美国SL-9甘薯茎尖脱毒组织培养的最适激素浓度配比初探

为探索高淀粉甘薯品种美国SL-9茎尖脱毒组织培养的最适激素浓度配比,选择6-BA和α-NAA两种激素,通过不同浓度的配比,利用线性综合评价模型对愈伤组织诱导率L、出芽(成苗)率M和芽长(苗高)N三个培养效果指标进行综合评价。结果表明,最适于该品种茎尖脱毒组织培养的培养基配方为MS+1.00 mg/L6-BA+0.50 mg/Lα-NAA;在此培养基上,美国SL-9愈伤组织诱导率L为83.3%,出芽(成苗)率M为100%,且成苗速度快,培养时间短,接种培养40d苗高达11.4mm,优于其它激素浓度配比。     

H3、H9亚型禽流感病毒双重荧光定量PCR方法的建立及初步应用

虽然H3、H9亚型禽流感病毒（Avian influenza virus, AIV）是低致病性禽流感病毒,但其具有跨宿主感染哺乳动物的能力,对社会公共卫生安全具有重大的潜在危害,因而对这些亚型病毒的监测极其重要。通过比对GenBank上H3、H9亚型禽流感病毒HA基因序列,设计了分别针对H3 AIV和H9 AIV的特异性探针。特异性试验和标准曲线试验的结果显示,这两个探针仅分别特异性识别H3、H9亚型AIV,且具有较好的扩增效率。通过重组质粒pMD19-T-H3HA和pMD19-T-H9HA 10倍梯度稀释液为模板,进行敏感性实验。结果表明,本研究所建立的荧光定量PCR方法能检测到100个DNA拷贝数H3 AIV和10个DNA拷贝数的H9 AIV,比传统PCR方法敏感性分别提高了100倍和1000倍。此外,本方法还能检测到10 EID50的H3亚型AIV或H9亚型AIV,比传统PCR方法检测敏感性均提高了1000倍。批间和批内试验的变异系数均小于3%,表明本研究建立的方法具有较好的重复性。此外,与传统PCR方法相比,用H3、H9亚型AIV双重荧光定量PCR方法检测人工感染动物的肺组织时,针对H3 AIV的敏感性提高了10~100倍,针对H9 AIV的敏感性提高了100倍。综上所述,本研究所建立的H3、H9亚型禽流感病毒双重荧光定量PCR方法具有较高的特异性和敏感性,对H3、H9亚型禽流感病毒监测具有重要的应用价值。     

H9亚型禽流感病毒钦州株HA基因的克隆与遗传进化

采集钦州活禽交易市场的鸡气管和泄殖腔的棉拭予样品,用H9亚型分型引物进行PCR初步筛选,阳性样品经SPF鸡胚尿囊腔接种分离病毒,通过RT-PCR方法扩增HA基因,并将其克隆到pMD—18T载体后进行序列测定和分析。结果表明,获得1株H9亚型禽流感病毒命名为：A／Chicken／Guangxi／qz40／2009（简称qz40）。测序结果表明qz40的HA基因片段全长1683bp,编码560个氨基酸;序列同源性比较结果表明,该毒株与参考毒株的核苷酸序列同源性为82．8％．99．9％,推导氨基酸同源性为87．5％．99．6％;HA基因的裂解位点氨基酸顺序为RSSRIGLF,为低致病性毒株;含有7个潜在的N-糖基化位点,其中5个位于HAl部分、2个位于HA2部分;基于H9亚型HA基因的进化树分析表明,qz40株属于欧亚种系的A／Chicken／Beijing／1／94（Ck／Bei—like）群系。     

Effect of idazoxan on permeability of blood-brain barrier and expression of MMP-9/TIMP-1 in mouse experimental autoimmune encephalomyelitis

AIM：To study the effect of idazoxan (IDA) on the permeability of blood-brain barrier (BBB) and the expression of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) in mouse experimental autoimmune encephalomyelitis (EAE).METHODS：Female C57BL/6 mice (n=36) were randomly divided into control group, EAE group and IDA group, with 12 mice in each group. EAE was induced by myelin oligodendrocyte glycoprotein 35-55 (MOG35-55). IDA (2 mg/kg, ip, bid) was administered for 15 d after immunization. The neurological defects of the mice were observed daily and scored. The pathological changes were observed under microscope with HE staining and LFB myelin staining. The BBB permeability was detected by Evans blue extravasation. The expression of MMP-9 and TIMP-1 in the brain of EAE mice was determined by Western blotting.RESULTS：Compared with EAE group, the score of neurological defects in IDA group was decreased, the inflammation was relieved, the BBB permeability was reduced, and the expression MMP-9 and the ratio of MMP-9/TIMP-1 were decreased (P<0.05).CONCLUSION：The neuroprotective effect of IDA on mouse EAE might be related to the down-regulation of MMP-9 and the ratio of MMP-9/TIMP-1, thus reducing the degradation of BBB and the permeability of BBB, and ameliorating the pathologic process of EAE.     

Effect of Raptor on invasion ability of glioma cells

AIM：To study the influence of Raptor on the invasion ability of glioma cells. METHODS：The technique of RNA interference was used. U87 cells were transfected with Raptor restricted siRNA plasmid, and the expression level of Raptor in the transfected cells was detected by Western blotting. The invasive ability of the cancer cells in vitro was determined. The phosphorylation level of ARK5 and the expression of MMP-2 and MMP-9 were detected by Western blotting. The expression levels of Raptor in the tumor samples of low-grade gliomas (WTO grade I and grade II) and high-grade gliomas (WTO grade III and grade IV) were also analyzed by immunohistochemical staining. RESULTS：Raptor siRNA was transfected into U87 cells and the cells were named siRaptor/U87 cells. The cells transfected with the control plasmid was named Scr/U87 cells. The expression level of Raptor in siRaptor/U87 cells was lower than that in Scr/U87 cells. The results of in vitro invasion assay showed that the number of siRaptor/U87 cells penetrating the Matrivgel matrix membrane was less than that of Scr/U87 cells (P＜0.01). The protein expression of MMP-2 and MMP-9, and phosphorylation of ARK5 protein in the cells in the experimental group were lower than those in control group. The correlation between the expression of Raptor in gliomas and the degree of deterioration was also observed (P＜0.01). CONCLUSION：The expression of Raptor may contribute to the invasion ability of glioma cells by phosphorylation of ARK5 and increase in the levels of MMP-2 and MMP-9.     

人工催熟日本鳗鲡精子的显微和超微结构

对经人工催产获得的日本鳗鲡（Anguilla japonica）精子的显微和超微结构进行了观察。光镜下观察，日本鳗鲡精子有两种形态结构，一种精子的细胞核为圆形或近 似圆形，这精子较小，细胞核长径为(2.57±0.62) μm，短径为(2.11±0.59) μm，鞭毛长度为(38.35±7.71) μm；另外一种精子的细胞核为“眉形”或“新月形”，精子较大，细胞核长径为(7.66±1.09) μm，短径为(2.54±0.46) μm，鞭毛长度为(38.35±9.02) μm。透射电镜观察结果显示：圆形精子头部的顶端无顶体，植入窝位于细胞核底端的中间，由细胞核向内凹陷而成，呈一沟状，其走向与精子的长轴平行。中心粒复合体位于植入窝内。细胞核的下端有2～3个线粒体。基体的头端呈筒状，由电子致密物构成。基体的尾端分裂成9束。尾部从袖套腔中伸出且细长，轴丝始端与基体的尾端相连，微管多呈电子致密状，轴丝为典型的“9+2”结构。圆形精子为日本鳗鲡正常成熟的精子。“眉形”或“新月形”精子头部细胞核弯曲，在弯曲处有一圆形的球状物，球状物内含有线粒体和中心粒复合体，轴丝为“9+0”结构。这种精子可能存在发育缺陷或没有达到完全正常成熟。     

H7N9亚型禽流感对辽宁省畜牧业的影响及其对策

禽流感已成为国际上广泛关注的公共卫生事件,因此,加强对流感状况的认识和了解将有助于人们有效地防控流感。本文对禽流感病毒的流行现状及对辽宁省畜牧业的影响进行综述,旨在为有效防制禽流感提供理论依据。     

Novel human H7N9 influenza virus in China

Outbreaks of H7N9 avian influenza in humans in 5 provinces and 2 municipalities of China have reawakened concern that avian influenza viruses may again cross species barriers to infect the human population and thereby initiate a new influenza pandemic. Evolutionary analysis shows that human H7N9 influenza viruses originated from the H9N2, H7N3 and H11N9 avian viruses, and that it is as a novel reassortment influenza virus. This article reviews current knowledge on 11 subtypes of influenza A virus from human which can cause human infections.     

Helminth parasites of freshwater fishes from Cuatro Ciénegas,Coahuila, in the Chihuahuan Desert of Mexico: inventory and biogeographical implications

As part of an ongoing inventory of the helminth parasites of freshwater fishes in Mexico, 570 individual fish were collected between Apr 2008 and Oct 2011 in 26 localities along the Cuatro Ciénegas region in Coahuila State, northern Mexico. Seventeen species of hosts, mostly corresponding to Nearctic freshwater elements, were studied. A total of 8324 individual worms were collected during this survey, representing 25 species of helminths, of which 9 were digeneans, 3 monogeneans, 3 acanthocephalans, 9 nematodes and 1 cestode. Most of the records in this checklist represent new host or locality records. The information provided in this checklist may be helpful for our understanding of the biodiversity and historical biogeography of this host–parasite system, because in the Cuatro Ciénegas region occur a Nearctic freshwater fish fauna, along with Neotropical and endemic elements, and from a biogeographical point of view, this may represent a transitional area.