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A Report on the Death of Mixed Infection of HEV and HBV  [PDF]
Yonggang Qu, Yujian Zheng, Chuangfu Chen, Guangze Zhu, Ningyi Jin
Journal of Biosciences and Medicines (JBM) , 2015, DOI: 10.4236/jbm.2015.33015
Abstract:

Mixed infection with hepatitis E virus (HEV) in patients with chronic hepatitis B virus (HBV) infection is frequent. HEV mixed infection often leads to activation of hepatic pathological changes and worsens the inflammatory activity. However, it is not known clearly how these two types of virus influence each other in human body. Intensive investigation has revealed that HEV mixed infection inhibits HBV replication. We have just encountered a relative rare case. The patient who was a HBV carrier and was infected by HEV. Before he was infected by the HEV, the measurement of his HBV DNA fixed quantity examination on fluorescence was <103 copies/ml; his routine biochemistry was normal; and his anti HEV-IgM and anti-HEV-IgG appeared to be negative reaction. After he was infected by HEV, his routine biochemistry increased, and the measurement of his HBV DNA fixed quantity examination on fluorescence was 8.51 × 105 copies/ml. It indicated that the replication of HBV was activated after the patient infected HEV. Finally, he was dead. This case revealed that HEV mixed infection may activate the replication of HBV, not inhibit HBV replication, and demonstrated the needs for further studies about the mechanism of the interaction of the two viruses.

Molecular Characterization of msp2/p44 of Anaplasma phagocytophilum Isolated from Infected Patients and Haemaphysalis longicornis in Laizhou Bay, Shandong Province, China
Yong Wang, Chuangfu Chen, Lijuan Zhang
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0078189
Abstract: Molecular characterization of the MSP2/P44 protein of Anaplasma phagocytophilum may determine not only if the bacterium is capable of invading hosts but also whether it generates antigenic variation for the purpose of escaping the host immune response, resulting in various pathologic injuries and serious clinical outcomes. Chinese anaplasmosis patients usually present with serious manifestations, and the fatality rate is as high as 26.5%. In this study, we amplified, cloned and sequenced the msp2/p44 genes of three Chinese A. phagocytophilum isolates from Laizhou Bay, Shandong Province, where human granulocytic anaplasmosis (HGA) patients present severe clinical manifestations, and analyzed their genetic characterization and structural features. We also compared them with the HZ and Webster A. phagocytophilum strains. The sequences for both strains are available in GenBank. Analyses indicated that Chinese A. phagocytophilum isolates were significantly different from the HZ and Webster strains in terms of nucleotide sequences, amino acid sequences and protein secondary and tertiary structures. Moreover, the number of immunologic B-cell epitopes (19) of the MSP2 protein of the Chinese isolates was higher than that of the A. phagocytophilum strains HZ (16) and Webster (9). This genetic diversity of the MSP2/P44 protein of Chinese A. phagocytophilum isolates might be relevant and might have serious clinical outcomes. This observation could provide a clue to further understand the pathogenesis of Chinese A. phagocytophilum.
Inhibition of foot-and-mouth disease virus replication in vitro and in vivo by small interfering RNA
Wang Pengyan, Ren Yan, Guo Zhiru, Chen Chuangfu
Virology Journal , 2008, DOI: 10.1186/1743-422x-5-86
Abstract: Foot-and-mouth disease (FMD) is an acute and highly contagious disease requiring expensive treatment occurring in cloven-hoofed animals. The etiological agent of FMD is foot-and-mouth disease virus (FMDV), which belongs to the genus Aphthovirus of the family Picornaviridae [1]. The spreading capacity of the virus and its ability to change its antigenic identity make it a serious threat to the beef and dairy industries in many countries. FMDV has 7 serotypes and over 70 subtypes. Owing to the absence of reciprocal protection among all the serotypes, it is difficult to control FMD through vaccination and impossible to eliminate FMD by conservative natural breeding. A recent occurrence of a large epidemiogenesis has made the development of emergency antiviral strategies essential for preventing outbreaks of FMD.RNA interference (RNAi) is a process of sequence-specific, posttranscriptional gene silencing (PTGS) in animals and plants, which can be induced by 21- to 23-nucleotide (nt) siRNA that demonstrates sequence homology to the target gene [2,3]. It is well known that one obvious potential function for the RNAi machinery would be to defend cells against viruses that express dsRNA as part of their life cycle [4]. Indeed, there is compelling evidence indicating that RNAi is critical incurtailing viral infections in both plants and invertebrates. Moreover, it can be readily demonstrated that the artificial induction of an antiviral RNAi response in mammalian cells can confer strong protection against a wide range of pathogenic viruses [5]. Nevertheless, it remains unclear whether RNAi is involved in antiviral defense in mammalian cells in physiological conditions. Mammalian cells were originally thought to be unlikely to posses an active RNA-silencing machinery [6], besides a nonspecific, interferon mediated antiviral response mediated by dsRNA [7,8], especially by viral long (35-nt) dsRNA [9]. The recent description of RNAi in mammalian cells proved that the RNA silenc
Enhanced Muscle Growth by Plasmid-Mediated Delivery of Myostatin Propeptide
Shengwei Hu,Chuangfu Chen,Jingliang Sheng,Yufang Sun,Xudong Cao,Jun Qiao
Journal of Biomedicine and Biotechnology , 2010, DOI: 10.1155/2010/862591
Abstract: Myostatin is a member of the transforming growth factor beta (TGF-) superfamily that functions as a negative regulator of skeletal muscle development and growth. Myostatin blockade therefore offers a strategy for promoting muscle growth in livestock production without resorting to genetic manipulation. In this report, we examined the effect of myostatin inhibition by plasmid-mediated delivery of a mutant myostatin propeptide (MProD76A), a natural inhibitor of myostatin, on the growth performance of mice. A significant increase in skeletal muscle mass was observed after a single intramuscular injection of naked plasmid DNA encoding MProD76A into mice. Enhanced muscle growth occurred because of myofiber hypertrophy, but no cardiac muscle hypertrophy and organomegaly was observed in the mice after myostatin inhibition by plasmid-mediated MProD76A delivery. These results demonstrate a promising approach to enhancing muscle growth that warrants further investigation in domestic animals.
Sheep 7SK promoter for short hairpin RNA expression
Ni,Wei; Hu,Shengwei; Hazi,Wureli; Wang,Yuanzhi; Qiao,Jun; Yan,Ren; Chen,Chuangfu;
Electronic Journal of Biotechnology , 2012,
Abstract: background: gene silencing mediated by small interfering rna (sirna) has become a powerful biological tool for the regulation of gene expression. for the synthesis of sirna by vector-based expression systems, several mammalian small nuclear rna (snrna) promoters have been cloned and shown different transcriptional efficiencies. results: in this study, we identified a sheep 7sk snrna (s7sk) promoter based on the highly conserved polymerase iii promoter elements. promoter activity was measured by promoter-driven shrna expression to suppress expression of an exogenous reporter gene and endogenous sheep gene. conclusions: the knock down assay demonstrated that the s7sk induced more stronger inhibition effect than human u6 and h1 promoters. the use of this native sheep 7sk promoter for shrna expressionis an important component for development of rnai-based gene therapy and production of transgenic animals in sheep species.
Interactions of Target Proteins with the virB4 Gene from Bovine Embryo Trophoblast Cells Infected with Brucella abortus Screened Through a Yeast Two-Hybrid Assay
Zhang Hui,Wang Pengyan,Wang Yuanzhi,Guo Qian,Li Chengyao,Chen Chuangfu
Journal of Animal and Veterinary Advances , 2012, DOI: 10.3923/javaa.2012.1793.1799
Abstract: To screen for the interaction of target proteins with the virB4 gene in bovine embryo trophoblast cells that were infected with Brucella bacteria. A cell culture for bovine trophoblasts was established. The cDNA library of trophoblast cells infected with the vaccine strain RB51 was constructed and a Saccharomyces cerevisiae expression vector containing the gene virB4 was synthesized. Target proteins which interacted with virB4 were screened through a yeast two-hybrid system. The results showed that the cDNA library of bovine trophoblast cells infected with the vaccine RB51 strain was successfully constructed. A recombinant plasmid pGBKT7-virB4 was cloned successfully into the expression vector Y187. Thirteen different types of proteins that interacted with virB4 were screened through a yeast two-hybrid assay.
Comparison Between the Effects of Valproic Acid and Trichostatin A on in vitro Development of Sheep Somatic Cell Nuclear Transfer Embryos
Shengwei Hu,Wei Ni,Chuangfu Chen,Wujiafu Sai,Wureli Hazi,Zhirui He,Ren Meng,Jixing Guo
Journal of Animal and Veterinary Advances , 2012, DOI: 10.3923/javaa.2012.1868.1872
Abstract: The present study was carried out to assess and compare the effects of Valproic Acid (VPA) and Trichostatin A (TSA) on in vitro development of sheep Somatic Cell Nuclear Transfer (SCNT) embryos. The results showed that treatment of cloned sheep embryos with 4 mM VPA or 50 nM TSA for 24 h after activation could significantly improve blastocyst rate compared to the control (30.7 vs. 23.3 vs. 16.7%, respectively p<0.05). VPA treatment resulted in a significant higher blastocyst rate than that of TSA-treated group (p<0.05). Moreover, VPA treatment significantly increased (p<0.05) total cell number per blastocyst compared with the TSA treatment and control groups (78.8±9.3 vs. 69.6±9.7 vs. 64.1±8.6, respectively). Furthermore, VPA treatment increased expression of the development-related genes OCT4 and SOX2 in SCNT blastocysts. These results demonstrate that VPA may be more potent than TSA in supporting developmental competence of cloned embryos.
Interaction Between VirB5 of Brucella Type IV Secretion System (TFSS) and Ferritin Heavy Polypeptide 1 (FTH1) in Murine Macrophage
Fei Guo,Yuanzhi Wang,Chuangfu Chen,Hui Zhang,Jun Qiao,Yan Ren,Junbo Zhang,Zhiqiang Li
Journal of Animal and Veterinary Advances , 2012, DOI: 10.3923/javaa.2012.2623.2629
Abstract: TFSS is an important virulence factor of Brucella, organized as one operon containing 12 different across the cell wall bacterial proteins among which VirB5 regulates the host phagocytosis of Brucella and the transportation of Brucella in the host cells. This study has constructed cDNA library from Brucella melitensis 16M-infected murine macrophage Raw264.7, identified and confirmed the interaction between Brucella VirB5 and FTH1 of RAW264.7 using yeast two-hybrid and Co-Immunoprecipitation (Co-IP) technologies. Subsequently, the morphological changes and the expression of apoptosis-related genes in Brucella-infected RAW264.7 cells have been investigated with Electron Microscope (EM) and real-time quantitative RT-PCR, respectively. The present study has demonstrated that VirB5 and FTH1 play important roles in intracellular parasitism of Brucella and inhibition of FHT1 expression accelerates the apoptosis of macrophage.
Inhibition of Foot and Mouth Disease Virus Replication in vitro and in vivo by Dual Short Hairpin RNA-Mediated RNA Interference
Wureli Hazi,Shengwei Hu,Wei Ni,Zhirui He,Ren Meng,Chuangfu Chen,Ningying Xu
Journal of Animal and Veterinary Advances , 2012, DOI: 10.3923/javaa.2012.1527.1531
Abstract: Foot and Mouth Disease Virus (FMDV) is one of the most important pathogen to the cattle industry often resulting in severe economic losses. Researchers have reported previously a therapeutic application of plasmid-based shRNA against FMDV but the high degree of sequence diversity between different FMDV serotypes may result in the appearance of escape mutants. In this study, a dual shRNA expression plasmid which can simultaneously express two different shRNA molecules was established and showed stronger inhibitory effects on virus replication than the mixture of two shRNAs. Moreover, the antiviral activity induced by the dual shRNA expression system was aslo evident on other FMDV serotypes. Therefore, the dual shRNA system targeting two conserved regions of virus genome provides a more powerful strategy for inhibiting FMDV replication in a cross-resistance manner and implicates a potential application in the treatment of high genetic variability of FMDV.
Characterization of β-Actin Promoter of Leuciscus merzbacheri
Wenge Hu,Shiwei Ma,Chuangfu Chen,Yuanzhi Wang,Yan Ren,Xudong Cao,Jinliang Sheng
Journal of Animal and Veterinary Advances , 2012, DOI: 10.3923/javaa.2012.3849.3854
Abstract: Leuciscus merzbacheri is a unique vulnerable indigenous fish which is only distributed in the Junggar Basin, Xinjiang. In this study, researchers cloned two L. merzbacheri β-actin promoter fragments of different length: SZ11 and SZ21 and analyzed their structural features. The mammalian expression vector pEGFP-N1 was used to construct eukaryotic expression vectors β1 pEGFP-N1-AFP III and β2 pEGFP-N1-AFP III in which fish type III antifreeze protein gene was used as the structural gene. The results showed that the cloned two fragments of β-actin promoters had the ability to drive the expression of the green fluorescent protein gene in BHK-21 cells. These data suggest that the vectors researchers constructed based on β-actin promoter could be exploited as all fish recombinant eukaryotic expression vector.
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