Potent T cell Responses Induced by Single DNA Vaccine Boosted with Recombinant Vaccinia Vaccine

Plasmid DNA, an effective vaccine vector, can induce both cellular and humoral immune responses. However, plasmid DNA raises issues concerning potential genomic integration after injection. This issue should be considered in preclinical studies. Tiantan vaccinia virus (TV) has been most widely utilized in eradicating smallpox in China. This virus has also been considered as a successful vaccine vector against a few infectious diseases. Potent T cell responses through T-cell receptor (TCR) could be induced by three injections of the DNA prime vaccine followed by a single injection of recombinant vaccinia vaccine. To develop a safer immunization strategy, a single DNA prime followed by a single recombinant Tiantan vaccinia (rTV) AIDS vaccine was used to immunize mice. Our data demonstrated that one DNA prime/rTV boost regimen induced mature TCR activation with high functional avidity, preferential T cell Vβ receptor usage and high sensitivity to anti-CD3 antibody stimulation. No differences in T cell responses were observed among one, two or three DNA prime/rTV boost regimens. This study shows that one DNA prime/rTV boost regimen is sufficient to induce potent T cell responses against HIV.

DNA vaccine prime and replicating vaccinia vaccine boost induce robust humoral and cellular immune responses against MERS-CoV in mice

As of December 2022,2603 laboratory-identified Middle East respiratory syndrome coronavirus(MERS-CoV)infections and 935 associated deaths,with a mortality rate of 36%,had been reported to the World Health Organization(WHO).However,there are still no vaccines for MERS-CoV,which makes the prevention and control of MERS-CoV difficult.In this study,we generated two DNA vaccine candidates by integrating MERS-CoV Spike(S)gene into a replicating Vaccinia Tian Tan(VTT)vector.Compared to homologous immunization with either vaccine,mice immunized with DNA vaccine prime and VTT vaccine boost exhibited much stronger and durable humoral and cellular immune responses.The immunized mice produced robust binding antibodies and broad neutralizing antibodies against the EMC2012,England1 and KNIH strains of MERS-CoV.Prime-Boost immunization also induced strong MERS-S specific T cells responses,with high memory and poly-functional(CD107a-IFN-γ-TNF-α)effector CD8t T cells.In conclusion,the research demonstrated that DNA-Prime/VTT-Boost strategy could elicit robust and balanced humoral and cellular immune responses against MERS-CoV-S.This study not only provides a promising set of MERS-CoV vaccine candidates,but also proposes a heterologous sequential immunization strategy worthy of further development.

From a Deadly Disease to a Manageable Chronic Disease,HIV/AIDS Remains a Challenge for Mankind

Acquired immune deficiency syndrome(AIDS)is caused by human immunodeficiency virus(HIV),which was found by French scientists 40 years ago,12 years after AIDS was firstly reported by US scientists.2 TheHIV/AIDS pandemic has spread to every country of theworldwith over 84 million infections and more than 40 million deaths since the start of the epidemic.3 According to Joint United Nations Programme on HIV/AIDS(UNAIDS),there are 38.4 million people living with HIV/AIDS(PLWHA)in 2021.4 In order to realize the Ending AIDS Plan by 2030,the UNAIDS made the intermediate target of less than 500,000 new HIV infections and 500,000 AIDS-related deaths in 2020.5 In the 2021 United Nations(UN)75th General Assembly,the General Secretary reported that the 2020 target of Ending AIDS Plan was not reached,6 with 1.7 million HIV infections and 690,000 AIDS-related deaths in 2019.7 In order to bring the Ending AIDS Plan back on track,UNAIDS setup a higher goal of“95-95-95,”meaning the percentage for all PLWHA who know their HIV status.

Narrow bandgap molecular dyads incorporating Y-series acceptor backbones for efficient single-molecular organic solar cells

The performance of organic solar cells(OSCs)is mainly related to the bulk heterojunction(BHJ)microstructure of specific active layer systems,which is often in a metastable state.A promising strategy to address the abovementioned shortcomings of BHJs is to develop single-component active layer materials.Owing to the single-component small molecule materials with defined chemical structures generally exhibit poor absorption spectra,herein we first introduced narrow bandgap Y-series acceptors into the molecular skeleton of single-component materials,and designed two molecular dyads,SM-Et-1Y and SM-Et-2Y.The optical bandgaps(E_g~(opt)s)of the two dyads are 1.364 and 1.361 eV,respectively,which are much smaller than those of previously reported single-component molecules.Consequently,the SM-Et-2Y-based single-component OSCs(SCOSCs)showed a power conversion efficiency(PCE)of 5.07%,superior to SM-Et-1Y(2.53%),which is one of the highest PCEs reported for SCOSCs to date.Moreover,both SM-Et-1Y-and SM-Et-2Y-based devices exhibited excellent photo-stability,retaining over 90%of their initial performance after 250 h of continuous illumination.Our results provide a deeper understanding of the molecular backbone and a guiding principle for the rational design or selection of non-fullerene single-component materials with suitable donor/acceptor ratios.

Low-cost organic photovoltaic materials with great application potentials enabled by developing isomerized non-fused ring acceptors

Benefitting from low cost and simple synthesis,simple structured non-fused ring acceptors(NFRAs)and polymer donors are crucial for the application of organic solar cells(OSCs).Herein,two isomerized NFRAs,namely 4T-FCl FCl and 4T-2F2Cl,are designed with end-group engineering,which modulates the electrostatic potential distributions and crystallinity of acceptors,and accordingly,the A/A and D/A intermolecular interactions.The OSC based on 4T-2F2Cl with strong D/A interactions shows a record-high efficiency of 16.31%in blending with a low-cost polymer donor PTQ10,which shapes obviously improved bulkheterojunction(BHJ)networks blade-coated by non-halogenated solvent o-xylene,and thus significantly diminishes nonradiative recombination loss.A higher industrial figure of merit(i-FOM)of 0.46 for PTQ10:4T-2F2Cl in comparison with PTQ10:4T-FCl FCl(i-FOM=0.29)is demonstrated owing to its superior device efficiency and operational stability.Note that the i-FOM of PTQ10:4T-2F2Cl is the highest value for OSCs reported so far.This work deepens the synergistic effect of the A/A and D/A interactions on achieving desired bulk heterojunction morphology and demonstrates a printable photovoltaic system for low-cost,high-efficiency,stable,and eco-friendly OSCs.

The YDA-MKK4/MKK5-MPK3/MPK6 Cascade Functions Downstream of the RGF1-RGI Ligand-Receptor Pair in Regulating Mitotic Activity in Root Apical Meristem

The mitotic activity of root apical meristem(RAM)is critical to primary root growth and development.Previous studies have identified the roles of ROOT GROWTH FACTOR 1(RGF1),a peptide ligand,and its receptors,RGF1 INSENSITIVEs(RGIs),a clade of five leucine-rich-repeat receptor-like kinases,in promoting cell division in the RAM,which determines the primary root length.However,the downstream signaling components remain elusive.In this study,we identify a complete mitogen-activated protein kinase(MAPK or MPK)cascade,composed of YDA,MKK4/MKK5,and MPK3/MPK6,that functions downstream of the RGF1-RGI ligand-receptor pair.Similar to the rgi1/2/3/4/5 quintuple mutant,loss-of-function mutants of MPK3 and MPK6,MKK4 and MKK5,or YDA show a short-root phenotype,which is associated with reduced mitotic activity and lower expression of PLETHORA 1(PLT1)/PLT2 in the RAM.Furthermore,MPK3/MPK6 activation in response to exogenous RGF1 treatment is impaired in the rgi1/2/3/4/5 quintuple,yda single,and mkk4 m kk5 double mutants.Epistatic analyses demonstrated that the expression of constitutively active MKK4,MKK5,or YDA driven by the RGI2 promoter can rescue the short-root phenotype of the rgi1/2/3/4/5 mutant.Taken together,these results suggest that the YDA-MKK4/MKK5-MPK3/MPK6 cascade functions downstream of the RGF1-RGI ligand-receptor pair and upstream of PLT1/PLT2 to modulate the stem cell population and primary root growth in Arabidopsis.

Depletion and dysfunction of Vy,2Vo2 T cells in HIV disease： mechanisms, impacts and therapeutic implications

Infection with human immunodeficiency virus （HIV） disrupts the balance among yT cell subsets, with increasing Vo1＋ cells and substantial depletion of circulating Vo2＋ cells. Depletion is an indirect effect of HIV in CD4-negative Vo2 cells, but is specific for phosphoantigen-responsive subpopulations identified by the Vy2-Jy1.2 （also called Vy9-JyP） T cell receptor rearrangement. The extent of cell loss and recovery is related closely to clinical status, with highest levels of functional V cells present in virus controllers （undetectable viremia in the absence of antiretroviral therapy）. We review the mechanisms and clinical consequences for V cell depletion in HIV disease. We address the question of whether HIV-mediated V cell depletion, despite being an indirect effect of infection, is an important part of the immune evasion strategy for this virus. The important roles for V cells, as effectors and immune regulators, identify key mechanisms affected by HIV and show the strong relationships between V62 cell loss and immunodeficiency disease. This field is moving toward immune therapies based on targeting V cells and we now have clear goals and expectations to Ruide interventional clinical trials.

Neutrophil-derived heparin binding protein triggers vascular leakage and synergizes with myeloperoxidase at the early stage of severe burns(With video)

Background:Burn shock caused by vascular leakage is one of the main causes of high mortality in severe burn injury.However,the pathophysiological mechanism of vascular leakage is still unclear.The purpose of this study was to explore the molecular mechanism of vascular leakage in the early stage of severe burn and provide a new target for the treatment of severe burns.Methods:Neutrophils were isolated from human peripheral blood by magnetic beads sorting.ELISA was used to detect neutrophil-derived granule proteins and glycocalyx injury products in plasma.The vascular leakage and neutrophil movement were assessed by in vivo laser confocal imaging in mice,and high-quality video were provided.Adhesion-related molecules were investigated by qRT-PCR.The damage to glycocalyx of mice vascular endothelial cellswas observed by transmission electron microscope and scanning electron microscope.Proteomic analysis,flow cytometry and immunofluorescence were used to further study the relationship between human peripheral blood neutrophil-derived hypochlorite(HOCl)and CD44 of human vascular endothelial cells.Results:In this study,we found that rapidly increasing activated neutrophils secrete heparin binding protein(HBP)andmyeloperoxidase(MPO)after severe burn injury.Increased HBP triggers vascular leakage with synergy of MPO,results in systemic edema and burn shock.Furthermore,we found that the MPO catalytic product HOCl but not MPO triggers CD44 extracellular domain shedding from vascular endothelial cells to damage the glycocalyx.Damage to the glycocalyx results in firm adhesion of neutrophils and increases vascular leakage.However,MPO inhibitors partially protect the glycocalyx of vascular endothelial cells.The combination of HBP and MPO inhibitors markedly reduces vascular leakage and systemic edema in the early stage of severe burns.Conclusions:Taken together,these data reveal that neutrophil-derived HBP and MPO play an important synergies role in triggering vascular leakage at the early stage of severe burns.Targeted intervention in these two biomolecules may introduce new strategies for helping to reduce large amount of fluid loss and subsequent burn shock.

Higher levels of circulating monocyte-platelet aggregates are correlated with viremia and increased sCD 163 levels in HIV- 1 infection

Increased levels of monocyte-platelet aggregates （MPAs） are reported to be highly correlated with cardiovascular events. In this study, the MPA levels in different monocyte subsets and the associations between MPA levels, HIV-1 viremia and monocyte activation were evaluated during HIV-1 infection. The results showed that the percentages of MPAs in all three monocyte subsets were higher in HIV-l-infected subjects than in healthy controls, and were associated with the plasma viral load in the non-classical and intermediate monocyte subsets. The plasma levels of sCD14 and sCD163 were upregulated in HIV-1 infection and were positively associated with viral loads and negatively associated with CD4 counts. P-selectin glycoprotein ligand-1 （PSGL-1） was shown to be expressed at significantly lower levels on all three monocyte subsets and was negatively correlated with the sCD163 level. The MPA level was correlated with the levels of plasma sCD163 but negatively correlated with CD163 and PSGL-1 on all three monocyte subsets. An elevated immune activation status was correlated with increased MPA formation, underlying the potential interaction between monocyte activation and MPA formation. This interaction may be related to a higher thromboembolic risk in patients infected with HIV-I.

Changing Proportions of HIV-1 Subtypes and Transmitted Drug Resistance Among Newly Diagnosed HIV/AIDS Individuals—China,2015 and 2018

Introduction:With the expansion of human immunodeficiency virus(HIV)antiretroviral therapy(ART),HIV drug resistance is becoming more and more serious.This study describes the changing prevalence of HIV-1 subtypes and transmitted drug resistance(TDR)among newly diagnosed individuals in China,2015 and 2018.Methods:A total of 8,980 individuals in 2015 and 2018 from 31 provincial-level administrative divisions(PLADs)were enrolled in this study.Viral RNAs were amplified and sequenced using an in-house polymerase chain reaction(PCR)protocol.The Stanford HIV Drug Resistance Database(HIVdb)was used to predict susceptibility to 12 antiretroviral drugs.Results:The prevalence of TDR was not significantly increased over time.The prevalence of TDR was 3.8%and 4.4%in 2015 and 2018,respectively(P=0.13).The prevalence of CRF55_01B increased from 2.3%in 2015 to 3.9%in 2018(P<0.001).The drug resistance prevalence of nonnucleoside reverse transcriptase inhibitors(NNRTI)increased from 2.4%in 2015 to 3.3%in 2018(P<0.01).The prevalence of E138(P<0.001),H221(P=0.03),and V179(P<0.001)mutations increased from 0.30%,0.09%,and 0.70%in 2015 to 1.10%,0.30%,and 1.70%in 2018,respectively.Conclusions:HIV drug resistance affects the effect of antiretroviral treatment,so the monitoring of HIV TDR should be strengthened to control the transmission of HIV drug resistance.

Neutrophil extracellular traps contribute to myofibroblast differentiation and scar hyperplasia through the Toll-like receptor 9/nuclear factor Kappa-B/interleukin-6 pathway

Background:Inflammation is an important factor in pathological scarring.The role of neutrophils,one of the most important inflammatory cells,in scar hyperplasia remains unclear.The purpose of this article is to study the correlation between neutrophil extracellular traps(NETs)and scar hyperplasia and identify a new target for inhibiting scar hyperplasia.Methods:Neutrophils were isolated from human peripheral blood by magnetic-bead sorting.NETs in plasma and scars were detected by enzyme-linked immunosorbent assays(ELISAs),immunofluorescence and flow cytometry.Immunohistochemistry was used to assess neutrophil(CD66B)infiltration in hypertrophic scars.To observe the entry of NETs into fibroblasts we used immunofluorescence and flow cytometry.Results:We found that peripheral blood neutrophils in patients with hypertrophic scars were more likely to form NETs(p<0.05).Hypertrophic scars showed greater infiltration with neutrophils and NETs(p<0.05).NETs activate fibroblasts in vitro to promote their differentiation and migration.Inhibition of NETs with cytochalasin in wounds reduced the hyperplasia of scars in mice.We induced neutrophils to generate NETs with different stimuli in vitro and detected the proteins carried by NETs.We did not find an increase in the expression of common scarring factors[interleukin(IL)-17 and transforming growth factor-β(TGF-β),p>0.05].However,inhibiting the production of NETs or degrading DNA reduced the differentiation of fibroblasts intomyofibroblasts.In vitro,NETs were found to be mediated by Toll-like receptor 9(TLR-9)in fibroblasts and further phosphorylated nuclear factor Kappa-B(NF-κB).We found that IL-6,which is downstream of NF-κB,was increased in fibroblasts.Additionally,IL-6 uses autocrine and paracrine signaling to promote differentiation and secretion.Conclusions:Our experiments found that NETs activate fibroblasts through the TLR-9/NF-κB/IL-6 pathway,thereby providing a new target for regulating hypertrophic scars.

Propagation analysis and prediction of the COVID-19

Based on the official data modeling,this paper studies the transmission process of the Corona Virus Disease 2019(COVID-19).The error between the model and the official data curve is quite small.At the same time,it realized forward prediction and backward inference of the epidemic situation,and the relevant analysis help relevant countries to make decisions.

Treatment for HIV prevention study in southwestern areas of China

Background:China has ambitious to achieve significant reductions in HIV transmission and HIV-related mortality by adopting the World Health Organization's“Treat All”approach.Such a prevention strategy is needed future study on regional scale.Methods:An observational cohort study of HIV epidemiology and treatment databases was used to study the effectiveness of antiretroviral therapy on the transmission of HIV in serodiscordant couples in Guangxi of China.Results:A total of 7713 couples were entered into the cohort study analysis which included 1885 couples in the treatment-naive cohort and 5828 couples in the treated cohort.During the follow-up of 18985.29 person-years from 2003 to 2014,the average incidence of HIV was 2.4 per 100 person-years(95%CI 2.1e2.6).HIV seroincidence rate was significantly higher among the treatment naive group(4.2 per 100 person-years,3.7e4.8)compared with the on treatment group(1.6 per 100 person-years,1.3e1.8).An overall 45%reduction in risk of HIV transmission among serodiscordant couple was associated with ART treatment(adjusted Hazard Ratio[HR]0.55,95%Confidence Interval[CI]0.44e0.69).Treatment prevention had significantly effectiveness for most baseline characteristics of index partners,such as for male,female,age above 25 years,education below high school,farmer,infected by heterosexual intercourse.Conclusion:Treatment-as-prevention can be implemented in the real-world on a national or regional scale,but ART adherence and comprehensive harm reduction while implementing this strategy require further study.

Relationship Between Drug Resistance and Death in HIV-Infected Patients Receiving Antiretroviral Therapy—7 PLADs,China,2010−2019

Summary What is already known about this topic?With increasing coverage of antiretroviral therapy(ART)for HIV-infected patients,more and more attention has been paid to the impact of HIV drug resistance on death in those patients in China.What is added by this report?Among HIV-infected patients receiving ART,the risk of death is higher in patients with HIV drug resistance[adjusted odds ratio(AOR)=4.25.

A mathematical modeling study of the HIV epidemics at two rural townships in the Liangshan Prefecture of the Sichuan Province of China

Background:As a response to a severe HIV epidemic in the Liangshan Prefecture,one of the worst in China,population based HIV interventions,including two population-wide HIV screening,have been carried out since 2005 at two townships in a remote mountainous region of Liangshan.The objective of our mathematical modeling study is to assess the temporal dynamics of the HIV epidemic in the two townships based on the data collected in the study area during the period 2005e2010.Methods:A mathematical model was set up to describe the population dynamics of HIV transmission in study area.The model was calibrated by fitting it to the HIV testing and treatment data from 2005 to 2008.Validation of the model was done by comparing its predicted value of HIV prevalence in 2010 to the prevalence data obtained in the 2010 population wide HIV testing.The validated model was used to produce estimation of HIV incidence,prevalence and death.Results:Our model estimations show that population-based HIV interventions have significantly slowed down the rise of the HIV epidemic in the two townships.Over the five-year period from 2005 to 2010,the year-over-year rate of increase in HIV incidence,prevalence,and death has declined by 91.5%,28.7%,and 52.3%,respectively.Conclusion:Mathematical models,when integrated with epidemiological and surveillance data,can be an effective tool for predicting the temporal dynamics of HIV and assessing the impacts of HIV interventions.

Premature Stop Codon at Residue 101 within HIV-1 Rev Does Not Influence Viral Replication of Clade BC but Severely Reduces Viral Fitness of Clade B

HIV-1 Rev is an accessory protein that plays a key role in nuclear exportation,stabilization,and translation of the viral mRNAs.Rev of HIV-1 clade BC often shows a truncation of 16 AAs due to a premature stop codon at residue 101.This stop codon presents the highest frequency in clade BC and the lowest frequency in clade B.In order to discover the potential biological effect of this truncation on Rev activity and virus replication of clade BC,we constructed Rev expression vectors of clade BC with or without 16 AAs within C-terminal separately,and replaced the stop codon by Q in a CRF07_BC infectious clone.We found that 16 AAs truncation had no effect on expression and activity of Rev in clade BC.Also,the mutation from the stop codon to Q had no effect on virus replication of clade BC.Next,to investigate the effect of this truncation on Rev activity and replication capacity of clade B,Rev expression vectors of clade B carrying or lacking 16 AAs in C-terminal were constructed respectively,and residue Q at position 101 within Rev was substituted by the stop codon in a clade B infectious clone.It was found that 16 AAs truncation significantly down-regulated Rev expression and impaired clade B Rev activity.Furthermore,a Q-to-stop codon substitution within Rev significantly reduced viral replication fitness of clade B.These results indicate that the premature stop codon at residue 101 within Rev exerts diverse impact on viral replication among different HIV-1 clades.

SARS-CoV-2 vaccine research and immunization strategies for improved control of the COVID-19 pandemic

The record speed at which Chinese scientists identified severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)and shared its genomic sequence with the world,has greatly facilitated the development of coronavirus disease(COVID-19)diagnostics,drugs,and vaccines.It is unprecedented in pandemic control history to develop a dozen successful vaccines in the first year and to immunize over half of the global population in the second year,due to the efforts of the scientific community,biopharmaceutical industry,and regulatory agencies worldwide.The challenges are both great and multidimensional due to the rapid emergence of virus variants and waning of vaccine immunity.Vaccination strategies need to adapt to these challenges to keep population immunity above the herd immunity threshold,by increasing vaccine coverage,especially for older adults and young people,and providing timely booster doses with homologous or heterologous vaccine boosts.Further research should be undertaken to develop more effective vaccines against SARS-CoV-2 variants and to understand the best prime-boost vaccine combinations and immunization strategies to provide sufficient and sustainable immune protection against COVID-19.

Internet search data could Be used as novel indicator for assessing COVID-19 epidemic

The pandemic of the coronavirus disease(COVID-19)poses a huge challenge all countries,since no one is well prepared for it.To be better prepared for future pandemics,we evaluated association between the internet search data with reported COVID-19 cases to verify whether it could become an early indicator for emerging epidemic.After the keyword filtering and Index composition,we found that there were close correlations between Composite Index and suspected cases for COVID-19(r?0.921,P<0.05).The Search Index was applied for the Autoregressive Integrated Moving Average with Exogenous Variables(ARIMAX)model to quantify the relationship.Compared with the model based on surveillance data only,the ARIMAX model had smaller Akaike Information Criterion(AIC?403.51)and the most accurate predictive values.Overall,the Internet search data could serve as a convenient indicator for predicting the epidemic and to monitor its trends.

Global Transmission of Human Immunodeficiency Virus 1 Subtype C and Its Impact on the Circulation of B/C Recombination Strains in China

This study aimed to reconstruct the origin and worldwide epidemic history of human immunodeficiency virus(HIV)1 subtype C and comprehend how HIV-1 subtype C was introduced into and spread throughout China in the formof B/C recombinant strains.Envelope sequences ofHIV-1 subtype C and some other subtypes deposited before December 31,2020 were downloaded from the Los Alamos HIV Database and the Chinese National Center for AIDS/STD Control and Prevention Database.The available sequences were screened for quality,and Bayesian analysis was used to build the maximum clade credibility evolutionary tree to analyze and judge the origin and spread of HIV-1 subtype C.HIV-1 subtype C originated in central Africa around 1952,then spread to southern Africa around 1969 and to eastern Africa around 1973.HIV-1 subtype C fromsouthern Africa was introduced into India in 1977.HIV-1 subtype C of eastern Africa was introduced into Brazil in 1987.Indian HIV-1 subtype C was exported to China in three migration events during the period from 1986 to 1989.The two predominant recombinants in China(CRF07_BC and CRF08_BC)emerged in 1988 and 1990,respectively.Other B/C recombinants,namely,CRF64_BC,CRF61_BC and CRF62_BC,originated in 1993,2002 and 2000,respectively.Our study has reconstructed the global origin and evolutionary history of HIV-1 subtype C.In addition,our study demonstrated that the Chinese HIV-1 subtype C originated from three related Indian lineages around the mid to late 1980s and,since then,has formed some B/C recombinants with subtype B that caused a widespread epidemic in China.

Spray characteristics and controlling mechanism of fuel containing CO2

This paper presents studies of spray characteristics and controlling mechanism of fuel containing CO2. Using diesel fuel containing CO2 gas, experiments were conducted on diesel hole-type nozzles and simple nozzles. The steady spray and transient spray characteristics were observed and measured by instantaneous shadowgraphy, high-speed photography, phase Doppler anemometry （PDA） and LDSA respectively. The effects of CO2 concentration in the fuel, the injection pressure, the nozzle L/D ratio, surrounding gas pressure and temperature on the atomization behavior and spray pattern were evaluated. The results show that the injection of fuel containing CO2 can greatly improve the atomization and produce a parabolic-shaped spray; and the CO2 gas concentration, surrounding gas pressure, temperature and nozzle config- uration have dominant influences on spray characteristics of the fuel containing CO2. New insight into the controlling mechanism of atomization of the fuel containing CO2 was provided.