Protection of blue shrimp (Litopenaeus stylirostris) against the White Spot Syndrome Virus (WSSV) when injected with shrimp lysozyme

In this study we found that a blue shrimp (Litopenaeus stylirostris) lysozyme gene (Lslzm) was up-regulated in WSSV-infected shrimp, suggesting that lysozyme is involved in the innate response of shrimp to this virus. Shrimp were intramuscularly injected with Lslzm protein to identify how this recombinant protein protects L. stylirostris from WSSV infection and to determine how this protein influences nonspecific cellular and humoral defense mechanisms. Higher survival rates and a lower viral load (compared with controls) were reported for shrimps that were first injected with the Lslzm protein and then infected with WSSV. In addition, the Lslzm expression level and the immunological parameters (including THC, phagocytic activity, respiratory burst activity, phenoloxidase activity and lysozyme activity) were all significantly higher in the WSSV-infected shrimp treated with the Lslzm protein, compared with the controls. These results indicate that lysozyme is effective at blocking WSSV infection in L. stylirostris and that lysozyme modulates the cellular and humoral defense mechanisms after they are suppressed by the WSSV virus.     

腺病毒介导的人巨细胞病毒UL49基因小鼠模型的建立

建立表达HCMV UL49 基因的转基因小鼠,为抗病毒药物研究提供有效的实验动物模型。本实验将UL49-GFP基因插入腺病毒穿梭质粒pDC316中,构建重组质粒pDC316-UL49-GFP,与腺病毒骨架质粒pBHGloxΔE1,3Cre 通过脂质体介导共转染293 细胞,重组产生腺病毒Ad-UL49-GFP, 经PCR和Western Blot鉴定正确后,大量扩增、纯化,制备高滴度重组腺病毒。纯化腺病毒经尾静脉注射感染小鼠,通过荧光定量PCR 和Western blot 方法,检测UL49 基因在小鼠体内组织分布和表达时相。结果显示UL49基因在小鼠的心、肝、脾、肺、肾组织均有表达,并且表达量由高到低顺序依次是：肝、脾、肾、心、肺,在腺病毒感染第3天在各靶器官表达水平较高,此后逐渐下降,第14天时仅存在肝和脾中。表明表达UL49基因的小鼠模型构建成功。小鼠模型的成功建立为下一步筛选以UL49基因为靶的抗病毒药物奠定了基础。     

尖孢镰刀菌生产蒽醌色素的液体发酵条件研究

优化了尖孢镰刀菌液体发酵生产蒽醌类红色素的发酵条件。通过单因素实验和正交优化实验,确定最佳产色素发酵培养基为：可溶性淀粉30％,(NH4)2SO4 3％,MgSO4 0.3％,KH2PO4 4％,pH 6.0。产色素最适培养条件为：初始pH6.0,装液量20％,接种量10％,吐温-80添加量1％,温度28℃,摇床转速200r/min,发酵周期120h。此条件下,色素效价即可达到8.184U/ml,比优化前提高了1.8倍。国内首次对尖孢镰刀菌所产蒽醌色素进行研究,为其进一步应用奠定基础。     

海南霸王岭自然保护区多孔菌研究(英文)

对采自海南霸王岭自然保护区的木生多孔菌约400号标本进行了鉴定共获得110种。按字母顺序列出了这些多孔菌名录;其中浅黄林氏革孔菌Lindtneria flava为中国新记录种,提供了详细描述和显微结构绘图;同时,依据文献对世界林氏革孔菌属种类进行了总结,并提供了该属所有种类的检索表。     

国产两性霉素B抢先治疗血液病患者肺部侵袭性真菌感染的疗效与安全性分析

目的观察国产两性霉素B抢先治疗血液病患者侵袭性肺部真菌感染(以下简称IFI)的疗效与不良反应。方法回顾性分析我院2004年12月～2008年12月间,45例免疫功能低下IFI患者接受国产两性霉素B治疗的临床资料。结果 45例患者均应用两性霉素B治疗两周以上,最长应用时间为14周,中位数6周。总有效率为86.7%,不良反应主要有低钾血症、发热、转氨酶升高、肾功损害、胃肠道反应,根据WHO药物毒性分级标准均为I～II级毒性。结论国产两性霉素B对于血液病患者IFI临床诊断的抢先治疗疗效肯定,不良反应可耐受。     

含氢HTK液对大鼠供心保存中心肌细胞凋亡的影响

目的:研究合氢HTK液对供心保存过程中心肌细胞凋亡的影响.方法:SD大鼠随机分4组:对照组(保存液为HTK液)、H1组(保存液为含氢浓度约0.8mM的HTK液),H2组(含氢浓度约0.4mM的HTK液),H3组(含氢浓度约0.2mM的HTK液).采用Langendorff离体大鼠心脏灌注法,心脏分别在各组保存液中冷保存(4℃)6h,Tunel染色测定心肌细胞凋亡率,荧光定量实时PCR检测凋亡相关基因Bcl-2和Bax的mRNA表达,分光光度法检测心肌组织中Caspase-3活性.结果:供心冷保存(4℃)6h后,H1、H2、H3组凋亡指数明显低于对照组.H1、H2、H3组的Bcl-2mRNA表达水平显著高于对照组(P<0.01或P<0.05);对照组的Bax mRNA水平明显高于H1、H2、H3组(P<0.01或P<0.05),对照组caspase-3活性明显高于H1、H2、H3组(P<0.01).结论:舍氢HTK明显抑制供心保存时大鼠心脏心肌细胞凋亡.     

不同时间早期肠内营养对急性重症胰腺炎患者的影响

目的:通过观察不同时间早期肠内营养对急性重症胰腺炎患者的影响,探讨急性重症胰腺炎患者早期肠内营养的最佳时机。方法:将重症急性胰腺炎患者60例分为三组:A组、B组和C组,每组20例。A组为肠外营养组予以常规治疗,B组在常规治疗的基础上通过鼻空肠管在入院3-5天内予以早期肠内营养,C组在常规治疗的基础上通过鼻空肠管在入院5天后予以早期肠内营养。抽取所有患者入院时、入院后3天、7天、14天静脉血,ELISA法测定患者血清中TNF-α、IL-6含量,并记录患者住院期间感染率、病死率、并发症发生率、住院时间、住院费用等。结果:各组患者血清中TNF-α、IL-6含量逐渐下降,B组和C组下降较A组下降明显,差异有统计学意义(P<0.05),B组和C组比较B组予以肠内营养后下降较C组明显,差异有统计学意义(P<0.05)。B组和C组感染率、病死率、并发症发生率、住院时间、住院费用较A组明显低(P<0.05),B组与C组比较感染率、病死率、并发症发生率B组明显低于C组(P<0.05)。结论:早期肠内营养可改善急性重症胰腺炎早期炎症反应,从而减低感染率、病死率、并发症发生率。在患者循环稳定条件下,越早予以肠内营养对患者的预后越有...     

氯化钴对小鼠NIH3T3细胞增殖和凋亡的影响

目的:观察氯化钴(Cocl2)对体外培养的小鼠NIH3T3细胞增殖和凋亡的影响.方法:首先利用MTT法和流式细胞术检测不同浓度Cocl2对小鼠NIH3T3细胞增殖及凋亡的影响.然后利用免疫印迹检测HIF-1α和凋亡相关蛋白Bcl-2及Bax的表达.结果:(1)MTT结果显示,低于250μ mol/L的Cocl2作用NIH3T3细胞24 h对细胞的增殖活力影响不大,当浓度增加到500μ mol/L以上时会明显抑制细胞的增殖活力.(2)流式细胞仪检测结果显示低浓度Cocl2(≤ 250μ mol/L)并不会引起NIH3T3细胞明显的凋亡和坏死,增加浓度(≥500μmol/L)会导致细胞早期凋亡和坏死增加.3)不同浓度Cocl2作用NIH3T3细胞24 h均可诱导细胞中HIF-1α表达上调,同时Bax/Bcl-2的比值增高.结论:高浓度Cocl2可以抑制小鼠NIH3T3细胞增殖,促进凋亡发生.     

不同液体复苏方式对失血性休克大鼠血流动力学的影响

目的:探讨不同液体复苏对失血性休克大鼠血流动力学的影响及机制。方法:Wistar大鼠40只,随机分为5组:假手术组、休克组、乳酸林格液(RL)复苏组、羟乙基淀粉(HES)复苏组、自身血液(BL)复苏组,每组8只,建立失血性休克大鼠模型。在失血性休克大鼠模型复制成功后1h给予对应液体复苏2h。观察并记录各组大鼠的血流动力学变化:收缩压(SBP)、舒张压(DBP)、平均动脉压(MAP)、呼吸频率(RR)和心率(HR)。结果:三组液体复苏组休克时的SBP、DBP、MAP均较休克前低,HR、RR较休克前明显加快(P<0.05)。RL复苏组各时间点SBP、DBP、MAP明显低于休克前(P<0.05)。HES组、BL组复苏各时间点SBP、DBP、MAP与休克前相近(P>0.05),但明显高于RL组。RL组各时间点RR明显快于休克前(P<0.05),HES组、BL组复苏各时间点RR与休克前相近(P>0.05),但明显低于RL组。RL组复苏60min、90min、120min的HR明显快于休克前(P<0.05),HES组、BL组苏各时间点HR同休克前相近(P>0.05),但60min、90min、120min的HR明显低于R...     

Significance of consensus CYC‐binding sites found in the promoters of both ChCYC and ChRAD genes in Chirita heterotricha (Gesneriaceae)

Abstract CYC‐like genes are widely conserved in controlling floral dorsoventral asymmetry (zygomorphy) through persistent expression in corresponding domains in core eudicots. To understand how CYC‐like gene expression is maintained during flower development, we selected Chirita heterotricha as a material and isolated the promoter sequences of the ChCYC1C and ChCYC1D genes, homologs of CYC, by inverse polymerase chain reaction. Further promoter analyses led to the identification of a putative cis‐regulatory element in each promoter matching the consensus DNA binding site for Antirrhinum CYC protein: GGCCCCTC at ?165 for ChCYC1C, and GGCCCCCC at ?163 for ChCYC1D. This indicates that both the ChCYC1C and ChCYC1D genes have probably evolved autoregulatory loops to sustain their expression in developing flowers. We also isolated the coding and promoter sequences of the ChRAD gene, a homolog of Antirrhinum RAD. Promoter analysis showed that the ChRAD gene promoter also contained a putative CYC‐binding site (GGCCCAC at ?134). Therefore, ChRAD is likely a direct target of the ChCYC1 genes, which is similar to Antirrhinum RAD. These results imply that the establishment of floral zygomorphy in Chirita may have been achieved by the evolution of an autoregulatory loop for CYC‐like genes, which was probably accompanied by simultaneous co‐option of the RAD‐like gene into their regulatory network.