PCR enzyme linked immune sorbent assay (ELISA) was developed for detection of RT PCR amplified cytokeratin 19 (CK19) mRNA and carcinoembryonic antigen (CEA) mRNA. The non radioactive hybridization was performed in ...PCR enzyme linked immune sorbent assay (ELISA) was developed for detection of RT PCR amplified cytokeratin 19 (CK19) mRNA and carcinoembryonic antigen (CEA) mRNA. The non radioactive hybridization was performed in a streptavidin coated microwell with digoxigenin labeled PCR products and with biotin labeled capture probe. PCR ELISA was proved to be expedient, simple, sensitive and safe for identification of CK19 , CEA RT PCR products. These results were proven by sequencing.展开更多
Embryo abortion stage and rescue system of hybrids were studied in the distant hybridizationbetween plum and apricot. Identification of the hybrids was also made. The resultsshowed: (1) Embryo abortion started from th...Embryo abortion stage and rescue system of hybrids were studied in the distant hybridizationbetween plum and apricot. Identification of the hybrids was also made. The resultsshowed: (1) Embryo abortion started from three weeks after pollination. (2) The germinationand growth of embryos were different at different growth stages, which could germinateand grow with PF value>0.5, but failed with PF value<0.5. In embryo rescue system ofhybrids, the best germination and differentiation medium was MS+6-BA 2mgL-1+IAA 0.3mgL-1,the rate of germination and differentiation reached up to 80%, bud induction andmultiplication medium was MS+6-BA 1.5mgL-1+IAA 0.3mgL-1, rooting medium was 1/2 MS+IAA0.8mgL-1. Some hybrids were transplanted into the field successfully. (3) Leaf shapeinvestigation and identification by S allele-specific PCR and RAPDs showed that thehybrids were true ones.展开更多
Dot-blot hybridization was applied in this paper to detect five viruses infecting cucurbitaceous crops, Zuccini yellow mosaic virus （ZYMV）, Watermelon mosaic virus （WMV）, Cucumber mosaic virus （CMV）, Papaya ring...Dot-blot hybridization was applied in this paper to detect five viruses infecting cucurbitaceous crops, Zuccini yellow mosaic virus （ZYMV）, Watermelon mosaic virus （WMV）, Cucumber mosaic virus （CMV）, Papaya ringspot viruswatermelon strain （PRSV-W） and Squash mosaic virus （SqMV）, as a good alternative assay in seed health test and epidemiological and transgenic research. Digoxigenin-labelled cDNA probes of the five viruses were synthesized by PCR with the specific primers and applied in dot-blot hybridization to detect five viruses in crude extraction of the infected leaves. And three SqMV probes of different lengths （0.55, 1.6, and 2.7 kb, respectively） were designed to investigate the effect of hybridization. The results showed that the sensitivity for detecting the crude extraction of infected leaves by ZYMV, WMV, CMV, PRSV-W, and SqMV was down to 1：160, 1：160, 1：320, 1：160, and 1：320, respectively. Three SqMV probes of different length showed no differences on the sensitivity and specificity. The digoxigenin-labelled probes prepared by PCR could be used for accurate and rapid identification of 5 viruses infecting cucurbitaceous crops with good stabilities, sensitivities, specificity, and reproducibilifies.展开更多
Chromosome aberrations are distinctive features of human malignant tumors. Analysis of chromosomal changes can illuminate the molecular mechanisms underlying the development and progression of cancer. To establish the...Chromosome aberrations are distinctive features of human malignant tumors. Analysis of chromosomal changes can illuminate the molecular mechanisms underlying the development and progression of cancer. To establish the technique of multicolor fluorescence in situ hybridization （M-FISH） for identifying chromosome aberrations in esophageal carcinoma cell line KYSE 410-4, four pools of 6-color whole-chromosome painting probes have been designed and hybridized on the same metaphase spread by four rounds of repetitive FISH. Repetitive 6-color M-FISH was successfully established and the cytogenetic abnormalities in KYSE 410-4 cells were characterized. Chromosome gains occurred at 2q, 3, 8, 17p, and X. An isochromosome 3q was visualized in the cell line, which might be one intermediate mechanism leading to 3p losses and/or 3q gains. Furthermore, 16 structural arrangements were detected, including four derivative chromosomes. The rearrangement of the centromeric regions accounted for approximately 44% of all rearrangements. The results added a more complete and accurate information of the genetic alterations to the classical cytogenetic description of KYSE 410-4 and provided a detailed cytogenetic background data for appropriate use of the cell line. The established 6-color M-FISH was useful for analyzing chromosomes in the whole genome of human tumors.展开更多
A set of novel structural descriptors (molecular hybridization electronegativity-distance vector, VMEDh) was put forward, and the quantitative structure–activity relationship (QSAR) of a series of 17α-Acetoxyprogest...A set of novel structural descriptors (molecular hybridization electronegativity-distance vector, VMEDh) was put forward, and the quantitative structure–activity relationship (QSAR) of a series of 17α-Acetoxyprogesterones (APs) was investigated. Taking into account the effect of various hybridized orbits on atomic electronegativities, we developed the structure descriptors with amended electronegativities to build a QSAR model. The 10-parameter model based on VMEDh yields a correlation coefficient R=0.972 and standard deviation SD=0.262, which are more desirable than those of the previous molecular electonegativity-distance vector (MEDV-4) (R=0.969, SD=0.275). By stepwise multiple linear regression, several parameters are selected to construct optimal models. The 7-parameter model based on VMEDh has R=0.960 and SD=0.276; its correlation coefficient (RCV) and standard deviation (SDCV) for leave-one-out procedure crossvalidation are respectively RCV=0.890 and SDCV=0.445. The 6-parameter MEDV-4 model has R=0.946, SD=0.304, RCV=0.903 and SDCV=0.406. It is demonstrated that VMEDh has desirable estimation performance and good predictive capability for this series of chemical compounds.展开更多
文摘PCR enzyme linked immune sorbent assay (ELISA) was developed for detection of RT PCR amplified cytokeratin 19 (CK19) mRNA and carcinoembryonic antigen (CEA) mRNA. The non radioactive hybridization was performed in a streptavidin coated microwell with digoxigenin labeled PCR products and with biotin labeled capture probe. PCR ELISA was proved to be expedient, simple, sensitive and safe for identification of CK19 , CEA RT PCR products. These results were proven by sequencing.
基金supported by the Technological Production Transformation Foundation by the Ministry of Science and Technology of China(2002370010495)the foundation of Shandong Fruit Tree Three-Zero Project
文摘Embryo abortion stage and rescue system of hybrids were studied in the distant hybridizationbetween plum and apricot. Identification of the hybrids was also made. The resultsshowed: (1) Embryo abortion started from three weeks after pollination. (2) The germinationand growth of embryos were different at different growth stages, which could germinateand grow with PF value>0.5, but failed with PF value<0.5. In embryo rescue system ofhybrids, the best germination and differentiation medium was MS+6-BA 2mgL-1+IAA 0.3mgL-1,the rate of germination and differentiation reached up to 80%, bud induction andmultiplication medium was MS+6-BA 1.5mgL-1+IAA 0.3mgL-1, rooting medium was 1/2 MS+IAA0.8mgL-1. Some hybrids were transplanted into the field successfully. (3) Leaf shapeinvestigation and identification by S allele-specific PCR and RAPDs showed that thehybrids were true ones.
文摘Dot-blot hybridization was applied in this paper to detect five viruses infecting cucurbitaceous crops, Zuccini yellow mosaic virus （ZYMV）, Watermelon mosaic virus （WMV）, Cucumber mosaic virus （CMV）, Papaya ringspot viruswatermelon strain （PRSV-W） and Squash mosaic virus （SqMV）, as a good alternative assay in seed health test and epidemiological and transgenic research. Digoxigenin-labelled cDNA probes of the five viruses were synthesized by PCR with the specific primers and applied in dot-blot hybridization to detect five viruses in crude extraction of the infected leaves. And three SqMV probes of different lengths （0.55, 1.6, and 2.7 kb, respectively） were designed to investigate the effect of hybridization. The results showed that the sensitivity for detecting the crude extraction of infected leaves by ZYMV, WMV, CMV, PRSV-W, and SqMV was down to 1：160, 1：160, 1：320, 1：160, and 1：320, respectively. Three SqMV probes of different length showed no differences on the sensitivity and specificity. The digoxigenin-labelled probes prepared by PCR could be used for accurate and rapid identification of 5 viruses infecting cucurbitaceous crops with good stabilities, sensitivities, specificity, and reproducibilifies.
基金Acknowledgements This study was supported by the National Science Foundation （No. 30630067）；the State Key Basic Research Grant of China （No. 2004CB518705）； the Program for Changjiang Scholars and Innovative Research Team in University （No. IRT0416）.
文摘Chromosome aberrations are distinctive features of human malignant tumors. Analysis of chromosomal changes can illuminate the molecular mechanisms underlying the development and progression of cancer. To establish the technique of multicolor fluorescence in situ hybridization （M-FISH） for identifying chromosome aberrations in esophageal carcinoma cell line KYSE 410-4, four pools of 6-color whole-chromosome painting probes have been designed and hybridized on the same metaphase spread by four rounds of repetitive FISH. Repetitive 6-color M-FISH was successfully established and the cytogenetic abnormalities in KYSE 410-4 cells were characterized. Chromosome gains occurred at 2q, 3, 8, 17p, and X. An isochromosome 3q was visualized in the cell line, which might be one intermediate mechanism leading to 3p losses and/or 3q gains. Furthermore, 16 structural arrangements were detected, including four derivative chromosomes. The rearrangement of the centromeric regions accounted for approximately 44% of all rearrangements. The results added a more complete and accurate information of the genetic alterations to the classical cytogenetic description of KYSE 410-4 and provided a detailed cytogenetic background data for appropriate use of the cell line. The established 6-color M-FISH was useful for analyzing chromosomes in the whole genome of human tumors.
基金Funded by Chongqing Medical University Scientific Research Foundation
文摘A set of novel structural descriptors (molecular hybridization electronegativity-distance vector, VMEDh) was put forward, and the quantitative structure–activity relationship (QSAR) of a series of 17α-Acetoxyprogesterones (APs) was investigated. Taking into account the effect of various hybridized orbits on atomic electronegativities, we developed the structure descriptors with amended electronegativities to build a QSAR model. The 10-parameter model based on VMEDh yields a correlation coefficient R=0.972 and standard deviation SD=0.262, which are more desirable than those of the previous molecular electonegativity-distance vector (MEDV-4) (R=0.969, SD=0.275). By stepwise multiple linear regression, several parameters are selected to construct optimal models. The 7-parameter model based on VMEDh has R=0.960 and SD=0.276; its correlation coefficient (RCV) and standard deviation (SDCV) for leave-one-out procedure crossvalidation are respectively RCV=0.890 and SDCV=0.445. The 6-parameter MEDV-4 model has R=0.946, SD=0.304, RCV=0.903 and SDCV=0.406. It is demonstrated that VMEDh has desirable estimation performance and good predictive capability for this series of chemical compounds.
