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1995 Vol.10(4)

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Review

The study on adaptation of Japanese encephalitits virus in Veto cells

DAN Hui-Ying, DING Zhi-Fen, CHANG Zhen-Yan

1995, 10(4): 273

3 strain of Japanese Encephalitis(JE)virus was adapted in Vero cells for 27 passages;Nosignlficant changes were found on virus titre and CPE after adaptation,but the immunogenicitywas getting weaker after 10 passages.The immunogenicity weakened could be recovered partlallyby repassag ing the V ero-cells adapted virus in mouse brain,Less than 10 passages of ad aptedviru s growing in Vero cells could be used as seed virus for prod uction of JE vaccine instead ofvirus prepared from mouse brain.U sing the ad apted virus for vaccine production could be easierto avoid the contamination of extraneous agents com pared with using the virus from mouse brain.

Distribution of virus antigen and pathologic changes in the tissues of rabbits infected with EHFV

LIN Yu-Lin, BANG Gong-Mei, GAO You-Xin, ZHANG Wei-Ying, CHEN Hong-Bin, XIANG Jin-Min

1995, 10(4): 278

Rabbits were inoculated with EHFV-H114 strain,the distribution of virus antigen in thetissues of rabbits was studied by immunoflourescent technique and double bridge peroxidase(PAP)method.Meantinle,pathologic change in the tissues embedded in paraffin was observedby H-E staining.The results showed that the viral antigen was detected in every tissue 7-15days after infection by IFA,PAP staining,the positive cells were mainly the endothellcal cells ofcapillary,small blood vessele in every tissue detected,the results ind icated that the endothelialcells of capillary,small blood are EHFV target cellg,Also,the EHFV antigens were detectedfrom glond cells of both ovary and testis of the rabbits,it provides EHFV vertical tranmissionwith the data of etiology.The specific antigens were also found in epithelial cells of bronchus ofthe rabbits,it suggests that the epithelial cell be susceptible to EHFV,and there be Dossibility ofEHP aerosol spread.Prom the observation of the pathologic change in positive tissues,

Experimental Studises Oil Viral Hepatitis Receptor——targeted Drug L——HSA-AraAMP

ZHANG Ling-Xia, WANG Hui-Fen, LI Ke, ZHONG Bo-Hua, LIU Hong, PAN Yong

1995, 10(4): 283

Lactosaminated Human Serum Albumin(L-HSA)was,used as a carrier and coupled to Ara-binosied adenine monophospate(AraAMP).The distribution of 125I-labelled L-HSA-AraAMPwere higher in liver than in other organs.Radioactivities of the drug could still be measured atthe 7th dav after injection.The efficacy of targeted drug was determined by analyzing dynamic changes of HBsAg andHBeAg in 2.2.15 cell culture medium by RIA and changes in the replicative intracellular HBVDN A content by Southern blot.The results showed that the targeted drug had significantly in-hibitory ratio of HBsAg and HBeAg which was similar to AraAMP(inhibitory ratfo of HBsAg50%,63%;HBeAg 62.6%,67.2%,respectively).Southern blot test showed the replicativeintermedlates rcDNA and ssDNA were inhibited by AraAMP.The targeted drug had parallel an-ti-H BV DN A reDlicative effectiveness com pared with AraAMP.The values of Si of drugs toHBsAg and HBeAg were detected.

Selection, synthesis and application of hepatitis E virus peptides

QI Zhong-Tian, PAN Wei, CUI Da-Fu, YU Chao, CUI Heng-Ran

1995, 10(4): 290

Accordingto our computer analysis of hydrophobicity and prediction of secondary structuresfor the full-length putative proteins encoded by ORF 1(open reading frame-1),ORF2 and ORF3ofhepatitis E virus(HEV),we selected amino acid regions with tiydrophilicity,β-turn,and β-sheet,and synthesized seven peptities of possible epitope-containing regions of the polypeptide en-coded by all three ORFs of HEV genomic RNA by Merrifield s method of solid-phase,The syn-thetic peptides were screened and identified by solli-phase enzyme-linked immunosorbent assay(ELISA).Three of the peptides(EH174 from ORF1,EH286 from ORF2 and EH362 from ORF3)showed antigenic activity and possible application for the development of anti-HEV test k its(thepeptide-based EL ISA).The laboratory experunents and clinical trials indicated that the kits,us-ing a set of three synthetic HEV pepties as coating antigens,were of high specificity and exhibit-ed good reproducibility,Additionally,our results also demonstrated good agreement with clinical find i

Inhibitory Effect of TNF-a on Human Common Respiratory Tract Viruses in vitro

YAO Wang, WU Xiao-Ling, ZHOU Feng, ZHOU Yao-Xi

1995, 10(4): 298

This paper reports the protective effect of human rTNF-α and Human IFN-α on HEL andNEP-2 cell cultures infected by viruses in vitro,That is,before virus challenge,the treatment ofcell cultures in comparison with TNF-αand IFN-α may result in the inhibition of virus inducedcytopattiy.The challenge viruses included 5 strains of different tyDe of adenovirus,1 strain ofNSV-1,1 strain of rhinovirus,1 strain of Sendai virus,and 1 strain of VSVS,The antiviral ac-tivity of TNF-αand LFN-αare specific,and with wide spectrum.The specific antiviral activity ofboth TNF-αand IPN-α is confirmed by antibody neutralization. role of TNF-α inhibition can beneutralized completely by monoclonal IFN-β antibody and partly by monoclonal IFN-α antibody.The mechanism of antiviral effect of TNF-αis discussed. Aecording to the experiments ofneutralization test the TNF-α antiviral actlvity may be cumpletely neutralized by monoclonal anti-body againist IFN-β.It is strongly suggested that the antiviral effect of TNF-α appear to be me-diated

PCR amplifying, cloning and sequencing of E7 gene of human papillomavirus type 16

SU Ying-Bin, ZHAO Wen-Xian, WU Xin-Xing, DAI Tian-Li, DING Xiao-Hua

1995, 10(4): 303

Rabbits were inoculated with EHFV-H114 strain,the distribution of virus antigen in thetissues of rabbits was studied by immunoflourescent technique and double bridge peroxidase(PAP)method.Meantinle,pathologic change in the tissues embedded in paraffin was observedby H-E staining.The results showed that the viral antigen was detected in every tissue 7-15days after infection by IFA,PAP staining,the positive cells were mainly the endothellcal cells ofcapillary,small blood vessele in every tissue detected,the results ind icated that the endothelialcells of capillary,small blood are EHFV target cellg,Also,the EHFV antigens were detectedfrom glond cells of both ovary and testis of the rabbits,it provides EHFV vertical tranmissionwith the data of etiology.The specific antigens were also found in epithelial cells of bronchus ofthe rabbits,it suggests that the epithelial cell be susceptible to EHFV,and there be Dossibility ofEHP aerosol spread.Prom the observation of the pathologic change in positive tissues,

Cloning and sequencing of pSTY28 DNA fragment of human herpesvirus-6

JIAN Li-Sheng, SHAN Xi Hong Yi

1995, 10(4): 309

This report confirms the sequence of pSTy 28 fragment of HHV-6.We jave determined thecomplete nucleotide sequence,The sequence data have revealed open reading frame(ORF)of 2414 nucleotides encoding a ribonucleotide reductase (RiR) protein of 805 amino acids and 1100nucleotides encoding a P41 protein,To examine the relatedness of RiR protein with those of otherherpesvirus,we compared their amino acid Sequence in pairwise combination,Interestingly,RiRORF of HHV-6 has shown to have the highest amino acid humology with the RiR of HCMV.The optimized score reaches 475.Biochemical analvsis of RiR has ind icated the active site ofthioredoxin ontaining two cysteins separated by two amino acids.This sequence was found onlyonce in the E.coli,HSV-2,EBV and mouse RiR protein,but we did not find it in the HHV-6and HCMV RiR protein.Those data support Esftathious view that HHV-6 belongs to betaherpes viruses.

High expression of HCV core antigen and the study on the recombinant protein antigenicity

ZHANG Shu-Min, QI Zi-Bai, LIU Ke, LI He-Min

1995, 10(4): 317

Part of HCV core gene fragment is expressed in E.coli.The recombinant protein C27 is fu-sion protein with GST.The specific protein iS about 30%of total bacterial protein,After purifi-cation,the purity of the recombinant protein is more than 95%.We compared C27 antigen withC11 core antigen by EL ISA using the second generation panel sera of HCV EIA d iagnostic kit.The result showed that the two antigens had the same rate of sensitivity and specificity.So,weconcluded that the expressed gene fragment may be an important region in HCV core gene,Thefusion protein may also play an important role in increasing the antigenicity of the recombinant protem.

Cloning of parvovirus H-1 NS partial gene by PCR

HUANG Qing-Shan, HUANG Jian, LUO Jie-Yu, WANG Shun-De, LIU Wei-Ping

1995, 10(4): 323

Two modified primers,△P3 and △P4,were designed and,for each of them,two mutatedbases were included.A fter PCR amplification, the DNA fragment contains a restriction endonu-clease recognition site,BamHⅠ or Hind Ⅲ at its ends,Cloning of the parvovirus H-1 NS-1 partialgene was conducted by double digestion uf the aniplified DNA fragement with endonucleases,andits presence in the inserted fragement was confirmed by DNA sequence analysis.The techniquedescribed here provides the basis for the quality control of H-1 propagation,and for the studies ofoncosuppressive action of H-1 NS-1 protein and of transcription and expression of NS-1 gene incancer cells and normal tissues.

A report on the research of virus populations on pepper ( Capsicum annuurn L. ) and their distributions in six provinces or cities of China

YANG Yong-Lin, YAN Chu-Zhen, TIAN Ru-Yan, FENG Lan-Xiang, LIANG Xun-Sheng, WANG Zhi-Yuan

1995, 10(4): 332

From 1983 to 1991,our collaborating research group of the project(one of the main stateproject of scientific and technical research during the period of The Sixth-Seventh Five Yearplan)collected and studied 3618 portions of san1ples of hot or sweet pepper virus disease in Bei-jing,Tianjin, Nianjing,Liaonin and Xinjiang.Eight kinds of virus(TMV CMV PVX PVY TEVAMV BBWV and TRV)were isolated from the samples by three routine methods of identifica-tion(biological,seroloical and electron microscopical method).In the pepper,the biological,seto-logical and electron microscopical characters of these viruses and symptoms they caused on hot orsweet peppers were described,the alternating laws between decline and increase of all kinds ofvirus in various both region and seasons for seven years were analysed,the kinds of dominantvirus on pepper in each region was stud ied clearly,The results would provide scientific founda-tions for both antiviral breeding and making of the intergrated control plan.

Identification of Virus Diseases of Tobacco in Hellongjiang Province and Changchun City

WEI Pei-Wen, ZHANG Ming-Hou, LV Wen-Qing

1995, 10(4): 340

ne hundred and twenty nine samples of virus infected tobacco leaves were collected from 11counties/cities of Heilongjiang Province and Changchun City during early June to late August,1991-1993.Viruses were identified by differential hosts,serological assays(ELISA or DIBA)and electronmicroscopy of ISEM.TMV,PVY,CMV,TRV,TSWV and TMV-Y were foundisolately or complicatedly.The most predominate virus was TMV,and the next one was PVY,their rates of occurence were 53.5%and 40.3%,respectively

The Priliminary Study on Narcissus Infected with PVYG roup Viruses by Degenerate PrimerPCR Techniques

CHEN Zhi-Nan

1995, 10(4): 346

More than four PVY group viruses were found to infect Narcissus by the techniques of de-generate primer reverse transcription(RT)-polymerase chain reaction(PCR)and restriction frag-ment length polymorphism(RFLP),they may be TBV,NYSV,NSSV and OnYDV etc.

Rapid Detection of Proviral DNA of Bovine Leukaemia Virus Using Nested PCR

WANG Han-Zhong

1995, 10(4): 351

This paper reported that proviral DNA of bovine leukaemia virus(BLV)was detected bynested PCR.External and internal primers were selected from gp51 gene in BLV genome,Theproducts of the nested PCR were identified by 2%a8arose 8el electrophoresis and were com-flrmed by hybridlzation with a biotinylated oligonucleotide probe,The results of the nested PCRshow that DNA forms of the provirus were detected from two host cells.

Isolation,Putjfication of Pem em chinesis Parvovirus and Analysis of its Nueleic Acid and Protein

XIAO Lian-Chun, SHI Zheng-Li, GAO Wei, ZHANG Li-Ren, CHEN Dai-Hua

1995, 10(4): 356

Small spherical virus with diameter of ca,20 nm was isolated from infected shrimps.Thevirions were icosahedron and naked. The infection exDeriment showed that the mortality was upto 66%.The viral nucleic acid was ssDNA after digested with DNase,RNase and Sl nuclease,Analysis of viral polypeptides by SDS-PAGE revealed that the virion had four structural polypep-tides with MW 86 kD,79kD,70 kD and 25.5 kD.These resul ts were in comformity with Par-voviridae,thus it was proposed that the virus be named Penaeus chinesis Parvovirus.

A Broad spectrum Antiphage Strain Constituted with Protoplast Fusion Technology

QIU Juan-Ping, ZHOU Ning-Yi

1995, 10(4): 362

Antiphage mutants selected by 15 phage strains have resistant only to its relevant phage,50it is narrow-spectrum antiphage strain.The fusant ZG88 is obtained with protoplast fusion be-tween Corynebacterium pekinense 7338 and Corynebacterium crenatum Ts-Li.It s serologic reac-tion and phage attach test show that it has changed so much in cell surface structures as to losethe position of adsorption.So ZG 88 is a broad spectrum and steady antiphage strain.

Prelim inary Identification Cucumber Mosaic Virus and Serological Detection of Infecting Chrysanthemum

ZHANG Hai-Bao, ZHU Xi-Ru, ZHANG Yun-Kai

1995, 10(4): 367

The virus isolate S_1 was isolated from chrysanthemum plants with the symtoms of viral dis-ease,The host range of isolate S_1 was wide.It could infect 11 species plants aniong tested 6 fami-lies by sap-inoculation, The isulate S_1 was also transmitted in non-persistant manner by Myzuspersicae,The rate of transmission was 86.7%(13/15).The vlrus particle of isolate S_1 wassphere stiape, ItS diameter was about 28-30nm,The serological test showed that isolate S_1 re-acted positively with the antiserum against CMV but negatively with ToAV antiserum.On thebasis of these characteristics,isolate S1 was identified as a member of cucumber mosaic virus(CMV),The rabbit antiserum against CMV(isolate S1)was prepared with purified isolate S1.The highest titer of the antiserum was 1:2400 tested by ELISA.All 158 virus disease samples of field chrysanthemum plants were detected with the prepared antiserum by indirect-ELISA,inwhich 19.0% amples showed positive reaction.