), we measured the expression of Fas antigen and caspase-3 on bone marrow (BM) mononuclear cells (MNCs) of AA in the presence or absence of IFN-${\gamma}$, TNF-${\alpha}$, or macrophage inflammatory protein 1-${\alpha}$ (MIP-$1{\alpha}$). Results: We confirmed that AA BM MNCs were more apoptotic and highly expressed Fas antigen than normal donors. Stimulation by IFN-${\gamma}$, TNF-${\alpha}$, or MIP-$1{\alpha}$ increased Fas antigen and caspase-3 expression in AA BM MNCs than BM MNCs of normal donors. Anti-Fas monoclonal antibody enhanced IFN-${\gamma}$, TNF-${\alpha}$, or MIP$1{\alpha}$ mediated caspase-3 expression in BM MNCs of normal donors. Among these three cytokines, IFN-${\gamma}$ enhanced apoptosis most strongly via Fas-caspase-3 pathway. Conclusion: These results suggest that Fas signal pathway may play a role in the pathophysiology of aplastic anemia and negative hematopoietic regulators like IFN-${\gamma}$ can induce apoptosis of bone marrow progenitors in part by Fas induction.

위발성 위장관 악성림프종 환자에서 항암치료의 효과

김찬규,신영록,김현정,배상병,이남수,이규택,박성규,원종호,홍대식,박희숙 순천향대학교 2006 Journal of Soonchunhyang Medical Science Vol.12 No.1

Purpose: The gastrointestinal (GI) tract is the most common site of extranodal non-Hodgkin's lymphoma (NHL), which is increasing in incidence, but there is no established optimal treatment modality. Thus, this study was investigated the clinicohistologic feature, the therapeutic modalities, and the prognosis for GI-NHL, as well as the factors affecting it. Methods: We retrospectively analyzed 45 patients who had been diagnosed as having GI-NHL and had been followed up from July 1994 to February 2005 at Soonchunhyang University Hospital. The patients were divided into groups according to the site of origin and to various other features, and the survivals of the various groups were compared. The modified Ann Arbor system and WHO classification were adopted for staging and histopathologic classification, respectively. Results: GI-NHL of the stomach, small bowel, ileocecal region, and colon occurred in 28 patients (62.2%), 5 patients (11.1%), 3 patients (6.7%), and 8 patients (17.8%), respectively, In one patient, the entire gastrointestinal tract was diffusely involved. The median age of patients was b5 years (25~78 years), and male-to-female ratio was 1:1.1. Fourteen patients were in stage Ⅰ, 24 in stage Ⅱ, 4 in stage Ⅲ, and 3 in stage Ⅳ. Surgical resection was performed in 19 patients, and combination chemotherapy was performed in 43 patients. Surgical resection only was performed in 4 patients, Chemotherapy only was performed in 26 patients. The expected overall 5 year survival of 45 patients was 39.6%, and there was a significant survival difference between the stages, but between sites of origin (p=0.842). The most important factors influencing the survival was the stage and other factors were not significant. Conclusion: The stomach was the most common site of GI-NHL. Most GI-NHL were localized Stage was the most important prognostic factor. However, Prospective randomized studies are needed to approve the therapeutic modality.

비혈연간 동종 골수이식술 후 발생한 혈전성 미세혈관병증 1례

이국경,배상병,김주성,이준혁,이유경,이규택,박성규,원종호,백승호,진소영,홍대식,박희숙 대한조혈모세포이식학회 2000 대한조혈모세포이식학회지 Vol.5 No.2

혈전성 미세혈관병증은 골수이식 후 발생하는 중한 합병증 중의 하나이다. 저자들은 22세의 급성 림프구성 백혈병(ALL, L3)을 진단 후 비혈연간 동종 골수이식을 받은 환자에서 발생한 혈전성 미세혈관병증을 경험하였다. 그는 비혈연간 동종골수이식술을 받았으며 이식의 전처치로 busulfan, cyclophosphamide, 전신방사선조사를 받았고 이식편대숙주반응을 막기 위하여 CsA와 methotrexate을 투여받았다. 이식 후 28일째 말초혈액도말검사상 Burr cell, schistocyte, tear drop cell이 관찰되었으며, 이식 후 31일째 혈변소견이 보여 S결장 내시경상 장관 이식편대 숙주질환 소견이 보였고 혈청검사상 거대세포바이러스가 양성소견을 보여 steroid와 gancyclovir를 투여받았고 2주 후에 장관 이식편대 숙주반응이 호전되었다. 이식 후 68일째 임상적으로 혈전성 미세혈관병증이 발생되었고 Cr 수치가 증가되어 TPE와 혈액투석을 격일로 시행하였으나 악화되었다. 이식 후 82일째 다발성 출혈로 사망하였다. Thrombotic microangiopathy (TMA) is one of the serious complications of bone marrow transplantation (BMT). We experienced a 22-year-old male with acute lymphoblastic leukemia (ALL, L3) who developed post BMT-TMA. He received pretransplant conditioning chemotherapy with busulfan, cyclophosphamide, and total body irradiation (TBI). He received cyclosporine (CsA) and methotrexate (MTX) for graft versus host disease (GVHD) prophylaxis. Peripheral blood smear revealed burr cells, schistocytes and tear drop cells on post BMT day 28. Hematochezia was developed on post BMT day 31. Biopsy specimen of sigmoid colon revealed grade III to IV acute GVHD and cytomegalovirus (CMV) culture was positive in blood on post BMT day 43. He received steroid pulse therapy and gancyclovir so that improved after 2 weeks. TMA was developed clinically on post BMT day 68 and creatine (Cr) was increased. In spite of undergoing plasma exchange with fresh frozen plasma and hemodialysis every other day, TMA progressed. He died due to multiple hemorrhage on post BMT day 86.

고용량 항암요법 및 자가 조혈모세포 이식을 시행받은 유방암 환자에서 미세진행암 검색을 위한 Cytokeratin-19 검사의 의의

정재화,정희정,이규택,박성규,원종호,백승호,서원석,홍대식,박희숙,이민혁 대한조혈모세포이식학회 2000 대한조혈모세포이식학회지 Vol.5 No.2

배경:1978년 악성 림프종에서 처음으로 자가골수이식이 성공적으로 수행된 후 자가골수이식이 급속도로 증가하여 최근에는 동종골수이식보다 더 많이 시술되고 있다. 이러한 자가 조혈모세포이식은 유방암 환자에서 최근 그 빈도가 증가하고 있는 추세이나 자가이식의 잠재적인 한계인 골수나 말초 조혈모세포 수집으로부터 얻은 오염된 조혈모세포의 재주입의 가능성 여지가 있다는 것이 문제점으로 지적되고 있다. 최근 자가 조혈모세포이식에 따른 잔존 세포암에 대한 중요성이 증가되어져 왔고 이는 잔존 암세포를 발견할 수 있는 분자적 기술의 발전에 기인하고 있다. 유방암 환자에 있어 정상 혈액 단핵구내 유방암 세포 발견을 위해 cytokeratin-19 (CK-19) mRNA와 epithelial growth factor receptor는 민감도와 특이도가 뛰어난 표지자로 알려져 있다. 최근에 세포 특이 단백질에 대한 mRNA를 RT-PCR에 의존한 검사로서 10^(7)개의 정상세포중 최소 1개의 tumor cell을 발견할 수 있게 되었다. 본 연구에서는 autograft 내의 유방암 세포의 발견을 위한 RT-PCR 방법의 민감도를 평가하고 CK-19에 대한 RT-PCR 검사 양성인 말초혈액 내 암세포의 존재와 임상양상 결과를 비교 분석하였다. 방법:1996년 9월부터 1999년 12월까지 순천향대학병원에서 입원하여 고용량 항암요법 및 자가 조혈모세포 이식을 시행받은 환자들을 대상으로 하였다. 이들 총 13명의 환자중에서 10개 이상의 림프절 양성을 보이는 고위험군 유방암 환자가 6명이었고 재발성 유방암 환자가 7명이었다. 대상환자들에게 2번의 FAC 요법이후 mobilization으로 doxorubicin, paclitaxel 및 GM-CSF를 투여하였고 전처치로는 thio-tepa, carboplatin, mitoxantrone을 사용하였다. Mobilization 이후 채취한 조혈모세포의 일부를 이용하여 CK-19에 대한 RT-PCR을 시행하였다. 결과:RT-PCR의 민감도는 유방암 세포주인 MCF-7을 이용하였을때 정상 골수세포 100만개 중 1개의 MCF-7 세포가 존재하여도 RNA가 증폭되는 결과를 얻음으로써 감수성은 1:10^(6)의 비를 보였다. CK- 19에 대한 RT-PCR 결과는 13명의 환자 중 2명에서 CK-19 양성 소견을 보임으로써 15.4%의 양성률을 보였다. 2명의 환자는 모두 재발된 유방암 환자군에 속했으며 이중 1명은 1번에서만 CK-19 양성 소견을 보였고 나머지 한명은 2번 모두 apheresis 내에 CK-19 mRNA를 포함하고 있었다. 또한 전체 생존율을 비교하였을 때 고위험군에서 2년 생존율이 52.6%, 재발된 군에서 19.1%가 관찰되었으나 PCR 양성인 2명의 재발환자는 각각 4개월과 16개월째 사망하였다. 결론:고위험군과 재발된 유방암 환자에서 고용량 항암요법 및 자가조혈모세포 이식은 비교적 안전하며 잠재적 이득이 있는 치료법이다. 유방암 환자의 말초혈액 조혈모세포 내에 미세 잔존암 여부를 확인 위해 시행한 cytokeratin-19에 대한 RT-PCR은 특이도와 민감도가 뛰어난 검사이며 말초혈액 조혈모세포 내 잔존 암세포의 존재는 무병 생존률이나 전체 생존율을 감소시키는 경향을 보여 향후 유방암에서 조혈모세포이식을 시행할 때 잔존 암세포의 검색을 시행할 필요가 있을 것으로 생각되며 채집된 조혈모세포 내에 오염되어 있는 암세포의 제거 방법의 개발이 필요할 것으로 생각된다. Background:Breast cancer is a chemosensitive tumor and dose intensification might improve the disease-free survival and cure rates in high risk and relapsed breast cancer patients with poor prognosis. Autologous peripheral blood stem cells (PBSCs) provide accelerated hematopoietic recovery after myeloablative and myelosuppressive chemotherapy. The appreciation of the importance of minimal residual disease has been associated with a dramatic increase in reports describing application of technology to detect minimal disease, especially molecular techniques based primarily on the polymerase chain reaction. Cytokeratin-19 mRNA and epithelial growth factor receptor mRNA were shown to be highly specific and sensitive markers for the detection of breast cancer cells within normal peripheral blood mononuclear cells. The purpose of this study is to establish a sensitivity of RT-PCR assays for the detection of breast cancer cells within autografts and to evaluate the correlation between clinical outcome and the presense of tumor cells in PBSCs that positively reacted to RT-PCR assays. Methods:A cytokeratin-19 (CK-19) specific-nested RT-PCR assay was developed andoptimized by using limited dilutions of the MCF-7 breast cancer line mixed with normal bone marrow cells. The optimized assay was then used to examine PBPCs samples obtained from 13 patients with high risk and relapsed breast cancer. Results: In the sensitivity, CK-19 expressing tumor cells were detected in the mixture of 10 MCF-7 cells in 10^(7) normal bone marrow cells. Two patients of 13 (15.4%) scored positive to CK-19 in PBSCs with breast cancer, indicating minimal residual disease. Two patients with relapsed breast cancer with positive of CK-19 were died at 4 and 16 months after PBSCT. Conclusion: RT-PCR of cytokeratin-19 assay is a sensitive, specific and rapid method for the detection of minimal residual cancer cells within PBSCs in breast cancer patients. The presence of minimal residual tumor in PBSCs was trending toward short median disease-free survival and overall survival and this assay can be used for detection of cancer cells in harvested PBSCs in breast cancer patients with high dose therapy with autologous PBSCT.

Systemic Sclerosis Following Autologous Peripheral Blood Stem Cell Transplantation for Non-Hodgkin's Lymphoma

Kim, Hyun-Jung,Kim, Sung-Han,Kim, Chan-Kyu,Roh, Hak-Jae,Jin, So-Young,Jung, Jin-Tae,Lee, Nam-Su,Lee, Kyu-Taeg,Park, Sung-Kyu,Won, Jong-Ho,Hong, Dae-Sik,Park, Hee-Sook 대한조혈모세포이식학회 2003 대한조혈모세포이식학회지 Vol.8 No.2

Autoimmune disease can develop after stem cell transplantation (SCT), and of these systemic sclerosis usually result from allogeneic SCT. However, no case has been reported of systemic sclerosis developing after autologous SCT. We experienced a case of systemic sclerosis in a patient 3 years after autologous peripheral blood SCT for recurrent non-Hodgkin's lymphoma (NHL). The pathogenesis of the condition is unclear, though possible mechanisms may involve an imbalance of immune regulation induced by thymic damage, hormonal factors, a genetic predisposition, or environmental factors.