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Chinese Journal of Cancer Research 2004年第4期16卷 260-264页

作者：王伟 Marianne Ifversen 赵春亭汪洪毅赵洪国 Department of Hematology Affiliated Hospital Qingdao University Medical College Qingdao 266003 China Bloodbank 2034 Laboratory Center Rigshospitalet Biegdamsvej 9 2100 Copenhagen Denmark

Objective: To establish a method to improve the detection of disseminated tumor cells in bone marrow and peripheral blood samples of neuroblastoma patients and analysis of cytogenetic aberration. Methods: Immunofluorescent staining was performed using a cocktail of primary monoclonal neuroblastoma antibodies (14.G2a, 5.1H11). fluorescence in situ hybridization was applied with fluorescent probes specific for Nmyc genes afterwards. A novel computer assisted scanning system for automatic search, image analysis and repositioning of these positive cells was developed. Fifty-six bone marrow and peripheral blood samples from 7 patients were evaluated by this method. Results: fluorescence in situ hybridization can be combined with immunofluorescent staining in detecting Nmyc amplification in neuroblastoma patients. fluorescence in situ hybridization results correlated well with data obtained by conventional cytogenetic procedures. Conclusion: The technique described allows search of tumor cells in the bone marrow as well as detection of Nmyc amplification in interphase nuclei.

关键词： Neuroblastoma fluorescence in situ hybridization Nmyc gene

Identification of Prorocentrum minimum and Takayama pulchella by fluorescence in situ hybridization through epifluorescence microscopy and flow cytometry

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Acta Oceanologica Sinica 2009年第2期28卷 103-114页

作者： HOU Jianjun LAI Hongyan HUANG Bangqin CHEN Jixin College of Fisheries Huazhong Agricultural University Wuhan 430070 China College of Life Sciences Hubei Normal University Huangshi 435002 China State Key Laboratory of Marine Environmental Science Environmental Science Research Center Xiamen University Xiamen 361005 China Key Laboratory of Freshwater Fish Germplasm Resources and Biotechnology Yangtse River Fisheries Research Institute Jinzhou 434000 China State Key Laboratory of Freshwater Ecology and Biotechnology Institute of Hydrobiology Chinese Academy of Sciences Wuhan 430072 China

Partial rDNA sequences of Prorocentrum minimum and Takayama pulchella were amplified, cloned and sequenced, and these sequence data were deposited in the GenBank. Eight oligonucleotide probes （DNA probes） were designed based on the sequence analysis. The probes were employed to detect and identify P. minimum and T. pulchella in unialgal and mixed algal samples with a fluorescence in situ hybridization method using flow cytometry. Epifluorescence micrographs showed that these specific probes labeled with fluorescein isothiocyanate entered the algal cells and bound to target sequences, and the fluorescence signal resulting from whole-cell hybridization varied from probe to probe. These DNA probes and the hybridization protocol we developed were specific and effective for P. minimum and T. pulchella, without any specific binding to other algal species. The hybridization efficiency of different probes specific to P. minimum was in the order： PM18S02 PM28S02 〉 PM28S01 〉PM18S01, and that of the probes specific to T. pulcheUa was TP18S02 TP28S01 〉 TP28S02 〉TP18S01. The different hybridization efficiency of the DNA probes could also be shown in the fluorescent signals between the labeled and unlabeled cells demonstrated using flow cytometry. The DNA probes PM18S02, PM28S02, TP18S02 and TP28S01, and the protocol, were also useful for the detection of Mgae in natural samples.

关键词： oligonucleotide DNA probes Prorocentrum minimum Takayama pulcheUa fluorescence in situ hybridization flow cytometry

Niche Differentiation of Phenol-Degrading Microorganisms in UASB Granular Sludge as Revealed by fluorescence in situ hybridization

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Engineering 2022年第2期8卷 61-66页

作者： Kengo Kubota Kei Igarashi Masayoshi Yamada Yasuyuki Takemura Yu-You Li Hideki Harada Department of Civil and Environmental Engineering Tohoku UniversitySendai 980-8579Japan Department of Urban Environmental Design and Engineering National Institute of TechnologyKagoshima CollegeKirishima 899-5193Japan New Industry Creation Hatchery Center Tohoku UniversitySendai 980-8579Japan

A microbial community structure of granules harvested from an anaerobic sludge blanket reactor treating phenolic wastewater was investigated using fluorescence in situ hybridization(FISH)and clone library *** of Syntrophorhabdaceae and Cryptanaerobacter were observed to be responsible for phenol *** accurate taxonomic assignment of Cryptanaerobacter clones,phylogenetic analysis using nearly full-length 16S ribosomal RNA(rRNA)gene sequences was *** oligonucleotide probes were designed to detect the following three taxonomic groups:Syntrophorhabdaceae,Cryptanaerobacter,and *** analysis of thin sections of anaerobic granules showed a random distribution of bacteria and ***,a well-defined distribution of Syntrophorhabdaceae,Cryptanaerobacter,and Syntrophus was *** and Syntrophus were found on the outer layer of the granules and were closely associated with each other,while Syntrophorhabdaceae was located in the deeper part of the *** specific distribution of the bacteria is most likely due to their metabolic association and affinity for the *** degradation in the granular sludge was observed to be carried out in the following ***,Cryptanaerobacter converts phenol to benzoate,which is then degraded by Syntrophus into *** syntrophic degradation of phenol occurs near the surface of the granule,where the phenol concen-tration is *** the deeper part of the granule,where the phenol concentration is lower,Syntrophorhabdaceae degrades phenol into *** observed that Syntrophorhabdaceae is less likely to produce benzoate as an intermediate to feed the neighboring organisms,which contradicts the theo-ries presented by previous studies.

关键词： Cryptanaerobacter fluorescence in situ hybridization Anaerobic phenol degradation Syntrophorhabdaceae Syntrophus UASB granular sludge

Development of taxonomic rRNA-targeted probes of two harmful algae: Prorocentrum minimum and Karenia mikimotoi by fluorescence in situ hybridization

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Acta Oceanologica Sinica 2013年第2期32卷 66-75页

作者： CHEN Guofu WANG Quanfu ZHANG Chunyun ZHANG Baoyu WANG Guangce LU Douding XU Zhong YAN Peishen Harbin Institute of Technology (Weihai) The First Institute of Oceanography State Ocenic Administration Institute of Oceanology Chinese Academy of Sciences The Second Institute of Oceanography State Ocenic Administration

Harmful algal blooms recently have been under the spotlight throughout the world, because of their nega- tive impact on the marine environment, aquaculture, fisheries as well as public health. The development of methods for rapid and precise identification and quantification of causative species is essential for the warning and monitoring of blooms, among which the techniques based on taxonomic probes are the most favored. In this study, two harmful algae, i.e., Prorocentrum minimum and Karenia mikimotoi were tak- en into consideration. The partial large subunit rDNA （D1-D2） of both species were firstly PCR-amplified, cloned and sequenced. The obtained sequences were then introduced to carry out alignment analysis for gene specific regions. Three respective candidate probes for each species were designed and used to screen the optimal probe by performing fluorescence in situ hybridization （FISH） tests. The results showed that the probes Pmin0443 and Kmik0602 displayed the best hybridization for P. minimum and K.. mikimotoi, respectively. Both the specific （taxonomic）（Pmin0443 and Kmik0602） and the control probes （UniC0512 and UniR0499） were used for cross-reactivity tests with other microalgae in our laboratory. The probes Pmin0443 and Kmik0602 are specific and could be served as taxonomic probes introduced into the tech- niques targeting rRNA, such as FISH, sandwich hybridization, and DNA-microarray assay of P minimum and K. mikimotoi in the future. Finally, FISH analyses with both probes were performed on the simulated field samples. The probes could hybridize exclusively with the target cells well, and no significant differ- ence （p 〉0.05） was observed in the cell densities of the samples determined by FISH and light microscopy （LM）. All suggest that the probes are specific and could be introduced into FISH for the monitoring of both harmful algae.

关键词： Prorocentrum minimum Karenia mikimotoi fluorescence in situ hybridization taxonomic probe

Optimizing the hybridization chain reaction-fluorescence in situ hybridization(HCR-FISH)protocol for detection of microbes in sediments

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Marine Life Science & Technology 2021年第4期3卷 529-541页

作者： Zeyu Jia Yijing Dong Heng Xu Fengping Wang State Key Laboratory of Microbial Metabolism Joint International Research Laboratory of Metabolic and Developmental SciencesSchool of Life Sciences and BiotechnologyShanghai Jiao Tong UniversityShanghai 200240China School of Physics and Astronomy Shanghai Jiao Tong UniversityShanghai 200240China Institute of Natural Science Shanghai Jiao Tong UniversityShanghai 200240China School of Oceanography Shanghai Jiao Tong UniversityShanghai 200240China

fluorescence in situ hybridization(FISH)is a canonical tool commonly used in environmental microbiology research to visualize targeted ***,the problems of low signal intensity and false-positive signals impede its widespread ***,the signal intensity can be amplified by incorporating hybridization Chain Reaction(HCR)with FISH,while the specificity can be improved through protocol modification and proper *** we optimized the HCR-FISH protocol for studying microbes in environmental samples,particularly marine ***,five sets of HCR initiator/amplifier pairs were tested on the laboratory-cultured bacterium Escherichia coli and the archaeon Methano-coccoides methylutens,and two sets displayed high hybridization efficiency and ***,we tried to find the best combination of sample pretreatment methods and HCR-FISH protocol for environmental sample analysis with the aim of producing less false positive *** detachment methods,extraction methods and formulas of hybridization buffer were tested using sediment ***,an image processing method was developed to enhance the DAPI signal of microbial cells against that of abiotic particles,providing a reliable reference for FISH *** summary,our optimized HCR-FISH protocol showed promise to serve as an addendum to traditional FISH for research on environmental microbes.

关键词： fluorescence in situ hybridization hybridization chain reaction HCR-FISH Microbial detection Sediment

A populational survey of 45S rDNA polymorphism in the Jefferson salamander Ambystoma jeffersonianum revealed by fluorescence in situ hybridization(FISH)

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Current Zoology 2009年第2期55卷 145-149页

作者： James P. BOGART Department of Integrative Biology University of GuelphGuelphOntario N1G 2W1Canada

The chromosomal localization of 45S ribosomal RNA genes in Ambystoma jeffersonianum was determined by fluorescence in situ hybridization with 18S rDNA fragment as a probe (FISH-rDNA). Our results revealed the presence of rDNA polymorphism among *** populations in terms of number,location and FISH signal intensity on the chromosomes. Nine rDNA cytotypes were found in ten geographically isolated populations and most of them contained derivative rDNA sites. Our preliminary study provides strong indication of karyotypic diversification of *** that is demonstrated by intraspecific variation of 45S rDNA cytotypes. rDNA cytotype polymorphism has been described in many other caudate amphibians. We predict that habitat isolation,low dispersal ability and decline of effective population size could facilitate the fixation and accumulation of variable rDNA cytotypes during their chromosome evolution.

关键词： rDNA Polymorphism Jefferson salamander Ambystoma jeffersonianum fluorescence in situ hybridization FISH-rDNA

Association of chromosome 7 aneuploidy measured by fluorescence in situ hybridization assay with muscular invasion in bladder cancer

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Cancer Communications 2020年第4期40卷 167-180页

作者： Xiayao Diao Jinhua Cai Junjiong Zheng Jianqiu Kong Shaoxu Wu Hao Yu Hao Huang Weibin Xie Xiong Chen Chengran Huang Lifang Huang Haide Qin Jian Huang Tianxin Lin Department of Urology Sun Yat-sen Memorial HospitalSun Yat-senUniversityGuangzhouGuangdong510120P.R.China Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation Sun Yat-Sen Memorial HospitalSun Yat-Sen UniversityGuangzhouGuangdong510120P.R.China Department of Neurology Sun Yat-sen Memorial HospitalSun Yat-sen UniversityGuangzhouGuangdong510120P.R.China State Key Laboratory of Oncology in South China GuangzhouGuangdong510120P.R.China

Background:The preoperative prediction of muscular invasion status is important for adequately treating bladder cancer(BC)but nevertheless,there are some existing dilemmas in the current preoperative diagnostic accuracy of BC with muscular ***,we investigated the potential association between the fluorescence in situ hybridization(FISH)assay and muscular invasion among patients with BC.A cytogenetic-clinical nomogram for the individualized preoperative differentiation of muscle-invasive BC(MIBC)from non-muscle-invasive BC(NMIBC)is also ***:All eligible BC patients were preoperatively tested using a FISH assay,which included 4 sites(chromosome-specific centromeric probe[CSP]3,7,and 17,and gene locus-specific probe[GLP]-p16 locus).The correlation between the FISH assay and BC muscular invasion was evaluated using the Chi-square *** the training set,univariate and multivariate logistic regression analyses were used to develop a cytogenetic-clinical nomogram for preoperative muscular invasion ***,we assessed the performance of the nomogram in the training set with respect to its discriminatory accuracy and calibration for predicting muscular invasion,and clinica usefulness,which were then validated in the validation ***,model comparison was set to evaluate the discrimination and clinical usefulness between the nomogram and the individual variables incorporated in the ***:Muscular invasion was more prevalent in BC patients with positive CSP3,CSP7 and CSP17 status(OR[95%CI],2.724[1.555 to 4.774],P<0.001;3.406[1.912 to 6.068],P<0.001 and 2.483[1.436 to 4.292],P=0.001,respectively).Radiologydetermined tumor size,radiology-determined clinical tumor stage and CSP7 status were identified as independent risk factors of BC muscular invasion by the multivariate regression analysis in the training ***,a cytogenetic-clinical nomogram incorporating these three independent risk factors was constructed and was observed

关键词： chromosome aneuploidy fluorescence in situ hybridization muscular invasion nomogram preoperative prediction urinary bladder neoplasms

fluorescence in situ hybridization polymorphism usinng 45S and 5S rDNA in different cultivars of sweet potato

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Fluorescence in situ hybridization polymorphism usinng 45S a...

Application of fluorescence in situ hybridization in the detection of bladder transitional-cell carcinoma: A multi-center clinical study based on Chinese population

江苏省遗传学会2016年学术年会

作者： Jianying Sun Yonghua Han Zongyun Li Institute of Integrative Plant Biology School of Life ScienceJiangsu Normal University

To detect if polymorphism could be observed in the hybridization patterns of different sweet potato cultivars,twenty sweet potato cultivars were analysed with two-colour fluorescence in situ hybridization（FISH） using the 45 S rDNA and 5 S *** number、region and intensity of rDNA loci on the metaphase chromosmes and interphase nucleus of all materials were *** average number of chromosmes in these cultivars is 90,but not always be.19 to 22 45 S rDNA signals should be examined,varies among *** were 3 regions of 45 S rDNA hybridization signals located in metaphase *** 45 S rDNA signals were detected on the ends of chromosomes with high intensity brightness,weaker signals on the satellites and the weakest on the *** intensity of 45 S rDNA differs in different *** comparing the most strong signals,5 cultivars on the chromosome long arm,2 cultivars covering the entire chromosome and 2 cultivars on the short *** number of 5 S rDNA was consistent among these cultivars,all of them have 6 signals with different *** location of 5 S rDNA is usually on the centromeric and the middle of chromosome *** former generally have a stronger signal,the latter have the *** both of the 45 S and 5 S rDNA are present,their signals will always be weak,and they usually located near the pericentromere and in the middle of chromosme *** this study,a low level of polymorphism has been observed in the FISH pattern of various types of sweet potato *** results may provide additional information on the influence of different origin on sweet potato genetics stocks.

关键词： fluorescence in situ hybridization sweet potato rDNA

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Asian Journal of Urology 2019年第1期6卷 114-121页

作者： Liqun Zhou Kaiwei Yang Xuesong Li Yi Ding Dawei Mu Hanzhong Li Yong Yan Jinyi Li Dongwen Wang Wei Li Yulong Cong Jiangping Gao Kewei Ma Yajun Xiao Sheng Zhang Hongyi Jiang Weilie Hu Qiang Wei Xunbo Jin Zhichen Guan Qingyong Liu Danfeng Xu Xin Gao Yongguang Jiang Weimin Gan Guang Sun Qing Wang Yanhui Liu Jianquan Hou Liping Xie Xishuang Song Fengshuo Jin Jiafu Feng Ming Cai Zhaozhao Liang Jie Zhang Dingwei Ye Lin Qi Lulin Ma Jianzhong Shou Yuping Dai Jianyong Shao Ye Tian Shizhe Hong Tao Xu Chuize Kong Zefeng Kang Yuexin Liu Xun Qu Benkang Shi Shaobin Zheng Yi Lin Shujie Xia Dong Wei Jianbo Wu Weiling Fu Zhiping Wang Jianbo Liang Department of Urology Peking University First HospitalBeijingChina Institute of Urology Peking UniversityBeijingChina National Urological Cancer Center BeijingChina Air Force General Hospital PLABeijingChina Department of Urology Peking Union Medical College HospitalBeijingChina Department of Urology Beijing Chao-Yang HospitalBeijingChina Department of Urology Armed-Police General HospitalBeijingChina Department of Urology Shanxi Medical University First HospitalTaiyuanChina Department of Urology Hebei Medical University Second HospitalShijiazhuangChina Department of Urology The General Hospital of the People’s Liberation ArmyBeijingChina Department of Urology First Affiliated Hospital of Jilin UniversityChangchunChina Department of Urology Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina Department of Pathology The First Affiliated Hospital of Fujian Medical UniversityFuzhouChina Department of Urology The Second Xiangya Hospital of Central South UniversityChangshaChina Department of Urology General Hospital of Guangzhou Military Command of PLAGuangzhouChina Department of Urology West China School of MedicineChengduChina Department of Urology Shangdong Province-owned HospitalJinanChina Department of Urology Peking University Shenzhen HospitalShenzhenChina Department of Urology The Central Hospital of Jinan CityJinanChina Department of Urology Shanghai Changzheng HospitalShanghaiChina Department of Urology The Third Affiliated Hospital of Sun Yat-sen UniversityGuangzhouChina Department of Urology Beijing Anzhen Hospital of Capital Medical UniversityBeijingChina Department of Urology The Second Affiliated Hospital of Xi’an Jiaotong UniversityXi’anChina Department of Urology The Second Hospital of Tianjin Medical UniversityTianjinChina Department of Urology People’s Hospital of Xinjiang Uygur Autonomous RegionWulumuqiChina Department of Urology Guangdong General HospitalGuangzhouChina Department of Urology The First Affil

Objective:To evaluate the diagnostic value of fluorescence in situ hybridization(FISH)in bladder ***:We enrolled healthy volunteers and patients who were clinically suspected to have bladder cancer and conducted FISH tests and cytology examinations from August 2007 to December *** operating characteristic(ROC)curve analysis was performed and the area under curve(AUC)values were calculated for both the FISH and urine cytology ***:A cohort of 988 healthy volunteers was enrolled to establish a reference range for the normal population.A total of 4807 patients with hematuria were prospectively,randomly enrolled for the simultaneous analysis of urine cytology,FISH testing,and a final diagnosis as determined by the pathologic findings of a biopsy or a surgically-excised ***,the sensitivity of FISH in detecting transitional-cell carcinoma was 82.7%,while that of cytology was 33.4%(p<0.001).The sensitivity values of FISH for non-muscle invasive and muscle invasive bladder transitional-cell carcinoma were 81.7%and 89.6%,respectively(p=0.004).The sensitivity values of FISH for low and high grade bladder cancer were 82.6%and 90.1%,respectively(p=0.002).Conclusion:FISH is significantly more sensitive than voided urine cytology for detecting bladder cancer in patients evaluated for gross hematuria at all cancer grades and *** sensitivity using FISH was obtained in high grade and muscle invasive tumors.

关键词： Bladder transitionalcell carcinoma fluorescence in situ hybridization Detection Grade Stage

Karyotyping of Brassica oleracea *** on rDNA and Cot-1 DNA fluorescence in situ hybridization

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Frontiers in Biology 2007年第4期2卷 403-407页

作者： WANG Taixia WU Chunhong HUANG Jinyong WEI Wenhui College of Life Sciences Henan Normal UniversityXinxiang 453007China Library Henan Polytechnic UniversityJiaozuo 454000China College of Life Sciences Zhengzhou UniversityZhengzhou 450001China Institute of Oil Crops Chinese Academy of Agricultural SciencesWuhan 430062China

To explore an effective and reliable karyotyping method in Brassica crop plants,Cot-1 DNA was isolated from Brassica oleracea genome,labeled as probe with Biotin-Nick Translation Mix kit,in situ hybridized to mitotic spreads,and where specific fluorescent bands showed on each chromosome pair.25S and 5S rDNA were labeled as probes with DIG-Nick Translation Mix kit and Biotin-Nick Translation Mix kit,respectively,in situ hybridized to mitotic preparations,where 25S rDNA could be detected on two chromosome pairs and 5S rDNA on only ***-1 DNA contains rDNA and chromosome sites identity between Cot-1 DNA and 25S rDNA was determined by dual-colour fluores-cence in situ *** these showed that the karyo-typing technique based on a combination of rDNA and Cot-1 DNA chromosome landmarks is superior to all but one.A more exact karyotype of *** has been analyzed based on a combination of rDNA sites,Cot-1 DNA fluorescent bands,chromosome lengths and arm ratios.

关键词： Brassica oleracea L. rDNA Cot-1 DNA fluorescence in situ hybridization karyotyping