AIM: To compare the ligase detection reaction (LDR) and real-time PCR for detection of low abundant YMDD mutants in patients with chronic hepatitis B infection.METHODS: Mixtures of plasmids and serum samples from 52 c...AIM: To compare the ligase detection reaction (LDR) and real-time PCR for detection of low abundant YMDD mutants in patients with chronic hepatitis B infection.METHODS: Mixtures of plasmids and serum samples from 52 chronic hepatitis B patients with low abundant lamivudine-resistant mutations were tested with LDR and real-time PCR. Time required and reagent cost for both assays were evaluated.RESULTS: Real-time PCR detected 100, 50, 10, 1 and 0.1% of YIDD plasmid, whereas LDR detected 100, 50, 10, 1, 0.1, and 0.01% of YIDD plasmid, in mixtures with YMDD plasmid of 106 copies/mL. Among the 52 clinical serum samples, completely concordant results were obtained for all samples by both assays, and 39 YIDD, 9 YVDD, and 4 YIDD/YVDD were detected. Cost and time required for LDR and real-time PCR are 60/80 CNY (8/10.7 US dollars) and 4.5/2.5 h, respectively.CONCLUSION: LDR and real-time PCR are both sensitive and inexpensive methods for monitoring low abundant YMDD mutants during lamivudine therapy in patients with chronic hepatitis B. LDR is more sensitive and less expensive, while real-time PCR is more rapid.展开更多
AIM:To study the effects of live and dead Lactobacillus rhamnosus GG(GG) on rotavirus infection in a neonatal rat model.METHODS:At the age of 2 d,suckling Lewis rat pups were supplemented with either live or dead GG a...AIM:To study the effects of live and dead Lactobacillus rhamnosus GG(GG) on rotavirus infection in a neonatal rat model.METHODS:At the age of 2 d,suckling Lewis rat pups were supplemented with either live or dead GG and the treatment was continued daily throughout the experi-ment.At the age of 5 and 6 d the pups received oral rotavirus(RV) SA-11 strain.The pups were sacrificed at the age of 7 or 8 d by decapitation.The gastrointestinal tract was removed and macroscopic observations were done.The consistency of feces in the colon was classified using a four-tier system.RV was detected from the plasma,small intestine,colon and feces by real-time quantitative polymerase chain reaction(PCR).RESULTS:In this neonatal rat model,RV induced a mild-to-moderate diarrhea in all except one pup of the RV-inoculated rats.RV moderately reduced body weight development from day 6 onwards.On day 7,after 2 d of RV infection,live and dead GG groups gained significantly more weight than the RV group without probiotics [36%(P = 0.001) and 28%(P = 0.031),respectively].In addition,when compared with the RV control group,both live and dead GG reduced the weight ratio of colon/animal body weight to the same level as in the healthy control group,with reductions of 22%(P = 0.002) and 28%(P < 0.001),respectively.Diarrhea increased moderately in both GG groups.However,the diarrhea incidence and severity in the GG groups were not statistically significantly different as compared with the RV control group.Moreover,observed diarrhea did not provoke weight loss or death.The RV control group had the largest amount of RV PCR-positive samples among the RV-infected groups,and the live GG group had the smallest amount.Rats receiving live GG had significantly less RV in the colon(P = 0.027) when compared with the RV control group.Live GG was also more effective over dead GG in reducing the quantity of RV from plasma(P = 0.047).CONCLUSION:Both live and dead GG have beneficial effects in RV infection.GG may increase RV clearance from the body and reduce colon swelling.展开更多
Nine diploid cultivars of Italian ryegrass （Lolium multiflorum Lain.） from France （Fortyl, Vertyl and Jericho）, Germany （Ligrande）, United Kingdom （Aber Epic and Aber Mario）, Denmark （Cordelia）, Netherlands ...Nine diploid cultivars of Italian ryegrass （Lolium multiflorum Lain.） from France （Fortyl, Vertyl and Jericho）, Germany （Ligrande）, United Kingdom （Aber Epic and Aber Mario）, Denmark （Cordelia）, Netherlands （Alamo） and Poland （Tur） were tested with horizontal gel electrophoresis according to one locus （with four alleles） of the PGI enzyme system. One of them, named P4 is typical for the species, therefore can serve as a good marker for hybrids identification. Each cultivar was characterized by frequencies of different phenotypes. They were highly polymorphic （Pg = 0.58 - 0.78） and showed differences in heterozygosity level. The variability within populations （GST = 0.055） was higher than among populations （DST = 0.032）.展开更多
Objective This study aims to investigate the etiological relationship among hepatitis B virus （HBV）, hepatitis C virus （HCV）, and alcohol as risk factors in a cohort of hepatocellular carcinoma （HCC） patients fr...Objective This study aims to investigate the etiological relationship among hepatitis B virus （HBV）, hepatitis C virus （HCV）, and alcohol as risk factors in a cohort of hepatocellular carcinoma （HCC） patients from India. The clinical and biochemical profiles and tumor characteristics in the HCC cases were also evaluated. Methods A total of 357 consecutive cases of HCC fulfilling the diagnostic criteria from the Barcelona-2000 EASL conference were included in the study. The blood samples were evaluated for serological evidence of HBV and HCV infection, viral load, and genotypes using serological tests, reverse transcription-polymerase chain reaction, and restriction fragment length polymorphism. Results The male/female ratio for the HCC cases was 5.87：1. Majority of the HCC patients （33.9%） were 50 to 59 years of age, with a mean age of 4±13.23 years. More than half the cases （60.8%） had underlying cirrhosis at presentation. Among the HCC patients, 68.9% were HBV related, 21.3% were HCV related, 18.8%, were alcoholic, and 18.2% were of cryptogenic origin. The presence of any marker positive for HBV increased the risk for developing HCC by almost 27 times [OR： 27.33; （12.87-60.0）]. An increased risk of 10.6 times was observed for HCC development for cases positive for ally HCV marker [OR： 10.55; （3.13-42.73）]. Heavy alcohol consumption along with HCV RNA positivity in cirrhotic patients was found to be a risk for developing HCC by 3 folds ]OR： 3.17; （0.37-70.71）]. Conclusions Patients of chronic HBV infection followed by chronic HCV infection were at higher risk of developing HCC in India. Chronic alcohol consumption was found to be a risk factor in cirrhotic cases only when it was associated with HCV RNA positivity. Most of the patients had a large tumor size （〉5 cm） with multiple liver nodules, indicating an advanced stage of the disease thus making curative therapies difficult.展开更多
AIM:To examine the association between -86 bp(T>A) in the glucose-regulated protein 78 gene(GRP78) and hepatitis B virus(HBV) invasion.METHODS:DNA was genotyped for the single-nucleotide polymorphism by polymerase ...AIM:To examine the association between -86 bp(T>A) in the glucose-regulated protein 78 gene(GRP78) and hepatitis B virus(HBV) invasion.METHODS:DNA was genotyped for the single-nucleotide polymorphism by polymerase chain reaction followed by sequencing in a sample of 382 unrelated HBV carriers and a total of 350 sex-and age-matched healthy controls.Serological markers for HBV infection were determined by enzyme-linked immunosorbent as-say kits or clinical chemistry testing.RESULTS:The distributions of allelotype and genotype in cases were not significantly different from those in controls.In addition,our fi ndings suggested that neither alanine aminotransferase/hepatitis B e antigen nor HBV-DNA were associated with the allele/genotype variation in HBV infected individuals.CONCLUSION:-86 bp T>A polymorphism in GRP78 gene is not related to the clinical risk and acute exacerbation of HBV invasion.展开更多
文摘AIM: To compare the ligase detection reaction (LDR) and real-time PCR for detection of low abundant YMDD mutants in patients with chronic hepatitis B infection.METHODS: Mixtures of plasmids and serum samples from 52 chronic hepatitis B patients with low abundant lamivudine-resistant mutations were tested with LDR and real-time PCR. Time required and reagent cost for both assays were evaluated.RESULTS: Real-time PCR detected 100, 50, 10, 1 and 0.1% of YIDD plasmid, whereas LDR detected 100, 50, 10, 1, 0.1, and 0.01% of YIDD plasmid, in mixtures with YMDD plasmid of 106 copies/mL. Among the 52 clinical serum samples, completely concordant results were obtained for all samples by both assays, and 39 YIDD, 9 YVDD, and 4 YIDD/YVDD were detected. Cost and time required for LDR and real-time PCR are 60/80 CNY (8/10.7 US dollars) and 4.5/2.5 h, respectively.CONCLUSION: LDR and real-time PCR are both sensitive and inexpensive methods for monitoring low abundant YMDD mutants during lamivudine therapy in patients with chronic hepatitis B. LDR is more sensitive and less expensive, while real-time PCR is more rapid.
基金Supported by Valio Ltd and the Finnish Funding Agency of Technology and InnovationThe Foundation for Nutrition Research has financially contributed to Liisa Lehtoranta’s doctoral studies
文摘AIM:To study the effects of live and dead Lactobacillus rhamnosus GG(GG) on rotavirus infection in a neonatal rat model.METHODS:At the age of 2 d,suckling Lewis rat pups were supplemented with either live or dead GG and the treatment was continued daily throughout the experi-ment.At the age of 5 and 6 d the pups received oral rotavirus(RV) SA-11 strain.The pups were sacrificed at the age of 7 or 8 d by decapitation.The gastrointestinal tract was removed and macroscopic observations were done.The consistency of feces in the colon was classified using a four-tier system.RV was detected from the plasma,small intestine,colon and feces by real-time quantitative polymerase chain reaction(PCR).RESULTS:In this neonatal rat model,RV induced a mild-to-moderate diarrhea in all except one pup of the RV-inoculated rats.RV moderately reduced body weight development from day 6 onwards.On day 7,after 2 d of RV infection,live and dead GG groups gained significantly more weight than the RV group without probiotics [36%(P = 0.001) and 28%(P = 0.031),respectively].In addition,when compared with the RV control group,both live and dead GG reduced the weight ratio of colon/animal body weight to the same level as in the healthy control group,with reductions of 22%(P = 0.002) and 28%(P < 0.001),respectively.Diarrhea increased moderately in both GG groups.However,the diarrhea incidence and severity in the GG groups were not statistically significantly different as compared with the RV control group.Moreover,observed diarrhea did not provoke weight loss or death.The RV control group had the largest amount of RV PCR-positive samples among the RV-infected groups,and the live GG group had the smallest amount.Rats receiving live GG had significantly less RV in the colon(P = 0.027) when compared with the RV control group.Live GG was also more effective over dead GG in reducing the quantity of RV from plasma(P = 0.047).CONCLUSION:Both live and dead GG have beneficial effects in RV infection.GG may increase RV clearance from the body and reduce colon swelling.
文摘Nine diploid cultivars of Italian ryegrass （Lolium multiflorum Lain.） from France （Fortyl, Vertyl and Jericho）, Germany （Ligrande）, United Kingdom （Aber Epic and Aber Mario）, Denmark （Cordelia）, Netherlands （Alamo） and Poland （Tur） were tested with horizontal gel electrophoresis according to one locus （with four alleles） of the PGI enzyme system. One of them, named P4 is typical for the species, therefore can serve as a good marker for hybrids identification. Each cultivar was characterized by frequencies of different phenotypes. They were highly polymorphic （Pg = 0.58 - 0.78） and showed differences in heterozygosity level. The variability within populations （GST = 0.055） was higher than among populations （DST = 0.032）.
文摘Objective This study aims to investigate the etiological relationship among hepatitis B virus （HBV）, hepatitis C virus （HCV）, and alcohol as risk factors in a cohort of hepatocellular carcinoma （HCC） patients from India. The clinical and biochemical profiles and tumor characteristics in the HCC cases were also evaluated. Methods A total of 357 consecutive cases of HCC fulfilling the diagnostic criteria from the Barcelona-2000 EASL conference were included in the study. The blood samples were evaluated for serological evidence of HBV and HCV infection, viral load, and genotypes using serological tests, reverse transcription-polymerase chain reaction, and restriction fragment length polymorphism. Results The male/female ratio for the HCC cases was 5.87：1. Majority of the HCC patients （33.9%） were 50 to 59 years of age, with a mean age of 4±13.23 years. More than half the cases （60.8%） had underlying cirrhosis at presentation. Among the HCC patients, 68.9% were HBV related, 21.3% were HCV related, 18.8%, were alcoholic, and 18.2% were of cryptogenic origin. The presence of any marker positive for HBV increased the risk for developing HCC by almost 27 times [OR： 27.33; （12.87-60.0）]. An increased risk of 10.6 times was observed for HCC development for cases positive for ally HCV marker [OR： 10.55; （3.13-42.73）]. Heavy alcohol consumption along with HCV RNA positivity in cirrhotic patients was found to be a risk for developing HCC by 3 folds ]OR： 3.17; （0.37-70.71）]. Conclusions Patients of chronic HBV infection followed by chronic HCV infection were at higher risk of developing HCC in India. Chronic alcohol consumption was found to be a risk factor in cirrhotic cases only when it was associated with HCV RNA positivity. Most of the patients had a large tumor size （〉5 cm） with multiple liver nodules, indicating an advanced stage of the disease thus making curative therapies difficult.
基金Supported by The grant from Ministry of Science and Technology of China, No. 2006CB910104the Foundation of Guangzhou Science and Technology Bureauthe People’s Republic of China, No. 2005Z1-E0131
文摘AIM:To examine the association between -86 bp(T>A) in the glucose-regulated protein 78 gene(GRP78) and hepatitis B virus(HBV) invasion.METHODS:DNA was genotyped for the single-nucleotide polymorphism by polymerase chain reaction followed by sequencing in a sample of 382 unrelated HBV carriers and a total of 350 sex-and age-matched healthy controls.Serological markers for HBV infection were determined by enzyme-linked immunosorbent as-say kits or clinical chemistry testing.RESULTS:The distributions of allelotype and genotype in cases were not significantly different from those in controls.In addition,our fi ndings suggested that neither alanine aminotransferase/hepatitis B e antigen nor HBV-DNA were associated with the allele/genotype variation in HBV infected individuals.CONCLUSION:-86 bp T>A polymorphism in GRP78 gene is not related to the clinical risk and acute exacerbation of HBV invasion.
