目的探讨HTR3B基因多态性与头颈部恶性肿瘤患者化疗后出现恶心呕吐的关系。方法采用方便抽样的方法,选择行TPF方案化疗的头颈部恶性肿瘤患者212例。从NCBI数据库或既往相关文献中筛选有临床意义的HTR3B基因rs3758987、rs1176744、rs1279...目的探讨HTR3B基因多态性与头颈部恶性肿瘤患者化疗后出现恶心呕吐的关系。方法采用方便抽样的方法,选择行TPF方案化疗的头颈部恶性肿瘤患者212例。从NCBI数据库或既往相关文献中筛选有临床意义的HTR3B基因rs3758987、rs1176744、rs12795805、rs2276305位点,化疗前采集患者外周静脉血3 m L,检测各位点的基因型。分析各位点基因型与化疗后出现恶心呕吐的关系。结果 212例患者化疗后出现恶心呕吐113例,出现恶心呕吐与rs3758987位点有关(P<0.05),与rs1176744、rs12795805、rs2276305位点无关(P均>0.05)。HTR3B基因rs3758987位点TT基因型者化疗后恶心呕吐的发生率明显低于CC+CT基因型者(P<0.05)。结论HTR3B基因rs3758987位点与头颈部恶性肿瘤患者化疗后出现恶心呕吐有关,其CC、CT基因型者化疗后出现恶心呕吐的风险明显高于TT基因型者。展开更多
从口蹄疫病毒(foot-and-m ou th d isease v irus,FM DV)细胞培养物中提取总RNA,经一步法RT-PCR获得了长约230 bp的3B基因片段,将PCR产物连接到pM D-18T载体上并测序,用B amHⅠ与H indⅢ双酶切后,将3B基因融合到pGEX-KG载体上形成表达质...从口蹄疫病毒(foot-and-m ou th d isease v irus,FM DV)细胞培养物中提取总RNA,经一步法RT-PCR获得了长约230 bp的3B基因片段,将PCR产物连接到pM D-18T载体上并测序,用B amHⅠ与H indⅢ双酶切后,将3B基因融合到pGEX-KG载体上形成表达质粒pGEX-KG-3B,转化到BL 21(DE 3)中在27℃诱导表达,将表达产物进行SDS-PAGE和蛋白质斑点EL ISA。结果表明3B基因主要以可溶形式高效表达,表达产物具有免疫原性。以纯化的表达产物为抗原建立了间接酶联免疫吸附试验(I-EL ISA)。方阵滴定其最佳包被浓度是6.1μg/孔,最佳血清稀释倍数为1∶80,通过测定36份FM DV阴性血清,确定了该方法的阳性判定标准。结果表明,该方法特异性强、重复性好;用3B-EL ISA方法与美国联合生物医学公司(U B I)生产的合成肽检测试剂盒U B I○RFM DV N S-EL ISA对比检测44份血清样品,符合率为93.1%。证明3B-EL ISA方法可用于口蹄疫的鉴别诊断。展开更多
1b and 2a HCV subtypes we re mainly reported in ChinaHerein ,a 3b subtype strain of HCV was fo und with gene chip methodFirstly, the 5’ NCR(non-coding region)was s elected as target sequence,and 171 related seque...1b and 2a HCV subtypes we re mainly reported in ChinaHerein ,a 3b subtype strain of HCV was fo und with gene chip methodFirstly, the 5’ NCR(non-coding region)was s elected as target sequence,and 171 related sequences in GeneBank wer e alignedAccording to the alignme nt results,eleven DNA probes were acquired and oligonucleotide array s were constructed for distinguish ing the HCV main genotypes and sub types,which did not contain specif ic probes of genotypes 4 and 5,but genotype 1, subtypes 1a/1c and 1b, genotype 2,subtypes 2a/2c and 2b,g enotype 3,subtypes 3a and 3b and g enotype 6,subtype 6aSecondly,foll owing reverse transcription of HCV RNA,nested-PCR was applied to harv est the 260 bp target DNA fragment approximatelyAfter specific ampli fication of target DNA and array h ybridization,the solid phase alkal ine phosphatase reaction indicated visually the results that the DNA fragment contains specific sequenc e of genotype 3 and subtype 3bBy sequencing the target DNA fragment ,and analyzing the sequence,it con firmed that HCV containing in the p ositive blood of the patient is gen otype 3 and subtype展开更多
文摘目的探讨HTR3B基因多态性与头颈部恶性肿瘤患者化疗后出现恶心呕吐的关系。方法采用方便抽样的方法,选择行TPF方案化疗的头颈部恶性肿瘤患者212例。从NCBI数据库或既往相关文献中筛选有临床意义的HTR3B基因rs3758987、rs1176744、rs12795805、rs2276305位点,化疗前采集患者外周静脉血3 m L,检测各位点的基因型。分析各位点基因型与化疗后出现恶心呕吐的关系。结果 212例患者化疗后出现恶心呕吐113例,出现恶心呕吐与rs3758987位点有关(P<0.05),与rs1176744、rs12795805、rs2276305位点无关(P均>0.05)。HTR3B基因rs3758987位点TT基因型者化疗后恶心呕吐的发生率明显低于CC+CT基因型者(P<0.05)。结论HTR3B基因rs3758987位点与头颈部恶性肿瘤患者化疗后出现恶心呕吐有关,其CC、CT基因型者化疗后出现恶心呕吐的风险明显高于TT基因型者。
文摘从口蹄疫病毒(foot-and-m ou th d isease v irus,FM DV)细胞培养物中提取总RNA,经一步法RT-PCR获得了长约230 bp的3B基因片段,将PCR产物连接到pM D-18T载体上并测序,用B amHⅠ与H indⅢ双酶切后,将3B基因融合到pGEX-KG载体上形成表达质粒pGEX-KG-3B,转化到BL 21(DE 3)中在27℃诱导表达,将表达产物进行SDS-PAGE和蛋白质斑点EL ISA。结果表明3B基因主要以可溶形式高效表达,表达产物具有免疫原性。以纯化的表达产物为抗原建立了间接酶联免疫吸附试验(I-EL ISA)。方阵滴定其最佳包被浓度是6.1μg/孔,最佳血清稀释倍数为1∶80,通过测定36份FM DV阴性血清,确定了该方法的阳性判定标准。结果表明,该方法特异性强、重复性好;用3B-EL ISA方法与美国联合生物医学公司(U B I)生产的合成肽检测试剂盒U B I○RFM DV N S-EL ISA对比检测44份血清样品,符合率为93.1%。证明3B-EL ISA方法可用于口蹄疫的鉴别诊断。
文摘1b and 2a HCV subtypes we re mainly reported in ChinaHerein ,a 3b subtype strain of HCV was fo und with gene chip methodFirstly, the 5’ NCR(non-coding region)was s elected as target sequence,and 171 related sequences in GeneBank wer e alignedAccording to the alignme nt results,eleven DNA probes were acquired and oligonucleotide array s were constructed for distinguish ing the HCV main genotypes and sub types,which did not contain specif ic probes of genotypes 4 and 5,but genotype 1, subtypes 1a/1c and 1b, genotype 2,subtypes 2a/2c and 2b,g enotype 3,subtypes 3a and 3b and g enotype 6,subtype 6aSecondly,foll owing reverse transcription of HCV RNA,nested-PCR was applied to harv est the 260 bp target DNA fragment approximatelyAfter specific ampli fication of target DNA and array h ybridization,the solid phase alkal ine phosphatase reaction indicated visually the results that the DNA fragment contains specific sequenc e of genotype 3 and subtype 3bBy sequencing the target DNA fragment ,and analyzing the sequence,it con firmed that HCV containing in the p ositive blood of the patient is gen otype 3 and subtype
