目的探讨宫颈癌组织中高危型人乳头瘤病毒(high risk human papillomavirus,HR HPV)16型的致癌基因E6(HPV16-E6-DNA)病毒载量情况及其对肿瘤预后的影响。方法收集2000-06-04-2006-12-08武汉大学中南医院妇瘤科134例宫颈癌患者,均首次诊...目的探讨宫颈癌组织中高危型人乳头瘤病毒(high risk human papillomavirus,HR HPV)16型的致癌基因E6(HPV16-E6-DNA)病毒载量情况及其对肿瘤预后的影响。方法收集2000-06-04-2006-12-08武汉大学中南医院妇瘤科134例宫颈癌患者,均首次诊断为宫颈癌,行宫颈癌根治性切除术,术后经病理确诊,且随访资料完整。提取石蜡组织DNA,将HPV16-E6亚克隆到pcDNA3.1-myc-HisA载体上,利用荧光定量PCR(RT-PCR)技术检测宫颈癌组织中HPV16-E6-DNA病毒载量,并采用Spearman相关分析、Kaplan-Meier和多因素Cox比例风险回归模型分析HPV16-E6-DNA病毒载量与各临床病理特征的相关性,及其对总生存率的影响。结果 HPV16-E6成功克隆到pcDNA3.1-myc-HisA载体上。相关性分析显示,HPV16-E6-DNA病毒载量在宫颈癌组织的表达水平与宫颈癌患者年龄(P<0.001)、FIGO分期(P<0.001)和淋巴结转移(P<0.001)相关。Kaplan-Meier生存分析显示,HPV16-E6-DNA病毒拷贝数>107组5年总生存率为39.4%,105~107组为44.0%,<105组为86.3%,差异有统计学意义,P<0.001。单因素分析显示,年龄(P<0.001)、HPV16-E6-DNA病毒载量(P<0.001)、病理分级(P=0.049)、FIGO分期(P<0.001)、深肌层浸润(P=0.001)、宫旁阳性(P=0.010)和淋巴结转移(P=0.012)是影响预后的主要因素。Cox回归分析显示,淋巴结转移(P=0.002)和HPV16-E6-DNA病毒载量(P=0.010)是预后的独立危险因素。结论宫颈癌组织中,HPV16-E6-DNA病毒载量明显增加,其表达是宫颈癌根治性切除术后的总体生存率的独立预后因素,可作为宫颈癌根治性切除术后的预后预测因子之一。展开更多
目的探讨人乳头瘤病毒(HPV)E6/E7 m RNA检测在女性宫颈疾病筛查中的作用。方法选取2016年9月至2017年9月贵州医科大学附属医院妇产科门诊和住院部就诊的HPV阳性感染者287例,其中宫颈疾病患者267例[包括宫颈炎、宫颈上皮内瘤变(CIN)Ⅰ~...目的探讨人乳头瘤病毒(HPV)E6/E7 m RNA检测在女性宫颈疾病筛查中的作用。方法选取2016年9月至2017年9月贵州医科大学附属医院妇产科门诊和住院部就诊的HPV阳性感染者287例,其中宫颈疾病患者267例[包括宫颈炎、宫颈上皮内瘤变(CIN)Ⅰ~Ⅲ、宫颈癌],阳性感染者中体检无疾病表现及病理学检查无宫颈病变者20例。取患者宫颈脱落细胞用支链DNA技术(简称b-DNA技术)检测高危HPV E6/E7 m RNA,患者同时行宫颈液基薄层细胞检测和HPV分型检测,对不同年龄、HPV分型、阴道细胞学分型(简称TBS分型)及疾病分级的HPV E6/E7 m RNA的诊断价值进行效能分析。结果在不同年龄段内E6/E7阳性和阴性所占比例差异有统计学意义,P<0.05,在年龄组≤30岁组和>50~60岁组,E6/E7阳性者所占比例最大,分别为85.40%和84.40%;而HPV E6/E7m RNA拷贝数在不同年龄组内差异没有统计学意义。E6/E7结果在单一感染和混合感染,以及单一感染和多重感染间差异无统计学意义;在不同感染型别中,只有HPV 16型随病变严重程度的增加其感染率呈递增趋势(P<0.05),其余HPV型与病变严重程度无明显相关性(P>0.05)。不同疾病分组间,E6/E7阳性率的分布差异有统计学意义,表现为无宫颈病变组(0)<宫颈炎(15.60%)<CINⅠ(86.20%)<CINⅡ(93.00%)<CINⅢ(96.50%)<宫颈癌(100.00%),P<0.05;E6/E7拷贝数表达差异有统计学意义,表现为宫颈炎1773.89(936.54)<CINⅠ4997.74(10 439.75)<CINⅡ52 655.02(38 495.10)<CINⅢ72 300.12(38 298.57)<宫颈癌121 616.68(34 348.59),P<0.05。结论在宫颈疾病中检测高危型HPVE6/E7 m RNA的转录对女性宫颈癌高危人群的筛查和诊治具有重要的价值,或可作为进一步分流HPV阳性病人的方法。展开更多
RNA interference (RNAi) is triggered by the presence of a double-stranded RNA (dsRNA), and results in the silencing of homologous gene expression through the specific degradation of an mRNA containing the same sequenc...RNA interference (RNAi) is triggered by the presence of a double-stranded RNA (dsRNA), and results in the silencing of homologous gene expression through the specific degradation of an mRNA containing the same sequence. dsRNAmediated RNAi can be used in a wide variety of eucaryotes to induce the sequence-specific inhibition of gene expression.Synthetic 21-23 nucleotide (nt) small interfering RNA (siRNA) with 2 nt 3' overhangs was recently found to mediate efficient sequence-specific mRNA degradation in mammalian cells. Here, we studied the effects of synthetic siRNA duplexes targeted to SARS coronavirus structural proteins E, M, and N in a cell culture system. Among total 26 siRNA duplexes, we obtained 3 siRNA duplexes which could sequence-specifically reduce target genes expression over 80％ at the concentration of 60 nM in Vero E6 cells. The downregulation effect was in correlation with the concentrations of the siRNA duplexes in a range of 0～60 nM. Our results also showed that many inactive siRNA duplexes may be brought to life simply by unpairing the 5' end of the antisense strands. Results suggest that siRNA is capable of inhibiting SARS coronavirus genes expression and thus may be a new therapeutic strategy for treatment of SARS.展开更多
Mesoporous silica nanoparticles （MSNs） have attracted tremendous attention in recent years as drug delivery carriers due to their large surface areas, tunable sizes, facile modification and considerable biocompatibi...Mesoporous silica nanoparticles （MSNs） have attracted tremendous attention in recent years as drug delivery carriers due to their large surface areas, tunable sizes, facile modification and considerable biocompatibility. In this work, we fabricate an interesting type of MSNs which are intrinsically doped with photosensitizing molecules, chlorin e6 （Ce6）. By increasing the amount of Ce6 doped inside the silica matrix, it is found that the morphology of MSNs changes from spheres to rod-like shapes. The obtained Ce6-doped mesoporous silica nanorods （CMSNRs） are not only able to produce singlet oxygen for photodynamic therapy, but can also serve as a drug delivery platform with high drug loading capacity by utilizing their mesoporous structure. Compared to spherical nano- particles, it is found that CMSNRs with a larger aspect ratio show much faster uptake by cancer cells. With doxorubicin （DOX） employed as a model drug, the combined photodynamic and chemotherapy is carried out, achieving synergistic anti-tumor effects both in vitro and in vivo. Our study presents a new design of an MSN-based drug delivery platform, which intrinsically is fluorescent and able to serve as a photodynamic agent, promising for future imaging-guided combination therapy of cancer.展开更多
基金supported by the Grant No.2003AA208215 from the National High Technology Programs of Chinathe Grant No.30270311 from the National Natural Science Foundation of China.
文摘RNA interference (RNAi) is triggered by the presence of a double-stranded RNA (dsRNA), and results in the silencing of homologous gene expression through the specific degradation of an mRNA containing the same sequence. dsRNAmediated RNAi can be used in a wide variety of eucaryotes to induce the sequence-specific inhibition of gene expression.Synthetic 21-23 nucleotide (nt) small interfering RNA (siRNA) with 2 nt 3' overhangs was recently found to mediate efficient sequence-specific mRNA degradation in mammalian cells. Here, we studied the effects of synthetic siRNA duplexes targeted to SARS coronavirus structural proteins E, M, and N in a cell culture system. Among total 26 siRNA duplexes, we obtained 3 siRNA duplexes which could sequence-specifically reduce target genes expression over 80％ at the concentration of 60 nM in Vero E6 cells. The downregulation effect was in correlation with the concentrations of the siRNA duplexes in a range of 0～60 nM. Our results also showed that many inactive siRNA duplexes may be brought to life simply by unpairing the 5' end of the antisense strands. Results suggest that siRNA is capable of inhibiting SARS coronavirus genes expression and thus may be a new therapeutic strategy for treatment of SARS.
基金This work was partially supported by the National Basic Research Programs of China （973 Program） （Nos. 2012CB932600 and 2011CB911002）, the National Natural Science Foundation of China （Nos. 51222203 and 51132006）, Jiangsu Key Laboratory for Carbon- Based Functional Materials and Devices, a Jiangsu Natural Science Fund for Distinguished Young Scholars, and a Project Funded by the Priority Academic Program Development （PAPD） of Jiangsu Higher Education Institutions.
文摘Mesoporous silica nanoparticles （MSNs） have attracted tremendous attention in recent years as drug delivery carriers due to their large surface areas, tunable sizes, facile modification and considerable biocompatibility. In this work, we fabricate an interesting type of MSNs which are intrinsically doped with photosensitizing molecules, chlorin e6 （Ce6）. By increasing the amount of Ce6 doped inside the silica matrix, it is found that the morphology of MSNs changes from spheres to rod-like shapes. The obtained Ce6-doped mesoporous silica nanorods （CMSNRs） are not only able to produce singlet oxygen for photodynamic therapy, but can also serve as a drug delivery platform with high drug loading capacity by utilizing their mesoporous structure. Compared to spherical nano- particles, it is found that CMSNRs with a larger aspect ratio show much faster uptake by cancer cells. With doxorubicin （DOX） employed as a model drug, the combined photodynamic and chemotherapy is carried out, achieving synergistic anti-tumor effects both in vitro and in vivo. Our study presents a new design of an MSN-based drug delivery platform, which intrinsically is fluorescent and able to serve as a photodynamic agent, promising for future imaging-guided combination therapy of cancer.
