Objectives: To obtain very end full-length cDNA ofhepatitis C virus (HCV) 5’ untranslated region(5’UTR) and analyze its primary and secondarystructure.Methods: A patient infected genotype 2a HCV wasidentified by rev...Objectives: To obtain very end full-length cDNA ofhepatitis C virus (HCV) 5’ untranslated region(5’UTR) and analyze its primary and secondarystructure.Methods: A patient infected genotype 2a HCV wasidentified by reverse transcription-nested polymerasechain reaction (RT-PCR) and restriction fragmentlength polymorphism (RFLP). Total RNA isolatedfrom the serum was used as template, and the cDNAof the 5’ untranslated region was amplified using rap-id amplification of cDNA ends (RACE). The frag-ments were recombinated by A-T clone strategy, andthe recombinants were confirmed by RFLP andPCR, and sequenced subsequently. Secondary struc-tures were analysed by RNAdraw.Results: Very end full-length cDNA of genotype 2aHCV 5’ UTR was obtained by RACE. In five clonesobtained, three contained full-length 5’UTR cDNA;A21G, G170A, T222C, T247C, C339T substitutionswere found as compared to HC-J6. Homological re-sults of HCV-1, HC-J6, HC-C2, HC-J8 were 93.6%-94.4%, 92.1%-93%, 98.8%-99.7%, 96.2%-96.5%, respectively; however, the substitutions didnot alter secondary structure. Two of 5 clones weredeletions of 53bp and 135bp at the 5’terminal ofHCV 5’UTR, respectively.Conclusions: RACE can be used to obtain the full-length cDNA of 2a genotype HCV 5’UTR. Genes de-leted at the 5’ terminal of HCV circulate in hepatitisC patients.展开更多
There are numerous studies on polar amplification and its influence on mid-latitude weather and climate.However,assessments on whether polar amplification occurs in Antarctica are rarely conducted.Based on the latest ...There are numerous studies on polar amplification and its influence on mid-latitude weather and climate.However,assessments on whether polar amplification occurs in Antarctica are rarely conducted.Based on the latest atmospheric reanalysis of ERA5 produced by European Centre for Medium-Range Weather Forecasts(ECMWF),we have defined the Antarctic amplification index,and calculated the trend of annual and seasonal Surface Air Temperature(SAT)mean during 1979-2019 for Antarctic Ice Sheet(AIS)and the trend mean of different meridional sectors of Antarctic sub regions including East Antarctic Ice Sheet(EAIS),West Antarctic Ice Sheet(WAIS)and Antarctic Peninsula(AP).Antarctic amplification shows regional differences and seasonal variations.Antarctica shows a slight warming with the largest magnitude in AP.The temperature anomalies indicate the least fluctuations in austral summer,and the more fluctuations in winter and spring.In austral summer,the warming trend domains EAIS and WAIS,while the cooling trend appears over AP.The zonal mean in Southern Hemisphere maintains a warming trend in the low latitudes,and fluctuates greatly in the middle and high latitudes.The strongest Antarctic amplification phenomenon occurs in spring,with the amplification index of 1.20.For AP,the amplification occurs in austral autumn,and the amplification index is 2.16.At South Pole and the surrounding regions,SAT for land only fluctuates largely and shows different trends in different seasons.The mechanism of Antarctic amplification is unclear till now,and its research suffers from the limitation of measured data.This suggests that future research needs progress in comprehensive ground observation network,remote sensing data accumulation,and high-resolution climate modeling with better representation of both atmospheric and cryospheric processes in Antarctica.展开更多
AIM: To investigate the frequency of mutations in pre-core (pre-C) and basic core promoter (BCP) regions of hepatitis B virus (HBV) from Shanxi Province, and the association between mutations and disease related index...AIM: To investigate the frequency of mutations in pre-core (pre-C) and basic core promoter (BCP) regions of hepatitis B virus (HBV) from Shanxi Province, and the association between mutations and disease related indexes.METHODS: One hundred chronic hepatitis B patients treated at Shanxi Province Hospital of Traditional Chinese Medicine were included in this study. PCR-reverse dot blot hybridization and mismatch amplification mutation assay (MAMA)-PCR were used to detect the mutations in the HBV pre-C and BCP regions. HBV DNA content and liver function were compared between patients with mutant HBV pre-C and BCP loci and those with wild-type loci. The consistency between PCR-reverse dot blot hybridization and MAMA-PCR for detecting mutations in the HBV pre-C and BCP regions was assessed.RESULTS: Of the 100 serum samples detected, 9.38% had single mutations in the pre-C region, 29.17% had single mutations in the BCP region, 41.67% had mutations in both BCP and pre-C regions, and 19.79% had wild-type loci. The rates of BCP and pre-C mutations were 65.7% and 34.3%, respectively, in hepatitis B e antigen (HBeAg) positive patients, and 84.6% and 96.2%, respectively, in HBeAg negative patients. The rate of pre-C mutations was significantly higher in HBeAg negative patients than in HBeAg positive patients (χ<sup>2</sup> = 26.62, P = 0.00), but there was no significant difference in the distribution of mutations in the BCP region between HBeAg positive and negative patients (χ<sup>2</sup> = 2.43, P = 0.12). The presence of mutations in the pre-C (Wilcoxon W = 1802.5, P = 0.00) and BCP regions (Wilcoxon W = 2906.5, P = 0.00) was more common in patients with low HBV DNA content. Both AST and GGT were significantly higher in patients with mutant pre-C and BCP loci than in those with wild-type loci (P < 0.05). PCR-reverse dot blot hybridization and MAMA-PCR for detection of mutations in the BCP and pre-C regions had good consistency, and the Kappa values obtained were 0.91 and 0.58, respectively.CONCLUSION: HBeAg negative patients tend to have HBV pre-C mutations. However, these mutations do not cause increased DNA copies, but associate with damage of liver function.展开更多
Long-term probabilistic prediction of extreme rainfall at the regional scale is a significant tool in the mitigation of hydro-geological disasters: it actually provides the starting point in the design of strategic hy...Long-term probabilistic prediction of extreme rainfall at the regional scale is a significant tool in the mitigation of hydro-geological disasters: it actually provides the starting point in the design of strategic hydraulic infrastructures and emergency plans. A crucial task of regional estimation of extreme rainfall is how to include the complex effects of orographic barriers in a mathematical model for Intensity-Duration-Frequency (IDF) curves. Here, an analysis of how orography can affect extreme rainfall at different durations is presented for three orographic systems that are very relevant for hydrological risk assessment in the Campania Region in Southern Italy. Then, we introduce a power law model to link the amplification factor to the duration, thus allowing a simple and effective enhancement of the IDF model in mountainous areas.展开更多
为了优化以土壤微生物DNA为模板的PCR反应条件,采用E.Z.N.A.soil DNA kit试剂盒提取土壤微生物总DNA,对16SrDNA V3可变区的PCR反应体系和反应条件进行优化。主要从DNA模板用量、引物浓度、退火温度、热启动方式4个方面进行筛选试验,最...为了优化以土壤微生物DNA为模板的PCR反应条件,采用E.Z.N.A.soil DNA kit试剂盒提取土壤微生物总DNA,对16SrDNA V3可变区的PCR反应体系和反应条件进行优化。主要从DNA模板用量、引物浓度、退火温度、热启动方式4个方面进行筛选试验,最后得出最适宜的土壤DNA扩增体系为:10.5ng模板DNA、5μL 10×buffer、4μL 2.5mmol/L dNTP、10μmol/L引物各1.5μL,2.5UTaq酶,加无菌ddH2O补足至50μL;PCR循环程序为:94℃预变性5min;94℃变性1min,52℃退火1min,72℃延伸70s,29个循环;72℃延伸5min。试验结果表明:选择热启动方式和合适的退火温度是获得高质量PCR产物的关键。展开更多
基金This work was supported by two grants from National Science Foundation of China (No: 39770684, 30170844).
文摘Objectives: To obtain very end full-length cDNA ofhepatitis C virus (HCV) 5’ untranslated region(5’UTR) and analyze its primary and secondarystructure.Methods: A patient infected genotype 2a HCV wasidentified by reverse transcription-nested polymerasechain reaction (RT-PCR) and restriction fragmentlength polymorphism (RFLP). Total RNA isolatedfrom the serum was used as template, and the cDNAof the 5’ untranslated region was amplified using rap-id amplification of cDNA ends (RACE). The frag-ments were recombinated by A-T clone strategy, andthe recombinants were confirmed by RFLP andPCR, and sequenced subsequently. Secondary struc-tures were analysed by RNAdraw.Results: Very end full-length cDNA of genotype 2aHCV 5’ UTR was obtained by RACE. In five clonesobtained, three contained full-length 5’UTR cDNA;A21G, G170A, T222C, T247C, C339T substitutionswere found as compared to HC-J6. Homological re-sults of HCV-1, HC-J6, HC-C2, HC-J8 were 93.6%-94.4%, 92.1%-93%, 98.8%-99.7%, 96.2%-96.5%, respectively; however, the substitutions didnot alter secondary structure. Two of 5 clones weredeletions of 53bp and 135bp at the 5’terminal ofHCV 5’UTR, respectively.Conclusions: RACE can be used to obtain the full-length cDNA of 2a genotype HCV 5’UTR. Genes de-leted at the 5’ terminal of HCV circulate in hepatitisC patients.
基金supported by the National Natural Science Foundation of China(Grant No.41671073,41476164)the Freedom Project of the State Key Laboratory of Cryosphere Science,Northwest Institute of Eco-Environment and Resources,CAS(Grant SKLCS-ZZ-2021)。
文摘There are numerous studies on polar amplification and its influence on mid-latitude weather and climate.However,assessments on whether polar amplification occurs in Antarctica are rarely conducted.Based on the latest atmospheric reanalysis of ERA5 produced by European Centre for Medium-Range Weather Forecasts(ECMWF),we have defined the Antarctic amplification index,and calculated the trend of annual and seasonal Surface Air Temperature(SAT)mean during 1979-2019 for Antarctic Ice Sheet(AIS)and the trend mean of different meridional sectors of Antarctic sub regions including East Antarctic Ice Sheet(EAIS),West Antarctic Ice Sheet(WAIS)and Antarctic Peninsula(AP).Antarctic amplification shows regional differences and seasonal variations.Antarctica shows a slight warming with the largest magnitude in AP.The temperature anomalies indicate the least fluctuations in austral summer,and the more fluctuations in winter and spring.In austral summer,the warming trend domains EAIS and WAIS,while the cooling trend appears over AP.The zonal mean in Southern Hemisphere maintains a warming trend in the low latitudes,and fluctuates greatly in the middle and high latitudes.The strongest Antarctic amplification phenomenon occurs in spring,with the amplification index of 1.20.For AP,the amplification occurs in austral autumn,and the amplification index is 2.16.At South Pole and the surrounding regions,SAT for land only fluctuates largely and shows different trends in different seasons.The mechanism of Antarctic amplification is unclear till now,and its research suffers from the limitation of measured data.This suggests that future research needs progress in comprehensive ground observation network,remote sensing data accumulation,and high-resolution climate modeling with better representation of both atmospheric and cryospheric processes in Antarctica.
基金Supported by Youth Foundation of Health and Family Planning Commission of Shanxi ProvinceNo.201301024
文摘AIM: To investigate the frequency of mutations in pre-core (pre-C) and basic core promoter (BCP) regions of hepatitis B virus (HBV) from Shanxi Province, and the association between mutations and disease related indexes.METHODS: One hundred chronic hepatitis B patients treated at Shanxi Province Hospital of Traditional Chinese Medicine were included in this study. PCR-reverse dot blot hybridization and mismatch amplification mutation assay (MAMA)-PCR were used to detect the mutations in the HBV pre-C and BCP regions. HBV DNA content and liver function were compared between patients with mutant HBV pre-C and BCP loci and those with wild-type loci. The consistency between PCR-reverse dot blot hybridization and MAMA-PCR for detecting mutations in the HBV pre-C and BCP regions was assessed.RESULTS: Of the 100 serum samples detected, 9.38% had single mutations in the pre-C region, 29.17% had single mutations in the BCP region, 41.67% had mutations in both BCP and pre-C regions, and 19.79% had wild-type loci. The rates of BCP and pre-C mutations were 65.7% and 34.3%, respectively, in hepatitis B e antigen (HBeAg) positive patients, and 84.6% and 96.2%, respectively, in HBeAg negative patients. The rate of pre-C mutations was significantly higher in HBeAg negative patients than in HBeAg positive patients (χ<sup>2</sup> = 26.62, P = 0.00), but there was no significant difference in the distribution of mutations in the BCP region between HBeAg positive and negative patients (χ<sup>2</sup> = 2.43, P = 0.12). The presence of mutations in the pre-C (Wilcoxon W = 1802.5, P = 0.00) and BCP regions (Wilcoxon W = 2906.5, P = 0.00) was more common in patients with low HBV DNA content. Both AST and GGT were significantly higher in patients with mutant pre-C and BCP loci than in those with wild-type loci (P < 0.05). PCR-reverse dot blot hybridization and MAMA-PCR for detection of mutations in the BCP and pre-C regions had good consistency, and the Kappa values obtained were 0.91 and 0.58, respectively.CONCLUSION: HBeAg negative patients tend to have HBV pre-C mutations. However, these mutations do not cause increased DNA copies, but associate with damage of liver function.
文摘Long-term probabilistic prediction of extreme rainfall at the regional scale is a significant tool in the mitigation of hydro-geological disasters: it actually provides the starting point in the design of strategic hydraulic infrastructures and emergency plans. A crucial task of regional estimation of extreme rainfall is how to include the complex effects of orographic barriers in a mathematical model for Intensity-Duration-Frequency (IDF) curves. Here, an analysis of how orography can affect extreme rainfall at different durations is presented for three orographic systems that are very relevant for hydrological risk assessment in the Campania Region in Southern Italy. Then, we introduce a power law model to link the amplification factor to the duration, thus allowing a simple and effective enhancement of the IDF model in mountainous areas.
