Objective:To evaluate the effects of a booster immunization with a candidate tetanus toxoid, reduced diphtheria toxoid and acellular pertussis combined vaccine (Tdap) in a rat model after primary vaccination with diphtheria, tetanus, acellular pertussis and Sabin strain inactivated poliovirus combined vaccine (DTacP-sIPV) or diphtheria, tetanus, acellular pertussis, inactivated poliovirus and haemophilus type b combined vaccine (DTacP-IPV/Hib) for further preclinical study.Methods:Wistar rats were randomly divided into three groups and respectively immunized with a self-developed DTacP-sIPV, a marketed DTacP-IPV/Hib and normal saline at 0, 1, and 2 months of age. Serum levels of antibody against each component in each group were detected before immunization and after each dose. A booster dose of the candidate Tdap was given 10 months after primary immunization. Serum levels of antibody against each component in each group were detected before, 1 month and 6 months after the booster immunization.Results:One month after three doses of primary immunization, the geometric mean titers (GMT, Log2) of antibodies against diphtheria toxoid (DT), tetanus toxoid (TT), pertussis toxin (PT), filamentous hemagglutinin (FHA) and pertactin (PRN) in the DTacP-sIPV group were 17.41, 18.34, 18.11, 19.93 and 13.91, respectively, and the seroconversion rates of these components all reached 100%. Ten months after primary immunization, the GMTs of antibodies against DT, TT, PT, FHA and PRN decreased to 15.17, 14.26, 13.60, 14.51 and 10.39, respectively, and the seroconversion rates remained above 89%. One month after booster immunization, the GMTs of antibodies against DT, TT, PT and FHA in the DTacP-sIPV and DTacP-IPV/Hib groups were 16.49/17.26, 16.80/17.63, 16.70/17.74 and 18.48/19.26, respectively, and the seroconversion rates of these components all reached 100% with no significant difference between the two groups ( P>0.05). The GMTs of anti-PRN antibody in the DTacP-sIPV and DTacP-IPV/Hib groups were 13.07 and 11.00, and the seroconversion rates were 100% and 88%, which were higher in the DTacP-sIPV group than in the DTacP-IPV/Hib group ( P<0.05). Six months after booster immunization, the GMTs of antibodies against DT, TT, PT, FHA and PRN in the DTacP-sIPV and DTacP-IPV/Hib groups decreased to 15.74/14.87, 15.07/15.14, 14.84/15.73, 16.62/16.37 and 11.44/9.96, respectively, and the seroconversion rates remained above 88%. Conclusions:Booster vaccination with the candidate Tdap vaccine induces humoral immune response following primary immunization with DTacP-sIPV or DTacP-IPV/Hib in the Wistar rat model, while the antibody titer decreases with time.

[摘 要] 目的：检测lncRNA DNM3OS在喉鳞状细胞癌（laryngeal squamous cell carcinoma，LSCC）组织和LSCC细胞株中的表达及其临床意义，探讨其对LSCC TU177细胞体外增殖、迁移及侵袭的影响，并分析DNM3OS与EMT的关系。方法：从河北医科大学第四医院生物标本库选取2014年3月至2018年12月收治的68例LSCC患者手术切除的癌及癌旁组织标本，应用qPCR法检测DNM3OS在LSCC组织和细胞株中的表达水平。采用siRNA敲低TU177细胞中DNM3OS的表达，应用MTS、克隆形成及Transwell小室等方法分别检测敲低DNM3OS表达对TU177细胞增殖、迁移和侵袭等生物学行为的影响。应用qPCR和WB法检测转染si-DNM3OS后对EMT标志物上皮钙黏素（E-cadherin）、神经钙黏素（N-cadherin）、波形蛋白（vimentin）、扭曲蛋白（twist）、锌指转录因子2（SNAI2）mRNA和蛋白的变化。结果：LSCC组织中DNM3OS表达水平明显高于癌旁组织（P<0.01），并与患者的TNM分期、淋巴结转移及生存期有关联（P<0.05或P<0.01）。DNM3OS在LSCC细胞株（Hep-2、AMC-HN-8、TU177、TU212及TU686）中均呈现不同程度的高表达（P<0.05或P<0.01），转染si-DNM3OS后TU177细胞中DNM3OS的表达显著降低（P<0.01）。与对照组相比，DNM3OS表达敲低可抑制TU177细胞的体外增殖、迁移和侵袭能力（P<0.05或P<0.01），可上调TU177细胞中E-cadherin的表达而下调N-cadherin、vimentin、twist和SNAI2的表达（均P<0.01）。结论： DNM3OS高表达与LSCC的恶性进展有关，其可能为预测LSCC患者预后的潜在指标；DNM3OS可能通过影响EMT进程促进LSCC细胞的侵袭和转移。