Evi1和MDS1基因

Evil基因在小鼠逆转录病毒诱发的急性髓系白血病(AML)中,是病毒常见的插入位点,在AML发病中起重要作用。MDS基因位于Evil上游,通过选择性剪接,可形成MDS1-Evil基因。本文就Evi-1和MDS1基因的研究近况作一综述。     

FoxO1 基因对Jurkat 细胞FoxO1-KLF2-S1P1 通路的调节作用

目的:通过构建FoxO1 表达和干扰慢病毒载体,建立细胞内FoxO1-KLF2-S1P1 信号通路调控研究模型,观察FoxO1 过表达、干扰表达在Jurkat 细胞内对其下游分子表达及功能的影响。方法:构建FoxO1 表达和干扰表达慢病毒载体,分别感染Jurkat 细胞,采用荧光定量PCR、Western blot 和流式细胞术检测S1P1、CD62L、CCR7、CD69 mRNA 水平和蛋白分子的表达。结果:FoxO1 过表达组于感染后120 h FoxO1、KLF2、S1P1 和CD62L mRNA 水平显著增高(P<0.05),FoxO1、FoxO1-p 和KLF2 胞浆蛋白水平增高,S1P1+细胞和CD62L+ 细胞比率增高(P<0.05),CCR7+ 细胞和CD69+ 细胞未见显著改变(P>0.05)。FoxO1 干扰组于转染后120 h FoxO1、KLF2、S1P1 和CD62L mRNA 水平降低(P<0.05),FoxO1、FoxO1-p 和KLF2 胞浆蛋白水平低于对照组,S1P1+细胞百分比增多(P<0.05) ,但S1P1+细胞和CD62L+细胞在72 h 时减少(P<0.05)。结论:FoxO1 表达和干扰慢病毒载体转染Jurkat 细胞并调节KLF2、S1P1 和CD62L 等分子的表达,为开展细胞内FoxO1-KLF2-S1P1 信号通路调控和细胞相关功能的研究打下了基础。     

韩国人CYP450 1A1、CYP450 2E1和GSTM1、GSTT1、GSTP1基因座遗传多态性分析

目的 调查代谢相关的CYP4501A1、CYP4502E1和GSTM1、GSIT1、GSTP1基因座在韩国人群中的遗传多态性分布状况。方法 采用多重聚合酶链式反应、聚合酶链式反应-限制性片段长度多态性技术，分析300名韩国健康大学生的CYP1A1基因3′端限制性内切酶Msp Ⅰ位点、CYP2E1基因5′端转录调节区Pst Ⅰ位点和GSTM1、GSTT1缺失与存在、GSTP1基因第5外显子BsmA Ⅰ位点的基因型，计算基因型和基因频率。结果 CYP1A1基因型频率为ml／ml型39．7％、ml／m2型49．7％、m2／m2型10．7％，基因频率为ml 0．645、m2 0．355。CYP2E1基因型频率为cl／cl型66．7％、cl／c2型30％、c2／c2型3．3％，基因频率为C1 0．818、C2 0．182。GSTM1基因缺失型频率为53．3％。GSTT1基因缺失型频率为54．7％。GSTP1基因型频率为Ile／Ile型62％、Ile／Val型34．3％、VaL／Val型3．7％，基因频率为Ile 0．792、Val 0．208。基因分布符合Hardy-Weirtberg平衡定律。结论 韩国人CYP1A1、CYP2E1、GSTM1、GSTT1基因分布与我国人群较为相近，半数以上人缺乏GSTM1和GSTT1基因，纯合缺失型频率超过印度人的3倍。     

先兆子痫患者HLA-DQA1、-DQB1、-DPA1基因多态性

目的:探讨人类白细胞抗原HLA-DQ-A1、DQB1、DPA1基因多态性与先兆子痫发病的关系。方法:采用序列特异性引物技术(PCRSSP)对46例先兆子痫患者和105例正常孕妇及其新生儿进行HLA-DQ-DPA1等位基因分型。结果:所有标本共检出11种HLADQA1基因表型、16种HLADQB1基因表型、6种HLADPA1基因表型。先兆子痫患者HLA-DQ-B10301基因频率高于正常孕妇,差异有显著性(Pc=0.032,RR=2.43,AR=0.30),其余各基因表型频率两组比较差异均无显著性。结论:HLADQB10301基因可能是一种先兆子痫发病的易感基因。     

AKR1C1在恶性肿瘤中的研究进展

人20α-羟基类固醇脱氢酶(aldo-keto reductase family 1member C1,AKR1C1)是醛酮还原酶(aldo-keto reductase,AKRs)超家族成员之一。AKR1C1的功能:(1)调节雄激素、雌激素和孕激素的代谢,参与多种生理功能,与类固醇激素依赖型恶性肿瘤密切相关;(2) AKR1C1的过表达可降低化疗的敏感性,增加化疗耐药性,是导致化疗失败的主要原因。近年有研究已证实AKR1C1对肿瘤的侵袭、转移、化疗药物耐药有着重要的作用。因此,分析AKR1C1对肿瘤发生、发展的分子机制及靶向治疗有重要意义。该文针对AKR1C1在恶性肿瘤中的研究进展作一综述。     

注射甲型H1N1流感疫苗引起亚急性甲状腺炎1例报告

亚急性甲状腺炎（subacute thyroiditis,SAT）简称亚甲炎,由De Quervain于1940年首先描述,是临床上较为常见的甲状腺疾病.亚急性甲状腺炎的发病机制虽然尚未明了,但一般认为与感染有关,最常见的病因是由于各种病毒感染所致,病原菌包括流感病毒、柯萨奇病毒、腺病毒、腮腺炎病毒等.近年来认为该病与病毒感染后引起的免疫功能紊乱有关,属免疫反应性疾病.由注射疫苗引发的亚甲炎国内外罕见报导,现将一例注射甲型H1N1流感疫苗引发的亚甲炎报告如下.     

C1q、C1r和C1s的快速分离

建立了一种同时快速分离纯化C1q及酶原形式C1r和C1s的方法。用IgG Sepharose 4B亲和层析将C1q与C1r2 s2 分离 ,接着用DEAE Sephacel离子交换层析将C1r和C1s分离 ,用C1qMcAb Sepharose 4B亲和层析将C1q进一步纯化。在分离Clr和C1s的整个过程中加入蛋白水解抑制剂PMSF和NPGB ,并控制温度在 4℃、pH 6 1、无二价阳离子的条件下 ,得到高度纯化的C1q、C1r和C1s。     

流感病毒H1N1感染后宿主细胞MTH1基因表达的研究

目的观察H1N1型流感病毒诱导的MDCK细胞抗氧化相关修复基因MTH1表达量的变化。方法 MDCK细胞培养后,H1N1型流感病毒感染0、1、3、6、12、24、48h,在各时间点经RT-PCR反应检测细胞内MTH1基因的相对表达量。结果甲型H1N1流感病毒感染后0、6、12h,MTH1基因的表达量与正常对照组差异无统计学意义;感染后1、3hMTH1基因的表达量显著高于正常对照组;感染后24、48h表达量显著低于正常对照组。结论 H1N1流感病毒感染细胞的MTH1基因表达的增加,可能增加细胞对DNA氧化损伤的修复能力,在流感发生和防御机制中起到作用。     

1例接种甲型H1N1流感疫苗后心因性反应案例分析

1病例基本情况 患者于2009年11月25日15：40接种甲流疫苗1h后出现头晕、恶心、腹痛、右膝关节痛等症状,经现场临床医生查体,仅有下腹部轻压痛,右膝关节皮肤稍触及疼痛明显的阳性体征,经双侧内关、足三里穴针炙后,欲吐止,腹痛及右膝关节痛缓解,     

Protective immunity against influenza H5N1 virus challenge in mice by intranasal co-administration of baculovirus surface-displayed HA and recombinant CTB as an adjuvant

The increasing number of recent outbreaks of HPAI H5N1 in birds and humans brings out an urgent need to develop potent H5N1 vaccine regimens. Here we present a study on the intranasal vaccination of recombinant baculovirus surface-displayed hemagglutinin (BacHA) or inactivated whole H5N1 viral (IWV) vaccine with a recombinant cholera toxin B subunit (rCTB) as a mucosal adjuvant in a BALB/c mouse model. Two groups of mice were vaccinated with different doses (HA titer of log 2⁴ or log 2⁸) of either HA surface-displayed baculovirus or inactivated whole viral vaccine virus adjuvanted with different doses (2 μg or 10 μg) of rCTB. The vaccinations were repeated after 28 days. HA specific serum IgG and mucosal IgA antibodies were quantified by indirect ELISA, and serum neutralizing antibody titer were estimated by hemagglutination inhibition (HI) assay and virus neutralization titer assay. Functional protective efficacy of the vaccine was assessed by host challenge against HPAI H5N1 strains. The results revealed that mice co-administered with log 2⁸ HA titer of BacHA vaccine and adjuvanted with 10 μg of rCTB had a significantly enhanced serum IgG and mucosal IgA immune response and serum microneutralization titer compared with mice administered with unadjuvanted log 2⁴ or log 2⁸ HA titer of BacHA alone. Also vaccination with 10 μg of rCTB and log 2⁸ HA titer of BacHA elicited higher HA specific serum and mucosal antibody levels and serum HI titer than vaccination with log 2⁸ HA titer of inactivated H5N1 virus adjuvanted with the same dose of rCTB. The host challenge study also showed that 10 μg rCTB combined with log 2⁸ HA titer of BacHA provided 100% protection against 10MLD₅₀ of homologous and heterologous H5N1 strains. The study shows that the combination of rH5 HA expressed on baculovirus surface and rCTB mucosal adjuvant form an effective mucosal vaccine against H5N1 infection. This baculovirus surface-displayed vaccine is more efficacious than inactivated H5N1 influenza vaccine when administered by intranasal route and has no biosafety concerns associated with isolation, purification and production of the latter vaccine.     

Immunization with a low dose of hemagglutinin-encoding plasmid protects against 2009 H1N1 pandemic influenza virus in mice

A vaccine against the novel pandemic influenza virus (2009 H1N1) is available, but several problems in preparation of vaccines against the new emerging influenza viruses need to be overcome. DNA vaccines represent a novel and powerful alternative to conventional vaccine approaches. To evaluate the ability of a DNA vaccine encoding the hemagglutinin (HA) of 2009 H1N1 to generate humoral responses and protective immunity, BALB/c mice were immunized with various doses of 2009 H1N1 HA-encoding plasmid and anti-HA total IgG, hemagglutination inhibition antibodies and neutralizing antibodies were assayed. The total IgG titers against HA correlated positively with the doses of DNA vaccine, but immunization with either a low dose (10 μg) or a higher dose (25-200 μg) of HA plasmid resulted in similar titers of hemagglutination inhibition and neutralizing antibodies, following a single booster. Further, 10 μg plasmid conferred effective protection against lethal virus challenge. These results suggested that the DNA vaccine encoding the HA of 2009 H1N1 virus is highly effective for inducing neutralizing antibodies and protective immunity. DNA vaccines are a promising new strategy for the rapid development of efficient vaccines to control new emerging pandemic influenza viruses.     

Enhanced growth of seed viruses for H5N1 influenza vaccines

Seed viruses used to produce inactivated H5N1 influenza vaccines are recombinant viruses with modified avirulent-type hemagglutinin (HA) and intact neuraminidase (NA) genes, both derived from an H5N1 isolate, and all remaining genes from the PR8 strain, which grows well in eggs. However, some reassortants grow suboptimally in eggs, imposing obstacles to timely, cost-efficient vaccine production. Here, we demonstrate that our PR8 strain supports better in ovo growth than the PR8 strain used for the WHO-recommended seed virus, NIBRG-14. Moreover, inclusion of an alternative NA protein further enhanced viral growth in eggs. These findings suggest that our H5N1 vaccine candidates would increase the availability of H5N1 vaccine doses at the onset of a new pandemic.     

2009 H1N1 influenza and experience in three critical care units

Aim: We describe futures of ICU admission, demographic characteristics, treatment and outcome for critically ill patients with laboratory-confirmed and suspected infection with the H1N1 virus admitted to the three different critical care departments in Turkey.Methods: Retrospective study of critically ill patients with 2009 influenza A(H1N1) at ICU. Demographic data, symptoms, comorbid conditions, and clinical outcomes were collected using a case report form.Results: Critical illness occurred in 61 patients admitted to an ICU with confirmed (n=45) or probable and suspected 2009 influenza A(H1N1). Patients were young (mean, 41.5 years), were female (54%). Fifty-six patients, required mechanical ventilation (14 invasive, 27 noninvasive, 15 both) during the course of ICU. On admission, mean APACHE II score was 18.7±6.3 and median PaO₂/FIO₂ was 127.9±70.4. 31 patients (50.8%) was die. There were no significant differences in baseline PaO₂/FIO₂ and ventilation strategies between survivors and nonsurvivors. Patients who survived were more likely to have NIMV use at the time of admission to the ICU.Conclusion: Critical illness from 2009 influenza A(H1N1) in ICU predominantly affects young patients with little major comorbidity and had a high case-fatality rate. NIMV could be used in 2009 influenza A (H1N1) infection-related hypoxemic respiratory failure.     

The safety and immunogenicity of a cell-derived adjuvanted H5N1 vaccine – A phase I randomized clinical trial

BackgroundDevelopment of an efficacious egg-free mock-up H5N1 vaccine is key to our preparedness against pandemic avian flu.MethodsThis is a single-center, randomized, observer-blinded phase I clinical trial evaluating the safety and immunogenicity of an alum-adjuvanted Madin–Darby canine kidney (MDCK)-derived inactivated whole-virion H5N1 influenza vaccine in healthy adults. Hemagglutination inhibition (HAI) and neutralizing antibody titers were measured using horse and turkey red blood cells (RBCs).ResultsThirty-six adult subjects were randomized to receive two doses of 0.5 mL of the MDCK-derived H5N1 alum-adjuvanted vaccine containing 7.5, 15, or 30 μg of hemagglutinin (HA) 21 days apart. The candidate vaccine was well tolerated and safe across the three dosing groups. The most frequent adverse event was injection site pain (46.5%). Both HAI and neutralizing antibody titers increased after each vaccination in all three dosing groups. The best HAI responses, namely a seroconversion rate of 91.7% and a geometric mean ratio of 9.51 were achieved with the HA dose of 30 μg assayed using horse RBCs at day 42. HAI titers against H5N1 avian influenza virus was significantly higher when measured using horse RBCs compared with turkey RBCs.ConclusionsThis Phase I trial showed the MDCK-derived H5N1 candidate vaccine is safe and immunogenic. The source of RBCs has a significant impact on the measurement of HAI titers (ClinicalTrials.gov number: NCT01675284.).     

H1亚型流感病毒HA蛋白单克隆抗体的制备及部分特性鉴定

目的:制备针对H1亚型流感病毒HA蛋白的单克隆抗体(mAb),并分析其反应特性。方法:分别以2009年甲型H1N1、季节性A1流感病毒裂解疫苗为免疫原,常规法免疫、融合、克隆化,获得各抗原特异性mAb。应用ELISA、HI试验和Western blot等技术研究mAb的反应性和特异性。结果:获得稳定分泌抗H1亚型流感病毒HA蛋白的杂交瘤细胞97株。其中株特异性mAb39株,29株具有HI活性;亚型特异性mAb7株,5株具有HI活性;2009年流行株与季节性A1、A3流行株共同抗原的mAb16株,9株具有HI活性;针对流感病毒共同抗原mAb35株,22株具有HI活性。结论:两种疫苗均具有较好的免疫原性和免疫保护活性,这些mAb的获得为流感病毒株特异、亚型特异性诊断试剂盒及流感病毒通用诊断试剂盒的制备提供了实验资料,为进一步研究H1N1流感病毒HA的抗原表位奠定了基础。     

Genetic analysis of HA gene of pandemic H1N1 2009 influenza viruses circulating in India

The H1N1 2009 influenza pandemic took the health care workers by surprise in spite of warning about influenza pandemic. Influenza A virus has the ability to overcome immunity from previous infections through the acquisition of genetic changes by shift or drift. Thus, understanding the evolution of the viruses in human is important for the surveillance and the selection of vaccine strains. A total of 23 pandemic A/H1N1 2009 viral HA gene sequences were downloaded from NCBI submitted during March and May 2010 by NIV and were analysed. Along with that the vaccine strain A/California/07/2009 was also downloaded from NCBI. All the sequences were used to analyse the evolution of the haemagglutinin (HA) by phylogenetic analysis. The HA gene could be divided into four groups with shift from 1 to lV revealing that the HA genes of the influenza A viruses evolved in a sequential way, in comparison to vaccine strain A/California/07/2009. Amino acid sequence analysis of the HA genes of the A/H1N1 2009 isolates, revealed mutations at positions 100, 220 and additional mutations in different positions 114, 171, 179, 190, 208, 219, 222, 239, 240, 247, 251, 260 and 285 .The mutations identified showed the adaptation of the new virus to the host that could lead to genetic changes inherent to the virus resulting in a reassortant which could be catastrophic, hence continuous monitoring of strains is mandatory.     

Molecular characterization of the influenza A(H1N1)pdm09 isolates collected in the 2015-2016 season and comparison of HA mutations detected in Turkey since 2009

Influenza A(H1N1)pdm09 pandemic virus causing the 2009 global outbreak moved into the post-pandemic period, but its variants continued to be the prevailing subtype in the 2015-2016 influenza season in Europe and Asia. To determine the molecular characteristics of influenza A(H1N1)pdm09 isolates circulating during the 2015-2016 season in Turkey, we identified mutations in the hemagglutinin (HA) genes and investigated the presence of H275Y alteration in the neuraminidase genes in the randomly selected isolates. The comparison of the HA nucleotide sequences revealed a very high homology (>99.5%) among the studied influenza A(H1N1)pdm09 isolates, while a relatively low homology (96.6%-97.2%), was observed between Turkish isolates and the A/California/07/2009 vaccine virus. Overall 14 common mutations were detected in HA sequences of all 2015-2016 influenza A(H1N1)pdm09 isolates with respect to the A/California/07/2009 virus, four of which located in three different antigenic sites. Eleven rare mutations in 12 HA sequences were also detected. Phylogenetic analysis revealed that all characterized influenza A(H1N1)pdm09 isolates formed a single genetic cluster, belonging to the genetic subclade 6B.1, defined by HA amino acid substitutions S84N, S162N, and I216T. Furthermore, all isolates showed an oseltamivir-sensitive genotype, suggesting that Tamiflu (Oseltamivir) could still be the drug of choice in Turkey.     

Effect on cellular and humoral immune responses of the AS03 adjuvant system in an A/H1N1/2009 influenza virus vaccine administered to adults during two randomized controlled trials

The influence of AS03_A, a tocopherol oil-in-water emulsion-based adjuvant system, on humoral and cell-mediated responses to A/California/7/2009 H1N1 pandemic vaccine was investigated. In two observer-blind studies, a total of 261 healthy adults aged 18 to 60 years were randomized to receive either AS03_A-adjuvanted H1N1 vaccine containing 3.75 μg hemagglutinin (HA) or nonadjuvanted H1N1 vaccine containing 15 or 3.75 μg HA on days 0 and 21. Hemagglutination inhibition (HI) antibody and T-cell responses were analyzed up to day 42. A first dose of AS03_A-adjuvanted vaccine (3.75 μg HA) or nonadjuvanted vaccine (15 μg HA) induced HI responses of similar magnitudes that exceeded licensure criteria (e.g., 94 to 100% with titers of ≥40). A lower response following 3.75 μg HA without adjuvant was observed (73% with titers of ≥40). Following a second dose, geometric mean HI titers at day 42 were higher for AS03_A-adjuvanted vaccine (636 and 637) relative to nonadjuvanted vaccine (341 for 15 μg HA and 150 for 3.75 μg HA). Over the 42-day period, the increase in frequency of A/H1N1/2009-specific CD4⁺ T cells was significantly higher in the adjuvanted group than in the nonadjuvanted group. There was no evidence of correlation between baseline CD4⁺ T-cell frequencies and day 21 HI antibody titers, while there was some correlation (R = 0.35) between day 21 CD4⁺ T-cell frequencies and day 42 HI titers. AS03_A adjuvant enhanced the humoral and CD4⁺ T-cell-mediated responses to A/H1N1/2009 vaccine. Baseline A/H1N1/2009-specific CD4⁺ T-cell frequencies did not predict post-dose 1 antibody responses, but there was some correlation between post-dose 1 CD4⁺ T-cell frequencies and post-dose 2 antibody responses.     

甲型H1N1流行性感冒疫苗接种人群的血清抗体分析

目的:了解接种甲型H1N1流行性感冒(流感)疫苗后,人群中血清抗体的变化情况,为甲型H1N1流感疫苗的接种提供依据。方法:随机采集不同年龄已接种甲型H1N1流感疫苗人群的血清,采用血凝抑制实验检测血清中甲型H1N1流感抗体的血凝抑制滴度(HI滴度),HI滴度≥1∶40判定为阳性,同时调查采样对象的甲型H1N1流感疫苗与季节性流感疫苗的接种史。结果:甲型H1N1流感抗体阳性率为57.4%(402份/700份),抗体几何平均滴度(GMT)为1∶35.6;甲型H1N1流感抗体阳性率与GMT较高的是10～30岁组人群,较低的是60岁以上的人群;接种甲型H1N1流感疫苗后30～90天,GMT水平达到高峰(1∶56);随着季节性流感疫苗接种次数的增多,人群血清中甲型H1N1流感抗体的阳性率与GMT值反而降低。结论:青少年与成人接种甲型H1N1流感疫苗的免疫效果比儿童和老年人的好;甲型H1N1流感疫苗对人群的保护作用能持续90天左右;甲型H1N1流感抗体在0～10岁组,10～30岁组人群中持续的时间比30～60岁,>60岁组人群长;多次接种季节性流感疫苗可能会影响甲型H1N1流感抗体的产生。建议对儿童和老年人开展双倍剂量甲型H1N1流感疫苗接种;甲型H1N1流感疫苗的接种时间最好在流行期前1～3月内,并且应每年接种一次。