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Development of Cell Lines Stably Expressing Staphylococcal Nuclease Fused to Dengue 2 Virus Capsid Protein for CTVI

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Acta Biochimica et Biophysica Sinica 2004年第8期36卷 577-582页

作者： Cheng-Feng QIN and E-De QIN institute of microbiology and epidemiology, Academy of Military medical Sciences, Beijing 100071, China institute of microbiology and epidemiology Academy of Military Medical Sciences

To explore the potential application of capsid-targeted viral inactivation (CTVI) strategy in prophylactic model against dengue virus (DV) infection, here we fused a Ca2+-dependent nuclease, staphylococcal nuclease (SN), to the capsid protein of dengue 2 virus (D2C) at the carboxyl terminal, and constructed the desired expression plasmid pc/D2C-SN and control plasmids pc/D2C-SN* and pc/D2C. A mammalian cell line BHK-21 was transfected by electroporation with those plasmids and thereafter selected by 5 μg/ml blasticidin. The resistant cell clones were then expanding cultured and screened by RT-PCR and Western Blot assays. The nuclease activity of the expressed fusion protein D2C-SN was analyzed by in vitro DNA digestion assay. It was confirmed cell lines stably expressing D2C-SN and control constructs were obtained. The intracellular expressed fusion protein D2C-SN had ideal nuclease activity and no cytotoxicity on mammalian cells. Those engineered cell lines provided the experimental system for CTVI application in prophylactic model and paved the new road for combating DV infection with CTVI.

关键词： dengue 2 virus capsid protein staphylococcal nuclease CTVI stable expression

Study on the determinants of suckling mice neurovirulence of dengue 2 virus

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Science China(Life Sciences) 2003年第1期46卷 95-103页

作者：赵卫范宝昌胡志君陈水平王鹏程苑锡同李晓萸于曼秦鄂德杨佩英 institute of microbiology & epidemiology Academy of Military Medical Sciences

pDVWS501 was a genomic-length cDNA clone of dengue 2 virus, through which infec-tious virus (MON501) could be rescued. MON501 was neurovirulent in mice, whose E residues 62 and 203 were Lys and Asn, respectively. Two genomic-length cDNA clones (TB62 and TB203) were constructed by pointed mutation of pDVWS501 with OL-PCR, E62 of TB62 and E203 of TB203 were converted to Glu and Asp, respectively. RNA transcripts of pDVWS501, TB62 and TB203 were produced in vitro and electroporated into BHK-21 cells. The cultures were collected after 7 days and used as inoculum to infect C6/36 cells. The existence of rescued dengue viruses in the culture was proved by RT-PCR, and the typical cytopathic effect (CPE) of C6/36 caused by dengue virus emerged after 2—5 days?inoculation. Sequence analysis further confirmed the exis-tence of recovered and recombinant DEN2 viruses, whose 5′ termini had an additional non-virus nucleotides 揋? while the 3′ terminal sequences remained the same as natural. The neuroviru-lence of three viruses was evaluated in 1-day-old mice by the intracerebral route with 105—102 TCID50. Compared with MON501 group, the number of infected mice with the signs of encephalitis in HFT62 and HFT203 groups was less, and the surviving time was longer. The properties of these mutants demonstrated that E62 and E203 are determinants of suckling mice neurovirulence.

关键词： dengue 2 virus, virulent determinants, infectious transcripts, point mutation.

Pandora of Ebola virus: are we ready?

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Chinese Science Bulletin 2014年第32期59卷 4235-4236页

作者： Ruifu Yang Beijing institute of microbiology and epidemiology

在 2014 年 3 月 23 日，畿尼的健康的部通知了在东南的 forested 区域的 Ebola 病毒疾病(EVD ) 的很快传播的爆发，并且 4 ? 天以后，疾病被证实被传给 Conakry，畿尼的首都。疾病很快也传播了到利比里亚，尼日利亚和塞拉利昂。2014 年... 详细信息

在 2014 年 3 月 23 日，畿尼的健康的部通知了在东南的 forested 区域的 Ebola 病毒疾病(EVD ) 的很快传播的爆发，并且 4 ? 天以后，疾病被证实被传给 Conakry，畿尼的首都。疾病很快也传播了到利比里亚，尼日利亚和塞拉利昂。2014 年 7 月 27 日，累积数字证实(909 ) ，可能(276 ) 并且在上述四个国家的嫌疑犯(138 ) EVD 盒子到达了 1,323，包括 729 死亡(与 55.1 的死亡率 ?%)(http://***/csr/don/2014_07_31_ebola/en/) 。这是 EVD 的最大的爆发直到现在并且第一次疾病在韦斯特检测了非洲。

关键词：埃博拉病毒 EVD 森林地区利比里亚塞拉利昂尼日利亚 HTTP 几内亚

Assessment of the risk of infectious aerosols leaking to the environment from BSL-3 laboratory HEPA air filtration systems using model bacterial aerosols

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Particuology 2014年第2期12卷 82-87页

作者： Zhanbo Wen Wenhui Yang Na Li Jie Wang Lingfei Hu Jinsong Li Zhe Yin Ke Zhang Xiaokai Dong State Key Laboratory of Pathogen and Biosecurity Beijing Institute of Microbiology and Epidemiology

To assess the risk of infectious bacterial aerosols leaking to the environment, the filtration efficiency of a biosafety level 3 （BSL-3） laboratory high-efficiency particulate （HEPA） filter was investigated using the aerosolized bacteria Serratia marcescens. The aerosol size was measured using an Andersen sampler. Eight first stage HEPA filters （numbered 1-87 were distributed in contaminated labs and exhausts from each of the first stage HEPA filters were aggregated and filtered through one second stage HEPA filter before being released to the environment. In total, 8 first-stage and I second-stage HEPA filters from the BSL-3 air purification system were analyzed. No S. marcescens was detected in first stage filters 1,2, 4, 5, 7 and 8 and the second stage HEPA filter. The filtration efficiencies against aerosolized S. marcescens were 〉99.9999%. First stage filter numbers 3 and 6 had filtration efficiencies of 99.9825% and 99.9906% respectively. When filter number 3 was replaced by a new filter and the bracket for filter number 6 was sealed, no aerosolized S. marcescens was detected in the filtered air. Our work suggests that the BSL-3 laboratory HEPA filter air purification system is effective against bacterial aerosols, with little to no bacterial leakage into the environment.

关键词： Bacterial aerosolFiltration efficiencyHEPA filtersSerratia marcescens

A small,polyphyletic group of Firmicutes synthesizes trimethylamine from L-carnitine

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mLife 2023年第3期2卷 267-271页

作者： Marius Vital Ylenia Heinrich-Sanchez Hannover medical School Institute for Medical Microbiology and Hospital EpidemiologyHannoverGermany

Impact statement Gut microbiota-derived trimethylamine(TMA)is associated with cardiometabolic disorders and exemplifies a microbial involvement in the etiology of emerging,noncommunicable diseases,the leading causes of death *** bio-chemical pathways taking dietary compounds as intake have been described with distinct taxa involved that are all present at low relative abundances.A recently discovered pathway is now considered to be the main route for TMA synthesis from L-carnitine involvingγ-butyrobetaine as an intermediate *** comprehensive(meta)genomic screening of publicly available data,namely,genomes of the UHGG catalog(n>200,000)and 10 metagenomic(transcriptomic)data sets,we revealed bacteria synthesizing TMA via this pathway and specified their *** will contribute to stratification of individuals based on their gut microbiota's potential to synthesize TMA and might aid in the development of strategies restricting TMA formation.

关键词： product. synthesize statement

Anti TB drug resistance in Tanga,Tanzania:a cross sectional facility base prevalence among pulmonary TB patients

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Asian Pacific Journal of Tropical Medicine 2015年第11期8卷 887-893页

作者： Ha Abubakar S.Hoza Sayoki G.M.Mfinanga Brigitte Knig Department of medical microbiology & epidemiology of Infectious Diseases Medical FacultyUniversity of Leipzig Department of microbiology & Parasitology.Sokoine University of Agriculture National institute for medical Research(NIMR)

Objective:To determine the prevalence and risk factors associated with drug resistance tuberculosis(TB) at facility-base level in Tanga,***:A total of 79 Mycobacterium tuberculosis(MTB) isolates included in the study were collected from among372(312 new and 60 previously treated) TB suspects self referred to four TB clinics during a prospective study conducted from November 2012 to January *** and drug susceptibility test of the isolates was performed at the institute of medical microbiology and epidemiology of infectious diseases,University hospital,Leipzig,*** on the patient's characteristics were obtained from structured questionnaire administered to the patients who gave informed verbal *** bivariate logistic regression analysis was performed to assess the risk factors for drug *** significance level was determined at P<***:The overall proportions of any drug resistance and MDRTB were 12.7%and 6.3%*** prevalence of any drug resistance and MDRTB among new cases were 11.4%and 4.3%respectively,whereas among previously treated cases was 22.2%*** treated patients were more likely to develop anti-TB drug *** was no association between anti-TB drug resistances(including MDRTB) with the risk factors ***:High proportions of anti TB drug resistance among new and previously treated cases observed in this study suggest that,additional efforts still need to be done in identifying individual cases at facility base level for improved TB control programmes and drug resistance survey should continuously be monitored in the country.

关键词： Pulmonary tuberculosis Prevalence Drug resistance

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World Journal of Gastroenterology 2015年第13期21卷 4006-4013页

作者： Yuan-Yuan Zhang Hong-Bo Chen Yin Xu Peng Huang Jie Wang Yun Zhang Rong-Bin Yu Jing Su Department of epidemiology and Biostatistics School of Public HealthNanjing Medical University Department of Infectious Diseases Jurong Peoples’ Hospital Department of General Practice Kangda CollegeNanjing Medical University institute of epidemiology and microbiology the Institute of Military Medicine of Nanjing Command

AIM: To determine the association between rapid viral response and IL28 B, IL28 RA, IL10 RB and Mx A polymorphisms in the Chinese Han ***: The study cohort consisted of 238 chronic hepatitis C patients treated with interferon(IFN)-α-2b and ribavirin. Six single nucleotide polymorphisms were genotyped using the ABI Taq Man allelic discrimination assay. Biochemical indices were measured at baseline. Serum hepatitis C virus(HCV) RNA was detected at weeks 0, 4, 12 and 24 of ***: Only IL28 B rs12980275 was associated with treatment response in the Chinese Han population. Patients carrying AG/GG genotypes had a reduced rapid viral response compared with patients carrying the AA genotype(additive model: adjusted OR = 0.43, 95%CI: 0.24-0.75). It took less time for patients with the AA genotype to achieve a viral load < 500 copies/m L(logrank test, P = 0.004). In addition, the protective effect of genotype AA was independent of baseline viral load. HCV genotype, and baseline white blood cell count, α-fetoprotein and viral load might also help predict treatment response. The area under the receiveroperating characteristic curve was 0.726. CONCLUSION: IL28 B rs12980275 AA genotype is a strong predictor of positive response to IFN therapy in Chinese Han patients with hepatitis C.

关键词： Hepatitis C virus Interferon Rapid viral response

Isolation and characterization of dengue virus serotype 2 from the large dengue outbreak in Guangdong, China in 2014

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Science China(Life Sciences) 2014年第12期57卷 1149-1155页

作者： ZHAO Hui ZHAO Ling Zhai JIANG Tao LI Xiao Feng FAN Hang HONG Wen Xin ZHANG Yu ZHU Qin YE Qing TONG Yi Gang CAO Wu Chun ZHANG Fu Chun QIN Cheng Feng State Key Laboratory of Pathogen and Biosecurity Beijing Institute of Microbiology and Epidemiology Guangzhou 8th People's hospital Guangzhou Medical University

Dengue has been well recognized as a global public health threat,but only sporadic epidemics and imported cases were reported in recent decades in *** July 2014,an unexpected large dengue outbreak has occurred in Guangdong province,China,resulting in more than 40000 patients including six *** clarify and characterize the causative agent of this outbreak,the acute phase serum from a patient diagnosed with severe dengue was subjected to virus isolation and high-throughput sequencing(HTS).Traditional real-time RT-PCR and HTS with Ion Torrent PGM detected the presence of dengue virus serotype 2(DENV-2).A clinical DENV-2 isolate GZ05/2014 was obtained by culturing the patient serum in mosquito C6/36 *** complete genome of GZ05/2014 was determined and deposited in Gen Bank under the access number *** analysis based on the complete envelope gene showed that the newly DENV-2 isolate belonged to Cosmopolitan genotype and clustered closely with other Guangdong strains isolated in the past *** amino acid mutations that are obviously known to increase virulence or replication were identified throughout the genome of GZ05/*** high homology of Guangdong DENV-2 strains indicated the possibility of establishment of local DENV-2 circulation in Guangdong,*** results help clarify the origin of this epidemic and predict the future status of dengue in China.

关键词： dengue virus serotype 2 virus isolation phylogenetic analysis envelope gene

Infectious diseases during the European Union training mission Mali(EUTM MLI)–a four-year experience

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Military medical Research 2018年第4期5卷 293-303页

作者： Hagen Frickmann Ralf Matthias Hagen Florian Geiselbrechtinger Nagpal Hoysal Department of microbiology and hospital Hygiene Bundeswehr Hospital Hamburg institute for medical microbiology Virology and Hygiene University Medicine Rostock Department of Preventive Medicine Bundeswehr Medical Academy NATO Center of Excellence for Military Medicine (MilMedCOE) Deployment Health Surveillance Capability (DHSC) institute of medical Information Processing Biometry and Epidemiology

Background: The European Union Training Mission Mali(EUTM MLI) is a multinational military training deployment to the Western African tropical nation of Mali. Based on routinely collected disease and non-battle injury surveillance data, this study quantifies the true impact of infectious diseases for this tropical mission and potential seasonal variations in infectious disease ***: Categorized health events during the EUTM MLI mission and associated lost working days were reported using the EpiNATO-2 report. Infection-related health events were descriptively analyzed for a 4-year period from the 12 th week in 2013 to the 13 th week in 2017. Aggregated EpiNATO-2 data collected from all missions other than EUTM MLI were used as a ***: Among the infectious diseases reported by EUTM MLI, non-severe upper respiratory infections and gastrointestinal diseases dominated quantitatively, accounting for 1.65 and 1.42 consultations per 100 person-weeks, respectively. The number of recorded infectious disease-associated lost working days during the whole study interval was 723. Seasonal changes in disease frequency were detectable. More gastrointestinal infections were seen in the rainy season, and more respiratory infections occurred in the dry season; these were associated with peaks of more than 2.5 consultations per 100 person-weeks for both ***: Despite initial concerns focused on tropical infectious diseases during this mission in tropical Mali, upper respiratory tract and gastrointestinal infections predominate. The relatively low number of reported lost working days may indicate that these infections are at the milder end of the spectrum of infectious diseases despite a likely reporting bias.

关键词： Tropical deployment Infectious diseases Tropical medicine Gastrointestinal infections Upper respiratory tract infections Mali