Protective efficacy in chickens, geese and ducks of an H5N1-inactivated vaccine developed by reverse genetics

We generated a high-growth H5N1/PR8 virus by plasmid-based reverse genetics. The virulence associated multiple basic amino acids of the HA gene were removed, and the resulting virus is attenuated for chickens and chicken eggs. A formalin-inactivated oil-emulsion vaccine was prepared from this virus. When SPF chickens were inoculated with 0.3 ml of the vaccine, the hemagglutinin-inhibition (HI) antibody became detectable at 1 week post-vaccination (p.v.) and reached a peak of 10log2 at 6 weeks p.v. then slowly declined to 4log2 at 43 weeks p.v. Challenge studies performed at 2, 3 and 43 weeks p.v. indicated that all of the chickens were completely protected from disease signs and death. Ducks and geese were completely protected from highly pathogenic H5N1 virus challenge 3 weeks p.v. The duration of protective immunity in ducks and geese was investigated by detecting the HI antibody of the field vaccinated birds, and the results indicated that 3 doses of the vaccine inoculation in geese could induce a 34 weeks protection, while 2 doses induced more than 52 weeks protection in ducks. We first reported that an oil-emulsion inactivated vaccine derived from a high-growth H5N1 vaccine induced approximately 10 months of protective immunity in chickens and demonstrated that the oil-emulsion inactivated avian influenza vaccine is immunogenic for geese and ducks. These results provide useful information for the application of vaccines to the control of H5N1 avian influenza in poultry, including chickens and domestic waterfowl.     

The role of dorsal respiratory group neurons studied with cross-correlation in the decerebrate rat

We examined the role of dorsal respiratory group (DRG) inspiratory neurons as transmitters of respiratory drive to phrenic and intercostal motoneurons and as relays of afferent information to ventral respiratory group (VRG) bulbospinal, inspiratory neurons. Attempts to antidromically activate 76 DRG neurons from the spinal cord at the C7 segment resulted in only 4 (5.3%) successes (3 contralateral, 1 ipsilateral). Cross-correlating DRG neuron discharge with that of the ipsilateral (56) and contralateral (20) phrenic nerve detected common activation peaks in 2 and 3 cases respectively, with no evidence for monosynaptic connections. Cross-correlating DRG neuron discharge with that of bulbospinal, inspiratory VRG neurons found some evidence for interaction. Peaks in 7 of 73 (10%) cross-correlation histograms were attributed to a monosynaptic excitation of DRG neurons by VRG neurons, although a common activation cannot be ruled out; troughs, some with an accompanying peak, in 9 (12.3%) histograms were interpreted as a combined excitation of the DRG neuron and delayed inhibition of the VRG neuron. In addition, 2 cross-correlation histograms showed peaks with latencies and half-amplitude widths consistent with a disynaptic excitation of a DRG neuron by a bulbospinal inspiratory VRG neuron. Cross-correlating the discharge of 57 pairs of DRG inspiratory neurons (6 contralateral) detected common activation peaks in 7 (12.3%) cases (none contralateral) and one case interpreted as evidence for a disynaptic excitation. These findings suggest that the role of the DRG inspiratory neurons in rats differs from that in cats, primarily because they do not act to transmit respiratory rhythmic drive directly to phrenic and intercostal motoneurons. The results offer some support for an excitation of DRG neurons by VRG inspiratory neurons, but no support for a role of DRG inspiratory neurons as mediators of afferent information transfer to VRG bulbospinal inspiratory neurons.     

长期慢性脑外伤大鼠脑线粒体功能的变化

研究长期慢性轻度脑外伤对大鼠脑线粒体功能的影响。大鼠连续1、5、10、15、20、25、30d轻度闭合性颅脑撞击后分离脑线粒体,测定线粒体肿胀度、膜流动性、膜磷脂含量、呼吸功能、线粒体呼吸酶、超氧化物歧化酶(SOD)、丙二醛(MDA)和Ca2 等指标以显示线粒体功能、抗氧化能力的变化。结果显示,第15、20、25、30d大鼠脑线粒体明显肿胀,膜磷脂降解,膜流动性下降,呼吸功能衰减,呼吸酶、SOD活性降低,Ca2 、MDA含量升高。由此认为,经常性头部撞击可造成大鼠脑线粒体功能受损,其机制可能与脑线粒体膜损伤后继发的自由基生成增加、脑线粒体能量代谢障碍有关。     

A polymorphism in the angiotensin 1-converting enzyme gene is associated with damage to cerebral cortical white matter in Alzheimer's disease

The impact of the insertion (I)/deletion (D) (I/D) polymorphism in the angiotensin 1-converting enzyme (ACE) gene on the extent of white matter myelin loss (ML) was investigated in four regions of the cerebral cortex in an autopsy-confirmed series of 93 patients with Alzheimer's disease (AD). The possible influence of APO E epsilon4 allele acting in concert with ACE D allele was assessed. The extent of ML did not differ between D/D, I/D and I/I genotype groups when data from all four brain regions were combined. However, separate analysis showed that the frontal and temporal cortex tended to be affected more severely by ML in patients with D/D genotype compared to those with I/D and I/I genotypes. Stratification according to APO E epsilon4 allele revealed a greater overall ML in patients bearing at least one copy of ACE D allele and one APO E epsilon4 allele, especially in individuals homozygous for both. The APO E epsilon4 allele may therefore act synergistically in patients with AD (and other subjects) bearing ACE D/D genotype to increase the risk of ML, perhaps through transient ischaemic episodes consequent upon poor cardiac output associated with coronary atherosclerosis in patients with the APO E epsilon4 allele.     

脾LAK细胞培养上清中BLT酯酶活性的动态观察

脾ＬＡＫ细胞培养上清有前后两个杀伤活性高峰，以第１１ｄ为界，第一个活性高峰与上清中ＢＬＴ酯酶活性呈正相关。含血清和无血清的培养体系产生ＢＬＴ酯酶的机理可能不同。在有高活性ｒＩＬ－２时，ＢＬＴ酯酶的分泌与无血清培养液中有无２－ＭＥ相关。     

主要常见病甲襞微循环改变重复性的研究

在目前不能直接观测病人内部器官微循环改变的条件下，应用模糊聚类的理论和方法，对主要常见病有意义的的甲襞微循环改变进行聚类，同时参考国内外相关文献的结果，提出了主要常见病有意义的甲襞微循环改变群。在改变群中任意提出3、4、5、6、7、8项改变指标，形成改变组合，一种组合重复出现二次以上的重复率，随组合改变指标数的增多而减少（70．36％～4．36％）。将改变群分成重要指标和参考指标二类，从每类中提出三项形成组合，重复率仅为0．34％。说明组合方式的影响十分巨大。经过反复探索、大量实践，有可能找出对疾病的符合率较高、漏诊率较低、误诊率较小的最佳组合方式。     

综合ICU中低年资护士培训方法探讨

目的探讨为综合ICU培养合格的护理人员的方法。方法根据ICU护士应具备的素质及低年资护士的特点,采用定期集中培训、一对一带教、参加护理查房等方法,对基础理论和基本技能进行强化,重点加强了ICU专科护理知识地训练。结果通过客观考核和主观考核,效果良好。结论经过为期12个月两个时段较规范的岗位培训,低年资护士能较好的适应ICU工作。     

Extracellular matrix activity and caveolae events contribute to cell surface receptor activation that leads to MAP kinase activation in response to UV irradiation in cultured human keratinocytes

Activation of cell surface components has been implicated in the activation of downstream signaling cascade in response to UV irradiation, and yet the identity and the interaction of those components have been scantly documented. Accumulating evidence indicates that caveolae encapsulating caveolins is the location for those interactions. We found in cultured human keratinocytes that UV irradiation induced both caveolin-1 and EGFR phosphorylation. Filipin, a caveolae disruptive agent, inhibited UV-induced caveolin-1 activation. Na+-K+-ATPase catalyzes active transport of Na+ and K+ across plasma membrane of mammalian cells, inactivation of which has recently been shown to be involved in the activation of signal transduction pathways including MAP kinase cascade. We found in this study that UV inactivated Na+-K+-ATPase in time-dependent manner, Na+-K+-ATPase activity started to decrease 5 min post UV irradiation and reduced to 60% of its original activity within 1 h. Pretreatment with Flipin and MMP inhibitor recovered Na+-K+-ATPase activity lost by UV irradiation. ECIS analysis indicated that both EGF treatment and UV irradiation increased membrane electric activity which was inhibited by MMP inhibitor and Filipin. Further study showed that pretreatment of human keratinocytes with MMP inhibitor or Filipin inhibited UV-induced phosphorylation of p38 and JNK, which was however not observed in LnCap cells, a prostate cancer cell line lacking caveolin-1. UV irradiation also induced ectodomain shedding of HB-EGF in a time-dependent manner in keratinocytes. Collectively, we conclude that UV-induced MAP kinase activation is mediated by cell surface receptor activation due to the matrix activity and membrane caveolae function and inactivation of Na+-K+-ATPase.     

The interaction between native serum albumin and hepatitis B virus

Summary Purified hepatitis B virus particles were obtained from HBeAg positive serum by sucrose gradient ultracentrifugation and sephadex G-200 gel filtration. These virions formed a precipitation line in counterimmune electrophoresis with anti-albumin antibody, but the reaction could be inhibited by anti-HBs. After two months at 4°C, another precipitating line was formed under the same condition which could not be inhibited by anti-HBs and was, thus, due to free albumin. When that sample was incubated at 37°C overnight, the line of free albumine disappeared. The virion bound albumin was monomeric in non-denaturing gel electrophoresis. These results suggest that a reversible binding between virion and albumin may occur in vivo and does not require chemical modification or cross-linking.