• 대한의생명과학회지
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• 제28권4호
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• pp.298-306
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• 2022

Pathological tissue fixation using formalin has been widely used for histological samples in many hospitals and institutions. In general, formalin fixatives were either manufactured in laboratories or purchased commercially because of the risks and environmental concerns of handling organic compounds. In this study, the efficacy of three kinds of commercially purchased and one laboratory-made formalin fixative was compared in the PCR-based molecular diagnosis using the extracted DNA from formalin-fixed paraffin-embedded (FFPE) tissues. The quality of extracted DNA from FFPE tonsil tissues with four kinds of formalin solutions was evaluated, and PCR for beta-globin gene and microsatellite instabilities (MSI) tests for pentaplex panel markers were performed using the extracted DNA. There was no difference in PCR and MSI tests as molecular diagnoses regardless of the types of formalin used in this study. However, the total amount and average length of double-stranded DNA extracted from FFPE tonsil tissue showed significant differences according to the type of formalin fixative. Optimized formalin fixatives and methods for DNA extraction might be sophisticated to extract good quality DNA from the small size of specific tissue samples. Further studies are needed to select the most effective formalin fixative for histology and molecular pathology using human FFPE tissues.

• IMMUNE NETWORK
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• 제21권4호
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• pp.31.1-31.16
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• 2021

Gastric cancer (GC) is the fourth most common cause of cancer-related death globally. The classification of advanced GC (AGC) according to molecular features has recently led to effective personalized cancer therapy for some patients. Specifically, AGC patients whose tumor cells express high levels of human epidermal growth factor receptor 2 (HER2) can now benefit from trastuzumab, a humanized monoclonal Ab that targets HER2. However, patients with HER2^negative AGC receive limited clinical benefit from this treatment. To identify potential immune therapeutic targets in HER2^negative AGC, we obtained 40 fresh AGC specimens immediately after surgical resections and subjected the CD45⁺ immune cells in the tumor microenvironment to multi-channel/multi-panel flow cytometry analysis. Here, we report that HER2 negativity associated with reduced overall survival (OS) and greater tumor infiltration with neutrophils and non-classical monocytes. The potential pro-tumoral activities of these cell types were confirmed by the fact that high expression of neutrophil or non-classical monocyte signature genes in the gastrointestinal tumors in The Cancer Genome Atlas, Genotype-Tissue Expression and Gene Expression Omnibus databases associated with worse OS on Kaplan-Meir plots relative to tumors with low expression of these signature genes. Moreover, advanced stage disease in the AGCs of our patients associated with greater tumor frequencies of neutrophils and non-classical monocytes than early stage disease. Thus, our study suggests that these 2 myeloid populations may serve as novel therapeutic targets for HER2^negative AGC.

• 대한유전성대사질환학회지
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• 제20권2호
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• pp.55-62
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• 2020

HT-1은 FAH의 돌연변이에 의한 FAH의 결함으로 타이로신 대사의 중간산물인 FAA, SA가 간세포, 신세뇨관 세포에 축적되어 증상을 일으키는 대사이상 질환의 일종이다. 생후 6개월 내에 증상이 발현되는 급성형의 경우 간기능 저하에 따른 간부전, 응고장애, Fanconi 증후군, FAA 축적에 의한 델타-아미노레부릭산 탈수효소의 기능 저하에 의한 포르피린증 유사 증상이 발생한다. 증상이 생후 12개월 이후 발현되는 만성형의 경우 간경화, 간세포암종, 저인산혈증 구루병, 인지기능 저하와 같은 증상이 발생할 수 있다. 본 증례는 생후 20개월 경 간비장비대, 내반슬, 대상산증으로 대사이상질환 감별을 위하여 시행한 탠덤매스 분석에서 특이 소견이 보고되지 않았으나, 57개월에 시행한 복부 초음파 검사에서 간경화, 소변 유기산 분석에서 SA 상승, NGS에서 FAH 돌연변이 c.107T>C (p.Ile 36Thr), c.614T>C (pPhe205Ser)가 확인되어 HT-1이 진단된 증례로, 환아는 현재까지 NTBC 및 단백제한식이를 유지하며 특이 합병증 없이 외래 추적관찰 중이다. HT-1은 혈장 타이로신 농도가 증가하지 않는 경우도 있으며, 만성형의 경우 임상양상이 생후 12개월 이후에 나타나므로 탠덤매스 검사에서 음성 소견이 보고되어도 임상적으로 의심되는 경우 소변 유기산 분석과 같은 추가적 검사가 필요하다. 또한 기존의 탠덤매스 분석을 통하여는 HT-1이 진단되지 않는 경우가 있을 수 있어, 신생아 선별검사에서 혈장 타이로신 농도뿐 아니라 SA 농도도 추가로 확인하는 것의 비용 편익에 대한 추가적 연구가 필요하다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권16호
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• pp.6871-6876
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• 2015

Background: The meningeal hemangiopericytoma (MHPC) is a vascular tumor arising from pericytes. Most intracranial MHPCs resemble meningiomas (MNGs) in their clinical presentation and histological features and may therefore be misdiagnosed, despite important differences in prognosis. Materials and Methods: We report 8 cases of MHPC and 5 cases of MNG collected from 2007 to 2011 from the Neuro-Surgery and Histopathology departments. All 13 samples were re reviewed by two independent pathologists and investigated by immunohistochemistry (IHC) using mesenchymal, epithelial and neuro-glial markers. Additionally, we screened all tumors for a large panel of chromosomal alterations using multiplex ligation probe amplification (MLPA). Presence of the NAB2-STAT6 fusion gene was inferred by immunohistochemical staining for STAT6. Results: Compared with MNG, MHPCs showed strong VIM (100% of cases), CD99 (62%), bcl-2 (87%), and p16 (75%) staining but only focal positivity with EMA (33%) and NSE (37%). The p21 antibody was positive in 62% of MHPC and less than 1% in all MNGs. MLPA data did not distinguish HPC from MNG, with PTEN loss and ERBB2 gain found in both. By contrast, STAT6 nuclear staining was observed in 3 MHPC cases and was absent from MNG. Conclusions: MNG and MHPC comprise a spectrum of tumors that cannot be easily differentiated based on histopathology. The presence of STAT6 nuclear positivity may however be a useful diagnostic marker.

• Journal of Genetic Medicine
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• 제16권2호
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• pp.67-70
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• 2019

Limb-girdle muscular dystrophy (LGMD) is a group of muscular dystrophies that has extremely heterogeneous clinical features and genetic background. The caveolin-3 gene (CAV3) is one of the causative genes. LGMD appears as a clinical continuum, from isolated skeletal muscle involvement to long QT syndrome. Here we report two patients without apparent muscle weakness in a family with CAV3 mutation. A 7-month-old Korean boy visited our muscle clinic because of an incidental finding of elevated serum creatine kinase (CK) concentration (680 IU/L, reference range, 20-270 IU/L) without clinical symptoms. The patient was born after an uneventful pregnancy and showed normal developmental milestones. He developed pseudohypertrophy of his calf muscle during the follow-up. We obtained a muscle biopsy at age 14 months, which showed size variations and degenerating/regenerating myofibers with endomysial fibrosis and immunohistochemical evidence of normal dystrophin. Under the impression of LGMD, we performed target panel sequencing and identified a heterozygous in-frame mutation of CAV3, c.307_312delGTGGTG (p.Val103_Val104del). Immunohistochemical staining of muscle indicated complete loss of caveolin-3 compared with normal control muscle, which supported the variant's pathogenicity. We performed segregation analysis and found that the patient's mother had the same variant with elevated serum CK level (972 IU/L). We report on autosomal dominant familial caveolinopathy caused by a pathogenic variant in CAV3, which was asymptomatic until the fourth decade. This case highlights the utility of next generation sequencing in the diagnosis of muscular dystrophies and the additive role of muscle biopsy to confirm the variants.

• Asian Pacific Journal of Cancer Prevention
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• 제15권14호
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• pp.5539-5544
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• 2014

Background: To evaluate the location of tumor relapse and imaging modality for detection according to the breast cancer subtype: luminal A, luminal B, HER2 positive luminal B, nonluminal HER2 positive, and triple negative. Materials and Methods: A total of 1244 patients with breast cancer with known estrogen receptor (ER), progesterone receptor (PR), Ki-67 and human epidermal growth factor receptor 2 (HER2), who underwent breast surgery from 2009 to 2012 were analyzed. Patients were classified into the following categories: luminal A (n=458), luminal B (n=241), HER2 positive luminal B (n=227), nonluminal HER2 positive (n=145) and triple negative (n=173). A total of 105 cases of relapse were detected in 102 patients: locoregional recurrence (n=46), recurrence in the contralateral breast (n=28) and distant metastasis (n=31). Comparison of proportions was used to determine the difference between subtypes. Results: Relapse rates by subtypes are as follows: luminal A 23 of 458 (5.02%), luminal B 19 of 241(7.88%), HER2 positive luminal B 15 of 227 (6.61%), nonluminal HER2 postive 19 of 145 (13.10%) and triple negative 29 of 173(16.76%). Luminal A tumors had the lowest rate of recurrence and had significantly lower recurrence rate in comparison with nonluminal HER2 postive (p=0.0017) and triple negative subtypes (p<0.0001). Compared with all other subtypes except nonluminal HER2 positive, triple negative tumors had the highest rate of tumor recurrence (p<0.01). Triple negatives were most likely to develop contralateral recurrence against all subtypes (p<0.05). Detection rate of locoregional and contralateral tumor recurrence were 28.3% on mammography (n=17/60). Conclusions: Luminal A tumors are associated with a low risk of recurrence while triple negative lesions have a high risk. In case of triple negative tumors, the contralateral breast has much more recurrence as compared with all other subtype. In terms of detection rates, breast USG was the best modality for detecting tumor recurrence, compared with other modalities (p<0.05). Subtyping of breast tumors using a molecular gene expression panel can identify patients who have increased risk of recurrence and allow prediction of locations of tumor recurrence for each subtype.

• 한국식품저장유통학회지
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• 제22권1호
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• pp.108-118
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• 2015

꾸지뽕 열매의 부가가치를 증대하기 위해 꾸지뽕 발효식초를 제조하고 그의 발효조건을 확립하였다. 전통발효식품으로부터 초산내성, 초산 고생산능, ethanol 내성 및 아황산 내성이 우수한 49종의 초산 균주를 분리하였고, 16S rRNA 유전자 염기서열의 해독 결과, Acetobacter indonesiensis, A. cerevisiae, A. orientalis, A. tropicalis, A. fabarum, A. pasteurianus 및 A. syzygii으로 동정되었다. 이들 중 GRAS 균주인 A. pasteurianus SCMA5와 SCMA6를 발효 균주로 최종 선정하였다. 최적 발효는 꾸지뽕 열매 함량이 40%(v/v)인 즙액과 5%(v/v) ethanol을 첨가하여 $25^{\circ}C$에서 72시간 발효가 가장 적절하였다. 관능평가 결과, SCMA06 균주를 적용한 발효액의 선호도가 SCMA05 균주를 적용한 발효액보다 높았다. SCMA06 균주를 사용한 발효식초에서 항산화 능력을 측정하는 DPPH 라디컬 소거활성의 경우 대조구에 비해 $53.02{\pm}0.78%$이상 높게 나타났고, 항당뇨 능력을 측정하는 AGI활성은 발효 72시간에 $91.40{\pm}2.43%$ 저해능을 보여 시판중인 acarbose보다 활성이 높았다. 이번 연구는 꾸지뽕 열매를 활용한 발효식초 제조를 위한 산업화 연구에 기여할 수 있을 것이다.

• 한국수정란이식학회지
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• 제18권3호
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• pp.243-248
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• 2003

1. 생쥐 고환으로부터 얻은 세포를 배양하여 군집을 형성하는 것을 관찰할 수 있었으며, AP, SSEA-1, -3, -4과 Integrin $\alpha$6, $\beta$1 및 Oct4의 발현을 확인하였다. 2. 생쥐 생식줄기세포를 3-5일정도 배양하게 되면, 여러 층으로 이루어진 군집을 이루게 되는데 이는 생쥐 배아줄기세포나 배아생식줄기세포의 형태와 같은 것이었다. 3. 생쥐 생식줄기세포를 체외에서 효과적으로 분리, 배양할 수 있는 조건을 확립하였다.

• Clinical and Experimental Pediatrics
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• 제61권12호
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• pp.403-406
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• 2018

Floating-Harbor syndrome is a rare autosomal dominant genetic disorder associated with SRCAP mutation. To date, approximately 50 cases of Floating-Harbor syndrome have been reported, but none have been reported in Korea yet. Floating-Harbor syndrome is characterized by delayed bony maturation, unique facial features, and language impairment. Here, we present a 6-year-old boy with a triangular face, deep-set protruding eyes, low-set ears, wide nose with narrow nasal bridge, short philtrum, long thin lips, clinodactyly, and developmental delay that was transferred to our pediatric clinic for genetic evaluation. He showed progressive delay in the area of language and cognition-adaption as he grew. He had previously undergone chromosomal analysis at another hospital due to his language delay, but his karyotype was normal. We performed targeted exome sequencing, considering several syndromes with similar phenotypes. Library preparation was performed with the TruSight One sequencing panel, which enriches the sample for about 4,800 genes of clinical relevance. Massively parallel sequencing was conducted with NextSeq. An identified variant was confirmed by Sanger sequencing of the patient and his parents. Finally, the patient was confirmed as the first Korean case of Floating-Harbor syndrome with a novel SRCAP (Snf2 related CREBBP activator protein) mutation (c.7732dupT, p.Ser2578Phefs*6), resulting in early termination of the protein; it was not found in either of his healthy parents or a control population. To our knowledge, this is the first study to describe a boy with Floating-Harbor syndrome with a novel SRCAP mutation diagnosed by targeted exome sequencing in Korea.

• 대한유전성대사질환학회지
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• 제22권1호
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• pp.28-36
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• 2022

목적: 혈장 아미노산(PAA) 및 소변 유기산(UOA) 분석에서 비정상적인 대사 산물의 검출은 리 증후군과 같은 임상 미토콘드리아 질환을 진단하는 데 사용되었다. 본 연구에서는 PAA 및 UOA 분석의 진단적 가치와 유효성을 검토하였다. 방법: 이 논문은 2003년에서 2018년 사이에 단일 3차 진료 센터에서 진단된 리 증후군 환자에 대상으로 후향적 연구로 진행되었다. 전체 미토콘드리아 시퀀싱 및 핵 DNA 관련 미토콘드리아 유전자 패널 분석을 통해 미토콘드리아 DNA (mtDNA) 돌연변이 관련 리 증후군에 대해 19명의 환자가 양성이었고 57명의 환자는 음성인 것으로 밝혀졌다. 그 이후에 PAA 및 UOA 분석 결과를 비교하였다. 결과: 두 그룹 간의 PAA 및 UOA 분석 결과를 비교한 결과, mtDNA 돌연변이 양성 Leigh 증후군과 mtDNA 돌연변이 음성 Leigh 증후군 그룹 간에 비정상적인 대사 산물은 뚜렷한 차이를 보이지 않았다. 결론: PAA 및 UOA 분석은 리 증후군을 진단하거나 mtDNA 돌연변이 관련 리 증후군을 선별하기 위한 부적절한 검사 방법이다. 그러나 UOA 분석은 여전히 리 증후군에 대한 적합한 선별 검사일 수 있다.