Comparison of immune responses and intestinal flora in epicutaneously sensitized BALB/c or C57BL/6 mouse models of food allergy

Cutaneous exposure to food allergens through a disrupted skin barrier is recognized as an important cause of food allergy,and the cutaneous sensitized mouse model has been established to investigate relevant allergic disorders.However,the role of different genetic backgrounds of mice on immune responses to food allergens upon epicutaneous sensitization is largely unknown.In this study,two strains of mice,i.e.,the BALB/c and C57BL/6 mice,were epicutaneously sensitized with ovalbumin on atopic dermatitis(AD)-like skin lesions,followed by intragastric challenge to induce IgE-mediated food allergy.Allergic outcomes were measured as clinical signs,specific antibodies and cytokines,and immune cell subpopulations,as well as changes in intestinal barrier function and gut microbiota.Results showed that both strains of mice exhibited typical food-allergic symptoms with a Th2-skewed response.The C57BL/6 mice,rather than the BALB/c mice,were fitter for establishing an epicutaneously sensitized model of food allergy since a stronger Th2-biased response and severer disruptions in the intestinal barrier and gut homeostasis were observed.This study provides knowledge for selecting an appropriate mouse model to study food-allergic responses associated with AD-like skin lesions and highlights the role of genetic variations in the immune mechanism underlying pathogenesis of food allergy.

Comparative Study on the Immunogenicity and Efficacy of Different Post-exposure Intramuscular Rabies Vaccination Regimens in China

Objective This study aimed to compare the current Essen rabies post-exposure immunization schedule(0-3-7-14-28)in China and the simple 4-dose schedule(0-3-7-14)newly recommended by the World Health Organization in terms of their safety,efficacy,and protection.Methods Mice were vaccinated according to different immunization schedules,and blood was collected for detection of rabies virus neutralizing antibodies(RVNAs)on days 14,21,28,35,and 120after the first immunization.Additionally,different groups of mice were injected with lethal doses of the CVS-11 virus on day 0,subjected to different rabies immunization schedules,and assessed for morbidity and death status.In a clinical trial,185 rabies-exposed individuals were selected for post-exposure vaccination according to the Essen schedule,and blood was collected for RVNAs detection on days 28and 42 after the first immunization.Results A statistically significant difference in RVNAs between mice in the Essen and 0-3-7-14 schedule groups was observed on the 35th day(P<0.05).The groups 0-3-7-14,0-3-7-21,and 0-3-7-28 showed no statistically significant difference(P>0.05)in RVNAs levels at any time point.The post-exposure immune protective test showed that the survival rate of mice in the control group was 20%,whereas that in the immunization groups was 40%.In the clinical trial,the RVNAs positive conversion rates on days 28(14 days after 4 doses)and 42(14 days after 5 doses)were both 100%,and no significant difference in RVNAs levels was observed(P>0.05).Conclusion The simple 4-dose schedule can produce sufficient RVNAs levels,with no significant effect of a delayed fourth vaccine dose(14–28 d)on the immunization potential.

Rapid identification of full-length genome and tracing variations of monkeypox virus in clinical specimens based on mNGS and amplicon sequencing

The monkeypox virus(MPXV)has triggered a current outbreak globally.Genome sequencing of MPXV and rapid tracing of genetic variants will benefit disease diagnosis and control.It is a significant challenge but necessary to optimize the strategy and application of rapid full-length genome identification and to track variations of MPXV in clinical specimens with low viral loads,as it is one of the DNA viruses with the largest genome and the most AT-biased,and has a significant number of tandem repeats.Here we evaluated the performance of metagenomic and amplicon sequencing techniques,and three sequencing platforms in MPXV genome sequencing based on multiple clinical specimens of five mpox cases in Chinese mainland.We rapidly identified the full-length genome of MPXV with the assembly of accurate tandem repeats in multiple clinical specimens.Amplicon sequencing enables cost-effective and rapid sequencing of clinical specimens to obtain high-quality MPXV genomes.Third-generation sequencing facilitates the assembly of the terminal tandem repeat regions in the monkeypox virus genome and corrects a common misassembly in published sequences.Besides,several intra-host single nucleotide variations were identified in the first imported mpox case.This study offers an evaluation of various strategies aimed at identifying the complete genome of MPXV in clinical specimens.The findings of this study will significantly enhance the surveillance of MPXV.

Viral and Bacterial Etiology of Acute Febrile Respiratory Syndrome among Patients in Qinghai, China

Objective This study was conducted to investigate the viral and bacterial etiology and epidemiology of patients with acute febrile respiratory syndrome(AFRS) in Qinghai using a commercial routine multiplex-ligation-nucleic acid amplification test(NAT)-based assay. Methods A total of 445 nasopharyngeal swabs specimens from patients with AFRS were analyzed using the RespiFinderSmart22 kit(PathoFinder BV, Netherlands) and the LightCycler 480 real-time PCR system. Results Among the 225(225/445, 51%) positive specimens, 329 positive pathogens were detected, including 298(90.58%) viruses and 31(9%) bacteria. The most commonly detected pathogens were influenza virus(IFV;37.39%;123/329), adenovirus(AdV;17.02%;56/329), human coronaviruses(HCoVs;10.94%;36/329), rhinovirus/enterovirus(RV/EV;10.03%;33/329), parainfluenza viruses(PIVs;8.51%;28/329), and Mycoplasma pneumoniae(M. pneu;8.51%;28/329), respectively. Among the co-infected cases(17.53%;78/445), IFV/AdV and IFV/M. pneu were the most common co-infections. Most of the respiratory viruses were detected in summer and fall. Conclusion In our study, IFV-A was the most common respiratory pathogen among 22 detected pathogens, followed by AdV, HCoV, RV/EV, PIV, and M. pneu. Bacteria appeared less frequently than viruses, and co-infection was the most common phenomenon among viral pathogens. Pathogens were distributed among different age groups and respiratory viruses were generally active in July, September, and November. Enhanced surveillance and early detection can be useful in the diagnosis, treatment, and prevention of AFRS, as well as for guiding the development of appropriate public health strategies.

A Reverse-transcription Recombinase-aided Amplification Assay for the Rapid Detection of the Far-Eastern Subtype of Tick-borne Encephalitis Virus

Objective Tick-borne encephalitis virus(TBEV) is an emerging pathogen in Europe and North Asia that causes tick-borne encephalitis(TBE). A simple, rapid method for detecting TBEV RNA is needed to control this disease. Methods A reverse-transcription recombinase-aided amplification(RT-RAA) assay was developed. This assay can be completed in one closed tube at 39℃ within 30 minutes. The sensitivity and specificity of RT-RAA were validated using non-infectious synthetic RNA representing a fragment of the NS5 region of the wild-type(WT) TBEV genome and the Senzhang strain. Additionally, 10 batches of tick samples were used to evaluate the performance of the RT-RAA assay. Results The analytical limit of detection of the assay was 20 copies per reaction of the TBEV synthetic transcript and 3 plaque-forming units(pfu) per reaction of TBEV titers. With the specific assay, no signal due to other arboviruses was observed. Of the 10 batches of tick samples obtained from the Changbai Mountains of China, three were TBEV-positive, which was consistent with the results of the quantitative real-time PCR assay. Conclusion A rapid, highly sensitive, specific, and easy-to-use method was developed for the detection of the TBEV Far-Eastern subtype.

Detection of Pseudorabies Virus Antibodies in Human Encephalitis Cases

Pseudorabies virus(PRV),a veterinary pathogen that infects domestic animals as well as wild animals such as wild boar and feral swine,was recently reported to infect human and led to endophthalmitis and encephalitis.A retrospective seroepidemiologic survey was conducted using 1,335 serum samples collected from patients with encephalitis and ELISA positive rates were 12.16%,14.25%,and 6.52%in 2012,2013,and 2017,respectively.

Age-related rhesus macaque models of COVID-19

Background:Since December 2019,an outbreak of the Corona Virus disease 2019(COVID-19)caused by severe acute respiratory syndrome coronavirus(SARS-CoV-2)in Wuhan,China,has become a public health emergency of international concern.The high fatality of aged cases caused by SARS-CoV-2 was a need to explore the possible age-related phenomena with non-human primate models.Methods:Three 3-5 years old and two 15 years old rhesus macaques were intratracheally infected with SARS-CoV-2,and then analyzed by clinical signs,viral replication,chest X-ray,histopathological changes and immune response.Results:Viral replication of nasopharyngeal swabs,anal swabs and lung in old monkeys was more active than that in young monkeys for 14 days after SARS-CoV-2 challenge.Monkeys developed typical interstitial pneumonia characterized by thickened alveolar septum accompanied with inflammation and edema,notably,old monkeys exhibited diffuse severe interstitial pneumonia.Viral antigens were detected mainly in alveolar epithelial cells and macrophages.Conclusion:SARS-CoV-2 caused more severe interstitial pneumonia in old monkeys than that in young monkeys.Rhesus macaque models infected with SARS-CoV-2 provided insight into the pathogenic mechanism and facilitated the development of vaccines and therapeutics against SARS-CoV-2 infection.

Development of an Internally Controlled Reverse Transcription Recombinase-aided Amplification Assay for the Rapid and Visual Detection of West Nile Virus

West Nile virus(WNV)causes West Nile fever and West Nile encephalitis.Because infection by WNV creates serious public health problems,its simple,rapid,and visual detection is very important in clinical practice,especially in resource-limited laboratories.We have developed a rapid,specific,and highly sensitive internally controlled reverse transcription recombinase-aided amplification(RTRAA)assay to detect WNV,using both real-timefluoresce nee and the lateral flow dipstick(LFD)at39.0°C for 30 min.The analytical sensitivity of theRT-RAA assay was 10 plasmid copies and 1.6 pfu perreacti on with real-time fluoresce nee,and 1,000plasmid copies per reaction with the LFD.No crossreactionwith other control viruses was observed.Compared with the RT-qPCR assay,the RT-RAA assaydemonstrated 100%sensitivity and 100%specificityfor WNV.

Molecular Characterisation of Two Coxsackievirus B6 Strains from the Tibet Autonomous Region of China

Enteroviruses (EVs) are members of the genus Enteroviruswithin the orderPicornavirales, family Picornaviridae, and consist of 12 species, including EV-A, EV-B, EV-C, and EV-D, which are associated with human infections. Coxsackievirus B6 (CV-B6)belongs to the species EV-B, which currently consists of 63 serotypes, including echovirus (serotypes 1–7,9, 11–21, 24–27, 29–33), coxsackievirus group A (CVA9), coxsackievirus group B (CV-B, serotypes 1–6),the newly identified EVs (serotypes EV-B69, B73–75,B77–88, B93, B97–98, B100–101, B106–107, and B110–113), and the simian enterovirus SA5.

Japanese Encephalitis in China in the Period of 1950–2018:From Discovery to Control

Japanese encephalitis(JE)was first discovered in Japan in 1871;in 1924,a major outbreak occurred,with 6,000 JE cases reported and a mortality rate of approximately 60%[1,2].Later studies showed that JE is caused by the Japanese encephalitis virus(JEV),which is spread by mosquitoes.

Serological Survey of Zika Virus in Humans and Animals in Dejiang Prefecture, Guizhou Province,China

Objective The current outbreak of Zika virus(ZIKV)poses a severe threat to human health.Two ZIKV strains were isolated from mosquitoes collected from the Dejiang prefecture in China in 2016,which was the first isolation of ZIKV in nature in China.Methods In this study,serum samples were collected from 366 healthy individuals and 104 animals from Dejiang prefecture in 2017,and the plaque reduction neutralization test(PRNT)was used to evaluate the seroprevalence of ZIKV.Results None of the 366 residents from whom the samples were collected were seropositive for ZIKV.None of the 11 pigs from whom the samples were collected were seropositive for ZIKV,while 1 of 63(1.59%)chickens and 2 of 30(6.67%)sheep were seropositive for ZIKV.Conclusions The extremely low seropositivity rate of ZIKV antibodies in animals in the Dejiang prefecture,Guizhou province in this study indicates that ZIKV can infect animals;however,there is a low risk of ZIKV circulating in the local population.

Estimation of evaporation losses based on stable isotopes of stream water in a mountain watershed

Water stable isotopes(δ^(2) H andδ^(18)O)can record surface water evaporation,which is an important hydrological process for understanding watershed structure and function evolution.However,the isotopic estimation of water evaporation losses in the mountain watersheds remains poorly explored,which hinders understanding spatial variations of hydrological processes and their relationships with the temperature and vegetation.Here we investigatedδ^(2) H,δ^(18)O,and d-excess values of stream water along an altitude gradient of 2130 to 3380 m in Guan’egou mountain watershed at the east edge of the Qinghai-Tibet Plateau in China.The meanδ^(2) H(-69.6‰±2.6‰),δ^(18)O(-10.7‰±0.3‰),and dexcess values(16.0‰±1.4‰)of stream water indicate the inland moisture as the major source of precipitation in study area.Water stable isotopes increase linearly with decreasing altitudes,based on which we estimated the fractions of water evaporation losses along with the altitude and their variations in different vegetations.This study provides an isotopic evaluation method of water evaporation status in mountain watersheds,the results are useful for further understanding the relationship between hydrological processes and ecosystem function under the changing climate surrounding the Qinghai-Tibet Plateau.

A New Species of Trimeresurus Lacépède, 1804(Squamata: Viperidae) from Southwestern China, Vietnam, Thailand and Myanmar

The pit vipers of the genus Trimeresurus Lacépède, 1804 is one of the largest groups of Asian snakes, distributed from India to China and Indonesia. Recent surveys in Jiangcheng and Simao, Yunnan Province, China resulted in a new species previously allocated to T. albolabris. Combining morphological and molecular data, we describe it as Trimeresurus guoi sp. nov. The new species morphologically differs from T. albolabris in the yellow green ventral color;an indistinct ventrolateral line;the a bsence of a postocular stripe;the firebrickred iris;a dark red stripe on dorsal tail;hemipenes with relatively weak sparse papillae, reaching 23 rd subcaudal when unextruded. Molecularly, the new species forms a clearly divergent lineage(BPP 1.00/UFB 100). Uncorrected pairwise distances of mitochondrial gene Cyt b between the new species and other known species of the subgenus Trimeresurus range from 0.052(T. albolabris) to 0.071(T. insularis).

A Reverse-Transcription Recombinase-Aided Amplification Assay for the Rapid Detection of the Wuxiang Virus

Wuxiang virus(WUXV),a new species of Phlebovirus from sandfly specimens(identified as Phlebotomus chinensis)collected in Wuxiang County,Shanxi Province,China,belongs to the order Bunyavirales,family Phenuiviridae.Like other Bunyavirales,WUXV contains a negative-sense tripartite RNA genome comprising a small(S),a medium(M),and a large(L)segment.The S segment encodes nucleocapsid protein(NP)and a nonstructural protein(NSP).The M segment encodes a glycoprotein precursor(Gp),which is cleaved into mature N and C glycoproteins.The L segment encodes an RNA-dependent RNA polymerase(RdRp).

Perspective on the application of genome sequencing for monkeypox virus surveillance

Monkeypox virus(MPXV),a DNA virus belonging to the Orthopoxvirus genus,causes a self-limiting zoonotic disease known as mpox.The human monkeypox infection was first recorded in a 9-month-old child in the Republic of Congo in the 1970s(Ladnyj et al.,1972).For a long time,mpox was mainly prevalent in the humid forests of Central Africa and parts of West Africa(Kabugae and El Zowalaty,2019).In recent years,MPXV has been spread to other countries through trade and travel.In 2003,a group of rare pets carrying MPXV was exported from Africa to the United States,and an outbreak of animal-to-human transmission occurred(Di Giulio and Eckburg,2004).In 2018,two cases of mpox were confirmed in travelers from Nigeria to the United Kingdom and one case was confirmed in Israel(Vaughan et al.,2018;Erez et al.,2019).Then,in 2019,one case was confirmed in Singapore(Ng et al.,2019).The outbreak of mpox in multiple non-endemic countries in North America and Europe started in May 2022(WHO,2022b).On July 23,2022,the WHO declared MPXV a Public Health Emergency of International Concern(PHEIC)(WHO,2022a).As of March 5,2023,86,309 confirmed mpox cases have been reported from 107 countries/regions worldwide(WHO,2023).In the mainland of China,the first imported case was confirmed by the Chinese Center for Disease Control and Prevention(Zhao et al.,2022),making this the fifth confirmed mpox infection in China.Neglected zoonotic mpox has been restricted in Africa but now it is back in the spotlight worldwide(Tan and Gao,2022).

Characterization of whole genomes from recently emerging Mpox cases in several regions of China,2023

Dear Editor,Mpox(formerly known as monkeypox)is a zoonotic disease caused by infection with the monkeypox virus(MPXV).Mpox cases have been sporadic over the past few decades,with outbreaks occurring in only a limited number of countries,primarily as a result of imported cases(El Eid et al.,2022;Tan and Gao,2022).

Mpox(formerly monkeypox):pathogenesis,prevention,and treatment

In 2022,a global outbreak of Mpox(formerly monkeypox)occurred in various countries across Europe and America and rapidly spread to more than 100 countries and regions.The World Health Organization declared the outbreak to be a public health emergency of international concern due to the rapid spread of the Mpox virus.Consequently,nations intensified their efforts to explore treatment strategies aimed at combating the infection and its dissemination.Nevertheless,the available therapeutic options for Mpox virus infection remain limited.So far,only a few numbers of antiviral compounds have been approved by regulatory authorities.Given the high mutability of the Mpox virus,certain mutant strains have shown resistance to existing pharmaceutical interventions.This highlights the urgent need to develop novel antiviral drugs that can combat both drug resistance and the potential threat of bioterrorism.Currently,there is a lack of comprehensive literature on the pathophysiology and treatment of Mpox.To address this issue,we conducted a review covering the physiological and pathological processes of Mpox infection,summarizing the latest progress of anti-Mpox drugs.Our analysis encompasses approved drugs currently employed in clinical settings,as well as newly identified small-molecule compounds and antibody drugs displaying potential antiviral efficacy against Mpox.Furthermore,we have gained valuable insights from the process of Mpox drug development,including strategies for repurposing drugs,the discovery of drug targets driven by artificial intelligence,and preclinical drug development.The purpose of this review is to provide readers with a comprehensive overview of the current knowledge on Mpox.

Efficient inhibition of SARS-CoV-2 emerging EG.5,EG.5.1 and BA.2.86 variants by fusion inhibitor HY3000 peptide

Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)continues to pose a significant threat to the world,as it continually evolves and gives rise to multiple variants and sub-variants.Recently,the Omicron EG.5 linage,which was first detected in Indonesia on 17 February 2023,has raised concerns due to its increased prevalence and extended immune escape properties,according to the risk analysis by the World Health Organization(WHO)[1].As of 3 October 2023,EG.5 and its sub-linages have been reported in 83 countries with shared 49,008 genome sequences in GISAID database(https://gisaid.org/hcov19-variants/).EG.5 has become the dominant strain in the United States according to Centers for Disease Control and Prevention(CDC),accounting for 29.4%of SARS-CoV-2 infections(https://covid.cdc.gov/covid-data-tracker/#variant-proportions).

Establishment of a humanized ST6GAL1 mouse model for influenza research

Background:This study aimed to construct and characterize a humanized influenza mouse model expressing hST6GAL1.Methods:Humanized fragments,consisting of the endothelial cell-specific K18 promoter,human ST6GAL1-encoding gene,and luciferase gene,were microinjected into the fertilized eggs of mice.The manipulated embryos were transferred into the oviducts of pseudopregnant female mice.The offspring were identified using PCR.Mice exhibiting elevated expression of the hST6GAL1 gene were selectively bred for propagation,and in vivo analysis was performed for screening.Expression of the humanized gene was tested by performing immunohistochemical(IHC)analysis.Hematologic and biochemical analyses using the whole blood and serum of humanized hST6GAL1 mice were performed.Results:Successful integration of the human ST6GAL1 gene into the mouse genome led to the overexpression of human SiaT ST6GAL1.Seven mice were identified as carrying copies of the humanized gene,and the in vivo analysis indicated that hST6GAL1gene expression in positive mice mirrored influenza virus infection characteristics.The IHC results revealed that hST6GAL1 was expressed in the lungs of humanized mice.Moreover,the hematologic and biochemical parameters of the positive mice were within the normal range.Conclusion:A humanized influenza mouse model expressing the hST6GAL1 gene was successfully established and characterized.

Development of two multiplex real-time PCR assays for simultaneous detection and differentiation of monkeypox virus IIa,IIb,and I clades and the B.1 lineage

An ongoing multi-country outbreak of monkeypox was reported in May 2022 with several deaths,affecting 107 countries of all six World Health Organization(WHO)regions.The WHO has declared the current monkeypox outbreak to be a Public Health Emergency of International Concern.It is,thus,necessary to rapidly and accurately detect and distinguish different monkeypox virus(MPXV)clades.We designed primers and probes based on the alignment of 138 complete genomes of poxviruses.In Panel 1,we mixed one pair of primers and three probes to detect and differentiate the MPXV Western Africa(IIa,IIb clade)and Congo Basin(I clade)and other orthopoxviruses.In Panel 2,we mixed one pair of primers and two probes to detect the 2022 MPXV(B.1 lineage and its descendant lineages).In addition,we tested the specificity and sensitivity of the assay using real-time PCR.In Panel 1,the assay reproducibly identified various concentrations of two plasmids of the monkeypox virus,whereas other orthopoxviruses did not cross-react.In Panel 2,the probe annealed well to MPXV B.1 and showed the expected linearity.These two multiple real-time assays are inclusive and highly specific for identifying different clades of MPXV.