• Title/Summary/Keyword: C&RT 방법

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Cloning of An Intron of the Gene Coding for Porcine Acid-Labile Subunit(pALS) of the 150-kDa Insulin-like Growth Factor Complex and the 3' ntranslated Region of pALS Complementary DNA and Confirmation of pALS Gene Expression in Multiple Tissues (돼지 150-kDa Insulin-like Growth Factor Complex의 Acid-labile Subunit(ALS) 유전자의 Intron 및 ALS Complementary DNA의 3' 비해독 부위 Cloning과 생체조직에서의 ALS 유전자 발현 확인)

  • Jin, E.J.;Kim, I.A.;Lee, C.Y.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.555-562
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    • 2004
  • 본 연구는 목저은 다음과 같다: 1) 돼지에서 150-kDa temary insulin-like growth faetor(IGF)complex의 한 구성 요소인 acid-labile subunit(ALS) 유전자 intron의 존재 확인. cloning 및 돼지 ALS(porcine ALS; pALS) complementary DNA(cDNA)의 3' 비해독(untranslated) 부위(3' UT) 증폭. cloning, 2) intron-spanning primer pair를 이용한 reverse transcription-polymerase chain reaction(RT-PCR) 방법에 의한 돼지 조직에서의 ALS 유전자 발현 분포 확인 및 3) 돼지 hepatocyte에서의 ALS 유전자 발현 여부 확인. 돼지 genomic DNA를 template로 하여 PCR 방법으로 예상된는 intron 부위를 증폭하고 plasmid vector에 삽입하여 염기서열을 결정한 결과 타 종의 ALS 유전자에서와 같은 위치에 1,371-base pair(bp)의 pALS intron이 존재함을 확인하였다. 역시 본 연구에서 간에서 추출한 RNA를 주형으로 시작하여 3' rapid amplification of cDNA end(3' RACE) 방법으로 147-bp의 3'UT를 합성하고 그 염기성열을 결정하였다. RT-PCR 결과 간은 물론 조사된 모든 돼지의 내장기관(신장, 폐, 비장)과 자성 생식기관(난소, 난관, 자궁) 및 골격근육에서 ALS 유전자가 발현됨이 밝혀졌다. 또한 돼지 간 조직에 대한 in-situ hybridization 결과 hepatocyte에서 ALS 유전자가 발현됨이 확인되었다. 이상의 결과는 ALS가 혈중 IGF의 저정/조절체로서의 주기능 외에 모세혈관 밖에서도 미지의 기능이 있을 기능성을 시사한다.

Room Acoustic Measurement System Using Impulse Response (임펄스응답을 이용한 실내음향 측정 시스템)

    • The Journal of the Acoustical Society of Korea
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    • v.18 no.5
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    • pp.63-67
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    • 1999
  • Recently, a method of measuring impulse response is widely used for a room acoustic evaluation instead of measuring reverberation time by white noise excitation. Comparing with the traditional reverberation time measurement, this method has many advantages such as good repeatability and the ability to extract various room acoustic parameters at one measurement. In this study, the author developed a measuring system that can extract mono-aural room acoustic parameters from an impulse response measured with MLS (Maximum Length Sequence) signal excitation. These room acoustic parameters include reverberation times(EDT, RT), speech intelligibilities(C50, C80, D, U50, U80, AI) and sound strength(G). This paper introduces the configuration of the developed measuring system, test results and discussions for the measurements at several rooms.

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The Role of c-Jun N-terminal Kinase in the Radiation-Induced Lung Fibrosis (방사선에 의한 폐 섬유화증에서 c-Jun N-terminal Kinase(JNK)의 역할)

  • Uh, Soo-Taek;Hong, Ki-Young;Lee, Young-Mok;Kim, Ki-Up;Kim, Do-Jin;Moon, Seung-Hyuk;Kim, Yong-Hoon;Park, Choon-Sik;Yeom, Uk;Kim, Eun-Suk;Choi, Doo-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.4
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    • pp.450-461
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    • 2001
  • Background : The underlying pathogenesis of radiation-induced lung fibrosis (RTLF) has not been very well defined. However, the role of TGF-$\beta$ in the generation of RTLF has been a major focus because there is an increase in the expression of both the TGF-${\beta}m$-RNA and its protein preceding RTLF lesions. The down stream signal after a TGF-$\beta$ stimulated lung fibrosis includes the activation of many mediators such as Smad and c-Jun N-terminal kinase (JNK) through TAK1. It is we hypothesized that JNK activation may play a pivotal role in RTLF pathogenesis through increased transcription of the fibrogenic cytokines. The present study evaluates JNK activity in alveolar macrophages after irradiation and the relationship between JNK activity and the amount of collagen in the lung tissues. Methods : C57BL/6 mice(20-25 gr, males) received chlorotetracycline(2g/L) in their drinking water 1 week prior to irradiation and continuously there after. The mice were irradiated once with 1400 cGy of $60CO{\gamma}$-ray over the whole chest. The cellular composition of the whole lung bronchoalveoalr lavage fluids(BALF), elastin expression in the lung tissues, the level of hydroxyproline in lung tissues, and an in vitro JNK assay was measured before irradiation and one, four, and eight weeks after irradiation (RT). Results : The volumes of BALF retrieved from instilled 4 mL of saline with 2% heparin were 3.7-3.8 mL for each group. The cell numbers were similar before($4.1{\times}10^4{\pm}0.5{\times}10^4/mL$) and 1 week($3.1{\times}10^4{\pm}0.5{\times}10^4/mL$) after RT. At four and eight weeks after RT, the cell number reached to $14.0{\times}10^4{\pm}1.5{\times}10^4mL$ and $10.0{\times}10^4{\pm}1.3{\times}10^4/mL$, respectively. There we no changes in the lymphocytes and neutrophils population observed in the BALF after RT. The H-E stain of the lung tissues did not show any structural and fibrotic change in the lung tissues at 4 and 8 weeks after RT. In addition, the amount of elastin and collagen were not different on Verhoeff staining of the lung tissues before RT to eight weeks after RT. The hydroxyproine content was measured with the left lung dissected from the left main bronchus. The lung were homogenized and hydrolyzed with 6 N Hel for 12 hours at $110^{\circ}C$ then measured as previously described. The content of hydroxyproline, standardized with a lung protein concentration, reached a peak 4 weeks after RT, and thereafter showed a plateau. AnIn vitro JNK assay using c-$Jun_{1-79}$-GST sepharose beads were performed with the alveolar macrophages obtained from the BAL. JNK activity was not detected prior to RT, However, the JNK activity increased from one week after RT and reached a peak four weeks after RT. Conclusion : JNK may be involved in the pathogenesis because the JNK activity showed similar pattern observed with the hydroxyproine content. However, it is necessary to clarify that the JNK increases the transcription of fibrogenic cyiokines through the transcription factor.

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Transfer and Expression of the Recombinant hFSH Gene in CHO Cells and Transgenic Chickens using Retrovirus Vector System (CHO 세포와 형질전환 닭에 있어서 Retrovirus Vector System에 의한 hFSH 재조합 유전자의 전이와 발현)

  • 권모선;구본철;심호섭;박창식;이성호;김태완
    • Korean Journal of Animal Reproduction
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    • v.27 no.3
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    • pp.197-206
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    • 2003
  • hFSH (human follicle stimulating hormone) is heterodimer consisting of $\alpha$ and $\beta$ subunits. Since assembly of the both subunits in the cell is often the rate-limiting step in production of functional hormone, single-chain hormones have been engineered by genetically linking two different cDNA fragments with a linker sequence. Using retrovirus vector system, the resulting recombinant hFSH gene was transferred in CHO cells and chicken embryos, and the expression of the gene was investigated. In CHO cells, protein synthesis from the single-chain FSH gene was 17 fold higher than that from the heterodimeric counterpart. In the study of transgenic chickens, ten of the eleven chicks hatched from 62 embryos manupulated with recombinant retrovirus stock was determined to carry transgenic genes. RT-PCR analyses confirmed transcription of the single-chain FSH gene, however, no recombinant FSH was detected from the blood samples.

Development and Evaluation of a SYBR Green Real-time PCR Assay for Canine Cytokine Gene Expression (SYBR Green 실시간 역전사 중합효소연쇄반응을 이용한 개 싸이토카인 유전자 발현의 정량)

  • Yu, Do-Hyeon;Ihn, Dong-Chul;Park, Chul;Park, Jin-Ho
    • Journal of Veterinary Clinics
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    • v.27 no.5
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    • pp.508-513
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    • 2010
  • Cytokines are important mediators of the immune response, and quantitating cytokine mRNA is a highly sensitive and attractive method for measuring cytokine production. The objective of the current study was to develop and validate a SYBR green quantitative real-time reverse transcriptase PCR (qRT-PCR) assay for measuring canine cytokine mRNA. The optimal annealing temperatures ($T_a$) of the designed primers were $62^{\circ}C$ for interleukin (IL)-$1{\beta}$, IL-6 and IL-10; $60^{\circ}C$ for glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and tumor necrosis factor (TNF)-${\alpha}$; and $58^{\circ}C$ for high mobility group box 1 (HMGB1). Primer efficiencies of all primers calculated for standard curve samples were between 97.1% and 102.6%. No evidence of secondary structure or primer-dimer formation was seen via melt-curve analysis or gel electrophoresis. The developed qRT-PCR assays are highly specific and sensitive and can be used to quantify gene expression levels of canine cytokines.

Radiation Therapy for Bone Metastases from Hepatocellular Carcinoma: Effect of Radiation Dose Escalation (간세포암에 의한 뼈전이의 방사선치료: 고선량 방사선치료의 효과)

  • Kim, Tae-Gyu;Park, Hee-Chul;Lim, Do-Hoon;Kim, Cheol-Jin;Lee, Hye-Bin;Kwak, Keum-Yeon;Choi, Moon-Seok;Lee, Joon-Hyoek;Koh, Kwang-Cheol;Paik, Seung-Woon;Yoo, Byung-Chul
    • Radiation Oncology Journal
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    • v.29 no.2
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    • pp.63-70
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    • 2011
  • Purpose: To evaluate the extent of pain response and objective response to palliative radiotherapy (RT) for bone metastases from hepatocellular carcinoma according to RT dose. Materials and Methods: From January 2007 to June 2010, palliative RT was conducted for 103 patients (223 sites) with bone metastases from hepatocellular carcinoma. Treatment sites were divided into the high RT dose and low RT dose groups by biologically effective dose (BED) of 39 $Gy_{10}$. Pain responses were evaluated using the numeric rating scale. Pain scores before and after RT were compared and categorized into 'Decreased', 'No change' and 'Increased'. Radiological objective responses were categorized into complete response, partial response, stable disease and progression using modified RECIST (Response Evaluation Criteria In Solid Tumors) criteria; the factors predicting patients' survival were analyzed. Results: The median follow-up period was 6 months (range, 0 to 46 months), and the radiologic responses existed in 67 RT sites (66.3%) and 44 sites (89.8%) in the high and low RT dose group, respectively. A dose-response relationship was found in relation to RT dose (p=0.02). Pain responses were 75% and 65% in the high and low RT dose groups, respectively. However, no statistical difference in pain response was found between the two groups (p=0.24). There were no differences in the toxicity profiles between the high and low RT dose groups. Median survival from the time of bone metastases diagnosis was 11 months (range, 0 to 46 months). The Child-Pugh classification at the time of palliative RT was the only significant predictive factor for patient survival after RT. Median survival time was 14 months under Child-Pugh A and 2 months under Child-Pugh B and C. Conclusion: The rate of radiologic objective response was higher in the high RT dose group. Palliative AT with a high dose would provide an improvement in patient quality of life through enhanced tumor response, especially in patients with proper liver function.

In vivo Radiosensitization Effect of H DAC Inhibitor, SK-7041 on RIF-1 Cell Line (히스톤 탈아세틸효소 억제제 SK-7041의 RIF-1 세포주에 대한 생체내 방사선 감수성 증진 효과)

  • Chie, Eui-Kyu;Shin, Jin-Hee;Kim, In-Ah;Kim, Il-Han
    • Radiation Oncology Journal
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    • v.28 no.4
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    • pp.219-223
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    • 2010
  • Purpose: To test the radiosensitizing effect of the newly synthesized novel histone deacetylase inhibitor, SK-7041 in vivo. Materials and Method: The RIF-l cell line was implanted into the back of a 6-week-old female C3H mouse, intradermally, The mice were grouped into control, drug, radiation (RT), and RT+drug group. SK-7041, 4 mg/kg was administered intraperitoneally for six cycles every 12 hours for mice in the drug and RT+drug group, An identical volume of phosphate buffered saline (PBS) was administered at the same frequency to mice in the control and RT groups. A single 5 Gy fraction was delivered to mice in RT and RT+drug group 6 hours after the fourth delivery. The volume of the implanted tumor was measured every 2~3 days to formulate the growth delay curve. Results: For the control, drug, RT, and RT +drug groups, the average duration for implanted tumor to reach a volume of $1,500mm^3$ was 10 days, 10 days, 9 days, and 12 days, respectively. Moreover, the tumor volume on D14 was $276.7mm^3$, $279.9mm^3$, $292.5mm^3$, and $185.5mm^3$, respectively (p=0.0004). The difference for the change in slope for the control and drug versus the RT and RT+drug groups were borderline significant (p=0.0650). Conclusion: The results of this study indicate that SK-7041 has a radiosensitizing effect for the RIF-1 cell line in vivo at a low concentration and this effect may be synergistic. Implementing this result to clinical trial is warranted.

Development of Protocol for the Effective Detection of Feline Calicivirus as Norovirus Surrogate in Oyster and Lettuce (굴과 상추에서 노로바이러스의 대체모델 feline calicivirus의 효율적 검출법 개발)

  • Lee, Soo-Yeon;Jang, Keum-Il;Woo, Gun-Jo;Kwak, Hyo-Sun;Kim, Kwang-Yup
    • Korean Journal of Food Science and Technology
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    • v.39 no.1
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    • pp.71-76
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    • 2007
  • Foodborne illness caused by Noroviruses (NVs) is increasing rapidly in Korea. This study developed an effective detection protocol for NVs found in contaminated oysters and lettuce through an investigation using the major steps of virus particle separation, concentration and RT-PCR. As a surrogate model for NVs, the cultivable feline calicivirus (FCV) that belongs to the same Caliciviridae family was used. Instead of using a time-consuming ultracentrifugation method, efficient methods based on solvent extraction and PEG precipitation procedure were applied. Direct homogenization of a 25g sample of whole oyster and lettuce in 175mL PBS provided the simplicity that would be needed in the actual field of food product examination. The overnight PEG precipitation step at $4^{\circ}C$ was reduced to 3 h by placing the reaction tube in ice and by adjusting the PEG concentrations. The application of the use of chloroform and 0.2 ${\mu}m$ syringe filtration together showed a better detection efficiency than the use of chloroform alone in removing PCR inhibitors for both oyster and lettuce samples. Also, dilution of the extracted RNA solution before PCR provided increased sensitivity. The improved detection protocol developed in this study could be efficiently applied to detect FCV and most likely NVs from oysters and lettuce.

Concurrent Chemoradiation Therapy in Stage III Non-small Cell Lung Cancer (III 기 비소세포성 폐암에서 Cisplatin-방사선동시병합요법의 효과)

  • Kim In Ah;Choi Ihl Bhong;Kang Ki Mun;Jang Jie Young;Song Jung Sub;Lee Sun Hee;Kuak Mun Sub;Shinn Kyung Sub
    • Radiation Oncology Journal
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    • v.15 no.1
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    • pp.27-36
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    • 1997
  • Purpose : This study was tried to evaluate the Potential benefits of concurrent chemoradiation therapy (low dose daily cisplatin combined with split course radiation therapy) compared with conventional radiation therapy alone in stage III non-small cell lung cancer. The end points of analyses were response rate. overall survival, survival without locoregional failure, survival without distant metastasis, prognostic factors affecting survival and treatment related toxicities. Materials and Methods : Between April 1992 and March 1994, 32 patients who had stage III non-small cell lung cancer were treated with concurrent chemoradiation therapy. Radiation therapy for 2 weeks (300 cGy given 10 times up to 3000 cGy) followed by a 3 weeks rest period and then radiation therapy for 2 more weeks (250 cGy given 10 times up to 2500 cGy) was combined with $6mg/m^2$ of cisplatin. Follow-up period ranged from 13 months to 48 months with median of 24 months. Historical control group consisted of 32 patients who had stage III non-small cell lung cancer were received conventionally fractionated (daily 170-200 cGy) radiation therapy alone. Total radiation dose ranged from 5580 cGy to 7000 cGy with median of 5940 cGy. Follow-up Period ranged from 36 months to 105 months with median of 62 months. Result : Complete reponse rate was higher in chemoradiation therapy (CRT) group than radiation therapy (RT) group (18.8% vs. 6.3%, CRT group showed lower in-field failure rate compared with RT group(25% vs. 47%. The overall survival rate had no significant differences in between CRT group and RT group (17.5% vs. 9.4% at 2 years). The survival without locoregional failure (16.5% vs. 5.3% at 2 years) and survival without distant metastasis (17% vs. 4.6% at 2 years) also had no significant differences. In subgroup analyses for Patients with good performance status (Karnofsky performance scale 80), CRT group showed significantly higher overall survival rate compared with RT group (62.5% vs. 15.6% at 2 years). The prognostic factors affecting survival rate were performance status and pathologic subtype (squamous cell cancer vs. nonsquamous cell cancer) in CRT group. In RT alone group, performance status and stage (IIIa vs IIIb) were identified as a Prognostic factors. RTOG/EORTC grade 2-3 nausea and vomiting(22% vs 6% and bone marrow toxicities (25% vs. 15.6% were significantly higher in CRT group compared with RT alone group. The incidence of RTOG/EORTC grade 3-4 pulmonary toxicity had no significant differences in between CRT group and RT group (16% vs. 6%. The incidence of WHO grade 3-4 pulmonary fibrosis also had no significant differences in both group (38% vs. 25%. In analyses for relationship of field size and Pulmonary toxicity, the Patients who treated with field size beyond 200cm2 had significantly higher rates of pulmonary toxicities. Conclusion : The CRT group showed significantly higher local control rate than RT group. There were no significant differences of survival rate in between two groups. The subgroup of patients who had good performance status showed higher overall survival rate in CRT group than RT group. In spite of higher incidence of acute toxicities with concurrent chemoradiation therapy, the survival gain in subgroup of patients with good performance status were encouraging. CRT group showed higher rate of early death within 1 year, higher 2 year survival rate compared with RT group Therefore, to evaluate the accurate effect on survival of concurrent chemoradiation therapy, systematic follow-up for long term survivors are needed.

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Comparison of Results According to Reaction Conditions of Thyroglobulin Test (Thyroglobulin 검사의 반응조건에 따른 결과 비교 분석)

  • Joung, Seung-Hee;Lee, Young-Ji;Moon, Hyung-Ho;Yoo, So-yoen;Kim, Nyun-Ok
    • The Korean Journal of Nuclear Medicine Technology
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    • v.21 no.1
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    • pp.39-43
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    • 2017
  • Purpose Thyroglobulin (Tg) is a biologic marker of differentiated thyroid carcinoma (DTC), produced by normal thyroid tissue or thyroid cancer tissue. Therefore, the Tg values of DTC patients is the most specific indicator for judging whether recurrence occur or whether the remaining thyroid cancer is present. Thyroid cancer is currently the most common cancer in Korea, of which 90% is differentiated thyroid cancer. The number of patients with thyroid disease of this application also increased, and an accurate and prompt results are required. However, the incubation time of the Tg commonly takes about 24 hours in our hospital, and the result reporting time is delayed, and We could not satisfied with the requirements of clinical departments and patients. In order to fulfill these requirements, experiments were conducted by shortening the incubation time between company B's Kit currently in use and company C's Kit used in other hospitals. Through these experiments, we could perform the correlation with the original method and shortening method, and could find the optimum reaction time to satisfy the needs of the departments and the patients, and we will improve the competitiveness with the EIA examination. Materials and Methods In September 2016, we tested 65 patients company B's kit and company C's kit by three incubation ways. First method $37^{\circ}C$ shaking 2hr/2hr, Second method RT shaking 3hr/2hr, Third method 1hr/1hr shaking at $37^{\circ}C$. Fourth method RT shaking 3hr method which is the original method of Company C's Kit. Fifth method, the incubation time was shortened under room temperature shaking 2hr, Sixth method $37^{\circ}C$ shaking 2hr. And we performed and compared the correlation and coefficient of each methods. Results As a result of performing shortening method on company B currently in use, when comparing the Original method of company B kit, First method $37^{\circ}C$ shaking 2hr/2hr was less than Tg 1.0 ng/mL and the ratio of $R^2=0.5906$, above 1.0 ng/mL In the value, $R^2=0.9597$. Second method RT shaking 3hr/2hr was $R^2=0.7262$ less than value of 1.0 ng/mL, $R^2=0.9566$ above than value of 1.0 ng/mL. Third method $37^{\circ}C$ shaking 1hr/1hr was $R^2=0.7728$ less than value of 1.0 ng/mL, $R^2=0.8904$ above than value of 1.0 ng/mL. Forth, Company C's The original method, RT shaking 3hr was $R^2=0.7542$ less than value of 1.0 ng/mL, and $R^2=0.9711$ above than value of 1.0 ng/mL. Fifth method RT shaking 2hr was $R^2=0.5477$ less than value of 1.0 ng/mL, $R^2=0.9231$ above than value of 1.0 ng/mL. Sixth method $37^{\circ}C$ shaking 2hr showed $R^2=0.2848$ less than value of 1.0 ng/mL, $R^2=0.9028$ above than value of 1.0 ng/mL. Conclusion Samples with both values of 1.0 ng/mL or higher in both of the six methods showed relatively high correlation, but the correlation was relatively low less than value of 1.0 ng/mL. Especially, the $37^{\circ}C$ shaking 2hr method of company C showed a sharp fluctuation from the low concentration value of 1.0 ng/mL or less. Therefore, we are planning to continuously test the time, equipment, incubation temperature and so on for the room temperature shaking 2hr method and $37^{\circ}C$ shaking 1hr/1hr of company C which showed a relatively high correlation. After that, we can search for an appropriate shortening method through additional experiments such as recovery test, dilution test, sensitivity test, and provide more accurate and prompt results to the department of medical treatment, It is competitive with EIA test.

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