AIM: To investigate the pattern of mucin expression and concentration in bile obtained during endoscopic retrograde cholangiography (ERC) in relation to gallstone disease.METHODS: Bile samples obtained at ERC from...AIM: To investigate the pattern of mucin expression and concentration in bile obtained during endoscopic retrograde cholangiography (ERC) in relation to gallstone disease.METHODS: Bile samples obtained at ERC from 29 consecutive patients, 17 with and 12 without gallstone disease were evaluated for mucin content by gel filtration on a Sepharose CL-4B column. Dot blot analysis for bile mucin apoproteins was performed with antibodies to Mucin 1 (MUC1), MUC2, MUC3, MUC5AC, MUC5B and MUC6. Staining intensity score (0-3) was used as a measure of antigen expression.RESULTS: MUCl, MUC2, MUC3, MUCSAC, MUC5B and MUC6 were demonstrated in 34.4%, 34.4%, 51.7%, 51.7%, 55.1% and 27.5% of bile samples, respectively.The staining intensity scores were 0.62 ± 0.94, 0.58 ± 0.90, 0.79 ± 0.97, 1.06 ± 1.22, 1.20 ± 2.26 and 0.41 ± 0.73, respectively. Mean mucin concentration measured in bile by the Sepharose CL-4B method was 22.8 ± 24.0 mg/mL (range 3.4-89.0 mg/mL). Mean protein concentration was 8.1 ± 4.8 mg/mL (range 1.7-23.2 mg/mL).CONCLUSION: High levels of MUC3, MUC5AC and MUC5B are expressed in bile aspirated during ERC examination. A specific pattern of mucin gene expression or change in mucin concentration was not found in gallstone disease.展开更多
AIM: TO determine whether -1195 A→G and/or -765 G→C polymorphisms in Cyclooxygenase-2 CCOX-2) may have a risk modifying effect on the development of esophageal carcinoma in a Dutch Caucasian population. METHODS: ...AIM: TO determine whether -1195 A→G and/or -765 G→C polymorphisms in Cyclooxygenase-2 CCOX-2) may have a risk modifying effect on the development of esophageal carcinoma in a Dutch Caucasian population. METHODS: Two study groups were recruited, 252 patients with esophageal carcinoma and 240 healthy controls, matched for race, age, gender and recruiting area. DNA was isolated from whole blood and used for genotyping. PCR products were digested with restriction enzymes and products were analyzed by agarose gel electrophoresis. Odds ratios (OR) and 95% confidence intervals (CI) were estimated. RESULTS: The distribution of the -1195A→G polymorphism was significantly different in esophageal cancer patients compared to controls. The -1195 GG genotype resulted in a higher risk of developing esophageal adenocarcinoma (OR = 3.85, 95% CI: 1.45-10.3) compared with the -1195AA genotype as a reference. The -765 G→C genotype distribution was not different between the two groups. The GG/ GG haplotype was present more often in esophageal adenocarcinoma patients than in controls (OR = 3.45, 95% CI: 1.24-9.58; with AG/AG as a reference). The same trends were observed in patients with squamous cell carcinomas, however, the results did not reach statistical significance. CONCLUSION: Presence of the COX-2 -1195 GG genotype and of the GG/GG haplotype may result in a higher risk of developing esophageal carcinoma.展开更多
The purpose of this study was to develop a biological model to evaluate the genotoxic effects of natural emissions of Radon-222 and its decay products. To this aim, mice of the Swiss CD1 strain were exposed to Radon f...The purpose of this study was to develop a biological model to evaluate the genotoxic effects of natural emissions of Radon-222 and its decay products. To this aim, mice of the Swiss CD1 strain were exposed to Radon for different periods (adult life, early postnatal and in utero exposure) and two simple but accurate mutagenicity tests (Micronucleus test and the Comet assay) were applied to the peripheral blood of mice. The study was carried out in two small towns in Latium region--ltaly, where radon pollution is notoriously present due to the volcanic soils. One experiment was carried out in the cellar of a house in Ciampino (Rome) and two experiments were performed in an old cellar in Vetralla (Viterbo). The results showed that in all mice groups exposed to natural emissions of radon and its decay products, the micronucleated erythrocytes frequency (ME) was significantly higher than that observed in the mice control. The single cell gel electrophoresis (Comet assay) was performed in lymphocytes of adult mice in the last experiment. The results for this test also show that DNA damage was higher than in the cells of the mice control and the cells of mice exposed for a shorter period of time. To confirm these findings, the single cell gel electrophoresis (Comet assay) was performed in lymphocytes of adult mice in the last experiment. Similarly, this result could be linked to a greater sensitivity of neonatal mice to radon emissions compared with intrauterine mice. Further investigations on the effects of radon and its decay products during the intrauterine life and the first neonatal period should be performed to better clarify its genotoxic activity.展开更多
A microfluidic approach to generate hydrogel microstructures inside microchannels for controlled encapsulation of single cells was developed. The method was based,on a modified microscope projection photolithography w...A microfluidic approach to generate hydrogel microstructures inside microchannels for controlled encapsulation of single cells was developed. The method was based,on a modified microscope projection photolithography which allowed for the photopolymerization of poly(ethylene glycol) diacrylate (PEGDA) inside microchannels. Uniformsized hydrogel microstmctures (-50 pan in diameter) were generated one by one with determined positions to encapsulate single cells without losing the viability. Cells of interest could be identified by any kinds of visible labels to be selectively encapsulated inside the formed hydrogel microstructures. Large-scale encapsulation of single cells was achieved with a relatively high efficiency of 80% and the viability of encapsulated cells could be guaranteed by removing the dead cells identified with Trypan blue. This method is simple, fast and convenient to pattern the microchannels with single cells for a wide range of cell-based applications. For demonstration, two intracellular enzyme assays of carboxylesterase were performed to investigate the distribution of enzyme concentrations and the kinetic information within the encapsulated single HepG2 cells.展开更多
文摘AIM: To investigate the pattern of mucin expression and concentration in bile obtained during endoscopic retrograde cholangiography (ERC) in relation to gallstone disease.METHODS: Bile samples obtained at ERC from 29 consecutive patients, 17 with and 12 without gallstone disease were evaluated for mucin content by gel filtration on a Sepharose CL-4B column. Dot blot analysis for bile mucin apoproteins was performed with antibodies to Mucin 1 (MUC1), MUC2, MUC3, MUC5AC, MUC5B and MUC6. Staining intensity score (0-3) was used as a measure of antigen expression.RESULTS: MUCl, MUC2, MUC3, MUCSAC, MUC5B and MUC6 were demonstrated in 34.4%, 34.4%, 51.7%, 51.7%, 55.1% and 27.5% of bile samples, respectively.The staining intensity scores were 0.62 ± 0.94, 0.58 ± 0.90, 0.79 ± 0.97, 1.06 ± 1.22, 1.20 ± 2.26 and 0.41 ± 0.73, respectively. Mean mucin concentration measured in bile by the Sepharose CL-4B method was 22.8 ± 24.0 mg/mL (range 3.4-89.0 mg/mL). Mean protein concentration was 8.1 ± 4.8 mg/mL (range 1.7-23.2 mg/mL).CONCLUSION: High levels of MUC3, MUC5AC and MUC5B are expressed in bile aspirated during ERC examination. A specific pattern of mucin gene expression or change in mucin concentration was not found in gallstone disease.
文摘AIM: TO determine whether -1195 A→G and/or -765 G→C polymorphisms in Cyclooxygenase-2 CCOX-2) may have a risk modifying effect on the development of esophageal carcinoma in a Dutch Caucasian population. METHODS: Two study groups were recruited, 252 patients with esophageal carcinoma and 240 healthy controls, matched for race, age, gender and recruiting area. DNA was isolated from whole blood and used for genotyping. PCR products were digested with restriction enzymes and products were analyzed by agarose gel electrophoresis. Odds ratios (OR) and 95% confidence intervals (CI) were estimated. RESULTS: The distribution of the -1195A→G polymorphism was significantly different in esophageal cancer patients compared to controls. The -1195 GG genotype resulted in a higher risk of developing esophageal adenocarcinoma (OR = 3.85, 95% CI: 1.45-10.3) compared with the -1195AA genotype as a reference. The -765 G→C genotype distribution was not different between the two groups. The GG/ GG haplotype was present more often in esophageal adenocarcinoma patients than in controls (OR = 3.45, 95% CI: 1.24-9.58; with AG/AG as a reference). The same trends were observed in patients with squamous cell carcinomas, however, the results did not reach statistical significance. CONCLUSION: Presence of the COX-2 -1195 GG genotype and of the GG/GG haplotype may result in a higher risk of developing esophageal carcinoma.
文摘The purpose of this study was to develop a biological model to evaluate the genotoxic effects of natural emissions of Radon-222 and its decay products. To this aim, mice of the Swiss CD1 strain were exposed to Radon for different periods (adult life, early postnatal and in utero exposure) and two simple but accurate mutagenicity tests (Micronucleus test and the Comet assay) were applied to the peripheral blood of mice. The study was carried out in two small towns in Latium region--ltaly, where radon pollution is notoriously present due to the volcanic soils. One experiment was carried out in the cellar of a house in Ciampino (Rome) and two experiments were performed in an old cellar in Vetralla (Viterbo). The results showed that in all mice groups exposed to natural emissions of radon and its decay products, the micronucleated erythrocytes frequency (ME) was significantly higher than that observed in the mice control. The single cell gel electrophoresis (Comet assay) was performed in lymphocytes of adult mice in the last experiment. The results for this test also show that DNA damage was higher than in the cells of the mice control and the cells of mice exposed for a shorter period of time. To confirm these findings, the single cell gel electrophoresis (Comet assay) was performed in lymphocytes of adult mice in the last experiment. Similarly, this result could be linked to a greater sensitivity of neonatal mice to radon emissions compared with intrauterine mice. Further investigations on the effects of radon and its decay products during the intrauterine life and the first neonatal period should be performed to better clarify its genotoxic activity.
基金supported by the National Natural Science Foundation of China (20935002 & 90813015)
文摘A microfluidic approach to generate hydrogel microstructures inside microchannels for controlled encapsulation of single cells was developed. The method was based,on a modified microscope projection photolithography which allowed for the photopolymerization of poly(ethylene glycol) diacrylate (PEGDA) inside microchannels. Uniformsized hydrogel microstmctures (-50 pan in diameter) were generated one by one with determined positions to encapsulate single cells without losing the viability. Cells of interest could be identified by any kinds of visible labels to be selectively encapsulated inside the formed hydrogel microstructures. Large-scale encapsulation of single cells was achieved with a relatively high efficiency of 80% and the viability of encapsulated cells could be guaranteed by removing the dead cells identified with Trypan blue. This method is simple, fast and convenient to pattern the microchannels with single cells for a wide range of cell-based applications. For demonstration, two intracellular enzyme assays of carboxylesterase were performed to investigate the distribution of enzyme concentrations and the kinetic information within the encapsulated single HepG2 cells.