High efficient expression of human endostatin in E.coli and its antiangiogensis activity

AIM: To examine the expression of human endostatin in E.coli, produce its fusion protein antibody and observe its biological activity. METHODS: Endostatin gene was amplified by polymerase chain reaction,recombined with plasmid vector pGEX-2T and induced expression with IPTG.The protein activity was tested by endothelial cell proliferation inhibitory assay.Inclusion body crudely purified was used to generate polyclonal antibody to detect its expression at mouse's liver and kidney etc. RESULTS: The protein expressed was 20kD after digestion by thrombin,it appeared the anti-angiogenesis activity and Western blotting indicated the expression of endostatin in liver and kidney of mouse. CONCLUSION: The successful expression of human endostatin and the preparation of polycolonal antibody indicated its potential application in anti-angiogenesis therapy and diagnosis tumors.     

草莓悬浮细胞原生质体培养再生愈伤组织

以草毒品种宝交早生的花药愈伤组织建立的悬浮细胞系为试材,对原生质体分离和再生进行研究。结果表明:酶液的渗透压、浓度和配比对悬浮细胞原生质体产量和活力有重要的影响。悬浮细胞在CPW+1.0％ Cellulase R-10+0.5 Macerozyme R-10+0.05％Pectolyase Y-23+0.6mol/L甘露醇+0.5％PVP的酶液中酶解12h,原生质体产量和活力最高,分别达16.35×10~6/g和84.6％。采用液体浅层培养法对原生质体进行培养获得了再生愈伤组织。     

不同引物对植原体的检测灵敏度比较研究

用常规PCR和巢式PCR对目前常用的各种植原体引物(fPl/rP7、fU5/rP7、fU5／rU3、fAY/rEY和fFD9/rFD9)的检测灵敏度进行了比较研究。研究发现，它们的检测灵敏度并不相同。最灵敏的引物(fAY/rEY)要比最不灵敏的引物(fFD9/rFD9)的灵敏度至少高出数千倍。因此，按照不同的检测目的选择所用的检测引物是明智的，特别是当待测样品中的植原体浓度极低时，比如葡萄组织样品中植原体的检测。据此提出了用于不同检测目的时的最佳引物。     

长期施肥条件下菜田—蔬菜生态系统变化的研究( Ⅳ) 蔬菜生态系统的变化

 以2002 年设施番茄栽培为代表计算其N、P、K投入量与携出量之间的表观平衡状态, 其结果基本上和14.5 年后的土壤实测值与设施栽培本底值之间纵向比较的增长关系一致, 前者只能说明系统养分变化的趋势, 后者才是实际的养分盈亏状态, 养分盈亏主要取决于有机肥的施用与养分元素的供给。5个代表性处理模拟了蔬菜施肥的不同现状, 从土壤肥力及产量等方面的变化真实反映出各自的系统变化及发展趋势, 在本试验条件下以ANK施肥下的系统最优。对有机肥施用、化学肥料的配施以及测土施肥等问题进行了探讨。     

Virulence traits of avian pathogenic (APEC) and fecal (AFEC) <Emphasis Type="Italic">E. coli</Emphasis> isolated from broiler chickens in Algeria

Avian pathogenic E. coli (APEC) is the etiologic agent of avian colibacillosis, the most common disease responsible for chicken morbidity in the world. Although multiple virulence-associated factors were identified, their prevalence in Algeria is still poorly known. In the present research, 92 avian pathogenic E. coli (APEC) isolates were recovered from broilers with clinical signs and lesions of colibacillosis. In addition, 32 E. coli isolates collected from feces of healthy birds (AFEC) were included for comparison. All isolates were investigated by PCR for the presence of a total of 11 virulence-associated genes described for avian pathogenic (iroN, ompT, hlyF, iss, iutA, and fimC) and diarrheagenic E. coli (eae, stx, elt/est, ipaH, and aggR). The sensitivity of 39 APEC isolates to 16 antibiotics was also determined using antimicrobial pretreated microplates. Here, we report that 98% of the examined isolates host at least one of the tested virulence factors. The most prevalent genes in APEC were iutA (90.6%), ompT (86.9%), and iss (85.8%); whereas, iutA (78.1%), fimC (78.1%), and iroN (68.7%) were the highest prevalent genes in AFEC. Our data showed that none of the AFEC isolates harbor any of the tested diarrheagenic genes. Moreover, only elt/est (5.4%), stx (2.1%), and ipaH (2.1%) genes were carried by APEC isolates. We further established that ceftazodime, ceftiofur, mequindox, amoxicillin/clavulanic acid, and meropenem were the most efficient antibiotics against the analyzed APEC isolates. Overall, our findings provide more insights about APEC and AFEC virulence potential in Algeria which could participate in the fight against colibacillosis.     

PCR鉴定沙门氏菌血清分型方法的建立与应用

为建立快速有效、经济适用的沙门氏菌血清型鉴定方法,利用Primer Plex软件设计畜禽中常见沙门氏菌的菌体(O)抗原和鞭毛抗原(H)的引物,优化引物比例和退火温度等反应条件,建立沙门氏菌血清型鉴定的PCR法,并对46株沙门氏菌分离株进行血清型鉴定,同时与传统玻片凝集法和美国CDC的液相芯片(Luminex-x MAP)法进行比较。优化后获得了O抗原中B、C1、D和E1群的多重PCR引物,以及鞭毛抗原H1相抗原基因(fli C)和H2相抗原基因(flj B)的2对引物;建立了一套基于沙门氏菌O和H抗原的多重PCR血清型鉴定法;PCR法和Luminex-x MAP法鉴定出7种沙门氏菌血清型,传统玻片凝集法鉴定出6种血清型;PCR法与传统玻片凝集法和Luminex-x MAP鉴定结果的符合率均为95.65%。本研究建立的PCR沙门氏菌血清型鉴定法简便、快速,预期可在沙门氏菌的血清分型研究中发挥作用。     

肉鸡屠宰加工过程中沙门菌污染定量风险评估

为了构建肉鸡屠宰加工过程中沙门菌污染定量风险评估模型,分析鸡肉中沙门菌的消长变化规律,明确关键风险防控点,采用2015年部分肉鸡屠宰场沙门菌污染监测和调研数据,构建以烫煺后为评估过程起点,包括净膛、清洗预冷和分割传送的模块化过程风险评估模型,以不同分布描述各变量,并通过@RISK软件模拟运行。结果显示,通过构建的模型模拟屠宰加工后终端鸡肉产品中的沙门菌污染量,90%的可能分布在0~9MPN之间,而依据实际监测数据换算为5.3 MPN,说明模型可信。根据过程中各环节的模拟数据,分析了肉鸡携带的沙门菌消长变化,发现预冷后沙门菌污染总量明显下降,但分割传送后污染总量又有所回升。通过拟合的相关系数分析,明确了传送带上的沙门菌是影响终端鸡肉沙门菌污染的最关键风险点。本研究构建的肉鸡屠宰加工全过程沙门菌定量风险评估模型,可为肉鸡屠宰场沙门菌污染的卫生监管和风险管理提供理论依据。     

内蒙古羊屠宰场致泻性大肠杆菌和沙门氏菌污染状况分析

为了解内蒙古地区羊屠宰场各屠宰环节致泻性大肠杆菌和沙门氏菌的携带情况,评估是否存在污染风险,选取不同规模屠宰场,用棉拭子法分别采集不同屠宰环节样品349份,36 h内进行试验;应用PCR方法分离鉴定病原菌,对鉴定为致泻性大肠杆菌的分离株进行16S r DNA测序并构建系统发育树。结果显示:从刚宰杀的羊胴体表面和环境中共分离到致泻性大肠杆菌12株,其中11株携带毒力基因elt,1株携带毒力基因eae和elt;大中型屠宰场(1.21%)的致泻性大肠杆菌阳性率低于小型屠宰场(5.43%),不同规模屠宰场的预冷间均未检出致泻性大肠杆菌和沙门氏菌;从屠宰环境中分离出1株携带inv A基因的沙门氏菌,阳性率为0.29%;经高压冲洗和排酸预冷等屠宰工艺后,在羊胴体表面均未发现致泻性大肠杆菌和沙门氏菌。     

不同地区动物源致病性大肠杆菌的分子特性与表型分析

为进一步了解致病性大肠杆菌流行状况,减轻大肠杆菌对畜牧业和公共卫生的危害,对不同时间国内外报道的动物中分离的大肠杆菌,进行毒力、血清型、耐药性和基因型等方面的比较,对比分析大肠杆菌分子特性和表型差异,从而为临床医学和公共卫生控制提供指导。     

暖地泥炭藓(Sphagnum junhuhnianum)孢子形态及发育过程探讨

描述了暖地泥炭藓(Sphagnum junhuhnianum)孢蒴从翠绿色透明状球体历经黄绿色、淡褐色、褐色、具光泽的深咖啡色球体到最后喇叭状过程中孢子的形态特征.成熟孢蒴中孢子的数量(71 631±12 751)个.成熟孢子四面体结构清晰,极面观呈圆三角形,赤道面观呈近扇形,直径(23.27±1.05)μm,均出现三裂隙结构,唇状,稍细,长为极半径的2/3,裂隙表面布满疣状颗粒物,近极端三面凹陷无褶皱,分布少量疣状颗粒物,远端面不平整,布满疣状颗粒物.