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Activating long-lived room temperature phosphorescence (RTP) in the aqueous environment and thus realizing matrix-free, anti-oxygen, and time-resolved information encryption and cellular imaging remain a great challenge. Here, we fabricated three types of carbon dots (C-dots), i.e., fluorescent C-dots (F-C-dots) and two types of phosphorescent C-dots denoted as Pw-C-dots and Py-C-dots by a one-pot strategy. Their formation was attributed to the difference in the decarboxylation degree at high temperatures using trimesic acid (TMA) as a sole precursor. Unexpectedly, the yield reached as high as ∼92%, and the proportions were ∼27% for F-C-dots, ∼17% for Pw-C-dots, and ∼56% for Py-C-dots. These nanomaterials could help implement carbon peaking and carbon neutrality. Both green RTP of the two C-dots resulted from the small energy gap (Δ

The meningococcal serogroup B-factor H binding protein vaccine (MenB-FHbp) is licensed for use in children aged 10 years or older for protection against invasive serogroup B meningococcal disease. Because young children are at increased risk of invasive meningococcal disease, MenB-FHbp clinical data in this population are needed.We conducted two phase 2 randomised, controlled, observer-blinded studies including healthy toddlers (age 12-23 months) across 26 Australian, Czech, Finnish, and Polish centres, and older children (age 2-9 years) across 14 Finnish and Polish centres. Exclusion criteria included previous vaccinations against serogroup B meningococcus or hepatitis A virus (HAV), and chronic antibiotic use. Toddlers were randomly allocated (2:1) via an interactive response technology system to receive either 60 μg or 120 μg MenB-FHbp or HAV vaccine and saline (control). Older children were randomly allocated (3:1) to receive 120 μg MenB-FHbp or control, with stratification by age group (2-3 years and 4-9 years). All vaccinations were administered as three doses (0, 2, and 6 months, with only saline given at 2 months in the control group). Toddlers who received 120 μg MenB-FHbp could receive a 120 μg booster dose 24 months after the end of the primary series. The percentages of participants with serum bactericidal activity using human complement (hSBA) titres at or above the lower limit of quantification (LLOQ; all greater than the 1:4 correlate of protection) against four test strains of serogroup B meningococcus 1 month after the third dose (primary immunogenicity endpoint) were measured in the evaluable immunogenicity populations (participants who received the vaccine as randomised, had available and determinate hSBA results, and had no major protocol violations). Not all participants were tested against all strains because of serum sample volume constraints. The frequencies of reactogenicity and adverse events after each dose were recorded in the safety population (all participants who received at least one dose and had safety data available). These studies are registered with ClinicalTrials.gov (NCT02534935 and NCT02531698) and are completed.Between Aug 31, 2015, and Aug 22, 2016, for the toddler study and between Aug 27, 2015, and March 7, 2016, for the older children study, we enrolled and randomly allocated 396 toddlers (60 μg MenB-FHbp group n=44; 120 μg MenB-FHbp group n=220; control group n=132) and 400 older children (120 μg MenB-FHbp group n=294; control group n=106). 1 month after the third dose, the proportions of participants with hSBA titres at or above the LLOQ ranged across test strains from 85·0% (95% CI 62·1-96·8; 17 of 20 participants) to 100·0% (82·4-100·0; 19 of 19) in toddlers receiving 60 μg MenB-FHbp, and from 71·6% (61·4-80·4; 68 of 95) to 100·0% (96·2-100·0; 95 of 95) in toddlers receiving 120 μg MenB-FHbp, and from 79·1% (71·2-85·6; 106 of 134) to 100·0% (97·4-100·0; 139 of 139) in children aged 2-9 years receiving 120 μg MenB-FHbp. hSBA titres peaked at 1 month after the third primary dose of MenB-FHbp and then declined over time. 24 months after the third dose in the toddler study, the proportions with hSBA titres at or above the LLOQ ranged from 0·0% (0·0-17·6; 0 of 19 participants) to 41·2% (18·4-67·1; seven of 17) in those who received 60 μg MenB-FHbp and from 3·7% (0·8-10·4; three of 81) to 22·8% (14·1-33·6; 18 of 79) in those who received 120 μg MenB-FHbp. 1 month after the booster dose in toddlers, the proportions with hSBA titres at or above the LLOQ were higher than at 1 month after the primary series. MenB-FHbp reactogenicity was mostly transient and of mild to moderate severity. Adverse event frequency was similar between the MenB-FHbp and control groups and less frequent following MenB-FHbp booster than following primary doses. Two participants from the toddler study (both from the 120 μg MenB-FHbp group) and four from the older children study (three from the 120 μg MenB-FHbp group and one from the control group) were withdrawn from the study because of adverse events.MenB-FHbp was well tolerated and induced protective immune responses in a high proportion of participants. These findings support a favourable MenB-FHbp immunogenicity and reactogenicity profile in young children, a population at increased risk of adverse invasive meningococcal disease outcomes.Pfizer.

Au and PdOx show a synergistic effect on the CeO2 surface for the photo-assisted catalytic ozonation of toluene.

Anxiety is a normal and transitory emotional state that allows the organisms to cope well with the real or perceived threats, while excessive or prolonged anxiety is a key characteristic of anxiety disorders. We have recently revealed that prolonged anxiety induced by chronic stress is associated with the circuit-varying dysfunction of basolateral amygdala projection neurons (BLA PNs). However, it is not yet known whether similar mechanisms also emerge for acute stress-induced, short-lasting increase of anxiety. Here, using a mouse model of acute restraint stress (ARS), we found that ARS mice showed increased anxiety-like behavior at 2 h but not 24 h after stress, and this effect was accompanied by a transient increase of the activity of BLA PNs. Specifically, ex vivo patch-clamp recordings revealed that the increased BLA neuronal activity did not differ among the distinct BLA neuronal populations, regardless of their projection targets being the dorsomedial prefrontal cortex (dmPFC) or elsewhere. We further demonstrated that such effects were mainly mediated by the enhanced presynaptic glutamate release in dmPFC-to-BLA synapses but not lateral amygdala-to-BLA ones. Furthermore, while optogenetically weakening the presynaptic glutamate release in dmPFC-to-BLA synapses ameliorated ARS-induced anxiety-like behavior, strengthening the release increased in unstressed mice. Together, these findings suggest that acute stress causes short-lasting increase in anxiety-like behavior by facilitating synaptic transmission from the prefrontal cortex to the amygdala in a circuit-independent fashion.

Japanese encephalitis virus (JEV), an important neurotropic pathogen, belongs to the genus Flavivirus of the family Flaviviridae and has caused huge threat to public health. It is still obscure regarding the functions of stem-loop (SL) and dumbbell (DB) domains of JEV 3��� UTR in viral replication and virulence. In the current study, using the infectious clone of JEV SA14 strain as a backbone, we constructed a series of deletion mutants of 3��� UTR to investigate their effects on virus replication. The results showed that partial deletions within SL or DB domain had no apparent effects on virus replication in both mammalian (BHK-21) and mosquito (C6/36) cells, suggesting that they were not involved in viral host-specific replication. However, the entire SL domain deletion (��VR) significantly reduced virus replication in both cell lines, indicating the important role of the complete SL domain in virus replication. The revertant of ��VR mutant virus was obtained by serial passage in BHK-21 cells that acquired a duplication of DB domain (DB-dup) in the 3��� UTR, which greatly restored virus replication as well as the capability to produce the subgenomic flavivirus RNAs (sfRNAs). Interestingly, the DB-dup mutant virus was highly attenuated in C57BL/6 mice despite replicating similar to WT JEV. These findings demonstrate the significant roles of the duplicated structures in 3��� UTR in JEV replication and provide a novel strategy for the design of live-attenuated vaccines.

Floccules are another major form of microbial aggregates in aerobic granular sludge systems. Previous studies mainly attributed the persistence of floccules to their relatively faster nutrient uptake and higher growth rate over aerobic granules; however, they failed to unravel the underlying mechanism of the long-term coexistence of these two aggregates. In this work, the existence and function of the floccules in an aerobic granule-dominated sequencing batch reactor were investigated from the view of quorum sensing (QS) and quorum quenching (QQ). The results showed that though the floccules were closely associated with the granules in terms of similar community structures (including the QS- and QQ-related ones), they exhibited a relatively higher QQ-related activity but a lower QS-related activity. A compatible proportion of floccules might be helpful to maintain the QS-related activity and keep the granules stable. In addition, the structure difference was demonstrated to diversify the QS- and QQ-related activities of the floccules and the aerobic granules. These findings could broaden our understanding of the interactions between the coexistent floccules and granules in aerobic granule-dominated systems and would be instructive for the development of the aerobic granular sludge process.

Two three-dimensional (3D) nickel-based metal-organic frameworks (Ni(II)-MOFs), {[Ni(adc)(phen)(H2O)]}n (1, H2adc = 9,10-anthracenedicarboxylic acid, phen = 1,10-phenanthroline) and {[Ni(adc)(bpy)]}n (2, bpy = 4,4’-bipyridine), were synthesized via a mixed-ligand strategy under hydrothermal and solvothermal conditions. MOF 1 could be simplified as a binodal tfc topology, while 2 possesses a three-fold interpenetrated framework, which is composed of a 4-connected dia network. The phase purity, molecular vibrations, and thermal stability of 1 and 2 have been characterized by powder X-ray diffraction (PXRD), infrared spectroscopy (IR), and thermogravimetric analysis (TG). Further electrochemical experiments reveal that 1 and 2 exhibit good methanol/ethanol oxidation (MOR/EOR) activities, with the optimal current density reaching up to 21.2 (EOR: 25.3) and 15.8 (EOR: 15.3) mA cm−2, respectively.

Genetic engineering is promising to expand the application scope of exoelectrogens in energy and environmental applications and plasmid vectors, as one type of fundamental tool, are intensively applied. Antibiotics are widely utilized for plasmid selection and maintenance; however, their utilization suffers from environmental concerns on the spread of resistance genes, elevated costs, inevitable genotypic instability and phenotypic heterogeneity. In this work, we establish an auxotrophic complementation system for stable plasmid maintenance without antibiotic association, in Shewanella oneidensis, an attractive model exoelectrogen. A plasmid-transition strategy is designed to facilitate the rapid and efficient construction of the auxotrophic complementation system. Such a system not only enables the same intensive gene expression as the conventional antibiotic-associated plasmid system but also exhibits remarkably superior robustness and stability. With this system, the menaquinone pool of the extracellular respiratory chain is enhanced first independently and further synergized by engineering the Mtr conduit, leading to significantly promoted extracellular electron transfer (EET) outputs (up to 10.33- and 2.97-fold improvement in the maximum current density and the maximum output voltage) and heavy metal Cr(VI) reduction ability (5.15-fold improvement). This work provides a robust and stable platform to engineer exoelectrogens for environmental applications.

Recently LHCb experimental group find an exotic state $T^+_{cc}$ from the process $p\bar{p} \to D^0D^0\pi^+ + X$. A key question is if it is just a molecule or may have confined tetraquark ingredient. To investigate this, different methods are taken, including two channel ($D^{*+}D^0$ and $D^{*0}D^+$) K-matrix unitarization and single channel Flatt\'e-like parametrization method analysed by pole counting rule and spectral density function sum rule. It demonstrates that $T^+_{cc}$ is a molecular state, though the possibility that there may exist elementary ingredient can not be excluded, by rough analysis on its production rate.

The 4D Millimeter wave (mmWave) radar is a promising technology for vehicle sensing due to its cost-effectiveness and operability in adverse weather conditions. However, the adoption of this technology has been hindered by sparsity and noise issues in radar point cloud data. This paper introduces spatial multi-representation fusion (SMURF), a novel approach to 3D object detection using a single 4D imaging radar. SMURF leverages multiple representations of radar detection points, including pillarization and density features of a multi-dimensional Gaussian mixture distribution through kernel density estimation (KDE). KDE effectively mitigates measurement inaccuracy caused by limited angular resolution and multi-path propagation of radar signals. Additionally, KDE helps alleviate point cloud sparsity by capturing density features. Experimental evaluations on View-of-Delft (VoD) and TJ4DRadSet datasets demonstrate the effectiveness and generalization ability of SMURF, outperforming recently proposed 4D imaging radar-based single-representation models. Moreover, while using 4D imaging radar only, SMURF still achieves comparable performance to the state-of-the-art 4D imaging radar and camera fusion-based method, with an increase of 1.22% in the mean average precision on bird's-eye view of TJ4DRadSet dataset and 1.32% in the 3D mean average precision on the entire annotated area of VoD dataset. Our proposed method demonstrates impressive inference time and addresses the challenges of real-time detection, with the inference time no more than 0.05 seconds for most scans on both datasets. This research highlights the benefits of 4D mmWave radar and is a strong benchmark for subsequent works regarding 3D object detection with 4D imaging radar.

HLA-A*24:02:160 differs from HLA-A*24:02:01:01 by one nucleotide in exon 3. This article is protected by copyright. All rights reserved.

The nucleophilic superoxide radical (O

Background Chronic stress exposure increases the risk of mental health problems such as anxiety and depression. The medial prefrontal cortex (mPFC) is a hub for controlling stress responses through communicating with multiple limbic structures, including the basolateral amygdala (BLA) and nucleus accumbens (NAc). However, considering the complex topographical organization of the mPFC neurons in different subregions (dmPFC vs. vmPFC) and across multiple layers (Layer II/III vs. Layer V), the exact effects of chronic stress on these distinct mPFC output neurons remain largely unknown. Results We first characterized the topographical organization of mPFC neurons projecting to BLA and NAc. Then, by using a typical mouse model of chronic restraint stress (CRS), we investigated the effects of chronic stress on the synaptic activity and intrinsic properties of the two mPFC neuronal populations. Our results showed that there was limited collateralization of the BLA- and NAc-projecting pyramidal neurons, regardless of the subregion or layer they were situated in. CRS significantly reduced the inhibitory synaptic transmission onto the BLA-projecting neurons in dmPFC layer V without any effect on the excitatory synaptic transmission, thus leading to a shift of the excitation-inhibition (E-I) balance toward excitation. However, CRS did not affect the E-I balance in NAc-projecting neurons in any subregions or layers of mPFC. Moreover, CRS also preferentially increased the intrinsic excitability of the BLA-projecting neurons in dmPFC layer V. By contrast, it even caused a decreasing tendency in the excitability of NAc-projecting neurons in vmPFC layer II/III. Conclusion Our findings indicate that chronic stress exposure preferentially modulates the activity of the mPFC-BLA circuit in a subregion (dmPFC) and laminar (layer V) -dependent manner.

The analyses of the LHCb data on $X(6900)$ found in di-$J/\psi$ and $J/\psi \psi(3686)$ systems are performed using a momentum-dependent {Flatt\'{e}}-like parameterization. The use of the pole counting rule and spectral density function sum rule give consistent conclusions that $X(6900)$ may not be a molecule of $J/\psi \psi(3686)$. Nevertheless it is still possible that $X(6900)$ be a molecule of higher states, such as $J/\psi\psi(3770)$, $\chi_{c0}\chi_{c2}$, etc., Comment: 7pages with 9 figures

Clarifying the reaction pathways at the solid–water interface and in bulk water solution is of great significance for the design of heterogeneous catalysts for selective oxidation of organic pollutants. However, achieving this goal is daunting because of the intricate interfacial reactions at the catalyst surface. Herein, we unravel the origin of the organic oxidation reactions with metal oxide catalysts, revealing that the radical-based advanced oxidation processes (AOPs) prevail in bulk water but not on the solid catalyst surfaces. We show that such differing reaction pathways widely exist in various chemical oxidation (e.g., high-valent Mn 3+ and MnO X ) and Fenton and Fenton-like catalytic oxidation (e.g., Fe 2+ and FeOCl catalyzing H 2 O 2 , Co 2+ and Co 3 O 4 catalyzing persulfate) systems. Compared with the radical-based degradation and polymerization pathways of one-electron indirect AOP in homogeneous reactions, the heterogeneous catalysts provide unique surface properties to trigger surface-dependent coupling and polymerization pathways of a two-electron direct oxidative transfer process. These findings provide a fundamental understanding of catalytic organic oxidation processes at the solid–water interface, which could guide the design of heterogeneous nanocatalysts.

Largemouth bass (LMB) production exceeded 0.7 million tons in 2021 and has become one of the most important freshwater aquaculture species in China. The stable and fixed culture cycle led to regular and drastic price fluctuation during the past decade. Strong price fluctuation provides opportunities and challenges for the LMB industry, and out-of-season spawning (OSS) and culture will provide technical support for the opportunities. To induce OSS at a low cost, we established a controllable recirculating system that allows precise thermo-photoperiod manipulation. In the system, four experimental groups were assigned, 18NP (18°C overwintering water temperature, natural photoperiod), 18CP (18°C overwintering water temperature, controlled photoperiod), 16CP (16°C overwintering water temperature, controlled photoperiod), and NTNP (natural water temperature and natural photoperiod), to determine the effects of chilling temperature and photoperiod on spawning performance. OSS was observed in all the experimental groups without significant differences, except NTNP. The manipulated broodstock can re-spawn 3 months later in the next spring in advance. Further analysis of the volume percentage of different stages of oocytes provides a base for excellent regression between the volume percentage of the primary growth stage, cortical alveoli stage, vitellogenesis/maturation stage, and gonadal development/maturation. The results suggest that the volume percentage of oocytes is a better indicator of gonadal development and maturation than the gonadosomatic index. We also found that LMB prefers palm fiber as a spawning nest over gravel. The findings of this work provide important technique guidance for practical OSS of the LMB aquaculture industry and standardization of ovary development and maturation in fish with asynchronous developmental oocytes.

Background Coronavirus disease 2019 (COVID-19) can involve persistence, sequelae, and other clinical complications that last weeks to months to evolve into long COVID-19. Exploratory studies have suggested that interleukin-6 (IL-6) is related to COVID-19; however, the correlation between IL-6 and long COVID-19 is unknown. We designed a systematic review and meta-analysis to assess the relationship between IL-6 levels and long COVID-19. Methods Databases were systematically searched for articles with data on long COVID-19 and IL-6 levels published before September 2022. A total of 22 published studies were eligible for inclusion following the PRISMA guidelines. Analysis of data was undertaken by using Cochran's Q test and the Higgins I-squared (I2) statistic for heterogeneity. Random-effect meta-analyses were conducted to pool the IL-6 levels of long COVID-19 patients and to compare the differences in IL-6 levels among the long COVID-19, healthy, non-postacute sequelae of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection (non-PASC), and acute COVID-19 populations. The funnel plot and Egger's test were used to assess potential publication bias. Sensitivity analysis was used to test the stability of the results. Results An increase in IL-6 levels was observed after SARS-CoV-2 infection. The pooled estimate of IL-6 revealed a mean value of 20.92 pg/ml (95% CI = 9.30–32.54 pg/ml, I2 = 100%, P CI = 5.75–13.75 pg/ml, I2 = 100%, P CI = 0.22–6.42 pg/ml, I2 = 88%, P = 0.04). The symmetry of the funnel plots was not obvious, and Egger’s test showed that there was no significant small study effect in all groups. Conclusions This study showed that increased IL-6 correlates with long COVID-19. Such an informative revelation suggests IL-6 as a basic determinant to predict long COVID-19 or at least inform on the “early stage” of long COVID-19.

AIM: To evaluate the outcomes and elucidate the failure factors for trabeculectomy with mitomycin C (MMC) in Southwest Chinese patients. METHODS: A retrospective correlational study was conducted on the glaucomatous patients who underwent initial trabeculectomy with MMC in Southwest Hospital and had been followed up for 1-3y. A complete success for surgery is defined as a postoperative intraocular pressure (IOP) >5 and ≤21 mm Hg and 20% reduction of IOP compared to preoperative, without IOP-lowering medications. A qualified success for surgery is defined as the abovementioned postoperative IOP with or without IOP-lowering medications. The primary outcomes were IOP, the number of IOP-lowering medications, and cumulative success rate. The secondary outcomes included best corrected visual acuity (BCVA), mean deviation (MD) of visual field, major complications, and risk factors for surgical failure. RESULTS: A total of 325 eyes of 261 glaucomatous patients had been included in our study. Both the mean IOP and the number of IOP-lowering medications were significantly decreased from 32.9±12.0 to 16.4±5.7 mm Hg (P

An extended Roy equation including a bound state pole is used to study $\pi\pi$ scatterings at unphysical large pion masses when $\sigma$ becomes a bound state in one situation and stays as a broad resonance in the other case. The coupled integral equations at large pion masses are solved by taking the lattice driving terms and the Regge amplitudes as inputs. Relying on the solutions of Roy equations that respect unitarity, analyticity and crossing symmetry, we give predictions to the phase shifts with $IJ=00,11,20$ in the elastic energy region. We then perform analytic continuation into the complex $s$ plane to search for various poles, all of which are inside the validity domain of Roy equation. This is the first time that lattice data at unphysical large pion masses are analyzed within the rigorous Roy equation method., Comment: 20 pages, 13 figures, 4 tables, minor improvements, further clarification and interpretation, refs added, conclusions unmodified

Background: Poor prosthesis alignment during total knee arthroplasty could cause problems such as polyethylene spacer wear, leading to surgical failure and revision surgery. The problems caused by the malalignment of the tibial plateau prosthesis in the medial and lateral planes are unclear. We aimed to investigate the stress distribution and micromotion of the tibia when the tibial plateau prosthesis is translated 1 and 2 mm medially and laterally, respectively, using finite element analysis (FEA).Method: A non-homogeneous tibia model was created and load conditions when standing on two legs were applied using FEA to simulate the misaligned prosthesis. The stresses, stress distribution, and micromotion of the proximal tibia were analyzed in five positions of the tibial plateau prosthesis: Lateral-2 mm; Lateral-1 mm; Medium; Medial-2 mm; Medial-1 mm.Result: The maximum stress in the five groups with different misalignments of the platform was 47.29 MPa (Lateral-2 mm). The maximum micromotion among the five groups in different positions was 7.215 μm (Lateral-2 mm).Conclusion: When placing the tibial plateau prosthesis during total knee arthroplasty, an error of 2 mm or less is acceptable as long as it does not overhang.

Although genome-wide association studies are widely used to mine genes for quantitative traits, the effects to be estimated are confounded, and the methodologies for detecting interactions are imperfect. To address these issues, the mixed model proposed here first estimates the genotypic effects for AA, Aa, and aa, and the genotypic polygenic background replaces additive and dominance polygenic backgrounds. Then, the estimated genotypic effects are partitioned into additive and dominance effects using a one-way analysis of variance model. This strategy was further expanded to cover QTN-by-environment interactions (QEIs) and QTN-by-QTN interactions (QQIs) using the same mixed-model framework. Thus, a three-variance-component mixed model was integrated with our multi-locus random-SNP-effect mixed linear model (mrMLM) method to establish a new methodological framework, 3VmrMLM, that detects all types of loci and estimates their effects. In Monte Carlo studies, 3VmrMLM correctly detected all types of loci and almost unbiasedly estimated their effects, with high powers and accuracies and a low false positive rate. In re-analyses of 10 traits in 1439 rice hybrids, detection of 269 known genes, 45 known gene-by-environment interactions, and 20 known gene-by-gene interactions strongly validated 3VmrMLM. Further analyses of known genes showed more small (67.49%), minor-allele-frequency (35.52%), and pleiotropic (30.54%) genes, with higher repeatability across datasets (54.36%) and more dominance loci. In addition, a heteroscedasticity mixed model in multiple environments and dimension reduction methods in quite a number of environments were developed to detect QEIs, and variable selection under a polygenic background was proposed for QQI detection. This study provides a new approach for revealing the genetic architecture of quantitative traits.

Rohdea medogensis, a new species from Tibet, China, is described and illustrated. This species is similar to R. verruculosa in sharing characters of the perianth with ringed, fleshy appendage in throat and lobes abaxially with verruculose, but differs mainly by bracts 3 per flower, with several sterile bracts apically; outer bract ca. 4.5 cm long, ca. 3 mm broad; perianth globular; lobes shorter ca. 2.0 mm long, margin lacerated, apical subacut; the top of annular appendage margin lacerated, 2 mm above the top of perianth tube; stigma bend, without lobed; style conspicuous; ovary globose or subglobose, askew; berry and seed ellipsoid, askew.

The extremely high transmission rate of SARS-CoV-2 and severe cases of COVID-19 pose the two critical challenges in the battle against COVID-19. Increasing evidence has shown that the viral spike (S) protein-driven syncytia may be responsible for these two events. Intensive attention has thus been devoted to seeking S-guided syncytium inhibitors. However, the current screening campaigns mainly rely on either live virus-based or plasmid-based method, which are always greatly limited by the shortage of high-level biosafety BSL-3 facilities or too much labour-intensive work. Here, we constructed a new hybrid VEEV-SARS-CoV-2-S-eGFP reporter vector through replacement of the structural genes of Venezuelan equine encephalitis virus (VEEV) with the S protein of SARS-CoV-2 as the single structural protein. VEEV-SARS-CoV-2-S-eGFP can propagate steadily through cell-to-cell transmission pathway in S- and ACE2-dependent manner, forming GFP positive syncytia. In addition, a significant dose-dependent decay in GFP signals was observed in VEEV-SARS-CoV-2-S-eGFP replicating cells upon treatment with SARS-CoV-2 antiserum or entry inhibitors, providing further evidence that VEEV-SARS-CoV-2-S-eGFP system is highly sensitive to characterize the anti-syncytium-formation activity of antiviral agents. More importantly, the assay is able to be performed in a BSL-2 laboratory without manipulation of live SARS-CoV-2. Taken together, our work establishes a more convenient and efficient VEEV-SARS-CoV-2-S-eGFP replicating cells-based method for rapid screening of inhibitors blocking syncytium formation.

Di Tang,1,* Duo Zhang,1,* Yu Heng,1 Xiao-Ke Zhu,1 Han-Qing Lin,1 Jian Zhou,1 Lei Tao,1 Li-Ming Lu2 1Department of Otorhinolaryngology and ENT Institute, Eye & ENT Hospital, Fudan University, Shanghai, People’s Republic of China; 2Shanghai Institute of Immunology, Shanghai Jiaotong University School of Medicine, Shanghai, People’s Republic of China*These authors contributed equally to this workCorrespondence: Li-Ming Lu, Shanghai Institute of Immunology, Shanghai Jiaotong University School of Medicine, 280 South Chongqing Road, Shanghai, 200025, People’s Republic of China, Tel +86-13916235624, Fax +86-021-63846383, Email lulunew2003@163.com Lei Tao, ENT Institute and Department of Otorhinolaryngology, Eye & ENT Hospital, Fudan University, 83 Fenyang Road, Shanghai, 200031, People’s Republic of China, Tel +86-13916944810, Email doctortaolei@163.comPurpose: Chronic inflammation contributes to tumor initiation, progression, and immune escape. Neutrophils are the major component of inflammatory response and participate in the tumorigenesis process. However, compared to other immune cells in the tumor microenvironment of laryngeal squamous cell carcinoma (LSCC), neutrophils, especially the tumor-associated neutrophils (TANs), have not yet been comprehensively explored. The mechanism for regulating the crosstalk between TANs and tumor cells still remains unclear.Materials and Methods: The distribution profiles and phenotypic features of neutrophils and other inflammatory immune cell populations from a large LSCC patient cohort were systemically analyzed. Co-culturing of peripheral blood associated neutrophils (PANs) and TANs with PBMCs was performed, and the immunosuppression effect on T-cells was examined.Results: LSCC microenvironment is highly inflammatory with remarkable TANs infiltration, which is often associated with unfavorable prognosis and advanced clinical stage. We find that TANs in LSCC display morphologically immature and lower apoptosis, exhibit distinctively immunosuppressive phenotype of high PD-L1, and suppress CD8+ T lymphocytes proliferation and activation. We subsequently discover that PD-L1+TANs induced by LSCC-derived GM-CSF potently impair CD8+ T-cells proliferation and cytokines production function, which are partially blocked by a PD-L1-neutralizing antibody. Clinical data further support GM-CSF as an unfavorable prognostic biomarker and reveal a potential association with inflammatory immune cell infiltration, in particular neutrophils.Conclusion: Tumor-infiltrating PD-L1+ neutrophils induced by LSCC-derived GM-CSF suppress T cell proliferation and activation in the inflammatory microenvironment of LSCC and predict unfavorable prognosis. These TANs cripple antitumor T cell immunity and promote tumor progression. Our findings provide a basis for targeting PD-L1+TANs or GM-CSF as a new immunotherapeutic strategy for LSCC.Keywords: laryngeal squamous cell carcinoma, LSCC, tumor-associated neutrophils, TANs, PD-L1, inflammatory microenvironment, immune suppression, granulocyte-macrophage colony stimulating factor, GM-CSF

Objective Hemostasis markers associated with tumors have been widely studied. However, the associations between the coagulation factors and papillary thyroid carcinoma (PTC) prognosis remain unclear. The aim of the present study was to investigate these associations. Patients and methods PTC patients treated at Shengjing Hospital between 2013 and 2019 were retrospectively analyzed and divided into three groups. Clinical, ultrasound, and pathological patient characteristics were recorded. The blood routine, coagulation factors, and thyroid function data were compared. Results A total of 50 patients were selected and divided into Group 1 [complicated with second primary malignancies (SPMs)], Group 2 (with postoperative cervical lymph node metastasis), and Group 3 (control group). The ages of patients in Group 2 were significantly higher than those in the control group. The neutrophil ratio (%) in Group 1 was significantly higher than that in Groups 2 and 3, while its lymphocyte ratio (%) was significantly lower. The coagulation factor activated partial thromboplastin time (APTT) in the first and second groups was statistically significantly lower than that in the control group. There were no statistical differences in APTT between the first and second groups. Shorter APTT was associated with SPM and postoperative cervical lymph node metastasis. Conclusions Coagulation indicators, especially APTT, may be a new biomarker for predicting PTC prognosis and may provide a new molecular target, especially in combination with SPM and postoperative cervical lymph node metastasis.