BACKGROUND Genetic tests are increasingly performed for the management of unresectable pancreatic cancer.For genotyping aimed samples current guidelines recommend using core specimens,although based on moderate qualit...BACKGROUND Genetic tests are increasingly performed for the management of unresectable pancreatic cancer.For genotyping aimed samples current guidelines recommend using core specimens,although based on moderate quality evidence.However,in clinical practice among the endoscopic ultrasound(EUS) guided tissue acquisition methods,fine needle aspiration(FNA) is the most widely performed.AIM To assess the adequacy for next generation sequencing(NGS) of the DNA yielded from EUS-FNA pancreatic adenocarcinoma(PDAC) samples.METHODS Between November 2018 and December 2021,105 patients with PDAC confirmed by EUS-FNA were included in the study at our tertiary gastroenterology center.Either 22 gauge(G) or 19G FNA needles were used.One pass was dedicated to DNA extraction.DNA concentration and purity(A260/280,A260/230) were assessed by spectrophotometry.We assessed the differences in DNA parameters according to needle size and tumor characteristics(size,location) and the adequacy of the extracted DNA for NGS(defined as A260/280 ≥ 1.7,and DNA yield:≥ 10 ng for amplicon based NGS,≥ 50 ng for whole exome sequencing [WES],≥ 100 ng for whole genome sequencing [WGS]) by analysis of variance and ttest respectively.Moreover,we compared DNA purity parameters across the different DNA yield categories.RESULTS Our cohort included 49% male patients,aged 67.02 ± 8.38 years.The 22G needle was used in 71%of the cases.The DNA parameters across our samples varied as follows:DNA yield:1289 ng(inter quartile range:534.75-3101),A260/280 = 1.85(1.79-1.86),A260/230 = 2.2(1.72-2.36).DNA yield was > 10 ng in all samples and > 100 ng in 93% of them(one sample < 50 ng).There were no significant differences in the concentration and A260/280 between samples by needle size.Needle size was the only independent predictor of A260/230 which was higher in the 22G samples(P =0.038).NGS adequacy rate was 90% for 19G samples regardless of NGS type,and for 22G samples it reached 89% for WGS adequacy and 91% for WES and amplicon based NGS.Samples with DNA yield > 100 ng had significantly higher A260/280(1.89 ± 0.32 vs 1.34 ± 0.42,P = 0.013).Tumor characteristics were not corelated with the DNA parameters.CONCLUSION EUS-FNA PDAC samples yield DNA adequate for subsequent NGS.DNA amount was similar between 22G and 19G FNA needles.DNA purity parameters may vary indirectly with needle size.展开更多
Silicon phthalocyanine derivatives 1a and 1b were synthesized and characterized by UV, <sup>1</sup>H-NMR and MS. The photophysical properties of the compounds in DMSO were investigated. The maximum absorpt...Silicon phthalocyanine derivatives 1a and 1b were synthesized and characterized by UV, <sup>1</sup>H-NMR and MS. The photophysical properties of the compounds in DMSO were investigated. The maximum absorption peaks of compounds 1a and 1b at the Q-band are 681 nm. With ZnPc (Φ<sub>F</sub> = 0.20, Φ<sub>Δ</sub> = 0.67) as a reference, the fluorescence quantum yield (Φ<sub>F</sub>) of 1a and 1b are 0.20 and 0.31 respectively, and the singlet oxygen quantum yield (Φ<sub>Δ</sub>) are 0.66 and 0.59 respectively. The DNA-photocleavage activities of compounds 1a and 1b were studied by gel electrophoresis. Compounds 1a and 1b possess good photocleavage activity to pBR322 DNA. The results demonstrate that compounds 1a and 1b are potential photosensitizers for tumor therapy.展开更多
Telomeres are protein-DNA complexes at the terminals of linear chromosomes, which protect chromoso-mal integrity and maintain cellular replicative capacity. From single-cell organisms to advanced animals and plants, s...Telomeres are protein-DNA complexes at the terminals of linear chromosomes, which protect chromoso-mal integrity and maintain cellular replicative capacity. From single-cell organisms to advanced animals and plants, structures and functions of telomeres are both very conser-vative. In cells of human and vertebral animals, telomeric DNA base sequences all are (TTAGGG)n. In the present work, we have obtained absorption and fluorescence spectra measured from seven synthesized oligonucleotides to simu-late the telomeric DNA system and calculated their relative fluorescence quantum yields on which not only telomeric DNA characteristics are predicted but also possibly the shortened telomeric sequences during cell division are im-plied. Oligonucleotide 5’- TTAGGGTTAGGG holds a low relative fluorescence quantum yield and remarkable excitation energy innerconversion, which tallies with the telomeric sequence of (TTAGGG)n. This result shows that telomeric DNA has a strong non-radiative or innerconvertible capabil-展开更多
基金The Executive Agency for Higher Education,Research,Development and Innovation Funding-research,No.PN-Ⅲ-P1-1.2-PCCDI-2017-0797 (PANCNGS)
文摘BACKGROUND Genetic tests are increasingly performed for the management of unresectable pancreatic cancer.For genotyping aimed samples current guidelines recommend using core specimens,although based on moderate quality evidence.However,in clinical practice among the endoscopic ultrasound(EUS) guided tissue acquisition methods,fine needle aspiration(FNA) is the most widely performed.AIM To assess the adequacy for next generation sequencing(NGS) of the DNA yielded from EUS-FNA pancreatic adenocarcinoma(PDAC) samples.METHODS Between November 2018 and December 2021,105 patients with PDAC confirmed by EUS-FNA were included in the study at our tertiary gastroenterology center.Either 22 gauge(G) or 19G FNA needles were used.One pass was dedicated to DNA extraction.DNA concentration and purity(A260/280,A260/230) were assessed by spectrophotometry.We assessed the differences in DNA parameters according to needle size and tumor characteristics(size,location) and the adequacy of the extracted DNA for NGS(defined as A260/280 ≥ 1.7,and DNA yield:≥ 10 ng for amplicon based NGS,≥ 50 ng for whole exome sequencing [WES],≥ 100 ng for whole genome sequencing [WGS]) by analysis of variance and ttest respectively.Moreover,we compared DNA purity parameters across the different DNA yield categories.RESULTS Our cohort included 49% male patients,aged 67.02 ± 8.38 years.The 22G needle was used in 71%of the cases.The DNA parameters across our samples varied as follows:DNA yield:1289 ng(inter quartile range:534.75-3101),A260/280 = 1.85(1.79-1.86),A260/230 = 2.2(1.72-2.36).DNA yield was > 10 ng in all samples and > 100 ng in 93% of them(one sample < 50 ng).There were no significant differences in the concentration and A260/280 between samples by needle size.Needle size was the only independent predictor of A260/230 which was higher in the 22G samples(P =0.038).NGS adequacy rate was 90% for 19G samples regardless of NGS type,and for 22G samples it reached 89% for WGS adequacy and 91% for WES and amplicon based NGS.Samples with DNA yield > 100 ng had significantly higher A260/280(1.89 ± 0.32 vs 1.34 ± 0.42,P = 0.013).Tumor characteristics were not corelated with the DNA parameters.CONCLUSION EUS-FNA PDAC samples yield DNA adequate for subsequent NGS.DNA amount was similar between 22G and 19G FNA needles.DNA purity parameters may vary indirectly with needle size.
文摘Silicon phthalocyanine derivatives 1a and 1b were synthesized and characterized by UV, <sup>1</sup>H-NMR and MS. The photophysical properties of the compounds in DMSO were investigated. The maximum absorption peaks of compounds 1a and 1b at the Q-band are 681 nm. With ZnPc (Φ<sub>F</sub> = 0.20, Φ<sub>Δ</sub> = 0.67) as a reference, the fluorescence quantum yield (Φ<sub>F</sub>) of 1a and 1b are 0.20 and 0.31 respectively, and the singlet oxygen quantum yield (Φ<sub>Δ</sub>) are 0.66 and 0.59 respectively. The DNA-photocleavage activities of compounds 1a and 1b were studied by gel electrophoresis. Compounds 1a and 1b possess good photocleavage activity to pBR322 DNA. The results demonstrate that compounds 1a and 1b are potential photosensitizers for tumor therapy.
基金Thiswork was supported by the Science Creative Project of the Chinese Academy of Sciences (Grant No. KJCX2-H2).
文摘Telomeres are protein-DNA complexes at the terminals of linear chromosomes, which protect chromoso-mal integrity and maintain cellular replicative capacity. From single-cell organisms to advanced animals and plants, structures and functions of telomeres are both very conser-vative. In cells of human and vertebral animals, telomeric DNA base sequences all are (TTAGGG)n. In the present work, we have obtained absorption and fluorescence spectra measured from seven synthesized oligonucleotides to simu-late the telomeric DNA system and calculated their relative fluorescence quantum yields on which not only telomeric DNA characteristics are predicted but also possibly the shortened telomeric sequences during cell division are im-plied. Oligonucleotide 5’- TTAGGGTTAGGG holds a low relative fluorescence quantum yield and remarkable excitation energy innerconversion, which tallies with the telomeric sequence of (TTAGGG)n. This result shows that telomeric DNA has a strong non-radiative or innerconvertible capabil-