H5亚型禽流感DNA疫苗质粒pCAGGoptiHA5对高致病力禽流感病毒攻击的免疫保护

对H5亚型禽流感DNA疫苗质粒pCAGGoptiHA5的免疫效果进行了研究。pCAGGoptiHA5分别以100和10μg剂量一次或两次免疫3周龄SPF鸡，首次免疫后4周以同样剂量和途径进行第二次免疫，一次免疫后4周、两次免疫后2周分别用100LD50的HPAIV A／Goose／GuangDong／1／96（H5N1）鼻腔途径进行攻击，观察发病与死亡情况，分别于攻毒后第3、5、7天采集喉头及泄殖腔拭子进行病毒分离、滴定检测排毒情况，同时检测免疫后、攻毒前及攻毒后血清HI抗体、NT抗体以及AGP抗体的动态变化。结果，100μg pCAGGoptiHA5一次免疫、100μg pCAGGoptiHA5两次免疫以及10μg pCAGGoptiHA5两次免疫均可对免疫鸡形成100％完全保护（不发病、不致死、不排毒），10μg剂量pCAGGoptiHA5一次免疫可对免疫鸡形成100％的保护（不发病、不致死），结果表明，pCAGGoptiHA5作为疫苗效果良好、成本低廉，有望成为预防H5亚型高致病力禽流感的高效、安全新型基因工程疫苗。     

Familial Congenital Methemoglobinemia in Pomeranian Dogs Caused by a Missense Variant in the NADH‐Cytochrome B5 Reductase Gene

Background

In veterinary medicine, congenital methemoglobinemia associated with nicotinamide adenine dinucleotide (NADH)‐cytochrome b5 reductase (b5R) deficiency is rare. It has been reported in several breeds of dogs, but little information is available about its etiology.

Objectives

To analyze the NADH‐cytochrome b5 reductase gene, CYB5R3, in a Pomeranian dog family with methemoglobinemia suspected to be caused by congenital b5R deficiency.

Animals

Three Pomeranian dogs from a family with methemoglobinemia were analyzed. Five healthy beagles and 5 nonrelated Pomeranian dogs without methemoglobinemia were used as controls.

Methods

Methemoglobin concentration, b5R activity, and reduced glutathione (GSH) concentration were measured, and a turbidity index was used to evaluate Heinz body formation. The CYB5R3 genes of the affected dog and healthy dogs were analyzed by direct sequencing.

Results

Methemoglobin concentrations in erythrocytes of the affected dogs were remarkably higher than those of the control dogs. The b5R activity of the affected dogs was notably lower than that of the control dogs. DNA sequencing indicated that this Pomeranian family carried a CYB5R3 gene missense variant (ATC→CTC at codon 194) that resulted in the replacement of isoleucine (Ile) by leucine (Leu).

Conclusions and Clinical Importance

This dog family had familial congenital methemoglobinemia caused by b5R deficiency, which resulted from a nonsynonymous variant in the CYB5R3 gene. This variation (c.580A>C) led to an amino acid substitution (p.Ile194Leu), and Ile194 was located in the proximal region of the NADH‐binding motif. Our data suggested that this variant in the canine CYB5R3 gene would affect function of the b5R in erythrocytes.     

Effect of dasatinib in a xenograft mouse model of canine histiocytic sarcoma and in vitro expression status of its potential target EPHA2

Canine histiocytic sarcoma (HS) is an aggressive and highly metastatic tumor. Previously, the kinase inhibitor dasatinib was shown to have potent growth inhibitory activity against HS cells in vitro, possibly via targeting the EPHA2 receptor. Here, the in vivo effect of dasatinib in HS cells was investigated using a xenograft mouse model. Moreover, the expression status of EPHA2 was examined in six HS cell lines, ranging from insensitive to highly sensitive to dasatinib. In the HS xenograft mouse model, dasatinib significantly suppressed tumor growth, as illustrated by a decrease in mitotic and Ki67 indices and an increase in apoptotic index in tumor tissues. On Western blot analysis, EPHA2 was only weakly detected in all HS cell lines, regardless of sensitivity to dasatinib. Dasatinib likely results in the inhibition of xenograft tumor growth via a mechanism other than targeting EPHA2. The findings of this study suggest that dasatinib is a targeted therapy drug worthy of further exploration for the treatment of canine HS.     

The construction and application of a population physiologically based pharmacokinetic model for methadone in Beagles and Greyhounds

Methadone is an opioid analgesic in veterinary and human medicine. To help develop appropriate pain management practices and to develop a quantitative model for predicting methadone dosimetry, a flow‐limited multiroute physiologically based pharmacokinetic (PBPK) model for methadone in dogs constructed with Berkeley Madonna? was developed. The model accounts for intravenous (IV), subcutaneous (SC), and oral administrations, and compartmentalizes the body into different components. This model was calibrated from plasma pharmacokinetic data after IV administration of methadone in Beagles and Greyhounds. The calibrated model was evaluated with independent data in both breeds of dogs. One advantage of this model is that most physiological parameter values for Greyhounds were taken directly from the original literature. The developed model simulates available pharmacokinetic data for plasma concentrations well for both breeds. After conducting regression analysis, all simulated datasets produced an R ² > 0.80 when compared to the measured plasma concentrations. Comparative analysis of the dosimetry of methadone between the breeds suggested that Greyhounds had ~50% lower 24‐hr area under the curve (AUC) of plasma or brain concentrations than in Beagles. Furthermore, population analysis was conducted with this study. This model can be used to predict methadone concentrations in multiple dog breeds using breed‐specific parameters.     

Novel reassortant H5N6 highly pathogenic influenza A viruses in Vietnamese quail outbreaks

Avian influenza A H5N6 virus is a highly contagious infectious agent that affects domestic poultry and humans in South Asian countries. Vietnam may be an evolutionary hotspot for influenza viruses and therefore could serve as a source of pandemic strains. In 2015, two novel reassortant H5N6 influenza viruses designated as A/quail/Vietnam/CVVI01/2015 and A/quail/Vietnam/CVVI03/2015 were isolated from dead quails during avian influenza outbreaks in central Vietnam, and the whole genome sequences were analyzed. The genetic analysis indicated that hemagglutinin, neuraminidase, and polymerase basic protein 2 genes of the two H5N6 viruses are most closely related to an H5N2 virus (A/chicken/Zhejiang/727079/2014) and H10N6 virus (A/chicken/Jiangxi/12782/2014) from China and an H6N6 virus (A/duck/Yamagata/061004/2014) from Japan. The HA gene of the isolates belongs to clade 2.3.4.4, which caused human fatalities in China during 2014–2016. The five other internal genes showed high identity to an H5N2 virus (A/chicken/Heilongjiang/S7/2014) from China. A whole-genome phylogenetic analysis revealed that these two outbreak strains are novel H6N6-like PB2 gene reassortants that are most closely related to influenza virus strain A/environment/Guangdong/ZS558/2015, which was detected in a live poultry market in China. This report describes the first detection of novel H5N6 reassortants in poultry during an outbreak as well as genetic characterization of these strains to better understand the antigenic evolution of influenza viruses.     

南果梨周年干物质与氮磷钾积累动态

【目的】明确南果梨干物质积累特征和氮磷钾养分周年动态积累规律,为南果梨优化施肥量和施肥时期提供依据。【方法】以12年生南果梨树为试材,采用田间采样和树体分解方法,分别于萌芽后10 d(萌芽期)、 30 d(花期)、 65 d(幼果膨大期)、 100 d(果实膨大或新稍停止生长期)、 130 d(果实着色前)、 155 d(果实采收期)、 185 d(采收后)、 210 d(落叶前)8个生育期,选干周和树高一致的3株树,将树体连根从土壤中挖出,分出果实、 叶片、 枝条、 主干、 主根、 侧根、 须根,各器官单独称重,并取200 g左右的鲜样按清水、 洗涤剂、 清水、 1%盐酸、 3次去离子水冲洗、 杀青、 烘干后,电磨粉碎过0.15 mm筛,测定样品中氮、 磷、 钾含量。【结果】1)南果梨周年生育期内,树体干物质当年净积累量为18.4 kg/plant,干物质累积速率出现两次累积高峰,分别是幼果膨大期(0.15 kg/d)和采收期(0.11 kg/d)。2)南果梨树体总氮周年积累量为154.0~301.0 g,新生器官为0~116.2 g,果实膨大期达到最高;多年生器官氮积累量为154.0~194.8 g,落叶前达到最高。3)南果梨树体总磷周年积累量为17.1~37.2 g,果实着色前最高。其中新生器官为13.7 g,果实采收期最高;多年生器官为17.1~24.9 g,果实转色期最高。4)南果梨树体总钾周年积累量为27.9~174 g。新生器官钾为97.3 g,采收期最高;多年生器官钾为27.6~76.6 g,落叶前最高。5)产量大约为17 t/hm2的12年生南果梨从萌芽到落叶前树体当年氮磷钾的单株净累积量分别为146.2、 20.1、 146.1 g,折合1000 kg果实经济产量需吸收氮(N)、 磷(P)、 钾(K)5.4、 0.7、 5.4 kg。【结论】南果梨周年干物质单株积累总量为41.4 kg,当年净积累量为19.7 kg。南果梨干物质积累主要集中在花期到果实膨大期和果实转色到落叶前,分别占47.3% 和47.5%。南果梨从萌芽到落叶前氮、 磷、 钾的单株净累积量分别为146.2、 20.1、 146.1 g,每1000 kg果实经济产量需吸收氮(N)、 磷(P)、 钾(K)5.4、 0.7、 5.4 kg。从开花到果实膨大期和从果实着色到采收后30天对氮吸收分别占总氮累积量的39.0%和49.0%,而磷、 钾的累积从萌芽到开花较快,到果实膨大期磷的累积达67.4%,钾的累积达65.1%,果实膨大期是干物质和氮磷钾积累的关键时期。     

基于纵向传导的蔬菜批零价格关系研究——以北京市果类蔬菜为例

从蔬菜产业的纵向看,批发和零售是两个重要环节,蔬菜批发价格和零售价格的走势以及它们之间的关系是各界关注的焦点。本文以北京市场为例,以番茄和黄瓜两种果类蔬菜的按日批零价格为对象,运用有限分布滞后模型和误差修正模型进行实证分析,研究蔬菜批零价格间的传导关系,并在此基础上揭示蔬菜批发价格在蔬菜价格形成中的作用。研究发现:蔬菜批零价格走势基本一致,相关系数很高;批零价格间存在批发价格向零售价格单向传导的因果关系;批零价格的传导是顺畅的,但不同种蔬菜传导的时间效应有所不同;批发价格在蔬菜价格形成中占据主导作用,当蔬菜价格受到冲击时,若零售价格过高则其会向着批发价格下调,若批发价格过高则零售价格会随之走高。     

高校心理健康教育网络课程模式的构建与实例分析

计算机多媒体技术和网络技术日新月异的发展,教育信息化的推进,为努力实现高校心理健康教育课程教学内容、教学手段和教学方法的全面创新,本文以《心理健康教育》网络课程为例,尝试剖析其在模式构建、教学设计、交互安排、教学应用等方面的特征,以期为开辟高校心理健康教育课程的新模式提供思路。     

小兴安岭天然林不同树种的树高曲线模型

选用10种树高曲线模型作为候选模型,以决定系数(R~2)、残差平方和(S_(se))和均方差(M_(se))作为模型优劣的评价指标,对小兴安岭天然林中的云杉(Picea asperata)、红松(Pinus koraiensis)、水曲柳(Fraxinus mandschurica)、椴树(Tilia tuan)4种树种的树高曲线模型进行优选。结果表明:云杉、红松和椴树的最优模型均为Logistic模型,水曲柳的最优模型为抛物线模型;云杉和红松两种针叶树种,最优模型的优势不明显,水曲柳和椴树两种阔叶树种最优模型的优势较明显;比较R~2的大小,得出较适合小兴安岭云杉和红松两种针叶树种的树高曲线模型(R~20.8),对小兴安岭阔叶树种的适用程度不具有普遍性,对椴树的适用性好(R~2最大达0.94),对水曲柳的适用性较差(R~20.8)。对水曲柳的树高进行分段研究得出:水曲柳在树高低于9 m时,树高曲线模型较适宜(R~20.8),其中抛物线模型为最优模型;树高较高时,没有适用的树高曲线模型。