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Tumorigenic Effects of Endocrine-Disrupting Chemicals are Alleviated by Licorice (Glycyrrhiza glabra) Root Extract through Suppression of AhR Expression in Mammalian Cells

  • Chu, Xiao Ting;Cruz, Joseph Dela;Hwang, Seong Gu;Hong, Heeok
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.13
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    • pp.5117-5121
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    • 2014
  • Endocrine-disrupting chemicals (EDCs) have been reported to interfere with estrogen signaling. Exposure to these chemicals decreases the immune response and causes a wide range of diseases in animals and humans. Recently, many studies showed that licorice (Glycyrrhiza glabra) root extract (LRE) commonly called "gamcho" in Korea exhibits antioxidative, chemoprotective, and detoxifying properties. This study aimed to investigate the mechanism of action of LRE and to determine if and how LRE can alleviate the toxicity of EDCs. LRE was prepared by vacuum evaporation and freeze-drying after homogenization of licorice root powder that was soaked in 80% ethanol for 72 h. We used 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) as an EDC, which is known to induce tumors or cancers; MCF-7 breast cancer cells were used as a tumorigenic model. These were treated with TCDD and various concentrations of LRE (0, 50, 100, 200, $400{\mu}g/mL$) for 24, 48, and 72 h. As a result, TCDD stimulated MCF-7 cell proliferation, but LRE significantly inhibited TCDD-induced MCF-7 cell proliferation in a dose- and time-dependent manner. Expression of TCDD toxicity-related genes, i.e., aryl hydrocarbon receptor (AhR), AhR nuclear translocator, and cytochrome P450 1A1, were subsequently down-regulated by LRE in a dose-dependent manner. Analysis of cell cycle distribution after treatment of MCF-7 cells with TCDD and various concentrations of LRE showed that LRE inhibited the proliferation of MCF-7 cells via G2/M phase arrest. Reverse transcription-polymerase chain reaction and Western blot analyses also revealed that LRE dose-dependently increased the expression of the tumor suppressor genes p53 and p27 and down-regulated the expression of cell cycle-related genes. These data suggest that LRE can mitigate the tumorigenic effects of TCDD in breast cancer cells by suppression of AhR expression and cell cycle arrest. Thus, LRE can be used as a potential toxicity-alleviating agent against EDC-mediated disease.

MicroRNA-101 Inhibits Cell Proliferation, Invasion, and Promotes Apoptosis by Regulating Cyclooxygenase-2 in Hela Cervical Carcinoma Cells

  • Huang, Fei;Lin, Chen;Shi, Yong-Hua;Kuerban, Gulinar
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5915-5920
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    • 2013
  • Aim: Although aberrant miRNA expression has been documented, altered miR-101 expression in cervical cancer and its carcinogenic effects and mechanisms remain unexplored. The aim of our study was to investigate the role of miR-101 alteration in cervical carcinogenesis. Methods: Expression of miR-101 was examined by quantitative real-time reverse transcriptase PCR (qRT-PCR) in Hela cells. After modulating miR-101 expression using miR-101 mimics, cell growth, apoptosis and proliferation, and migration were tested separately by MTT or flow cytometry and cell wound healing assay and protein expression was detected by qRT-PCR. The expression of COX-2 in Hela cell was also examined by immunohistochemical staining and the correlation with miR-101 expression was analysed. Results: The miR-101 demonstrated significantly low expression in Hela cell. When we transfected miR-101 mimics into Hela cells, the modulation of miR-101 expression remarkably influenced cell proliferation, cycling and apoptosis: 1) The expression of microRNA-101 tended to increase after transfection; 2) Overexpression of miR-101 was able to promote cell apoptosis, the apoptosis rate being markedly higher (97.6%) than that seen pre-transfection (12.2%) (P<0.05); 3) The miR-101 negatively regulates cell migration and invasion, scratch results being lower ($42.7um{\pm}2um$) than that observed pre-transfection ($181.4um{\pm}2um$); 4) miRNA-101 inhibits the proliferation of Hela cells as well as the level of COX-2 protein, which was negatively correlated with miR-101 expression. Conclusions: Overexpression of miR-101 has obvious inhibitory effects on cell proliferation, migration and invasion. Thus reduced miR-101 expression could participate in the development of cervical cancer at least partly through loss of inhibition of target gene COX-2, which probably occurs in a relative late phase of carcinogenesis. Our data suggest an important role of miR-101 in the molecular etiology of cancer and indicate potential application of miR-101 in cancer therapy.

CCNG2 Suppressor Biological Effects on Thyroid Cancer Cell through Promotion of CDK2 Degradation

  • Li, Wei-Juan;Liu, Ge-Ling;Yu, Fang;Xiang, Xiu-Xiu;Lu, Yi-Fang;Xiao, Hong-Zhen;Shi, Yan-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.6165-6171
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    • 2013
  • This study aimed to analyze the expression and clinical significance of cyclin G2 (CCNG2) in thyroid carcinoma and the biological effects of CCNG2 overexpression in a cell line. Immunohistochemistry and Western blotting were used to analyze CCNG2 protein expression in 63 cases of thyroid cancer and normal tissues to allow the relationship with clinical factors to be assessed. CCNG2 lentiviral and empty vectors were transfected into the thyroid cancer K1 cell line. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were applied to detect the mRNA and protein levels of CCNG2. MTT assay and cell cycle were also conducted to assess the influence of up-regulated expression of CCNG2 on K1 cell biology. The level of CCNG2 protein expression was found to be significantly lower in thyroid cancer tissue than normal tissues (P<0.05). Western blot: The relative amount of CCNG2 protein in thyroid cancer tissue was respectively found to be significantly lower than in normal tissues (P<0.05), correlating with lymph node metastasis, clinic stage and histological grade (P<0.05), but not gender, age or tumor size (P>0.05). Loss of CCNG2 expression correlated significantly with poor overall survival time on Kaplan-Meier analysis (P<0.05). The results for biological functions showed that K1 cell transfected CCNG2 had a lower survival fraction, a greater percentage in the G0/G1 phases, and lower cyclin-dependent kinase 2 (CDK2) protein expression compared with K1 cells non-transfected with CCNG2 (P<0.05). CCNG2 expression decreased in thyroid cancer and correlated significantly lymph node metastasis, clinic stage, histological grade and poor overall survival, suggesting that CCNG2 may play important roles as a negative regulator in thyroid cancer K1 cells by promoting degradation of CDK2.

Development of Leg Stiffness Controllable Artificial Tendon Actuator (LeSATA®) Part I - Gait Analysis of the Metatarsophalangeal Joint Tilt Angles Soonhyuck - (하지강성 가변 인공건 액추에이터(LeSATA®)의 개발 Part I - Metatarsophalangeal Joint Tilt Angle의 보행분석 -)

  • Han, Gi-Bong;Eo, Eun-Kyung;Oh, Seung-Hyun;Lee, Soon-Hyuck;Kim, Cheol-Woong
    • Transactions of the KSME C: Technology and Education
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    • v.1 no.2
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    • pp.153-165
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    • 2013
  • The established gait analysis studies have regarded leg as one single spring. If we can design a knee-ankle actuating mechanism as a primary actuator for supporting knee extension, it might be possible to revolutionary store or release elastic strain energy, which is consumed during the gait cycle, and as a result leg stiffness is expected to increase. An ankle joint actuating mechanism that stores and releases the energy in ankle joint is expected to support and solve excessive artificial leg stiffness caused by the knee actuator (primary actuator) to a reasonable extent. If unnecessary kinematic energy is released with the artificial speed reduction control designed to prevent increase in gait speed caused by increase in time passed, it naturally brings question to the effectiveness of the actuator. As opposed to the already established studies, the authors are currently developing knee-ankle two actuator system under the concept of increasing lower limb stiffness by controlling the speed of gait in relative angular velocity of the two segments. Therefore, the author is convinced that compensatory mechanism caused by knee actuating must exist only in ankle joint. Ankle joint compensatory mechanism can be solved by reverse-examining the change in metatarso-phalangeal joint (MTPJ) tilt angle (${\theta}_1=0^{\circ}$, ${\theta}_2=17^{\circ}$, ${\theta}_3=30^{\circ}$) and the effect of change in gait speed on knee activity.

Optimization of Reference Genes for Normalization of the Quantitative Polymerase Chain Reaction in Tissue Samples of Gastric Cancer

  • Zhao, Lian-Mei;Zheng, Zhao-Xu;Zhao, Xiwa;Shi, Juan;Bi, Jian-Jun;Pei, Wei;Feng, Qiang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.14
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    • pp.5815-5818
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    • 2014
  • For an exact comparison of mRNA transcription in different samples or tissues with real time quantitative reverse transcription-polymerase chain reaction (qRT-PCR), it is crucial to select a suitable internal reference gene. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and beta-actin (ACTB) have been frequently considered as house-keeping genes to normalize for changes in specific gene expression. However, it has been reported that these genes are unsuitable references in some cases, because their transcription is significantly variable under particular experimental conditions and among tissues. The present study was aimed to investigate which reference genes are most suitable for the study of gastric cancer tissues using qRT-PCR. 50 pairs of gastric cancer and corresponding peritumoral tissues were obtained from patients with gastric cancer. Absolute qRT-PCR was employed to detect the expression of GAPDH, ACTB, RPII and 18sRNA in the gastric cancer samples. Comparing gastric cancer with corresponding peritumoral tissues, GAPDH, ACTB and RPII were obviously upregulated 6.49, 5.0 and 3.68 fold, respectively. Yet 18sRNA had no obvious expression change in gastric cancer tissues and the corresponding peritumoral tissues. The expression of GAPDH, ${\beta}$-actin, RPII and 18sRNA showed no obvious changes in normal gastric epithelial cells compared with gastric cancer cell lines. The carcinoembryonic antigen (CEA), a widely used clinical tumor marker, was used as a validation gene. Only when 18sRNA was used as the normalizing gene was CEA obviously elevated in gastric cancer tissues compared with peritumoral tissues. Our data show that 18sRNA is stably expressed in gastric cancer samples and corresponding peritumoral tissues. These observations confirm that there is no universal reference gene and underline the importance of specific optimization of potential reference genes for any experimental condition.

$CF_4$/Ar를 이용한 유기고분자 기판의 펄스 직류전원 건식 식각

  • Kim, Jin-U;Choe, Gyeong-Hun;Park, Dong-Gyun;Jo, Gwan-Sik;Lee, Je-Won
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.08a
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    • pp.91-91
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    • 2010
  • 본 논문은 펄스 직류전원 (Pulse DC) 플라즈마 소스와 반응성 가스인 $CF_4$와 불활성 가스인 Ar를 혼합하여 산업에서 널리 사용되는 유기고분자인 Polymethylmethacrylate (PMMA), Polyethylene terephthalate (PET), 그리고 Polycarbonate (PC) 샘플을 건식 식각한 결과에 대한 것이다. 각각의 샘플은 감광제 도포 후에 자외선을 조사하는 포토레지스트 방법으로 마스크를 만들었다. 펄스 직류전원 플라즈마 시스템을 사용하면 다양한 변수를 줄 수 있다는 장점이 있다. 공정 변수는 Pulse DC Voltage는 300 - 500 V, Pulse DC reverse time $0.5{\sim}2.0\;{\mu}s$, Pulse DC Frequency 100~250 kHz 이었다. 변수 각각의 값이 높아질수록 고분자의 식각률이 높아졌다. 특히, PMMA의 식각률이 가장 높았으며 PET, PC 순이었다. 샘플 중 PC의 식각률이 가장 낮은 이유는 고분자 결합 중에 이중결합의 벤젠 고리 모양을 포함하고 있어 분자 결합력이 비교적 높기 때문으로 사료된다. 기계적 펌프만을 사용한 공정 전 압력은 30 mTorr이었다. 쓰로틀 밸브를 완전 개방한 상태에서 식각 공정 중 진공 압력은 $CF_4$ 가스유량이 늘어날수록 증가하였다. 식각률 역시 $CF_4$ 가스유량(총 가스 유량은 10 sccm)이 많을수록 증가함을 보여주었다 (PMMA: 10 sccm $CF_4$에서 330 nm/min, 3.5 sccm $CF_4$/6.5 sccm Ar에서 260 nm/min., PET: 10 sccm $CF_4$에서 260 nm/min, 3.5 sccm $CF_4$/6.5 sccm Ar에서 210 nm., PC: 10 sccm $CF_4$에서 230 nm, 3.5 sccm $CF_4c$/6.5 sccm Ar에서 160 nm). 이는 펄스 직류전원 플라즈마 식각에서 $CF_4$와 Ar의 가스 혼합비를 조절함으로서 고분자 소재의 식각률을 적절히 변화시킬 수 있다는 것을 의미한다. 표면 거칠기는 실험 후 표면단차 측정기와 전자 현미경 등을 이용하여 식각한 샘플의 표면을 측정하여 알 수 있었다. 실험전 기준 샘플 표면 거칠기는 PMMA는 1.53nm, PET는 3.1nm, PC는 1.54nm 이었다. 식각된 샘플들의 표면 거칠기는 PMMA는 3.59~10.59 nm, PET은 5.13~11.32 nm, PC는 1.52~3.14 nm 범위였다. 광학 발광 분석기 (Optical emission spectroscopy)를 이용하여 식각 공정 중 플라즈마 발광특성을 분석한 결과, 탄소 원자 픽 (424.662 nm)과 아르곤 원자 픽 (751.465 nm, 763.510 nm)의 픽의 존재를 확인하였다. 이 때 탄소 픽은 $CF_4$ 가스에서 발생하였을 것으로 추측한다. 본 발표를 통해 펄스 직류전원 $CF_4$/Ar의 고분자 식각 결과에 대해 보고할 것이다.

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Correlation between Changes in Microbial/Physicochemical Properties and Persistence of Human Norovirus during Cabbage Kimchi Fermentation

  • Lee, Hee-Min;Lee, Ji-Hyun;Kim, Sung Hyun;Yoon, So-Ra;Lee, Jae Yong;Ha, Ji-Hyoung
    • Journal of Microbiology and Biotechnology
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    • v.27 no.11
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    • pp.2019-2027
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    • 2017
  • Recently, cabbage kimchi has occasionally been associated with the foodborne diseases of enteric viruses such as human norovirus (HuNoV). This study aimed to evaluate the correlation between microbial/physicochemical properties and persistence of HuNoV in experimentally contaminated cabbage kimchi fermented and stored at $4^{\circ}C$ or $10^{\circ}C$ for 28 days. Changes in organic acid content, lactic acid bacteria (LAB), acidity, pH, and salinity were analyzed. The recovery of structurally intact HuNoV was examined for up to 28 days post-inoculation, using a NoV GII.4 monoclonal antibody-conjugated immuno-magnetic separation method combined with quantitative real-time reverse transcription polymerase chain reaction. On day 0, LAB loads were $4.70log_{10}$ colony forming units/g and HuNoV GII.4 titers were $2.57log_{10}\;genomic\;copies/{\mu}l$, at both temperatures. After 28 days, intact HuNoV titers decreased to 1.58 ($4^{\circ}C$) and $1.04(10^{\circ}C)log_{10}\;genomic\;copies/{\mu}l$, whereas the LAB density increased. This correlated with a gradual increase in lactic acid and acetic acid at both temperatures. Our findings support a statistical correlation between changes in physicochemical properties and the recovery of structurally intact HuNoV GII.4. Moreover, we determined that the production of organic acid and low pH could affect HuNoV GII.4 titers in cabbage kimchi during fermentation. However, HuNoV GII.4 was not completely eliminated by microbial/physicochemical factors during fermentation, although HuNoV GII.4 was reduced. Based on this, we speculate that the persistence of HuNoV GII.4 may be affected by the continually changing conditions during kimchi fermentation.

Fabrication and Characteristics of a Varactor Diode for UHF TV Tuner Operated within Low Tuning Voltage (저전압 UHF TV 튜너용 바렉터 다이오드의 제작 및 특성)

  • Kim, Hyun-Sik;Moon, Young-Soon;Son, Won-Ho;Choi, Sie-Young
    • Journal of Sensor Science and Technology
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    • v.23 no.3
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    • pp.185-191
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    • 2014
  • The width of depletion region in a varactor diode can be modulated by varying a reverse bias voltage. Thus, the preferred characteristics of depletion capacitance can obtained by the change in the width of depletion region so that it can select only the desirable frequencies. In this paper, the TV tuner varactor diode fabricated by hyper-abrupt profile control technique is presented. This diode can be operated within 3.3 V of driving voltage with capability of UHF band tuning. To form the hyperabrupt profile, firstly, p+ high concentration shallow junction with $0.2{\mu}m$ of junction depth and $1E+20ions/cm^3$ of surface concentration was formed using $BF_2$ implantation source. Simulation results optimized important factors such as epitaxial thickness and dose quality, diffusion time of n+ layer. To form steep hyper-abrupt profile, Formed n+ profile implanted the $PH_3$ source at Si(100) n-type epitaxial layer that has resistivity of $1.4{\Omega}cm$ and thickness of $2.4{\mu}m$ using p+ high concentration Shallow junction. Aluminum containing to 1% of Si was used as a electrode metal. Area of electrode was $30,200{\mu}m^2$. The C-V and Q-V electric characteristics were investigated by using impedance Analyzer (HP4291B). By controlling of concentration profile by n+ dosage at p+ high concentration shallow junction, the device with maximum $L_F$ at -1.5 V and 21.5~3.47 pF at 0.3~3.3 V was fabricated. We got the appropriate device in driving voltage 3.3 V having hyper-abrupt junction that profile order (m factor) is about -3/2. The deviation of capacitance by hyper-abrupt junction with C0.3 V of initial capacitance is due to the deviation of thermal process, ion implantation and diffusion. The deviation of initial capacitance at 0.3 V can be reduced by control of thermal process tolerance using RTP on wafer.

A Study on Diagnostic Criteria of Noise-Induced Hearing Loss among Workers in an Iron Foundry (철강공장 근로자를 대상으로 살펴본 소음성 난청 진단기준에 관한 조사)

  • Kim, Ji-Yong;Lim, Hyun-Sul;Cheong, Hae-Kwan;Moon, Ok-Ryun
    • Journal of Preventive Medicine and Public Health
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    • v.26 no.3 s.43
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    • pp.371-386
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    • 1993
  • This study was carried out to evaluate diagnostic criteria of noise-induced hearing loss (NIHL) among-workers in an iron foundry. Of 1,093 workers under the observation of noise-specific health examination, 184 workers were selected by way of first and second screening audiometric examination. A questionnaire survey, otological examinations, Rinne test and audiometric test were performed and the results were as follows ; The degree of hearing impairment in the left ear was more severe than in the right ear (p<0.05). The difference between hearing threshold of the first and the second hearing test at 1,000 Hz was about 5 dB with a narrow range of deviations while the difference at 4,000 Hz was about -7 dB with a wide range. Of the total study workers, 84.8% were tested within 15 hours away from noise exposure, and the rest after 16 hours. This study has identified that mean hearing loss at 4,000 Hz showed a significant statistical difference among the two study groups while mean hearing loss by 4-divided classification did not. The same phenomena were observed between the group with and without tinnitus and between the group with and without difficulty in hearing (p<0.05). Among 184 workers, 10 workers (5.4%) diagnosed as NIHL by old diagnostic criteria in contrast to 150 workers diagnosed as NIHL by the new diagnostic criteria. There was a significant difference between the two groups in the average hearing loss at 4,000 Hz and 4-divided classification (p<0.01), but there were no significant differences in age, the duration of employment, blood pressure and the duration wearing the personal hearing protector (p>0.05). If we apply Early Loss Index (ELI) method, some workers in younger age group diagnosed as NIHL by the new diagnostic criteria were fallen into within the normal range. In the mean time older age group show reverse results in contrast to the above finding. It is too early to confirm the value of the usage of the new diagnostic criteria in hearing examination. Further study is called for to verify the value of this criteria.

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Effect of 18 Irradiation on Neurotransmitters in the Brains of Goldfish Carassius auratus (18 방사성동위원소 피폭에 의한 금붕어(Carassius auratus)뇌의 신경전달물질 변화)

  • Park, Nam-Gyu;Go, Hye-Jin;Kim, Gun-Do;Lee, Jong-Kyu;Kil, Sang-Hyeong;Lee, Byung-Woo
    • Journal of Life Science
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    • v.22 no.8
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    • pp.1046-1051
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    • 2012
  • In order to investigate the changes in bioactive materials induced in goldfish brains by $^{18}F$ irradiation, the variations in the neurotransmitter levels in the whole brain were studied. The distance between the goldfish and 580 mCi of $^{18}F$ was about 4 cm, and the exposure lasted for 4 hrs. The absorption level calculated based on the distance, exposure time, and half-life of $^{18}F$ was approximately 2 Gy. After sacrifice by $^{18}F$ irradiation or untreated conditions, ten brains were dissected or immediately frozen, respectively. The tissues were extracted in acetic acid. After lyophilization, the samples were dissolved in distilled water and were further purified on a reverse-phase HPLC column. There were no differences in the intensities of the bioactive materials between $^{18}F$-exposed goldfish and control goldfish, while the only peak corresponded to 13 min, which indicated a significant increase in the irradiated brains. Our analysis has found that this compound is tryptophan. This result suggests that $^{18}F$ leads to changes in a classical neurotransmitter, tryptophan, in both the brains of control goldfish and goldfish contaminated by irradiation.