The Taxonomic Status of Spermophilus in the Plague Area of Dingbian County,Shaanxi Province,China

This study was conducted to define the taxonomic status of Spermophilus in the plague area of Dingbian County in Shaanxi Province,China,through the two-factor variance analysis of morphological characteristics,DNA barcoding,and chromosome karyotype analysis.

Bartonella Species Investigated among Rodents from Shaanxi Province of China

Bartonella spp.are rod-shaped,Gram-negative,aerobic,fastidious,slow bacteria which cause diseases in humans and animals by parasitizing the endothelial and red blood cells of their hosts[1,2].Rodents are the most important natural reservoir hosts of Bartonella[3].In 57 countries,epidemiological studies of rat-borne Bartonella have been carried out.Information on the Bartonella infection rate in rodents has been obtained from most of these countries.The infection rate of rodents in Portugal,Egypt,Japan,Canada,and the United States is more than 90%,the infection rate in Thailand and Russia is about 80%,and the infection rate in China is about 67%.To date,22 species of rodent-borne Bartonella have been described.Of these,B.grahamii,B.vinsonii subsp.arupensis,B.vinsonii subsp.Berkhoffii,and B.elizabethae have been found to be associated with human illness.People become infected with rodent-borne Bartonella incidentally,especially when they are exposed to the habitats of wild rodents harboring various Bartonella species.Bartonella infection can affect multiple organs and poses a risk to public health[4].

Construction of recombinant adeno-associated virus vector co-expressing hVEGF_(165) and hBMP_7 gene

Objective： To construct recombinant adeno-associated virus co-expressing human vascular epithelial growth factor 165（hVEGF165） and bone morphogenetic protein 7（hBMPT）, measure the virus titer and verify the recombination. Methods：The AAV helper-free system was used as basis to generate recombinant AAV. The IRES sequence of plasmid plRES was cut down and subcloned into ITR/ MCS containing vector pAAV-MCS to construct recombinant plasmid pAAV-MCSa-IRES-MCSb. The hVEGF165 and hBMP7 gene was amplified by PCR and inserted into upstream MCSa and downstream MCSb respectively. Then, recombinant plasmid pAAV- hVEGF165-IRES-hBMP7, pAAV-RC and pHelper were co-transfected into AAV-293 cells to complete rAAV-hVEGF165-IRES-hBMP7 packaging. The GFP labeled rAAV-IRES-GFP was simultaneously packaged by using the parallel plasmid pAAV-IRES-hrGFP. The efficiency of AAV packaging was monitored under fluorescent microscope and recombinant viral particles were harvested from infected AAV-293 cells. The virus titer was measured by infecting AAV-HT1080 cells, and the recombinant AAV-hVEGF165-IRES-hBMP7 was verified by PCR of the exogenous interest genes. Results：Recombinant pAAV-hVEGF165-IRES-hBMP7 was verified by double digestion. GFP expression in AAV-293 could be observed under fluorescent microscope 72 h after transfection and the system provided a high packing ratio of 95%. The recombinant adeno-associated virus has a high titer of 5.5 ×10^11vp/ml, and AAV-HT 1080 was infected at a ratio of 90%. The recombinant virus was confirmed by PCR of exogenous hBMP7 and hVEGF165 gene. Conclusion：Recombinant rAAV-hVEGF165-IRES-hBMP7 was successfully constructed with a high virus titer, which may offer foundation for in vitro and in vivo experiments of hVEGF165 and hBMP7 co-expression and provide a new method for gene therapy of bone regeneration.

Novel astrovirus and paramyxovirus in Mongolian gerbils(Meriones unguiculatus)from China

DEAR EDITOR,Mongolian gerbils(Meriones unguiculatus)are small rodents belonging to the family Muridae and widely inhabit typical steppes and desert grasslands across northern China.Yet,few viruses in wild M.unguiculatus have been reported.Using meta-transcriptomic sequencing we identified two novel RNA viruses:an astrovirus,denoted Gerbil astrovirus(GeAstV).

Evolutionary Diversity of Coxsackievirus A6 Causing Severe Hand,Foot,and Mouth Disease—China,2012-2023

Introduction:Coxsackievirus A6(CVA6)has emerged as a significant pathogen responsible for severe cases of hand,foot,and mouth disease(HFMD).This study aims to delineate the demographic characteristics and analyze the viral evolution of severe HFMD associated with CVA6,thereby assisting in its surveillance and management.Methods:In this investigation,74 strains of CVA6 were isolated from samples collected from severe HFMD cases between 2012 and 2023.The VP1 gene sequences of CVA6 were amplified and analyzed to assess population historical dynamics and evolutionary characteristics using BEAST,DnaSP6,and PopART.Results:A significant portion(94.4%)of severe CVA6-associated HFMD cases(51 out of 54,with 20 lacking age information)were children under 5 years old.Among the 74 CVA6 strains analyzed,72 belonged to the D3a sub-genotype,while only two strains were D2 sub-genotype.The average genetic distance between VP1 sequences prior to 2015 was 0.027,which increased to 0.051 when compared to sequences post-2015.Historical population dynamics analysis indicated three significant population expansions of severe CVA6-associated HFMD during 2012-2013,2013-2014,and 2019-2020,resulting in the formation of 65 distinct haplotypes.Consistent with the MCC tree findings,transitioning between regional haplotypes required multiple base substitutions,showcasing an increase in population diversity during the evolutionary process(from 14 haplotypes in 2013 to 55 haplotypes over the subsequent decade).Conclusions:CVA6,associated with severe HFMD,is evolving and presents a risk of outbreak occurrence.Thus,enhanced surveillance of severe HFMD is imperative.

Establishment of a recombinant adeno-associated virus expressing hVEGF_(165)

BACKGROUND： Because certain gene vectors could have deleterious effects in the central nervous system, the choice of a safe and effective vector system has become more important for gene therapy of nerve regeneration. OBJECTIVE： To construct a non-pathogenic, recombinant adeno-associated virus （AAV） simultaneously expressing human vascular endothelial growth factor 165 （hVEGF165） and green fluorescent protein （GFP）. DESIGN, TIME AND SETTING： A randomized controlled experiment was performed at the Virology Laboratory of Shaanxi Provincial Center for Disease Control and Prevention between March and September 2007. MATERIALS： AAV helper-free system, AAV-293 packaging cell line, and AAV HT-1080 cells were purchased from Stratagene, USA. E. coli DH5α was a stocked strain from Centers for Disease Control and Prevention of Shaanxi, China. Plasmid pUC18-hHVEGF165 was a gift from Zhibin Shi. METHODS： The hVEGF165 gene was amplified by PCR from pUC18-hHVEGF165 and inserted into plasmid pAAV-IRES-hrGFP to construct recombinant plasmid pAAV-hVEGF165-IRES-hrGFP. Subsequently pAAV-hVEGF165-IRES-hrGFP, pAAV-RC （the rep/cap-gene containing plasmid）, and pHelper were co-transfected into AAV-293 cells to complete rAAV-hVEGF165-IRES-hrGFP packaging through homologous recombination. The efficiency of AAV packaging was monitored under a fluorescent microscope, and the recombinant viral particles were harvested from infected AAV-293 cells, and further concentrated and purified. AAV HT-1080 cells were infected with the recombinant virus AAV-hVEGF165-IRES-hrGFP. MAIN OUTCOME MEASURES： Recombinant virus titer was measured by fluorescent cell counting, and infection efficiency was detected by Fluorescence Activated Cell Sorter （FACS） upon infecting AAV-HT1080 cells. The recombination with the exogenous gene was verified by PCR. RESULTS： The PCR amplified products were verified as hVEGF165 gene by DNA sequencing, and the recombinant pAAV-hVEGF165-IRES-GFP was confirmed by double digestion. The system provided a high packaging ratio of 95%, and the purified recombinant virus had a high titer of 5.5×1011 virus particles/mL. The AAV-HT1080 cells were infected at a ratio of 90.4%. The recombinant virus was confirmed by PCR to contain the exogenous hVEGF165 gene. CONCLUSION： The non-pathogenic rAAV-hVEGF165-GFP vector, carrying hVEGF165 and GFP reporter gene, was successfully constructed with a high titer and infection efficiency.

An efficient metatranscriptomic approach for capturing RNA virome and its application to SARS-CoV-2

Coronavirus disease 2019(COVID-19)is a new type of respiratory disease caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),which has caused severe social and economic costs and has become a great threat to public health because of the high mortality rate.Recent studies indicated that respiratory pathogens co-infection and secondary infection were critical risk factors for the severity and mortality rates of COVID-19(Zhou et al.,2020).

Analysis of Epidemiological and Issues Encountered in Case Reports on Echinococcosis—China,2022

Summary What is already known about this topic?Echinococcosis is classified as a Class C infectious disease in China.It is endemic in 370 counties located in the agricultural and pastoral regions of western China.What is added by this report?This report provides a comprehensive overview of the cases of echinococcosis reported in China in 2022.Following a thorough evaluation conducted by provincial CDCs.

The resurgent landscape of xenotransplantation of pig organs in nonhuman primates

Organ shortage is a major bottleneck in allotransplantation and causes many wait-listed patients to die or become too sick for transplantation.Genetically engineered pigs have been discussed as a potential alternative to allogeneic donor organs.Although xenotransplantation of pig-derived organs in nonhuman primates(NHPs)has shown sequential advances in recent years,there are still underlying problems that need to be completely addressed before clinical applications,including(i)acute humoral xenograft rejection;(ii)acute cellular rejection;(iii)dysregulation of coagulation and inflammation;(iv)physiological incompatibility;and(v)cross-species infection.Moreover,various genetic modifications to the pig donor need to be fully characterized,with the aim of identifying the ideal transgene combination for upcoming clinical trials.In addition,suitable pretransplant screening methods need to be confirmed for optimal donor-recipient matching,ensuring a good outcome from xenotransplantation.Herein,we summarize the understanding of organ xenotransplantation in pigs-to-NHPs and highlight the current status and recent progress in extending the survival time of pig xenografts and recipients.We also discuss practical strategies for overcoming the obstacles to xenotransplantation mentioned above to further advance transplantation of pig organs in the clinic.

Genetic source tracking of an anthrax outbreak in Shaanxi province,China

Background:Anthrax is an acute zoonotic infectious disease caused by the bacterium known as Bacillus anthracis.From 26 July to 8 August 2015,an outbreak with 20 suspected cutaneous anthrax cases was reported in Ganquan County,Shaanxi province in China.The genetic source tracking analysis of the anthrax outbreak was performed by molecular epidemiological methods in this study.Methods:Three molecular typing methods,namely canonical single nucleotide polymorphisms(canSNP),multiple-locus variable-number tandem repeat analysis(MLVA),and single nucleotide repeat(SNR)analysis,were used to investigate the possible source of transmission and identify the genetic relationship among the strains isolated from human cases and diseased animals during the outbreak.Results:Five strains isolated from diseased mules were clustered together with patients’isolates using canSNP typing and MLVA.The causative B.anthracis lineages in this outbreak belonged to the A.Br.001/002 canSNP subgroup and the MLVA15-31 genotype(the 31 genotype in MLVA15 scheme).Because nine isolates from another four provinces in China were clustered together with outbreak-related strains by the canSNP(A.Br.001/002 subgroup)and MLVA15 method(MLVA15-31 genotype),still another SNR analysis(CL10,CL12,CL33,and CL35)was used to source track the outbreak,and the results suggesting that these patients in the anthrax outbreak were probably infected by the same pathogen clone.Conclusions:It was deduced that the anthrax outbreak occurred in Shaanxi province,China in 2015 was a local occurrence.

Identification of a novel hepacivirus in Mongolian gerbil(Meriones unguiculatus)from Shaanxi,China

Dear Editor,Hepaciviruses,members of the family Flaviviridae,are enveloped viruses containing a single-stranded positive-sense RNA genome of approximately 8.9-10.5 kb in size(Simmonds et al.,2017).To date,15species(Hepacivirus A-N,and P)have been documented within the Hepacivirus genus that show distinct host ranges,including primates,bats,horses,donkeys,cows,and various rodents(Hartlage et al.,2016).

Epidemiological Data of Work-Related Musculoskeletal Disorders—China,2018–2020

Summary What is already known about this topic?In recent decades,work-related musculoskeletal disorders(WMSDs)have become increasingly prominent and have become an important issue that is of universal concern and an urgent need to be solved in all countries of the world.What is added by this report?The top three industries or occupational groups with the highest standardized prevalence rate of WMSDs were flight attendants,medical staff,and vegetable greenhouses in that order.Women workers were 1.5 times more likely to suffer from WMSDs than men workers.

National Alveolar Echinococcosis Distribution--China, 2012−2016

What is already known about this topic?Both alveolar echinococcosis(AE)and cystic echinococcosis are endemic in China,among which alveolar echinococcosis has a very high mortality rate.What is added by this report?The survey results showed the prevalence and scope of AE in China and identified high-risk groups including children,monks,herdsmen and illiterate people.At the same time,all the cases found in the survey(more than 90%of the patients did not go to the hospital for diagnosis and treatment before survey)were promptly diagnosed and treated.What are the implications for public health practice?This study provides information for the development of a plan for AE prevention and control and for the implementation of interventions targeted to high-risk populations.

What Is Required to Prevent a Second Major Outbreak of SARS-CoV-2 upon Lifting Quarantine in Wuhan City,China

BACKGROUND The Chinese government implemented a metropolitan-wide quarantine of Wuhan city on 23rd January 2020 to curb the epidemic of the coronavirus COVID-19.Lifting of this quarantine is imminent.We modelled the effects of two key health interventions on the epidemic when the quarantine is lifted.