1株H1N1亚型猪流感病毒的全基因组特征及其对小鼠的致病性

旨在了解河南省猪流感病毒的流行情况及其遗传进化和基因组特征。2018年4月，从河南省某一出现疑似流感症状猪群中采集鼻拭子样品150份用于分离病毒，对分离病毒的全基因组进行序列测定和分析。同时感染6周龄BALB/c小鼠，研究其对小鼠的致病性。结果显示，获得1株H1N1亚型病毒[命名为A/swine/Henan/NY20/2018（H1N1）]。遗传进化表明，其HA和NA基因属于欧亚类禽H1N1分支，PB2、PB1、PA、NP和M基因属于2009甲型H1N1分支，NS基因属于经典H1N1分支。HA蛋白的裂解位点序列为PSIQSR↓GL，具有低致病性流感病毒的分子特征，在小鼠肺和鼻甲有效复制并能引起肺组织病理学变化。本研究分离到1株3源重排H1N1亚型病毒，对小鼠呈现一定致病力，提示应进一步加强对SIV的监测。     

Identification of an H1N1 subtype of swine influenza virus and serological analysis

To investigate the epizootic of swine influenza virus(SIV), 60 nasal swabs were collected from a clinical cases of pig farm in Tai'an City, Shandong Province of China in April 2017. SIV was isolated by inoculating into 10-day-old Special Pathogen Free embryonated eggs and the whole genome was sequenced. An H1N1 subtype SIV was isolated and designated as A/swine/Shandong/TA04/2017(H1N1). Phylogenetic analysis showed that apart from the polymerase A(PA) fragment belonging to the 2009 pandemic H1N1 branch, seven genome segments belonged to avian-like H1N1 influenza virus lineage. The cleavage site sequence of the hemagglutinin(HA) protein was PSIQSR↓G, which is a typical molecular biological characteristic. Five potential N-glycosylation sites(N14, N26, N277, N484 and N543) were found in the HA gene. To further investigate the epidemiology of SIV in this farm, the 995 serum samples were assessed with EAH1N1 2009 pandemic H1N1 and H3 N2 antigens. The results showed that the total positive rate was 65.43%. The positive rates of single virus infection detected by EAH1N1, 2009 pdmH1N1 and H3 N2 for serum HI(Hemagglutination inhibition) were 48.35, 30.85 and 7.47%, respectively. The results showed that SIV in Shandong Province has been reassorted in some segments and the SIV-positive rate was high on the SIV outbreak farm. These data provide evidence of an epizootic of SIV.     

中国2种橡胶树炭疽病菌对咪鲜胺敏感性比较

炭疽病是橡胶树上一种重要的叶部病害,主要病原有胶孢炭疽菌和尖孢炭疽菌。为了解中国各植胶区炭疽菌对咪鲜胺的敏感性,采用生长速率法室内测定来自海南、云南、广东和广西4省(区)22株橡胶树胶孢炭疽病菌和15株尖孢炭疽菌对咪鲜胺的敏感性。结果表明:供试的37株橡胶炭疽病菌菌株对咪鲜胺的EC_(50)值为0.022 0~0.272 1 mg/L,平均为(0.061 7±0.055 4)mg/L,EC_(50)最大值是最小值的12.5倍。其中89.2%参试菌株的EC_(50)值介于0~0.100 0 mg/L。根据成组t检验分析结果表明,目前中国4个植胶区的2种橡胶炭疽病菌对咪鲜胺敏感性无明显差异。除了来自海南和云南地区的尖孢炭疽菌有显著性差异外,其他来自不同省份的胶孢炭疽菌和尖孢炭疽菌对咪鲜胺敏感性均无明显差异。研究显示中国橡胶炭疽菌对咪鲜胺仍具有较高的敏感性,但菌株间敏感性有一定的差异,存在一定的潜在抗性风险。     

脲酶抑制剂与硝化抑制剂对稻田土壤氮素转化的影响

【目的】本研究旨在阐明脲酶抑制剂(urease inhibitor,UI)和硝化抑制剂(nitrification inhibitor,NI)对稻田土壤氮素转化的影响,探讨抑制剂提高稻谷产量以及氮肥利用率的机理。【方法】本试验设在我国南方红壤稻田,共5个处理:1)不施氮肥(CK);2)尿素(U);3)尿素+脲酶抑制剂(U+UI);4)尿素+硝化抑制剂(U+NI);5)尿素+脲酶抑制剂+硝化抑制剂(U+UI+NI);脲酶抑制剂采用N-丁基硫代磷酰三胺(NBPT),硝化抑制剂采用3,4-二甲基吡唑磷酸盐(DMPP)。在水稻分蘖期和孕穗期测定土壤脲酶活性、硝酸还原酶活性、土壤铵态氮含量、硝态氮含量以及微生物碳、氮的含量,分析NBPT与DMPP对水稻两个主要生育期土壤氮素供应的影响,比较各处理的产量以及氮肥利用率,通过逐步回归分析研究以上各指标对产量的影响,探明脲酶抑制剂NBPT与硝化抑制剂DMPP在稻田的增效机理。【结果】1)与单施尿素相比,添加NBPT以及NBPT与DMPP配施均显著提高稻谷产量与地上部氮素回收率,两个处理分别增产6.56%与8.24%,氮素回收率提高幅度为19.4%与23.7%。2)与单施尿素相比,添加NBPT以及NBPT与DMPP配施,显著降低水稻分蘖期的土壤脲酶活性和铵态氮含量,显著提高孕穗期的铵态氮含量,而对此时期的脲酶活性无显著影响,所有处理对两个时期的硝态氮含量、硝酸还原酶活性、微生物量碳、氮含量均无显著影响;因此,NBPT对于抑制脲酶活性以及提高铵态氮含量的作用主要在孕穗期之前,而单施DMPP没有显著效应。3)从各项土壤指标与水稻产量相关性的逐步回归分析结果来看,水稻分蘖期与孕穗期稻田土壤中铵态氮含量对水稻产量影响显著,而且孕穗期的影响大于分蘖期,其余指标则对产量无显著影响。【结论】脲酶抑制剂NBPT以及NBPT与硝化抑制剂DMPP配施显著提高孕穗期土壤中的铵态氮含量,显著提高稻谷产量以及地上部氮素回收率,证明了生产上氮肥后移的重要意义。     

Cepharanthine induces autophagy via PI3K/AKT/mTOR signaling pathway in ovarian cancer SKOV3 cells

AIM: To investigate the autophagy of human ovarian cancer SKOV3 cells induced by cepharanthine and to explore its mechanism. METHODS: The effect of cepharanthine on the viability of ovarian cancer SKOV3 cells was measured by CCK-8 assay. The SKOV3 cells were treated with cepharanthine, and then the formation of autophagosome was observed with acridine orange staining under fluorescence microscope. The protein levels of LC3, AKT, p-AKT, mTOR, p-mTOR and GAPDH in the SKOV3 cells treated with cepharanthine were determined by Western blot.RESULTS: Cepharanthine significantly inhibited the viability of ovarian cancer SKOV3 cells in a dose-dependent manner (P<0.05). The number of the intracellular acidic autophagosomes with bright red fluorescence was significantly increased after cepharanthine treatment in the SKOV3 cells. The expression of LC3-Ⅱ in SKOV3 cells was significantly enhanced after cepharanthine treatment. Furthermore, treatment with cepharanthine in the SKOV3 cells also resulted in a significant down-regulation of phosphorylated form of AKT and mTOR (P<0.01), while the total protein level was not changed. Combination of cepharanthine and 3-methyladenine resulted in a substantial decrease in the cell viability compared with using cepharanthine alone.CONCLUSION: Cepharanthine significantly inhibits the growth of human ovarian cancer SKOV3 cells and induces the autophagy, which may be correlated with down-regulation of PI3K/AKT/mTOR signaling pathway.     

Responses of plant community to the linkages in plant-soil C:N:P stoichiometry during secondary succession of abandoned farmlands,China

Succession is one of the central themes of ecology;however,the relationship between aboveground plant communities and underground soils during secondary succession remains unclear.In this study,we investigated the composition of plant community,plant-soil C:N:P stoichiometry and their relationships during secondary succession after the abandonment of farmlands for 0,10,20,30,40 and 50 a in China,2016.Results showed that the composition of plant communities was most diverse in the farmlands after secondary succession for 20 and 50 a.Soil organic carbon and total nitrogen contents slightly decreased after secondary succession for 30 a,but both were significantly higher than those of control farmland(31.21%-139.10%and 24.24%-121.21%,respectively).Moreover,C:N ratios of soil and microbe greatly contributed to the changes in plant community composition during secondary succession of abandoned farmlands,explaining 35.70%of the total variation.Particularly,soil C:N ratio was significantly and positively related with the Shannon-Wiener index.This study provides the evidence of synchronous evolution between plant community and soil during secondary succession and C:N ratio is an important linkage between them.     

Sorghum extrusion process combined with biofortified sweet potato contributed for high iron bioavailability in Wistar rats

This study aimed to evaluate the effect of sorghum and sweet potato on the bioavailability of iron, gene expression of proteins involved in iron metabolism and the plasma antioxidant capacity in animals fed with whole sorghum grains processed by dry heat or extrusion, combined or not with sweet potato flour with high content of carotenoids. Five experimental groups were tested (n = 7): dry heat sorghum flour (DS); extruded sorghum flour (ES); whole sorghum flour + sweet potato flour (DS + SP); extruded sorghum flour + sweet potato flour (ES + SP) and positive control (FS). The evaluations included: hemoglobin gain, hemoglobin regeneration efficiency, gene expression of divalente metal transporter 1 (DMT-1), duodenal citochroma B (DcytB), ferroportin, hephaestin, transferrin and ferritin and total plasma antioxidant capacity (TAC). The ES + SP group showed higher (p < 0.05) expression of DcytB, ferroportin and hephaestin when compared to the control group. The DS group showed high (p < 0.05) expression of DMT-1 and the ES showed high mRNA expression of transferrin and ferritin. The changes in the sorghum physicochemical properties from extrusion process reduced the iron and phytate content, and increased the gene expression of proteins involved in iron metabolism, improving iron bioavailability. The combination of sweet potato and sorghum flour (dry or extruded) improved the iron capture and total antioxidant capacity, probably due to the presence of β-carotene and antioxidant compounds.     

基于GIS与RS的北方防沙屏障带生态系统格局演变

研究北方防沙屏障带生态系统格局演变特征,对加强屏障带建设、改善生态环境具有重要意义。以北方防沙屏障带为研究区,根据2005、2010、2015年MODIS遥感影像,将北方防沙屏障带生态系统类型划分为森林、草地、湿地、农田、城镇、荒漠和裸地7类。采用空间统计、转移矩阵、景观指数分析法、PNTIL模型等方法,对北方防沙屏障带2005—2015年生态系统类型时空演变特征进行分析。结果表明,2005—2015年北方防沙屏障带生态系统整体呈稳定状态;从分屏障带看,内蒙古防沙屏障带荒漠面积明显减少,塔里木防沙屏障带森林面积迅速增加,而河西走廊防沙屏障带草地面积明显增加;从正/逆转换方向来看,屏障带内农田、城镇、荒漠及裸地生态系统正向转换率高于逆向转换率。10年间,在景观水平上,北方防沙屏障带斑块数量、斑块密度呈减少趋势;在类型水平上荒漠景观整体呈现破碎化萎缩的趋势特征,森林、草地向形状简单化的方向发展。研究表明,2005—2015年,北方防沙屏障带生态总体向好,治沙效果明显,但局部仍面临较大压力。     

CRISPR/cas系统及其在家禽上的应用研究进展

CRISPR/cas是一种获得性免疫防御系统,广泛存在于细菌和古细菌中。Strep tococcus pyogenes cas9(Spcas9)作为目前研究最多、最清楚的效应蛋白,因其酶活性高和靶点广而被应用于生命科学各个领域,但Spcas9分子量大、脱靶率高等缺点在一定程度上限制了其应用。随着CRISPR系统技术被深入开发,一些新效应蛋白被发现,本文就CRISPR/cas系统分类、原理、新效应蛋白发现及cas9(S.Pyogenes)多种变构体技术开发在及家禽上的应用进行综述,为相关领域的研究提供参考。     

Effect of Kechuanning on airway remodeling and protein level of p-ERK1/2 in lung tissues of asthmatic rats induced by virus

AIM:To investigate the effect of Kechuanning on airway remodeling and the protein level of p-ERK1/2 in lung tissues of asthmatic rats induced by virus. METHODS:The asthmatic rat model induced by respiratory syncytial virus was established. The experimental rats were divided into normal group, asthma model group, low dose (0.33 mL/kg), middle dose (3.0 mL/kg) and high dose (10 mL/kg) of Kechuanning groups, and PD98059 (3 mg/kg) group. The airway responsiveness of the rats was measured by animal ventilator. The pathological changes of the lung tissues were observed by HE staining. PAS staining and Masson staining were used to observe goblet epithelial cells metaplasia and airway collagen deposition. The expression of matrix metalloproteinases-9 (MMP-9) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in the lung tissues of the rats was detected by immunohistochemical staining. The protein levels of ERK1/2 and p-ERK1/2 were determined by Western blot. RESULTS:Compared with model group, the airway responsiveness of the rats in middle dose and high dose of Kechuanning groups was significantly decreased (P<0.01), the injury of lung tissues was significantly decreased, the goblet epithelial cells metaplasia and airway collagen deposition were significantly reduced (P<0.01), and the expression of MMP-9 and TIMP-1 in the lung tissues was also significantly decreased (P<0.01). In addition, the protein level of p-ERK1/2 in high dose of Kechuanning group was significantly decreased compared with model group (P<0.01). CONCLUSION:Kechuanning may treat asthma by regulating the expression of p-ERK1/2 in the lung tissues and improving the airway remodeling symptoms of asthmatic rats induced by virus.