The optimal antisense accessible sites （AAS） of uroplakin Ⅱ （UP Ⅱ ） mRNA, a specific gene expressed in bladder urothelium, were selected in order to provide a novel method for targeted therapy of transitional ce...The optimal antisense accessible sites （AAS） of uroplakin Ⅱ （UP Ⅱ ） mRNA, a specific gene expressed in bladder urothelium, were selected in order to provide a novel method for targeted therapy of transitional cell carcinoma （TCC） of bladder. The 20 mer random oligonucleotide library was synthesized, hybridized with in vitro transcripted total UPⅡ cRNA, then digested by RNase H. After primer extension and autoradiography, the AAS of UPⅡ were selected. The RNADraw soft- ware was used to analyze and choose the AAS with obvious stem-loop structures, according to which the complementary antisense oligonucleotides （AS-ODN） were synthesized. With the AS-ODN0 designed only by RNADraw as a control, the AS-ODNs were transferred into UP Ⅱ highly-expressing cell line RT4. The cellular expression of UPⅡ mRNA was detected by RT-PCR and Western blot. Twelve AAS of UPⅡ mRNA were selected in vitro. Four AAS with stem-loop structures were chosen, locating at 558-577, 552-571, 217-236 and 97-116 bp of UP Ⅱ mRNA respectively. After transfection with the corresponding AS-ODN （AS-ODN1, AS-ODN2, AS-ODN3 and AS-ODN4） for 18 h, the UPⅡ mRNA levels in RT4cells were reduced by 29,3%, 82.7%, 71.3% and 70.9%, while UP Ⅱ protein levels were decreased by 20.2%, 78.5%, 65.2% and 64.4% respectively, which were significantly higher than those of AS-ODN0 （14.3%, 12.1% respectively） （P〈0.01）. The AAS of UPⅡ mRNA was effectively selected in vitro by random oligonucletide library/RNase H cleavage method in combination with computer software analysis, which had important reference values for further studying biological functions of UP Ⅱ gene and targeted therapeutic strategy for TCC of bladder.展开更多
Building codes have widely considered the shear wave velocity to make a reliable subsoil seismic classification, based on the knowledge of the mechanical properties of material deposits down to bedrock. This approach ...Building codes have widely considered the shear wave velocity to make a reliable subsoil seismic classification, based on the knowledge of the mechanical properties of material deposits down to bedrock. This approach has limitations because geophysical data are often very expensive to obtain. Recently, other alternatives have been proposed based on measurements of background noise and estimation of the H/V amplification curve. However, the use of this technique needs a regulatory framework before it can become a realistic site classification procedure. This paper proposes a new formulation for characterizing design sites in accordance with the Algerian seismic building code (RPA99/ver.2003), through transfer functions, by following a stochastic approach combined to a statistical study. For each soil type, the deterministic calculation of the average transfer function is performed over a wide sample of 1-D soil profiles, where the average shear wave (S-W) velocity, V<sub>s</sub>, in soil layers is simulated using random field theory. Average transfer functions are also used to calculate average site factors and normalized acceleration response spectra to highlight the amplification potential of each site type, since frequency content of the transfer function is significantly similar to that of the H/V amplification curve. Comparison is done with the RPA99/ver.2003 and Eurocode8 (EC8) design response spectra, respectively. In the absence of geophysical data, the proposed classification approach together with micro-tremor measures can be used toward a better soil classification.展开更多
目的采用两样本孟德尔随机化(MR)探讨417种循环代谢产物与吉兰-巴雷综合征(GBS)风险的因果关系。方法通过MRC IEU OpenGWAS项目获得3个循环代谢产物全基因组关联研究(GWAS)数据,分别为“met-a”“met-c”和“met-d”。GBS相关的单核苷...目的采用两样本孟德尔随机化(MR)探讨417种循环代谢产物与吉兰-巴雷综合征(GBS)风险的因果关系。方法通过MRC IEU OpenGWAS项目获得3个循环代谢产物全基因组关联研究(GWAS)数据,分别为“met-a”“met-c”和“met-d”。GBS相关的单核苷酸多态性位点(SNPs)数据来源于芬兰生物银行数据库,表型代码为“finn-b-G6_GUILBAR”。将来自GWAS的与循环代谢产物密切相关的遗传变异数据(SNPs)作为工具变量(IVs),与来自芬兰的GBS GWAS数据进行双样本MR分析,主要采用随机效应模型的逆方差加权(IVW)方法,根据效应指标优势比(OR)和95%CI评估结果。使用留一法、异质性检验、水平基因多效性检验验证结果的稳定性和可靠性。结果共5种循环代谢产物具有与GBS因果关系的提示性证据。其中,肌酐(OR=2.924,95%CI:1.194~7.163,P=0.019)、谷氨酰胺(OR=1.902,95%CI:1.007~3.592,P=0.048)、异戊酰基肉碱(OR=140.767,95%CI:3.510~5645.336,P=0.009)的循环水平与GBS风险较高有关。相反,基因预测葡萄糖(OR=0.308,95%CI:0.010~0.981,P=0.046)、X-11491(OR=0.069,95%CI:0.007~0.707,P=0.024)的循环水平与GBS风险呈负相关。结论肌酐、谷氨酰胺、异戊酰基肉碱、葡萄糖、X-11491可能与GBS有因果关系。展开更多
α-Amylase is widely used in biotechnology industries such as food,textile and detergent.It occupies an important position in the domestic and international enzyme market.An important reason that amylase can achieve i...α-Amylase is widely used in biotechnology industries such as food,textile and detergent.It occupies an important position in the domestic and international enzyme market.An important reason that amylase can achieve its function is it has catalysis domain,which is a very basilic domain,and the function of the catalysis domain mainly depends on catalytic active site.So changing the base of the active site or the base around the active site may affect a lot on α-amylase.This article mainly describes using random mutagenesis of the α-amylase by error-prone PCR,detecting the amylase activity of the mutant strains,and then sequencing the mutants.8 available mutants was picked to do further analysis and prediction of 3D structure.Choosing the mutant with the highest enzyme activity improvement as a further research target molecule,the potential structure changes were predicted by replacement of amino acid residue at the mutant site with other 18 residues.展开更多
Background:Large area forest inventories often use regular grids(with a single random start)of sample locations to ensure a uniform sampling intensity across the space of the surveyed populations.A design-unbiased est...Background:Large area forest inventories often use regular grids(with a single random start)of sample locations to ensure a uniform sampling intensity across the space of the surveyed populations.A design-unbiased estimator of variance does not exist for this design.Oftentimes,a quasi-default estimator applicable to simple random sampling(SRS)is used,even if it carries with it the likely risk of overestimating the variance by a practically important margin.To better exploit the precision of systematic sampling we assess the performance of five estimators of variance,including the quasi default.In this study,simulated systematic sampling was applied to artificial populations with contrasting covariance structures and with or without linear trends.We compared the results obtained with the SRS,Matern’s,successive difference replication,Ripley’s,and D’Orazio’s variance estimators.Results:The variances obtained with the four alternatives to the SRS estimator of variance were strongly correlated,and in all study settings consistently closer to the target design variance than the estimator for SRS.The latter always produced the greatest overestimation.In populations with a near zero spatial autocorrelation,all estimators,performed equally,and delivered estimates close to the actual design variance.Conclusion:Without a linear trend,the SDR and DOR estimators were best with variance estimates more narrowly distributed around the benchmark;yet in terms of the least average absolute deviation,Matern’s estimator held a narrow lead.With a strong or moderate linear trend,Matern’s estimator is choice.In large populations,and a low sampling intensity,the performance of the investigated estimators becomes more similar.展开更多
基金supported by grants from the National Natural Sciences Foundation of China (No. 30200284, No. 30600278, No. 30772359)Program for New Century Excellent Talents in University (NCET-06-0641)Scientific Research Foundation for the Returned Overseas Chinese Scholars (2008889)
文摘The optimal antisense accessible sites （AAS） of uroplakin Ⅱ （UP Ⅱ ） mRNA, a specific gene expressed in bladder urothelium, were selected in order to provide a novel method for targeted therapy of transitional cell carcinoma （TCC） of bladder. The 20 mer random oligonucleotide library was synthesized, hybridized with in vitro transcripted total UPⅡ cRNA, then digested by RNase H. After primer extension and autoradiography, the AAS of UPⅡ were selected. The RNADraw soft- ware was used to analyze and choose the AAS with obvious stem-loop structures, according to which the complementary antisense oligonucleotides （AS-ODN） were synthesized. With the AS-ODN0 designed only by RNADraw as a control, the AS-ODNs were transferred into UP Ⅱ highly-expressing cell line RT4. The cellular expression of UPⅡ mRNA was detected by RT-PCR and Western blot. Twelve AAS of UPⅡ mRNA were selected in vitro. Four AAS with stem-loop structures were chosen, locating at 558-577, 552-571, 217-236 and 97-116 bp of UP Ⅱ mRNA respectively. After transfection with the corresponding AS-ODN （AS-ODN1, AS-ODN2, AS-ODN3 and AS-ODN4） for 18 h, the UPⅡ mRNA levels in RT4cells were reduced by 29,3%, 82.7%, 71.3% and 70.9%, while UP Ⅱ protein levels were decreased by 20.2%, 78.5%, 65.2% and 64.4% respectively, which were significantly higher than those of AS-ODN0 （14.3%, 12.1% respectively） （P〈0.01）. The AAS of UPⅡ mRNA was effectively selected in vitro by random oligonucletide library/RNase H cleavage method in combination with computer software analysis, which had important reference values for further studying biological functions of UP Ⅱ gene and targeted therapeutic strategy for TCC of bladder.
文摘Building codes have widely considered the shear wave velocity to make a reliable subsoil seismic classification, based on the knowledge of the mechanical properties of material deposits down to bedrock. This approach has limitations because geophysical data are often very expensive to obtain. Recently, other alternatives have been proposed based on measurements of background noise and estimation of the H/V amplification curve. However, the use of this technique needs a regulatory framework before it can become a realistic site classification procedure. This paper proposes a new formulation for characterizing design sites in accordance with the Algerian seismic building code (RPA99/ver.2003), through transfer functions, by following a stochastic approach combined to a statistical study. For each soil type, the deterministic calculation of the average transfer function is performed over a wide sample of 1-D soil profiles, where the average shear wave (S-W) velocity, V<sub>s</sub>, in soil layers is simulated using random field theory. Average transfer functions are also used to calculate average site factors and normalized acceleration response spectra to highlight the amplification potential of each site type, since frequency content of the transfer function is significantly similar to that of the H/V amplification curve. Comparison is done with the RPA99/ver.2003 and Eurocode8 (EC8) design response spectra, respectively. In the absence of geophysical data, the proposed classification approach together with micro-tremor measures can be used toward a better soil classification.
文摘目的采用两样本孟德尔随机化(MR)探讨417种循环代谢产物与吉兰-巴雷综合征(GBS)风险的因果关系。方法通过MRC IEU OpenGWAS项目获得3个循环代谢产物全基因组关联研究(GWAS)数据,分别为“met-a”“met-c”和“met-d”。GBS相关的单核苷酸多态性位点(SNPs)数据来源于芬兰生物银行数据库,表型代码为“finn-b-G6_GUILBAR”。将来自GWAS的与循环代谢产物密切相关的遗传变异数据(SNPs)作为工具变量(IVs),与来自芬兰的GBS GWAS数据进行双样本MR分析,主要采用随机效应模型的逆方差加权(IVW)方法,根据效应指标优势比(OR)和95%CI评估结果。使用留一法、异质性检验、水平基因多效性检验验证结果的稳定性和可靠性。结果共5种循环代谢产物具有与GBS因果关系的提示性证据。其中,肌酐(OR=2.924,95%CI:1.194~7.163,P=0.019)、谷氨酰胺(OR=1.902,95%CI:1.007~3.592,P=0.048)、异戊酰基肉碱(OR=140.767,95%CI:3.510~5645.336,P=0.009)的循环水平与GBS风险较高有关。相反,基因预测葡萄糖(OR=0.308,95%CI:0.010~0.981,P=0.046)、X-11491(OR=0.069,95%CI:0.007~0.707,P=0.024)的循环水平与GBS风险呈负相关。结论肌酐、谷氨酰胺、异戊酰基肉碱、葡萄糖、X-11491可能与GBS有因果关系。
文摘α-Amylase is widely used in biotechnology industries such as food,textile and detergent.It occupies an important position in the domestic and international enzyme market.An important reason that amylase can achieve its function is it has catalysis domain,which is a very basilic domain,and the function of the catalysis domain mainly depends on catalytic active site.So changing the base of the active site or the base around the active site may affect a lot on α-amylase.This article mainly describes using random mutagenesis of the α-amylase by error-prone PCR,detecting the amylase activity of the mutant strains,and then sequencing the mutants.8 available mutants was picked to do further analysis and prediction of 3D structure.Choosing the mutant with the highest enzyme activity improvement as a further research target molecule,the potential structure changes were predicted by replacement of amino acid residue at the mutant site with other 18 residues.
文摘Background:Large area forest inventories often use regular grids(with a single random start)of sample locations to ensure a uniform sampling intensity across the space of the surveyed populations.A design-unbiased estimator of variance does not exist for this design.Oftentimes,a quasi-default estimator applicable to simple random sampling(SRS)is used,even if it carries with it the likely risk of overestimating the variance by a practically important margin.To better exploit the precision of systematic sampling we assess the performance of five estimators of variance,including the quasi default.In this study,simulated systematic sampling was applied to artificial populations with contrasting covariance structures and with or without linear trends.We compared the results obtained with the SRS,Matern’s,successive difference replication,Ripley’s,and D’Orazio’s variance estimators.Results:The variances obtained with the four alternatives to the SRS estimator of variance were strongly correlated,and in all study settings consistently closer to the target design variance than the estimator for SRS.The latter always produced the greatest overestimation.In populations with a near zero spatial autocorrelation,all estimators,performed equally,and delivered estimates close to the actual design variance.Conclusion:Without a linear trend,the SDR and DOR estimators were best with variance estimates more narrowly distributed around the benchmark;yet in terms of the least average absolute deviation,Matern’s estimator held a narrow lead.With a strong or moderate linear trend,Matern’s estimator is choice.In large populations,and a low sampling intensity,the performance of the investigated estimators becomes more similar.
