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
Capital Normal University Library(CNU Library) initiated the first practical application of a 3D virtual library sites-navigation system(an electronic kiosk version)among Chinese academic and research libraries in 201...Capital Normal University Library(CNU Library) initiated the first practical application of a 3D virtual library sites-navigation system(an electronic kiosk version)among Chinese academic and research libraries in 2010. It was primarily based on the technologies of 3DsMax and Virtools. This paper concentrates on the discussion of the methods in creating the 3D model and in realizing the interaction among the data usage of the system. As a result, several important service functions of the system have been developed successfully so far for convenient public access. They include the functions of virtual-book searching, path navigation online, real-time message exchanges, and multi-media sharing, etc.展开更多
In the context of constructing national public cultural demonstration areas all over China and taking the system innovation history of Baoji as an example,this paper studies the path choices and service system design ...In the context of constructing national public cultural demonstration areas all over China and taking the system innovation history of Baoji as an example,this paper studies the path choices and service system design of the development of service system of public libraries in the western region of China.The results show that the key issue the China public library service system is facing is to promote system innovation;the 'chain system and grassroots sites of libraries' is a new synthesized innovative program on public library service system in accordance with the actual situations of Baoji area and other areas in the western region of China;the service system design of local public libraries should insist on seeking truth from facts,and carrying out an integrated,simple and feasible reform according to local conditions.展开更多
从水生栖热菌(Thermus aquaticus)YT-1中分离得到的Taq DNA 聚合酶是一种广泛应用于PCR的耐热DNA聚合酶。由于天然菌株酶产量较低,培养条件要求严格,酶的纯化过程极为繁琐而使产品成本较高,因而促使人们构建适合于大规模生产的基因工程...从水生栖热菌(Thermus aquaticus)YT-1中分离得到的Taq DNA 聚合酶是一种广泛应用于PCR的耐热DNA聚合酶。由于天然菌株酶产量较低,培养条件要求严格,酶的纯化过程极为繁琐而使产品成本较高,因而促使人们构建适合于大规模生产的基因工程菌株。已有人分别通过不同的途径,使用不同的载体,成功地在大肠杆菌菌株中表达了Taq 耐热DNA聚合酶基因。我们通过与前人不同的途径,把这一基因克隆到大肠杆菌的载体质粒pJLA503上并使其得以高表达,为降低生产成本和进一步研究该酶的各种特性提供了有利条件。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.
基金supported by the Beijing Social Science Foundation in China(Grant No.SM201010028014)
文摘Capital Normal University Library(CNU Library) initiated the first practical application of a 3D virtual library sites-navigation system(an electronic kiosk version)among Chinese academic and research libraries in 2010. It was primarily based on the technologies of 3DsMax and Virtools. This paper concentrates on the discussion of the methods in creating the 3D model and in realizing the interaction among the data usage of the system. As a result, several important service functions of the system have been developed successfully so far for convenient public access. They include the functions of virtual-book searching, path navigation online, real-time message exchanges, and multi-media sharing, etc.
基金a periodical research achievement of"Research on the Construction of the System of Main-branch Public Libraries(Sites)"(No.38,Baoji Official Announcement in 2011),which is a project of system design of Baoji creating a demonstration area of national public cultural service system"Research of Free Access to Public Libraries in Shaanxi Province in the Perspective of Public Cultural Space"(No.2012C019),which is a research project of major issues in theory and practice of the circle of social sciences in Shaanxi Province"Research of the Construction of Library Union in Xi’an City"(No.12WL26),which is a project of the fund of planning of social sciences in Xi’an City
文摘In the context of constructing national public cultural demonstration areas all over China and taking the system innovation history of Baoji as an example,this paper studies the path choices and service system design of the development of service system of public libraries in the western region of China.The results show that the key issue the China public library service system is facing is to promote system innovation;the 'chain system and grassroots sites of libraries' is a new synthesized innovative program on public library service system in accordance with the actual situations of Baoji area and other areas in the western region of China;the service system design of local public libraries should insist on seeking truth from facts,and carrying out an integrated,simple and feasible reform according to local conditions.
文摘从水生栖热菌(Thermus aquaticus)YT-1中分离得到的Taq DNA 聚合酶是一种广泛应用于PCR的耐热DNA聚合酶。由于天然菌株酶产量较低,培养条件要求严格,酶的纯化过程极为繁琐而使产品成本较高,因而促使人们构建适合于大规模生产的基因工程菌株。已有人分别通过不同的途径,使用不同的载体,成功地在大肠杆菌菌株中表达了Taq 耐热DNA聚合酶基因。我们通过与前人不同的途径,把这一基因克隆到大肠杆菌的载体质粒pJLA503上并使其得以高表达,为降低生产成本和进一步研究该酶的各种特性提供了有利条件。
