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2020 Vol.15 No.4
Published 2020-08-25

Invited paper
Original Article
Review
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Original Article
206 WANG Yingnan, YI Ming, XU Weizhen, ZHONG Zhaohua
Sequence analysis for the spliced fragments of Coxsackievirus B3 genome
The genomic RNA instability of Coxsackievirus B (CVB) occurs in the infected cells. The sequence characteristics and generating mechanism of these viral RNA fragments remain unknown. In this study, the genomic segments of CVB group B type 3 (CVB3) in infected cells were amplified, cloned with 5′ rapid amplification of cDNA ends (5′ RACE) assay and sequenced. The secondary structures of their 5′ ends were analyzed. The results showed that twenty fragments derived from CVB3 genome were obtained with lengths ranging from 2 067 to 5 547 bp. The 5′ ends of these fragments mainly located in the 2Apro- and 2C-coding regions. RNA folding analysis showed that the majority of these fragments could form secondary stem-loop structure in their 5′ ends. This study indicates that incomplete RNA fragments derived from CVB genome were abundant in the infected cells. These fragments could form a partial dsRNA in their 5′ ends. Our data suggests that these fragments are not generated randomly but more likely by the cleavage of RNA endonuclease. This observation is helpful to understand the mechanism for the genome instability of CVB.
2020 Vol. 15 (4): 206-212 [Abstract] ( 10 ) [HTML 1KB] [PDF 3172KB] ( 17 )
213 QIN Boyin, WANG Chao, LIU Yang, REN Xiaonan, FANG Zhong, LI Shun, ZHOU Xiaohui
Receptor-interacting protein kinase 3 plays a role in induction viral antigen specific CD8memory T cell responses against influenza A (H1N1) virus infection in mice
Receptor-interacting protein kinase 3 (RIPK3) is one of the key components of necrotic complex, which mediates the development of programmed cell necrosis. Our previous studies have revealed that the initial response of total influenza antigen-specific CD8T cells is partially dependent on RIPK3, but its role in memory CD8T cell response is unclear yet. In this study, we observed that the frequency of influenza specific CD8T cells and their ability to secrete cytokine IFN-γ and TNF-α in RIPK3 knockout mice were significantly lower than those in wild-type mice 37 days after the initial infection of H1N1 A/Puerto Rico/8/34 strain (PR8) in C57BL/6 mice. After the re-challenging infection of H1N1virus PR8, the frequency of viral specific CD8T cells and their ability to secrete cytokine IFN-γ in RIPK3 knockout mice were still significantly lower than those in wild-type mice. In addition, while the frequency of CD8central memory T cells (TCM) in RIPK3 knockout mice was significantly higher than that in wild-type mice, the proportion of effective memory T cells (TEM) or effective T cells (TEff) was significantly lower than that in wild-type mice. These results indicate that RIPK3 is involved in the regulation of the number and secretion of cytokines of influenza virus specific memory CD8T cells, and affects the ratio of TCM to TEM or TEff in memory CD8T cells, which provide new clues for further exploring the molecular mechanism of virus specific memory CD8T cell response. response.
2020 Vol. 15 (4): 213-219 [Abstract] ( 6 ) [HTML 1KB] [PDF 3018KB] ( 15 )
220 LIU Jia, WAN Simin, BAI Lu, LI Jianhua
Establishment of a method to effectively knock down gene expression in primary B cells to quickly identify functional gene
The process of B cell-mediated antiviral humoral immune response involves the up-regulation of genes. In order to identify the function of these genes quickly, we need a method to knock down target gene expression in B cells effectively in vitro and in vivo. In this study, four procedures were adopted to improve the assay. Firstly, we co-transfected the Drosha enzyme-specific siRNA with the retroviral packaging plasmid into virus packaging cells. Secondly, we constructed an in vitro culture system for primary B cell by adding anti-CD180 antibody to the culture medium, in which B cells can proliferate robustly. Then, by increasing the times of spin infection, the transduction efficiency of B cells was further improved. Besides, through pre-infection of mice, more proliferated and differentiated B cells after adoptive transfer can be harvested for phenotypic analysis. By adopting the above-mentioned improvement measures, the expression of B cell functional gene Bcl6 was successfully knocked down, and its anti-apoptotic function was verified. Establishment of this method would lay a good foundation for studying the mechanism of B cell proliferation, differentiation and defection in acute and chronic viral infections in the future.
2020 Vol. 15 (4): 220-227 [Abstract] ( 8 ) [HTML 1KB] [PDF 3709KB] ( 33 )
228 XUE Xiaojie, CHEN Meizhou, ZHANG Changliang, WANG Hongliang
The value of combined detection of new coronavirus IgM / IgG, nucleic acids and interleukin 6 in the diagnosis and treatment of new coronavirus disease
This article aims to explore the combined detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) immunoglobulin M (IgM)/immunoglobulin G (IgG), viral nucleic acid and interleukin 6 (IL-6) in the diagnosis and treatment of coronavirus disease in 2019 (COVID-19). This study collected 93 patients (51 critical, 18 severe, 15 mild and 9 normal) according to the standard of “New Coronavirus Pneumonia Diagnosis and Treatment Plan (Version 7)” and 20 suspected cases (negative nucleic acid test but clinical symptoms and Ct test results met the standards). 110 patients with other diseases such as pediatrics, gynecology, tumors, blood and digestion illness were selected as the control group. Automatic chemiluminescence immunoassay technology and electrochemiluminescence technology was used to detect 2019-nCoV IgM/IgG and IL-6 in the serum of all subjects. Real-time fluorescence quantitative reverse transcription polymerase chain reaction was used to detect SARS-CoV-2 nucleic acid in pharyngeal swabs of case group and control group. The results showed that the positive rates of serum IgM, IgG and IL-6 in suspected cases were 85%, 75% and 0%, respectively, and the positive rates in patients diagnosed with 2019 coronavirus were 98.9%, 95.7% and 75.2%, respectively (Among them, critical cases are 100%, 100% and 100%, severe cases are 94.4%, 100% and 97.9%, mild cases are 100%, 93.3% and 5%, and common types are 100%, 66.7% and 0%). The changes of IL-6 and 2019-nCoV IgM/IgG expression levels were related to the severity of the patients' disease, and the differences were statistically significant (χ2=273.51, χ2=149.37; P<0.05). The combined detection of serum IL-6 and 2019-nCoV IgM/IgG can be used as a monitoring indicator for the diagnosis and treatment of new coronavirus infection, and can also be used as an effective complement to the false negative detection of SARS-CoV-2 nucleic acid.
2020 Vol. 15 (4): 228-232 [Abstract] ( 8 ) [HTML 1KB] [PDF 505KB] ( 30 )
233 Xia Cai 1,Wendong Han 1,Zhiping Sun 1,Yun Qian 1,Li Chen 2,Youhua Xie 2,Di Qu1,2
Construction of emergency capability and connotation of BSL-3 laboratory from new coronavirus epidemic situation
Facing the outbreak of COVID-19, whether biosafety level 3 (BSL-3 ) laboratory can be immediately started in time and operate safely providing a reliable platform for COVID-19 virus research and detection, is ordeal for a laboratory's ability meeting with an emergency. After receiving the mission, BSL-3 laboratory of Fudan University immediately started the application process for new coronavirus experiment activities in the lab, following the regulations of China. After obtained the approval from the National Accreditation Service for Conformity Assessment and the National Health Commission, New coronavirus strains were isolated from a sample of Shanghai COVID-19 patient within one week, which reflecting the importance of laboratory’s connotation construction in peacetime. The BSL-3 laboratory’s connotation construction includes the biosafety management systems such as risk assessment and management of laboratory, personnel training, and management of facilities and equipment, ect. In this article, we shall introduce the experience and comprehension gained from management of the Fudan University BSL-3 laboratory.
2020 Vol. 15 (4): 233-240 [Abstract] ( 6 ) [HTML 1KB] [PDF 1060KB] ( 40 )
241 ZHAO Yamin, ZHU Shengling, WENG Shufeng, ZHANG Tianran, WANG Honghai, XU Ying
The effect of Mycobacterium tuberculosis protein Rv3425 in Mycobacterium smegmatis
Rv3425 encodes the PE/PPE family protein PPE57, which is a candidate antigen for vaccine construction that was tested in mice and rhesus monkeys in early studies of this laboratory. To study the function of Rv3425 of Mycobacterium tuberculosis (M. tuberculosis), Mycobacterium smegmatis (M. smegmatis) was selected as a model strain, and the recombinant strain Ms-Rv3425 was constructed. Compared with wild type, the colony morphology of Ms-Rv3425 was rougher and more bulging, and the ability of pellicle and aggregate formation was stronger. Ms-Rv3425 showed higher resistance to stress when cultured in acidic, sodium dodecyl sulfate (SDS), ampicillin, isoniazid, and rifampin conditions. The expression level of Rv3425 in the attenuated strain of M. tuberculosis H37Ra cultured under these adverse conditions was also significantly up-regulated. Infection of macrophages showed that Ms-Rv3425 enhanced the intracellular survival of M. smegmatis in THP-1 cell line. It led to higher mortality in BALB/c mice, and the bacterial retention and pathological damage of each organ were also higher than that of the control group. In summary, over expression of Rv3425 in M. smegmatis enhanced stress resistance, drug resistance and virulence.
2020 Vol. 15 (4): 241-250 [Abstract] ( 5 ) [HTML 1KB] [PDF 30648KB] ( 18 )
 
Review
251 ZHOU Wei,ZHANG Shaoyan,QIU Lei,HU Jun,ZHANG Huiyong,LU Zhenhui
New treatment advances in methicillin-resistant Staphylococcus aureus infection
Methicillin-resistant Staphylococcus aureus (MRSA) has become an increasingly invasive and prevalent pathogen. Due to chromosomal mediation, plasmid transfer, gene expression regulation, and active efflux systems, MRSA is resistant to a variety of antibiotics. When considering the issue from an ecological and evolutionary standpoint of view, it is evident that it is insufficient to solve the antibiotic dilemma only by antibiotics. This has forced people to look for a class of drugs that are completely different from existing antibiotics to treat MRSA. With the new information from immunology and biology, development of biological agents against MRSA infection have emerged. In this review, we summarize the latest advances in the treatment of MRSA infection with biologics, chemicals and traditional Chinese medicines.
2020 Vol. 15 (4): 251-258 [Abstract] ( 8 ) [HTML 1KB] [PDF 726KB] ( 15 )
259 LI Qian, MAO Xuhu
Progress in the mechanisms of Burkholderia pseudomallei infection and immune evasion
Burkholderia pseudomallei is the causative agent of melioidosis, which is classically characterized by pneumonia and multiple abscesses with a mortality rate up to 40%. There is growing evidence that it is considered a potential emerging infectious disease in many tropical developing countries. In this review, we highlight recently characterized mechanisms of remarkable virulence factors by which B. pseudomallei avoids killing by host immune responses.
2020 Vol. 15 (4): 259-264 [Abstract] ( 10 ) [HTML 1KB] [PDF 2583KB] ( 17 )
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