Immunization with a Live Attenuated H7N9 Influenza Vaccine Protects Mice against Lethal Challenge

The emergence of severe cases of human influenza A (H7N9) viral infection in China in the spring of 2003 resulted in a global effort to rapidly develop an effective candidate vaccine. In this study, a cold-adapted (ca), live attenuated monovalent reassortant influenza H7N9 virus (Ah01/AA ca) was generated using reverse genetics that contained hemagglutinin (HA) and neuraminidase (NA) genes from a 2013 pandemic A H7N9 isolate, A/Anhui/01/2013 virus (Ah01/H7N9); the remaining six backbone genes derived from the cold-adapted influenza H2N2 A/Ann Arbor/6/60 virus (AA virus). Ah01/AA ca virus exhibited temperature sensitivity (ts), ca, and attenuation (att) phenotypes. Intranasal immunization of female BALB/c mice with Ah01/AA ca twice at a 2-week interval induced robust humoral, mucosal, and cell-mediated immune responses in a dose-dependent manner. Furthermore, the candidate Ah01/AA ca virus was immunogenic and offered partial or complete protection of mice against a lethal challenge by the live 2013 influenza A H7N9 (A/Anhui/01/2013). Protection was demonstrated by the inhibition of viral replication and the attenuation of histopathological changes in the challenged mouse lung. Taken together, these data support the further evaluation of this Ah01/AA ca candidate vaccine in primates.     

产耐高温谷氨酰胺转胺酶菌株的快速筛选方法

目的:采用亚硝基胍(NTG)诱变结合96孔板高通量筛选方法筛选产耐高温谷氨酰胺转胺酶(MTG)的茂原链霉菌(Streptomyces mobaraensis)。方法:通过优化96孔板高通量测定MTG活性的方法、确定筛选温度和时间,建立了产耐高温MTG菌株的快速筛选方法;通过优化NTG诱变条件建立了筛选突变库;通过96孔板高通量初筛、摇瓶复筛获得了产耐高温MTG的突变株12-82,并通过摇瓶发酵对12-82所产MTG进行热稳定性分析。结果:采用2mg/ml NTG、p H8.0、60min的诱变条件获得突变株,将突变株的发酵上清液于70℃水浴7.5min,再在37℃空气浴、反应10min的条件下测定MTG活性,从5 200株突变株中筛选出5株产耐高温MTG的突变株,其中突变株12-82在50℃水浴60min以及70℃水浴1.5min的酶活残留率均比出发株高出近20%,且80℃保温2min仍有11.9%的酶活残留率。结论:利用NTG诱变结合96孔板高通量筛选的方法筛选到5株所产MTG热稳定性相对较高的突变株,其中突变株12-82在50℃、70℃和80℃的酶活残留率均有10%~20%的提高。这为高温食品加工领域所需耐高温MTG生产菌株的高效筛选提供了可行性方案。     

红壤侵蚀退化马尾松林下不同治理模式土壤化学计量特征

以南方红壤严重侵蚀退化地低效林改造、乔灌草混交、浅沟播草、封禁治理4种治理模式马尾松林地为对象,研究不同治理模式和不同土层土壤有机碳(SOC)、全氮(TN)、全磷(TP)和枯落物养分含量及其化学计量特征,为该区水土流失治理和植被恢复提供科学依据。结果表明: 不同治理模式下土壤SOC、TN、TP和枯落物P含量差异显著,土壤和枯落物养分含量均显著高于对照,改善效果均以乔灌草混交最好。土壤SOC、TN和TP含量总体上随土层增加而减少;而浅沟播草治理模式表现为,土壤SOC、TN、TP含量均随着土层增加呈先减小后增大的趋势,养分含量在5～20 cm土层达到最低值;不同治理模式下土壤C:N、C:P、N:P和枯落物C:P均存在显著差异,土壤空间变异性表现为土壤C:N>C:P>N:P。土壤C:N除乔灌草混交外,其他治理模式处于较低水平,而土壤C:P和N:P整体上较高。枯落物C:N、C:P和N:P与土壤呈相反的变化规律,低效林改造和乔灌草混交远小于对照。土壤C:N和C:P主要受控于土壤SOC含量和枯落物C含量,土壤N:P主要受控于土壤TP含量和枯落物P含量。土壤化学计量比还受土壤含水量、土壤容重、pH等因素的影响,枯落物与土壤养分之间关系密切并表现出相似的变化规律。     

北京松山天然落叶阔叶林生态系统净碳交换特征及其影响因子

森林生态系统在陆地碳循环过程中发挥着重要作用,关于温带落叶阔叶林生态系统碳平衡过程影响机制的讨论尚未统一。本研究于2019年对北京松山典型落叶阔叶林生态系统的净碳交换量(NEE)及空气温度(T_a)、土壤温度(T_s)、光合有效辐射(PAR)、饱和水气压差(VPD)、土壤含水量(SWC)、降雨量(P)等环境因子进行原位连续监测,分析松山落叶阔叶林生态系统净碳交换特征及其对环境因子的响应。结果表明: 在日尺度上,NEE生长季(5—10月)各月平均日变化均呈“U”字形变化,日间为碳汇,夜间为碳源。其他月份NEE均为正值,变化平缓,表现为碳源。在季节尺度上,NEE呈单峰曲线变化规律,全年NEE为-111 g C·m^-2·a^-1,生态系统呼吸总量(R_e)为555 g C·m^-2·a^-1,总生态系统生产力(GEP)为666 g C·m^-2·a^-1。碳吸收与释放量分别在6月与11月达到最大值。PAR是影响日间净碳交换量(NEE_d)的主导因子,二者关系符合Michaelis-Menten模型,VPD是间接影响NEE_d的主导因子,最适宜日间净碳交换的VPD范围为1～1.5 kPa。土壤温度是影响夜间净碳交换量(NEE_n)的主导因子,SWC是NEE_n的限制因子,SWC过高或过低均会对NEE_n产生抑制,最适值为0.28 m³·m^-3。     

E-cadherin和KLF 4表达对胃癌侵袭转移的作用

观察E-cadherin,Krüppel-like factor 4(KLF4)蛋白在胃癌和正常胃黏膜组织中的表达,分析其与胃癌浸润、转移的关系.应用免疫组织化学SP法检测84例手术切除的胃癌标本及对应正常胃黏膜组织中E-cadherin,KLF4蛋白的表达.各指标之间相关因素的差异性比较采用χ2检验,E-cadherin,KLF4相关性研究采用Spearman相关分析.结果显示,与正常胃组织相比,E-cadherin、KLF4蛋白在胃癌组织中均呈低表达或者缺失(分别42.9%vs.95.24%,8.3%vs 81%,P<0.05).E-cadherin、KLF4蛋白的阳性表达率与组织分级(P<0.05)、肿瘤浸润深度(P<0.05)、淋巴转移(P<0.05)明确相关.Spearman相关分析显示KLF4蛋白与E-cadherin蛋白的表达呈正相关(P<0.05).因此,E-cadherin,KLF4蛋白水平低表达可能与胃癌浸润和转移有关,而联合检测更能有效判断胃癌这一生物学行为.     

浅析潜在外来入侵物种——苹浅褐卷蛾

苹浅褐卷蛾Epiphyas postvittana原产澳大利亚,是澳大利亚西南部、新西兰、英国及美国多种经济作物的重要害虫,目前尚未在全球范围广泛分布。由于其地理分布范围窄、寄主植物广泛、形态鉴定困难、能够对农作物造成较大的经济危害以及具有遗传多样性等特点,目前已被多数国家列为检疫对象并提出严格的检疫要求。而2007年该虫在美国加州地区的定殖使得研究人员对该虫能够造成的危害程度引起了再次的关注。在我国,该虫也被列为对外检疫对象,但是相关报道相对较少。作为检疫性害虫,对其相关信息的综合掌握是进一步进行有害生物风险分析的基础和保障。本文详细描述了苹浅褐卷蛾的地理分布、寄主范围、发生为害与环境条件的相互关系等,以期为该虫进行进一步风险分析及对其的检疫措施的制定和实施提供重要的信息与依据。     

昆虫听觉相关基因的研究进展

听觉对于昆虫的求偶、同种竞争、躲避天敌以及寄生昆虫寻找寄主等方面具有非常重要的作用。早年人们对昆虫听觉系统的形态学、生理学及行为学等方面进行了广泛研究。而近年来,研究人员对昆虫听觉分子机理开展了大量研究,对昆虫听觉相关基因包括Atonal(Ato)基因、Spalt(Sal)基因等十几种基因进行了结构分析和功能研究。本文综述了国内外昆虫听觉相关基因的研究进展。     

姜黄素诱导Nrf2核转位对脂多糖引起库普弗细胞分泌炎症细胞因子的影响

目的：研究姜黄素诱导大鼠Kupffer细胞Nrf2核转位对脂多糖（LPS）引起的炎症细胞因子分泌的影响。方法：分别用10μM、20μM和30μM干预Kupffer细胞8h,诱导Nrf2核转位水平;将Kupffer细胞随机分为对照组、LPS组和干预组,对照组正常培养未加姜黄素和LPS,LPS组用10μg／mL的LPS加入Kupffer细胞培养液共同培养2h;干预组用30μM姜黄素干预8h后,余处理同LPS组。Western blot检测Nrf2核转位水平,分光光度法检测细胞MDA、GSH水平,ELISA法检测上清液TNF-α和IL-6,放免法检测IL-1β。结果：①姜黄素诱导Kupffer细胞Nrf2核转位,核转位水平随浓度增加而增高。②LPS组MDA水平较对照组显著升高（P〈0．01）,干预组MDA水平较LPS组显著降低（P〈0．01）,仍显著高于对照组（P〈0．01）。LPS组GSH水平较对照组显著降低（P〈0．01）,干预组GSH水平较LPS组显著升高（P〈0．01）,仍显著低于对照组（P〈0．01）。③LPS组上清液TNF-α,IL-1β和IL-6显著高于对照组（P〈0．01）,干预组均显著低于模型组（P〈0．01）,但显著高于对照组（P〈0．01）。结论：姜黄素通过诱导Kupffer细胞Nrf2核转位,降低LPS诱导的氧化应激损伤,抑制Kupffer细胞分泌炎症细胞因子。     

Rat bone marrow-derived schwann-like cells differentiated by the optimal inducers combination on microfluidic chip and their functional performance

Numerous researches demonstrated the possibility of derivation of Schwann-like (SC-like) cells in vitro from bone marrow stromal cells (BMSCs). However, the concentration of the induce factors were different in those studies, especially for the critical factors forskolin (FSK) and β-heregulin (HRG). Here, we used a new and useful method to build an integrated microfluidic chip for rapid analyses of the optimal combination between the induce factors FSK and HRG. The microfluidic device was mainly composed of an upstream concentration gradient generator (CGG) and a downstream cell culture module. Rat BMSCs were cultured in the cell chambers for 11 days at the different concentrations of induce factors generated by CGG. The result of immunofluorescence staining on-chip showed that the group of 4.00 μM FSK and 250.00 ng/ml HRG presented an optimal effect to promote the derivation of SC-like cells. Moreover, the optimal SC-like cells obtained on-chip were further tested using DRG co-culture and ELISA to detect their functional performance. Our findings demonstrate that SC-like cells could be obtained with high efficiency and functional performance in the optimal inducers combination.     

Characterization of the 2009 Pandemic A/Beijing/501/2009 H1N1 Influenza Strain in Human Airway Epithelial Cells and Ferrets

Background

A novel 2009 swine-origin influenza A H1N1 virus (S-OIV H1N1) has been transmitted among humans worldwide. However, the pathogenesis of this virus in human airway epithelial cells and mammals is not well understood.

Methodology/Principal Finding

In this study, we showed that a 2009 A (H1N1) influenza virus strain, A/Beijing/501/2009, isolated from a human patient, caused typical influenza-like symptoms including weight loss, fluctuations in body temperature, and pulmonary pathological changes in ferrets. We demonstrated that the human lung adenocarcinoma epithelial cell line A549 was susceptible to infection and that the infected cells underwent apoptosis at 24 h post-infection. In contrast to the seasonal H1N1 influenza virus, the 2009 A (H1N1) influenza virus strain A/Beijing/501/2009 induced more cell death involving caspase-3-dependent apoptosis in A549 cells. Additionally, ferrets infected with the A/Beijing/501/2009 H1N1 virus strain exhibited increased body temperature, greater weight loss, and higher viral titers in the lungs. Therefore, the A/Beijing/501/2009 H1N1 isolate successfully infected the lungs of ferrets and caused more pathological lesions than the seasonal influenza virus. Our findings demonstrate that the difference in virulence of the 2009 pandemic H1N1 influenza virus and the seasonal H1N1 influenza virus in vitro and in vivo may have been mediated by different mechanisms.

Conclusion/Significance

Our understanding of the pathogenesis of the 2009 A (H1N1) influenza virus infection in both humans and animals is broadened by our findings that apoptotic cell death is involved in the cytopathic effect observed in vitro and that the pathological alterations in the lungs of S-OIV H1N1-infected ferrets are much more severe.