• Biomolecules & Therapeutics
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• 제27권6호
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• pp.577-583
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• 2019

Human cytochrome P450 2C9 is a highly polymorphic enzyme that is required for drug and xenobiotic metabolism. Here, we studied eleven P450 2C9 genetic variants-including three novel variants F69S, L310V, and Q324X-that were clinically identified in Korean patients. P450 2C9 variant enzymes were expressed in Escherichia coli and their bicistronic membrane fractions were prepared The CO-binding spectra were obtained for nine enzyme variants, indicating P450 holoenzymes, but not for the M02 (L90P) variant. The M11 (Q324X) variant could not be expressed due to an early nonsense mutation. LC-MS/MS analysis was performed to measure the catalytic activities of the P450 2C9 variants, using diclofenac as a substrate. Steady-state kinetic analysis revealed that the catalytic efficiency of all nine P450 2C9 variants was lower than that of the wild type P450 2C9 enzyme. The M05 (R150L) and M06 (P279T) variants showed high $k_{cat}$ values; however, their $K_m$ values were also high. As the M01 (F69S), M03 (R124Q), M04 (R125H), M08 (I359L), M09 (I359T), and M10 (A477T) variants exhibited higher $K_m$ and lower $k_{cat}$ values than that of the wild type enzyme, their catalytic efficiency decreased by approximately 50-fold compared to the wild type enzyme. Furthermore, the novel variant M07 (L310V) showed lower $k_{cat}$ and $K_m$ values than the wild type enzyme, which resulted in its decreased (80%) catalytic efficiency. The X-ray crystal structure of P450 2C9 revealed the presence of mutations in the residues surrounding the substrate-binding cavity. Functional characterization of these genetic variants can help understand the pharmacogenetic outcomes.

• Korean Journal of Acupuncture
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• 제39권4호
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• pp.172-183
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• 2022

Objectives : An error was found in the recent standard by the World Health Organization (WHO) on the locations of the Upper arm Route of Lung Meridian (URLM) and its acupoints LU3 and LU4. This possible incorrect information is being taught throughout Korean medicine colleges nationwide, which follow WHO standards. Therefore, an investigation is required to sort out this discrepancy based on the evidence in historical documents. Methods : The location of WHO's URLM and LU3 and LU4 were compared with corresponding information in the classical literature. The anatomical structure mentioned in these classical documents was examined. Finally, an assessment was conducted on whether this structure is reflected in the WHO standards. Results : Classical literature prior to the early 20th century records the locations of the Lung Meridian and LU3 and LU4 of the upper arm to be in the artery on the medial aspect. This artery corresponds to the brachial artery. The location established by the WHO is on the anterolateral side of the upper arm, where no large arteries exist that can be found by haptic search or angiographically. The anterolateral side of the upper arm belongs to the Yang aspect, which does not coincide with the Yin aspect of Lung Meridian. Conclusions : The WHO's URLM and LU3-4 standards do not agree with the classical literature. The correct route must coincide with the brachial artery passing through the medial side of the humerus. The actual location of LU3-4 is on the medial aspect of the arm, just medial to the border of the biceps brachii muscle, on the brachial artery, 3-4 B-cun inferior to the anterior axillary fold.

• 한방재활의학과학회지
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• 제25권1호
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• pp.45-52
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• 2015

Objectives To check marker content for appropriate quality control of Woohwangchungsimwon sterilized to ensure microbiological safety and to observe antioxidant activity for any changes in efficacy. Methods To measure any effects of sterilization on the effective compounds, 8 ingredients of Woohwangchungsimwon were screened for any changes in marker content using HPLC-DAD. Using the colorimetric method on the microplate reader any changes in total phenolic compound and flavonoid levels were observed. Antioxidant activity was measured using the DPPH, ABTS, and FRAP. Results Of the ingredients of Woohwangchungsimwon, 8 were subject to quantitative analysis before and after sterilization. 21.6 mg and 1.93 mg of Glycyrrhizin was found in Glycyrrhiza uralensis Fischer pre and post sterilization, respectively. Decursin found in Angelica gigas Nakai increased from 0.16 mg to 0.29 mg after sterilization. Bilirubin found in Gallstone of Bostaurusvar. domesticus increased from 0.24 mg to 0.33 mg. Cinnamic acid found in Cinnamomum cassia Blume increased from 0.02 mg to 0.05 mg. Ginsenoside Rb1 found in Panax ginseng C. A. Meyer decreased from 0.02 mg to 0.14 mg. Paeoniflorin found in Paeonia lactiflora Pallas increased from 1.05 mg to 1.13 mg. Amygdalin found in Armeniacae Amarum Semen increased from 2.68 mg to 2.83 mg. L-muscone found in Musk increased from 0.63 mg to 0.76 mg. As for total phenolic compound and total flavonoid content, there was a 1.22 and 4.15-fold increase. DPPH and ABTS increased by 20.45% and 20.69%, respectively. FRAP activity was 2.78 times more active post stabilization. Conclusions This study confirmed that high temperature high pressure steam sterilization, a method used to ensure microbiological safety of Woohwangchungsimwon, does not affect marker content; in other words, does not affect quality of the Woohwangchungsimwon. It could also be seen that total phenolic compound and flavonoid content increased after sterilization. An antioxidant activity test showed that there was significantly increased activity of antioxidants.

• 대한한방내과학회지
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• 제44권4호
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• pp.751-756
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• 2023

목적: 본 연구는 임상에서 많은 빈도로 사용되고 있는 향정신성 약물이 간독성을 발생시킬 수 있음을 임상 예를 통해 보여줌으로써, 한의진료현장에서 이에 대한 경각심과 더불어 약인성간손상에 대한 최신 정보를 제공하고자 한다. 방법: 본 임상 예의 논문은 향정신성 약물을 사용한 후 약인성간독성의 의심과 진단 및 약물의 중단 후 증상과 간손상 효소의 개선과정을 자세히 제시하였다. 결과: 평소 혼합결합조직병으로 한방병원에서 수년 동안 침과 뜸 치료 등으로 잘 유지되고 있던 56세의 여성 환자가, 어느날부터 갑자기 심한 피로감과 전신적 불편편함을 호소하였다. 혈액검사를 시행한 결과, 혈청 AST, ALT가 정상 경계의 2.5-배 이상 증가하였고, 촤근 복용한 향정신성약물에 대한 RUCAM score가 9점으로 약인성간손상 진단에 부합하였다. 이 의심 약물들을 중지한 뒤에 주관적 불편함이 빠르게 개선되었으며, 혈청 간손상 효소 수치 또한 2주 안에 정상화되었다. 결론: 본 증례는 향정신성약물로 생길 수 있는 간손상의 전형적인 임상 예로서, 향정신성약물의 일반화된 상황에 비추어 한의원에서 환자의 진료 과정에서 하나의 중요한 고려사항을 암시한다.

• Journal of Microbiology and Biotechnology
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• 제18권6호
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• pp.1179-1185
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• 2008

The immune-stimulating activities of Bordetella bronchiseptica antigens containing dermonecrotoxin (BBD) loaded in chitosan microspheres (CMs) have already been reported in vitro and in vivo with a mouse alveolar macrophage cell line (RAW264.7) and mice. Therefore, this study attempted to demonstrate the successful induction of mucosal immune responses after the intranasal administration of BBD loaded in CMs (BBD-CMs) in colostrum-deprived pigs. The BBD was introduced to the CMs using an ionic gelation process involving tripolyphosphate (TPP). Colostrum-deprived pigs were then directly immunized through intranasal administration of the BBD-CMs. A challenge with a field isolate of B. bronchiseptica was performed ten days following the final immunization. The BBD-specific IgG and IgA titers, evident in the nasal wash and serum from the vaccinated pigs, increased with time (p<0.05). Following the challenge, the clinical signs of infection were about 6-fold lower in the vaccinated pigs compared with the nonvaccinated pigs. The grades for gross morphological changes in the turbinate bones from the vaccinated pigs were also significantly lower than the grades recorded for the nonvaccinated pigs (p<0.001). Therefore, the mucosal and systemic immune responses induced in the current study would seem to indicate that the intranasal administration of BBD-CMs may be an effective vaccine against atrophic rhinitis in pigs.

• 대한방사성의약품학회지
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• 제2권2호
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• pp.108-112
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• 2016

Molecular imaging with the radiolabeled RGD peptides for ${\alpha}_v{\beta}_3$ integrin has been an increasing interest for tumor diagnosis and the treatment monitoring. Recently, $^{64}Cu$-NODAGA-gluco-E[c(RGDfK)]$_2$ was developed for quantification of ${\alpha}_v{\beta}_3$ integrin and its biological properties was elucidated. To better understand the molecular process in vivo, we performed the kinetic analysis for the $^{64}Cu$-NODAGA-gluco-E[c(RGDfK)]$_2$. After preparation of a radiotracer, dynamic PET images were obtained in the U87MG xenograft mice for 60 min (n = 6). Binding potential values were estimated from the 3-tissue compartment model, reference Logan and simplified reference tissue model. In the early time frame (0-20 min), the liver, kidney, intestine, urinary bladder and tumor were visualized but these uptakes were diminished as time went by. The tumors showed a good contrast at 40 min after administration. $^{64}Cu$-NODAGA-gluco-E[c(RGDfK)]$_2$ showed the 2-fold uptake in the tumor compared with that in the muscle. The parametric maps for binding values also provide the higher tumor-to-background contrast than the static images. A binding value obtained from the 3-tissue compartment model was comparable to other modeling methods. From these results, we conclude that $^{64}Cu$-NODAGA-gluco-E[c(RGDfK)]$_2$ may be a promising PET radiotracer for the evaluation of angiogenesis.

• Archives of Plastic Surgery
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• 제42권1호
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• pp.11-19
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• 2015

Background Wound healing is an interaction of a complex signaling cascade of cellular events, including inflammation, proliferation, and maturation. $K^+$ channels modulate the mitogen-activated protein kinase (MAPK) signaling pathway. Here, we investigated whether $K^+$ channel-activated MAPK signaling directs collagen synthesis and angiogenesis in wound healing. Methods The human skin fibroblast HS27 cell line was used to examine cell viability and collagen synthesis after potassium chloride (KCl) treatment by Cell Counting Kit-8 (CCK-8) and western blotting. To investigate whether $K^+$ ion channels function upstream of MAPK signaling, thus affecting collagen synthesis and angiogenesis, we examined alteration of MAPK expression after treatment with KCl (channel inhibitor), NS1619 (channel activator), or kinase inhibitors. To research the effect of KCl on angiogenesis, angiogenesis-related proteins such as thrombospondin 1 (TSP1), anti-angiogenic factor, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), pro-angiogenic factor were assayed by western blot. Results The viability of HS27 cells was not affected by 25 mM KCl. Collagen synthesis increased dependent on time and concentration of KCl exposure. The phosphorylations of MAPK proteins such as extracellular-signal-regulated kinase (ERK) and p38 increased about 2.5-3 fold in the KCl treatment cells and were inhibited by treatment of NS1619. TSP1 expression increased by 100%, bFGF expression decreased by 40%, and there is no significant differences in the VEGF level by KCl treatment, TSP1 was inhibited by NS1619 or kinase inhibitors. Conclusions Our results suggest that KCl may function as a therapeutic agent for wound healing in the skin through MAPK signaling mediated by the $K^+$ ion channel.

• 한국의학물리학회지:의학물리
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• 제24권4호
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• pp.220-229
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• 2013

Respiratory-induced dynamic tumors render free-breathing cone-beam computed tomography (FBCBCT) images with motion artifacts complicating the task of quantifying the internal target volume (ITV). The purpose of this paper is to study the visibility of the revealed ITV when the imaging dose parameters, such as the kVp and mAs, are varied. The $Trilogy^{TM}$ linear accelerator with an On-Board Imaging ($OBI^{TM}$) system was used to acquire low-imaging-dose-mode (LIDM: 110 kVp, 20 mA, 20 ms/frame) and high-imaging-dose-mode (HIDM: 125 kVp, 80 mA, 25 ms/frame) FBCBCT images of a 3-cm diameter sphere (density=0.855 $g/cm^3$) moving in accordance to various sinusoidal breathing patterns, each with an unique inhalation-to-exhalation (I/E) ratio, amplitude, and period. In terms of image ITV contrast, there was a small overall average change of the ITV contrast when going from HIDM to LIDM of $6.5{\pm}5.1%$ for all breathing patterns. As for the ITV visible volume measurements, there was an insignificant difference between the ITV of both the LIDM- and HIDM-FBCBCT images with an average difference of $0.5{\pm}0.5%$, for all cases, despite the large difference in the imaging dose (approximately five-fold difference of ~0.8 and 4 cGy/scan). That indicates that the ITV visibility is not very sensitive to changes in imaging dose. However, both of the FBCBCT consistently underestimated the true ITV dimensions by up to 34.8% irrespective of the imaging dose mode due to significant motion artifacts, and thus, this imaging technique is not adequate to accurately visualize the ITV for image guidance. Due to the insignificant impact of imaging dose on ITV visibility, a plausible, alternative strategy would be to acquire more X-ray projections at the LIDM setting to allow 4DCBCT imaging to better define the ITV, and at the same time, maintain a reasonable imaging dose, i.e., comparable to a single HIDM-FBCBCT scan.

• Journal of Chest Surgery
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• 제52권4호
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• pp.221-226
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• 2019

Background: Accurate mediastinal lymph node staging is vital for the optimal therapy and prognostication of patients with lung cancer. This study aimed to determine the preoperative risk factors for pN2 disease, as well as its incidence and long-term outcomes, in patients with clinical N0-1 non-small cell lung cancer. Methods: We retrospectively analyzed patients who were treated surgically for primary non-small cell lung cancer from November 2005 to December 2014. Patients staged as clinical N0-1 via chest computed tomography (CT) and positron emission tomography (PET)-CT were divided into two groups (pN0-1 and pN2) and compared. Results: In a univariate analysis, the significant preoperative risk factors for pN2 included a large tumor size (p=0.083), high maximum standard uptake value on PET (p<0.001), and central location of the tumor (p<0.001). In a multivariate analysis, central location of the tumor (p<0.001) remained a significant preoperative risk factor for pN2 status. The 5-year overall survival rates were 75% and 22.9% in the pN0-1 and pN2 groups, respectively, and 50% and 78.2% in the patients with centrally located and peripherally located tumors, respectively. In a Cox proportional hazard model, central location of the tumor increased the risk of death by 3.4-fold (p<0.001). Conclusion: More invasive procedures should be considered when preoperative risk factors are identified in order to improve the efficacy of diagnostic and therapeutic plans and, consequently, the patient's prognosis.

• 대한후두음성언어의학회지
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• 제30권2호
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• pp.107-111
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• 2019

Background and Objectives Idiopathic unilateral vocal fold paralysis (IVFP) is believed to be due to inflammation and edema of the recurrent laryngeal nerve caused by viral diseases such as upper respiratory tract infections. Corticosteroid has a potent anti-inflammatory action which should minimize nerve damage. The purpose of this study was to investigate the effect of oral steroid therapy on IVFP. Materials and Method Study was performed for the IVFP patient from January 2012 to August 2017. Patient's dermography, direction and location of paralyzed vocal cords, history of hypertension, diabetes, cerebrovascular disease, and other underlying disease, smoking history, alcohol consumption and upper respiratory tract infection, and symptoms were investigated. Treatment was divided into three groups: the observation group, low-dose group, and high-dose group, and the recovery rate and time of vocal cord paralysis were analyzed in each group. Results Thirty-seven patients were enrolled in this study. There was no relationship between oral steroid use, dosage and recovery of vocal cord paralysis. Oral steroids showed a rapid recovery of vocal cord paralysis, but there was no statistically significant difference in the time of recovery of vocal palsy with or without steroids (p=0.673). In addition, there was no statistically significant difference in recovery rate between the period to start of treatment, presence of diabetes mellitus, and treatment modality, but the recovery rate was high in the group with upper respiratory tract infection history (p=0.041). Conclusion In IVFP, oral steroid therapy has no significant difference in time and extent of recovery compared to the case of spontaneous recovery.