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1996 Vol.11(1)

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Review

The advances in PLRV genome research

ZHANG He-Ling

1996, 11(1): 1

The molecular mechanism of HIV-1 infection in the inital step

XIANG Bing-Yi

1996, 11(1): 9

Hepatitis E virus and hepatitis C virus infection in population of blood donor

LI Fang-He, YANG Dong-Liang, ZHAO Xi-Ping, YU Zhi-Qun, WANG You-Kun, HAO Lian-Jie

1996, 11(1): 17

In this study,we detected anti-HEV and anti-HCV in a group of blood donors in Wuhan areabv enzyme linked immunosorbent assay.The anti-HEV and anti-HCV positive rates were 5.74%and 9.35%,respectively,In plasma donor group,the people who had a elevanted ALT historywere 14.04% for anti-HEV and 14.18% for anti-HCV,and all of them were higher than thosefor the people who had no ALT elevanted history(4.12%and 6.45%,all P<0.01).The posi-tive rates of HBV makers between anti-HEV positive gruup and anti-HEV negative group had nosignificant difference.

Hemorrhagic fever with renal syndrome (HFRS) virus infection in the renal tissues of patients with HFRS studied by in situ hybridization and immunohistochemistry

YANG Shou-Jing, LIU Yan-Fang, HU Ti-Qi, LI Ming-Sheng, HU Ming-Hua

1996, 11(1): 21

emorrhagic fever with renal syndrome(HFRS)virus RNA and antigens were simultane-ously detected by in situ hybridization and immunohistochemical techniques in the autopsy renaltissues of the patients with HFRS.In 36 cases collected separately from Shaanxi,Shenyang,Shanghai and Jiangxi HFRS endemic foci,34 cases were subjected to virus RNA detection and 28cases were positive,34 cases were subjected to viral antigen detection and 30 cases were positive.Virus RNA and antigens were mainly localized in the cytoplasm,In the cases from Xi'an andShenyang,they were mainly distributed in the renal tubular cells.In the cases from Shanghai,Jiangxi and Guangzhou.virus RNA was mainly distributed in the renal interstitial vasculaturesand endothelial cells,occasionally in glomerular tufts and tubular cells,while viral antigens weredistributed in vasculatureb and renal tubular cells,and in Guangzhou cases in smooth muscles ofblood vessels。 Moreover,3 cases from Xi′an and 9 cases from Shanghai also displayed cytoplasmicinclusio

Detection of HCV RNA and its NS3 antigen in human primary intrahepato-cholangiocarcinoma tissues

WANG Chun-Jie, WANG Wen-Liang, WANG Bo-Yun

1996, 11(1): 29

CV RNA and its NS3 region C33c antigen were detected by in situ hybridization withdigoxgenin-11-dUTP labeled HCV 5'non coding region cDNA probe and immunohistochemicalmethods with monoclonal antibody against recombinant HCV C33c protein in 35 cases of primaryintrahepatcr-cholangiocarcinoma(PIC) tissues.The positive rate of HCV RNA in PIC was 83%(29/35) and the HCV RNA was localized in the cytoplasm and/or nucleus of cancerous cell andhepatocytes.In one case,HCV RNA was also detected in the nuclus of lymphocyte and sinusoidalcell/or kupffer′S cells.The positive rate of HCV NS3 region C33c antigen was 89%(31/35)inPIC,the positive reaction was found in the cytoplasm of cancerous cell and hepatocytes,the posi-tive cells distributed in local and diffusely.HBxAg was also detected in 77%(37/45)of the pa-tients studied.Our results suggest that HCV infection has closely relationship with the develop-ment of PIC except HBV infection.HCV infection may play an important role in the carcinogene-sis of PIC.

Biological properties of a new strain of recombinant vaccinia virus expressing rabies virus glycopro- tein

WU Lei, SHU Jia-Hong

1996, 11(1): 34

the recombinant vaccinia virus(VVM11KRG strain)contains rabies virus glycoprotein gene(RVGG) which was inserted into the Hind ⅢM fragment of vaccinia virus(V.V)genome.TheRVGG was promoted by pll promoter of V.V.The VVM11KRG does not contain lac gene.Thestudy showed that its titer is higher than that of another recombinant vaccinia virus(VVTK11KRG strain)in chicken embryo cell and CV-1 cell.Its titer reaches the highest point inchicken embryo cell in the third day and in CV-1 cell in the second day after inoculation Its sta-bility is no difference comparing with parent strain at different temperatures.The virulence ofVVM11KRG is lower than that of vaccinia virus.Western blot and IFA proved that rabies virusG protein was expressed.Southern blot showed that the RVGG was inserted into Hind Ⅲ Mfragment of vaccinia virus.The fragment between glycoprotein gene and promoter was cloned toplasmid pGEM3 zf(-).The DNA was sequenced by Pharmacia LKB DNA sequencer.Thus weobtained a clear DNA reference data for the usage of

Effect of drugs on Ca2 + influx and CVB3-RNA replication in cultured rat heart cells infected with CVB3

GUO Qi, PENG Tian-Qiang, YANG Yang-Zhen, GU Quan-Bao, ZHAO Jian-Xing

1996, 11(1): 40

he effect of Verapamil(Ver),Astragalus membranaceus(AM),Dexamethasone(Dex)on Ca ̄(2+) influx across the myocardial plasma membrane and coxsackie virus B3(CVB3)-RNA replica-tion in cultured neonatal rat heart cells infected with CVB3 was investigated.It was found thatthe Ca ̄(2+) influx could be decreased significantly (p< 0.01)by Ver,AM,Dex after infection for48h.In addtion,the aniounts of CVB3-RNA in cultured heart cells infected with CVB3、and treat-ed with AM for 48h simultaneously were significantly decreased as compared with the heart cellsinfected with CVB3 only(p< 0.001) and it was significantly higher in myocytes infected withCVB3 and treated with Ver than in myocytes only infected with CVB,(p< 0.01).These resultsdemonstrated that the above three drugs could exert the effect of decreasing the secondary dam-ages caused by Ca ̄(2+)influx,but their effects on replication of CVB3-RNA in myocardium werequite different:AM inhibited replicantion of CVB3+RNA.

Study on biologic activity for membrane and enzyme of norma} bone marrow ceils infected with epidemic hemorrhagic fever virus

ZENG Lian-Lan, LUO Duan-De, CAI Shu-Qing, LIU Wei, YANG Yu-Zhen, GUO Jin-Song

1996, 11(1): 45

sing DPH fluorescence probe,the membrane of normal bone marrow cells infected withepidemic hemorrhagic fever virus(EHFV)was labeled.The membrane lipid fluidity could becounted according to change of value of fluorescence polarization.At the same time,using cyto-histochemical staining quantitative method,the cellular ACP and SDH were detected.The resultsshowed:(1)The Lymphocyte,monocyte and granulocyte were observed dynamically after cul-tured 1,6,24,72 hours.The membrane lipid fluidity of infected cells appeared obviously low-There were significant differences statistically(p<0.05-0.01)(2)By cytohisto-chemical stain-ing and microspectrophotometer quantitating,before cultured 72 hours,ACP and SDH in infectedcells raised.However,after 120 hours,their values were decreased Also,there were significant(P<0.05).It could be related with virus invasion and damage degree of cellular function.

The application of PCR-ELISA method for detecting hepatitis C virus in serum

XU Xu-Shi, LIU Run-Zhong, LUO Yu-Ping, LI Si-Guang, ZHAO Gui-Jun

1996, 11(1): 49

Genome of the characters for the viral multiplication and cytopathic changes in Bs484 cells afterinfection with Buzura suppressaria nuclear polyhedrosis viruses(BsNPV)were reported in thispaper,The results were as follow:1.The multiplication of progeny viruses showed differentchangeable forms in the different times postinfection and the amounts of viral multiplication afterinfection with BsNPV were linearly relating to the viral infective concentration in certain range.2.The extracellular viruses in supernatant from culture were examined at 12 hours postinfection.3.The best fitted developing temperature of BsNPV in Bs484 cells was 24~26℃with regards tothe viral multiplication in vitro.4.The distinct charactistics of cytopathic changes after infectionwith BsNPV mainly included a amounts of polyhedra in nucleus,gathered infected cells and a in-flated cell with big nucleus.In addition,the two types of infected cells were observed in experi-men:a type of nucleus contained many polyhedra,another only had few polyhedr

Studies on Cyt0path0l0gical Ultrastructure of Three Different Disease Resistance Tuber Mustard Varieties Infected with Turnip Mosaic Virus

XU Jun-Huan, LI De-Bao, SHENG Fang-Jing, FANG Yue-Xian

1996, 11(1): 61

Ultrathin sections of leaf tissues of three tuber mustard varieties infected with Turnip MosaicVirus were examined in the electron microscope.The results revealed that the aggregates of fila-mentous virus particles,which always distributed near vacuole membrane and cytoplast in meso-phyll cells,were seldom found.The inclusions including pinwheels(or bundles)laminated ag-gregates and circles(or tubers)were appeared in mesophyll,epidermal and xylem parenchymascells of all diseased leaves,while the contents and the proportions of three types inclusions weredifferent in different disease resistant tuber mustard varieties,The contents of inclusions in meso-phyll cells of the sensitive tuber mustard(90-149)were more than those in resistant tuber mus-tard(90-139).Meantime the sensitive had high proportions of circles or tubers and low propor-tions of pinweels or bundles,while contrasted to the resistant.The contents and the proportionsof inclusions in the medial disease resistant variety(90-146)were bewteen above two

Influence of some antiphytoviral substances on the polymerization( in vitro) of coat protein of to: bacco mosaic virus (TMV)

JIANG Shan, GUO Xue-Liu, HAN Xi-Lai

1996, 11(1): 69

he influence of six antiphytoviral substances,including three base analogues and threemembrane lipid analogues,on the polymerization of coat protein of tobacco mosaic virus(TMV)was studied under different temperatures in vitro.The results demonstrated that all of the testedantiphytoviral substances can restrain the polymerization of TMV coat protein,The base ana-logues,including DHT,DH-DHT and Virazole,showed suppression to the polymerization processmuch more than lipid analogues, NS-83 and soybean lecithin.All the inhibition to the polymer-ization of TMV coat protein were taken place above 22℃except E-30′s.The E-30 have remark-ably inhibited the TMV coat protein polymerization.These actions suggested that all of tested an-tiphytoviral substances would be contributed to pressure the process of virion assembly.

The application of ribonuclease protection assay in the identification of strains of cucumber mosaic virus

XIE Xiang-Ming, YU Jia-Lin, LIU Yi

1996, 11(1): 69

he cDNA sequences spanning nt 1209 to nt 1626 of Fny-CMV RNA2(a subgroupⅠstrain)and nt 2002 to nt 2433 of Ls-CMV RNA2(a subgroup Ⅱ strain)were used for probe preparation.The 32P-labeled negative strand RNAs prepared by transcription in vitro were hybridized with pu-rified RNAs of two Chinese CMV isolates from pepper and tomato,respectively.The resultshows that the two Chineses isolates share high identities of sequence to the Fny-CMV,thereforethe isolates belong to the CMV subgroupⅠ strain.A transcription probe complementary to thefragment flanking nt 1657 to nt 2125 of K-CMV RNA2(a subgroupⅠstrain originated from Chi-na) was hybridized with the purified RNAs of the two Chinese CMV isolates as well.The rela-tionship between the K-CMV strain and the two Chinese isolates was compared. The applicationof ribonuclease protection assay was discussed in the identification of CMV strains.

The concentration and purification of rabies virus suspension using ultrafiltration and column chro-matography

GU Ming, SHAO Yi-Bin, GU Qi, ZENG Rong-Fang

1996, 11(1): 81

The active Rabies virus suspension was cultured in microcarrier culture of five litres bioreac-tor.After virus suspension was havested from microcarrier culture,the prdcess comprised clarifi-cation by centrifugation and filtration,concentration by ultrafiltration,purification by gel filtra-tion.The virus was inactivated with 1:4000β-propiolactone at 4℃for 24 hours.The results indicated that infective titer of the rabies virus was not declinged clearly,thatresidual calf serum protein,residual DNA content and antigenic values(NIH test)in all purifiedvirus samples were conformed to the WHO standards.The research described in this paper indicated that isolating,purifying method of active ra-bies virus was effective, It provided some basic information which could be useful in further forthe largescale production of inactivated rabies vaccines.

AA-ELISA for Detection of Barley Resistance to BYDV

LIU Chang-Hong

1996, 11(1): 84

wo viruliferous Rhopalosiphum padi(BYDV)can be detected by indirect enzyme-linkedimmunosorbent assaay(ELISA).Based on that result,a Aphid Acquisition-Enzyme-linked im-munosorbent assay(AA-ELISA)has been established for identification of barley resistance toBYDV.The OD value of AA-ELSA showed that the barley variety 877-16 carrying YD2 genewas significantly different from that of susceptible variety Manley.No significant difference wasfound between 877-16 and Manley when they were detected directly by ELISA at the same leave.