• Title/Summary/Keyword: Therapeutic technique

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Validation of Stem-loop RT-qPCR Method on the Pharmacokinetic Analysis of siRNA Therapeutics (Stem-loop RT-qPCR 분석법을 이용한 siRNA 치료제의 생체시료 분석법 검증 및 약물 동태학적 분석)

  • Kim, Hye Jeong;Kim, Taek Min;Kim, Hong Joong;Jung, Hun Soon;Lee, Seung Ho
    • Journal of Life Science
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    • v.29 no.6
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    • pp.653-661
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    • 2019
  • The first small interfering RNA (siRNA) therapeutics have recently been approved by the Food and Drug Administration in the U.S., and the demand for a new RNA therapeutics bioanalysis method-which is essential for pharmacokinetics, including the absorption, distribution, metabolism, and excretion of siRNA therapeutics-is rapidly increasing. The stem-loop real-time qPCR (RT-qPCR) assay is a useful molecular technique for the identification and quantification of small RNA (e.g., micro RNA and siRNA) and can be applied for the bioanalysis of siRNA therapeutics. When the anti-HPV E6/E7 siRNA therapeutic was used in preclinical trials, the established stem-loop RT-qPCR assay was validated. The limit of detection was sensitive up to 10 fM and the lower limit of quantification up to 100 fM. In fact, the reliability of the established test method was further validated in three intra assays. Here, the correlation coefficient of $R^2$>0.99, the slope of -3.10 ~ -3.40, and the recovery rate within ${\pm}20%$ of the siRNA standard curve confirm its excellent robustness. Finally, the circulation profiles of siRNAs were demonstrated in rat serum, and the pharmacokinetic properties of the anti-HPV E6/E7 siRNA therapeutic were characterized using a stem-loop RT-qPCR assay. Therefore, the stemloop RT-qPCR assay enables accurate, precise, and sensitive siRNA duplex quantification and is suitable for the quantification of small RNA therapeutics using small volumes of biological samples.

A Study on Enacting the Radiologic Technologist Act for the Civil Right to Health in Korea (건강권과 방사선사법 제정에 관한 고찰)

  • Lim, Chang-Seon
    • Journal of radiological science and technology
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    • v.30 no.4
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    • pp.313-320
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    • 2007
  • There are the Medical Radiation Health and Safety Act(the Patient Radiation Health and Safety Act, the Radiologic Technologist Act), the Medical Laboratory Technologist Act, the Physical Therapy Practice Act, and the Dental Hygienist Act, etc in America. However, Korea has only one Act for a medical radiologic technologist(including radiation therapy technologist, nuclear medicine technologist), medical laboratory technologist, physical therapist, occupational therapy examiner, dental hygienist, and so on. It is the Medical Technologist Act. Therefore, the Medical Radiation Health and Safety Act for a radiologic technologist(including radiation therapy technologist, nuclear medicine technologist) has to be enacted independently in Korea. It is the purpose of this Act to provide for the appropriate certification of persons using radioactive materials, equipment emitting ionizing radiation on humans or performing medical imaging for diagnostic and therapeutic purposes. In Korea, the radiologic technologist is a "fusion technologist" who is a person other than a licensed practitioner as a radiographer, radiation therapist, nuclear medicine technologist, computed tomography technologist, magnetic resonance technologist, mammographer, sonographer, medical dosimetrist, quality management technologist, etc. This Act will have some provisions related to the definitions, reserved title, scope of practice, specialized technologist, application for licensure, radiologic technology council, renewal, continuing education, the radiation control advisory commission, etc. This Act will ensure that quality radiation therapy treatments are delivered and that quality diagnostic information is presented for interpretation, which will lead to accurate diagnosis, treatment and cure. Accurate diagnosis can be provided only when a personnel is properly educated in technique, equipment operation and radiation safety. In the end, this Act will protect the civil right to health. By regulating the personnel responsible for performing those procedures, this Act will mean improved care for patients-higher quality images, improved accuracy, and less exposure to radiation.

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A Study on the Patterns of Alternative Therapy Experienced by the Aged (노인이 경험한 대체요법의 양상에 관한 연구 1)

  • 이강이;김순이
    • Journal of Korean Academy of Nursing
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    • v.29 no.2
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    • pp.336-345
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    • 1999
  • This study looks at the various alternative therapy methods used in day to day life by elderly, over 60 years of age. The elderly have come to know and practice these methods for the following reasons it is good for the health ; it is the method used in the of fen days when there wesn't modern medicine ; it has been passed down from generations ; it can be done at home without having the need to go to the hospital ; acupuncture or poulticing can be used ; it can be done at home, which was an important factor in rural areas where hospitals are few and far between ; and 〔herbal〕 medicine could be prepared at home at no cost ; it derives from experience ; it is impossible to ignore tradition passed down through the generations. Diet control and plants (herbs) are methods most often used. as they are easy to find and can be readily used in critical situations. Other methods include oriental medicine practices of moxibustion with moxa cone, negative therapy, hand and finger acupunture, finger press method. ordinary acupunture, manual healing methods of massage. diaphoretic therapy and meditation to reach a state of calm, and qigong dirigation. The reasons for its use are as follows ; it has been used before ; it is effective ; there is some improvement after the treatment ; it is not harmful to the body ; medicine cannot be obtained and it is the only thing available ; it is not good for an old person to go to the hospital everyday. the symptoms are not serious enough to go to a hospital : and acupuncture is for these things. The means that the elderly have come to practice these methods are : it has been used since the past ; it has been told by the elders ; they have been told by friends ; it was part of their knowledge ; and they have come to know by watching their mother. Further, to regain vitality lost through old age, the elderly have relied on hot soup. a hearty meal. brewed honey water, pumpkin, or ginseng. Humans, by instinct. would rub or massage the areas that caused pain. These actions, combined with a breathing technique have been recognized in Tong-Eui-Bo-Gam(the essential of eastern medicine), the complete work of early modern medicine, are a useful means to revive chi(기). This knowledge is thought to have greatly affected our heathy lifestyle. Furthermore, though the demand for medical services would increase with age, the elderly have not always been able to tend to their needs at the hospital for reasons economic or other. Hence, these alternative therapy methods seem to have been practiced as a temporary means of relief. The excellence of our traditional therapeutic custom has not received full recognition due to the argument relating to its scientific merits. As a result, it has become vital to prove their effectiveness through scientific and other experimental means. The potency of moxibustion with moxa cone and hand and finger acupunture have been proven scientifically. but diet and herbal methods appear to be practiced as a result of customs passed down from generations. In addition, it is submitted that the effectiveness of the traditional methods of disease control and our heathy lifestyle that are easily found in the nursing field must be verified.

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LINE-1 and Alu Methylation Patterns in Lymph Node Metastases of Head and Neck Cancers

  • Kitkumthorn, Nakarin;Keelawat, Somboon;Rattanatanyong, Prakasit;Mutirangura, Apiwat
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.9
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    • pp.4469-4475
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    • 2012
  • Background: The potential use of hypomethylation of Long INterspersed Element 1 (LINE-1) and Alu elements (Alu) as a biomarker has been comprehensively assessed in several cancers, including head and neck squamous cell carcinoma (HNSCC). Failure to detect occult metastatic head and neck tumors on radical neck lymph node dissection can affect the therapeutic measures taken. Objective: The aim of this study was to investigate the LINE-1 and Alu methylation status and determine whether it can be applied for detection of occult metastatic tumors in HNSCC cases. Methods: We used the Combine Bisulfite Restriction Analysis (COBRA) technique to analyse LINE-1 and Alu methylation status. In addition to the methylation level, LINE-1 and Alu loci were classified based on the methylation statuses of two CpG dinucleotides in each allele as follows: hypermethylation ($^mC^mC$), hypomethylation ($^uC^uC$), and 2 forms of partial methylation ($^mC^uC$ and $^uC^mC$). Sixty-one lymph nodes were divided into 3 groups: 1) non-metastatic head and neck cancer (NM), 2) histologically negative for tumor cells of cases with metastatic head and neck cancer (LN), and 3) histologically positive for tumor cells (LP). Results: Alu methylation change was not significant. However, LINE-1 methylation of both LN and LP was altered, as demonstrated by the lower LINE-1 methylation levels (p<0.001), higher percentage of $^mC^uC$ (p<0.01), lower percentage of $^uC^mC$ (p<0.001) and higher percentage of $^uC^uC$ (p<0.001). Using receiver operating characteristic (ROC) curve analysis, $%^uC^mC$ and $%^mC^uC$ values revealed a high level of AUC at 0.806 and 0.716, respectively, in distinguishing LN from NM. Conclusion: The LINE-1 methylation changes in LN have the same pattern as that in LP. This epigenomic change may be due to the presence of occult metastatic tumor in LN cases.

Effects of the Dual-Task Training on Stroke Patients : A Systematic Review and Meta-analysis (이중과제 훈련이 뇌졸중 환자에게 미치는 영향 : 체계적 고찰 및 메타분석)

  • Won, Kyung-A;Lim, Seung-Ju;Park, Hae Yean;Park, Ji-Hyuk
    • Therapeutic Science for Rehabilitation
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    • v.9 no.2
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    • pp.7-25
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    • 2020
  • Objective : The purpose of this study was to analyze the effects of dual-task training on stroke patients. Methods : We searched the databases such as NDSL, RISS, PubMed, CoChrane and EMBASE for publications in the past decade. Finally, 10 papers were selected. Qualitative assessment was performed according to the traditional single-layer evidence model, and meta-analysis was performed using the Comprehensive Meta Analysis 3.0 program. Results : The quality level of each of the 10 selected papers all correspond to I and II in the traditional single-layer evidence model. The motor tasks that constitute dual-task training comprised walking or balancing tasks in 7 articles and the motor tasks related to upper extremity were selected in 3 studies. The effect sizes for ADL function and Cognitive function were 0.65 and 0.64 (medium size effect) respectively. Moreover, the effect sizes of Lower extremity and Upper extremity motor function were 0.34 and 0.22 (small size effect) respectively. The effect size of ADL function and Cognitive function were statistically significant p<0.05). Conclusion : This study confirmed that dual-ask training can be a useful intervention technique for recovering a stroke patient's ability to perform daily activities and cognitive functions. This could be used as a helpful data when selecting appropriate intervention for stroke patients in the clinical setting.

Microarray-Based Gene Expression Profiling to Elucidate the Effectiveness of Woowhangchongshim-won on ICH Model in Rats (Microarray 분석법 활용을 통한 뇌출혈 흰쥐에서의 우황청심원 효능 평가)

  • Kim, Hyung-Woo;Cho, Su-Jin;Kim, Bu-Yeo;Jeong, Byeong-Han;Bong, Sung-Jeon;Kim, Yong-Seong;Lee, Jang-Sik;Kwon, Jeong-Nam;Kim, Young-Kyun;Cho, Su-In
    • The Korea Journal of Herbology
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    • v.22 no.4
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    • pp.253-260
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    • 2007
  • Objectives : Intracerebral hemorrhage (ICH) is characterized by breakdown of blood vessels within the brain parenchyma. Fundamental therapeutic strategies for ICH, particularly those aimed at neuroprotection, have to be established. So in this experiment, the effects of Woowhangchongshim-won, a traditional prescription formula for treating Cerebral Apoplexy in Asian countries, were investigated. Methods : After intraperitoneal injection of chloralhydrate, rats were placed in a stereotaxic frame. ICH was induced by injection of 1 U collagenase type IV and drug was administered orally for 10 days. The molecular profile of cerebral hemorrhage in rat brain tissue was measured using micro array technique to identify up- or down- regulated genes in brain tissue. These genes induced by brain damage were mainly concerned with general metabolic process such as primary metabolic process, cellular metabolic process, macromolecule metabolic process, and biosynthetic process. Results : The number of genes increased in control and not-changed in experiment was 374, and decreased in control and not-changed in experiment was 527. We are concerned with genes that can be recovered by treatment with medicine, it is especially interesting to above types of genes. Conclusions : Upon medicine treatment to the rat having cerebral hemorrhage, expressions of some genes were restored to normal level. Further analysis using protein interaction database identified some key molecules that can be used for elucidation of therapeutical mechanism of medicine in future.

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THE EXPRESSION OF TGF-$\beta$1, IGF-I, BFGF IN DISTRACTION OSTEOGENESIS ACCORDING TO DIFFERENT DISTRACTION RATES IN RABBIT'S MANDIBLE (가토 하악골에서 신연 골형성술시 신연속도에 따른 TGF-$\beta$1, IGF-I, bFGF의 발현)

  • Shin, Sun-A;Jee, Yu-Jin;Song, Hyun-Chul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.3
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    • pp.205-217
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    • 2005
  • Distraction osteogenesis is a technique of lengthening bone including soft tissue by gradual separation of surgically divided bone surfaces. Although the biomechanical, histological, and ultrastructural changes associated with distraction osteogenesis have been widely described, the molecular mechanisms governing the formation of new bone in distracted bone segments remain largely unclear. However, such information has significant clinical implications because it may enable targeted therapeutic manipulations designed to accelerate osseous regeneration. The purpose of this study was to evaluate the expression of TGF-$\beta$1, IGF-I and bFGF in distraction osteogenesis according to different distraction rates in a rabbit's mandible. When twenty-four adult rabbits underwent open osteotomy between the premolar and mental foramen, an external bilateral distraction device was applied. Latency was allowed for five days before distraction. Three different distraction rates were 0.7 mm/day (A, n=8), 1.4 mm/day (B, n=8) and 2.4 mm/day (C, n=8). The distraction device was activated with the same distraction rhythms of twice a day until 4.9 mm (A & B group) and 8.4 mm (C group) length gains was achieved. The animals were sacrificed at postoperative 3, 7, 14 and 28 days. The bony specimens were stained with H&E for histologic examination, and RT-PCR analysis was done for the identification of the expression of TGF-$\beta$1, IGF-I and bFGF. The results obtained from this study were as follows : The 0.7 mm/day and 1.4 mm/day distraction rate groups were shown to improve regenerative bone formation on radiographic and histologic examination. Also, TGF-$\beta$1, IGF-I and bFGF expression increased in the 0.7 mm/day and 1.4 mm/day distraction rate groups. But the 2.4 mm/day distraction rate group specimen was different with adjacent normal bone and hardly expressed of growth factors. These findings suggest that improved new bone formation in the 0.7 mm/day and 1.4 mm/day distraction rates is associated with enhanced expression of TGF-$\beta$1, IGF-I and bFGF by mechanical tension stress. Additionally, the 0.7 mm/day and 1.4 mm/day distraction rate groups were significantly different from the 2.4 mm/day distraction rate group in the expression of growth factors. According to the above results, it seems possible to apply a distraction rate of up to 1.4 mm/day a day in rabbit's mandible. And further studies are needed to evaluate growth factors of TGF-$\beta$1 and IGF-I, which are excellent in expression.

Construction of In Vivo Fluorescent Imaging of Echinococcus granulosus in a Mouse Model

  • Wang, Sibo;Yang, Tao;Zhang, Xuyong;Xia, Jie;Guo, Jun;Wang, Xiaoyi;Hou, Jixue;Zhang, Hongwei;Chen, Xueling;Wu, Xiangwei
    • Parasites, Hosts and Diseases
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    • v.54 no.3
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    • pp.291-299
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    • 2016
  • Human hydatid disease (cystic echinococcosis, CE) is a chronic parasitic infection caused by the larval stage of the cestode Echinococcus granulosus. As the disease mainly affects the liver, approximately 70% of all identified CE cases are detected in this organ. Optical molecular imaging (OMI), a noninvasive imaging technique, has never been used in vivo with the specific molecular markers of CE. Thus, we aimed to construct an in vivo fluorescent imaging mouse model of CE to locate and quantify the presence of the parasites within the liver noninvasively. Drug-treated protoscolices were monitored after marking by JC-1 dye in in vitro and in vivo studies. This work describes for the first time the successful construction of an in vivo model of E. granulosus in a small living experimental animal to achieve dynamic monitoring and observation of multiple time points of the infection course. Using this model, we quantified and analyzed labeled protoscolices based on the intensities of their red and green fluorescence. Interestingly, the ratio of red to green fluorescence intensity not only revealed the location of protoscolices but also determined the viability of the parasites in vivo and in vivo tests. The noninvasive imaging model proposed in this work will be further studied for long-term detection and observation and may potentially be widely utilized in susceptibility testing and therapeutic effect evaluation.

Functional Expression of P2Y Receptors in WERI-Rb1 Retinoblastoma Cells

  • Kim, Na-Hyun;Park, Kyu-Sang;Sohn, Joon-Hyung;Yeh, Byung-Il;Ko, Chang-Mann;Kong, In-Deok
    • The Korean Journal of Physiology and Pharmacology
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    • v.15 no.1
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    • pp.61-66
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    • 2011
  • P2Y receptors are metabotropic G-protein-coupled receptors, which are involved in many important biologic functions in the central nervous system including retina. Subtypes of P2Y receptors in retinal tissue vary according to the species and the cell types. We examined the molecular and pharmacologic profiles of P2Y purinoceptors in retinoblastoma cell, which has not been identified yet. To achieve this goal, we used $Ca^{2+}$ imaging technique and western blot analysis in WERI-Rb-1 cell, a human retinoblastoma cell line. ATP ($10\;{\mu}M$) elicited strong but transient $[Ca^{2+}]_i$ increase in a concentration dependent manner from more than 80% of the WERI-Rb-1 cells (n=46). Orders of potency of P2Y agonists in evoking $[Ca^{2+}]_i$ transients were 2MeS-ATP>ATP>>UTP=${\alpha}{\beta}$-MeATP, which was compatible with the subclass of $P2Y_1$ receptor. The $[Ca^{2+}]_i$ transients evoked by applications of 2MeS-ATP and/or ATP were also profoundly suppressed in the presence of $P2Y_1$ selective blocker (MRS 2179; $30\;{\mu}M$). $P2Y_1$ receptor expression in WERI-Rb-1 cells was also identified by using western blot. Taken together, $P2Y_1$ receptor is mainly expressed in a retinoblastoma cell, which elicits $Ca^{2+}$ release from internal $Ca^{2+}$ storage sites via the phospholipase C-mediated pathway. $P2Y_1$ receptor activation in retinoblastoma cell could be a useful model to investigate the role of purinergic $[Ca^{2+}]_i$ signaling in neural tissue as well as to find a novel therapeutic target to this lethal cancer.

Functional expression of TREK1 channel in human bone marrow and human umbilical cord vein-derived mesenchymal stem cells (사람의 골수와 제대정맥에서 유래된 중간엽 줄기세포에서 TREK1 통로의 기능적 발현)

  • Park, Kyoung Sun;Kim, Yangmi
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.3
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    • pp.1964-1971
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    • 2015
  • Human bone marrow or human umbilical cord vein derived-mesenchymal stem cells (hBM-MSCs or hUC-MSCs) have known as a potentially useful cell type for clinical therapeutic applications. We investigated two-pore domain potassium (K2P) channels in these cells. K2P channels play a major role in setting the resting membrane potential in many cell types. Among them, TREK1 is targets of hydrogen, hypoxia, polyunsaturated fatty acids, antidepressant, and neurotransmitters. We investigated whether hBM-MSCs and hUC-MSCs express functional TREK1 channel using RT-PCR analysis and patch clamp technique. Potassium channel with a single channel conductance of 100 pS was found in hUC-MSCs and BM-MSCs and the channel was activated by membrane stretch (-5 mmHg ~ -15 mmHg), arachidonic acid ($10{\mu}M$) and intracellular acidosis (pH 6.0). These electrophysiological properties were similar to those of TREK1. Our results suggest that TREK1 is functionally present in hBM-MSCs and hUC-MSCs, where they contribute to its resting membrane potential.