Influenza A H5N1 clade 2.3.4 virus with a different antiviral susceptibility profile replaced clade 1 virus in humans in northern Vietnam

Background

Prior to 2007, highly pathogenic avian influenza (HPAI) H5N1 viruses isolated from poultry and humans in Vietnam were consistently reported to be clade 1 viruses, susceptible to oseltamivir but resistant to amantadine. Here we describe the re-emergence of human HPAI H5N1 virus infections in Vietnam in 2007 and the characteristics of the isolated viruses.

Methods and Findings

Respiratory specimens from patients suspected to be infected with avian influenza in 2007 were screened by influenza and H5 subtype specific polymerase chain reaction. Isolated H5N1 strains were further characterized by genome sequencing and drug susceptibility testing. Eleven poultry outbreak isolates from 2007 were included in the sequence analysis. Eight patients, all of them from northern Vietnam, were diagnosed with H5N1 in 2007 and five of them died. Phylogenetic analysis of H5N1 viruses isolated from humans and poultry in 2007 showed that clade 2.3.4 H5N1 viruses replaced clade 1 viruses in northern Vietnam. Four human H5N1 strains had eight-fold reduced in-vitro susceptibility to oseltamivir as compared to clade 1 viruses. In two poultry isolates the I117V mutation was found in the neuraminidase gene, which is associated with reduced susceptibility to oseltamivir. No mutations in the M2 gene conferring amantadine resistance were found.

Conclusion

In 2007, H5N1 clade 2.3.4 viruses replaced clade 1 viruses in northern Vietnam and were susceptible to amantadine but showed reduced susceptibility to oseltamivir. Combination antiviral therapy with oseltamivir and amantadine for human cases in Vietnam is recommended.     

Procedure for Evaluating the Effects of 2,450- Megahertz Microwaves upon Streptococcus faecalis and Saccharomyces cerevisiae

Modifications of a commercial 2,450-megahertz microwave oven were made so that 6 ml of microbial suspension could be exposed to the microwave field for various periods of time. The microorganisms were contained in the central tube of a modified Liebig condenser positioned in the approximate geometric center of the oven cavity. Kerosene at -25 C was circulated through the jacket of the condenser during microwave exposure permitting microwaves to reach the microbial suspension. Flow rates of the kerosene were varied to permit the temperature of the suspension to range from 25 to 55 C during microwave exposure. Conductive heating experiments using similar temperatures were also conducted. A thermocouple-relay system was employed to measure the suspension temperature immediately after the magnetron shutoff. Continuous application of microwaves to suspensions of 10(8) to 10(9)Streptococcus faecalis or Saccharomyces cerevisiae per ml appeared to produce no lethal effects other than those produced by heat. Respiration rates of microwave-exposed Scerevisiae were directly related to decreases in viable count produced by increased microwave exposure times.     

The relative effectiveness of 6-thioguanine and 8-azaguanine in selecting resistant mutants from two V79 Chinese hamster cells in vitro

The frequency of surviving colonies in two V79 cell lines exposed to either 6-thioguanine or 8-azaguanine was dependent on initial plating density. Different degrees of metabolic-co-operation were found to occur in the two cell lines and the loss of both spontaneous and added mutants occurred at a lower cell density when 6TG was used for selection than when 8 AZ was used in both cell lines. Both analogues were degraded on incubation in medium plus serum in the absence of added cells. Variation in serum batch had little effect on the rate of degradation or on the frequency of colonies recovered after treatment of V79 cell lines with 8AZ. The reasons for preferring 8AZ to 6TG as a selective agent are discussed.     

Transplantation of tumour with a kidney graft.

A cerebral glioma discovered by angiography and brain biopsy in a kidney donor was subsequently suspected of being a secondary tumour. By this time a biopsy of one of the transplanted kidneys had shown a clump of malignant cells in a glomerulus. Because of the psychological state of this recipient the transplant was not removed, but the recipient of the second kidney was immediately told of the danger of tumour cell transfer, and underwent nephrectomy. The patient remained well on haemodialysis; multiple sectioning of the kidney showed no signs of tumour. The transplant in the first recipient functioned well until his death, six months after operation. At necropsy undifferentiated tumour was found in the pleura, liver, pelvic peritoneum, and transplanted kidney. All cadaver donors should undergo full laparotomy after removal of the kidneys, particularly those with a high risk of cancer, and a full necropsy should also be performed shortly afterwards to exclude tumour and other unsuspected diseases. Then it is not too late to remove a transplanted kidney should a tumour be found.     

KLF8表达修饰及致瘤机制研究进展

KLF8(Krüppel-like factor 8)是Krüppel 样转录因子家族最新成员。KLF8广泛表达于皮肤、脂肪、食道等组织中。早期研究表明,KLF8蛋白具有转录抑制活性,在胚胎发育过程中起重要作用。而近些年大量的研究发现KLF8在多种肿瘤组织中异常高表达,具有转录活化因子活性,可通过对细胞增殖、迁移、侵袭、EMT、细胞干性等多项生理过程调控促进肿瘤发生发展。综述了近年来KLF8研究进展,系统阐述了KLF8表达修饰调控及致瘤机制,为全面深入了解KLF8在肿瘤中作用及后续相关研究提供参考。     

Combining nucleoside analogues to achieve recognition of oligopurine tracts by triplex-forming oligonucleotides at physiological pH

We have used DNase I footprinting to examine DNA triple helix formation at a 12 base pair oligopurine.oligopyrimidine sequence, using oligonucleotides that contain combinations of 2'-aminoethoxy-5-(3-aminoprop-1-ynyl)uridine (bis-amino-U, BAU) and 3-methyl-2-aminopyridine (MeP) in place of T and C, respectively. This combination acts cooperatively to enable high affinity triple helix formation at physiological pH. The affinity depends on the number of substitutions and their arrangement; oligonucleotides in which these analogues are evenly distributed throughout the third strand bind much better than those in which they are clustered together.     

小麦和无融合生殖披碱草杂交后代（BC2F2）的无融合生殖及胚胎发育过程中的异常现象研究

对小麦（Triticum aestivum）和无融合生殖披碱草（Elymus rectisetus）的染 色体数目为42的杂种后代（BC2F2）单株进行了RAPD检测和胚胎学研究。RAPD检测结果表明：染色体数目为42条的BC2F2单株的遗传组成与普通小麦的遗传组成十分接近,但是在部分单株中出现了披碱草的特异带。由此可以推测,经过回交和自交后小麦草的部分染色体片段已经整合进了小麦的染色体。在部分BC2F2单株胚胎学切片中发现了较高比例的（5%左右）双孢原、早发胚以及多胚囊等无融合生殖现象,直接表明了无融合生殖基因转移。由于基因整合的多样性,无融合生殖基因在有些单株中并没有充分表达,从而造成了某些单株胚胎发育的异常。     

脆性组氨酸三联体基因小干扰RNA的构建及干扰效果检测

目的:构建脆性组氨酸三联体(FHIT)基因的小干扰RNA(siRNA),并检测其对293T细胞内源性FHIT基因表达的干扰效果。方法:根据FHIT基因序列,通过生物信息学网站预测可能的siRNA,从中筛选出2条合理的FHIT-siRNA,将其克隆到siRNA表达载体pSilencer2.1-U6Hygro中,转化大肠杆菌DH5α,挑取阳性克隆进行测序鉴定;将构建成功的FHIT-siRNA重组载体转染人胚肾细胞293T,Western印迹检测siRNA对293T内源性FHIT表达的干扰效果。结果:构建了2个FHIT-siRNA重组质粒,2条siRNA都有干扰作用,其中一条的作用效果可达50%以上。结论:构建的FHIT-siRNA为进一步研究FHIT的功能提供了有利的工具。     

几种小分子化合物对P53蛋白与SV40大T抗原之间相互作用的影响

以P53蛋白与SV40大T抗原之间的相互作用作为药物分子靶标,利用酵母双杂交系统的α-半乳糖苷酶活力的测定方法,测定了两个系列的化合物对于P53蛋白与SV40大T抗原相互作用的影响。其中一个系列包括ZnCl2,MgCl2,CaCl2,和Cu-Cl2、BaCl2等二价金属盐;另一个系列是几种新疆特色植物的天然提取物,包括蒜氨酸、大蒜素、罗丹明B类物质AJ-8、黄酮、苦参碱、去氢骆驼蓬碱和骆驼蓬碱等。结果发现,ZnCl2能够特异使P53蛋白与SV40大T抗原之间的相互作用增强15％左右,而大蒜素和黄酮能够降低这两个蛋白的相互作用,降低程度分别为40％和30%,其它药物基本不影响两个蛋白的相互作用。