关键词： 脊髓灰质炎   山东   医科大学   小儿麻痹   肠道病毒感染   医学院校   菌苗   疫苗   儿童基础免疫  

摘要： ３３例脊髓灰质炎疫苗服用情况分析山东医科大学丛滋敏，马金耀，袁芳济南中心医院王述昀脊髓灰质炎减毒活疫苗是儿童基础免疫措施之一，基础免疫质量是其关键。为此，将１９８９年济南市传染病医院收治的脊髓灰质炎３３例的基础免疫情况进行了分析。患者入院后，医生详细...

关键词： Animals   Base Sequence   Capsid   Cell Line   DNA   Viral   Defective Viruses   Epitopes   Hela Cells   Humans   Mice   Molecular Sequence Data   Mutation   Neutralization Tests   Poliovirus   Receptors   Virus   Research Support   U.S. Gov't   P.H.S   Serotyping   Virus Replication  

摘要： Several mouse cell lines expressing hybrid human poliovirus receptors (hPVRs) bearing mutations in the first immunoglobulin-like domain were previously characterized for their defective binding and replication of poliovirus type 1 Mahoney (G. Bernhardt, J. Harber, A. Zibert, M. DeCrombrugghe, and E Wimmer, Virology, 203, 344-356, 1994). Here we report that these mutant hPVRs were utilized to explore differences in the binding behavior of the three serotypes of poliovirus. Type 3 polioviruses (both Sabin and the neurovirulent Leon strain) clearly bound to the hPVR mutant Q130G/GD, but were incapable of initiating infection. Also, binding at 25 degrees of poliovirus types 2 and 3 to cell lines expressing the hPVR mutants P84SYS/HFGA, L99GAE/AAAA, and D117F was greater than type 1 poliovirus. Further study of the serotype-specific interaction with mutant hPVRs was accomplished with antigenic hybrid viruses. Improved binding by antigenic hybrid viruses demonstrated that serotype-specific binding lo mutant hPVRs is, in part, determined by the amino acid sequence of neutralization antigenic sites (NAgs) and the probable conformational rearrangement of amino acids adjacent to the NAg sites. Finally, site-directed mutants of poliovirus were utilized to determine the relative contributions, to hPVR interactions, of individual amino acids with solvent accessible side chains in the viral canyon. Of the 18 viable virus mutants produced, 3 (D1226A, 11089A, and VPEK1166HPGA) expressed impaired replication phenotypes on the mutant hPVR cell lines P84SYS/HYSA and D117F. A location at the rim of the poliovirus canyon was implicated for the interaction of the amino terminal domain of the poliovirus receptor with conserved and serotype-specific viral surface amino acids. The possible involvement of elements of neutralization antigenic sites in receptor binding may explain, in part, why poliovirus exists in only three serotypes. (C) 1995 Academic Press, Inc.

关键词： protease 2A   poliovirus   p220 cleavage   vaccinia virus  

摘要： The effects of transient expression of poliovirus 2A(pro) on p220 cleavage in COS cells have been analyzed. When 2A(pro) was cloned in plasmid pTM1 and transiently expressed in COS cells, efficient cleavage of p220 occurred after infection of these cells with a recombinant vaccinia virus bearing phage T7 RNA polymerase. High numbers of COS cells were transfected with pTM1-2A, as judged by p220 cleavage, thereby allowing an analysis of the effects of poliovirus 2A(pro) on vaccinia virus gene expression. A 40-50% cleavage of p220 by transfected poliovirus 2A(pro) was observed ten hours post infection and cleavage was almost complete (80-90%) 20-25 hours post infection with vaccinia virus. Profound inhibition of vaccinia virus protein synthesis was detectable ten hours post infection and was maximal 20-25 hours post infection. This inhibition resulted from neither a blockade of transcription of vaccinia virus nor a lack of translatability of the mRNAs present in cells that synthesize poliovirus 2A(pro). Addition of ara-C inhibited the replication of vaccinia virus and allowed the continued synthesis of cellular proteins. Under these conditions, 2A(pro) is expressed and blocks cellular translation. Finally, p220 cleavage by 2A(pro) did not inhibit the translation of a mRNA encoding poliovirus protein 2C, as directed by the 5' leader sequences of encephalomiocarditis virus. Therefore, these findings show a correlation between p220 cleavage and inhibition of translation from newly made mRNAs. Our results are discussed in the light of present knowledge of p220 function, and new approaches are considered that might provide further insights into the function(s) of initiation factor eIF-4F.

关键词： protease 2A   poliovirus   p220 cleavage   vaccinia virus  

摘要： Poliovirus protease 2A(pro) has been efficiently expressed in HeLa and COS cells upon transfection with vector pTM1-2A and infection with the recombinant vaccinia virus bearing the T7 RNA polymerase. The expressed poliovirus protease localizes to the cytoplasm of the transfected cells, both in the endoplasmic reticulum and in vesicles scattered in the cytoplasm, Cleavage of p220, a component of initiation factor eIF-4F, selectively occurs from 5 h post-infection in transfected cells infected with the recombinant virus, This cleavage correlates in time with the profound inhibition observed in the synthesis of vaccinia virus proteins, A similar blockade of vesicular stomatitis virus translation takes place upon 2A(pro) expression. Finally, the synthesis of poliovirus protein 2C from a recombinant vaccinia virus that expresses this protein under the EMC untranslated leader region is not affected by the synthesis of 2A(pro). These findings lend support to the idea that translation of capped mRNAs requires the integrity of p220, while this requirement is not observed when translation of a mRNA bearing a picornavirus leader region is assayed.

关键词： 聋哑患者   听力学检查   病因分析   听力语言训练   耳毒性药物   类抗生素   预防措施   脊髓灰质炎减毒活疫苗   脑干听觉诱   氨基糖  

摘要： 听力障碍多为感音性聋,目前治疗比较棘手,因而对其病因进行分析,采取针对性预防措施非常重要。对聋哑患者进行听力学检查,早确诊,早期进行听力语言训练是非常必要的。 1 临床资料 1987年1月至1994年12月来我科就诊的聋哑患者,详细询问病史及家族史,全面查体及进行纯音测听和声导抗测试检查,年龄小不能配合者行脑干听觉诱发电位测定。资料完整的150例,男87例,女63例,

关键词： enterovirus 71   viral diversity   poliovirus   coxsackievirus A16  

摘要： Enterovirus 71 (EV71) is capable of causing paralytic disease indistinguishable from poliomyelitis due to poliovirus. To determine the relationship of EV71 to poliovirus and other enteroviruses, two strains of EV71 have been cloned and sequenced. The EV71 strains had only 46% amino acid identity with the polioviral P1 capsid region and 55% with the entire polyprotein. There were no regions of high similarity that might account for their respective ability to cause paralytic disease. The two strains, a neurovirulent isolate (EV71/7423/MS/87) and the prototype strain (EV71/BrCr), share 81% nucleotide identity and 95% amino acid identity. Sequence comparisons in the coding region between the two EV71 strains and other picornaviruses indicate that EV71, coxsackievirus A16 and coxsackievirus A2 comprise a distinct genetic group within the enteroviruses.

关键词： Animals   Antigens   Viral   Chickens   Cooperative Behavior   Guinea Pigs   Immunogenetics   Poliovirus Vaccine   Inactivated   Rats   Reference Standards   World Health Organization  

摘要： Ten laboratories studied the immunogenicity of six trivalent inactivated poliovirus vaccines in a WHO Collaborative Study. The antigenic content of all six vaccines had previously been estimated in an earlier study (Wood et al., 1995). In collaboration with the European Pharmacopoeia Commission an additional preparation, European Pharmacopoeia Biological Reference Preparation Batch 1, was also included. Laboratories were requested to use established immunogenicity assays and six used the European Pharmocoepia guinea pig/chick test while three used a rat potency test. One laboratory contributed data from both methods. Apart from one laboratory, within laboratory variation was low (less than five-fold). However, very large (greater than 100-fold) variation was seen between laboratories for ED50 results in the guinea pig/chick test. Different decisions on pass/fail outcome would have occurred for some of the samples tested. Between laboratory variation was much lower (less than fivefold) in the rat test. Expression of results as potencies relative to a standard reduced between laboratory variation for both methods, substantially so for the guinea pig/chick test. The correlation between in vivo and in vitro results was generally good with the exception of the type 3 component of one preparation. This showed that the relationship between immunogenicity and antigenicity was not necessarily predictable. There is an urgent need to revise the European Pharmocoepia immunogenicity test but it is premature, on the basis of this study, to recommend either the rat test or in vitro tests as replacements. Two candidate reference materials were both found suitable for in vivo assay of inactivated poliovirus vaccine. (C) 1995 The International Association of Biological Standardization

关键词： TRANSCRIPTION   TEMPLATE INTERACTION   SECONDARY STRUCTURE   MUTATIONAL ANALYSIS   3'-UNTRANSLATED REGION   PICORNAVIRUS  

摘要： A series of mutations were introduced in the 3'-terminal untranslated region (3'-UTR) of full-length infectious poliovirus cDNA clones, and following transfection of COS-1 cells the ability of these constructs to generate viable viral particles and/or to support viral RNA synthesis was assayed. Substitution of the 3'-UTR of poliovirus RNA with the equivalent sequences of HAV RNA abrogated viral RNA replication, whereas the introduction of extended 'foreign' sequences between the open reading frame and the 3'-UTR was well tolerated. Point mutation that either destabilized the stem-and-loop structure or altered the sequence of the loop in domain 'Y' (nomenclature as per Pilipenko et al., [Nucleic Acids Res. 20 (1992) 1739-1745]) abolished both the infectivity and viral RNA synthesis. These were not restored by compensatory mutation that reconstructed the native secondary structure of this domain, suggesting that the secondary/tertiary folding of the 3'-UTR is not the only determinant for template recognition at initiation of RNA synthesis, but rather that a specific primary sequence is indeed required.

关键词： POLIOVIRUS   ENTEROVIRUS   ALS-MOTOR NEURON DISEASE   SPINAL CORD   POLYMERASE CHAIN REACTION  

摘要： A longstanding hypothesis proposes that amyotrophic lateral sclerosis-motor neurone disease (ALS-MND) is a late consequence of subclinical poliovirus (PV) infection. In this study, RNA extracts of CNS tissue from 28 patients with ALS-MND and 7 controls were assayed by nested polymerase chain reaction (PCR) using primers to the 5'-untranslated region (UTR) of the enterovirus (EV) genome which is highly conserved between EVs including PV, echovirus and coxsackie viruses. The integrity of RNA extracted from either archival paraffin-embedded or frozen CNS tissue was assessed by detection of constitutive Ableson tyrosine kinase (ABL) mRNA by PCR. Of 63 tissue samples assayed, 81% (51/63) were ABL-positive corresponding to 78% (22/28) of the ALS-MND cases and all controls, None of the 27 ALS-MND cases (i.e, 21 ABL(+) and 6 ABL(-)) in which paraffin-embedded tissue was used nor any of the age and sex matched controls were positive for specific PV/EV RNA. Moreover, CNS tissue from 14 different locations obtained from one patient < 2 hrs after death and immediately frozen, showed no evidence of PV/EV at any site by PCR. Disease duration, degree of tissue autolysis and duration of tissue storage were all excluded as factors which may predispose to negative results. The sensitivity of the PV PCR was determined to be 40-400 copies (12.5-125 ag) of synthetic EV RNA transcripts in i mu g of cellular RNA and the assay was shown to detect all types of PV and other EVs tested. Thus it seems unlikely that a persistent PV or related EV infection is implicated in ALS-MND unless there has been alteration in the 5'-UTR of the virus genome.

关键词： poliovirus   China   eradication   PCR-RFLP   genotypes   epidemiology  

摘要： We examined the genomic variability of 26 wild-type 1 polioviruses isolated from 12 provinces in China from 1985-1993, The poliovirus genome region that encodes the N-terminal portion of the VP1 protein was amplified by the polymerase chain reaction (PCR), The PCR product of each isolate was analysed by restriction fragment length polymorphism (PCR-RFLP) and by direct sequencing. Several groups of viruses with different PCR-RFLP patterns were found to cause outbreaks in China during this period. Viruses with identical PCR-RFLP profiles were dominantly isolated in 11 provinces from 1991 to 1993 and were considered to be the major epidemic strain in China. On the other hand, in the GuangDong province, a different group was responsible for each year's outbreak, while in the XinJiang autonomous prefecture, a single strain was consistently isolated from 1990 to 1993, Understanding of the nationwide distribution of wild poliovirus is very important for interrupting its transmission during the final stage of the eradication program in China.