寡核苷酸类

摘要： 目的 研究结核分枝杆菌(MTB)不同基因型在广东省各地区的流行情况及其耐药相关性.方法 随机数字法抽取广州市胸科医院2014-2015年临床分离的来自于广东省不同地区的504株结核分枝杆菌菌株,采用比例法进行药物敏感性实验,选用间隔区寡核苷酸分型方法(Spoligotyping)对结核分枝杆菌进行基因分型.基因型耐药率差异比较采用χ2检验.结果 (1)在504株结核分枝杆菌临床分离株中北京家族和非北京家族分别是386株和118株.非北京家族中,T家族高达48株;其余是H3家族、EAI家族、MANU家族、AMBIGO家族、LAM家族以及新发基因型.北京家族构成比在性别、年龄上无统计学差异(P>0.05),但在汕头、汕尾、揭阳、东莞四个地级市的构成比低于广州市,差异有统计学意义(P0.05).结论 广东省结核分枝杆菌呈基因多态性,北京基因型菌株为本省结核分枝杆菌主要流行株,而且与异烟肼、吡嗪酰胺、阿米卡星耐药相关.非北京基因型在潮汕和东莞等华侨之乡比例有所增加.

摘要： 指数富集的配基数系统进化(SELEX)技术的基本过程是用化学方法合成一个单链寡核苷酸库,将之于与靶标物质混合,寡核苷酸链与靶标物质结合形成复合物,去除未结合的核酸链后,再将结合的核酸分子与靶标物质分离,以此寡核苷酸分子为模板进行PCR扩增,用其产物进行下一个循环的筛选.经过数次循环,即可得到能与靶标物质高亲和力高特异性结合的核酸适配体分子.本文综述了SELEX技术筛选核酸适配体技术的一些研究进展.

摘要： Objective To evaluate the effect of anti-myosin monoclonal antibodies-nuclear factor kappa B (NF-κB) decoy oligodeoxynucleotides-lipofectamine compound (mAb2G4-ODN-lip) on myocardial ischemia-reperfusion (I/R) injury in rats.Methods Forty clean-grade healthy adult male Sprague-Dawley rats,aged 8-10 weeks,weighing 240-260 g,were divided into 4 groups (n=10 each) using a random number table:sham operation group (group S),myocardial I/R group (group I/R),ODN-lip group (group ODN) and mAb2G4-ODN-lip group (group mAb2G4).Myocardial I/R was induced by occlusion of the left anterior descending branch of coronary artery for 30 min followed by 120 min reperfusion.In ODN and mAb2G4 groups,ODN-lip (100 μg ODN) and mAb2G4-ODN-lip (100 μg ODN) compounds were injected via the femoral vein,respectively,immediately after onset of ischemia.The left anterior descending branch of coronary artery was only occluded but not ligated in group S.The animals were sacrificed at 120 min of reperfusion and myocardial specimens of the left ventricle on the ischemic side were obtained for examination of the pathological changes (using haematoxylin and eosin staining) and for determination of the expression of NF-κB (by Western blot) and contents of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) (by enzyme-linked immunosorbent assay).Results Compared with group S,the expression of NF-κB was significantly up-regulated,and the contents of TNF-α and IL-6 were increased in I/R,ODN and mAb2G4 groups (P＜ 0.05).Compared with group I/R,the expression of NF-κB was significantly down-regulated,and the contents of TNF-α and IL-6 were decreased in ODN and mAb2G4 groups (P＜0.05).Compared with group ODN,the expression of NF-κB was significantly down-regulated,and the contents of TNF-α and IL-6 were decreased in group mAb2G4 (P＜0.05).The pathological changes of myocardial tissues were significantly attenuated in group I/R,group ODN and group mAb2G4 in turn.Conclusion mAb2G4-ODN-lip can mitigate myocardial I/R injury in rats.%目的 评价抗肌球蛋白单克隆抗体-NF-κB诱骗性寡核苷酸-阳离子脂质体复合物(mAb2G4-ODN-lip)对大鼠心肌缺血再灌注损伤的影响.方法 清洁级健康成年雄性SD大鼠40只,8～10周龄,体重240～260 g,采用随机数字表法分为4组(n=10):假手术组(S组)、心肌缺血再灌注组(I/R组)、ODN-lip组(ODN组)和mAb2G4-ODN-lip组(mAb2G4组).采用结扎冠状动脉左前降支30 min,恢复灌注120 min的方法制备大鼠心肌缺血再灌注损伤模型.ODN组和mAb2G4组于缺血即刻股静脉注射ODN-lip(100 μg ODN)和mAb2G4-ODN-lip(100 μg ODN)复合物,S组仅穿线,不结扎.于再灌注120 min时处死大鼠,取左心室缺血心肌,HE染色观察心肌组织病理学结果,采用Western blot法检测NF-κB表达;ELISA法检测TNF-α和IL-6含量.结果 与S组相比,I/R组、ODN组和mAb2G4组心肌NF-κB表达上调,TNF-α和IL-6含量升高(P＜0.05);与I/R组相比,ODN组和mAb2G4组心肌NF-κB表达下调,TNF-α和IL-6含量降低(P＜0.05);与ODN组相比,mAb2G4组心肌NF-κB表达下调,TNF-α和IL-6含量降低(P＜0.05).I/R组、ODN组和mAb2G4组心肌组织病理学损伤依次减轻.结论 mAb2G4-ODN-lip可减轻大鼠心肌缺血再灌注损伤.

摘要： Objective To investigate the effect of bcl-XL antisense oligonucleotide transfection on the proliferation of colon cancer cells and the growth of human colon cancer xenografts in nude mice.Methods A long-term SW620 single cell suspension was prepared for subcutaneous injection in nude mice.Nude mice were randomly divided into 3 groups: control group,N-ODN group and antisense oligonucleotide group (n=5),with an average of 50 mm3.The antisense oligonucleotide group and N-ODN group were injected with 2 ml/kg antisense oligonucleotide or N-ODN,and the control group was injected with equal amount of normal saline.At 16th day after the operation,the tumor cells were taken out,and some tumor tissues were extracted.RNA and protein were extracted.The cationic liposome mediated transfection of antisense oligonucleotide,real-time quantitative polymerase chain reaction (Real-time PCR),Western blotting,methyl thiazol tetrazolium (MTT) method and TdT-mediated dUTP nick end labeling (TUNEL) method were applied to observe the effects of bcl-XL antisense oligonucleotide on the proliferation and apoptosis of colorectal cancer cells in vivo and the growth of human colon cancer xenografts.Results (1) The inhibition rate of cell proliferation activity was gradually increased with the increase of antisense oligonucleotide concentration.When the concentration was over 200 nmol/L,the inhibition rate of cell proliferation was significantly decreased with the difference being statistically significant (t=2.418,1.531 and 1.838,P=0.003,0.016 and 0.011).(2) The relative expression levels in the control group,blank control group,N-ODN group and mRNA antisense oligonucleotides group were 0.96±0.25,0.97±0.48,0.78±0.32 and 0.41±0.13,the inhibition rate was 0.00%,-6.25%,18.75% and 57.29%,respectively.There was significant difference between antisense oligonucleotide group and control group,blank control group or N-ODN group (P=0.025).(3) Compared with the control group,there was no significant change in the blank control group;the N-ODN group was slightly lower than that in the control group;the antisense oligonucleotide group was significantly lower (P=0.017).(4) The positive rates of cells in control group,blank control group,N-ODN group and antisense oligonucleotide group were 3.7%,5.0%,7.0% and 35.0%,respectively.By statistical analysis,antisense oligonucleotide group and cell control group,blank control group and N-ODN group,the difference was statistically significant (χ2=181.880,P=0.000);cells in the control group,N-ODN group and control group comparison between 22 blank,there was no statistically significant difference (χ2=1.410,P=0.084).(5) There was no significant difference in tumor volume between first groups (P=0.162).On the sixth day,there was significant difference between the antisense oligonucleotide group and the control group (t=2.410,P=0.027).On the eleventh and the 16 day,there was significant difference between the two groups in (P=0.028,0.016).(6) The expression of bcl-XL gene in the tumor tissues of the 3 groups of nude mice was significantly decreased after antisense oligonucleotide treatment,but the expression of antisense oligonucleotide group was significantly decreased,while the expression of N-ODN in the control group and the control group did not change significantly.Conclusion bcl-XL antisense oligonucleotides can effectively inhibit the proliferation of colon cancer cells and the growth of human colon cancer xenografts in nude mice.To reduce the expression of bcl-XL by antisense oligonucleotides,and to provide the experimental basis for gene therapy of colon cancer.%目的 观察bcl-XL反义寡核苷酸转染后,对结肠癌细胞增殖及裸鼠体内人结肠癌移植瘤生长的影响.方法 制备对数生长期的SW620单细胞悬液,裸鼠背侧皮下注射.当移植瘤体积平均50mm时,将裸鼠随机分为3组:对照组、N-ODN组和反义寡核苷酸组,每组5只.反义寡核苷酸组及N-ODN组瘤旁注射2ml/kg体重的反义寡核苷酸或N-ODN,对照组瘤旁注射等量生理盐水,第16天处死裸鼠,取部分肿瘤组织,提取RNA及蛋白,采用阳离子脂质体介导的反义寡核苷酸转染、实时定量聚合酶链反应(Real-time PCR)、Western blot、噻唑蓝(MTT)法、原位缺口末端标记法(TUNEL)等方法观察了bcl-XL反义寡核苷酸转染对结肠癌细胞增殖、凋亡的影响及裸鼠体内人结肠癌移植瘤生长的影响.结果 (1)对细胞增殖活性的抑制率随反义寡核苷酸浓度的增加逐渐增高,具剂量依赖性.当浓度超过200nμmol/L后,对细胞活性的增殖抑制率递增幅度明显减小,差异有统计学意义(t=2.418、1.531和1.838,P=0.003、0.016和0.011).(2)细胞对照组、空白对照组、N-ODN组及反义寡核苷酸组的mRNA相对表达量分别为0.96±0.25、0.97±0.48、0.78±0.32及0.41±0.13,抑制率分别为0.00%、-6.25%、18.75%及57.29%,与细胞对照组比较,除空白对照组有少量增高外,N-ODN组及反义寡核苷酸组的表达依次减弱,抑制率依次增加.反义寡核苷酸组与细胞对照组、空白对照组及N-ODN组比较,差异有统计学意义(P=0.025);N-ODN组、空白对照组及细胞对照组间两两比较,差异无统计学意义(P=0.263).(3)与细胞对照组比较,空白对照组无明显变化;N-ODN组稍有降低;反义寡核苷酸组明显降低(P=0.017).(4)细胞对照组、空白对照组、N-ODN组及反义寡核苷酸组的阳性细胞的阳性率分别为3.7%、5.0%、7.0%及35.0%.经统计学分析,反义寡核苷酸组与细胞对照组、空白对照组及N-ODN组比较,差异有统计学意义(χ2=181.880,P=0.000);细胞对照组、空白对照组及N-ODN组间两两比较,差异无统计学意义(χ2=1.410,P=0.084).(5)治疗第1天,肿瘤体积各组间差异无统计学意义(P=0.162).第6天,反义寡核苷酸组与对照组比较差异有统计学意义(t=2.410,P=0.027).第11、16天,各组两两比较差异均有统计学意义(P=0.028、0.016).(6)反义寡核甘酸体内治疗后,3组裸鼠肿瘤组织中均有bcl-XL基因的表达,但反义寡核甘酸组的表达明显减弱,而对照组及N-ODN组的表达未发生明显改变.结论 bcl-XL反义寡核苷酸可有效抑制结肠癌细胞增殖及裸鼠体内人结肠癌移植瘤的生长.通过反义寡核苷酸下调bcl-XL的表达,可治疗结肠癌.

摘要： 目的:研究不同端粒寡核苷酸序列对大鼠胸大动脉平滑肌细胞(A7R5)凋亡的影响,及沉默信息调节因子1(SIRT1)/抑癌基因P53(P53)信号通路是否参与调控端粒寡核苷酸序列诱导A7R5的凋亡.方法:将三种端粒重复序列端粒寡核苷酸序列的二聚体、四聚体、六聚体[TE-12(TTAGGG)2、TE-24(TTAGGG)4、TE-36(TTAGGG)6]转染至A7R5,分别作为TE-12组、TE-24组、TE-36组,另设A7R5空白为阴性对照组.应用流式细胞仪检测各端粒寡核苷酸序列转染后A7R5的凋亡情况.采用逆转录多聚酶联反应(RT-PCR)及蛋白免疫印迹(Western-blot)检测TE-12组、TE-24组、TE-36组及阴性对照组SIRT1、P53基因及蛋白水平.结果:将TE-12、TE-24、TE-36成功转染进A7R5,各组转染率差异无统计学意义.TE-12组A7R5凋亡率明显高于TE-36组、TE-24组和阴性对照组,TE-24组细胞凋亡率最低,但亦明显高于阴性对照组,差异均有统计学意义(P均<0.05).TE-12组A7R5SIRT1mRNA及蛋白水平明显高于TE-36组、TE-24组和阴性对照组,差异均有统计学意义(P均<0.05).TE-24组A7R5P53mRNA及蛋白水平明显低于TE-36组、TE-12组和阴性对照组,差异均有统计学意义(P均<0.05).结论:端粒寡核苷酸序列促进A7R5的凋亡.不同结构的端粒寡核苷酸序列引起A7R5凋亡程度不同,在选取的三种结构中,端粒单链序列TE-12引起A7R5凋亡最明显.端粒寡核苷酸序列对A7R5凋亡的作用可能与SIRT1/P53信号通路有关.%Objective: To study the influence of different telomere oligonucleotide on rat's thoracic aortic smooth muscle (A7R5 ) cell apoptosis and to clarify weather silent information regulator 1 (SIRT1)/cancer suppressor gene (P53) signaling pathway involved in oligonucleotide induced cell apoptosis. Methods: 3 telomere repeat sequences of TE-12 (TTAGGG)2, TE-24 (TTAGGG)4 and TE-36 (TTAGGG)6 were respectively transfected into rat's A7R5 cells as TE-12 group, TE-24 group and TE-36 group; in addition, blank A7R5 cells were used as Control group. Transfection induced A7R5 cell apoptosis was detected by flow cytometry, mRNA and protein expressions of SIRT1 and P53 in A7R5 cells were examined by RT-PCR and Western blot analysis in different groups. Results: Successful ransfection rates were similar among 3 groups. Compared with Control group and TE-36, TE-24 groups, the highest apoptosis rate of A7R5 cells was found in TE-12 group and the lowest was found in TE-24 group which was still higher than that in Control group, allP<0.05. mRNA and protein expressions of SIRT1 was obviously higher in TE-12 group than the other 3 groups, allP<0.05; mRNA and protein expressions of P53 was obviously lower in TE-24 group than the other 3 groups, allP<0.05. Conclusion: Telomere oligonucleotide sequence may promote rat's A7R5 cell apoptosis; different sequence had various influence and the strongest effect was observed in TE-12 sequence. The above impact might be related to SIRT1/P53 signaling pathway.

摘要： Objective To investigate the effect of oligodeoxynucleotide mt01 (ODN mt01) on the osteogenic differentiation of human periodontal ligament cells (hPDLCs), and to provide an experimental basis for the develop-ment of clinical agents. Methods The in vitro hPDLCs were included as the subjects in the study for primary culture and subculture. The third generation of hPDLCs and ODN mt01 with the working concentration of 1.0 mg/L were co-cultured for 24 h, 48 h, 72 h and 96 h. The effect of ODN mt01 on the osteogenic differentiation of hPDLCs was de-termined by real time PCR. Results At 24 h, 48 h, 72 h after the co-culture, ODN mt01 with the working concentra-tion of 1.0 mg/L significantly up-regulated the mRNA expression levels of the osteogenic factors Runx-2, Sp7 and Col-lagen-I in hPDLCs, respectively, compared with those in the PBS control group ( P<0.05). Conclusion The appropri-ate working concentration of ODN mt01 may increase the gene expression levels of the osteogenic factors to promote the osteogenic differentiation of hPDLCs.%目的：研究寡核苷酸（ODN）mt01对人牙周膜细胞向成骨细胞分化的影响，为临床药剂的开发奠定实验基础。方法采用人离体牙牙周膜细胞作为研究对象，原代培养牙周膜细胞并传代，取第三代人牙周膜细胞，加入工作浓度为1.0 mg/L的ODN mt01共培养24 h、48 h、72 h和96 h后，通过实时聚合酶链反应（real time PCR）法研究ODN mt01对人牙周膜细胞成骨分化的影响。结果分别作用于人牙周膜细胞24 h、48 h、72 h和96 h后，与磷酸盐缓冲液（PBS）对照组相比，工作浓度为1.0 mg/L的ODN mt01可显著上调人牙周膜细胞中Runx-2、Sp7和Ⅱ型胶原等成骨相关因子mRNA的表达水平（P＜0.05）。结论适当工作浓度的ODN mt01可能通过增加成骨相关因子的基因表达水平进而促进人牙周膜细胞的成骨向分化。

摘要： Objective To investigate the molecular mechanism of low-dose X-ray irradiation on osteoblasts by detecting the gene expression profiles of osteoblasts radiated with 0.5 Gy X-ray.Methods Microarray analyses to value the changes of gene expression of MC3T3-E1 osteoblasts after 0.5 Gy X-ray irradiation were conducted.The end points included modulation of key markers,and pathway and gene ontology through transcriptomic profiling and bioinformatics analysis.Several major genes during osteoblasts differentiation were selected for Real-time PCR analysis.Results 1 412 differentially expressed transcripts were identified between the radiated group and control group.Among the identified genes,559 transcripts were up-regulated and 853 transcripts were down-regulated after irradiation.Real-time PCR analysis indicated that the mRNA expressions of Bglap-rs1,Col1a2,Tgf-β1,Lrp5,Dvl1,Map4k5 were apparently higher than that of the controls (P ＜ 0.01),whereas the Id2 and Dkk1 expression decreased under the same condition (P ＜ 0.01).Western-blot analysis indicated TGF-β1 and Lrp-5 protein expression increased and Dkk1 expression decreased(P ＜ 0.05).Pathway analysis and gene ontology analysis revealed that focal adhesion,extracellular signaling,cytoskeletal protein binding,Wnt signal pathway,IκB/NF-κB signal pathway and growth factor receptors activities.Conclusions Low-dose X-ray radiation promoted osteoblasts differentiation in which focal adhesion,extracellular signaling,cytoskeletal protein binding,Wnt signal pathway,IκB/NF-κB signal pathway and growth factor activities might be involved.%目的 探索低剂量X线促进成骨分化的关分子机制.方法 0.5 Gy X线照射成骨细胞后继续诱导成骨培养,采用基因芯片技术检测X线照射成骨细胞后基因表达谱的变化,筛检出表达差异基因,选取部分骨分化相关基因通过实时聚合酶链反应(real-time PCR)进行验证,并进行基因本体(Gene Ontology,GO)分析和通路(Pathway)分析.结果 成骨细胞在低剂量X射线照射后,共发现1 412个差异表达基因,其中559个上调,853个下调.Real-time PCR结果显示,0.5 Gy X线照射组mRNA表达水平显著升高的基因包括:Bglap-rs1(2.5倍)、Col1 a2(2.1倍)、Tgf-β1(3.4倍)、Lrp5 (2.5倍)、Dvl1(4.1倍)、Map4k5(1.6倍)(P＜0.01),而Id2(0.5倍)、Dkk1(0.2倍)表达水平显著降低(P＜0.01).免疫印迹结果显示0.5 GyX线照射成骨细胞后TGF-β1、LRP-5蛋白表达增加(P＜0.05),Dkk1蛋白表达减低(P＜0.01).GO和Pathway分析结果显示细胞外信号、细胞黏附斑信号、Wnt信号、IκB/NF-κB信号、生长因子受体信号明显富集.结论 低剂量X线照射成骨细胞后,可能通过细胞外信号、细胞黏附斑信号、Wnt信号、IκB/NF-κB信号、生长因子受体信号通路促进成骨细胞分化和矿化,这些信号分子可以相互作用形成复杂的网络体系,最终导致成骨细胞表型特征的改变性.

摘要： 目的:研究端粒酶逆转录酶(TERT)的反义寡核苷酸(ASODN)与甲状腺癌(thyroid carcinoma，TC)细胞凋亡的关系。rn 方法:体外培养TC细胞，TERT ASODN和SODN分别作用于体外培养的TC细胞，采用透射电镜、Hoechest荧光染色、流式细胞仪等方法分析ASODN TERT对TC细胞凋亡的影响。rn 结果:透射电镜示ASODN可诱导TC出现典型的细胞凋亡形态，Hoechest荧光染色显示ASODN可使TC凋亡数目增多从48±19上升到181±28，流式细胞仪证实5 μmol/LASODN处理组、SODN处理组及空白对照组的细胞凋亡率分别为11.2％，2.1％和无凋亡(P＜0.05)。rn 结论:TERT ASODN能增强TC细胞凋亡。

摘要： 目的:研究端粒酶逆转录酶(TERT)的反义寡核苷酸(ASODN)与甲状腺癌(thyroid carcinoma，TC)细胞凋亡的关系。rn 方法:体外培养TC细胞，TERT ASODN和SODN分别作用于体外培养的TC细胞，采用透射电镜、Hoechest荧光染色、流式细胞仪等方法分析ASODN TERT对TC细胞凋亡的影响。rn 结果:透射电镜示ASODN可诱导TC出现典型的细胞凋亡形态，Hoechest荧光染色显示ASODN可使TC凋亡数目增多从48±19上升到181±28，流式细胞仪证实5 μmol/LASODN处理组、SODN处理组及空白对照组的细胞凋亡率分别为11.2％，2.1％和无凋亡(P＜0.05)。rn 结论:TERT ASODN能增强TC细胞凋亡。

摘要： 目的:研究端粒酶逆转录酶(TERT)的反义寡核苷酸(ASODN)与甲状腺癌(thyroid carcinoma，TC)细胞凋亡的关系。rn 方法:体外培养TC细胞，TERT ASODN和SODN分别作用于体外培养的TC细胞，采用透射电镜、Hoechest荧光染色、流式细胞仪等方法分析ASODN TERT对TC细胞凋亡的影响。rn 结果:透射电镜示ASODN可诱导TC出现典型的细胞凋亡形态，Hoechest荧光染色显示ASODN可使TC凋亡数目增多从48±19上升到181±28，流式细胞仪证实5 μmol/LASODN处理组、SODN处理组及空白对照组的细胞凋亡率分别为11.2％，2.1％和无凋亡(P＜0.05)。rn 结论:TERT ASODN能增强TC细胞凋亡。

摘要： 目的:研究端粒酶逆转录酶(TERT)的反义寡核苷酸(ASODN)与甲状腺癌(thyroid carcinoma，TC)细胞凋亡的关系。rn 方法:体外培养TC细胞，TERT ASODN和SODN分别作用于体外培养的TC细胞，采用透射电镜、Hoechest荧光染色、流式细胞仪等方法分析ASODN TERT对TC细胞凋亡的影响。rn 结果:透射电镜示ASODN可诱导TC出现典型的细胞凋亡形态，Hoechest荧光染色显示ASODN可使TC凋亡数目增多从48±19上升到181±28，流式细胞仪证实5 μmol/LASODN处理组、SODN处理组及空白对照组的细胞凋亡率分别为11.2％，2.1％和无凋亡(P＜0.05)。rn 结论:TERT ASODN能增强TC细胞凋亡。

摘要： 目的:研究端粒酶逆转录酶(TERT)的反义寡核苷酸(ASODN)与甲状腺癌(thyroid carcinoma，TC)细胞凋亡的关系。rn 方法:体外培养TC细胞，TERT ASODN和SODN分别作用于体外培养的TC细胞，采用透射电镜、Hoechest荧光染色、流式细胞仪等方法分析ASODN TERT对TC细胞凋亡的影响。rn 结果:透射电镜示ASODN可诱导TC出现典型的细胞凋亡形态，Hoechest荧光染色显示ASODN可使TC凋亡数目增多从48±19上升到181±28，流式细胞仪证实5 μmol/LASODN处理组、SODN处理组及空白对照组的细胞凋亡率分别为11.2％，2.1％和无凋亡(P＜0.05)。rn 结论:TERT ASODN能增强TC细胞凋亡。

摘要： 目的:研究端粒酶逆转录酶(TERT)的反义寡核苷酸(ASODN)与甲状腺癌(thyroid carcinoma，TC)细胞凋亡的关系。rn 方法:体外培养TC细胞，TERT ASODN和SODN分别作用于体外培养的TC细胞，采用透射电镜、Hoechest荧光染色、流式细胞仪等方法分析ASODN TERT对TC细胞凋亡的影响。rn 结果:透射电镜示ASODN可诱导TC出现典型的细胞凋亡形态，Hoechest荧光染色显示ASODN可使TC凋亡数目增多从48±19上升到181±28，流式细胞仪证实5 μmol/LASODN处理组、SODN处理组及空白对照组的细胞凋亡率分别为11.2％，2.1％和无凋亡(P＜0.05)。rn 结论:TERT ASODN能增强TC细胞凋亡。

摘要： 本发明提供寡核苷酸的制备方法，其包括：(1)在非极性溶剂中，使其中5’位羟基未被保护的核苷、核苷酸或寡核苷酸(a)等，与其中5’位羟基被保护的核苷、核苷酸或寡核苷酸(b)缩合，以获得含有亚磷酸三酯产物(c)的反应液的步骤；(3)使亚磷酸三酯产物(c)氧化或硫化，以获得含有其中5’位羟基被保护的寡核苷酸(d)的反应液的步骤；(4)将寡核苷酸(d)脱保护，以获得含有其中5’位羟基未被保护的寡核苷酸(e)的反应液的步骤；和(6)向含有寡核苷酸(e)的反应液中添加极性溶剂，并通过固液分离或萃取来纯化寡核苷酸(e)的步骤。

摘要： 一种由以下通式(1)至(10)中任一个表示的核苷类似物及其盐：其中R1、R2、和R3是相同或不同的基团，并且R1、R2和R3中的每一个选自氢原子、核酸合成中的官能团的保护基、磷酸基、核酸合成中由保护基保护的磷酸基和用于固相合成的活化磷酸基；并且Ar是芳香族烃基和多环芳香族烃基中的一种。

摘要： 本公开文本涉及包含有义链寡核苷酸和反义链寡核苷酸的双链寡核苷酸，包括诸如siRNA的双链寡核苷酸，并且其中所述反义链寡核苷酸包含一个或多个式(I‑A)的核苷酸类似物，所述核苷酸类似物既不是所述反义链寡核苷酸的5’‑突出端核苷酸也不是3’‑突出端核苷酸，并且其中式(I‑A)的核苷酸类似物如本公开文本所述。含有这些类似物的寡核苷酸具有优异的生物活性，例如，增加的体外稳定性和改善的体内效力，尤其是改善的脱靶概况。所改善的寡核苷酸可用于沉默(例如，减少或根除)靶基因的表达。

摘要： 本文公开了双链RNA核酸分子，其包括至少一个吡唑并三嗪核苷酸类似物且已被修饰来展现以下一者：相较于未修饰或类似修饰的dsRNA时的中靶活性增加、靶标特异性增加、核酸酶稳定性增强、脱靶活性降低和/或免疫原性降低；包含所述分子的药物组合物及其在疗法中的使用方法。

摘要： 本发明涉及使用部分立体限定的寡核苷酸子文库，鉴定反义寡核苷酸的改进的立体限定的硫代磷酸酯寡核苷酸变体的方法。这些方法允许高效鉴定具有改进特性的立体限定的变体，如增强的体外或体内活性、增强的功效、增强的比活性、降低的毒性、改变的生物分布、增强的细胞或组织摄取和/或增强的靶特异性(脱靶效应减少)。本文公开了多个平行的文库筛选方案，其中优化立体限定的硫代磷酸酯连接的核苷的多个排他性或重叠性短子区域或基序，以鉴定增强的子文库，并且随后合并来自每个选定(改进的)子文库的立体限定的核苷间键样式，以产生增强的立体限定的化合物。

摘要： 一种寡核苷酸衍生物，由通过下述一般式（一般式中，B表示腺嘌呤、鸟嘌呤、胞嘧啶或尿嘧啶，X表示硫原子或氧原子，n表示6～60的整数。）表示的重复结构单元（所述一般式中，B以及X，在各个重复结构单元中独立表示）构成，且所述一般式表示的重复结构单元中至少有一个其X是硫原子。

摘要： 本发明涉及寡核苷酸引物对、寡核苷酸组合物和包括寡核苷酸组合物的试剂盒及其检测方法，由序列为TGAYGGGCGACAATGTCC的上游引物PRRSVNSP2-F和序列为CGCAGACAAATCCAGAVG的下游引物PRRSVNSP2-R组成。本发明与现有技术相比具有以下优点：1、能够对PRRSV经典株与高致病性变异株进行鉴别诊断，解决了临床样品检测中不能区分PRRSV经典株或高致病性变异株单独感染亦或共同感染的问题；2、特异性好，针对PRRSVNSP2具有高保守区设计引物，特异性强，无假阳性出现；3、灵敏度高，该方法的敏感性可达1TCID50/mL；4、简便快速。

摘要： 本发明提供寡核苷酸的制备方法，其包括：(1)在非极性溶剂中，使其中5’位羟基未被保护的核苷、核苷酸或寡核苷酸(a)等，与其中5’位羟基被保护的核苷、核苷酸或寡核苷酸(b)缩合，以获得含有亚磷酸三酯产物(c)的反应液的步骤；(3)使亚磷酸三酯产物(c)氧化或硫化，以获得含有其中5’位羟基被保护的寡核苷酸(d)的反应液的步骤；(4)将寡核苷酸(d)脱保护，以获得含有其中5’位羟基未被保护的寡核苷酸(e)的反应液的步骤；和(6)向含有寡核苷酸(e)的反应液中添加极性溶剂，并通过固液分离或萃取来纯化寡核苷酸(e)的步骤。

摘要： 一种由以下通式(1)至(10)中任一个表示的核苷类似物及其盐：其中R1、R2、和R3是相同或不同的基团，并且R1、R2和R3中的每一个选自氢原子、核酸合成中的官能团的保护基、磷酸基、核酸合成中由保护基保护的磷酸基和用于固相合成的活化磷酸基；并且Ar是芳香族烃基和多环芳香族烃基中的一种。

摘要： 本发明提供以下述式(I)表示的立体控制寡核苷酸合成用光学活性片段及其制造方法、以及使用其的立体控制寡核苷酸的合成方法。式中，B为保护/未保护核苷碱基，R1为取代/未取代脂肪族基团，R2、R3为DMTr基或‑P(R11)(NR12)2，R11为OCH2CH2CN、SCH2CH2CN等，R12为取代/未取代的脂肪族基团或芳香族基团；X为H、烷基、O‑烷基等，Y为H、NHR13、卤素等或酰基类、醚类、硅烷基类保护羟基，或者在与X之间形成X‑Y键，n为0以上且4以下的整数。