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2019 Vol.14 No.4
Published 2019-08-25

Invited paper
Original Article
Case Analysis
Review
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Original Article
199 HU Kongying1, GU Chenjian1, WU Minle2, TAO Shuai1, LIU Nannan1, LIU Jing1, XIE Youhua1
Construction of Flag-GPI carrying recombinant hepatitis B virus vector
Hepatitis B virus (HBV) is a small hepatophilic DNA virus that can specifically infect human hepatic parenchymal cells. The susceptibility of HBV can be attributed to cell surface receptor(s), intracellular proteins and other factors. A simple and clear method to assay the susceptibility of cells for HBV remains a challenge. Here, we constructed the recombinant HBV vectors 5c3c-CD59-Flag-GPI and 5c3cT-Flag-GPI based on the original HBV 5c3c vector and glycosylphosphatidylinositol (GPI) anchor. After transfection into Huh7 cells, Flag tags can be anchored on the cell membrane, and sorted by flow cytometry using Flag antibody. With the replenishment of HBV envelope protein, 5c3cT-Flag-GPI can form complete recombinant HBV particles. This study laid a foundation for optimization of packaging efficiency of recombinant HBV, which would provide a tool for HBV susceptibility research.
2019 Vol. 14 (4): 199-208 [Abstract] ( 91 ) [HTML 1KB] [PDF 5953KB] ( 754 )
209 DAI Xiaoyi, ZHONG Jiang
Construction of miRNA-205-regulated recombinant Coxsackievirus B3 and its replication
A recombinant Coxsackievirus B3 (CVB3), v205T, was constructed by inserting repeats of a target sequence for miR-205 between the coding sequence of P1 and P2. Its replication was analyzed in Hela and A549 cells, in which the level of miRNA-205 was known to be low and high, respectively. The results showed that the insertion of miRNA-205 targets in the selected site did not affect the infectivity of the recombinant virus in Hela cells, but greatly reduced its replication in A549 cells, resulting in more than 100-fold reduction in virus titer. miR-205 mimics and inhibitors were used to study the mechanism of differential replication of v205T. miR-205 mimics inhibited the replication and cytotoxicity of v205T in Hela cells, whereas miRNA-205 inhibitors increased the replication and cytotoxicity of v205T in A549. These results showed that the replication of v205T was regulated by cellular miR-205. The work provided basis for using CVB3 vectors for cancer viral therapy and vaccine development.
2019 Vol. 14 (4): 209-215 [Abstract] ( 78 ) [HTML 1KB] [PDF 2719KB] ( 665 )
216 LI Xu, ZHAO Lin, FAN Fenxia, LI Zhe, LU Xin, PANG Bo, KAN Biao
Genomic structure and rearrangement of the lysogenic bacteriophage pre-CTXΦ in four non-toxigenic strains of Vibrio cholerae serogroups O1 and O139
The lysogenic phage CTXΦ of Vibrio cholerae can carry and transfer the cholera toxin gene ctxAB from toxigenic strains to non-toxigenic strains. CTXΦ also has a precursor, pre-CTXΦ,which does not carry ctxAB gene. In this study, we sequenced the genomes of four nontoxigenic serogroups O1 and O139 strains carrying pre-CTXΦ with the next generation sequencing. Both short-reading and long-reading sequencing were conducted to facilitate the assembly of long repeats. pre-CTXΦ prophage sequences were extracted from the genomes of these strains. The genes of CTXΦ receptor TcpA and the receptor-binding protein pIII in CTXΦ genome was subjected to sequence alignment, phylogenic tree construction and possible domains for protein-protein interactions. The tandem repeats of the different pre-CTXΦ alleles and their novel arrangement in the genomes were identified, and the precursor genome of classical type CTXΦ was also found. The arrangements of the pre-CTXΦ alleles in these strains were significantly different from the pandemic O1 El Tor strains. New sequences of the TcpA and the pIII were found in some strains. TcpA-pIII sequence correspondence analysis indicated the complicated interactions of the different sequence types of both proteins during the infection of pre-CTXΦ alleles. In addition, we have discovered a new type of pre-CTXΦ on small chromosome of VC702. The rstR-4 gene in VC702 has been disrupted by an insertion of a transposon which was firstly found in Klebsiella pneumoniae. Our study revealed new sequence types and arrangement of the pre-CTXΦ alleles and new evidence for better understanding the horizontal transfer of CTXΦ/pre-CTXΦ among the strains.
2019 Vol. 14 (4): 216-224 [Abstract] ( 78 ) [HTML 1KB] [PDF 3136KB] ( 708 )
 
Case Analysis
225 HU Qiankun, SONG Jie, JIANG Xuhua, WANG Qianqian, LI Qiang, HUANG Chenlu, XU Wei, CHEN Liang, HUANG Yuxian
A case of bloodstream Streptococcus gallolyticus infection in a decompensated cirrhotic patient
Streptococcus gallolyticus (S. gallolyticus) is a Gram-positive group D streptococcus, belonging to Streptococcus bovis/Streptococcus equinus complex (SBSEC). This bacterium is an opportunistic pathogen that can cause infections in multiple sites and organs. However, the reports of bloodstream infection caused by S gallolyticus in patients with decompensated cirrhosis are rare. In this case, the patient developed diarrhea and fever after eating unclean food. Blood culture was obtained during chills and fever, and a strain of Gram-positive bacteria was isolated. The biochemical methods and 16S rRNA gene sequencing were used to identify the bacterial strain as Streptococcus gallolyticus subsp. gallolyticus. According to the results of antimicrobial susceptibility test, the patient was given meropenem 0.5 g q8h for 7 days and levofloxacin 0.5 g qd for another 7 days. The patient’s temperature returned to normal after the treatment.
2019 Vol. 14 (4): 225-229 [Abstract] ( 77 ) [HTML 1KB] [PDF 593KB] ( 675 )
 
Review
230 ZHU Dong2, YANG Cheng2, TONG Shuping1, LIU Shaojun3, WANG Yongxiang1
Revisit the implications of kidney infection by hepatitis B virus
Hepatotropism of hepatitis B virus (HBV) infection depends on the specificity of interaction between the virus and its receptors, host factors required for covalently closed circular DNA (cccDNA) formation, and nuclear factors promoting viral RNA transcription. Such requirement for HBV infection most likely are met in human kidney. Furthermore, multiple studies have identified many, if not all, markers of HBV infection in kidney cells. This review discusses the possibility that HBV does establish infection in human kidney. Recently, loss of serum hepatitis B surface antigen (HBsAg) has become a key indicator of functional cure of chronic hepatitis B (CHB). Given that human kidney is another potential target organ supporting HBV infection, antigen expression, and genome replication, the roles of kidney in the functional cure of CHB should be taken into consideration.
2019 Vol. 14 (4): 230-237 [Abstract] ( 76 ) [HTML 1KB] [PDF 629KB] ( 591 )
238 YE Fei, ZHANG Xinxin, YU Demin
Epigenetic regulation of hepatitis B virus covalently closed circular DNA: current progress
Hepatitis B virus (HBV) infection remains a significant public health problem worldwide. Current therapies have made great progress for the controlling of liver disease progression and its detrimental clinical sequelae. However, the lack of curativeness of current antiviral therapies is attributed to their inability to eliminate HBV covalently closed circular DNA (cccDNA), which serves as a key factor in maintaining the persistent form of the viral genome and transcription template for virus reproduction, leading to chronic hepatitis B patients requiring long-term medication or relapse after treatment. Previous studies have confirmed that the transcription from HBV cccDNA is regulated by epigenetic mechanism, including cccDNA methylation, histone modification, microRNAs, chromatin remodeling and other aspects. In this review, we summarize the latest progress on epigenetic regulation of HBV from the related aspects.
2019 Vol. 14 (4): 238-245 [Abstract] ( 65 ) [HTML 1KB] [PDF 638KB] ( 672 )
246 LIU Lei, LI Qihan
Progress in herpes simplex virus type 2 vaccine research
Herpes simplex virus type 2 (HSV-2) is a member of the alpha subgroup of the human herpes virus family. It has a strong pathogenic ability to cause genital herpes and other herpes diseases in various age groups. Due to its complex genetic structure and pathological features, there is no effective clinical treatment for this medical concern. In addition, latent infection of nerve system, viral reactivation and viral induced immune suppression are major challengers for vaccine research and development.
2019 Vol. 14 (4): 246-251 [Abstract] ( 104 ) [HTML 1KB] [PDF 577KB] ( 810 )
252 ZHANG Ziling, LIU Mingbin, XU Jianqing, ZHANG Xiaoyan
Progress on broadly neutralizing antibodies against human immunodeficiency virus type 1
The morbidity and mortality caused by human immunodeficiency virus type 1 (HIV)/acquired immunodeficiency syndrome (AIDS) were significantly controlled through the combination of anti-HIV-1 therapy. However, HIV/AIDS still could not be cured because of the persistence of HIV-1 reservoir. The broadly neutralizing antibody (bNAb) can effectively reduce the HIV-1 viral load and slow down disease progress, which might be an effective strategy to control HIV/AIDS. The practice of sequential immune strategies has greatly promoted the development of bNAbs. In March 2018, US Food and Drug Administration (FDA) approved the first monoclonal bNAb for the treatment of adult with multidrug resistant HIV, which greatly inspired the enthusiasm for bNAb development. The present paper reviewed the research progress and challenging on bNAb development in recent years.
2019 Vol. 14 (4): 252-258 [Abstract] ( 89 ) [HTML 1KB] [PDF 596KB] ( 613 )
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