• 제목/요약/키워드: reverse transcriptase-polymerase chain reaction (RT-PCR)

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국내 양식 흰다리새우에서 검출된 노랑머리바이러스(YHV-8)의 유전학적 특성 (Detection and genetic characteristic of Yellow-head virus genotype 8 (YHV-8) Cultured Litopanaeus vanamei, in Korea)

  • 김석렬;;;신기욱
    • 한국어병학회지
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    • 제33권1호
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    • pp.77-81
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    • 2020
  • This study was performed for screening yellow head virus(YHV) complex in 252 including 235 white leg shrimps (Litopenaeus vannamei) and 17 oriental shrimp (Fenneropenaeus chinensis) collected from 18 farms located in southwestern province of Korea. The virus complex was detected by nested reverse-transcriptase polymerase chain reaction (RT-PCR) assay. In the assay, amplicons were resulted in RNAs exracted from 38 shrimps (21 white leg shrimps and 17 oriental shrimps) obtained from 7 farms. In phylogenetic analysis using sequences of ORF1b gene, all 38 sequences obtained in this study formed an independent lineage with YHV-8 genotype firstly isolated in China, belonged to an YHV-8 clade.

Protective Effects of Prunus persica Flesh Extract (PPFE) on UV-Induced Oxidative Stress and Matrix Metalloproteinases Expression in Human Skin Cells

  • Park, Hyen-Joo;Park, Kwang-Kyun;Hwang, Jae-Kwan;Chung, Won-Yoon;Kim, Gi-Dae;Lee, Min-Ai;Lee, Sang-Kook
    • Natural Product Sciences
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    • 제18권1호
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    • pp.52-59
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    • 2012
  • In our continuous efforts to procure the active materials from natural products in the protective effects of oxidative stress or UV damage to skin cells we found the Prunus persica flesh extract (PPFE) is considerable to meet the demand to protect the skin damage. PPFE attenuated cell damage induced by hypoxanthine-xanthine oxidase in cultured human keratinocytes, indicating that PPFE has the potential of the scavenging effect of reactive oxygen species (ROS) in human skin cell. Moreover, PPFE significantly suppressed UVA-induced ROS production determined by the oxidation of 2,7-dichlorodihydrofluorescein diacetate (DCFH) using FACS analysis. Additional study revealed that UVA irradiation of HaCaT human keratinocytes increased the gelatinolytic activities of matrix metalloproteinase-2, and -9 (MMP-2, -9) and mRNA expression of MMP-9 analyzing by a real-time reverse transcriptase-polymerase chain reaction (RT-PCR), and these events were significantly suppressed by the treatment with PPFE. These results suggest that PPFE might be applicable as natural ingredients for skin antiaging agents via UV-induced ROS scavenging activity and suppression of MMP expression in the skin cells.

Anti-proliferative and Apoptotic Effects of Dendrosomal Farnesiferol C on Gastric Cancer Cells

  • Aas, Zohreh;Babaei, Esmaeil;Feizi, Mohammad Ali Hosseinpour;Dehghan, Gholamreza
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권13호
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    • pp.5325-5329
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    • 2015
  • Farnesiferol C is a natural compound with various anti-cancer properties that belongs to the class of sesquiterpene coumarins. However, the low bioavailability of farnesiferol C limits its therapeutic potential. Here, we overcame this problem utilizing dendrosome nano-particles and evaluated the anti-cancer effect of dendrosomal farnesiferol C (DFC) on the AGS gastric cancer cell line. The 3-(4,5-dimethyl-thiazol-2yl)-2,5- diphenyl tetrazolium bromide (MTT) assay and reverse transcriptase-polymerase chain reaction (RT-PCR) were respectively used to detect the anti-proliferative properties of DFC and expression ratio of Bax/Bcl-2 as a hallmark of apoptosis. Compared to the void farnesiferol C (FC), our data showed that DFC significantly suppresses the proliferation of AGS cells in a time- and dose-dependent manner (P<0.01). Also, DFC meaningfully increased the expression ratio of Bax/Bcl-2 in AGS cells (P<0.01). The findings demonstrate that our nano-based formulation of farnesiferol C could be considered as a potential therapeutic agent in cancer targeting.

Down-Regulation of CYP1A1 Expression in Breast Cancer

  • Hafeez, S.;Ahmed, A.;Rashid, Asif Z.;Kayani, Mahmood Akhtar
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권5호
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    • pp.1757-1760
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    • 2012
  • Breast cancer is a major cause of death in women worldwide. Mammary tissue expressing xenobiotic metabolizing enzymes metabolically activate or detoxify potential genotoxic breast carcinogens. Deregulation of these xenobiotic metabolizing enzymes is considered to be a major contributory factor to breast cancer. The present study is focused on the expression of the xenobiotic metabolizing gene, CYP1A1, in breast cancer and its possible relationships with different risk factors. Twenty five tumors and twenty five control breast tissue samples were collected from patients undergoing planned surgery or biopsy from different hospitals. Semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and western-blotting were used to investigate the expression of CYP1A1 in breast cancer control and disease samples. mRNA expression of CYP1A1 was down-regulated in 40% of breast tumor samples. Down-regulation was also observed at the protein level. Significnat relations were noted with marital status and tumour grade but not histopathological type. In conclusion, CYP1A1 protein expression was markedly reduced in tumor breast tissues samples as compared to paired control tissue samples.

국내분리 소 로타바이러스 NSP4 유전자의 크로닝 및 염기서열 분석 (Cloning and Sequencing of Nonstructural Protein (NSP4) Gene of Bovine Rotavirus Isolated in Korea)

  • 백명순;김원용;강신영
    • 대한바이러스학회지
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    • 제30권1호
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    • pp.1-10
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    • 2000
  • The nonstructural protein NSP4, encoded by gene 10 of rotavirus, has been shown to playa role in viral assembly and known to be an enterotoxin, causing diarrhea in mouse pups. NSP4 gene was cloned from CBNU-2 (virulent bovine rotavirus/diarrheic fecal sample) and CBNU-1 (cell-culture adapted bovine rotavirus/isolated from CBNU-2 and 75 times passaged on MA104 cells), respectively, by reverse transcriptase-polymerase chain reaction (RT-PCR) and sequenced and compared. The sequence data indicated that the NSP4 genes of bovine rotavirus (BRV) were 751 bases in length and encoded one open reading frame of 175 amino acids beginning at base 42 and terminating at base 569. Differences in nucleotide sequence between CBNU-2 and CBNU-1 were observed at 6 positions (base 274, 296, 391, 394, 396 and 579). NSP4 gene of BRV exhibited a high degree of nucleotide (90% and 94%) and amino acid sequence (91% and 97%) homology with those of SA11 and UK but a low degree of nucleotide (77% and 79%) and amino acids sequence (81% and 85%) homology with those of Wa and OSU.

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Inhibition of Cervical Cancer Cell Growth by Gene Silencing of HPV16 E6 Induced by Short-interfering RNA

  • Park, Sang-Muk;Lee, Sun-Kyung;Kim, Yoon-Sik
    • 대한임상검사과학회지
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    • 제43권3호
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    • pp.89-97
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    • 2011
  • The Human Papilloma Virus (HPV) infection has been strongly associated with pathogenesis of uterine cervix carcinoma. HPV type 16, a causative agent of uterine cervix carcinoma, encodes the E6 and E7 oncogenes, expression of which is pivotal for malignant transformation and maintenance of malignant phenotypes. To develop a gene therapy for HPV-related carcinoma, We investigated the effect of E6 short-interfering RNA (E6 siRNA) on the expression of this oncogene and on the growth of HPV 16-related uterine cervix carcinoma cells. SiHa cells, a uterine cervix carcinoma cell line, which contain a single copy of HPV 16 integrated in the chromosome and express the E6 and E7 oncogenes. Before 24 hr of transfection, cells were seeded and transfected with control plasmid or E6 siRNA-expressing plasmid. The mRNA was analysed by reverse transcriptase polymerase chain reaction (RT-PCR). The cell growth rate was investigated by MTT method. The E6 mRNA level in SiHa cells was decreased in HPV 16 E6 siRNA-expression vector transfected cells and a decrease in the growth of these cells was also observed. From these results. it is evident that E6 siRNA played a role in suppression of growth of SiHa cells and has a fair chance as a candidate for gene specific therapy for HPV related uterine cervix carcinoma.

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미강에탄올추출물의 RAW264.7 세포에서 항염증효과 (Anti-Inflammatory Effects of Rice Bran Ethanol Extract in Murine Macrophage RAW 264.7 Cells)

  • 박정숙;김미혜
    • 약학회지
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    • 제55권6호
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    • pp.456-461
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    • 2011
  • The aim of the present study is to investigate the anti-inflammatory effect of a Rice Bran Ethanol Extract (RBE). Inflammation, such as a bacterial infection in vivo metabolites, such as external stimuli or internal stimuli to the defense mechanisms of the biological tissue a variety of intracellular regulatory factors deulin inflammatory TNF-${\alpha}$, IL-$1{\beta}$, IL-6, IL-8, such as proinflammatory cytokines, prostagrandin, lysosomal enzyme, free radicals are involved in a variety of mediators. The present study was designed to determine the effect of the RBE on pro-inflammatory factors such as NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 in lipopolysaccharide (LPS) - stimulated RAW264.7 macrophages cells. The cell toxicity was determined by MTS assay. To evaluate of anti-inflammatory effect of RBE, amount of NO was measured using the NO detection kit and the iNOS expression was measured by reverse transcriptase polymerase chain reaction (RT-PCR). And proinflammatory cytokines were measured by ELISA kit. As a result, the RBE reduced NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 production without cytotoxicity. Our results suggest that the RBE may have an anti-inflammatory property through suppressing inflammatory mediator productions and appears to be useful as an anti-inflammatory material.

Isolation and Characterization of Terpene Synthase Gene from Panax ginseng

  • Kim, Yu-Jin;Ham, Ah-Rom;Shim, Ju-Sun;Lee, Jung-Hye;Jung, Dae-Young;In, Jun-Gyo;Lee, Bum-Soo;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • 제32권2호
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    • pp.114-119
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    • 2008
  • Terpene synthase plays a key role in biosynthesis of triterpene saponins (ginsenosides) and is intermediate in the biosynthesis of a number of secondary metabolites. A terpene synthase (PgTPS) cDNA was isolated and characterized from the root of Panax ginseng c.A. Meyer. The deduced amino acid sequence of PgTPS showed a similarity with A. deliciosa (AAX16121) 61%, V. vinifera (AAS66357) 61%, L. hirsutum (AAG41891) 55%, M. truncatula (AAV36464) 52%. And the segment of a terpene synthase gene was amplified by reverse transcriptase-polymerase chain reaction (RTPCR). We studied expression of terpene synthase under stressful conditions like chilling, salt, UV, and heavy metal stress treatment. Expression of PgTPS was increased gradually after exposure to stresses such as chilling, salt, and UV illumination. But its transcription seems to be reduced by cadmium and copper treatment.

오레가노 추출물의 모발성장 촉진효과 (Effect of Origanum vulgare Extracts on Hair Regeneration)

  • 박장순
    • 생약학회지
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    • 제44권3호
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    • pp.275-280
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    • 2013
  • This study was carried out to investigate the effect of Origanum vulgare extracts on cell proliferation of human hair dermal papilla cell (HHDPC) using sulforhodamine B (SRB) assay, antioxidant activity by 1,1-diphenyl-2-picryl hydrazyl (DPPH) method, expression of insulin-like growth factor-1 (IGF-1) by analyzing reverse transcriptase polymerase chain reaction (RT-PCR) and hair growth in a shaving animal model of C57BL/6 mice topically applying with an amount of 0.1 mL once a day for 3 weeks. The mice were divided into 4 groups including normal group (saline, N), negative control group (dimethyl sulfoxide, NC), positive control group (5 mg/mL minoxidil, PC), and experimental group (Origanum vulgare extracts, OV). Treatment of OV didn't show cytotoxicity in HHDPC up to 10 ${\mu}g/mL$ and exhibited antioxidant activity with $IC_{50}$ of 31.0 ${\mu}g/mL$. IGF-1 expression in the skin was significantly (p<0.05) increased in the PC and OV compared to the N or NC. PC and OV also showed a prominently promoted hair regrowth compared to the N or NC in hair growth observation. The hair regrowth of OV was significantly higher than that of PC (p<0.05). Therefore, these results indicate that O. vulgare extracts effectively stimulated hair growth in an animal model.

Sandalwood Essential Oil의 iNOS 발현과 염증성사이토카인의 생성에 미치는 영향 (Effects of Sandalwood Essential Oil on the iNOS Expression and Proinflammatory Cytokine Production)

  • 박정숙;정숙희
    • 약학회지
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    • 제57권1호
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    • pp.70-75
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    • 2013
  • The present study was designed to determine the effect of the Sandalwood Essential Oil (Santalum album) on pro-inflammatory factors such as NO, iNOS expression and IL-$1{\beta}$, IL-6, TNF-${\alpha}$ in lipopolysaccharide (LPS) - stimulated RAW264.7 macrophages cells. The cell toxicity was determined by MTS assay. To evaluate of anti-inflammatory effect of Sandalwood Essential Oil, amount of NO was measured using the NO detection kit and the iNOS expression was measured by western blot analysis and reverse transcriptase polymerase chain reaction (RT-PCR). And proinflammatory cytokines were measured by ELISA kit. As a result, Sandalwood Essential Oil reduced NO, iNOS expression and IL-$1{\beta}$, IL-6, TNF-${\alpha}$ production without cytotoxicity. Our results suggest that the Sandalwood Essential Oil may have an anti-inflammatory property through suppressing inflammatory mediator productions and appears to be useful as an anti-inflammatory oil.