鲤春病毒血症病毒Shlj1株糖蛋白空间结构及其B细胞抗原表位的预测

为预测鲤春病毒血症病毒(Spring Viremia of Carp Virus,SVCV)Shlj1株糖蛋白(Glycoprotein,G)的空间结构及B细胞抗原表位,采用RT-PCR法从接种SVCV的鲤上皮细胞悬液提取的病毒总RNA中扩增糖蛋白基因,并通过序列测定获得Shlj1株糖蛋白基因的核苷酸序列及氨基酸序列,利用Phyre2对Shlj1分离株的糖蛋白空间结构进行预测,使用DNAStar软件综合分析糖蛋白的柔性区域、亲水性、表面可能性和抗原指数参数。结果表明:获得的SVCV Shlj1株糖蛋白核苷酸序列长1527 bp,推导出其氨基酸序列为509aa;空间结构预测显示,SVCV Shlj1株糖蛋白存在一定数量的α-螺旋、β-折叠、β-转角及大量无规则卷曲结构,空间构象较规则;抗原指数及抗原表位指数分析显示,糖蛋白中存在许多抗原指数较高的区域,该区域平均抗原指数为1.025,最大值达1.250,其中5个区域(4~26、145~157、293~302、315~327、407~491氨基酸区段)可能是B细胞抗原表位的主要分布区域。本研究结果为SVCV糖蛋白表位疫苗的设计及单克隆抗体的制备奠定了理论基础。     

江西地区儿童感染的乙型肝炎病毒全基因组序列分析

目的：初步调查江西地区乙肝疫苗免疫儿童感染的乙型肝炎病毒全基因组的分子特征。方法收集江西省儿童医院11445份乙肝疫苗免疫儿童血清标本,采用酶联免疫吸附试验检测 HBV-M,抽提乙肝表面抗原（HB-sAg）阳性血清中的 HBV DNA,分3个相互重叠片段 PCR 扩增 HBV 基因并测序,采用 ContigExpress 软件对所测序列拼接成全基因组序列,分析各读码框核苷酸和氨基酸变化。对 HBV 不同基因型以及所获得的 HBV 全基因组的 S 基因序列进行系统进化树的构建以确定基因型。结果11445份血清样品中,检测出117份 HBsAg 阳性标本,进行分段重叠扩增并测序,最终拼接出58份完整的 HBV 基因组,均为 B 基因型和 adw 血清型。对各开放读码框进行序列比对发现 PreS 区存在散在分布的突变：如 A2990C→ H48P、A2999C→ N51T、C3041T→ P65L、C3097A→L84I 和 A3136G→ T97A;S 区突变集中在120～200 aa,主要突变位点为 C533A→ P127T、A541C→Q129H、G588C→G145A、T753A→F200Y;前 C 区突变位点有 A1814C／T1815C→M1P、G1896A→ W28＊,C 区出现1例 C1913A→P5T 变异;P 区出现散在分布的突变位点,未发现与耐药性有关的突变位点,其 RT 区 YMDD 核序也未发生变异;X 区氨基端主要突变点为：G1437T／A→ G22C／S、G1476A→ G35R 和 G1503A／T／C／T1504G→V44I／F／L／C,羧基端突变集中在120～150 aa,出现5例 A1762T／G1764A→K130M／V131I 突变。结论了解分析儿童 HBV 全基因组的分子特征和突变情况,为疫苗免疫儿童感染乙肝的预防和治疗提供病毒学分子信息。     

基于T7启动子表达系统的呼吸道合胞病毒微型基因组的拯救

目的:以T7启动子表达系统为基础,通过RNA病毒反向遗传学操作,拯救人呼吸道合胞病毒(human respiratory syncytial virus,RSV)微型基因组。方法:构建可分别表达RSV四种核壳体蛋白的辅助质粒px8δT-PT1-N,px8δT-PT1-P,px8δT-PT1-M2-1和px8δT-PT1-L以及含增强型绿色荧光蛋白(enhanced green fluorecent protein,EGFP)开放读码框(open reading frame,ORF),RSV病毒前导序列(leader region/genomic promotor),转录起始信号(gene start,GS)、转录终止信号(gene end,GE)和尾随序列(trailer region/antigenomic promoter)等顺式作用元件(cis-acting elements)的微型基因组重组质粒pSC11-E,在上述的5个质粒中,均引入T7 RNA多聚酶(T7 RNA polymerase,T7 RNP)启动子,经鉴定正确后,先以pSC11-E转染RSV感染的可组成型表达T7多聚酶的BSR T7/5细胞进行拯救,确定微型基因组编码质粒设计的合理性,而后通过5个质粒共转染BSR T7/5细胞,并通过荧光显微镜观察荧光的表达情况判断拯救成功与否。结果:成功构建了基于T7启动子表达系统的RSV微型基因组5质粒拯救系统,并实现了拯救。结论:该系统的成功构建,有助于今后对RSV病毒开展基因修饰研究。     

两起新型布尼亚病毒感染病例的病毒序列分析

目的:分析发热伴血小板减少综合征布尼亚病毒感染(SFTSV)感染患者的临床特点、流行病学以及SFTSV基因序列。方法:收集本市2014年4-6月某医院送检的2例重症发热伴血小板减少(SFTS)患者的血清标本,经核酸荧光PCR检测阳性确诊。对病毒核酸测序并与GenBank数据比对。结果:两名患者均有典型的SFTSV感染的临床症状,以发热、腹泻为首发症状,血常规见粒系及血小板呈进行性降低。从2例患者血清提取的核酸经测序,其基因序列与GenBank中的SFTSV进行比对,0428株(安庆)的RNA多聚酶、糖蛋白、NS蛋白的同源性与安徽代表株AHL/China/2011的同源性最高,均达到99%。0507株(滁州)上述目的基因与江苏株JS2011-013-1的同源性高达99%。结论:SFTSV在本地区散发存在,基因序列分析结果提示其来源或有差异。     

没食子酸对恩诺沙星在鲤体内的药代动力学及残留的影响研究

为研究没食子酸与恩诺沙星联合用药及单用恩诺沙星在福瑞鲤Cyprinus carpio体内的药代动力学参数及其残留消除规律,采用平行对照试验,将体质量为(200±30)g的鲤随机分为单用恩诺沙星组(对照)和联合用药组(没食子酸与恩诺沙星质量比为10:1),试验水温为26℃,各组均按2 mg/kg(体质量)剂量的恩诺沙星单次混饲口灌给药,于给药后不同时间点采集鲤血浆、肌肉、肝胰脏、肾脏组织,采用高效液相色谱法测定各组织中恩诺沙星的浓度,并使用DAS 3.0药物代谢动力学软件的非房室模型统计矩方法分析药时数据。结果表明,没食子酸能显著增大鲤各组织对恩诺沙星的吸收效率,提高有效药物浓度20.0%~44.9%,并加快了恩诺沙星的代谢和消除,240 h时联合用药组鲤血浆、肌肉组织中药物残留量比对照组分别降低了44.2%和34.1%。     

山西省红色旅游经典景区客流量互动影响——基于ARDL模型的实证分析

分析景区客流量之间的互动影响关系，有助于景区发展。以山西四个红色旅游经典景区麻田八路军总部纪念馆、八路军太行纪念馆、太原解放纪念馆、山西昔阳县大寨景区的客流量为研究对象，构建ARDL模型，分析四个景区之间客流量互动影响关系。结果显示：4个景区客流量之间存在互动影响关系，但影响大小和方向不同；一个景区的当期客流量不仅受自身滞后期客流量的影响，还会受其他景区当期和滞后期客流量的影响。     

从日本战时“国民组织化”反思其国民战争责任

第二次世界大战期间,日本的"国民组织化"这种战争动员形式,曾为二战时期德、意、日等法西斯国家的共有现象,但日本的国民组织化又有其自身的特点,为战时日本国民在对外侵略战争中起着重要的作用。日本必须反思侵华战争中日本国民的战争责任。     

Xenobiotic and endobiotic induction of mixed function monooxygenase in carp Cyprinus carpio

3-Methylcholanthrene (3-MC) significantly induces aryl hydrocarbon (benzo[a]pyrene) hydroxylase (AHH) activity in mature and juvenile Cyprinus carpio of both sexes. The degree of induction of AHH by 3-MC in mature fish is significantly lower than that in juveniles and induction in mature males is significantly higher than in mature females. Pretreatment with sex steroids (testosterone in males and estradiol in females) significantly reduces 3-MC-inducibility of AHH in both mature and juvenile fish, while gonadectomy in mature fish elevates AHH inducibility. AHH induction is slightly decreased by hydrocortisone and increased by metyrapone pretreatment in mature fish only. Gonadectomy further elevates metyrapone affected 3-MC inducibility of AHH in C. carpio.     

Influence of potassium permanganate on chronic toxicity of cadmium to common carp,Cyprinus carpio,and aquatic ecosystem

Common carp Cyprinus carpio was chronically exposed to cadmium (2.5 mgL^‐1) and combined treatment of cadmium and KMnO₄ (1 mgL^‐1) for 90d in outdoor artificial enclosures. Although total and filtrable (dissolved) cadmium of water gradually decreased during the experiment whole body concentration of cadmium in common carp progressively increased in both treatments. Combination of KMnO₄ with cadmium marginally reduced the accumulation of cadmium during the initial periods (30 d) but failed to produce any impact on accumulation at the end of experiment (90 d). Cadmium treatment alone did not reduce the length and weight of fish, but there was abnormality in the length weight relationship. KMnO₄ alone and in combination with cadmium significantly reduced the length, weight and yield of fish. Limnological parameters were not affected by KMnO₄ alone, but combined treatment of Cd and KMnO₄ reduced the zooplankton and phytoplankton population, primary productivity of water and NPP/CR ratio. Judicious use of KMnO₄ in aquaculture to avoid toxic interaction with cadmium has been recommended.     

池塘养殖锦鲤体表溃疡病的组织病理学观察

为临床诊断和防治锦鲤Cyprinus carpio koi皮肤溃疡病,采用病理解剖方法对患病锦鲤进行临床观察和组织病理学分析,试验用患病锦鲤取自上海市朝阳锦鲤养殖场发病池塘,确定病鱼10尾(编号1~10),取每条病鱼的组织进行病理解剖、石蜡切片、H-E染色。结果表明:患病锦鲤体表溃疡病处的病灶组织出现严重淤血、炎性细胞浸润并聚集、肌束溶解和坏死等典型病理表现;体表溃疡病是由致病菌局部感染所致,并未全身性感染,10尾病料的脾组织均表现出血管扩张、管壁增厚、血管周围的实质组织凝固变性,伴有红细胞坏死和凝固变性区域内部分组织坏死等较为一致的病理特征,可以诊断为血液中有毒物质引起的脾组织病变,导致脾功能低下;肝胰组织和肾组织较脾组织病变相对较轻;肠、心、鳃、头肾组织等基本正常。研究表明,本病例全部样品均有不同程度的脾凝固变性、肝组织淤血、肾组织间质充血及其组织中血管扩张、管壁增厚的病理变化,这与体表溃疡病变无直接关联。     

Correlations of nonylphenol-ethoxylates and nonylphenol with biomarkers of reproductive function in carp (Cyprinus carpio) from the Cuyahoga River

Various chemical and biological measures were determined in carp (Cyprinus carpio) sampled from seven sites along the Cuyahoga River, Ohio; from the relatively pristine headwaters to the lower portion heavily polluted from various industrial, urban and wastewater treatment plants (WWTP). Levels of nonylphenol (NP), nonylphenol ethoxylates (NPEs; NP1EO, NP2EO) and total NPEs (NP plus the NPEs) in fish increased in a downstream direction, with maximal values observed below the discharge of the Akron WWTP. In female fish there were no significant differences between sites in GSI or levels of vitellogenin (VTG) and 17beta-estradiol (E2). However, differences were observed between sites using measures of 11-ketotestosterone (11-KT) and the ratio E2/11-KT. In male fish the highest levels of VTG were observed downstream of the Akron WWTP and a significant correlation (r=85%) between levels of NP and VTG was demonstrated. No site differences were observed in the measures of GSI, E2, 11-KT or the E2/11-KT ratio in male fish. These data suggest that endocrine active chemicals, such as, NP and NPEs are impacting fish downstream of the Akron WWTP; however, further work is warranted to separate linkages to other possible chemical factors in the water.     

饲料中不同添加剂组合对锦鲤生长和体色的影响

为研究饲料中不同添加剂组合对锦鲤Cyprinus carpio(初始体质量为195.88 g±0.98 g)生长和体色的影响,并与日本优质锦鲤商品饲料进行增重和增色效果对比,进行了增重和增色两部分试验,其中增重部分以三色锦鲤为研究对象,探讨了试验增重饲料与商品增重饲料对其生长的影响,增色部分以红白锦鲤为研究对象,探讨了试验对比饲料、试验增色饲料和商品增色饲料对其体色的影响,各组均设3个重复,试验周期为60 d,每隔15 d测定锦鲤的生长和体色指标。结果表明:添加L-肉碱、血球蛋白粉、螺旋藻和枯草芽孢杆菌的试验饲料对三色锦鲤的促生长效果与商品增色料无显著性差异(P>0.05),但试验饲料具有更低的饲料系数;添加虾青素、螺旋藻和枯草芽孢杆菌的试验饲料对红白锦鲤体色的增艳效果与商品增色料无显著性差异(P>0.05);投喂两种试验增色料和商品增色料的锦鲤体表各部位对色素的沉积程度相似。研究表明,采用不同添加剂组合的两种试验饲料与日本优质锦鲤饲料的功效水平相当,能够分别满足对锦鲤快速增重和增色的要求。     

Plasma vitellogenin levels and gonadal morphology of wild carp (Cyprinus carpio L.) in a receiving rivers downstream of sewage treatment plants

A field study was carried out in order to assess, in a wild cyprinid fish, the biological effects related to the exposure to estrogenic compounds. The gonadosomic index (GSI), plasma vitellogenin (VTG) level and histological observations of gonads were performed to evaluate the reproductive health in common carp (Cyprinus carpio) from three rivers receiving sewage treatment plant (STP) effluents (Guadarrama, Henares and Jarama rivers, Spain) and from a reference site (Lozoya river). No significative differences in male GSI between sites were described. Only in the 18% of fish from Guadarrama river were elevated plasma VTG levels detected. The presence of oocytes in the testis was not found in any of males examined. The results suggest that the released estrogenic compounds by STPs effluents are not enough capable to adversely affect the gonadal development of the fish studied. Additional studies will be required to assess the risk of estrogenic compounds on fish from other sites of this area.     

饲料浮萍水平对黄金锦鲤生长性能、消化酶活力及抗氧化能力的影响

为研究浮萍Lemna minor对黄金锦鲤Cyprinus carpio生长性能、抗氧化和消化能力的影响,以初始体质量为(52.99±1.95)g的黄金锦鲤为研究对象,在饲料中分别添加0、3%、6%、9%、13%、14%的浮萍,相应替代同等水平的菜粕,制成6组等氮(粗蛋白质为38.5%~38.6%)、等能(12.2 MJ/kg)的饲料,分别记为D1(对照)、D2、D3、D4、D5、D6饲料组,将540尾健康的黄金锦鲤随机分成6组,每组设3个重复,每个重复放30尾鱼,养殖时间为8周。结果表明:D6饲料组增重率(WGR)、蛋白质效率(PER)、特定生长率(SGR)和肥满度(CF)均显著高于对照组及添加水平为3%的饲料组(P<0.05),而其饲料系数(FCR)显著低于上述两组(P<0.05);D6饲料组蛋白酶活力与脂肪酶活力显著高于其他饲料组(P<0.05),各饲料组间淀粉酶活力则无显著性差异(P>0.05);D4与D6饲料组肝胰脏、脾脏、肾脏和血清中的超氧化物歧化酶(SOD)与过氧化氢酶(CAT)活力显著高于其他饲料组(P<0.05),且D6饲料组丙二醛(MDA)活力显著最低(P<0.05)。研究表明:本试验条件下,饲料中浮萍的添加水平在14%时,有利于黄金锦鲤的生长,并能提高鱼体抗氧化能力与消化酶活力。