말초혈액 조혈모세포 채집 예측인자: 단일기관 분석

장민중,조덕,신명근,양동욱 대한수혈학회 2014 大韓輸血學會誌 Vol.25 No.3

Background:Peripheral blood stem cell (PBSC) transplantation is a curative treatment in various hematologic malignancies and some solid cancers. Effective mobilization and collection of PBSC is essential for successful PBSC transplantation. The aim of this study was to investigate the useful factors for predicting PBSC collection using multivariate analysis. Methods:We retrospectively reviewed the medical records of 170 allogeneic and 389 autologous donors at Chonnam National University Hwasun Hospital between 2005 and 2012. Donor groups were divided into three groups (failure group, suboptimal group, and optimal group) according to the total CD34+ yield. Donors were compared regarding age, sex, body weight, disease, complete blood count, hematopoietic progenitor cell (HPC) parameter of automated cell counter, process volume, number of leukapheresis procedures, prior mobilization history, type of vascular access and instrument. Results:In allogeneic PBSC collections (n=170), the collection failure group showed lower baseline (premobilization) white blood cell (WBC) (P=0.004) and HPC (P<0.001) than the optimal group. In autologous PBSC collections (n=389), the collection failure group showed lower baseline HPC and more frequent prior mobilization history (P<0.001) than the suboptimal and optimal group. In multivariate analysis, older age, lower number of leukapheresis procedures, and prior mobilization history were risk factors associated with mobilization failure. Conclusion:Our data suggest that baseline WBC and HPC would be useful for predicting poor mobilizer in allogeneic PBSC collection, whereas baseline HPC would be useful in autologous PBSC collection. Conventional chemotherapy and G-CSF based remobilization would not be helpful to proven poor mobilizer in previous mobilization.

In Vitro Fluconazole and Voriconazole Susceptibilities of Candida Bloodstream Isolates in Korea: Use of the CLSI and EUCAST Epidemiological Cutoff Values

장민중,신종희,이위교,김미나,이경원,이혜수,이미경,장철훈,장희창,송은송,김수현,신명근,서순팔,양동욱 대한진단검사의학회 2013 Annals of Laboratory Medicine Vol.33 No.3

Background: At present, the clinical breakpoints (CBPs) of both fluconazole and voriconazole are available only for 3 common Candida species in the Clinical and Laboratory Standards Institute (CLSI) and the European Committee on Antimicrobial Susceptibility Testing (EUCAST) methods. Epidemiological cutoff values (ECVs) were recently applied to both methods to detect the emergence of acquired resistance (i.e., non-wild-type isolates)among 5 common Candida species. Methods: We performed a nationwide study to determine the fluconazole and voriconazole susceptibility of Candida bloodstream isolates (BSIs) using both the CLSI and EUCAST methods. A total of 423 BSIs of 5 Candida species were collected from 8 hospitals. The azole susceptibilities were assessed on the basis of the species-specific CBPs and ECVs. Results: Of the 341 BSIs of 3 common Candida species (i.e., C. albicans, C. tropicalis,and C. parapsilosis), 0.3% and 0.9%, 0.0% and 1.5% of isolates were categorized as fluconazole and voriconazole resistant according to the CLSI and EUCAST CBPs, respectively. Of 423 total BSIs, 1.4% and 2.6% had fluconazole minimum inhibitory concentrations (MICs) exceeding the ECVs according to the CLSI and EUCAST, respectively; 1.0% and 2.1% had voriconazole MICs exceeding the ECVs according to the CLSI and EUCAST, respectively. Categorical agreement between the methods using ECVs was 98.3% for fluconazole and 98.3% for voriconazole. Conclusions: The EUCAST and CLSI methods using ECVs provide highly concordant results. Moreover, non-wild-type isolates with possibly acquired azole resistance were rare among the BSIs of 5 common Candida species in Korea.

Autoantibodies with Mimicking Specificity Detected by the Dilution Technique in Patients with Warm Autoantibodies

장민중,조덕,박경운,Mark Harris Yazer,신명근,신종희,서순팔,양동욱 대한진단검사의학회 2013 Annals of Laboratory Medicine Vol.33 No.5

Background: The aim of this study was to investigate the frequency of autoantibodies with mimicking specificity by using the dilution technique, to assess the usefulness of the combination of the dilution technique and red blood cell (RBC) phenotyping, and to establish a pre-transfusion testing algorithm in patients with warm autoantibodies. Methods: Serum samples from 71 patients with warm autoantibodies were tested using the dilution technique. Among them, 25 samples were adsorbed with allogeneic ZZAP (a combination of dithiothreitol and enzyme) or polyethylene glycol (PEG) and their RBC phenotypes were determined. Thirty-nine patients were transfused with our pre-transfusion testing algorithm using a combination of dilution technique and RBC phenotyping. Results: Autoantibodies with mimicking specificity were detected by the dilution technique in 26.8% (19/71) of the patients and most of them were directed against Rh system antigens. The agreement of the results obtained with the dilution technique in combination with RBC phenotyping and those from ZZAP or PEG adsorption was 100% (18/18) in patients who have autoantibodies with mimicking specificity and/or alloantibodies. No clinical symptoms indicating severe acute or delayed hemolytic transfusion reactions were reported in the 39 patients transfused with our pre-transfusion testing algorithm. Conclusions: Autoantibodies with mimicking specificity detected by the dilution technique in patients with warm autoantibodies are relatively frequent, can be discriminated from alloantibodies by employing a combination of dilution technique and RBC phenotyping, and might not appear to cause severe acute or delayed hemolytic transfusion reactions.

창유리용 필름이 창호의 단열성능에 미치는 영향

장민선(Min-Seon Jang),강재식(Jae-Sik Kang),최경석(Gyeong-Seok Choi),최현중(Hyun-Jung Choi),김유민(Yumin Kim) 대한설비공학회 2013 대한설비공학회 학술발표대회논문집 Vol.2013 No.6

공공 건축물의 에너지 성능 개선을 위한 외피시스템 리모델링 적용방안

최현중(Hyun-Jung Choi),김유민(Yumin Kim),장민선(Min-Sun Jang),강재식(Jae-Sik Kang),최경석(Gyeong-Seok Choi) 대한설비공학회 2013 대한설비공학회 학술발표대회논문집 Vol.2013 No.6

Cis-AB 가계에서 발견된 새로운 O02 변이형 - 한 가족 내 3 증례 보고

원은정,장민중,신동준,신명근,양동욱,조덕 대한수혈학회 2010 大韓輸血學會誌 Vol.21 No.2

In preoperative ABO typing, we observed an ABO discrepancy in a 61 year-old patient with A2B3 phenotype. Standard serologic tests for the ABO blood group phenotypes and ABO gene direct sequencing for the exons 6 and 7 were done as part of the patient’s family study. We found that the patient’s genotype was identical to cis-AB01/O02, except for a 703 G>A polymorphism at exon 7. An allele-separation by cloning and subsequent sequencing of the heterozygote was carried out. We identified a novel O02 variant allele characterized by a 703 G>A polymorphism in the patient and in her 2 daughters. (Korean J Blood Transfus 2010;21:148-153) 저자들은 61세 여자 환자의 ABO 불일치를 해결하는 과정에서 혈청학적으로 cis-AB형을 의심하고 실시한 ABO 유전자의 액손 6과 7 분석에서액손 7의 703 G>A만을 제외하고는 cis-AB01/O02와 일치한 소견을 보여, 가계도 조사, 가족들ABO 혈액형의 혈청학적 분석, 유전형 분석 그리고 환자에 대해서는 추가로 클로닝을 실시하여cis-AB01 대립유전자와 O02 대립유전자를 분리한 후 다시 직접염기서열분석을 통해 703 G>A 변이가 새로운 O02 변이형에서 기원하였음을 증명하여 보고하는 바이다.

온난자가항체 존재시 혈청희석법으로 증명한 Anti-Fyb 특이성을 보인 자가항체 - 증례보고 -

정은선,장민중,임채승,양동욱,조덕 대한수혈학회 2010 大韓輸血學會誌 Vol.21 No.1

Several approaches have been introduced to detect allo-antibodies in the presence of warm auto-antibodies, and these methods include warm autoadsorption, cysteine-activated papain and dithiothreitol (ZZAP), and polyethylene glycol (PEG) and dilution of the patient’s serum. Among them, the dilution technique is a simple and rapid method. During pretransfusion testing of a 33 year-old systemic lupus erythematosus (SLE) patient with warm auto-antibodies, antibody identification was done by the dilution technique with using serum diluted 1-in-8. The patient demonstrated an anti-Fyb pattern of reactivity in his sera. Contrary to our expectations, the phenotype of the erythrocytes was Fy(a+/b+) and the genotype, as assessed by performing PCR-restriction fragment length polymorphism (RFLP), was FY*A/FY*B. These results suggest that the antibody is an autoantibody showing anti-Fyb specificities. An antibody identification test using undiluted serum showed the same result when 40 days had passed. We report here on a case with auto-anti-Fyb proven by the dilution method in the presence of warm autoantibodies. (Korean J Blood Transfus 2010;21:74-79) 온난자가항체가 존재할 경우에 동종항체를 검출하는 방법으로 온난자가흡착법, ZZAP, PEG, 그리고 환자의 혈청 희석법이 소개되었다. 이중 혈청희석법은 간편하고 신속한 검사법이다. 저자들은 33세 SLE환자의 수혈전 검사로 의뢰된 불규칙항체 선별 및 동정 검사에서 ‘혈청희석법’으로 환자의 혈청을 8배 희석하여 자가항체의 역가를 의미있게 감소시키고, anti-Fyb 특이성을 보이는 항체를 동정하였다. 그런데 예상과 달리, 환자의 Duffy 혈액형 검사에서 Fy(a+/b+)를, PCR- RFLP를 이용한 유전형 검사에서 FY*A/FY*B를 보여, 항체는 동종항체가 아닌 anti-Fyb 특이성을 보이는 또 다른 자가항체임을 확인할 수 있었다. 환자는 외래 추적 40일 후에 희석하지 않은 혈청으로 실시한 검사에서도 anti-Fyb 특이성이 다시 확인되었기에 보고하는 바이다.

새로운 국산 V-Tube 진공채혈관과 BD Vacutainer의 비교평가

원은정,신명근,장민중,조덕,기승정,김수현,신종희,원용관,양동욱,서순팔 대한진단검사의학회 2013 Laboratory Medicine Online Vol.3 No.3

Background: Vacuum tubes are widely used in clinical laboratories for routine tests. We compared a newly developed V-tube (AB Medical, Korea)and BD tubes (BD, USA) in common clinical assays, i.e., hematological, chemical, and immunological tests. Methods: In total, 100 volunteers comprising 79 patients and 21 healthy volunteers were recruited and peripheral blood samples were collected with 2 brands of EDTA tubes and serum-separating tubes (SSTs). EDTA-tube samples were evaluated using 16 routine hematological tests. The SST samples were analyzed for 32 routine chemical parameters and 3 thyroid hormones. The results were statistically analyzed using the paired ttest and Bland-Altman plot. In addition, the stability of each analyte in 2 brands of vacutainers was evaluated. The results of the hematological tests at t=0 hr were compared with those at t=72±2 hr, and the results of the chemical parameters and thyroid hormones at t=0 hr were compared with those at t=72±2 hr and t=168±2 hr for each tube. Results: Paired t-test analysis revealed that the test results of 16 routine hematological parameters, 32 routine chemical parameters, and 3 thyroid hormones showed clinically allowable differences between the 2 brands of vacuum tubes (t =0 hr). The results obtained when using V-tubes showed a statistically significant correlation with those obtained when using BD tubes. The stability of each analyte was similar in both vacuum tubes. Except for 10 parameters (white blood cell count, mean corpuscular volume, basophils [ %], mean corpuscular hemoglobin concentration,monocytes [ %], phospholipid, sodium, potassium, chloride, and free T4), all parameters showed significant but clinically allowable differences with regard to storage duration. Conclusions: The newly developed V-tube vacutainers provide a suitable alternative to BD tubes in common clinical laboratories. 배경: 진공채혈관은 임상검사에서 널리 쓰이고 있다. 본 연구자들은 최근 개발된 V-tube와 BD Vacutainer 진공채혈관을 일반혈액검사, 일반화학검사 및 면역학적 검사를 대상으로 비교 평가하였다. 방법: 총 100명의 성인(건강인 21명, 환자 79명)을 대상으로 본 연구를 시행하였다. 말초혈액을 ethylene diamine tetra acetic acid(EDTA) tube와 serum separator tube (SST) 각각에 채혈하여 두 진공채혈관을 비교하였다. EDTA tube로는 일반혈액검사 16종을, SST로는 일반화학 32종 및 갑상선 기능검사 3종에 대하여 검사를 시행하였다. 두 진공채혈관의 검사 결과는 쌍체비교법과 Bland-Altmanplot을 이용해 비교하였다. 안정성 검사를 위해서 두 진공채혈관 각각의 채혈당일 검사결과를 기준으로 하여 일반혈액검사는 채혈 후3일째의 검사치의 변화를, 일반화학 및 갑상선기능검사는 채혈 후3일, 7일째의 검사치의 변화를 검사 항목별로 분석하였다. 결과: 당일 검체로 시행한 두 진공채혈관의 측정값 간 차이는 일반혈액검사 16종, 일반화학검사 32종, 갑상선기능검사 3종 모두에서높은 일치도를 보였고, 임상적으로 허용 가능한 범위 내에 속하였다. 또한 두 진공채혈관의 측정값 간에는 통계학적으로 유의한 강한 양의 상관관계를 보였다. 보관 기간에 따른 검사 안정성 비교에서 두 진공채혈관 모두 유사한 정도의 안정성을 보였다. 당일 검체와 3일째 검체의 비교 시 WBC, MCV, Basophil은 두 채혈관 모두에서 임상적으로 유의한 차이를 보였고, BD 진공채혈관에서는MCHC, Monocyte에서 임상적으로 유의한 차이를 보였다. 일반화학검사와 갑상선 기능검사의 경우 의 경우 당일 검체와 보관 검체(3일, 7일)와의 결과 비교 시 두 가지 진공채혈관 모두에서 대부분의 항목의 결과가 통계적으로 유의한 차이가 있음을 알 수 있었다. 하지만 PL, Na, K, Cl, free T4 항목을 제외한 대부분은 임상적으로허용할 만한 오차 범위 내의 결과를 보였다. 결론: 임상검사실에서 흔히 시행하는 검사들에 대해 새로 개발된V-tube를 BD Vacutainer 진공채혈관과 비교 평가한 결과 국산 Vtube진공채혈관이 수입제품을 대체할 수 있을 정도의 품질과 성능을 보유한 것으로 평가되었다.

단클론감마병증에서 면역글로불린 중쇄 및 경쇄 복합체 직접 측정의 유용성: 모세관 전기영동과의 수행능 비교

강민구,신명근,김진각,장민중,이오진,김혜란,조덕,김수현,기승정,신종희,서순팔,양동욱 대한진단검사의학회 2014 Laboratory Medicine Online Vol.4 No.1

Background: Determination of monoclonal gammopathy through conventional protein electrophoresis is sometimes difficult because of the presence of large proteins such as haptoglobin and transferrin, which may obscure the results. Ambiguity in an electrophoresis band can give rise to confusion or difficulty in interpretation. The heavy chain/light chain assay (HLC assay) using Hevylite antibody (The Binding Site, UK) has recently been developed for the accurate measurement of monoclonal proteins. We compared the immunotyping (IT) profiles to the immunoglobulin (Ig) heavy/light chain measurements obtained using the HLC assay and observed the ratios between intact Ig kappa and lambda. Methods: We collected 35 and 28 sera from patients with suspicious and definitive monoclonal protein, respectively. Then we performed serum protein electrophoresis (SPEP) and IT by Capillarys2 (Sebia, USA). Monoclonal protein production was investigated using Freelite antibody (The Binding Site) and specific Ig(G, A)κ and Ig(G, A)λ Hevylite antibodies. The results were analyzed using PASW 18.0 for Windows (IBM, USA). Results: Direct measurement of Ig heavy/light chains showed discordant IT results for 12 (34.2%) of 35 patients’ sera with suspicious SPEP pattern and identical IT results for 28 patients’ sera with definitive monoclonal peak in the SPEP results. Overall, the results of the HLC assay and IT showed good agreement (κ=0.718, P=0.000 by cross-tabulation Gamma, Kappa analysis). Conclusions:The results of direct measurement of serum Ig heavy chain/light chain pairs were comparable to those of IT and were helpful for determination of monoclonality in the case of ambiguous electrophoresis results. Measurement of the heavy chain/light chain pair ratio also allowed precise quantification of the monoclonal Igs with ambiguous electrophoresis patterns and identification or discrimination of clonality. 배경: 전통적인 단백 전기영동에 의한 단클론감마병증의 확인은 합토글로빈, 트랜스페린과 같은 큰 농도를 가진 단백에 의해 가려지므로 어려울 수 있다. 전기영동 밴드에서의 이와 같은 불확실성으로 인해 혼란스럽거나 해석에 어려움이 있을 수 있다. 최근 M단백의 정확한 측정을 위해서 Hevylite 항체(The Binding Site, UK)를 사용하는 Heavy chain/light chain assay (HLC assay)가 개발되었기에, 면역 전기영동에 의한 M단백의 형별판정과 면역글로불린의 중쇄 및 경쇄 쌍의 직접 측정을 하여 얻은 완전 면역글로불린의 카파 및 람다 비율에 의한 형별판정의 결과를 비교하였다. 방법: Capillarys2(Sebia, Norcross, USA)에 의한 혈청 단백 전기영동과 면역 전기영동에서 의심스러운 소견을 보인 35검체와 확실한 단클론성을 보인 28검체를 각각 모았다. M단백의 생산은 Freelite 항체(The Binding Site, UK)와 특이적인 Ig(G, A)κ and Ig(G, A)λ Hevylite 항체(The Binding Site)에 의해서도 측정되었다. 검사결과는 PASW 18.0에 의해서 분석되었다. 결과: 면역글로불린 중쇄/경쇄 쌍의 직접 측정은 의심스러운 전기영동 소견을 보인 35검체 중에서는 12검체(34.2%)에서 면역전기영동에 의한 M단백의 형별판정과 일치하지 않았고, 혈청단백 전기영동에서 확실한 단클론성 피크를 보였던 28검체에서는 면역전기영동에 의한 M단백의 형별판정과 모두 일치한 소견을 보였다. 전반적으로, HLC assay와 면역 전기영동검사 결과는 M단백의 형별판정에 있어서 좋은 일치율을 보였다(κ=0.718, P=0.000, 일치도 분석). 결론: 혈청 면역글로불린 중쇄/경쇄 쌍의 직접측정은 면역전기영동과 동등한 유용성을 보이며, 전기영동에서 애매한 단클론성을 보이는 경우 형별판정을 하는 데 도움이 될 수 있다. 또한 애매한 전기영동 소견을 보이는 M단백의 경우, 정량을 하여 형별판정의 확인 및 감별을 하는 데 유용하다고 하겠다.

혈소판성분채집세트내 잔여혈액에서 자연살생세포의 체외 증폭

신동준,조덕,장윤영,박지윤,김주선,김상기,이제중,장민중,신명근,양동욱 대한수혈학회 2012 大韓輸血學會誌 Vol.23 No.1

Background:Natural killer cells expanded from human peripheral blood (PB) have been used in cancer immunotherapy research. Although most research teams have access to human PB, it is necessary to find a source of blood that can be easily obtained. We have tested the possibility of using blood retained in a disposable platelet apheresis set as an alternative source, with special interest in expansion of NK cells for use in cancer immunotherapy research. Methods:For expansion of NK cells, peripheral blood mononuclear cells (PBMCs) were isolated from an MCS+ platelet apheresis kit (Haemonetics, Braintree, USA) and PB from the same donor (n=7) and co-cultured with 100-Gy gamma ray-irradiated K562 cells expressing the 4-1BB ligand and membrane-bound IL-15 for three weeks in RPMI1640 medium in the presence of IL-2 and IL-15. Cytotoxicity was measured using WST-1 at 1:1, 2:1, and 4:1 effector-to-target (E:T) ratios for a period of four hours. Results:Mean rate of expansion of NK cells was 1,097-fold and their purity was 94.4% from blood retained in a disposable platelet apheresis set; mean rate of expansion of NK cells was 953-fold and their purity was 92.0% from PB after a period of three weeks. No differences in cytotoxicity against K562, 697, Raji, and RPMI8226 were observed between NK cells expanded from two blood sources. Conclusion:Blood retained in a disposable platelet apheresis set is a useful and convenient source for expansion of NK cells for use in cancer immunotherapy research.