Background: Influenza A virus (IAV) is a negative-sense RNA virus of the Orthomyxoviridae family and is the etiological agent of a highly contagious acute respiratory disease that can lead to acute lung injury. Objective: To elucidate the molecular mechanisms of IAV infection, an integrative research approach combining gene expression profiling, multinetwork analysis, and in vivo experimental validations was employed. Methods: First, a series of network-based analyses were performed, including protein-protein interaction network construction, weighted gene co-expression network analysis, and subsequent gene set enrichment analysis, to identify the major underlying mechanisms of IAV infection. Following gene expression analysis, core targets, both direct and indirect regulators, were screened. An IAV (H1N1) strain A/PR/8/34-induced acute lung injury mouse model was constructed for in vivo validations. Batch one included two groups to evaluate findings from the multi-network analysis: Mock (n = 10; 5 males and 5 females) and IAV (n = 10; 5 males and 5 females). Batch two included three groups to assess the role of toll-like receptor 3 (TLR3) in IAV infection: Mock (n = 6; 3 males and 3 females), IAV (n = 6; 3 males and 3 females), and TLR3 inhibitor (n = 6; 3 males and 3 females). Body weight was measured on days 0, 3, and 5 after infection. On day 5, lung tissues were collected to assess viral load and histopathological changes. Key targets were examined using enzyme-linked immunosorbent assay, Western blotting, and immunofluorescence staining, both in sera and lung tissues. Results: IAV infection was significantly associated with dysregulation of the immune-inflammation system, such as the LTR, nucle-otide-binding oligomerization domain-(NOD) like receptor, retinoic acid-inducible gene I-like receptor, and nuclear factor kappa-B signaling pathways. Gene set enrichment analysis further indicated that the TLR and necroptosis signaling pathways played crucial roles in the progression of IAV infection (TLR signaling pathway normalized enrichment score = 2.3941, P = 1.00 × 10 ⁻¹⁰; necroptosis normalized enrichment score = 1.9421, P = 6.21 × 10 ⁻⁷). Among the core targets, TLR3 and mixed lineage kinase domain-like protein (MLKL) may regulate gene expression at the transcriptional level (all P < 0.05). In vivo validation using an IAV (PR8) infected acute lung injury mouse model demonstrated increased viral load and lung index, alveolar structural damage, and inflammatory cell infiltration. Immunofluorescence staining exhibited large gaps in Lamin B1 staining and breaches in Emerin signals following IAV-PR8 infection. Expression levels of TLR3, p-receptor-interacting serine/threonine-protein kinase 3 (RIPK3)/RIPK3, and p-mixed lineage kinase domain-like protein (MLKL)/MLKL proteins in lung tissues, as well as proinflammatory factors and mediators in sera, were significantly elevated after IAV infection. Moreover, enhanced neutrophil infiltration (myeloperoxidase) and citrullinated histone H3 (a neutrophil extracellular trap-specific marker), both established indicators of neutrophil extracellular trap formation, were observed. Notably, treatment with a TLR3 inhibitor significantly ameliorated IAV-induced acute lung injury by regulating necroptosis-related targets. Conclusion: Our study provides network-based in vivo evidence that TLR3-receptor-interacting serine/threonine-protein kinase 3-MLKL-mediated necroptosis may underlie IAV-induced acute lung injury and could serve as a potential therapeutic target in severe influenza cases.

Objective To analyze the clinical characteristics of patients with coronary heart disease complicated by gastrointestinal hemorrhage and explore the independent prognostic factors.Methods The clinical data of patients with coronary heart disease complicated by gastrointestinal hemorrhage who were treated in the department of emergency of the First Affiliated Hospital of Zhejiang University School of Medicine from January 1,2022 to December 31,2023 were collected,including general information:gender,age,smoking history,drinking history,gastrointestinal hemorrhage-related symptoms(hematemesis,melena,hematochezia,hematemesis combined with melena)or positive fecal occult blood test,past medical history[hypertension,diabetes,atrial fibrillation,liver cirrhosis,malignancy,prior gastrointestinal hemorrhage],coronary heart disease treatment history(history of coronary stent implantation,history of coronary artery bypass grafting),history of antithrombotic drugs,vital signs on admission[body temperature,pulse rate,systolic blood pressure,pulse oxygen saturation(SpO2)],laboratory tests[random blood glucose,white blood cell count(WBC),hemoglobin(Hb),platelet count(PLT),C-reactive protein(CRP),serum creatinine(SCr),blood urea nitrogen(BUN),serum albumin(Alb),alanine aminotransferase(ALT),aspartate aminotransferase(AST),MB isoenzyme of creatine kinase(CK-MB),troponin I(TNI),brain natriuretic peptide(BNP),fibrinogen(Fib),prothrombin time(PT),activated partial thromboplastin time(APTT),D-dimer,ejection fraction(EF)],treatment process[gastrointestinal hemorrhage management(gastrointestinal endoscopy examination or treatment,vascular interventional therapy),blood transfusion volume(red blood cells,plasma),ICU admission rate and admission time,hospital stay],the causes of gastrointestinal hemorrhage comprehensively inferred from imaging and gastrointestinal endoscopy.According to the treatment outcome,the patients were divided into the cured group and the death group,and the differences in clinical data between the two groups were compared.Multivariate Logistic regression analysis was used to identify the independent influencing factors of death outcome.The receiver operating characteristic(ROC)curves were plotted to compare the predictive value of each factor for prognosis.Results A total of 526 patients were enrolled,including 486 in the cured group and 40 in the death group.The causes of death were multiple organ dysfunction syndrome(22 cases),acute myocardial infarction(7 cases),malignant arrhythmia(2 cases),respiratory failure(2 cases),infection(2 cases),hemorrhagic shock(3 cases),acute cerebral infarction(1 case),and cerebral hemorrhage(1 case).No statistically significant differences were found between the two groups in terms of gender,age,smoking history,excessive drinking history,symptoms(hematemesis,hematochezia,positive fecal occult blood test,hematemesis combined with melena),past medical history(hypertension,atrial fibrillation,liver cirrhosis,tumor history,prior gastrointestinal hemorrhage),coronary heart disease treatment history,antithrombotic drug history,body temperature,pulse rate,SpO2,random blood glucose,WBC,Hb,PLT,ALT,AST,CK-MB,TNI,Fib,APTT,EF,gastrointestinal hemorrhage management,causes of gastrointestinal hemorrhage,red blood cell/plasma transfusion volume,and hospital stay(all P>0.05).Compared with the cured group,the death group had a lower rate of melena as the initial symptom,systolic blood pressure,and serum Alb level[melena rate:32.50%(13/40)vs.66.26%(322/486),systolic blood pressure(mmHg,1 mmHg≈0.133 kPa):104.71±8.52 vs.122.52±22.59,Alb(g/L):30.80(27.60,33.70)vs.34.70(32.65,39.05),all P<0.05],and a higher prevalence of diabetes,higher levels of CRP,SCr,BUN,BNP,PT,D-dimer,a higher ICU admission rate,and a longer ICU stay[diabetes prevalence:70.00%(28/40)vs.29.42%(143/486),CRP(mg/L):94.91(30.69,125.56)vs.2.95(1.17,24.31),SCr(μmol/L):200.0(123.0,407.0)vs.82.5(66.8,112.0),BUN(mmol/L):15.81(14.00,22.21)vs.7.61(5.00,14.49),BNP(ng/L):2 970.50(1 504.25,6 193.50)vs.442.00(141.25,1 590.25),PT(s):13.50(12.60,22.50)vs.12.25(11.58,13.30),D-dimer(μg/L):5601(4115,11352)vs.609(267,1350),ICU admission rate:67.50%(27/40)vs.6.99%(34/486),ICU stay(days):3(2,16)vs.0(0,0),all P<0.05].With clinical outcomes(death,cure)as the dependent variable,variables with statistical significance in univariate analysis were included in the multivariate Logistic regression analysis.The results showed that systolic blood pressure and serum Alb were independent protective factors for death outcome[odds ratio(OR)=0.960,0.818;95%confidence interval(95%CI)was 0.933-0.987,0.719-0.932;P=0.004,0.002;respectively],while CRP and SCr were independent risk factors for death outcome[OR=1.013,1.004;95%CI was 1.006-1.020,1.002-1.006,all P<0.001].Taking death outcome as the state variable and serum Alb,systolic blood pressure,SCr,and CRP as the test variables respectively,the ROC curves were plotted.The results indicated that serum Alb,systolic blood pressure,SCr,CRP and their combined detection all had high predictive efficacy for the death outcome[area under the curve(AUC)=0.825,0.775,0.862,0.819,0.928;95%CI was 0.766-0.885,0.729-0.820,0.818-0.905,0.703-0.934,0.891-0.965;sensitivity was 78.6%,92.8%,85.7%,85.7%,92.9%;specificity was 72.7%,72.5%,78.8%,83.8%,77.8%,all P<0.001].The combined detection of the 4 factors had the highest predictive efficacy(all P<0.05).Conclusions Systolic blood pressure,serum Alb,CRP,and SCr were independent influencing factors for the in-hospital death outcome of patients with coronary heart disease complicated by gastrointestinal hemorrhage.The Logistic regression model constructed based on these factors had better predictive efficacy for death outcome than single factor analysis.

Objective To analyze changes in intestinal microbiota composition and metabolites in mice with nitrous oxide poisoning using 16S rDNA sequencing and metabolomics,and to examine correlations between gut microbes and metabolites in order to explore the mechanisms of nitrous oxide poisoning.Methods C57BL/6 mice were randomly divided into a control group and a nitrous oxide poisoning group(n=6).The poisoning group was exposed to 90,000 ppm nitrous oxide twice daily for 1 h over 28 days,while the control group was exposed to air.Fecal samples were collected 24 h after the last exposure.16S rDNA sequencing was used to analyze structural differences in microbial communities and identify significantly different taxa.Metabolomics analysis was performed to detect changes in fecal metabolites and identify differential metabolites.Correlation analysis was conducted between differential microbiota and metabolites.Results 16S rDNA sequencing showed that the poisoning group had increased microbial abundance compared with controls,while species diversity remained unchanged.Significant differences were observed in gut microbiota structure between groups.Metabolomics identified 112 differential metabolites related to nitrous oxide poisoning,mainly involving the cAMP signaling pathway and sphingolipid metabolism.Spearman correlation analysis revealed a strong association between differential microbiota and differential metabolites.Conclusion Nitrous oxide poisoning alters the structure and metabolic profiles of intestinal microbiota.Changes in microbial abundance affect multiple metabolic pathways,which may be related to damage to the nervous and hematological systems.These findings provide a basis for further research on the mechanisms of nitrous oxide poisoning and for clinical treatment.

Objective To analyze changes in intestinal microbiota composition and metabolites in mice with nitrous oxide poisoning using 16S rDNA sequencing and metabolomics,and to examine correlations between gut microbes and metabolites in order to explore the mechanisms of nitrous oxide poisoning.Methods C57BL/6 mice were randomly divided into a control group and a nitrous oxide poisoning group(n=6).The poisoning group was exposed to 90,000 ppm nitrous oxide twice daily for 1 h over 28 days,while the control group was exposed to air.Fecal samples were collected 24 h after the last exposure.16S rDNA sequencing was used to analyze structural differences in microbial communities and identify significantly different taxa.Metabolomics analysis was performed to detect changes in fecal metabolites and identify differential metabolites.Correlation analysis was conducted between differential microbiota and metabolites.Results 16S rDNA sequencing showed that the poisoning group had increased microbial abundance compared with controls,while species diversity remained unchanged.Significant differences were observed in gut microbiota structure between groups.Metabolomics identified 112 differential metabolites related to nitrous oxide poisoning,mainly involving the cAMP signaling pathway and sphingolipid metabolism.Spearman correlation analysis revealed a strong association between differential microbiota and differential metabolites.Conclusion Nitrous oxide poisoning alters the structure and metabolic profiles of intestinal microbiota.Changes in microbial abundance affect multiple metabolic pathways,which may be related to damage to the nervous and hematological systems.These findings provide a basis for further research on the mechanisms of nitrous oxide poisoning and for clinical treatment.

Objective To analyze the clinical characteristics of patients with coronary heart disease complicated by gastrointestinal hemorrhage and explore the independent prognostic factors.Methods The clinical data of patients with coronary heart disease complicated by gastrointestinal hemorrhage who were treated in the department of emergency of the First Affiliated Hospital of Zhejiang University School of Medicine from January 1,2022 to December 31,2023 were collected,including general information:gender,age,smoking history,drinking history,gastrointestinal hemorrhage-related symptoms(hematemesis,melena,hematochezia,hematemesis combined with melena)or positive fecal occult blood test,past medical history[hypertension,diabetes,atrial fibrillation,liver cirrhosis,malignancy,prior gastrointestinal hemorrhage],coronary heart disease treatment history(history of coronary stent implantation,history of coronary artery bypass grafting),history of antithrombotic drugs,vital signs on admission[body temperature,pulse rate,systolic blood pressure,pulse oxygen saturation(SpO2)],laboratory tests[random blood glucose,white blood cell count(WBC),hemoglobin(Hb),platelet count(PLT),C-reactive protein(CRP),serum creatinine(SCr),blood urea nitrogen(BUN),serum albumin(Alb),alanine aminotransferase(ALT),aspartate aminotransferase(AST),MB isoenzyme of creatine kinase(CK-MB),troponin I(TNI),brain natriuretic peptide(BNP),fibrinogen(Fib),prothrombin time(PT),activated partial thromboplastin time(APTT),D-dimer,ejection fraction(EF)],treatment process[gastrointestinal hemorrhage management(gastrointestinal endoscopy examination or treatment,vascular interventional therapy),blood transfusion volume(red blood cells,plasma),ICU admission rate and admission time,hospital stay],the causes of gastrointestinal hemorrhage comprehensively inferred from imaging and gastrointestinal endoscopy.According to the treatment outcome,the patients were divided into the cured group and the death group,and the differences in clinical data between the two groups were compared.Multivariate Logistic regression analysis was used to identify the independent influencing factors of death outcome.The receiver operating characteristic(ROC)curves were plotted to compare the predictive value of each factor for prognosis.Results A total of 526 patients were enrolled,including 486 in the cured group and 40 in the death group.The causes of death were multiple organ dysfunction syndrome(22 cases),acute myocardial infarction(7 cases),malignant arrhythmia(2 cases),respiratory failure(2 cases),infection(2 cases),hemorrhagic shock(3 cases),acute cerebral infarction(1 case),and cerebral hemorrhage(1 case).No statistically significant differences were found between the two groups in terms of gender,age,smoking history,excessive drinking history,symptoms(hematemesis,hematochezia,positive fecal occult blood test,hematemesis combined with melena),past medical history(hypertension,atrial fibrillation,liver cirrhosis,tumor history,prior gastrointestinal hemorrhage),coronary heart disease treatment history,antithrombotic drug history,body temperature,pulse rate,SpO2,random blood glucose,WBC,Hb,PLT,ALT,AST,CK-MB,TNI,Fib,APTT,EF,gastrointestinal hemorrhage management,causes of gastrointestinal hemorrhage,red blood cell/plasma transfusion volume,and hospital stay(all P>0.05).Compared with the cured group,the death group had a lower rate of melena as the initial symptom,systolic blood pressure,and serum Alb level[melena rate:32.50%(13/40)vs.66.26%(322/486),systolic blood pressure(mmHg,1 mmHg≈0.133 kPa):104.71±8.52 vs.122.52±22.59,Alb(g/L):30.80(27.60,33.70)vs.34.70(32.65,39.05),all P<0.05],and a higher prevalence of diabetes,higher levels of CRP,SCr,BUN,BNP,PT,D-dimer,a higher ICU admission rate,and a longer ICU stay[diabetes prevalence:70.00%(28/40)vs.29.42%(143/486),CRP(mg/L):94.91(30.69,125.56)vs.2.95(1.17,24.31),SCr(μmol/L):200.0(123.0,407.0)vs.82.5(66.8,112.0),BUN(mmol/L):15.81(14.00,22.21)vs.7.61(5.00,14.49),BNP(ng/L):2 970.50(1 504.25,6 193.50)vs.442.00(141.25,1 590.25),PT(s):13.50(12.60,22.50)vs.12.25(11.58,13.30),D-dimer(μg/L):5601(4115,11352)vs.609(267,1350),ICU admission rate:67.50%(27/40)vs.6.99%(34/486),ICU stay(days):3(2,16)vs.0(0,0),all P<0.05].With clinical outcomes(death,cure)as the dependent variable,variables with statistical significance in univariate analysis were included in the multivariate Logistic regression analysis.The results showed that systolic blood pressure and serum Alb were independent protective factors for death outcome[odds ratio(OR)=0.960,0.818;95%confidence interval(95%CI)was 0.933-0.987,0.719-0.932;P=0.004,0.002;respectively],while CRP and SCr were independent risk factors for death outcome[OR=1.013,1.004;95%CI was 1.006-1.020,1.002-1.006,all P<0.001].Taking death outcome as the state variable and serum Alb,systolic blood pressure,SCr,and CRP as the test variables respectively,the ROC curves were plotted.The results indicated that serum Alb,systolic blood pressure,SCr,CRP and their combined detection all had high predictive efficacy for the death outcome[area under the curve(AUC)=0.825,0.775,0.862,0.819,0.928;95%CI was 0.766-0.885,0.729-0.820,0.818-0.905,0.703-0.934,0.891-0.965;sensitivity was 78.6%,92.8%,85.7%,85.7%,92.9%;specificity was 72.7%,72.5%,78.8%,83.8%,77.8%,all P<0.001].The combined detection of the 4 factors had the highest predictive efficacy(all P<0.05).Conclusions Systolic blood pressure,serum Alb,CRP,and SCr were independent influencing factors for the in-hospital death outcome of patients with coronary heart disease complicated by gastrointestinal hemorrhage.The Logistic regression model constructed based on these factors had better predictive efficacy for death outcome than single factor analysis.

School-age children are in a specific development stage corresponding to juvenility, when the white matter of the brain experiences ongoing maturation. Diffusion-weighted magnetic resonance imaging (DWI), especially diffusion tensor imaging (DTI), is extensively used to characterize the maturation by assessing white matter properties in vivo. In the analysis of DWI data, spatial normalization is crucial for conducting inter-subject analyses or linking the individual space with the reference space. Using tensor-based registration with an appropriate diffusion tensor template presents high accuracy regarding spatial normalization. However, there is a lack of a standardized diffusion tensor template dedicated to school-age children with ongoing brain development. Here, we established the school-age children diffusion tensor (SACT) template by optimizing tensor reorientation on high-quality DTI data from a large sample of cognitively normal participants aged 6-12 years. With an age-balanced design, the SACT template represented the entire age range well by showing high similarity to the age-specific templates. Compared with the tensor template of adults, the SACT template revealed significantly higher spatial normalization accuracy and inter-subject coherence upon evaluation of subjects in two different datasets of school-age children. A practical application regarding the age associations with the normalized DTI-derived data was conducted to further compare the SACT template and the adult template. Although similar spatial patterns were found, the SACT template showed significant effects on the distributions of the statistical results, which may be related to the performance of spatial normalization. Looking forward, the SACT template could contribute to future studies of white matter development in both healthy and clinical populations. The SACT template is publicly available now ( https://figshare.com/articles/dataset/SACT_template/14071283 ).

Objective To investigate the changes in cognitive function in patients with early Parkinson's disease (PD) in a 5-year follow-up.Methods A total of 181 PD and 173 normal participants were recruited between January 2009 and January 2012 at the Department of Neurology,the Second Hospital of Jiaxing City.Regression analysis was used to evaluate the risk factors of cognitive impairment,and changes in subdomains of the Montreal Cognitive Assessment (MoCA) were compared annually with baseline data.Results Baseline clinical data were similar between the two groups.The level of cognitive impairment was positively correlated to the age of onset and the Hamilton Anxiety Scale (H AMD) (t =3.326,P＜ 0.05;t =5.211,P＜0.01),and negatively correlated to education level (t=-2.505,P＜0.05).There were no statistical differences in the first (26.5 ± 2.6),second (26.3±3.2) and third year (25.9±2.9) in the total scores of MoCA,which significantly increased in the forth (24.4 ± 2.3,P＜0.05) and fifth (24.1 ± 1.2,P＜0.05) year compared with baseline levels (26.7±2.9).However,in the control group,differences between any two years in total scores were not statistically significant (all P＞0.05).Seven subdomains of MoCA were attenuated,and four of them were significant different between the groups,including delayed recall,attention,abstraction task and orientation after the forth year of follow-up (all P ＜ 0.05).Meanwhile,visuospatial execution capacity was attenuated before the third year,and then rose markedly (P＜0.05).In the control group,the between-year differences of seven subdomains were not statistically significant (all P＞0.05).Conclusions Cognitive function of PD patients decreases significantly in some domains,such as delayed recall,attention,abstraction task and orientation after a five-year followup.

Objective To analyze the results of the assessment of malaria elimination in 8 counties(districts)in Huai'an City,Jiangsu Province,so as to provide evidences for consolidating the achievement of malaria elimination. Methods The da-ta from the network reports of malaria epidemic situation,blood examinations of febrile patients,epidemiological questionnaires of malaria cases,investigation and disposal of epidemic focuses were collected and analyzed in Huai'an City from 2010 to 2016. The results of the assessment on the clinicians'ability of malaria diagnosis and treatment as well as the inspectors'skill of Plas-modium microscopic examinations were analyzed. In the natural village where the last local case of malaria was reported ,no less than 200 blood filter papers were made for gene detection of Plasmodium. Results From 2010 to 2016,308 malaria cases were reported in Huai'an City. Totally 323002 patients received blood detection of malaria,among which 272 ones were positive and the positive rate was 0.08％. The majority of the malaria cases were imported,except 19 and 3 local infection cases in 2010 and 2011,respectively. Except Qinghe District,a total of 1420 blood filter papers from the other 7 counties(districts)were detect-ed genetically,and the results were all negative. Six of the eight counties(districts)gained an average score of 20 points on the Plasmodium microscopic examination,and Jinhu County and Huai'an District got a minimum average score of 19.6 points. In ca-pacity assessment of malaria diagnosis and treatment,Hongze District gained the highest average score of 20 points,and Huai'an District got the lowest average score of 18.8 points. The malaria elimination assessment scores of the eight counties(districts) were all above 97 points. All the 8 counties(districts)in Huai'an City passed the malaria elimination assessment at county(dis-trict)level until 2016. Conclusions All the 8 counties(districts)in Huai'an City have passed the malaria elimination assess-ment with high scores. After the malaria elimination,the monitoring still should continue to consolidate the achievement.

Objective To evaluate the cost-benefit of netting cultivation to block the spread of Oncomelania snails in lake re-gions. Methods The cost-benefit of netting cultivation was investigated by interviewing in the field,the cost of Oncomelania snail survey and control was investigated by retrospective review in Gaoyou Lake regions of Jinhu County from 2009 to 2011. The benefit of netting cultivation to block the spread of snails in lake regions was calculated by the benefit-cost ratio(BCR),and then the cost-benefit of them was calculated. Results The area of netting cultivation in Gaoyou Lake regions was 70.77 hm2,the aver-age cost of netting cultivation was 495 595 yuan every year,the average income was 962 000 yuan every year,and the average ben-efit of netting cultivation was 466 405 yuan. The average cost of Oncomelania snail survey and control from 2009 to 2011 was 85 047.87 yuan in Gaoyou Lake regions. The ratio of cost-benefit was 1.11∶1. The benefit was more than the cost. Conclusion There is a well benefit in lake regions during blocking the spread of snails by netting cultivation.