Abstract Replication-competent coronaviruses carrying fluorescent protein-tagged structural proteins remain scarce. Using the highly attenuated pangolin coronavirus GX_P2V(short_3UTR) as a backbone, we generated GX_P2V-mBJ-N, a recombinant coronavirus in which the bright green fluorescent protein mBaoJin is fused to the nucleocapsid (N) protein. The reporter virus is attenuated and genetically unstable in normal Vero cells but can be amplified to high titers in cells expressing wild-type N, and its fluorescence directly reports N protein expression. Using this authentic-virus platform, we show that high-titer GX_P2V cross-neutralizing antibodies persist in most healthy individuals and that cepharanthine potently blocks viral entry. GX_P2V-mBJ-N thus provides a simple and reliable tool for coronavirus tracing, immune surveillance, and antiviral drug evaluation.

Purpose:Glucocorticoids are frequently administered in cases of severe drug-induced liver injury (DILI) to promote patient recovery and shorten hospitalization duration. However, their use is associated with an increased risk of infection. This study developed a predictive model for infection after glucocorticoid therapy in patients with DILI. Patients and Methods:We retrospectively analyzed patients with severe DILI treated with glucocorticoids at the Fifth Medical Center of the Chinese People's Liberation Army between 2017 and 2024. We constructed and interpreted eight machine learning models: random forest, support vector machine, generalized linear model, gradient boosting machine, least absolute shrinkage and selection operator, XGBoost, K-nearest neighbor classification, and artificial neural network. Decision curve analysis, calibration curves, receiver operating characteristics (ROC), and Shapley Additive Explanations model scores were used to interpret the optimal model. Results:Among the eight models, the gradient boosting machine showed the best results (area under the ROC curve: 0.981 and 0.928 for the validation and test sets, respectively) and had the smallest residuals. Decision curve analysis and calibration curves confirmed the model's strong clinical prediction performance. Globulin (GLO) was a key variable in the models, with significantly low levels in infected patients compared with those in the control group (p < 0.001). Patients with pre-treatment GLO levels below 20 g/L had a higher infection rate (41.1%), while those with post-treatment GLO levels below 21.5 g/L exhibited an even greater infection rate (82.3%). Conclusion:Our early warning model for the prediction of infection is valuable for guiding hormonal therapy for severe DILI. Monitoring changes in GLO levels may provide a simple and effective clinical monitoring tool for preventing infection development.

ABSTRACT Background Malaria remains a global health challenge, with 249 million cases and 608,000 deaths reported in 2022. While China achieved malaria elimination, imported cases surged by 194.4% in 2023, underscoring the need for rapid diagnostics. Traditional methods like microscopy and rapid diagnostic tests (RDTs) face limitations in sensitivity and infrastructure requirements. This study aimed to establish and optimize a “one‐pot” enzymatic recombinase amplification (ERA) assay for the molecular detection of Plasmodium falciparum and Plasmodium vivax, and to evaluate the efficacy of this assay through methodological verification and clinical performance. Methods We designed a specific ERA assay targeting the conserved regions of P. falciparum and P. vivax genetic material. We evaluated the sensitivity and specificity of this assay using synthetic plasmids and genomic material. Additionally, we tested the stability of the reaction by incorporating potential interfering substances into the reaction system. Finally, we analyzed the detection performance of the ERA method against real‐time fluorescent quantitative PCR and rapid diagnostic tests using clinical samples. Results The detection process could be completed within 25 min at 35°C–40°C, and the results could be interpreted either under UV light or using a GeneScope instrument. The detection limit of the ERA assay was 250 copies/mL, which was 40 times more sensitive than fluorescent quantitative PCR. When evaluating the clinical performance using 75 clinical specimens, the detection rate of the ERA method was 94.54% compared with 89.09% for fluorescent quantitative PCR. The ERA assay and fluorescent quantitative PCR can achieve positive detection when blood samples were diluted 1024 times or even 4096 times. Comparatively, the detection capabilities of rapid diagnostic tests were significantly lower than that of the ERA assay. Conclusion The ERA method shows good performance in the detection of P. falciparum and P. vivax, and can be used as a complementary tool for malaria screening and clinical diagnosis.

A significant number of infectious diseases affecting humans have been associated with zoonotic viruses. Wild small mammals, such as bats, rodents, and shrews, serve as natural reservoirs for a multitude of zoonotic viruses, particularly in Africa, where zoonosis is prevalent. Nevertheless, our knowledge of the virome composition within these hosts remains limited, impeding a more profound understanding of spillover events into human populations. We employed a viral metagenomics approach to characterize the virome in 846 wild small mammals sampled from Sierra Leone. Based on the complete RNA-dependent RNA polymerase genome, a total of 39 RNA viruses infecting mammals were identified, comprising 13 known viruses and 26 novel viruses. Notably, the Paramyxoviridae family exhibited the highest diversity of viral species across all three orders of wild mammal. The animal species Hipposideros jonesi and Lophuromys chrysopus were found to harbor the highest viral richness. Among these viral species, 15 were identified as cross-species transmitted viruses shared among different animal species, 3 were classified as zoonotic (Encephalomyocarditis virus, Rocahepevirus sp., and Lassa virus), while 3 others posed a potential risk for spillover (melian virus, Rodent hepacivirus, Hunnivirus A). Cross-species transmission analysis revealed that rodents played central roles in virus sharing, while cross-order viral transmission was less likely to occur in bats. Among 26 newly identified viruses, four viruses (Bat ledantevirus 2, Rattus rattus jeilongvirus, Miniopterus inflatus ribovirus, and Rat mamastrovirus) were predicted to have high zoonotic potential. Among them, Bat ledantevirus 2 exhibited the highest zoonotic potential and phylogenetic relatedness to the known human-infecting virus (Le Dantec virus). Further seroepidemiological analysis in patients, using single-round infectious virus particles as antigens, revealed the presence of neutralizing antibodies against Bat ledantevirus 2, a novel virus belonging to the Rhabdoviridae family. These findings highlight the critical need for enhanced surveillance at the human-animal interface in order to identify viruses with cross-species transmission potential prior to their spillover into human population.

Background: In many instances, Recurrent Spontaneous Abortion (RSA) has unexplained etiology and does not have any prognostic clues. Antioxidant vitamins are closely related to a series of diseases, including RSA. However, the function and underlying mechanism of calcitriol in the treatment of RSA has not been described. Materials and Methods: The pregnant mice were injected intraperitoneally with Lipopolysaccharide (LPS) to induce abortion. Calcitriol was used for the treatment and to avoid RSA. Pathological analysis was done to assess the abnormalities during RSA and its treatment using calcitriol. Results: After LPS administration, the mice exhibited high embryo absorption and pathological alterations, as well as, presented an increase in inflammation and apoptosis. Moreover, higher levels of NF-kappa B and inflammatory cytokines (TNF and IL-6) were noticed in serum and in isolated macrophages. Furthermore, we found alterations in the Nrf2 levels, enzymatic antioxidants and non-enzymatic antioxidants in serum. All these abnormalities were attenuated during calcitriol treatment in the appropriate groups of mice. Conclusion: Our results indicate that calcitriol may be involved in the modulation of inflammatory signaling and NF-kappa B pathway-mediated inflammation in RSA. Therefore, calcitriol could serve as a better prognostic regimen in the RSA treatment.

Background: Elderly patients infected with SARS-CoV-2 are at higher risk of developing cytokine storm and severe outcomes; however, specific immunological and proteomic biomarkers for early prediction remain unclear in this vulnerable group. Methods: We enrolled 182 elderly COVID-19 patients from the Chinese PLA General Hospital between November 2022 and April 2023, categorizing them based on progression to respiratory failure requiring mechanical ventilation (defined as severe progression). Olink proteomic analysis was performed on admission serum from 40 propensity score-matched samples, with differentially expressed proteins (DEPs) validated by cytometric bead array (CBA) in 178 patients. To predict severe progression, a model was developed using a 70% training set and validated on a 30% validation set. LASSO regression screened features followed by logistic regression and receiver operating characteristic (ROC) analysis to optimize the model by incrementally incorporating features ranked by random forest importance. Results: Elderly patients progressing to severe COVID-19 exhibited early immune dysregulation, including neutrophilia, lymphopenia, monocytopenia, elevated procalcitonin (PCT), C-reactive protein (CRP), interleukin-6 (IL-6), neutrophil-to-lymphocyte ratio (NLR), and systemic immune-inflammation index (SII), as well as coagulation dysfunction and multi-organ injury. Proteomics identified a set of biomarkers, including tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), and revealed disruptions in signaling pathways, including the mTOR and VEGF signaling pathways. The optimal predictive model, which incorporated PCT, IL-6, monocyte percentage, lymphocyte count, and TRAIL, achieved an area under curve (AUC) of 0.870 (0.729-1.000) during validation. TRAIL levels negatively correlated with fibrinogen (p < 0.05). Conclusions: Elderly COVID-19 patients with severe progression demonstrate early immune dysregulation, hyperinflammation, coagulation dysfunction, and multi-organ injury. The model we proposed effectively predicts disease progression in elderly COVID-19 patients, providing potential biomarkers for early clinical risk stratification in this vulnerable population.

Elderly individuals infected with SARS-CoV-2 are at higher risk of developing cytokine storms and severe outcomes, yet specific biomarkers remain unclear. In this study, we investigated the alteration of primary bile acid metabolism in elderly patients with severe COVID-19 using untargeted metabolomics (n = 31), followed by targeted metabolomics to compare patients with disease progression (n = 16) to those without (n = 48). Significant reductions in chenodeoxycholic acid (CDCA) and glycochenodeoxycholic acid (GCDCA) levels were identified in severe cases, with GCDCA levels at admission correlating strongly with peak inflammatory markers. In vitro, CDCA, GCDCA, and their receptors, Farnesoid X Receptor (FXR) and Takeda G-protein-coupled receptor 5 (TGR5), effectively inhibited the inflammatory response induced by SARS-CoV-2. NOD-like receptor pathway, activated by SARS-CoV-2, may modulate inflammatory cytokines under the treatment of CDCA, GCDCA, and TGR5. CDCA and GCDCA levels at admission predicted disease progression, suggesting their potential as biomarkers for severe COVID-19 in the elderly and highlighting their regulatory role in inflammation, pointing to new therapeutic avenues.

Elderly individuals represent a population at disproportionate risk for severe outcomes following SARS-CoV-2 infection. The humoral immune response critically controls viral replication and disease progression. However, the antibody responses and B-cell subsets to mutant strains of SARS-CoV-2 in elderly patients have not yet been fully elucidated. This study aims to elucidate the humoral immune response and B-cell subsets distribution in elderly patients infected with SARS-CoV-2 Omicron variant, thereby providing insights for identifying prognostic biomarkers and developing therapeutic strategies. Using enzyme-linked immunosorbent assays and pseudotyped virus neutralization assays, we determined plasma levels of RBD-specific IgA, IgM, IgG, and neutralizing antibodies from 46 elderly patients with SARS-CoV-2 Omicron variant infection within the first two weeks post-symptom onset (PSO). Using a multicolor flow cytometry approach, we analyze the frequencies of different B-cell subsets and assess the functional characterization of B-cells. In elderly non-severe patients, SARS-CoV-2 RBD-specific antibody levels (IgA, IgM, and IgG) increased progressively within the first two weeks post-symptom onset (PSO). In contrast, severe patients exhibited lower initial antibody levels during 0-3 days PSO but experienced a transient surge (11-fold for IgA, 12-fold for IgM, and 48-fold for IgG) during 4-7 days PSO, which was followed by a decline between 8-14 days. Despite this early elevated response in severe patients, both groups ultimately demonstrated generally weak neutralizing activity against the SARS-CoV-2 Omicron variant. Both elderly patient groups exhibited an higher proportion of plasmablasts (PB). Among these, class-switched IgM-IgG+ PB were significantly more abundant than IgM+IgG-PB. Notably, elderly severe patients showed a further lower in class-unswitched IgM+IgG-PB. Concurrently, IgA expression on PB was upregulated during early disease in all elderly patients. Beyond PB changes, IgG+double negative B (DNB) cells were higher than IgM+DNB cells in both groups. however, severe patients demonstrated a significant reduction in IgG+DNB cells frequencies. Furthermore, these elderly severe patients also exhibited a decline in co-stimulatory molecule expression (HLA-DR+CD80+) within both naive B (NB) and DNB cells, indicating a dysregulated humoral immune response. Elderly individuals generate RBD-specific and neutralizing antibody responses after SARS-CoV-2 Omicron variant infection, which correlate significantly with disease severity and infection duration. Clinically, these findings highlight the potential utility of antibody kinetics as prognostic biomarkers for stratifying elderly patients at high risk of severe outcomes, thereby informing tailored intervention strategies and vaccine booster optimization. Furthermore, SARS-CoV-2 Omicron variant infection affects the distribution of B-cell subsets, which may predict the long-term immune reconstitution capacity post-recovery. Notably, elderly patients exhibit pronounced PB expansion, a response pattern that facilitates antibody class-switching to enhance antiviral immunity. Such expansion represents a key immunologic feature in elderly patients with SARS-CoV-2 Omicron variant infection.

OBJECTIVE:Human adenovirus (HAdV) infection is common and can develop to serious conditions with high mortality, yet the mechanism of HAdV infection remains unclear. In the present study, the serum metabolite profiles of HAdV-7-infected patients with pneumonia or upper respiratory tract infection (URTI) were explored.METHODS:In total, 35 patients were enrolled in the study following an outbreak of HAdV-7 in the army, of whom 14 had pneumonia and 21 had URTI. Blood samples were collected at the acute stage and at the recovery stage and were analyzed by untargeted metabolomics.RESULTS:Over 90% of the differential metabolites identified between the pneumonia patients and URTI patients were lipids and lipid-like molecules, including glycerophospholipids, fatty acyls, and sphingolipids. The metabolic pathways that were significantly enriched were primarily the lipid metabolism pathways, including sphingolipid metabolism, glycerophospholipid metabolism, and linoleic acid metabolism. The sphingolipid metabolism was identified as a significantly differential pathway between the pneumonia patients and URTI patients and between the acute and recovery stages for the pneumonia patients, but not between the acute and recovery stages for the URTI patients. Ceramide and lactosylceramide, involved in sphingolipid metabolism, were significantly higher in the pneumonia patients than in the URTI patients with good discrimination abilities [area under curve (AUC) 0.742 and 0.716, respectively; combination AUC 0.801].CONCLUSION:Our results suggested that HAdV modulated lipid metabolism for both the patients with URTI and pneumonia, especially the sphingolipid metabolism involving ceramide and lactosylceramide, which might thus be a potential intervention target in the treatment of HAdV infection.

Background Galectin-9 (Gal-9) expression in patients with acute-on-chronic liver failure or its correlation with prognosis remain unclear. This study investigated the relationship between the prognosis of liver failure and the analysis of Gal-9 expression in patients with acute-on-chronic liver failure. Methods Patients with acute-on-chronic liver failure attributable to hepatitis B and those with chronic hepatitis B were included in this single-center prospective cohort study. The expressions of Gal-9 and Tim-3 in T cells were measured; the amounts of Gal-9 and related components in each group were examined using the enzyme-linked immunosorbent assay and flow cytometry. An investigation of the expression and distribution of Gal-9 in liver tissues was conducted using immunohistochemistry and immunofluorescence staining. Results The Gal-9 levels in the acute-on-chronic liver failure group were significantly higher than those in the chronic hepatitis B group. There was an upregulation of Gal-9 and T-cell immunoglobulin domain and mucin domain-3 expressions in peripheral blood T cells. Gal-9 was localized in the regenerative areas of liver tissues in patients with acute-on-chronic liver failure, co-localizing with Kupffer cells. Kaplan–Meier survival curves showed that patients with Gal-9 levels less than 9.6 ng/mL had a worse prognosis, with the area under the receiver operating characteristic curve being superior to that of the Model for End-Stage Liver Disease score. Conclusions Plasma Gal-9 levels can serve as a prognostic marker in patients with hepatitis B virus-acute-on-chronic liver failure. The predominant expression of Gal-9 in liver Kupffer cells suggests a potential immunosuppressive role in this patient population.

AbstractGalectin-9 (Gal-9) expression in patients with acute-on-chronic liver failure and its correlation with prognosis remain unclear. This study investigated the relationship between liver failure prognosis and Gal-9 expression analysis in patients with acute-on-chronic liver failure. Patients with acute-on-chronic liver failure attributable to hepatitis B and those with chronic hepatitis B were included in this single-center prospective cohort study. The Gal-9 levels in the acute-on-chronic liver failure group were significantly higher than those in the chronic hepatitis B group, and there was an upregulation of Gal-9 and T-cell immunoglobulin domain and mucin domain-3 expressions in peripheral blood T cells. Gal-9 was localized in the regenerative areas of liver tissues in patients with acute-on-chronic liver failure, co-localizing with Kupffer cells. Kaplan–Meier survival curves showed that patients with Gal-9 levels < 9.6 ng/ml had a worse prognosis, with the area under the receiver operating characteristic curve (AUC-ROC) being similar to that of the Model for End-Stage Liver Disease score. The combined ROC curve of the two had better predictive performance, with an AUC of 0.945. High Gal-9 levels in liver regenerative areas can serve as a prognostic marker, indicating a better prognosis for patients with hepatitis B virus-acute-on-chronic liver failure.

ABSTRACT Both human and non-human simian adenoviruses (HAdVs and SAdVs, respectively) have been used as gene therapy and vaccine vectors. The high prevalence of HAdVs and the neutralizing antibodies associated with prior infection, may limit HAdV-based vector use in human subjects. To overcome this drawback, a vector derived from a newly isolated and characterized macaque adenovirus was constructed. SAdVs (33.9%) were screened from 115 SAdV fecal samples collected at a zoological park. One novel SAdV was isolated and the whole genome was sequenced and analyzed. The pre-existing neutralizing antibody levels were very low against this isolate (10%). Interestingly, SAdV vector constructs that lack E3 region could not produce infectious progeny in HEK293 cells, suggesting that the E3 region is necessary for SAdV replication. The absence of E3 region could be compensated for by replacement with HAdV-5 E4orf6; the resultant construct could replicate well in HEK293 cells. The enhanced Green Fluorescent Protein (eGFP) was inserted into SAdV E3 region and expressed at high level. One-step growth curve showed that the replication of the SAdVs with HAdV-5 E4orf6 substitution and E1/E3 deletion was similar to that of wild-type SAdVs in HEK293 cells, but the modified SAdVs were replication-deficient in A549 cells which lack HAdV-5 E1A and E1B. Finally, we demonstrated that GZ3-12 could infect cells expressing hCAR or hDSG2 receptors. The successful isolation, characterization, and modification of novel SAdVs provide a potentially important vaccine and gene therapy candidate and a new strategy for the rapid acquisition and development of non-HAdV-based alternative vectors for human health applications. IMPORTANCE Adenoviruses are widely used in gene therapy and vaccine delivery. Due to the high prevalence of human adenoviruses (HAdVs), the pre-existing immunity against HAdVs in humans is common, which limits the wide and repetitive use of HAdV vectors. In contrast, the pre-existing immunity against simian adenoviruses (SAdVs) is low in humans. Therefore, we performed epidemiological investigations of SAdVs in simians and found that the SAdV prevalence was as high as 33.9%. The whole-genome sequencing and sequence analysis showed SAdV diversity and possible cross species transmission. One isolate with low level of pre-existing neutralizing antibodies in humans was used to construct replication-deficient SAdV vectors with E4orf6 substitution and E1/E3 deletion. Interestingly, we found that the E3 region plays a critical role in its replication in human cells, but the absence of this region could be compensated for by the E4orf6 from HAdV-5 and the E1 expression intrinsic to HEK293 cells.

Omicron BF.7 became the predominant SARS-CoV-2 variant in Beijing after the adjustment of COVID-19 response strategies in December 2022. The ability of antibodies elicited by BF.7 infection to cross-react with SARS-CoV-2-like viruses is unknown. This study aimed to investigate the cross-reactive neutralizing antibodies against SARS-CoV-2-related pangolin coronavirus GX_P2V in sera from vaccinated and/or SARS-CoV-2 infected individuals. All vaccinated individuals who recovered from Omicron BF.7 breakthrough infections exhibited substantially higher levels of neutralizing antibodies against GX_P2V among collected subject populations (geometric mean titer [GMT] = 362). Uninfected individuals who received four-mixed-dose vaccines also demonstrated higher levels of neutralizing antibodies (GMT = 44) against GX_P2V than those who received two- or three-dose vaccines and those who recovered from wild type SARS-CoV-2. This finding highlights the significance of prior and hybrid booster vaccinations with wild-type SARS-CoV-2 vaccines in generating potent cross-protective immunity against future spillovers of SARS-CoV-2-like viruses.

Abstract Harnessing SARS-CoV-2 related viruses from wild animals for live vaccines is an underexplored but promising approach, akin to the vaccinia virus used against smallpox. This study evaluated the potential of a live-attenuated vaccine derived from the pangolin coronavirus GX_P2V. We observed that the attenuated mutant, GX_P2V(short_3UTR), conferred significant and long-lasting protection against SARS-CoV-2 in golden hamsters, particularly after two doses. Furthermore, we show that sera from vaccinated convalescents of SARS-CoV-2 variant XBB.1.16 exhibited high titers of neutralizing antibody against GX_P2V(short_3UTR), suggesting GX_P2V(short_3UTR) shares neutralizing epitopes with XBB.1.16. We also show that the same sera from convalescents of XBB.1.16 displayed significantly reduced titers against the XBB.1.16 variant itself, suggesting a decoy-like immune evasion strategy that high-level cross-neutralizing antibodies induced by highly immunogenic epitopes of XBB.1.16 may not be able to efficiently neutralize the virus itself. Our findings emphasize GX_P2V(short_3UTR)'s potential as a broad-spectrum SARS-CoV-2 live vaccine and shed light on novel immune evasion mechanisms of the XBB.1.16 variant.

•The Delta variant patients exhaled ten million virus particles per hour.•87.14% of SARS-CoV-2 in exhaled breath was distributed in respiratory droplets.•Respiratory droplets may be a main transmission route of SARS-CoV-2 in exhaled breath.

Coronavirus disease 2019 (COVID-19) remains a serious global threat. The metabolic analysis had been successfully applied in the efforts to uncover the pathological mechanisms and biomarkers of disease severity. Here we performed a quasi-targeted metabolomic analysis on 56 COVID-19 patients from Sierra Leone in western Africa, revealing the metabolomic profiles and the association with disease severity, which was confirmed by the targeted metabolomic analysis of 19 pairs of COVID-19 patients. A meta-analysis was performed on published metabolic data of COVID-19 to verify our findings. Of the 596 identified metabolites, 58 showed significant differences between severe and nonsevere groups. The pathway enrichment of these differential metabolites revealed glutamine and glutamate metabolism as the most significant metabolic pathway (Impact = 0.5; -log10P = 1.959). Further targeted metabolic analysis revealed six metabolites with significant intergroup differences, with glutamine/glutamate ratio significantly associated with severe disease, negatively correlated with 10 clinical parameters and positively correlated with SPO2 (r(s) = 0.442, p = 0.005). Mini meta-analysis indicated elevated glutamate was related to increased risk of COVID-19 infection (pooled odd ratio [OR] = 2.02; 95% confidence interval [CI]: 1.17-3.50) and severe COVID-19 (pooled OR = 2.28; 95% CI: 1.14-4.56). In contrast, elevated glutamine related to decreased risk of infection and severe COVID-19, the pooled OR were 0.30 (95% CI: 0.20-0A4), and 0.44 (95% CI: 0.19-0.98), respectively. Glutamine and glutamate metabolism are associated with COVID-19 severity in multiple populations, which might confer potential therapeutic target of COVID-19, especially for severe patients.

目的 对西非疟疾病例进行实验室分析和确诊,探讨镜检、快速诊断试剂检测(RDT)、荧光定量PCR和巢式PCR在疟原虫检测中的应用效果.方法 选取西非驻塞拉利昂热带传染病预防和控制中心30例疑似感染疟原虫且均服用双氢青蒿素哌喹片患者血样,分别对其采取镜检、RDT、荧光定量PCR和巢式PCR法检测疟原虫并观察检验结果.结果 30例血样,镜检油镜下观察20例(66.66％)未见明显结构,10例(33.34％)可见恶性疟不同时期结构,其中恶性疟环状体4例(13.33％),裂殖体6例(20.00％).RDT检测28例阳性(93.33％),2例阴性(6.67％),阳性患者中T1阳性27例(90％),T1和T2同时阳性1例(3.33％).荧光定量PCR和巢式PCR检测结果,恶性疟阳性率100％,间日疟、三日疟、卵型疟均阴性.结论 镜检法灵敏度低于其他3种检测方法,RDT、荧光定量PCR和巢式PCR可获得较好的检测效果,且具有较好的临床适用性,但联合诊断的效果更佳,值得在临床推广实施.

Background: The coronavirus disease 2019 (COVID-19) is a highly infectious respiratory disease. There is no recommended antiviral treatment approved for COVID-19 in Sierra Leone, and supportive care and protection of vital organ function are performed for the patients. This study summarized the clinical characteristics, drug treatments, and risk factors for the severity and prognosis of COVID-19 in Sierra Leone to provide evidence for the treatment of COVID-19. Methods: Data of 180 adult COVID-19 patients from the 34th Military Hospital in Freetown Sierra Leone between March 31, 2020 and August 11, 2020 were retrospectively collected. Patients with severe and critically ill are classified in the severe group, while patients that presented asymptomatic, mild, and moderate disease were grouped in the non-severe group. The clinical and laboratory information was retrospectively collected to assess the risk factors and treatment strategies for severe COVID-19. Demographic information, travel history, clinical symptoms and signs, laboratory detection results, chest examination findings, therapeutics, and clinical outcomes were collected from each case file. Multivariate logistic analysis was adopted to identify the risk factors for deaths. Additionally, the clinical efficacy of dexamethasone treatment was investigated. Results: Seventy-six (42.22%) cases were confirmed with severe COVID-19, while 104 patients (57.78%) were divided into the non-severe group. Fever (56.67%, 102/180) and cough (50.00%, 90/180) were the common symptoms of COVID-19. The death rate was 18.89% (34/180), and severe pneumonia (44.12%, 15/34) and septic shock (23.53%, 8/34) represented the leading reasons for deaths. The older age population, a combination of hypertension and diabetes, the presence of pneumonia, and high levels of inflammatory markers were significantly associated with severity of COVID-19 development (P < 0.05 for all). Altered level of consciousness [odds ratio (OR) = 56.574, 95% confidence interval (CI) 5.645-566.940, P = 0.001], high levels of neutrophils (OR = 1.341, 95%CI 1.109-1.621, P = 0.002) and C-reactive protein (CRP) (OR = 1.014, 95%CI 1.003-1.025, P = 0.016) might be indicators for COVID-19 deaths. Dexamethasone treatment could reduce mortality [30.36% (17/56) vs. 50.00% (10/20)] among severe COVID-19 cases, but the results were not statistically significant (P > 0.05). Conclusions: The development and prognosis of COVID-19 may be significantly correlated with consciousness status, and the levels of neutrophils and CRP.

ObjectivesThe changes in metabolism by human adenovirus (HAdV) infection was unclear. The potential mechanism of HAdV-7 causing acute respiratory tract infection was explored.MethodsTotally 35 patients with HAdV-7 infection, 32 asymptomatic cases with HAdV-7 and 14 healthy controls were enrolled from an outbreak of HAdV-7 in the army. The serum samples were analyzed by untargeted and targeted metabolomics. The effects of differential metabolites were verified on HAdV-7 replication in an A549 cell line.ResultsThe untargeted metabolomics analysis revealed more significant changes in the classes of sphingolipids, polyketides, glycerolipids, fatty acyls, and carboxylic acids and their derivatives in the patients with HAdV-7 than in healthy controls. Two key metabolic pathways of secondary and primary bile acid biosynthesis were noted from pathway enrichment analysis. Targeted metabolomics analysis showed that the levels of unconjugated bile acids in the patients were significantly lower, while the levels of glyco- and tauro- conjugated bile acids in patients and asymptomatic cases were higher than those in the healthy controls. The profiles of cytokines and peripheral lymphocyte subsets obviously varied at different levels of bile acids, with significant differences after HAdV-7 infection. A cell verification test demonstrated that the replication of HAdV-7 significantly reduced when GCDCA and TCA were added.ConclusionBile acids inhibited HAdV-7 replication in vitro. Alterations in bile acids was metabolic signatures of HAdV-7 infected subjects, and our results suggested bile acids might play protective roles against HAdV-7 infection.

Human adenovirus (HAdV) infection causes excessive inflammation associated with severe tissue injury, such as pneumonia. The molecules involved in the underlying inflammatory mechanisms remain to be elucidated. Receptor for advanced glycation end product (RAGE) is mainly expressed on immune cells and lung tissues, and it is a key factor in the initiation and development of inflammation. RAGE can be cleaved by metalloprotease 9 (MMP9) to release the extracellular segment, which is named soluble RAGE (sRAGE), into the intercellular space, where it can bind to RAGE ligands and block RAGE activation and subsequent inflammation. In our study, we enrolled HAdV-infected patients and their contacts to examine the relationship between sRAGE and inflammation induced by HAdV infection. The results showed that HAdV infection stimulated inflammatory cytokine secretion, increased such as high mobility group box 1 (HMGB1) levels, and suppressed sRAGE expression. sRAGE levels were significantly different between patients with or without pneumonia. We also found that MMP9 was significantly lower in patients with pneumonia, and it was positively correlated with sRAGE levels over 7 days after disease onset. The mitogen-activated protein kinase (MAPK) pathway is an important immune activation signaling pathway that is regulated by RAGE. We observed the activation of the MAPK pathway in the peripheral blood mononuclear cells (PBMCs) of patients. Negative correlations between sRAGE and phosphorylated JNK and p38 were observed. These results suggest that sRAGE is involved in HAdV-induced inflammatory responses, and might be a potential therapeutic target to alleviate the HAdV-induced excessive inflammation.