Inactivated influenza vaccine formulated with single-stranded RNA-based adjuvant confers mucosal immunity and cross-protection against influenza virus infection

Influenza vaccination is considered the most valuable means to prevent and control seasonal influenza infections, which causes various clinical symptoms, ranging from mild cough and fever to even death. Among various influenza vaccine types, the inactivated subunit type is known to provide improved safety with reduced reactogenicity. However, there are some drawbacks associated with inactivated subunit type vaccines, with the main ones being its low immunogenicity and the induction of Th2-biased immune responses. In this study, we investigated the role of a single-stranded RNA (ssRNA) derived from the intergenic region in the internal ribosome entry site of the Cricket paralysis virus as an adjuvant rather than the universal vaccine for a seasonal inactivated subunit influenza vaccine. The ssRNA adjuvant stimulated not only well-balanced cellular (indicated by IgG2a, IFN-γ, IL-2, and TNF-α) and humoral (indicated by IgG1 and IL-4) immune responses but also a mucosal immune response (indicated by IgA), a key protector against respiratory virus infections. It also increases the HI titer, the surrogate marker of influenza vaccine efficacy. Furthermore, ssRNA adjuvant confers cross-protective immune responses against heterologous influenza virus infection while promoting enhanced viral clearance. Moreover, ssRNA adjuvant increases the number of memory CD4⁺ and CD8⁺ T cells, which can be expected to induce long-term immune responses. Therefore, this ssRNA-adjuvanted seasonal inactivated subunit influenza vaccine might be the best influenza vaccine generating robust humoral and cellular immune responses and conferring cross-protective and long-term immunity.     

WT1基因与急性髓系白血病的预后及主动特异性免疫治疗的研究进展

WT1基因位于人类染色体11p13，在80%的急性髓系白血病（acute myeloid leukemia，AML）患者中高表达，是AML预后不良的分子标志，可作为AML预后评估和微小残留病变（minimal residual disease，MRD）监测的有效指标。由于WT1在AML中均有异常高表达，故认为是一种AML抗原，可作为特异性免疫治疗的新靶点。一些小规模的临床试验证实以WT1为靶点的免疫治疗是有效的、安全的，这些免疫治疗可作为那些有高危复发风险及初始标准化疗失败的AML患者的辅助治疗。本文就近几年WT1与AML的预后及有关以WT1为靶点的主动特异性免疫治疗的研究进展予以综述。     

极速实时荧光聚合酶链反应与常规方法对新型布尼亚病毒检测的评价

目的探讨极速实时荧光聚合酶链反应(polymerase chain reaction,PCR)、实时荧光PCR、酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)和胶体金免疫层析法(gold immunochromatography assay,GICA)4种方法检测新型布尼亚病毒的特异度和灵敏度,为发热伴血小板减少综合征的早期诊断提供依据。方法采集2017年6月1日至9月30日山东大学附属济南市传染病医院86例临床诊断为发热伴血小板减少综合征患者的血清样本,分别应用极速实时荧光PCR、实时荧光PCR、ELISA和GICA 4种方法进行检测。统计学分析采用χ^2检验。结果86份患者血清标本中,极速实时荧光PCR、实时荧光PCR、IgM-ELISA、IgG-ELISA、IgM-GICA、IgG-GICA的新型布尼亚病毒阳性分别为82份(95.34%)、79份(91.86%)、41份(47.67%)、8份(9.3%)、19份(22.09%)和3份(3.49%)。极速实时荧光PCR特异度为100%,灵敏度达到1×103拷贝/mL,3次重复扩增试验显示其Ct值变异系数均<2%。在发热伴血小板减少综合征进展的1期、2期、3期病程中,极速实时荧光PCR的阳性检出率为41份(97.62%)、34份(94.44%)、7份(87.50%),实时荧光PCR的阳性检出率为39份(92.86%)、33份(91.67%)、7份(87.50%),在1期和2期两个病程,极速实时荧光PCR阳性检出率略高;IgM-ELISA阳性检出率从1期(28.57%)到3期(87.50%)显著增高,2期、3期与1期相比,差异均有统计学意义(χ^2=8.347、7.561,均P<0.01);IgM-GICA的阳性检出率从1期(14.29%)到2期(33.33%)也有增高,差异有统计学意义(χ^2=3.962,P<0.05),但与其他方法相比,其检出率偏低。1期,实时荧光PCR阳性检出率显著高于ELISA(IgM和IgG)和GICA(IgM和IgG),差异均有统计学意义(χ^2=33.740、55.080、49.010、64.340,均P<0.01)。2期,实时荧光PCR的阳性检出率高于ELISA(IgM和IgG)和GICA(IgM和IgG),差异均有统计学意义(χ^2=7.700、46.720、23.700、50.630,均P<0.01)。3期,极速实时荧光PCR、实时荧光PCR和IgM-ELISA表现出同样高的阳性检出率,远高于IgG-ELISA和GICA(IgM和IgG)。实时荧光PCR阳性检出率和IgG-ELISA、IgM-GICA、IgG-GICA之间差异均有统计学意义(均χ^2=6.250,P<0.05)。结论极速实时荧光PCR在新型布尼亚病毒的早期检测中有更高的灵敏度和特异度,且重复性好、稳定度高,与传统实时荧光PCR相比大大缩短了扩增时间,对发热伴血小板减少综合征的早期快速诊断具有重要价值。     

Translocation between chromosome 5q35 and chromosome 11q13 – an unusual cytogenetic finding in a primary refractory acute myeloid leukemia

Cytogenetic abnormalities are observed in approximately two‐thirds of patients with acute myeloid leukemia (AML). Chromosome rearrangements are associated with specific subtypes of AML and associated prognosis. We report a patient with AML, M2, who was primarily refractory to standard induction chemotherapy with idarubicin and cytarabine. Flow cytometry of a bone marrow aspirate showed aberrant expression of B‐cell markers including CD19. Cytogenetic studies disclosed a translocation between 5q35 and 11q13. Fluorescence in situ hybridization analyses demonstrated that neither the NSD1 nor MLL genes were involved in this case. Further study is required to define conclusively the genes involved and their contribution to pathogenesis in this case.     

中国优秀马拉松运动员ACE基因I/D多态性频率分布特征

通过分析中国马拉松运动员ACE基因I/D多态频率分布特征,探讨其作为杰出耐力基因标记的可行性。选择我国马拉松健将、国际健将级运动员26名作为马拉松运动员组,汉族学生216名作为对照组。对两组受试者进行ACE基因I/D多态性测定。结果显示:我国马拉松运动员组的等位基因频率和基因型频率与对照组比较无显著差异,其中15名国际健将中无一DD型纯合子,提示我国优秀马拉松运动员的纯合子DD型频率低下是其ACE基因多态频率分布的主要特征。     

人间充质干细胞免疫原性研究

目的探讨人间充质干细胞(hMSCs)特性及免疫原性,为进一步研究hMSCs移植特性提供实验依据。方法免疫组化方法鉴定hMSCs表面HLA-ABC、HLA-DR、CD80、CD86分子;流式细胞术检测其含量;RT-PCR检测HLA-ABC、HLA-DRmRNA基因片断;外周血淋巴细胞杀伤试验及 CCK-8比色法规察淋巴细胞增殖反应;将表达绿色荧光蛋白(GFP)的DNA复合物导入hMSCs进行大鼠脑内移植,观察hMSCs在脑内的存活情况。结果 hMSCs表面少量表达HLA-ABC分子,不表达 HLA-DR、CD80、CD86分子;有少量HLA-ABC mRNA基因片断存在,未发现HLA-DR mRNA基因片断;外周血淋巴细胞杀伤试验没有发现淋巴细胞增殖反应;大鼠脑内移植hMSCs一个月后仍可见有细胞存活。结论 hMSCs具有较弱的免疫原性。     

Histological and biological developmental characterization of the human cerebellar cortex

The aim of this study was to investigate the histological and biological features of the human cerebellar cortex development and differentiation. We analyzed 52 brains of fetal and infant death victims, aged from 17 gestational weeks to 12th postnatal month. In particular, in the cerebellar cortex at different ages we evaluated, besides the structural aspects, the expression of several biomarkers implicated in proliferative processes (c-fos, PCNA and apoptosis). We observed morphological patterns progressively evolving every month, from the indefinite structure of the second gestational trimester to the four-layered structure (external granular layer, molecular layer, Purkinje cell layer, internal granular layer) of the late fetal cortex and subsequently to the three-layered postnatal definitive morphology, due to involution of the external granular layer. The evaluation of the biological features of the cerebellar cortex showed high proliferative activity mainly confined to the transient external granular layer in prenatal life, and high apoptotic index after birth. Thus, the histological examination, better with the support of biomarker investigations, allows with accuracy to describe the dynamic sequence of steps that occur in human cerebellar cortex development and to establish in each case the age, namely the pre- or postnatal month of life. Consequently, we can diagnose delayed or altered processes of differentiation during the development of the human cerebellar cortex.     

移植人羊膜细胞对大鼠创伤性脑损伤的实验研究

目的 探究大鼠TBI后脑内移植人羊膜细胞（HACs）对大鼠运动功能的影响。方法 HACs经分离、Hoechst33342标记后重悬调整细胞浓度为10^5/μl；采用改进的Feeney自由落体法打击大鼠脑皮层后肢运动区域，损伤后24h经微量注射器和立体定向仪将Hoechst33342标记的HACs 10μl分别移植于挫伤灶中心和挫伤灶边缘；在TBI后的28d内采用钉板平衡木行走测试大鼠运动功能变化，运动功能检测结束后取出脑组织行组织学检测。结果 治疗组滑落脚步数明显少于对照组（P〈0．05）；移植的HACs呈蓝色荧光；部分移植HACs可见MAP-2阳性表达。结论 移植HACs使大鼠TBI后运动功能明显改善。     

增强子Ⅰ对乙型肝炎病毒DNA疫苗免疫应答的影响

目的 研究乙型肝炎病毒（HBV）自身增强子I（enhancerI，ENHI）对HBV DNA疫苗免疫应答的影响。方法 采用PCR法以HBVadr亚型全基因DNA序列为模板分别扩增表面抗原（HBsAg）和HBsA-ENHI基因片段，重组到载体VR1012中，构建两种HBV DNA疫苗，转染CADS-7细胞及HepG2细胞并免疫BALB／c小鼠。采用蛋白印迹、ELISA、ELISPOT等方法检测其在COS-7和HepG2细胞内的表达及小鼠的体液及细胞免疫应答效果。结果 转染的HepG2和COS-7细胞均表达HBsAg;连接ENHI的HBV DNA疫苗转染HepG2细胞后HBsAg表达量明显升高，两种疫苗转染COS-7细胞表达HBsAg无明显差异；免疫小鼠后第2周产生HBsAb及HBsAg特异性细胞毒T淋巴细胞（CTL），两种疫苗免疫产生的HBsAb及HBsAg特异性CTL无明显差异。结论ENHI可使HBV DNA疫苗转染HepG2细胞表达HBsAg明显增加，对转染COS-7细胞表达HBsAg及接种BALB／C小鼠引起的免疫应答无明显影响。