• 제목/요약/키워드: Fluorescence In situ Hybridization

검색결과 206건 처리시간 0.025초

Significance of Human Telomerase RNA Gene Amplification Detection for Cervical Cancer Screening

  • Chen, Shao-Min;Lin, Wei;Liu, Xin;Zhang, You-Zhong
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권5호
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    • pp.2063-2068
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    • 2012
  • Aim: Liquid-based cytology is the most often used method for cervical cancer screening, but it is relatively insensitive and frequently gives equivocal results. Used as a complementary procedure, the high-risk human papillomavirus (HPV) DNA test is highly sensitive but not very specific. The human telomerase RNA gene (TERC) is the most often amplified oncogene that is observed in cervical precancerous lesions. We assessed genomic amplification of TERC in liquid-based cytological specimens to explore the optimal strategy of using this for cervical cancer screening. Methods: Six hundred and seventy-one residual cytological specimens were obtained from outpatients aged 25 to 64 years. The specimens were evaluated by the Digene Hybrid Capture 2 (HC2) HPV DNA test and fluorescence in situ hybridization (FISH) with a chromosome probe to TERC (3q26). Colposcopic examination and histological evaluation were performed where indicated. Results: The TERC positive rate was higher in the CIN2+ (CIN2, CIN3 and SCC) group than in the normal and CIN 1 groups (90.0% vs. 10.4%, p < 0.01). In comparison with the HC2 HPV DNA test, the TERC amplification test had lower sensitivity but higher specificity (90.0% vs. 100.0%, 89.6% vs. 44.0%, respectively). TERC amplification test used in conjunction with the HC2 HPV DNA test showed a combination of 90.0% sensitivity and 92.2% specificity. Conclusion: The TERC amplification test can be used to diagnose cervical precancerous lesions. TERC and HPV DNA co-testing shows an optimal combination of sensitivity and specificity for cervical cancer screening.

호밀(Secale cereale L.)의 핵형분석과 rDNA의 Physical Mapping (Karyotype Analysis and rDNA Physical Mapping in Rye (Secale cereale L.))

  • 이준수;서봉보;김민
    • 한국육종학회지
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    • 제42권2호
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    • pp.163-168
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    • 2010
  • 본 실험은 곡류 작물중에서 육종의 소재로써 많은 장점을 지닌 호밀을 Gimsa C-분염법과 FISH기법을 이용하여 구성이질 염색질과 5S와 18S-26S rRNA 유전자의 염색체상의 위치를 확인하고자 본 실험을 수행하였다. 그 결과를 요약하면 아래와 같다. 표지되었으며 2차협착으로 부수체가 존재하는 1번 염색체의 부수체의 말단과 5번 염색체의 중간에 표지되었고, 18S-26S rDNAs 유전자는 1개의 염색체에 표지되었으며 이 염색체는 2차협착으로 부수체가 존재하는 1번 염색체의 인 형성체 부위에 표지되었다. 1번 염색체에는 5S 와 18S-26S rDNAs 유전자가 표지되었고 5번 염섹체에는 5S rDNA 유전자만이 표지되었다.

Altered expression of MALAT1 lncRNA in chronic lymphocytic leukemia patients, correlation with cytogenetic findings

  • Ahmadi, Abdolrahim;Kaviani, Saeid;Yaghmaie, Marjan;Pashaiefar, Hossein;Ahmadvand, Mohammad;Jalili, Mahdi;Alimoghaddam, Kamran;Eslamijouybari, Mohammad;Ghavamzadeh, Ardeshir
    • BLOOD RESEARCH
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    • 제53권4호
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    • pp.320-324
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    • 2018
  • Background Recent studies have devoted much attention to non-protein-coding transcripts in relation to a wide range of malignancies. MALAT1, a long non-coding RNA, has been reported to be associated with cancer progression and prognosis. Thus, we here determined MALAT1 gene expression in chronic lymphocytic leukemia (CLL), a genetically heterogeneous disease, and explored its possible relationships with cytogenetic abnormalities. Methods MALAT1 expression level was evaluated using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) on blood mononuclear cells from 30 non-treated CLL patients and 30 matched healthy controls. Cytogenetic abnormalities were determined in patients by fluorescence in situ hybridization (FISH). Results MALAT1 expression level was up-regulated in the CLL group compared to healthy controls (P=0.008). Del13q14, followed by Del11q22, were the most prevalent cytogenetic abnormalities. We found no association between the FISH results and MALAT1 expression in patients. Conclusion Altered expression of MALAT1 is associated with CLL development. Further investigations are required to assess the relationship between this long non-coding RNA and CLL patient survival and prognosis.

CircCOL1A2 Sponges MiR-1286 to Promote Cell Invasion and Migration of Gastric Cancer by Elevating Expression of USP10 to Downregulate RFC2 Ubiquitination Level

  • Li, Hang;Chai, Lixin;Ding, Zujun;He, Huabo
    • Journal of Microbiology and Biotechnology
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    • 제32권7호
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    • pp.938-948
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    • 2022
  • Gastric cancers (GC) are generally malignant tumors, occurring with high incidence and threatening public health around the world. Circular RNAs (circRNAs) play crucial roles in modulating various cancers, including GC. However, the functions of circRNAs and their regulatory mechanism in colorectal cancer (CRC) remain largely unknown. This study focuses on both the role of circCOL1A2 in CRC progression as well as its downstream molecular mechanism. Quantitative polymerase chain reaction (qPCR) and western blot were adopted for gene expression analysis. Functional experiments were performed to study the biological functions. Fluorescence in situ hybridization (FISH) and subcellular fraction assays were employed to detect the subcellular distribution. Luciferase reporter, RNA-binding protein immunoprecipitation (RIP), co-immunoprecipitation (Co-IP), RNA pull-down, and immunofluorescence (IF) and immunoprecipitation (IP) assays were used to explore the underlying mechanisms. Our results found circCOL1A2 to be not only upregulated in GC cells, but that it also propels the migration and invasion of GC cells. CircCOL1A2 functions as a competing endogenous RNA (ceRNA) by sequestering microRNA-1286 (miR-1286) to modulate ubiquitin-specific peptidase 10 (USP10), which in turn spurs the migration and invasion of GC cells by regulating RFC2. In sum, CircCOL1A2 sponges miR-1286 to promote cell invasion and migration of GC by elevating the expression of USP10 to downregulate the level of RFC2 ubiquitination. Our study offers a potential novel target for the early diagnosis and treatment of GC.

KIF26B-AS1 Regulates TLR4 and Activates the TLR4 Signaling Pathway to Promote Malignant Progression of Laryngeal Cancer

  • Li, Li;Han, Jiahui;Zhang, Shujia;Dong, Chunguang;Xiao, Xiang
    • Journal of Microbiology and Biotechnology
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    • 제32권10호
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    • pp.1344-1354
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    • 2022
  • Laryngeal cancer is one of the highest incidence, most prevalently diagnosed head and neck cancers, making it critically necessary to probe effective targets for laryngeal cancer treatment. Here, real-time quantitative reverse transcription PCR (qRT-PCR) and western blot analysis were used to detect gene expression levels in laryngeal cancer cell lines. Fluorescence in situ hybridization (FISH) and subcellular fractionation assays were used to detect the subcellular location. Functional assays encompassing Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), transwell and wound healing assays were performed to examine the effects of target genes on cell proliferation and migration in laryngeal cancer. The in vivo effects were proved by animal experiments. RNA-binding protein immunoprecipitation (RIP), RNA pulldown and luciferase reporter assays were used to investigate the underlying regulatory mechanisms. The results showed that KIF26B antisense RNA 1 (KIF26B-AS1) propels cell proliferation and migration in laryngeal cancer and regulates the toll-like receptor 4 (TLR4) signaling pathway. KIF26B-AS1 also recruits FUS to stabilize TLR4 mRNA, consequently activating the TLR4 signaling pathway. Furthermore, KIF26B-AS1 plays an oncogenic role in laryngeal cancer via upregulating TLR4 expression as well as the FUS/TLR4 pathway axis, findings which offer novel insight for targeted therapies in the treatment of laryngeal cancer patients.

Anaerobic Ammonium Oxidation Process in an Upflow Anaerobic Sludge Blanket Reactor with Granular Sludge Selected from an Anaerobic Digestor

  • Tran, Hung-Thuan;Park, Young-Joo;Cho, Mi-Kyeoung;Kim, Dong-Jin;Ahn, Dae-Hee
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제11권3호
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    • pp.199-204
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    • 2006
  • The purpose of this work was to evaluate the development of the anammox process by the use of granular sludge selected from a digestion reactor as a potential seed source in a lab-scale UASB (upflow anaerobic sludge blanket) reactor system. The reactor was operated for approximately 11 months and was fed by synthetic wastewater. After 200 days of feeding with $NH_4^+\;and\;NO_2^-$ as the main substrates, the biomass showed steady signs of ammonium consumption, resulting in over 60% of ammonium nitrogen removal. This report aims to present the results and to more closely examine what occurs after the onset of anammox activity, while the previous work described the start-up experiment and the presence of anammox bacteria in the enriched community using the fluorescence in situ hybridization (FISH) technique. By the last month of operation, the consumed $NO_2^--N/NH_4^+-N$ ratio in the UASB reactor was close to 1.32, the stoichiometric ratio of the anammox reaction. The obtained results from the influent-shutdown test suggested that nitrite concentration would be one key parameter that promotes the anammox reaction during the start-up enrichment of anammox bacteria from granular sludge. During the study period, the sludge color gradually changed from black to red-brownish.

병리검사를 위한 유방검체 고정의 질 향상 활동 (Quality Improvements of Breast Specimen Fixation for Histopathology Test)

  • 조승일;최명섭;배문환;이문정;우희숙;박순채;조은윤
    • 대한임상검사과학회지
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    • 제42권3호
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    • pp.149-154
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    • 2010
  • To improve accuracy of the immunohistochemical testing and fluorescence in situ hybridization (FISH) study as well as a routine histology diagnosis in breast cancer, quality improvement for optimal tissue handling is mandatory. We evaluate fraction defective of 7107 blocks from 349 breast cancer patients, who underwent surgical treatment at Samsung Medical Center Seoul, Korea from January 1, 2009 to March 31, 2010. We decided pre-improvement period from January, 2009 to June, 2009. In the first quality improvement period (July, 2009 to September, 2009) we made improvements in protocol of gross examination. In the second quality improvement period (October, 2009 to December, 2009) we attempted more effective formalin fixation such as frequent exchange of formalin and use of separate fixation container for each case. In the third quality improvement period (January, 2010 to March, 2010) improvement of tissue processor was performed. We achieved a marked reduction of fraction defective (9-16%) through efforts to improve quality of formalin-fixed, paraffin-embedded blocks when compared to pre-improvement period.

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Sperm DNA fragmentation and sex chromosome aneuploidy after swim-up versus density gradient centrifugation

  • Kim, Sung Woo;Jee, Byung Chul;Kim, Seul Ki;Kim, Seok Hyun
    • Clinical and Experimental Reproductive Medicine
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    • 제44권4호
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    • pp.201-206
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    • 2017
  • Objective: The aim of this study was to compare the efficacy of swim-up and density gradient centrifugation (DGC) for reducing the amount of sperm with fragmented DNA, sex chromosome aneuploidy, and abnormal chromatin structure. Methods: Semen samples were obtained from 18 healthy male partners who attended infertility clinics for infertility investigations and were processed with swim-up and DGC. The percentages of sperm cells with fragmented DNA measured by the sperm chromatin dispersion test, normal sex chromosomes assessed by fluorescence in situ hybridization, and abnormal chromatin structure identified by toluidine blue staining were examined. Results: The percentage of sperm cells with fragmented DNA was significantly lower in the swim-up fraction (9.7%, p= 0.001) than in the unprocessed fraction (27.0%), but not in the DGC fraction (27.8%, p= 0.098). The percentage of sperm cells with normal X or Y chromosomes was comparable in the three fractions. The percentage of sperm cells with abnormal chromatin structure significantly decreased after DGC (from 15.7% to 10.3%, p= 0.002). The swim-up method also tended to reduce the percentage of sperm cells with abnormal chromatin structure, but the difference was not significant (from 15.7% to 11.6%, p= 0.316). Conclusion: The swim-up method is superior for enriching genetically competent sperm.

한국 재래닭의 발생.발육단계별 telomere와 telomerase activity 분석

  • 정길선;조은정;최철환;손시환
    • 한국가금학회:학술대회논문집
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    • 한국가금학회 2004년도 제21차 정기총회 및 학술발표회
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    • pp.16-18
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    • 2004
  • 본 연구는 닭의 여러 조직별 세포들의 telomere 함유율과 telomerase 활성도를 분석 제시하고자 하였다. 한국재래계의 수정란 및 발생단계별 신생 조직과 출생 후 성장단계별 각 조직들에 대한 telomere의 함량과 telomerase 활성도를 분석하였고, 초기배자, 간, 뇌, 신장, 심장, 생식선 조직 및 백혈구 세포를 분석대상으로 하였다. Telomere의 함량 분석은 chicken telomeric DNA probe를 이용한 Q-FISH법으로 수행하였고, telomerase activity의 분석은 TRAP법을 이용하였다. 분석결과 초기 배자, 생식선 세포 및 신장세포에서는 지속적으로 매우 높은 telomerase activity를 나타내었으나 뇌, 심장, 간 등에서는 발생 및 발육이 진행됨에 따라 유의적 감소 양상을 보였다. 닭의 각 조직별 telomere의 함량 분석결과, 대부분의 세포들이 성장이 진행됨에 따라 telomere 함유율이 감소되는 양상을 보였다. 이상의 결과로부터 telomerase의 활성도와 telomere의 함량간에 매우 밀접한 연관성을 보이며. 이들이 닭 조직별 세포의 분화 및 증식성 특이성과도 밀접한 관련이 있는 것으로 나타났다.

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부분질산화와 혐기성 암모늄산화를 이용한 돈사폐수처리 (Piggery Waste Treatment using Partial Nitritation and Anaerobic Ammonium Oxidation)

  • 황인수;민경석;이영옥
    • 한국물환경학회지
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    • 제22권4호
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    • pp.599-604
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    • 2006
  • Nitrogen removal with the combined SHARON (Single reactor system for high ammonium removal over nitrite)ANAMMOX (Anaerobic ammonium oxidation) process using the effluent of ADEPT (Anaerobic digestion elutriated phased treatment) slurry reactor with very low C/N ratio for piggery waste treatment was investigated. For the preceding SHARON reactor, ammonium nitrogen loading and removal rate were $0.97kg\;NH_4-N/m^3_{reactor}/day$ and $0.68kg\;NH_4-N/m^3_{reactor}/day$ respectively. In steady state, bicarbonate alkalinity consumption for ammonium nitrogen converted to $NO_2-N$ or $NO_3-N$ was 8.4 gram per gram ammonium nitrogen. The successive ANAMMOX reactor was fed with the effluent from SHARON reactor. The loading and removal rate of the soluble nitrogen defined as the sum total of $NH_4-N$, $NO_2-N$ and $NO_3-N$ in ANAMMOX reactor were $1.36kg\;soluble\;N/m^3_{reactor}/day$ and $0.7kg\;soluble\;N/m^3_{reactor}/day$, respectively. The average $NO_2-N/NH_4-N$ removal ratio by ANAMMOX was 2.41. Fluorescence in situ hybridization (FISH) analysis verified that Candidatus Kuenenia stuttgartiensis were dominate, which means that they played an important role of nitrogen removal in ANAMMOX reactor.