AIM: To select the optimal antisense accessible sites of survivin, a highly expressed gene in tumor tissues, in order to explore a novel approach to improve biological therapy of gastric cancer.METHODS: The 20 mer ran...AIM: To select the optimal antisense accessible sites of survivin, a highly expressed gene in tumor tissues, in order to explore a novel approach to improve biological therapy of gastric cancer.METHODS: The 20 mer random oligonucleotide library was synthesized, hybridized with in vitro transcribed total survivin cRNA, then digested by RNase H. After primer extension and autoradiography, the antisense accessible sites (AAS) of survivin were selected. Then RNADraw software was used to analyze and choose the AAS with obvious stem-loop structures, according to which the complementary antisense oligonucleotides (AS-ODNs) were synthesized and transferred into survivin highly- expressing gastric cancer cell line MKN-45. Survivin expression was detected by RT-PCR and Western Blotting. Cellular growth activities were assayed by tetrazolium bromide (MTT)colorimetry. Cellular ultrastructure was observed by electronic microscopy, while apoptosis was detected by annexin V-FITC and propidium iodide staining flow cytometry.RESULTS: Thirteen AAS of survivin were selected in vitro.Four AAS with stem-loop structures were chosen, locating at 207-226 bp, 187-206 bp, 126-145 bp and 44-63 bp of survivin cDNA respectively. When compared with nontranfection controls, their corresponding AS-ODNs (AS-ODN1,AS-ODN2, AS-ODN3 and AS-ODN4) could reduce Survivin mRNA levels in MKN-45 cells by 54.3±1.1% (t= 6.12, P＜0.01),86.1±1.0% (t= 5.27, P＜0.01), 32.2±1.3% (t= 7.34, P＜0.01)and 56.2±0.9% (t= 6.45, P＜0.01) respectively, while survivin protein levels were decreased by 42.2±2.5% (t = 6.26,P＜0.01), 75.4±3.1% (t= 7.11, P＜0.01), 28.3±2.0% (t= 6.04,P＜0.01) and 45.8±1.2% (t = 6.38, P＜0.01) respectively.After transfection with 600 nmol/L AS-ODN1～AS-ODN4 for24 h, cell growth was inhibited by 28.12±1.54% (t= 7.62,P＜0.01), 38.42±3.12% (t = 7.75, P＜0.01), 21.46±2.63%(t= 5.94, P＜0.01) and 32.12±1.77% (t= 6.17, P＜0.01)respectively. Partial cancer cells presented the characteristic morphological changes of apoptosis, with apoptotic rates being 19.31±1.16% (t=7.16, P＜0.01), 29.24±1.94%(t = 8.15, P＜0.01), 11.87±0.68% (t= 6.68, P＜0.01) and21.68±2.14% (t = 7.53, P＜0.01) respectively.CONCLUSION: The AAS of survivin could be effectively selected in vitro by random oligonucleotide library/RNase H cleavage method combined with computer software analysis, this has important reference values for further studying survivin-targeted therapy strategies for gastric cancer.展开更多
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
在FactoryTalkViewSiteEdition中,数据通常存储在内部的文件集中,无法打开此文件集查看数据,也无法对数据进行处理和运算。本文利用开放式数据库连接(OpenDatabaseConnection,ODBC)数据源与微软公司的推出的Access数据库进行连接,将想...在FactoryTalkViewSiteEdition中,数据通常存储在内部的文件集中,无法打开此文件集查看数据,也无法对数据进行处理和运算。本文利用开放式数据库连接(OpenDatabaseConnection,ODBC)数据源与微软公司的推出的Access数据库进行连接,将想要处理的数据记录到Access数据库中,并可以将此结果在FactoryTalkView Site Edition中的趋势图中显示,另外后期可以将想要的数据从Access数据库中提取出来进行进一步处理和运算。展开更多
AIM:To review an evolution of laparoscopic surgery,there has been a growing interest in laparoendoscopic single-site surgery(LESS).METHODS:A comprehensive electronic literature search was conducted using PubM ed datab...AIM:To review an evolution of laparoscopic surgery,there has been a growing interest in laparoendoscopic single-site surgery(LESS).METHODS:A comprehensive electronic literature search was conducted using PubM ed database to identify all publications relating to LESS-partial nephrectomy(PN).The research includes articles published from April 2008 to January 2014.We focused our attention only on articles in which were cited the single-site surgical technique(laparoscopic and robotic),tumour stage and grade,mean tumour size,intraoperative variables,blood loss and transfusion rate,length of postoperative stay and complication rates,Clavien classification,positive of surgical margins,pain assessment at discharge.RESULTS:A total of 9 studies were collected with 221patients included.The mean patients age was 62 years.The mean tumor size was 2.35 cm with a mean operative time of 181 min(range 111-270 min)and 58.3%were done by robot.The mean ischemia time was 23.6min.The 25.8%of patients underwent an unclamp LESS-PN.Mean estimated blood loss was 296 mL and median length of hospital stay was 4 d.The rate of severe post-operative complications(≥Clavien gradeⅢ)was 5.4%.Not all surgical series of LESS-PN or Robotic-LESS-PN shows conversion in Multiport Laparoscopic or Open Surgery.Regarding oncologic outcomes,surgical margins were positive 4%of patients(9/221),no distant or port-site metastases were recorded.CONCLUSION:LESS-PN and RLESS-PN are feasible and associated with reduced postoperative pain,shorter median hospital stay,shorter recovery time,and better cosmetic satisfaction without compromising surgical and oncological safety.展开更多
Alumina(Al_(2)O_(3))is widely used in the chemical industry as the catalyst and support due to its high specific surface area,abundant pore size distribution and chemical stability.However,the occurrence of hydration ...Alumina(Al_(2)O_(3))is widely used in the chemical industry as the catalyst and support due to its high specific surface area,abundant pore size distribution and chemical stability.However,the occurrence of hydration in water environment,result in outstanding decrease in specific surface area and collapse of pore structure.In this work,dodecyl phosphoric acid(PA)is used to modify the surface of Al_(2)O_(3)to obtain a series of hydrophobic material(Al_(2)O_(3)-PA).Based on XPS and NMR analysis,PA is chemically bonded on Al_(2)O_(3)to form PAOAAl bond.Furthermore,BET and WCA results display that Al_(2)O_(3)-1PA exhibits excellent the hydrophobicity and hydrothermal stability while maintains the pore structure.Take it as the substrate to support the Pd nanoparticles,the as-prepared Pd/Al_(2)O_(3)-PA shows the superior catalytic performance in the hydrogenation of phenol and anthraquinone relative to Pd/Al_(2)O_(3),indicating the accessibility of Pd sites after PA modification.Especially,the significantly enhanced stability is also obtained in four cycles for aqueous phenol hydrogenation.This can be ascribed that the PA modification inhibits the aggregation of Pd nanoparticles and the products adhesion in the reaction process.The extension of PA coatings to monolithic catalysts could expand their current capabilities in industrial applications and warrants ongoing investigation.展开更多
基金Supported by National Natural Science Foundation of China, No.30200284Science Foundation of Huazhong University of Science and Technology
文摘AIM: To select the optimal antisense accessible sites of survivin, a highly expressed gene in tumor tissues, in order to explore a novel approach to improve biological therapy of gastric cancer.METHODS: The 20 mer random oligonucleotide library was synthesized, hybridized with in vitro transcribed total survivin cRNA, then digested by RNase H. After primer extension and autoradiography, the antisense accessible sites (AAS) of survivin were selected. Then RNADraw software was used to analyze and choose the AAS with obvious stem-loop structures, according to which the complementary antisense oligonucleotides (AS-ODNs) were synthesized and transferred into survivin highly- expressing gastric cancer cell line MKN-45. Survivin expression was detected by RT-PCR and Western Blotting. Cellular growth activities were assayed by tetrazolium bromide (MTT)colorimetry. Cellular ultrastructure was observed by electronic microscopy, while apoptosis was detected by annexin V-FITC and propidium iodide staining flow cytometry.RESULTS: Thirteen AAS of survivin were selected in vitro.Four AAS with stem-loop structures were chosen, locating at 207-226 bp, 187-206 bp, 126-145 bp and 44-63 bp of survivin cDNA respectively. When compared with nontranfection controls, their corresponding AS-ODNs (AS-ODN1,AS-ODN2, AS-ODN3 and AS-ODN4) could reduce Survivin mRNA levels in MKN-45 cells by 54.3±1.1% (t= 6.12, P＜0.01),86.1±1.0% (t= 5.27, P＜0.01), 32.2±1.3% (t= 7.34, P＜0.01)and 56.2±0.9% (t= 6.45, P＜0.01) respectively, while survivin protein levels were decreased by 42.2±2.5% (t = 6.26,P＜0.01), 75.4±3.1% (t= 7.11, P＜0.01), 28.3±2.0% (t= 6.04,P＜0.01) and 45.8±1.2% (t = 6.38, P＜0.01) respectively.After transfection with 600 nmol/L AS-ODN1～AS-ODN4 for24 h, cell growth was inhibited by 28.12±1.54% (t= 7.62,P＜0.01), 38.42±3.12% (t = 7.75, P＜0.01), 21.46±2.63%(t= 5.94, P＜0.01) and 32.12±1.77% (t= 6.17, P＜0.01)respectively. Partial cancer cells presented the characteristic morphological changes of apoptosis, with apoptotic rates being 19.31±1.16% (t=7.16, P＜0.01), 29.24±1.94%(t = 8.15, P＜0.01), 11.87±0.68% (t= 6.68, P＜0.01) and21.68±2.14% (t = 7.53, P＜0.01) respectively.CONCLUSION: The AAS of survivin could be effectively selected in vitro by random oligonucleotide library/RNase H cleavage method combined with computer software analysis, this has important reference values for further studying survivin-targeted therapy strategies for gastric cancer.
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.
文摘在FactoryTalkViewSiteEdition中,数据通常存储在内部的文件集中,无法打开此文件集查看数据,也无法对数据进行处理和运算。本文利用开放式数据库连接(OpenDatabaseConnection,ODBC)数据源与微软公司的推出的Access数据库进行连接,将想要处理的数据记录到Access数据库中,并可以将此结果在FactoryTalkView Site Edition中的趋势图中显示,另外后期可以将想要的数据从Access数据库中提取出来进行进一步处理和运算。
文摘AIM:To review an evolution of laparoscopic surgery,there has been a growing interest in laparoendoscopic single-site surgery(LESS).METHODS:A comprehensive electronic literature search was conducted using PubM ed database to identify all publications relating to LESS-partial nephrectomy(PN).The research includes articles published from April 2008 to January 2014.We focused our attention only on articles in which were cited the single-site surgical technique(laparoscopic and robotic),tumour stage and grade,mean tumour size,intraoperative variables,blood loss and transfusion rate,length of postoperative stay and complication rates,Clavien classification,positive of surgical margins,pain assessment at discharge.RESULTS:A total of 9 studies were collected with 221patients included.The mean patients age was 62 years.The mean tumor size was 2.35 cm with a mean operative time of 181 min(range 111-270 min)and 58.3%were done by robot.The mean ischemia time was 23.6min.The 25.8%of patients underwent an unclamp LESS-PN.Mean estimated blood loss was 296 mL and median length of hospital stay was 4 d.The rate of severe post-operative complications(≥Clavien gradeⅢ)was 5.4%.Not all surgical series of LESS-PN or Robotic-LESS-PN shows conversion in Multiport Laparoscopic or Open Surgery.Regarding oncologic outcomes,surgical margins were positive 4%of patients(9/221),no distant or port-site metastases were recorded.CONCLUSION:LESS-PN and RLESS-PN are feasible and associated with reduced postoperative pain,shorter median hospital stay,shorter recovery time,and better cosmetic satisfaction without compromising surgical and oncological safety.
基金supported by National Key Research&Development Program of China(2021YFB3801600)Fundamental Research Funds for the Central University(buctrc201921,JD2223,12060093063)Innovative Achievement Commercialization Service-Platform of Industrial Catalysis(2019-00900-2-1).
文摘Alumina(Al_(2)O_(3))is widely used in the chemical industry as the catalyst and support due to its high specific surface area,abundant pore size distribution and chemical stability.However,the occurrence of hydration in water environment,result in outstanding decrease in specific surface area and collapse of pore structure.In this work,dodecyl phosphoric acid(PA)is used to modify the surface of Al_(2)O_(3)to obtain a series of hydrophobic material(Al_(2)O_(3)-PA).Based on XPS and NMR analysis,PA is chemically bonded on Al_(2)O_(3)to form PAOAAl bond.Furthermore,BET and WCA results display that Al_(2)O_(3)-1PA exhibits excellent the hydrophobicity and hydrothermal stability while maintains the pore structure.Take it as the substrate to support the Pd nanoparticles,the as-prepared Pd/Al_(2)O_(3)-PA shows the superior catalytic performance in the hydrogenation of phenol and anthraquinone relative to Pd/Al_(2)O_(3),indicating the accessibility of Pd sites after PA modification.Especially,the significantly enhanced stability is also obtained in four cycles for aqueous phenol hydrogenation.This can be ascribed that the PA modification inhibits the aggregation of Pd nanoparticles and the products adhesion in the reaction process.The extension of PA coatings to monolithic catalysts could expand their current capabilities in industrial applications and warrants ongoing investigation.
