• 한국환경보건학회지
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• 제27권2호
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• pp.82-91
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• 2001

This study was carried out to elucidate the protective effect of zinc chloride(ZnCl$_2$) and its mechanism against the immuno-cytotoxicity of methylmercury chloide($CH_3$HgCl). This study was observed in the culture of EMT-6 cells which are originated from mammary adenocarcinoma of Balb/c mouse. Cytotoxicity of metals was measured by cell viability and NO$_2$$^{[-10]}$ , and mitochondrial function was evaluated by adenosine triphosohate (ATP) production. $CH_3$HgCl significantly decreased the sythesis of nitric oxide(NO), ATP and glutathione(GSH) in a dose-dependent manner. ZnCl$_2$ significantly increased the synthesis of GSH in a dose-dependent manner, but synthesis of NO and ATP were not changed. The immuno-cytotoxicity of $CH_3$HgCl was not fully protected when combined addition of ZnCl$_2$, whereas ZnCl$_2$ prior to addition of $CH_3$HgCl completly protected the Hg-induced immuno-cytotoxicity. Similarly, intracellular accumulation of mercury significantly decreased by ZnCl$_2$. Degree of diminution of intracellular mercury was larger in ZnCl$_2$ prior to addition of $CH_3$HgCl than in combined addition of ZnCl$_2$ and $CH_3$HgCl.. Dithiothreitol(DTT) or buthionine sulfoximine(BSO) addition at 50$\mu$M or less, which was not toxic to the cells, did not affect synthesis of NO and ATP. DTT increased intracellular GSH level and DTT pretreatment protected toxicity induced by $CH_3$HgCl as shown complete recover in the NO and ATP values. BSO decreased intracellular GSH level and BSO pretreatment exaggerated toxicity induced by $CH_3$HgCl as shown synergistic reduction in the NO and ATP values. These results indicated that the protective effects of zinc against immuno-cytotoxicity of methylmercury associated with increasing cellular level of GSH. Increased intracellular GSH transports methylmercury to out of cells. In accordance with intracellular level of mercury decreased, immuno-cytotoxicity of methylmercury decreased. These result also suggest that the protective mechanism of zinc against the mercury toxicity would be exerted in the immune system in vivo.

• 생명과학회지
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• 제13권4호
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• pp.529-534
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• 2003

홍화씨가 골절, 골다공증 및 골형성부전증 등의 골 질환 치료에 효과가 있다는 것이 알려져 민간요법으로 사용되고 있으나 홍화씨의 어떤 성분이 이러한 작용을 하는지에 관해서는 거의 밝혀져 있지 않다. 본 연구에서는 흥화씨의 sterol 조성을 분석하고 폐경기 여성들의 골다공증 치료에 사용되고 있는 합성 estrogen 대체물질로써 그 기능이 확인된 phytoestrogen 활성을 측정하였다. Cholesterol에서 성호르몬인 estrogen이 합성되는 것처럼 식물 sterol 도 phytoestrogen 합성을 위한 전구체 작용을 하는 것으로 알려져 있다. 홍화씨에 존재하는 식물성 sterol은 stigmast-5-en-3-ol (3$\beta$, 24S)-form인 ${\gamma}$-sitosterol (clionasterol)로 확인되었으며 홍화에 존재하는 식물성 sterol의 함량은 총지질의 약 4%를 함유하였다. 홍화씨의 sterol 분석은 GC-MS와 Wiley MS spectrum library를 사용하여 분석하였으며 홍화씨에 존재하는 중요한 4-desmethyl sterols 에는 sitosterol 만이 확인되었다. 홍화씨의 식물성 estrogen 효과는 $\beta$-estradiol ($10^{-8}$ M)의 활성을 1로 했을 때 홍화씨의 에탄을 층(5 mg/ml) 및 메탄올 층(5 mg/ml)에서 각각 0.37과 0.43으로 estrogen 유사효과를 나타내었다(ANOVA p<0.001). 홍화씨에 포함된 estrogen에 의한 전사활성 촉진은 MCF7/pDS-CAT-ERE119-Ad2MLP system을 사용하여 estrogen수용체와 reporter gene 발현을 이용하는 방법으로 세포에 직접 처리함으로써 phytoestrogen 활성을 측정할 수 있었다.

• 한국식품영양과학회지
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• 제33권6호
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• pp.936-940
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• 2004

본 연구는 된장 메탄올 추출물의 항암효과를 검토하기 위해서 다른 콩 관련 발효식품과 원재료인 콩과 밀가루의 메탄올추출물과 비교하면서 여러 인체 암세포들의 성장 억제 실험과DNA 합성 저해 실험을 행하였다. AGS 인체 위암세포의 경우 첨가농도 200 $\mu\textrm{g}$/mL에서 콩된장과 70% 콩된장 메탄올 추출물은 각각 80%, 78%의 저해효과를 가졌으며 청국장 메탄올 추출물은 65%, 일본의 미소 메탄올 추출물은 54%의 저해효과를 보였다. Hep 3B 인체 간암세포에 대한 증식억제 효과 결과에서도 콩된장과 70% 콩된장의 경우, 첨가농도 200 $\mu\textrm{g}$/mL에서 각각 77%, 73%의 저해효과를 가지는 반면, 청국장, 일본의 미소는 각각 60%, 56%의 억제효과를 보였고 콩과 콩/밀가루의 경우 각각 56%, 40%의 저해효과를 나타내었다. HT-29 인체 대장암세포에서도 이상의 간암, 위암세포의 결과와 유사하게 증식억제효과를 보여 콩된장과 70% 된장 메탄올 추출물이 각각 86%, 87%의 저해효과를 나타내면서 다른 콩관련 발효식품들과 원재료 콩에 비해 큰 활성을 보였다. 또한 DNA 합성 저해 실험에서 AGS 인체 위암세포는 된장 메탄올 추출물 100 $\mu\textrm{g}$/mL, 200 $\mu\textrm{g}$/mL 투여시에는 각각 65%, 76%의 DNA 합성 저해 효과가 관찰되었고, 반면 콩의 메탄올 추출물의 경우 첨가농도 100 $\mu$/mL, 200 $\mu$/mL 때 각각 47%, 68%의 DNA 합성 저해 효과를 가졌으며 된장의 경우보다 낮은 저해 효과를 나타내었다. Hep 3B 인체 간암세포는 된장 메탄올 추출물 100 $\mu\textrm{g}$/mL, 200 $\mu\textrm{g}$/mL 투여시 각각 43%, 59%의 DNA합성 저해 효과를 나타내었다. 이상의 연구 결과들로부터 된장이 다른 콩 발효식품 및 원재료 콩보다 높은 암세포 성장 억제 효과와 DNA 합성 저해 효과를 가지는 것은 콩만으로 3개월 간 발효시킨 된장의 우수성과 함께 발효과정을 거치는 동안 원재료인 콩에서는 없었던 혹은 함량이 적은 성분들이 생성되거나 증가되어 항암효과를 나타내는 것으로 추정되어진다.

• 대한방사성의약품학회지
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• 제5권1호
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• pp.18-25
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• 2019

2-Nitroimidazole derivatives have been reported to accumulate in hypoxic tissue. We prepared a novel $^{99m}Tc-MAG_3$-2-nitroimidazole and evaluated the feasibility for hypoxia imaging agent. $Bz-MAG_3$-2-nitroimidazole was synthesized by direct coupling of $Bz-MAG_3$ and 2-nitroimidazole using dicyclohexylcarbodiimide. $Bz-MAG_3$-2-nitroimidazole was labeled with $^{99m}Tc$ in the presence of tartaric acid and $SnCl_2-2H_2O$ at $100^{\circ}C$ for 30 min. And the reaction mixture was purified by $C_{18}$ Sep-pak cartridge. The labeling efficiency and the radiochemical purity were checked by ITLC-SG/acetonitrile. The tumor was grown in balb/c mice for 8~13 days after the subcutaneous injection of tumor cells, CT-26 (murine colon adenocarcinoma cell). Biodistribution study and tumor autoradiography were performed in the xenografted mice after i.v injection of 74 kBq/0.1 mL and 19 MBq/0.1 mL of $^{99m}Tc-MAG_3$-2-nitroimidazole, respectively. In vivo images of $^{99m}Tc-MAG_3$-2-nitroimidazole in tumor bearing mice were obtained 1.5 hr post injection. The labeling efficiency was $45{\pm}20%$ and the radiochemical purity after purification was over 95%. Paper electrophoresis confirmed negative charge of $^{99m}Tc-MAG_3$-2-nitroimidazole. $^{99m}Tc-MAG_3$-2-nitroimidazole was very stable at room temperature and its protein binding was 53%. The $^{99m}Tc-MAG_3$-2-nitroimidazole exhibited high uptake in the liver, stomach and intestine. In biodistribution study using tumor bearing mice, the uptakes (% ID/g) of the tumor were $0.5{\pm}0.1$, $0.4{\pm}0.0$, $0.2{\pm}0.1$ and $0.1{\pm}0.1$ at 5, 15, 30 min and 4 hrs. Tumor/muscle ratio were $1.4{\pm}0.1$, $2.2{\pm}0.83$, $3.0{\pm}0.9$, and 3.7 (n=2) for 5, 15, 30 min and 4 hrs. The uptake in hypoxic area was found higher than in non-hypoxic area of tumor tissue by autoradiography. In vivo images showed the relatively faint uptake to the hypoxic tumor region. $^{99m}Tc-MAG_3$-2-nitroimidazole was successfully synthesized and found feasible for imaging hypoxia.

• Journal of Gastric Cancer
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• 제19권1호
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• pp.121-131
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• 2019

Purpose: The significance of neuroendocrine differentiation (NED) in gastric carcinoma (GC) is controversial, leading to ambiguous concepts in traditional classifications. This study aimed to determine the prognostic threshold of meaningful NED in GC and clarify its unclear features in existing classifications. Materials and Methods: Immunohistochemical staining for synaptophysin, chromogranin A, and neural cell adhesion molecule was performed for 945 GC specimens. Survival analysis was performed using the log-rank test and univariate/multivariate models with percentages of NED ($P_{NED}$) and demographic and clinicopathological parameters. Results: In total, 275 (29.1%) cases were immunoreactive to at least 1 neuroendocrine (NE) marker. GC-NED was more common in the upper third of the stomach. $P_{NED}$, and Borrmann's classification and tumor, lymph node, metastasis stages were independent prognostic factors. The cutoff $P_{NED}$ was 10%, beyond which patients had significantly worse outcomes, although the risk did not increase with higher $P_{NED}$. Tumors with ${\geq}10%$ NED tended to manifest as Borrmann type III lesion with mixed/diffuse morphology and poorer histological differentiation; the NE components in this population mainly grew in insulae/nests, which differed from the predominant growth pattern (glandular/acinar) in GC with <10% NED. Conclusions: GC with ${\geq}10%$ NED should be classified as a distinct subtype because of its worse prognosis, and more attention should be paid to the necessity of additional therapeutics for NE components.

• Tuberculosis and Respiratory Diseases
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• 제82권3호
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• pp.227-233
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• 2019

Background: Programmed death-ligand 1 (PD-L1), a transmembrane protein, binds to the programmed death-1 (PD-1) receptor, and anti-PD-1 therapy enables immune responses against tumors. This study aimed to assess clinical characteristics of PD-L1 expression using immunohistochemistry among Korean patients with lung cancer. Methods: We retrospectively reviewed the data of patients with pathologically proven lung cancer from a single institution. PD-L1 expression determined by Tumor Proportion Score (TPS) was detected using 22C3 pharmDx (Agilent Technologies) and SP263 (Ventana Medical Systems) assays. Results: From July 2016 to July 2017, 267 patients were enrolled. The main histologic type was adenocarcinoma (69.3%). Most participants were smokers (67.4%) and had clinical stage IV disease (60.7%). In total, 116 (42%) and 58 (21%) patients had TPS ${\geq}1%$ and ${\geq}50%$, respectively. The patients were significantly older in TPS ${\geq}1%$ group than in TPS <1% group ($64.83{\pm}9.38years$ vs. $61.73{\pm}10.78years$, p=0.014), not in TPS ${\geq}50%$ cutoff value ($64.69{\pm}9.39$ vs. $62.36{\pm}10.51$, p=0.178). Regarding histologic grade, higher proportions of poorly differentiated tumor were observed in the TPS ${\geq}1%$ (40.8% vs. 25.8%, p=0.020) and TPS ${\geq}50%$ groups (53.2% vs. 27.2%, p=0.004). Among 34 patients examined with 22C3 and SP263 assays, 27 had positive results in both assays, with a cutoff of TPS ${\geq}1%$ (r=0.826; 95% confidence interval, 0.736-0.916). Conclusion: PD-L1 expression, defined as TPS ${\geq}1%$, was related to older age and poorly differentiated histology. There was a similar distribution of PD-L1 expression in both 22C3 and SP263 results.

• Journal of Veterinary Science
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• 제21권2호
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• pp.26.1-26.14
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• 2020

Pancreatic ductal adenocarcinoma is a lethal cancer type that is associated with multiple gene mutations in somatic cells. Genetically engineered mouse is hardly applicable for developing a pancreatic cancer model, and the xenograft model poses a limitation in the reflection of early stage pancreatic cancer. Thus, in vivo somatic cell gene engineering with clustered regularly interspaced short palindromic repeats is drawing increasing attention for generating an animal model of pancreatic cancer. In this study, we selected Kras, Trp53, Ink4a, Smad4, and Brca2 as target genes, and applied Campylobacter jejuni Cas9 (CjCas9) and Streptococcus pyogens Cas9 (SpCas9) for developing pancreatic cancer using adeno associated virus (AAV) transduction. After confirming multifocal and diffuse transduction of AAV2, we generated SpCas9 overexpression mice, which exhibited high double-strand DNA breakage (DSB) in target genes and pancreatic intraepithelial neoplasia (PanIN) lesions with two AAV transductions; however, wild-type (WT) mice with three AAV transductions did not develop PanIN. Furthermore, small-sized Cjcas9 was applied to WT mice with two AAV system, which, in addition, developed high extensive DSB and PanIN lesions. Histological changes and expression of cancer markers such as Ki67, cytokeratin, Mucin5a, alpha smooth muscle actin in duct and islet cells were observed. In addition, the study revealed several findings such as 1) multiple DSB potential of AAV-CjCas9, 2) peri-ductal lymphocyte infiltration, 3) multi-focal cancer marker expression, and 4) requirement of > 12 months for initiation of PanIN in AAV mediated targeting. In this study, we present a useful tool for in vivo cancer modeling that would be applicable for other disease models as well.

• BMB Reports
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• 제55권12호
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• pp.609-614
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• 2022

Mutation of the gene for adenomatous polyposis coli (APC), as seen in Apc^Min/+ mice, leads to intestinal adenomas and carcinomas via stabilization of β-catenin. Transmembrane 4 L six family member 5 (TM4SF5) is involved in the development of non-alcoholic fatty liver disease, fibrosis, and cancer. However, the functional linkage between TM4SF5 and APC or β-catenin has not been investigated for pathological outcomes. After interbreeding Apc^Min/+ with TM4SF5-overexpressing transgenic (Tg^TM4SF5) mice, we explored pathological outcomes in the intestines and livers of the offspring. The intestines of 26-week-old dual-transgenic mice (Apc^Min/+:Tg^TM4SF5) had intramucosal adenocarcinomas beyond the single-crypt adenomas in Apc^Min/+ mice. Additional TM4SF5 overexpression increased the stabilization of β-catenin via reduced glycogen synthase kinase 3β (GSK3β) phosphorylation on Ser9. Additionally, the livers of the dualtransgenic mice showed distinct sinusoidal dilatation and features of hepatic portal hypertension associated with fibrosis, more than did the relatively normal livers in Apc^Min/+ mice. Interestingly, TM4SF5 overexpression in the liver was positively linked to increased GSK3β phosphorylation (opposite to that seen in the colon), β-catenin level, and extracellular matrix (ECM) protein expression, indicating fibrotic phenotypes. Consistent with these results, 78-week-old Tg^TM4SF5 mice similarly had sinusoidal dilatation, immune cell infiltration, and fibrosis. Altogether, systemic overexpression of TM4SF5 aggravates pathological abnormalities in both the colon and the liver.

• 한국간담췌외과학회지
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• 제26권1호
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• pp.91-97
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• 2022

Backgrounds/Aims: Metastatic lesions of the pancreas (PMET) account for 1%-5% of all malignant solid pancreatic lesions (SPL). In this study we evaluated the utility of endoscopic ultrasonography with fine needle aspiration (EUS-FNA) in diagnosing PMET. Methods: Patients who underwent EUS-FNA at a community referral center between 2011-2017 for SPL were identified. Clinical, radiologic, and EUS-FNA features of those with PMET were compared to those with primary solid tumors of the pancreas: pancreatic adenocarcinoma (PDAC) and neuroendocrine tumors (PNET). Results: A total of 191 patients were diagnosed with solid pancreatic malignancy using EUS-FNA: 156 PDAC, 27 PNET, and eight (4.2%) had PMET. Patients with PMET were less likely to have abdominal pain (25.0% vs. 76.3% vs. 48.2%; p < 0.01) or obstructive jaundice (37.5% vs. 58.3% vs. 0%; p < 0.01) compared to PDAC and PNET. Those with PMET were more likely to have mass lesions with/without biliary or pancreatic ductal dilatations (100% vs. 86.5% vs. 85.2%; p < 0.01) and lower CA19-9 (82.5 ± 43.21 U/mL vs. 4,639.30 ± 11,489.68 U/mL vs. 10.50 ± 10.89 U/mL; p < 0.01) compared to PDAC and PNET. Endosonographic features were similar among all groups. Seven (87.5%) patients with PMET had a personal history of malignancy prior to PMET diagnosis. The primary malignancy was renal cell carcinoma in five PMET. Conclusions: PMET are exceedingly rare, comprising less than 5% of SLP. Patients with PMET are less likely to present with symptoms and mostly identified by surveillance imaging for the primary malignancy.

• Tuberculosis and Respiratory Diseases
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• 제43권5호
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• pp.709-719
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• 1996

연구 배경: 비소세포 폐암의 근치적 치료는 수술적 절제에 주로 의존하고 있고 근치적 수술을 받은 환자의 예후는 수술당시의 병기가 중요하며 진행된 병기의 환자일수록 총 생존기간 및 무병생존기간이 현저하게 단축됨이 많은 연구자에 의해 보고되었다. 또한 근치적 수술 후의 재발은 수술부위보다는 원격 전이에 의한 재발이 많으므로 수술 후의 생존율을 향상시키기 위해서는 원격전이에 의한 재발을 억제할 수 있는 효과적인 수술 후 보조요법이 필요하다. 수술 후의 보조요법은 방사선 치료 및 항암제 투여가 있으나 각각의 치료 방법에 따른 생존율의 보고는 연구자에 따라 상이 한 결과를 보이고 있으며 국내 연구보고는 거의 없는 실정이다. 이에 연구자등은 1990년 1월부터 1995년 12월까지 연세대학교 의과대학 부속 세브란스 병원 흉부외과에서 근치적 절제술을 시행받은 282 명의 비소세포 폐암 환자를 대상으로 수술 후 보조요법의 효과 및 예후인자에 대한 분석을 시행하였다. 방법: 후향적 연구였으며 환자의 생존율은 Kaplan-Meier방법으로 분석하였다. 환자생존에 영향을 미치는 예후인자에 대한 분석은 Cox regression model에 의한 단일 및 다중 변수분석에 준하였다. 결과: 대상 환자의 조직학적 유형으로는 편평상피암이 166예(59%), 선암이 86예(30%), 편평상피선암이 11예(3.9%), 미분화 거대세포암이 19예(7.1%)이었다. 병기는 TNM분류에 따라 l기 93명, II기 58명, IIIa기 131명이었다. 수술 후 재발한 환자는 139명이었고 국소 재발이 28예(20.1%), 원격전이가 111예(79.9)이었다. 병기에 따른 5년 생존율은 I기 50.1%, II기 3 1.3%, IIIa기 24.1%였고(p<0.0001) 병기에 따른 중앙 생존 기간은 I기 55개월, II기 27개월, IIIa기 16개월로 나타났다(p<0.0001). IIIa병기의 환자중 수술 후 항암제만을 투여받거나 항암제와 방사선치료를 받은 환자는 81명이었고 이들의 중앙 생존 기간은 19개월이었으며 수술만을 받거나 수술후 방사선 치료만을 받은 환자는 50명이었는데 이들의 중앙 생존 기간은 14개월로서 양군간에 통계적으로 유의한 차이는 없었다(p=0.2982) 또한 IIIa 병기의 환자중 수술만을 받은 환자는 11명이었고 이들의 중앙무병 생존기간은 4개월이었으며 수술후 항암제 투여만을 받은 환자는 21명이었고 이들의 중앙무병 생존기간은 16개월로 양군간에 통계적으로 의미있는 생존기간의 차이를 보였다(P=0.0494) IIIa 병기중 N2기의 환자는 92명으로 5년 생존율은 25%, 중앙 생존 기간은 15개월이었다. 이들 중 수술 후 항암제 투여군은 62명으로 중앙 생존 기간은 18개월이었고 비투여군은 30명으로 이들의 중앙 생존기간은 14개월이었으며 양군간에 통계적으로 유의한 차이는 없었다(p=0.3988). N2기 환자중 수술후 방사선 치료를 받은 환자는 66명으로 이들의 중앙 생존 기간은 16개월이었고 방사선 치료를 받지 않은 환자는 26명으로 이들의 중앙 생존 기간은 14개월이었다. 양군간에 통계적으로 유의한 생존 기간의 차이는 없었다(p=0.6588). Cox Multiple Regression Model을 이용하여 수술 후 환자의 생존율에 영향을 미치는 예후 인자에 대하여 분석한 결과 환자의 연령(p=0.0093)과 병기(p<0.0001)만이 통계적으로 유의하였다. 결론: 비소세포 폐암 환자의 연령과 병기가 유의한 예후 인자이었으며, 보조 요법에 따른 생존율에 유의한 차이는 없었다. 따라서 수술 후 환자의 생존율을 향상시키기 위해서는 더욱 효과적인 보조요법의 개발이 필요할 것으로 생각된다.