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Development of Standard Operating Procedures (SOPs), Standardization, TLC and HPTLC Fingerprinting of a Polyherbal Unani Formulation

  • Naaz, Arjumand;Viquar, Uzma;Naikodi, Mohammad Abdul Rasheed;Siddiqui, Javed Inam;Zakir, Mohammad;Kazmi, Munawwar Husain;Minhajuddin, Ahmed
    • CELLMED
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    • v.11 no.4
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    • pp.21.1-21.9
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    • 2021
  • Background: Unani System of Medicine (USM) has its origin to Greece. To ensure and develop the quality, authenticity of Unani drugs, standardization on modern analytical parameter is essential requirement for drugs. Objectives: The aimed of the present study was to develop a standard profile of "Qurṣ-e-Mafasil" by systematic study through authenticated ingredients, pharmacognostic identification followed by physicochemical, TLC, HPTLC fingerprinting analysis as per standard protocol. Material and Methods: In this study three batches of "Qurṣ-e-Mafasil" QM were prepared by standard method as per UPI had been followed by organoleptic properties of formulation such as appearance, color, odor, taste. Powder Microscopy and physicochemical studies were carried out such as Uniformity of weight, Friability, Disintegration time, hardness, LOD, ash vales and extractive values in like aqueous, alcohol & hexane. Further qualitative tests such as Thin-Layer Chromatography (TLC), and High-Performance Thin Layer Chromatography (HPTLC) studies were also carried out to develop fingerprint pattern of the alcoholic solvent extract of QM. Phytochemical screening was carried out in different solvent extracts such as alcoholic, aqueous and chloroform extracts to detect the presence phytoconstituents in the formulation QM. Heavy metals, Microbial Load Contamination and pesticidal residues were also determined. Results: Qurṣ-e-Mafasil showed tablet-like appearance, light brown colour, mild pungent odour and acrid taste. Uniformity of weight (mg), friability (rpm), and hardness (kg/cm) and disintegration time was ranged between (500 to 503), (0.0340 to 0.038), (8.40 to 8.67) and (4-5 minutes) respectively for the three batches. Loss in weight on drying at 105℃ was ranged between (8.3425 to 8.7346). Extracted values were calculated in distilled water ranged between (30.9091 to 31.4358), hexane (1.1419 to 1.4281), and alcohol (3.3352 to 3.3962). The ash values recorded were ranged between (3.7336 to 3.8378), and acid insoluble ash (0.5859 to 0.6112).

The responsibility of C-terminal domain in the thermolabile haemolysin activity of Vibrio parahaemolyticus and inhibition treatments by Phellinus sp. extracts

  • Tran Thi Huyen;Ha Phuong Trang;Nguyen Thi-Ngan;Bui Dinh-Thanh;Le Pham Tan Quoc;Trinh Ngoc Nam
    • Fisheries and Aquatic Sciences
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    • v.26 no.3
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    • pp.204-215
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    • 2023
  • The thermolabile haemolysin (tlh) of Vibrio parahaemolyticus (Vptlh) from V. parahaemolyticus is a multiple-function enzyme, initially describes as a haemolytic factor activated by lecithin and phospholipase A2 enzymatic activity (Shinoda, 1991; Vazquez-Morado, 2021; Yanagase et al., 1970). Until now, the tlh structure has hypothesized including N-terminal and C-terminal domain, but what domain of the Vptlh structure does the haemolytic activity has not been refined yet. In this study, a 450-bp VpTLH nucleotide sequence of the entire Vptlh gene encoded the C-terminal domain cloned firstly to examine its responsibility in the activity of the Vptlh. The C-terminal domain fused with a 6-His-tag named the His-tag-VpC-terminal domain was expressed successfully in soluble form in the BL21 (DE3) PlysS cell. Remarkably, both expression and purification results confirmed a high agreement in the molecular weight of the His-tag-VpC-terminal domain was 47 kDa. This work showed the His-tag-VpC-terminal domain lysed the erythrocyte membranes in the blood agar and the phosphate buffered saline (0.9%) media without adding the lecithin substrate of the phospholipase enzyme. Haemolysis occurred at all tested diluted concentrations of His-tag-VpC-terminal domain (p < 0.05), providing evidence for the independent haemolytic activity of the His-tag-VpC-terminal domain. The content of 100 ㎍ of the His-tag-VpC-terminal domain brought the highest haemolytic activity of 80% compared to that in the three remaining contents. Significantly, the His-tag-VpC-terminal domain demonstrated not to involve the phospholipase activity in Luria-Bertani agar supplemented with 1% (vol/vol) egg yolk emulsion. All results proved the vital responsibility of the His-tag-VpC-terminal domain in causing the haemolytic activity without the required activation by the phospholipase enzyme. Raw extracts of Phellinus igniarus and Phellinus pipi at 10-1 mg/mL inhibited the haemolytic activity of the His-tag-VpC-terminal domain from 67.7% to 87.42%, respectively. Hence applying the His-tag-VpC-terminal domain as a simple biological material to evaluate quickly potential derivatives against the Vptlh in vivo conditions will accessible and more advantageous than using the whole of the Vptlh.

Monitoring of Groundwater quality according to groundwater use for agriculture (농업용 지하수 사용에 따른 지하수질 모니터링 평가)

  • Ha, Kyoochul;Ko, Kyung-Seok;Lee, Eunhee;Kim, Sunghyun;Park, Changhui;Kim, Gyoo-Bum
    • Proceedings of the Korea Water Resources Association Conference
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    • 2020.06a
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    • pp.30-30
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    • 2020
  • 본 연구에서는 여름철에 농업용수(벼농사용)로서 집중적으로 지하수를 사용하는 지역에서 시기별 지하수 사용에 따른 지하수 수질변화를 평가하기 위해 수행되었다. 연구지역은 충남 홍성군 양곡리와 신곡리 일부를 포함하는 면적 2.83 ㎢(283.3 ha)에 해당하는 지역이다. 연구지역 지하수 수질의 공간적 분포 및 시간적 변화 특성 평가를 위하여 2019년 2회(7월, 10월)에 걸쳐 지하수 관정(21개소)에 대하여 조사 및 분석을 수행하였다. 지하수 샘플은 현장에서 온도(T), pH, 용존산소(DO) 및 전기전도도(EC), 산화환원전위(Eh) 등을 측정하였고, 실험실에서 주요 양이온 및 미량원소(Ca, Mg, Na, K, Si, Sr), 음이온(F, Cl, Br, NO2, NO3, PO4, SO4), 알칼리도, 용존 유기탄소(DOC)와 용존 유기물(DOM) 등을 분석하였다. 지하수 수질조사 결과, 전체의 14~15개소(67~71%)가 Ca-HCO3 유형으로 분류되었으며, 다음으로는 Ca-Cl 유형이 4~5개소(19~24%)가 관찰되었다. 얕은 심도의 관정에서 상대적으로 심도가 깊은 관정보다 대부분 성분(TDS, Ca, Mg, Na, K, Cl, SO4, HCO3, DOC)에서 높은 농도를 나타내었다. 지하수의 수질자료를 이용하여 다변량통계분석법인 주성분분석(PCA: Principal Components Analysis)과 계층적 군집분석(HCA: Hierachical Cluster Anlaysis)를 수행한 결과, 초기 3개 주요 고유성분(eigenvalue)는 PC1 54.0%, PC2 14.2%, PC3 12.3%로 전체 분산의 88.3%를 설명할 수 있었다. PC1은 Ca, Mg, Na, K, Cl, SO4, DOC가 주요한 영향 인자였으며 PC2는 HCO3, NO3, DO에 영향 받음을 확인하였다. 계층적 군집분석 결과, 연구지역 지하수는 Na-Cl 유형의 C-3 관정을 제외하고는 크게 두 그룹으로 구분되어 졌다. 다변량통계분석의 결과에서도 수리지화학, 동위원소, 용존유기물 등의 특성에서 나타나는 것과 유사한 연구지역의 수질특성을 확인할 수 있었다. 연구지역은 차시기 동안 수질변화는 일부 관정을 제외하고는 유의할 만한 수준으로 관찰되지는 않았고, 지하수 사용에 따른 지하수위 회복도 빠르게 진행되고 있는 것으로 나타났다.

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Serum exosomal miR-192 serves as a potential detective biomarker for early pregnancy screening in sows

  • Ruonan Gao;Qingchun Li;Meiyu Qiu;Su Xie;Xiaomei Sun;Tao Huang
    • Animal Bioscience
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    • v.36 no.9
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    • pp.1336-1349
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    • 2023
  • Objective: The study was conducted to screen differentially expressed miRNAs in sows at early pregnancy by high-throughput sequencing and explore its mechanism of action on embryo implantation. Methods: The blood serum of pregnant and non-pregnant Landrace×Yorkshire sows were collected 14 days after artificial insemination, and exosomal miRNAs were purified for high throughput miRNA sequencing. The expression patterns of 10 differentially expressed (DE) miRNAs were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The qRT-PCR quantified the abundance of serum exosomal miR-192 in pregnant and control sows, and the diagnostic power was assessed by receiver operating characteristic (ROC) analysis. The target genes of DE miRNAs were predicted with bioinformatics software, and the functional and pathway enrichment analysis was performed on gene ontology and the Kyoto encyclopedia of genes and genomes terms. Furthermore, a luciferase reporter system was used to identify the target relation between miR-192 and integrin alpha 4 (ITGA4), a gene influencing embryo implantation in pigs. Finally, the expression levels of miRNAs and the target gene ITGA4 were analyzed by qRT-PCR, and western blot, with the proliferation of BeWo cells detected by cell counting kit-8 (CCK-8). Results: A total of 221 known miRNAs were detected in the libraries of the pregnant and non-pregnant sows, of which 55 were up-regulated and 67 were down-regulated in the pregnant individuals compared with the non-pregnant controls. From these, the expression patterns of 10 DE miRNAs were validated. The qRT-PCR analysis further confirmed a significantly higher expression of miR-192 in the serum exosomes extracted from pregnant sows, when compared to controls. The ROC analysis revealed that miR-192 provided excellent diagnostic accuracy for pregnancy (area under the ROC curve [AUC]=0.843; p>0.001). The dual-luciferase reporter assay indicated that miR-192 directly targeted ITGA4. The protein expression of ITGA4 was reduced in cells that overexpressed miR-192. Overexpression of miR-192 resulted in the decreased proliferation of BeWo cells and regulated the expression of cell cycle-related genes. Conclusion: Serum exosomal miR-192 could serve as a potential biomarker for early pregnancy in pigs. miR-192 targeted ITGA4 gene directly, and miR-192 can regulate cellular proliferation.

The Quality Characteristics of Hamburger Patties Based on Enzyme Treated Textured Soy Protein (효소처리 조직대두단백을 이용한 햄버거패티의 품질특성)

  • Kim, Sin-Ae;Ryu, Myung-Hyun;Lee, Min-Kyoung;Oh, Jong-Shin;Kim, Sun-Ok;Lee, Sook-Young
    • Journal of the Korean Society of Food Culture
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    • v.23 no.4
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    • pp.514-520
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    • 2008
  • This study addresses the development of a soy hamburger patty containing enzyme-treated textured soy protein (TSP) as a meat analogue. In order to reduce the beany flavor and enhance the texture, TSP was treated with 0.3% Flavourzyme for 5, 10, 20, or 30 min. The degree of hydrolysis and the water holding capacity of the TSP increased with increasing hydrolysis time. The oil binding capacity of the TSP also increased with increasing hydrolysis time, approaching the maximal value, 175.82%, at 30 min, whereas that of pork scored with the lowest value of 128.67%. The volume of pork was reduced to 81.5% as the result of heat treatment, whereas that of the TSP increased to 140.57%. The values of 'L', 'b', and '${\Delta}E$' differed significantly (p<0.001) with heat treatment, but the 'a' values did not differ significantly. With regard to texture, the hardness values were highest in the pork hamburger patty (PHP), and were lowest in the soy hamburger patty (SHP) containing untreated TSP. The hardness of the SHP containing TSP treated for 20 min did not differ significantly from that of the PHP. The cohesiveness and gumminess of the SHP treated for 20 min were highest, whereas those treated for 10 min were the lowest. The gumminess of the SHP treated for 20 min did not differ significantly from that of pork. The chewiness of the PHP was the highest, whereas that of the SHP treated for 5 min was the lowest. In our sensory evaluation, PHP evidenced the highest scores, followed by the SHP treated for 30 min, as color, texture, beany flavor, and overall quality all improved as the consequence of increasing enzyme treatment duration. In conclusion, it is believed that SHP has great potential as a substitute for meat, in that the flavor, texture, and beany flavor of SHP did not differ significantly from those of PHP.

Comparative Analysis of Heavy Metal Exposure Concentrations and Volatile Organic Compound Metabolites among Residents in the Affected Area According to Residential Distance from a Coal-fired Power Plant (화력발전소 영향권 주민 거주지의 이격 거리별 중금속 및 휘발성유기화합물 대사체 노출 농도 비교 분석)

  • Jee Hyun Rho;Byoung-Gwon Kim;Jung-Yeon Kwon;Hyunji Ju;Na-Young Kim;Hyoun Ju Lim;Seungho Lee;Byeng-Chul Yu;Suejin Kim;Young-Seoub Hong
    • Journal of Environmental Health Sciences
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    • v.50 no.1
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    • pp.25-35
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    • 2024
  • Background: There are concerns about the health effects of various environmental pollution exposures among residents living near coal-fired power plants (CFPP). Objectives: This study attempted to compare the concentrations of heavy metals in blood and urine and those of urinary volatile organic compound (VOC) metabolites according to the residential separation distance. Methods: Participants in the study totaled 334 people who have lived for more than 10 years in areas within 10 km of a CFPP. The separation distance was analyzed in quartiles by dividing it into Q1 (88 people), Q2 (89 people), Q3 (89 people), and Q4 (68 people). We explained the purpose of this study to the participants and collected blood and urine after obtaining signatures on a participation agreement. Results: The study participants were 102 males (30.5%) and 232 females (69.5%), with an average age of 71. The average length of residence and distance were 43.8 years and 4,800 meters. The geometric mean concentrations of Pb, Cd, and Hg in blood and As and Cd in urine were respective 1.35 ㎍/dL, 1.43 ㎍/L, 3.16 ㎍/L. They were 167.88 ㎍/g for creatinine and 1.58 ㎍/g creatinine. The metabolite concentrations of VOCs were 50.67 ㎍/g creatinine in t, t-muconic acid (t, t-MA), 10.73 ㎍/g creatinine in benzyl mercapturic acid, 317.05 ㎍/g creatinine in phenylglyoxylic acid, 123.55 ㎍/g creatinine in methylhippuric acid, and 190.82 ㎍/g creatinine in mandelic acid. The concentration of Pb in the blood and Cd and t, t-MA in the urine of residents within affected area of the CFPP showed statistically significant differences among distance groups. Conclusions: The concentration of urinary VOCs metabolites, especially t, t-MA, differed according to the distance groups of residents within the affected area of CFPP (p<0.05).

Immune Checkpoint Inhibitor with or without Radiotherapy in Melanoma Patients with Brain Metastases: A Systematic Review and Meta-Analysis

  • Pyeong Hwa Kim;Chong Hyun Suh;Ho Sung Kim;Kyung Won Kim;Dong Yeong Kim;Eudocia Q. Lee;Ayal A. Aizer;Jeffrey P. Guenette;Raymond Y. Huang
    • Korean Journal of Radiology
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    • v.22 no.4
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    • pp.584-595
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    • 2021
  • Objective: Immune checkpoint inhibitor (ICI) therapy has shown activity against melanoma brain metastases. Recently, promising results have also been reported for ICI combination therapy and ICI combined with radiotherapy. We aimed to evaluate radiologic response and adverse event rates of these therapeutic options by a systematic review and meta-analysis. Materials and Methods: A systematic literature search of Ovid-MEDLINE and EMBASE was performed up to October 12, 2019 and included studies evaluating the intracranial objective response rates (ORRs) and/or disease control rates (DCRs) of ICI with or without radiotherapy for treating melanoma brain metastases. We also evaluated safety-associated outcomes. Results: Eleven studies with 14 cohorts (3 with ICI combination therapy; 5 with ICI combined with radiotherapy; 6 with ICI monotherapy) were included. ICI combination therapy {pooled ORR, 53% (95% confidence interval [CI], 44-61%); DCR, 57% (95% CI, 49-66%)} and ICI combined with radiotherapy (pooled ORR, 42% [95% CI, 31-54%]; DCR, 85% [95% CI, 63-95%]) showed higher local efficacy compared to ICI monotherapy (pooled ORR, 15% [95% CI, 11-20%]; DCR, 26% [95% CI, 21-32%]). The grade 3 or 4 adverse event rate was significantly higher with ICI combination therapy (60%; 95% CI, 52-67%) compared to ICI monotherapy (11%; 95% CI, 8-17%) and ICI combined with radiotherapy (4%; 95% CI, 1-19%). Grade 3 or 4 central nervous system (CNS)-related adverse event rates were not different (9% in ICI combination therapy; 8% in ICI combined with radiotherapy; 5% in ICI monotherapy). Conclusion: ICI combination therapy or ICI combined with radiotherapy showed better local efficacy than ICI monotherapy for treating melanoma brain metastasis. The grade 3 or 4 adverse event rate was highest with ICI combination therapy, and the CNS-related grade 3 or 4 event rate was similar. Prospective trials will be necessary to compare the efficacy of ICI combination therapy and ICI combined with radiotherapy.

Feeding Characteristics of Chub Mackerel Scomber japonicus in the Yellow Sea of Korea in Autumn (가을철 서해에 출현하는 고등어(Scomber japonicus)의 섭식특성)

  • Hyun-Sol Park;Changsin Kim;Jae Hyeong Yang;Se Hyun Song
    • Korean Journal of Ichthyology
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    • v.35 no.4
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    • pp.278-284
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    • 2023
  • The feeding characteristics of chub mackerel, Scomber japonicus was studied using 827 specimens caught in the Yellow Sea of Korea during Autumn season from October to December 2021 and September to November 2022. The size of the specimens ranged from 21.1 to 45.1 cm in total length. S. japonicus was a carnivore that consumed mainly on fishes, such as Engraulis japonicus (%IRI=67.9). Its diet also includes small quantities of copepods, amphipods, euphausiids, cephalopods, mysids, shrimps, and crabs, etc. The estimated trophic level of S. japonicus was 3.84. The proportion of fish had increased as the body size of S. japonicus increased, whereas those of euphausiids, copepods and amphipods decreased gradually. As the body size of S. japonicus increased, the mean weight of prey per the stomach (mW/ST) tended to increase (One-way ANOVA, F=2.382, P<0.05).

A Three-Dimensional Deep Convolutional Neural Network for Automatic Segmentation and Diameter Measurement of Type B Aortic Dissection

  • Yitong Yu;Yang Gao;Jianyong Wei;Fangzhou Liao;Qianjiang Xiao;Jie Zhang;Weihua Yin;Bin Lu
    • Korean Journal of Radiology
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    • v.22 no.2
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    • pp.168-178
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    • 2021
  • Objective: To provide an automatic method for segmentation and diameter measurement of type B aortic dissection (TBAD). Materials and Methods: Aortic computed tomography angiographic images from 139 patients with TBAD were consecutively collected. We implemented a deep learning method based on a three-dimensional (3D) deep convolutional neural (CNN) network, which realizes automatic segmentation and measurement of the entire aorta (EA), true lumen (TL), and false lumen (FL). The accuracy, stability, and measurement time were compared between deep learning and manual methods. The intra- and inter-observer reproducibility of the manual method was also evaluated. Results: The mean dice coefficient scores were 0.958, 0.961, and 0.932 for EA, TL, and FL, respectively. There was a linear relationship between the reference standard and measurement by the manual and deep learning method (r = 0.964 and 0.991, respectively). The average measurement error of the deep learning method was less than that of the manual method (EA, 1.64% vs. 4.13%; TL, 2.46% vs. 11.67%; FL, 2.50% vs. 8.02%). Bland-Altman plots revealed that the deviations of the diameters between the deep learning method and the reference standard were -0.042 mm (-3.412 to 3.330 mm), -0.376 mm (-3.328 to 2.577 mm), and 0.026 mm (-3.040 to 3.092 mm) for EA, TL, and FL, respectively. For the manual method, the corresponding deviations were -0.166 mm (-1.419 to 1.086 mm), -0.050 mm (-0.970 to 1.070 mm), and -0.085 mm (-1.010 to 0.084 mm). Intra- and inter-observer differences were found in measurements with the manual method, but not with the deep learning method. The measurement time with the deep learning method was markedly shorter than with the manual method (21.7 ± 1.1 vs. 82.5 ± 16.1 minutes, p < 0.001). Conclusion: The performance of efficient segmentation and diameter measurement of TBADs based on the 3D deep CNN was both accurate and stable. This method is promising for evaluating aortic morphology automatically and alleviating the workload of radiologists in the near future.

Protein Requirement Changes According to the Treatment Application in Neurocritical Patients

  • Jungook Kim;Youngbo Shim;Yoon-Hee Choo; Hye Seon Kim; Young ran Kim; Eun Jin Ha
    • Journal of Korean Neurosurgical Society
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    • v.67 no.4
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    • pp.451-457
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    • 2024
  • Objective : Exploring protein requirements for critically ill patients has become prominent. On the other hand, considering the significant impact of coma therapy and targeted temperature management (TTM) on the brain as well as systemic metabolisms, protein requirements may plausibly be changed by treatment application. However, there is currently no research on protein requirements following the application of these treatments. Therefore, the aim of this study is to elucidate changes in patients' protein requirements during the application of TTM and coma therapy. Methods : This study is a retrospective analysis of prospectively collected data from March 2019 to May 2022. Among the patients admitted to the intensive care unit, those receiving coma therapy and TTM were included. The patient's treatment period was divided into two phases (phase 1, application and maintenance of coma therapy and TTM; phase 2, tapering and cessation of treatment). In assessing protein requirements, the urine urea nitrogen (UUN) method was employed to estimate the nitrogen balance, offering insight into protein utilization within the body. The patient's protein requirement for each phase was defined as the amount of protein required to achieve a nitrogen balance within ±5, based on the 24-hour collection of UUN. Changes in protein requirements between phases were analyzed. Results : Out of 195 patients, 107 patients with a total of 214 UUN values were included. The mean protein requirement for the entire treatment period was 1.84±0.62 g/kg/day, which is higher than the generally recommended protein supply of 1.2 g/kg/day. As the treatment was tapered, there was a statistically significant increase in the protein requirement from 1.49±0.42 to 2.18±0.60 in phase 2 (p<0.001). Conclusion : Our study revealed a total average protein requirement of 1.84±0.62 g during the treatment period, which falls within the upper range of the preexisting guidelines. Nevertheless, a notable deviation emerged when analyzing the treatment application period separately. Hence, it is recommended to incorporate considerations for the type and timing of treatment, extending beyond the current guideline, which solely accounts for the severity by disease.