含锂生物玻璃在大鼠心肌样细胞中的生物相容性研究

目的制备含锂生物玻璃(Li/BGs)纳米材料, 并探讨其对大鼠心肌样细胞(H9C2)和人类脐静脉内皮细胞(HUVECs)的生物相容性。方法采用溶胶-凝胶法, 将0.7 g十六烷基三甲基溴化铵、10 ml乙酸乙酯、7 ml 1 mol/L氨水、3.6 ml正硅酸四乙酯、3.57 g四水硝酸钙和0.37 g碳酸锂反应至少4 h, 制备Li/BGs。扫描电子显微镜(SEM)观察Li/BGs纳米颗粒的尺寸和形态。H9C2和HUVECs分别置于10～1 000 μg/ml浓度的Li/BGs培养基中, 细胞计数试剂盒(CCK-8)细胞毒性实验(450 nm吸光度值)评估细胞活性。细胞在10 μg/ml浓度的Li/BGs溶液中培养24 h后, 采用鬼笔环肽染色法(Phalloidin)和4’, 6-二脒基-2-苯基吲哚(DAPI)染色, 观察细胞核(蓝色)和细胞骨架蛋白(红色)的共聚焦显微镜图像, 评估Li/BGs对细胞形态的影响。组间比较采用单因素方差分析分析数据统计学差异。结果 SEM观察显示, 制备的Li/BGs颗粒近球形, 粗糙且均匀, 直径(82.86±14.28) nm。H9C2经钙黄绿...     

不同浓度异丙酚对体外人辅助性T淋巴细胞分化的影响

目的 探讨不同浓度异丙酚对体外人辅助性T淋巴细胞(Th细胞)分化的影响.方法 健康志愿者20名,男性9名,女性11名,年龄20～50岁,体重50～70 kg,每名健康志愿者分别采集外周静脉血20ml,进行全血培养,采用随机区组设计,分别接受下述处理:空白对照组(C组)、不同浓度异丙酚组[P1组(1 μg/ml)、P2组(4 μg,ml)、P3组(16 μg/ml)]、不同浓度异丙酚赋形剂组[I1 组(0.1μl/ml)、I2组(0.4μl/ml)、I3组(1.6 μl/ml)].每份血样培养于无菌24孔培养板中,依次加入上述药物培养24 h后加入氟波醇乙酯、离子霉素及蛋白质转运抑制剂刺激4 h,采用三色流式细胞技术检测全血中Th1细胞及Th2细胞的亚群比例,并计算Th1细胞与Th2细胞比值(Th1/Th2).结果 与C组比较,P3组Th1细胞亚群比例和Th1/Th2升高,I3组Th2细胞亚群比例降低(P＜0.05),其余组各指标差异无统计学意义(P＞0.05);与P1组比较,p3 组Th1细胞亚群比例和Th1/Th2升高(P＜0.05);与I3组比较,p3 组Th1细胞、Th2细胞的亚群比例升高(P＜0.05).结论 异丙酚16 μg/ml可使Th细胞向Th1细胞分化增加,1.4 μg/ml对Th细胞分化无影响.     

丹参多酚酸盐预处理对肾小管上皮细胞凋亡的保护机制

目的：为丹参多酚酸盐治疗肾脏急性肾损伤提供理论依据.方法：将肾小管上皮NRK52E细胞随机分组.对照组不干预;H2O2组加入H2O2使终浓度为400 μmol/L,丹参多酚酸盐组分为1、2、3组分别加入丹参多酚酸盐使终浓度分别为0.4 μg/ml、4 μg/ml、40 μg/ml,H2O2＋丹参多酚酸盐1、2、3组分别加入H2O2及丹参多酚酸盐,检测SOD数值,观察细胞损伤情况,MTT法观察细胞的活性,流式细胞术检测细胞凋亡率,RT-PCR检测Survivin,基因mRNA的表达.结果：H2O2组细胞凋亡率明显高于对照组及丹参多酚酸盐组;H2O2＋丹参多酚酸盐组凋亡率明显低于H2O2组;H2O2＋丹参多酚酸盐组Survivin mRNA表达明显高于H2O2组.结论：丹参多酚酸盐能显著抗氧化,从而抑制H2O2诱导的肾小管上皮细胞凋亡,其作用机制可能与其可增加Survivin表达有关;丹参多酚酸盐可用于肾脏急性损伤的治疗.     

氯胺酮对人Th细胞分化及其转录因子GATA-3和T-bet表达的影响

目的 观察氯胺酮对体外培养的人Th细胞分化的过程及其核蛋白转录因子GATA-3和T-bet表达的影响.方法 利用体外细胞培养技术建立人Th细胞培养体系,根据加入药物的不同分为六组:对照组(C组)、氯胺酮50 μg/ml组(K组)、氯胺酮50 μg/ml+N-甲基-D门冬氨酸(NM-DA)50 μg/ml组(KN1组)、氯胺酮50 μg/ml +NMDA 100 μg/ml组(KN2组)、氯胺酮50 μg/ml+NMDA 150 μg/ml组(KN3组)、NMDA 150 μg/ml组(N组).淋巴细胞经过PHA刺激48 h后,检测淋巴细胞中Th1细胞、Th2细胞亚群及其比值(Th1/Th2)、培养上清液中干扰素(INF)-γ和白细胞介素( IL)-4的含量及淋巴细胞中核蛋白转录因子GATA-3和T-bet的表达.结果 与C组比较,K、KN1、KN2和KN3组Th1、Th2细胞亚群减少,但K、KN1组Th1/Th2升高(P＜0.05或P＜0.01);与K组比较,KN3组Th1、KN2和KN3组Th2细胞亚群增多,KN2和KN3组Thl/Th2降低(P＜0.05或P＜0.01).与C组比较,K、KN1、KN2和KN3组IFN-γ和IL-4含量均降低(P＜0.05或P＜0.01);与K组比较,KN2和KN3组IFN-γ和IL-4含量增高(P＜0.05或P＜0.01).K、KN1、KN2和KN3组GATA-3和T-bet表达明显低于C组,且KN2和KN3组GATA-3和T-bet表达高于K组(P＜0.01),并且呈现剂量依赖性.结论 氯胺酮通过作用于NMDA受体对Th1和Th2细胞的分化产生抑制作用,这种抑制作用是通过抑制核蛋白转录因子GATA-3和T-bet的表达来实现的.     

GATA-3和T-bet与吗啡抑制成人辅助性T淋巴细胞分化的关系

目的 评价核蛋白转录因子GATA-3和T-bet与吗啡抑制成人辅助性T淋巴细胞(Th细胞)分化的关系.方法 健康志愿者10名,年龄20 ～ 50岁,体重50 ～ 70 kg,清晨空腹采集外周静脉血样20 ml,分离外周血单个核细胞(PBMCs).采用随机配伍设计,将每份血样分离出的PBMCs随机分为5组(n=10):常规培养组(C组)、刺激剂刺激分化组(P组)、吗啡50 μg/ml组(M组)、吗啡50μg/ml+纳洛酮50 μg/ml组(MN组)、纳洛酮50μg/ml组(N组).刺激剂孵育4h后分别收集细胞和上清液.应用流式细胞术检测Th1、Th2细胞亚群比例,计算二者比值(Th1/Th2);用酶联免疫吸附法检测上清液中干扰素(IFN)-γ和白细胞介素(IL)-4的浓度;使用凝胶电迁移技术检测淋巴细胞核蛋白转录因子GATA-3和T-bet的活性.结果 与P组比较,M组Th1、Th2细胞亚群比例、Th1/Th2、上清液IFN-γ和IL-4的浓度、GATA-3和T-bet活性降低,MN组Th1细胞亚群比例、Th1/Th2、上清液IFN-γ浓度降低(P ＜0.05或0.01);与M组比较,MN组Th1、Th2细胞亚群比例、Th1/Th2、上清液IFN-γ和IL-4的浓度、GATA-3和T-bet活性升高(P＜0.05或0.01);N组与C组上述各指标比较差异无统计学意义(P＞0.05).结论 吗啡通过激活阿片受体抑制Th细胞分化,而这种抑制作用与抑制核蛋白转录因子GATA-3和T-bet的活性有关.     

自体肿瘤疫苗治疗结直肠癌72例分析

近年来自体肿瘤疫苗主动特异性免疫治疗已成为预防和治疗恶性肿瘤复发和转移的一种新的有效手段[1]。自1996年开展该项目以来,我们已成功应用该技术治疗各类恶性肿瘤共381例,其中结直肠癌72例,现报告如下。1资料和方法1.1一般资料1996-1999年大坪医院行结直肠癌根治性切除的患者共198例。治疗组72例加用了自体肿瘤疫苗治疗,男性47例,女性25例,年龄41～65岁,平均53.5岁,其中高、中及低分化腺癌各为19、31及22例。单纯手术组126例仅行常规治疗。1.2自体肿瘤疫苗的制备将切除的肿瘤标本放入含50μg/ml庆大霉素的生理盐水中,无菌条件下快速送至…     

静脉注射利多卡因对丙泊酚抑制人工流产术扩张宫颈时体动半数有效浓度的影响

目的：探究静脉注射利多卡因对丙泊酚施耐德（Schnider）模型靶控输注（TCI） 抑制人工流产术扩张宫颈时体动半数有效浓度（EC₅₀）的影响。
方法：选择自愿要求终止妊娠的初次人工流产患者46例,年龄 18～35岁,妊娠6～9周,BMI 18～25 kg/m²,ASA Ⅰ或Ⅱ级。根据计算机生成的随机序列分为两组:利多卡因组（L组,n=25）和生理盐水组（C组,n=21）。L组静脉注射利多卡因1 mg/kg,C组静脉注射等容量的生理盐水。采用Schnider模型TCI丙泊酚,初始效应室靶浓度设为4.0 μg/ml,依据Dixon序贯法,如在扩张宫颈时发生2级及以上体动反应,为阳性反应,则下一例升高一个浓度梯度,否则降低一个浓度梯度,相邻浓度梯度为0.5 μg/ml。连续出现7个阳性反应拐点时终止研究。采用Probit回归分析计算两组丙泊酚抑制人工流产术扩张宫颈时体动的EC₅₀和95%有效浓度（EC₉₅）及其95%可信区间（CI）。记录意识消失时间、苏醒时间、丙泊酚使用量和宫颈扩张效果,丙泊酚注射痛和术后15 min VAS疼痛评分,围术期低氧血症、低血压、术后恶心呕吐和头晕等不良反应的发生情况。
结果：L组丙泊酚抑制人工流产术扩张宫颈时体动的EC₅₀为3.82 μg/ml（95％CI 3.59～4.03 μg/ml）,EC₉₅为4.25 μg/ml（95％CI 4.04～5.10 μg/ml）。C组丙泊酚抑制人工流产术扩张宫颈时体动的EC₅₀为4.09 μg/ml（95％CI 3.71～4.41 μg/ml）,EC₉₅为4.65 μg/ml（95％CI 4.36～6.48 μg/ml）。与C组比较,L组丙泊酚EC₅₀明显降低,丙泊酚使用量明显减少,宫颈扩张总有效率明显升高,丙泊酚注射痛和低氧血症发生率明显降低（P＜0.05）。
结论：静脉注射利多卡因1 mg/kg可降低Schnider模型TCI丙泊酚抑制人工流产术扩张宫颈时体动的EC₅₀,减少丙泊酚使用量,降低静脉注射痛发生率,宫颈扩张效果好,不良反应少。     

2009年杭州市甲型H1N1流感监测现状分析

目的探讨2009年杭州市甲型H1N1流感的流行规律、临床表现和病原学特征。方法对2009年杭州市13家流感监测哨点医院的每日门诊流感样病例和甲型H1N1流感确诊病例的流行病学进行分析。采用RT—PCR方法对咽拭子中血凝素（HA）基因进行扩增和测序,并用DNASTAR软件进行序列分析。全文数据采用SPSS12．0软件进行统计分析,率的比较采用χ^2检验。结果流感样病例监测显示,第28周之前,疫情处于平稳状态,之后全市流感样病例逐渐增多。第35周时达到高峰,全市流感样病例就诊比例为7．47％（5442／72859）。之后疫情回落,但到第41周时又再次回升,至第46周出现第二次高峰,全市流感样病例就诊比例达到11．32％（11034／97436）。第38周以前,杭州市人群中主要流行株是季节性H1N1型,其次有少量的乙型和季节性流感H1N1型,到第44周,甲型H1N1流感成为唯一的优势流行株,主要易感人群是11～25岁的青少年,以学生居多。序列分析显示,2009年杭州市甲型H1N1流感病毒株与北美株、中国其他地区的毒株以及疫苗株同源性达到99％,关键位点高度保守,但与季节性流感病毒株同源性只有70％。结论2009年杭州市流感疫情发展迅速,人群中流行的优势毒株为甲型流感H1N1型,易感人群为青少年,目前未发现变异株及高致病株,但仍需进一步监测。     

NR2B多肽疫苗对神经病理性痛大鼠的镇痛效应

目的 评价NR2B多肽疫苗对神经病理性痛大鼠的镇痛效应.方法 成年雌性SD大鼠,体重180～200 g,制备保留性神经损伤(SNI)模型,7 d后选取SNI模型制备成功的大鼠18只,随机分为3组(n=6),NR2B组沿背部两侧皮下多点注射NR2B多肽疫苗50 μl(含NR2B 100μg)3次,每隔2周注射1次;PBS组和KLH组分别给予PBS 50 μl和匙孔虫戚血蓝蛋白100 μg,均用弗氏佐剂稀释至100 μl.分别于制备SNI前1 d(基础值)、第1次免疫前1 d及第3次免疫后14d时测定机械痛阈;分别于制备SNI前1 d及第3次免疫后14 d时取血清和脑脊液,测定NR2B抗体滴度,然后处死大鼠,取L3-5脊髓组织,其中3只采用免疫组化法测定脊髓背角胶质纤维酸性蛋白表达水平,反映星形胶质细胞激活程度;另外3只采用Western blotting法测定脊髓背角NR2B蛋白表达水平.结果 NR2B组第3次免疫后14 d时血清及脑脊液中均可检测到NR2B抗体,而SNI前1 d时未检测到NR2B抗体,PBS组及KLH组各时点均未检测到NR2B抗体.与基础值比较,各组机械痛阈均降低(P<0.05);与PBS组和KLH组比较,NR2B组第3次免疫后机械痛阈升高,脊髓背角星形胶质细胞激活程度降低,NR2B蛋白表达下调(P<0.05).结论 NR2B多肽疫苗对神经病理性痛大鼠具有镇痛作用,其机制与进入脊髓的NR2B抗体下调了NR2B蛋白表达有关.     

人工流产术患者复合异丙酚时靶控输注瑞芬太尼的药效学

目的 探讨人工流产术患者复合异丙酚4.5 μg/ml时靶控输注瑞芬太尼的药效学.方法 拟行人工流产术患者135例,年龄18～30岁,ASAI级,孕6～10周.随机分为9组(n=15):瑞芬太尼效应室靶浓度分别为0.5、0.8、1.1、1.4、1.7、2.0、2.3、2.6和2.9 ng/ml(Ⅰ组～Ⅸ组).各组异丙酚效应室靶浓度均为4.5 μg/ml.采用概率单位回归分析法,计算麻醉效果达优时瑞芬太尼效应室靶浓度EC50、EC95及其95%可信区间(CI)和呼吸抑制时瑞芬太尼效应室靶浓度EC50、EC95及其95%CI.结果 麻醉效果达优时瑞芬太尼的效应室靶浓度EC50为1.67 ng/ml,其95%CI为1.45～1.90 ng/ml,EC95为3.88 ng/ml,其95%CI为3.08～5.89 ng/ml;呼吸抑制时瑞芬太尼效应室靶浓度EC50为2.44 ng/ml,其95%CI为2.28～2.64 ng/ml,EC95为3.36 ng/ml,其95%CI为2.99～4.34 ng/ml.麻醉效果达优时瑞芬太尼的效应室靶浓度EC95高于呼吸抑制时效应室靶浓度EC95(P＜0.05).结论 人工流产术患者复合异丙酚4.5 μg/ml时,麻醉效果达优时瑞芬太尼的效应室靶浓度EC50、EC95,分别为1.67、3.88 ng/ml,呼吸抑制时瑞芬太尼的效应室靶浓度EC50、EC95,分别为2.44、3.36 ng/ml.     

Expression and Characterization of HA1 Protein of Highly Pathogenic H5N1 Avian Influenza Virus for Use in a Serodiagnostic Assay

The hemagglutinin ectodomain (HA1 subunit) from highly pathogenic avian influenza (HPAI) isolate (A/chicken/Vietnam/14/2005) was cloned and expressed using a baculovirus expression vector. Biosynthesis, glycosylation and secretion of the HA1 proteins, with natural or a melittin signal peptide at the N‐terminus and a six‐histidine (6xHis) tag at the C‐terminus, were examined in insect cells. A 40‐kDa unglycosylated precursor and a fully processed, mature form of the HA1 protein migrated around 52 kDa were detected by SDS‐PAGE and confirmed by Western blot using H5N1‐specific antibody. Treatment of tunicamycin and peptide‐N‐glycosidase F (PNGase F) further revealed that the recombinant HA1 proteins produced in insect cells were indeed glycosylated with N‐linked oligosaccharide side chains. Time‐course experiments showed that substitution of the HA natural sequence with the signal sequence from honeybee melittin promoted a high level of expression and efficient secretion of the HA1. A high yield, 37 μg/ml, of HA1 protein was obtained from recombinant baculovirus‐infected cell culture supernatant. In addition, the cell surface expression of rHA1 was detected by indirect immunofluorescent staining and showed biological activity on hemadsorption assays. Recombinant HA1 protein‐based ELISA was evaluated and appeared to be sensitive and specific for the rapid detection of H5 subtype‐specific antibodies in serum samples. No cross‐reactivity to antibodies from 15 other influenza A subtypes was detected. Taken together, the newly developed recombinant HA1‐based ELISA could offer an alternative to other diagnostic approaches for the specific detection of H5 avian influenza virus infection.     

Influenza virus vaccination in kidney transplant recipients: serum antibody response to different immunosuppressive drugs

Salles MJC, Sens YAS, Boas LSV, Machado CM. Influenza virus vaccination in kidney transplant recipients: serum antibody response to different immunosuppressive drugs.
Clin Transplant 2010: 24: E17–E23. © 2009 John Wiley & Sons A/S.   Abstract: 
Introduction:  This study prospectively accessed the immune response to the inactivated influenza vaccine in renal transplant recipients receiving either azathioprine or mycophenolate mofetil (MMF). Side effects were investigated.
Methods:  Sixty-nine patients received one dose of inactivated trivalent influenza vaccine. Antihemagglutinin (HI) antibody response against each strain was measured before and one to six months after vaccination.
Results:  Geometric mean HI antibody titers for H1N1 and H3N2 strains increased from 2.57 and 2.44 to 13.45 (p = 0.001) and 7.20 (p < 0.001), respectively. Pre- and post-vaccination protection rates for H1N1 and H3N2 increased from 8.7% to 49.3% (p < 0.001); and 36.3% (p < 0.001) and seroconversion rates were 36% and 25.3%, respectively. There was no response to influenza B. The use of MMF reduced the H1N1 and H3N2 protection rates and the seroconversion rate for the H1N1 strain when compared with the use of azathioprine, and subjects transplanted less than 87 months also had inferior antibody response. Adverse events were mild and there were no change on renal function post-vaccination.
Conclusion:  Renal transplant patients vaccinated against influenza responded with antibody production for influenza A virus strains, but not for influenza B. Use of MMF and shorter time from transplantation decreased the immune response to the vaccine.     

Evaluation of Screening Assays for the Detection of Influenza A Virus Serum Antibodies in Swine

Increased surveillance of influenza A virus (IAV) infections in human and swine populations is mandated by public health and animal health concerns. Antibody assays have proven useful in previous surveillance programmes because antibodies provide a record of prior exposure and the technology is inexpensive. The objective of this research was to compare the performance of influenza serum antibody assays using samples collected from pigs (vaccinated or unvaccinated) inoculated with either A/Swine/OH/511445/2007 γ H1N1 virus or A/Swine/Illinois/02907/2009 Cluster IV H3N2 virus and followed for 42 days. Weekly serum samples were tested for anti‐IAV antibodies using homologous and heterologous haemagglutination‐inhibition (HI) assays, commercial swine influenza H1N1 and H3N2 indirect ELISAs, and a commercial influenza nucleoprotein (NP)‐blocking ELISA. The homologous HIs showed 100% diagnostic sensitivity, but largely failed to detect infection with the heterologous virus. With diagnostic sensitivities of 1.4% and 4.9%, respectively, the H1N1 and H3N2 indirect ELISAs were ineffective at detecting IAV antibodies in swine infected with the contemporary influenza viruses used in the study. At a cut‐off of S/N ≤ 0.60, the sensitivity and specificity of the NP‐blocking ELISA were estimated at 95.5% and 99.6%, respectively. Statistically significant factors which affected S/N results include vaccination status, inoculum (virus subtype), day post‐inoculation and the interactions between those factors (P < 0.0001). Serum antibodies against NP provide an ideal universal diagnostic screening target and could provide a cost‐effective approach for the detection and surveillance of IAV infections in swine populations.     

Natural influenza infection produces a greater diversity of humoral responses than vaccination in immunosuppressed transplant recipients

The humoral immune response to influenza virus infection is complex and may be different compared to the antibody response elicited by vaccination. We analyzed the breadth of IgG and IgA responses in solid organ transplant (SOT) recipients to a diverse collection of 86 influenza antigens elicited by natural influenza A virus (IAV) infection or by vaccination. Antibody levels were quantified using a custom antigen microarray. A total of 120 patients were included: 80 IAV infected (40 A/H1N1 and 40 A/H3N2) and 40 vaccinated. Based on hierarchical clustering analysis, infection with either H1N1 or H3N2 virus showed a more diverse antibody response compared to vaccination. Similarly, H1N1-infected individuals showed a significant IgG response to 27.9% of array antigens and H3N2-infected patients to 43.0% of antigens, whereas vaccination elicited a less broad immune response (7.0% of antigens). Immune responses were not exclusively targeting influenza hemagglutinin (HA) proteins but were also directed against conserved influenza antigens. Serum IgA responses followed a similar profile. This study provides novel data on the breadth of antibody responses to influenza. We also found that the diversity of response is greater in influenza-infected rather than vaccinated patients, providing a potential mechanistic rationale for suboptimal vaccine efficacy in this population.     

Pandemic 2009 influenza H1N1 virus vaccination: compliance and safety in a single hemodialysis center

Background: Acceptance of vaccination against pandemic 2009 H1N1 influenza virus has been poor in some countries, perhaps because of concerns about safety of the new vaccine. Subjects and methods: We prospectively examined vaccination compliance and reasons for nonvaccination in the dialysis patients and the staff of a single hemodialysis unit, after on-site vaccination with a monovalent inactivated adjuvanted H1N1 vaccine. Safety profile was evaluated and compared to that of a control group without chronic kidney disease (CKD). Results: Vaccination acceptance among dialysis patients in our unit was 68% (110/161). Dialysis patients vaccinated against H1N1 had significantly higher compliance with vaccination for seasonal influenza and pneumococcus than those unvaccinated. Three out of 34 (9%) of the unit's staff received the vaccine. Fear of side effects was the main reason for not vaccinating in both groups, while several participants did not consider pandemic influenza a serious disease. At least one adverse reaction was observed in 37/110 (33.6%) of the vaccinated dialysis patients and in 22/42 (52.4%) of the non-CKD control group, p = 0.034. Local mild pain at injection site was the most common side effect of vaccination in both groups. All side effects were of short duration and no serious adverse reactions related to the vaccine or reactions of special interest occurred during the follow-up period. Conclusions: Our findings indicate that immunization against H1N1 virus in dialysis patients is safe and do not support the concerns about safety of the vaccine that was the main reason for nonvaccination in our study.     

Unusually High Mortality in Waterfowl Caused by Highly Pathogenic Avian Influenza A(H5N1) in Bangladesh

Mortality in ducks and geese caused by highly pathogenic avian influenza A(H5N1) infection had not been previously identified in Bangladesh. In June–July 2011, we investigated mortality in ducks, geese and chickens with suspected H5N1 infection in a north‐eastern district of the country to identify the aetiologic agent and extent of the outbreak and identify possible associated human infections. We surveyed households and farms with affected poultry flocks in six villages in Netrokona district and collected cloacal and oropharyngeal swabs from sick birds and tissue samples from dead poultry. We conducted a survey in three of these villages to identify suspected human influenza‐like illness cases and collected nasopharyngeal and throat swabs. We tested all swabs by real‐time RT‐PCR, sequenced cultured viruses, and examined tissue samples by histopathology and immunohistochemistry to detect and characterize influenza virus infection. In the six villages, among the 240 surveyed households and 11 small‐scale farms, 61% (1789/2930) of chickens, 47% (4816/10 184) of ducks and 73% (358/493) of geese died within 14 days preceding the investigation. Of 70 sick poultry swabbed, 80% (56/70) had detectable RNA for influenza A/H5, including 89% (49/55) of ducks, 40% (2/5) of geese and 50% (5/10) of chickens. We isolated virus from six of 25 samples; sequence analysis of the hemagglutinin and neuraminidase gene of these six isolates indicated clade 2.3.2.1a of H5N1 virus. Histopathological changes and immunohistochemistry staining of avian influenza viral antigens were recognized in the brain, pancreas and intestines of ducks and chickens. We identified ten human cases showing signs compatible with influenza‐like illness; four were positive for influenza A/H3; however, none were positive for influenza A/H5. The recently introduced H5N1 clade 2.3.2.1a virus caused unusually high mortality in ducks and geese. Heightened surveillance in poultry is warranted to guide appropriate diagnostic testing and detect novel influenza strains.     

Detection of reassortant avian influenza A (H11N9) virus in wild birds in China

Human infectious avian influenza virus (AIV) H7N9 emerged in China in 2013. The N9 gene of H7N9, which has the ability to cause death in humans, originated from an H11N9 influenza strain circulating in wild birds. To investigate the frequency and distribution of the N9 gene of the H11N9 and H7N9 influenza virus circulating in wild birds between 2006 and 2015, 35,604 samples were collected and tested. No H7N9 but four strains of the H11N9 subtype AIV were isolated, and phylogenetic analyses showed that the four H11N9 viruses were intra‐subtype and inter‐subtype reassortant viruses. A sequence analysis revealed that all six internal genes of A/wild bird/Anhui/L306/2014 (H11N9) originated from an H9N2 AIV isolated in Korea. The H9N2 strain, which is an inner gene donor reassorted with other subtypes, is a potential threat to poultry and even humans. It is necessary to increase monitoring of the emergence and spread of H11N9 AIV in wild birds.     

Heterologous Immune Responses to Influenza Vaccine in Kidney Transplant Recipients

Influenza vaccine is known to have suboptimal immunogenicity in transplant recipients. Despite this, influenza vaccine may have the added benefit of inducing a cross‐reactive immune response to viral strains not found in the vaccine. This is termed “heterologous immunity” and has not been assessed previously in transplant patients. Pre‐ and postvaccination sera from kidney transplant recipients (n = 60) immunized with the 2012–2013 adjuvanted or nonadjuvanted influenza vaccine underwent testing by hemagglutination inhibition assay for strains not present in vaccine: A/New Caledonia/20/99 (H1N1), A/Texas/50/2012 (H3N2) and B/Brisbane/60/2008. The geometric mean titer of antibody to heterologous strains increased after vaccine (H1N1: 80.0 to 136.1, p < 0.001; H3N2: 23.3 to 77.3, p < 0.001; B: 13.3 to 19.5, p < 0.001). Seroconversion rates were 16.7%, 41.7%, and 13.3%, respectively. No differences in heterologous response were seen in the adjuvanted versus nonadjuvanted groups. Patients were more likely to seroconvert for a cross‐reactive antigen if they seroconverted for the specific vaccine antigen. Seroconversion to heterologous A/H3N2, for example, was 84.0% for homologous H3N2 seroconverters versus 11.4% for nonseroconverters (p < 0.001). This study provides novel evidence that transplant recipients are able to mount significant cross‐protective responses to influenza vaccine that may be an additional, previously unknown benefit of immunization.