Antibodies elicited by Newcastle disease virus-vectored H7N9 avian influenza vaccine are functional in activating the complement system

H7N9 subtype avian influenza virus poses a great challenge for poultry industry.Newcastle disease virus(NDV)-vectored H7N9 avian influenza vaccines(NDV_(vec)H7N9)are effective in disease control because they are protective and allow mass administration.Of note,these vaccines elicit undetectable H7N9-specific hemagglutination-inhibition(HI)but high IgG antibodies in chickens.However,the molecular basis and protective mechanism underlying this particular antibody immunity remain unclear.Herein,immunization with an NDV_(vec)H7N9 induced low anti-H7N9 HI and virus neutralization titers but high levels of hemagglutinin(HA)-binding IgG antibodies in chickens.Three residues(S150,G151 and S152)in HA of H7N9 virus were identified as the dominant epitopes recognized by the NDV_(vec)H7N9 immune serum.Passively transferred NDV_(vec)H7N9 immune serum conferred complete protection against H7N9 virus infection in chickens.The NDV_(vec)H7N9 immune serum can mediate a potent lysis of HA-expressing and H7N9 virus-infected cells and significantly suppress H7N9 virus infectivity.These activities of the serum were significantly impaired after heat-inactivation or treatment with complement inhibitor,suggesting the engagement of the complement system.Moreover,mutations in the 150-SGS-152 sites in HA resulted in significant reductions in cell lysis and virus neutralization mediated by the NDV_(vec)H7N9 immune serum,indicating the requirement of antibody-antigen binding for complement activity.Therefore,antibodies induced by the NDV_(vec)H7N9 can activate antibody-dependent complement-mediated lysis of H7N9 virus-infected cells and complement-mediated neutralization of H7N9 virus.Our findings unveiled a novel role of the complement in protection conferred by the NDV_(vec)H7N9,highlighting a potential benefit of engaging the complement system in H7N9 vaccine design.

HBeAg gene expression with baculovirus vector in silk worm cells

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Characteristics and advantages of adeno-associated virus vector-mediated gene therapy for neurodegenerative diseases

Common neurodegenerative diseases of the central nervous system are characterized by progressive damage to the function of neurons, even leading to the permanent loss of function. Gene therapy via gene replacement or gene correction provides the potential for transformative therapies to delay or possibly stop further progression of the neurodegenerative disease in affected patients. Adeno-associated virus has been the vector of choice in recent clinical trials of therapies for neurodegenerative diseases due to its safety and efficiency in mediating gene transfer to the central nervous system. This review aims to discuss and summarize the progress and clinical applications of adeno-associated virus in neurodegenerative disease in central nervous system. Results from some clinical trials and successful cases of central neurodegenerative diseases deserve further study and exploration.

Construction and characterization of calreticulin-HBsAg fusion gene recombinant adenovirus expression vector

AIM: To generate recombinant adenoviral vector con-taining calreticulin (CRT)-hepatitis B surface antigen (HBsAg) fusion gene for developing a safe, effective and HBsAg-specific therapeutic vaccine.METHODS: CRT and HBsAg gene were fused using polymerase chain reaction (PCR), endonuclease diges-tion and ligation methods. The fusion gene was cloned into pENTR/D-TOPO transfer vector after the base pairs of DNA (CACC) sequence was added to the 5′ end. Adenoviral expression vector containing CRT-HBsAg fusion gene was constructed by homologous recombinan-tion. The human embryo kidney (HEK) 293A cells were transfected with linearized DNA plasmid of the recombi-nant adenoviral vector to package and amplify recombi-nant adenovirus. The recombinant adenovirus titer was characterized using the end-dilution assay. The expres-sion of the CRT/HBsAg fusion protein in Ad-CRT/HBsAg infected 293A cells was detected by Western blotting.RESULTS: The CRT-HBsAg fusion gene was char-acterized by PCR and sequencing and its length and sequence were confirmed to be accurate. The CRT-HB-sAg fusion gene recombinant pENTR/D-TOPO transfer vector was constructed. The recombinant adenoviral vector, Ad-CRT/HBsAg, was generated successfully. The titer of Ad-CRT/HBsAg was characterized as 3.9 × 1011 pfu/mL. The CRT-HBsAg fusion protein was ex-pressed by HEK 293A cells correctly. CONCLUSION: CRT/HBsAg fusion gene recombinant replication-defective adenovirus expression vector is constructed successfully and this study has provided an experimental basis for further studies of Hepatitis B vi-rus gene therapy.

Recombinant Vaccinia Virus is an Effective and Non-perturbing Vector for Human Dendritic Cells Transfected with Epstein-Barr Virus Latent Membrane Protein 2A

ObjectiveTo study the effects of dendritic cells (DC) transfected with recombinant vaccinia virus encoding Epstein Barr virus (EBV) latent membrane protein 2A(LMP2A) gene,and to provide evidence for further investigation on the therapeutic vaccines against EBV associated malignancies. MethodsMature DC were transfected with EBV LMP2A recombinant vaccinia virus (rVV LMP2A). Before and after the transfection,the expression of surface antigens on mature DC including CD1a,CD83,CD40,CD80,HLA DR was measured by fluorescence activated cell sorter (FACS) and the function of DC to stimulate allogeneic T cells proliferation was measured by mixed leukocyte reactions (MLR). ResultsLMP2A protein was highly expressed (66.1 %) in DC after the transfection of rVV LMP2A. No significant changes in the primary surface antigens expression and in the MLR were detected during the transfection. Transfected DC still had strong potential in stimulating the proliferation of allogeneic T cells. ConclusionRecombinant vaccinia virus was an effective and non perturbing vector to mediate the transfection of LMP2A into DC. The functions of mature DC were not affected significantly by the transfection of Vac LMP2A. This study could provide evidence for the further immunotherapy of EBV associated malignancies,e.g. nasopharyngeal carcinoma (NPC).

Evaluation of the Protection against Infectious Bursal Disease (IBD) Challenge in Progeny Born to Parents Having Received a Vaccination Program Using a Herpesvirus of Turkey-Infectious Bursal Disease (HVT-IBD) Vector Vaccine

Broiler breeder vaccination against IBD is usually based on the injection of at least one inactivated vaccine in oil adjuvant, typically included in a combined vaccine. Priming using one or several IBD vaccine (s) has been the most common way to immunize the breeders so far. In summary, protection against vvIBD challenge in chicks of one commercial genetic line vaccinated in ovo with the HVT-IBD vector vaccine was demonstrated. The parents’ IBD vaccination program, using the HVT-IBD vector vaccine alone, the HVT-IBD vector vaccine plus IBD inactivated vaccine, and inactivated IBD vaccine alone, did not impair their progeny’s in ovo HVT-IBD vector vaccine take and subsequent protection against vvIBD virus challenge. An advantage in terms of immunization of the progeny against vvIBD was shown in the chicks born to breeders vaccinated with the HVT-IBD vaccine as a primer, as compared to breeders vaccinated with the inactivated vaccine alone. High level of IBD maternally-derived antibodies transmitted to the progeny by their parents induces together with an early onset of immunity by in ovo injection of a HVT-IBD vector vaccine clinical protection, as monitored on bursas, after vvIBD virus challenge.

In situ tumor vaccination with adenovirus vectors encoding measles virus fusogenic membrane proteins and cytokines

AIM: To evaluate whether intratumoral expression of measles virus fusogenic membrane glycoproteins H and F (MV-FMG), encoded by an adenovirus vector Ad.MV-H/ F, alone or in combination with local coexpression of cytokines (IL-2, IL-12, IL-18, IL-21 or GM-CSF), can serve as a platform for inducing tumor-specifi c immune responses in colon cancer. METHODS: We used confocal laser scanning microscopy and flow cytometry to analyze cell-cell fusion after expression of MV-FMG by dye colocalization. In a syngeneic bilateral subcutaneous MC38 and Colon26 colon cancer model in C57BL/6 and BALB/c mice, we assessed the effect on both the directly vector-treated tumor as well as the contralateral, not directly vector- treated tumor. We assessed the induction of a tumor- specific cytotoxic T lymphocyte (CTL) response with a lactate dehydrogenase (LDH) release assay. RESULTS: We demonstrated in vitro that transduction of MC38 and Colon26 cells with Ad.MV-H/F resulted in dye colocalization, indicative of cell-cell fusion. In addition, in the syngeneic bilateral tumor model we demonstrated a signifi cant regression of the directly vector-inoculated tumor upon intratumoral expression of MV-FMG alone or in combination with the tested cytokines. We observed the highest anti-neoplastic efficacy with MV-FMG and IL-21 coexpression. The degree of tumor regression of the not directly vector-treated tumor correlated with the anti-neoplastic response of the directly vector-treated tumor. This regression was mediated by a tumor-specifi c CTL response. CONCLUSION: Our data indicate that intratumoral expression of measles virus fusogenic membraneglycoproteins is a promising tool both for direct tumor treatment as well as for tumor vaccination approaches that can be further enhanced by cytokine coexpression.

Comparison of Five Expression Vectors for the Ha Gene in Constructing a DNA Vaccine for H6N2 Influenza Virus in Chickens

A number of eukaryotic expression vectors have been developed for use as DNA vaccines. They showed varying abilities to initiate immune responses;however, there is little data to indicate which of these vectors will be the most useful and practical for DNA vaccines in different species. This report examines the use of five expression vectors with different promoters and Kozak sequence to express the same hemagglutinin (HA) protein of an H6N2 avian influenza virus for DNA vaccination in chickens. Although intramuscular vaccination with seven DNA constructs elicited no or limited measurable H6 HA antibody responses in Hy-Line chickens, variable reduction in virus shedding for either oropharyngeal or cloacal swabs post-virus challenge were observed. This indicated that all DNA constructs generated some levels of protective immunity against homologous virus challenge. Interestingly, lower dose (50 or 100 μg) of plasmid DNAs consistently induced better immune response than higher dose (300 or 500 μg). In the transfection experiments there appeared to be a hierarchy in the in vitro expression efficiency in the order of pCAG-optiHAk/ pCAG-HAk > pCI-HAk > VR-HA > pCI-HA > pCI-neo-HA > pVAX-HA. Since the level of in vitro expression correlates with the level of immune response in vivo, in vitro expression levels of the DNA constructs can be used as an indicator for pre-selection of plasmid vaccines prior to in vivo assessment. Moreover, our results suggested that the Kozak sequence could be used as an effective tool for DNA vaccine design.

Ecological Fitness of Non-vector Planthopper Sogatella furcifera on Rice Plants Infected with Rice Black Streaked Dwarf Virus

We evaluated the effects of rice black streak dwarf virus （RBSDV）-infested rice plants on the ecological parameters and its relevant defensive and detoxification enzymes of white-backed planthopper （WBPH） in laboratory for exploring the relationship between RBSDV and the non-vector planthopper. The results showed that nymph survival rate, female adult weight and fecundity, and egg hatchability of WBPH fed on RBSDV-infested rice plants did not markedly differ from those on healthy plants, whereas the female adult longevity and egg duration significantly shortened on diseased plants. Furthermore, significantly higher activities of defensive enzymes （dismutase, catalase and peroxidase） and detoxification enzymes （acetylcholinesterase, carboxylesterase and glutathione S-transferase） were found in WBPH adults fed on infected plants. Results implied that infestation by RBSDV increased the ecological fitness of non-vector planthopper population.

Construction of a replication-competent hepatitis B virus vector carrying secreted luciferase transgene and establishment of new hepatitis B virus replication and expression cell lines

BACKGROUND Previously,we have successfully constructed replication-competent hepatitis B virus(HBV)vectors by uncoupling the P open reading frame(ORF)from the preC/C ORF to carefully design the transgene insertion site to overcome the compact organization of the HBV genome and maintain HBV replication competence.Consequently,the replication-competent HBV vectors carrying foreign genes,including pCH-BsdR,carrying blasticidin resistance gene(399 bp),and pCH-hrGFP,carrying humanized renilla green fluorescent protein gene(720 bp),were successfully obtained.However,the replication efficiency of the former is higher but it is tedious to use,while that of the latter is poor and cannot be quantified.Hence,we need to search for a new reporter gene that is convenient and quantifiable for further research.AIM To establish a helpful tool for intracellular HBV replication and anti-viral drugs screening studies.METHODS We utilized the replication-competent HBV viral vectors constructed by our laboratory,combined with the secreted luciferase reporter gene,to construct replication-competent HBV vectors expressing the reporter gene secretory Nanoluc Luciferase(SecNluc).HepG2.TA2-7 cells were transfected with this vector to obtain cell lines with stably secreted HBV particles carrying secNluc reporter gene.RESULTS The replication-competent HBV vector carrying the SecNluc reporter gene pCHsNLuc could produce all major viral RNAs and a full set of envelope proteins and achieve high-level secreted luciferase expression.HBV replication intermediates could be produced from this vector.Via transfection with pTRE-sNLuc and selection by hygromycin,we obtained isolated cell clones,named HBV-NLuc-35 cells,which could secrete secNLuc recombinant viruses,and were sensitive to existing anti-HBV drugs.Using differentiated HepaRG cells,it was verified that recombinant HBV possessed infectivity.CONCLUSION Our research demonstrated that a replication-competent HBV vector carrying a secreted luciferase transgene possesses replication and expression ability,and the established HBV replication and expression cell lines could stably secrete viral particles carrying secNluc reporter gene.More importantly,the cell line and the secreted recombinant viral particles could be used to trace HBV replication or infection.

Non-transmissible Sendai virus vector encoding c-myc suppressor FBP-interacting repressor for cancer therapy

AIM:To investigate a novel therapeutic strategy to target and suppress c-myc in human cancers using far up stream element(FUSE)-binding protein-interacting repressor(FIR).METHODS:Endogenous c-Myc suppression and apoptosis induction by a transient FIR-expressing vector was examined in vivo via a HA-tagged FIR(HA-FIR)expression vector.A fusion gene-deficient,non-transmissible,Sendai virus(SeV)vector encoding FIR cDNA,SeV/dF/FIR,was prepared.SeV/dF/FIR was examined for its gene transduction efficiency,viral dose dependency of antitumor effect and apoptosis induction in HeLa(cervical squamous cell carcinoma)cells and SW480(colon adenocarcinoma)cells.Antitumor efficacy in a mouse xenograft model was also examined.The molecular mechanism of the anti-tumor effect and c-Myc suppression by SeV/dF/FIR was examined using Spliceostatin A(SSA),a SAP155 inhibitor,or SAP155siRNA which induce c-Myc by increasing FIR△exon2 in HeLa cells.RESULTS:FIR was found to repress c-myc transcription and in turn the overexpression of FIR drove apoptosis through c-myc suppression.Thus,FIR expressing vectors are potentially applicable for cancer therapy.FIR is alternatively spliced by SAP155 in cancer cells lacking the transcriptional repression domain within exon 2(FIR△exon2),counteracting FIR for c-Myc protein expression.Furthermore,FIR forms a complex with SAP155 and inhibits mutual well-established functions.Thus,both the valuable effects and side effects of exogenous FIR stimuli should be tested for future clinical application.SeV/dF/FIR,a cytoplasmic RNA virus,was successfully prepared and showed highly efficient gene transduction in in vivo experiments.Furthermore,in nude mouse tumor xenograft models,SeV/dF/FIR displayed high antitumor efficiency against human cancer cells.SeV/dF/FIR suppressed SSA-activated c-Myc.SAP155 siRNA,potentially produces FIR△exon2,and led to c-Myc overexpression with phosphorylation at Ser62.HA-FIR suppressed endogenous c-Myc expression and induced apoptosis in HeLa and SW480 cells.A c-myc transcriptional suppressor FIR expressing SeV/dF/FIR showed high gene transduction efficiency with significant antitumor effects and apoptosis induction in HeLa and SW480 cells.CONCLUSION:SeV/dF/FIR showed strong tumor growth suppression with no significant side effects in an animal xenograft model,thus SeV/dF/FIR is potentially applicable for future clinical cancer treatment.

Hepatitis B virus envelope as a targeting gene transfer vector for hepatic cancer cells

学习的目的是观察肝炎 B 病毒信封(HBVE ) 的 transfection 功效并且为肝癌症房间作为基因转移向量评估它的能力的目的。获得 HBVE 的方法， HepG 2.2.15 房间的上层清液液体与集中的一个 PEG8000 解决方案被混合并且被 β-propiolactone 使失去活性。获得的 HBVE 被用来包装 pIRES 2-EGFP 测试它的包裹能力。然后，我们与 ELISA， PCR， SDS 页和电子显微镜学检验了它的数量和质量。pIRES 2-EGFP 与 HBVE 被包装并且获得了产品 HBVE-GFP。pIRES 2-EGFP 被包装与脂肪一些并且获得了产品 liposome-GFP。HBVE-GFP 和 liposome-GFP 被用来转移 HepG 2 房间学习 transfection 效率。HBVE-GFP 被用来转移学习指向的能力的 HepG 2， A549， HeLa 和 FB 房间。绿荧光灯的蛋白质(GFP ) 表示在一台荧光灯的显微镜下面被观察。GFP 积极房间的率被流动 cytometry 决定。结果 1。获得的 HBVE 能保留表面蛋白质 HBsAg + pre S1 + pre S2 并且没有病毒 DNA。它为 pIRES 2-EGFP 有好包裹能力。2。Transfection 效率：GFP 能在荧光灯的显微镜下面在脂肪的一些组和 HBVE 组被观察。但是 HBVE 组比脂肪的一些组有更高荧光灯的紧张。脂肪的一些组的 transfection 率是 49.97% ± 2.37% 当 HBVE 组是 70.65% ±时 3.15% 并且 HBVE 组的荧光灯的紧张是 3 4 次(P = 0.000 ) 为有流动 cytometry 的决心的脂肪的一些组。3。指向能力：GFP 能在荧光灯的显微镜下面在四个组被观察。HepG 2 组在四个组之中有最高荧光灯的紧张。HepG 2 组的 transfection 率是 71.35% ± 0.03% 它高度比另外的组被表示(P = 0.000 ) 并且 HepG 2 组的荧光灯的紧张是 2 3 次(P = 0.000 ) 为有流动 cytometry 的决心的另外的 3 个组。结论 HBVE 能从 HepG 2.2.15 房间的上层清液液体与 PEG8000 和 β-propiolactone 的方法成功地被构造。HBVE 能是为肝癌症房间的候选人基因转移向量。

Complex interactions among insect viruses-insect vector-arboviruses

Insects are the host or vector of diverse viruses including those that infect vertebrates,plants,and fungi.Insect viruses reside inside their insect hosts and are vertically transmitted from parent to offspring.The insect virus-host relationship is intricate,as these viruses can impact various aspects of insect biology,such as development,reproduction,sex ratios,and immunity.Arthropod-borne viruses(arboviruses)that cause substantial global health or agricultural problems can also be vertically transmitted to insect vector progeny.Multiple infections with insect viruses and arboviruses are common in nature.Such coinfections involve complex interactions,including synergism,dependence,and antagonism.Recent studies have shed light on the influence of insect viruses on the competence of insect vectors for arboviruses.In this review,we focus on the biological effects of insect viruses on the transmission of arboviruses by insects.We also discuss the potential mechanisms by which insect viruses affect the ability of hosts to transmit arboviruses,as well as potential strategies for disease control through manipulation of insect viruses.Analyses of the interactions among insect vectors,insect viruses and arboviruses will provide new opportunities for development of innovative strategies to control arbovirus transmission.

Cloning and expression of core gene cDNA of Chinese hepatitis C virus in cosmid pTM3

AIM To clone core gene cDNA of Chinesehepatitis C virus(HCV)into eukaryoticexpression vector cosmid pTM3 and to expressHCV core antigen in HepG2 cells.METHODS Core gene cDNA of HCV wasintroduced into eukaryotic expression vectorcosmid pTM3.Using vaccinia virus/bacteriophage T7 hybrid expression system,HepG2 cells were transfected with therecombinant plasmid pTM3-Q534 by lipofectin.RESULTS From the transfected bacteriaTop10F’,2 pTM3-Q534 clones containing therecombinant plasmid were identified fromrandomly selected 10 ampicillin-resistantcolonies.By reverse transcription PCR andindirect immunofluorescence technique,HCVRNA and core protein was identified in HepG2cells transfected with the recombinant plasmid.CONCLUSION The construction of arecombinant plasmid and the expression of coregene cDNA of HCV in HepG2 was successful.

Aphids and their transmitted potato viruses: A continuous challenges in potato crops

Aphid is one of the most destructive insect pests on cultivated plants in temperate regions.Their piercing-sucking mouthparts and phloem feeding behavior directly damage crops and deplete plant nutrients.Potato(Solanum tuberosum L.)is one of the most important food sources on the planet,and several aphid species,e.g.,Myzus persicae(Sulzer)(green peach aphid)and Macrosiphum euphorbiae(Thomas)(potato aphid)(Hemiptera:Aphididae)colonize potato and transmit several economically important viruses.Aphid-transmitted potato viruses have been emerging all over the world as a very serious problem in potato production,inducing a wide variety of foliar and tuber symptoms,leading to severe yield reduction and loss of tuber quality.In this review,recent advances in understanding the interactions of potato viruses with their hosts,aphid vectors and the environment are described.

Seasonal abundance and potential of Japanese encephalitis virus infection in mosquitoes at the nesting colony of ardeid birds,Thailand

Objective:To investigate the abundance and seasonal dynamics of mosquitoes,and to detect Japanese encephalitis virus(JEV)in these mosquitoes at the nesting colony of ardeid birds.Methods:Mosquitoes were collected bimonthly from July 2009 to May 2010 by Centers for Disease Control.Light traps and dry ice,as a source of CO_2,were employed to attract mosquitoes.Mosquitoes were first identified,pooled into groups of upto 50 mosquitoes by species,and tested for JEV infection by viral isolation and reverse transcriptase polymerase chain reaction.Results:A total of 20370 mosquitoes comprising 14 species in five genera were collected.The five most abundant mosquito species collected were Culex tritaeniorhynchus(95.46%),Culex vishnui(2.68%),Culex gelidus(0.72%),Anopheles peditaeniatus(0.58%)and Culex quinquefasciatus(0.22%).Mosquito peak densities were observed in July.All of 416 mosquito pools were negative for JEV.Conclusions:This study provides new information about mosquito species and status of JEV infection in mosquitoes in Thailand.Further study should be done to continue a close survey for the presence of this virus in the ardeid birds.

Targeting adeno-associated virus and adenoviral genetherapy for hepatocellular carcinoma

Human hepatocellular carcinoma(HCC)heavily endangers human heath worldwide.HCC is one of most frequent cancers in China because patients with liver disease,such as chronic hepatitis,have the highest cancer susceptibility.Traditional therapeutic approaches have limited efficacy in advanced liver cancer,and novel strategies are urgently needed to improve the limited treatment options for HCC.This review summarizes the basic knowledge,current advances,and future challenges and prospects of adeno-associated virus(AAV)and adenoviruses as vectors for gene therapy of HCC.This paper also reviews the clinical trials of gene therapy using adenovirus vectors,immunotherapy,toxicity and immunological barriers for AAV and adenoviruses,and proposes several alternative strategies to overcome the therapeutic barriers to using AAV and adenoviruses as vectors.

Zika virus infection, transmission, associated neurological disorders and birth abnormalities: A review of progress in research, priorities and knowledge gaps

On February 1, 2016, the World Health Organization declared that the cluster of microcephaly cases and other neurological disorders constitute public health emergency of international concern. Furthermore, few studies demonstrated that there was an increased evidence of causal relationship of Zika virus(ZIKAV) infection and microcephaly, birth abnormalities and neurological disorders such as Guillain–Barre′ syndrome.ZIKAV transmission occurs mainly by the bite of infected mosquitos(Aedes species), but there are also reports that infections could occur via the placenta, breast milk, saliva,blood transfusion and sex. This article reviews the global efforts, progress in scientific research to understand the pathogenesis of ZIKAV infection & disease, clinical presentations, congenital transmission and autoimmune neurological disorders. The paper further explores the knowledge gaps, future priority research agenda for strategic response including vector control and prevention. We conducted a systematic literature review to synthesise available evidence on ZIKAV infection and its vector and host interaction from electronic databases.

Construction and Characterization of a Hepatitis B Virus Replicon

建立复制细胞肝炎 B 病毒(HBV ) 当模特儿并且在抗病毒的药评估决定它的应用程序，我们构造了表情包含了 HBV 染色体的 1.3 个拷贝，并且在 Huh7 房间在短暂 transfection 以后测量了病毒的复制的水平的 plasmid。我们然后观察了抗病毒的药管理的效果。HBV (ayw ) 基因碎片的 1.3 褶层被 PCR 和限制 endonuclease 消化克隆进 pCR2.1。recombinant plasmid 是进 Huh7 房间， HBsAg， HBeAg 和 HBV 的短暂 transfected 在 Huh7 房间的上层清液的 DNA 被 ELISA 和即时 PCR 分别地测量；细胞内部的 HBV replicative 中介和细胞内部的 HBV 抄本被南部的污点和北污点分别地检测。adefovir 的抗病毒的效果，新奇 anti-HBV 核苷酸类似物，在这个细胞的模型系统被评估。结果显示 HBV replicon 的 recombinant plasmid 成功地被构造；在 plasmid pHBV1.3 带的 HBV 染色体能高效地复制并且被表示在哈 7 个房间， adefovir 能在这个细胞的模型，和抑制禁止 HBV 复制是剂量依赖者。结论是 HBV replicon，能在 hepatoma 房间高效地开始病毒的复制，可以是在 HBV 复制和抗病毒的药的学习的一个有用工具。关键词肝炎 B 病毒 - 传染 replicon - 表示向量 CLC 数字 R373 基础条款：国家自然科学基础(No.30271170， No.30170889 ) 。

Adenovirus-Mediated Herpes Simplex Virus Thymidine Kinase Gene Transfer Driver by KDR Promoter in Treatment of Experimental Human HepatocelLular Carcinoma in Nude Mice

Objective： To investigate the therapeutic efficacy of adenovirus-mediated herpes simplex virus thymidine kinase （HSV-tk） gene transfer under the driving of KDR promoter （AdKDR-tk） in combination of ganciclovir （GCV） against human hepatocellular carcinoma in nude mice. Methods： HepG2 cell line was implanted subcutaneously into 32 nude mice, which were subsequently divided into 4 groups （n=8 each group）： Ganciclovir group （Ⅰ）, Ad group （Ⅱ）, AdCMV-tk/GCV group （under the driving of CMV promoter） （Ⅲ） and AdKDR-tk/GCV group （Ⅳ）. Then intratumoral injection of recombinant adenovirus or Ad was performed in all nude mice, and repeated 24 h later. For the following 10 d GCV was given at a dose of 100 mg/（kg· d）, ip. All the treated animals were killed to evaluate the tumor weight and the histopathological changes and the microvessel density of tumors after the treatment was determined. Results Compared with group Ⅰ, the tumor inhibitory rate was 12.3% in group Ⅲ and 24.5% in group Ⅳ; the inhibition rates were significantly different between group Ⅲand Ⅳ （P〈0.05）. The mean MVDs in group Ⅰ, Ⅱ, Ⅲ and Ⅳ were 37.4±8.6, 30.6±7.8, 27.6±7.1, and 10.7±4.1 （microvessels/mm^2）, respectively. Significant differences were found between group Ⅲ and Ⅱ （P〈0.05）, Ⅳ and Ⅱ （P〈0.01）, and Ⅳ and Ⅲ （P〈0.01）. Conclusion： Intratumoral injection of AdKDR-tk results in marked inhibition of HCC growth through inhibition angiogenesis in nude mice. It may be a new treatment approach for human HCC,