Effects of Differential First-Line Antiretroviral Therapy(ART)Regimens on Mortality among HIV/AIDS Children in Southwest China:A 15-year Retrospective Cohort Study

Human immunodeficiency virus(HIV)is a serious public health concern.Globally,1.7 million(1.3–2.1 million)children aged 0–14 were living with HIV at the end of 2021,90%of whom lived in low-and middle-income countries.Acquired Immune Deficiency Syndrome(AIDS)remains the leading cause of mortality among children worldwide.

Evaluation of Vaporized Hydrogen Peroxide Fumigation as a Method for the Bio-decontamination of the High Efficiency Particulate Air Filter Unit

Objective To evaluate the performance of vaporized hydrogen peroxide （VHP） for the bio-decontamination of the high efficiency particulate air （HEPA） filter unit. Methods Self-made or commercially available bioindicators were placed at designated locations in the HEPA filter unit under VHP fumigation. The spores on coupons were then extracted by 0.5 h submergence in eluent followed by 200- time violent knocks. Results Due to the presence of HEPA filter in the box, spore recovery from coupons placed at the bottom of the filter downstream was significantly higher than that from coupons placed at the other locations. The gap of decontamination efficiency between the top and the bottom of the filter downstream became narrower with the exposure time extended. The decontamination efficiency of the bottom of the filter downstream only improved gently with the injection rate of H202 increased and the decontamination efficiency decreased instead when the injection rate exceeded 2.5 g/min. The commercially available bioindicators were competent to indicate the disinfection efficiency of VHP for the HEPA filter unit. Conclusion The HEPA filter unit is more difficult than common enclosure to decontaminate using VHP fumigation. Complete decontamination can be achieved by extending fumigation time. VHP fumigation can be applied for in-situ biodecontamination of the HEPA filter unit as an alternative method to formaldehyde fumigation.

Curcumin Recovers Intracellular Lipid Droplet Formation Through Increasing Perilipin 5 Gene Expression in Activated Hepatic Stellate Cells In Vitro

The activation of hepatic stellate cells(HSCs)is a major event during hepatic fibrogenesis.Restoration of intracellular lipid droplet(LD)formation turns the activated HSC back to a quiescent state.Our previous studies have shown that curcumin suppresses HSC activation through increasing peroxisome proliferator-activated receptor,gamma(PPARγ)and 5′adenosine monophosphate-activated protein kinase(AMPK)activities.This study aims at evaluating the effect of curcumin on lipid accumulation in HSCs and hepatocytes,and further elucidating the underlying mechanisms.Now we showed that curcumin increased LD formation in activated HSCs and stimulated the expression of sterol regulatory element-binding protein and fatty acid synthase,and reduced the expression of adipose triglyceride lipase.Exogenous perilin5 expression in primary HSCs promoted LD formation.Perilipin 5 siRNA eliminated curcumin-induced LD formation in HSCs.These results suggest that curcumin recovers LD formation and lipid accumulation in activated HSCs by increasing perilipin 5 gene expression.Furthermore,inhibition of AMPK or PPARy activity blocked curcumin's effect on Plin5 gene expression and LD formation.Our results provide a novel evidence in vitro for curcumin as a safe,effective candidate to treat liver fibrosis.

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.

Feedforward loop profile among transcription factor, miRNA and mRNA in influenza A virus-infected mouse lungs

Purpose:In the present study,we focused on the 46 microRNAs and 719 genes in the microRNA-gene network,reported by us,and aimed to build a research blueprint of feedforward loops and reveal the key TFs in H1N1-infected mouse lung.Method:Based on microRNAs and genes in the microRNA-gene network previously reported by us,we used Jemboss software to find relationships between TFs and microRNAs(or genes),and then built a TF-microRNA-gene network exploiting the interactions between TFs and microRNAs(or genes).Next,we searched the sequences of above genes or microRNAs near the transcription start site(TSS)area,and then used the MatchTM algorithm to predict relevant TFs,and built the TF-Gene-Network.Result:We built a TF-microRNAgene network and exploreed eight key TFs,namely NF-AT1,GKLF,SRY,SOX10,AML1,MZF1,CRX and myogenin,in the network,and then constructed subgraphs of these eight TFs.Simultaneously,we predicted the possible target genes of microRNAs and identified the feedforward regulation relationship of possible TFs,microRNAs and mRNAs.The results showed that all eight factors with a score greater than 100 were TFs,namely NF-AT1,GKLF,SRY,SOX10,AML1,CRX,myogenin and MZF1.We then constructed subtables of the above eight TFs.Conclusion:In this study,TFs including NF-AT1,GKLF,SRY,SOX10,AML1,MZF1,CRX and myogenin showed the highest score(>100)not only in the TF-microRNA-gene network but also in feedforward loops,indicating that these eight TFs play the most important roles in mouse H1N1 influenza virus infection biology.

Effectiveness of chemical inactivation of infectious liquid biological waste:A randomized sample study of research laboratories in Switzerland

Laboratory wastewater has been suggested as an important escape route for microorganisms from research environments.Likely reasons for the unintentional release of laboratory organisms are shortcomings in the handling of infectious liquid biological waste(LBW)and inadequate inactivation procedures.We developed an analytical approach to investigate the use of chemical inactivation(CI)procedures in Swiss research laboratories by on-site random sampling of presumably inactivated infectious LBW and testing it for the presence of infectious lentiviruses(HIV-1)and adenoviruses(AdV).In addition,standard operating procedures(SOPs)for CI were collected and evaluated,and laboratorystaff knowledge of CI processes was assessed using a questionnaire.Although we found several deficiencies in the technical knowledge and training of laboratory staff on the CI of LBW,as documented by 27 returned questionnaires,no infectious viruses were detected in the eight LBW samples collected.Whilst we acknowledge that the number of LBW samples and SOPs is small,we conclude that CI of LBW containing infectious lentiviruses and adenoviruses does not result in the systematic release of considerable amounts of infectious viruses into the environment from research laboratories in Switzerland.

Assessing risk of Bombali virus spillover to humans by mutagenesis analysis of viral glycoprotein

Ebolaviruses,such as the highly pathogenic Ebola virus(EBOV),belong to the family Filoviridae and have caused out-breaks of severe and fatal hemorrhagic fever over the past years.Bombali virus(BOMV)is a newly discovered ebolavirus from bats with unknown potential to infect humans.For most ebolaviruses,the interaction between viral glycoprotein(GP)and host receptor Niemann-Pick C1(NPC1)is crucial for viral entry and determines host tropism.Here,we analyzed the BOMV GP-mediated virus entry into human cells using our recently developed EBOV transcription-and replication-competent virus-like particle(trVLP)system.We demonstrated that while BOMV GP can be efficiently incorporated into trVLPs,it is inefficient in mediating trVLP entry into human cells.However,BOMV GP-mediated virus entry into human cells can be significantly enhanced by a few mutations in the NPC1-binding domain of GP.Furthermore,we showed that these mutations increase the binding of BOMV-GP to human NPC1.In summary,our results suggested that although wild-type BOMV does not efficiently infect human cells,the emergence of mutations in viral GP may boost its ability to spill over to humans,highlighting the importance of monitoring BOMV GP evolution in preventing potential BOMV spillover events.

Current status and future challenges of high-level biosafety laboratories in China

High-level biosafety laboratories are safe and secure platforms which integrate reliable containment,well-trained personnel,and specific biosafety manuals and practices to protect researchers from being infected while manipulating microbial pathogens and prevent pathogens from being released into the outside environment.During the past decades,laboratories with different protection levels have been constructed and operated,the legal framework and a laboratory biosafety management system have been established,and these operational laboratories have played an essential role in combatting emerging and re-emerging infectious diseases in China.Here,we have summarized the significant achievements of high-level laboratory planning,construction,and operation in China,as well as the challenges that we face in the future.Additionally,we have briefly discussed the national plan for construction of highlevel biosafety laboratories from 2016-2025 and‘‘biosafety-motivated”management system.This review might be informative for further understanding the present situation and future function of high-level biosafety laboratories in China.

Quality management in a high-containment laboratory

The National Biosafety Laboratory level 4 in Wuhan was accredited according to the Guo Biao(National standard)19489:2008 standard in 2017 by the China National Accreditation Service for Conformity Assessment and was then licensed to store and handle microorganisms of risk group 4.The current aim of laboratory management is to accredit this high-containment laboratory according to the ISO/IEC 17025:2017 standard to set up a quality management system.To complement this standard,the ComitéEuropéen de Normalisation(CEN,i.e.,European Committee for Standardization)Working Agreement 15793:2011 is also used for safety and security against biorisks.The Chinese Guo Biao 19489:2008 standard is also taken into account to achieve two aims:to comply with Chinese legislation and to improve the level of detail in the laboratory’s documentation.Furthermore,the management proposes to apply Good Research Laboratory Practices(published by the China National Accreditation Service for Conformity Assessment),in which ethical rules are laid down.Only a well-integrated and well-managed quality management system can master the implementation of all these different guidelines.Nonetheless,it would be better if all the requirements were combined into one standard.Therefore,there is a real need for drafting a new standard or guideline specifically for containment laboratories.

Ebola virus VP35 perturbs type I interferon signaling to facilitate viral replication

As one of the deadliest viruses,Ebola virus(EBOV)causes lethal hemorrhagic fevers in humans and nonhuman primates.The suppression of innate immunity leads to robust systemic virus replication of EBOV,leading to enhanced transmission.However,the mechanism of EBOV-host interaction is not fully understood.Here,we identified multiple dysregulated genes in early stage of EBOV infection through transcriptomic analysis,which are highly clustered to Jak-STAT signaling.EBOV VP35 and VP30 were found to inhibit type I interferon(IFN)signaling.Moreover,exogenous expression of VP35 blocks the phosphorylation of endogenous STAT1,and suppresses nuclear translocation of STAT1.Using serial truncated mutations of VP35,N-terminal 1–220amino acid residues of VP35 were identified to be essential for blocking on type I IFN signaling.Remarkably,VP35 of EBOV suppresses type I IFN signaling more efficiently than those of Bundibugyo virus(BDBV)and Marburg virus(MARV),resulting in stable replication to facilitate the pathogenesis.Altogether,this study enriches understanding on EBOV evasion of innate immune response,and provides insights into the interplay between filoviruses and host.

Broadly neutralizing antibodies derived from the earliest COVID-19 convalescents protect mice from SARS-CoV-2 variants challenge

Coronavirus disease 2019(COviD-19)was first reported three years ago,when a group of individuals were infected with the original SARS-CoV-2 strain,based on which vaccines were developed.Here,we develop six human monoclonal antibodies(mAbs)from two elite convalescents in Wuhan and show that these mAbs recognize diverse epitopes on the receptor binding domain(RBD)and can inhibit the infection of SARS-CoV-2 original strain and variants of concern(VOCs)to varying degrees,including Omicron strains XBB and XBB.

Rapid Acquisition of High-Quality SARS-CoV-2 Genome via Amplicon-Oxford Nanopore Sequencing

Genome sequencing has shown strong capabilities in the initial stages of the COVID-19 pandemic such as pathogen identification and virus preliminary tracing.While the rapid acquisition of SARS-Co V-2 genome from clinical specimens is limited by their low nucleic acid load and the complexity of the nucleic acid background.To address this issue,we modified and evaluated an approach by utilizing SARS-Co V-2-specific amplicon amplification and Oxford Nanopore Prometh ION platform.This workflow started with the throat swab of the COVID-19 patient,combined reverse transcript PCR,and multi-amplification in one-step to shorten the experiment time,then can quickly and steadily obtain high-quality SARS-Co V-2 genome within 24 h.A comprehensive evaluation of the method was conducted in 42 samples:the sequencing quality of the method was correlated well with the viral load of the samples;high-quality SARS-Co V-2 genome could be obtained stably in the samples with Ct value up to 39.14;data yielding for different Ct values were assessed and the recommended sequencing time was 8 h for samples with Ct value of less than 20;variation analysis indicated that the method can detect the existing and emerging genomic mutations as well;Illumina sequencing verified that ultra-deep sequencing can greatly improve the single read error rate of Nanopore sequencing,making it as low as 0.4/10,000 bp.In summary,high-quality SARS-Co V-2 genome can be acquired by utilizing the amplicon amplification and it is an effective method in accelerating the acquisition of genetic resources and tracking the genome diversity of SARSCo V-2.

High-containment facilities and the role they play in global health security

Biological threats,whether naturally occurring or accidentally or deliberately released,have the potential to endanger lives and disrupt economies worldwide.Thus,high-containment protective measures should be implemented when handling bio-agents that can cause serious,highly contagious diseases,otherwise devastating pandemics can occur.In the fields of scientific investigation,healthcare,and product development,high-containment facilities play a critical role in preventing,detecting,and responding promptly and effectively to threats to global health security.In this paper,we present a summary of the main types of high-containment facilities,as well as their applications,challenges,and suggestions for the future.

SARS-CoV-2 infection induces testicular injury in Rhesus macaque

Dear Editor,COVID-19 is caused by the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),which is the seventh coronavirus known to infect humans.The COVID-19 pandemic has spread to more than 200countries,with more than 600 million confirmed cases and over 6.5million deaths as of September 2022,posing a huge challenge to global public health.

Correction to: Rapid Acquisition of High-Quality SARS-CoV-2 Genome via Amplicon-Oxford Nanopore Sequencing

Correction to:Virologica Sinica(2021)36:901-912 https://doi.Org/10.1007/sl 2250-021-00378-8 The original version of article contains errors in Figure 4,Supplementary Figure S2,Supplementary Table S2,and the corresponding description in part 3 of“Results”section.

Safety and security in the age of synthetic biology

Synthetic biology offers great potential for benefit to society,human health and agriculture;however,it also raises new concerns about safety and security.Application of established tools of biosafety and biosecurity along with strong individual and institutional leadership will help alleviate risks,but these may require refinement to address successfully the potential challenges of synthetic biology.Institutional biosafety committees(IBC)are key to providing institutional oversight and as technology evolves,especially in the case of synthetic biology where many different specialized fields may play a role.Membership of the IBC needs to adapt to ensure that sufficient knowledge and experience is available to evaluate projects and recognize potentially dangerous experiments.Recommendations regarding avian influenza gain of function studies may offer a valuable framework of points to consider relevant future studies involving synthetic biology.