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.

ObjectiveTo understand the prevalence of tobacco use among residents aged 15 years old and above in Jiading District of Shanghai, and to provide a scientific basis for formulating effective regional tobacco control policies. MethodsThe Probability Proportional to Size (PPS) sampling method was employed to select one neighborhood committee (village) from each of Jiading District’s 11 subdistricts as survey monitoring sites. From the household registry of each selected neighborhood committee (village), 50 residential households were randomly sampled, resulting in a total of 550 households surveyed. Within each selected household, one permanent resident aged 15 years old or above was randomly chosen to complete the questionnaire. The content covered general information, exposure to secondhand smoke, awareness of tobacco related health hazards, etc. Statistical analyses were performed using SPSS 27.0 software. ResultsA total of 550 questionnaires were distributed, with 549 valid responses collected, and the effective response rate was 99.82%. The survey findings revealed that in 2023, the current smoking rate among individuals aged 15 years old and above in Jiading District of Shanghai was 18.76%, with males accounting for 38.93% and females accounting for 2.62%. Male gender was identified as a relative factor for smoking (OR=34.108, 95%CI: 14.440‒80.722), whereas higher education attainment emerged as a protective factor against smoking (OR=0.388, 95%CI: 0.184‒0.820). Among non-smokers aged 15 years old and above in Jiading District, the secondhand smoke exposure rate was 46.86%, with statistically significant differences observed across different age groups and occupations (P<0.001). Restaurants exhibited the highest secondhand smoke exposure rate of 42.14%, followed by households bearing (36.79%). The overall awareness rate among individuals aged 15 years old and above in Jiading District regarding four smoking-related diseases, namely stroke, heart disease, lung cancer, and impotence, was 48.45%, while the awareness rate for three secondhand smoke-induced diseases, namely adult heart disease, pediatric pulmonary disease, and adult lung cancer, was 64.29%. ConclusionThere is still room for reduction in the prevalence of tobacco use among individuals aged 15 yeas old and above in Jiading District. The exposure to secondhand smoke is severe, and residents have low awareness of the harm of tobacco. Tobacco control law enforcement should be strengthened, smoke-free-home health education should be included in tobacco control efforts, and targeted health education should be carried out.

Objective: To establish and validate a whole blood (WB) supply model, thereby providing practical experience for the clinical application of WB in domestic trauma emergency care and informing the development of a wartime blood supply system for the military. Methods: A “10×24” WB supply model was established by formulating blood collection protocols, storage standards, and transfusion criteria. Multiple WB samples were tested under specific storage conditions to assess key indicators at different time points, including red blood cell (RBC), white blood cell (WBC), and platelet counts, hemoglobin concentration, coagulation parameters (PT, APTT, TT, FIB), coagulation factor activity, thromboelastography (TEG) parameters, and electrolyte levels. Additionally, clinical data from hemorrhagic patients who met the criteria for WB transfusion and were admitted between March and July 2024 were analyzed to evaluate WB transfusion volume. Results: RBC counts and hemoglobin levels remained stable in WB stored at 4℃ for up to 10 days. However, platelet counts and coagulation function (PT, APTT) significantly declined with prolonged storage, while potassium levels increased. From March to July 2024, the model was successfully applied to 23 patients with acute hemorrhage, with a median WB transfusion volume of 543 mL. A detailed case study of a severe traumatic hemorrhagic shock patient was reported, who was successfully treated with 5.5 units of refrigerated WB combined with component blood. Conclusion: The “10×24” WB supply model demonstrated acceptable changes in critical quality parameters under strict management and a 10-day rotation cycle. This model effectively supports the treatment of acute hemorrhage and holds promise for integration into the future wartime blood supply system of the military.

Acute liver injury (ALI) serves as a critical precursor and major etiological factor in the progression and ultimate manifestation of various hepatic disorders. The prevention and treatment of ALI is still a serious global challenge. Given the limited therapeutic options for ALI, exploring novel targeted therapeutic agents becomes imperative. The potential therapeutic efficacy of inhibiting RIPK2 is highlighted, as it may provide significant benefits by attenuating the MAPK pathway and NF-κB signaling. Herein, we propose a CMD-OPT model, a two-stage molecular optimization tool for the rapid discovery of RIPK2 inhibitors with optimal properties. Compound RP20, which targets the ATP binding site, demonstrated excellent kinase specificity, ideal oral pharmacokinetics, and superior therapeutic effects in a model of APAP-induced ALI, positioning RP20 as a promising preclinical candidate. This marks the first application of RIPK2 inhibitors in ALI treatment, opening a novel therapeutic pathway for clinical applications. These results highlight the efficacy of the CMD-OPT model in producing lead compounds from known active molecules, showcasing its significant potential in drug discovery.

ObjectiveTo investigate the pathogenic spectrum and epidemiological characteristics of diarrheal disease in Fengxian District of Shanghai, and to provide scientific basis for the prevention and control of diarrheal diseases. MethodsBasic information of the initial adult cases visited diarrheal disease surveillance sentinel hospital in Fengxian District, Shanghai, was collected from August 2013 to 2023, and fecal samples were collected at 1∶5 sampling intervals to isolate and identify 5 kinds of diarrheagenic Escherichia coli (DEC), Salmonella (SAL), Vibrio parahaemolyticus, Campylobacter, Vibrio cholerae, Shigella and Yersinia enterocolitica (YE). Simultaneously, nucleic acid detection was performed for 3 kinds of rotavirus, 2 kinds of norovirus, intestinal adenovirus, astrovirus and sapovirus. ResultsA total of 1 861 cases of newly diagnosed diarrheal disease were reported, with the peak in July to August. Additionally, 704 surveillance samples were detected, with a total positive detection rate of 50.57%. The detection rates of bacterial, viral and mixed infection were 25.14%, 21.02% and 4.40%, respectively. Among the pathogens detected, DEC accounted for the highest (17.61%, 124/704), followed by norovirus (16.48%, 116/704), rotavirus (6.39%, 45/704), SAL (5.97%, 42/704) and Campylobacter (3.84%, 27/704). DEC detected were mainly enteroaggregative Escherichia coli and enterotoxigenic Escherichia coli, with no detection of Vibrio cholerae, Shigella and YE. The highest total pathogen detection rate was observed from June to September, and the detection peaks of norovirus were from March to June and from October to December, whereas that of DEC was from June to October. The detection rate of rotavirus peaked from January to February, but which was not detected between 2020‒2023. The SAL positive rate peak was in September, whereas that of Campylobacter was from July to September. ConclusionThe main pathogens detected in Fengxian District from 2013‒2019 are DEC, norovirus, rotavirus, SAL and Campylobacter. Different pathogens have different detection peaks, with bacteria predominating in summer and viruses in winter and spring. Prevention and control measures should be carried out according to the epidemiological characteristics of different seasons.

Objective:To develop a treatment-related quality of life scale for Chinese patients with multiple myeloma (MM) and to test its reliability and validity.Methods:The initial scale was constructed through a literature search, Delphi expert correspondence, and cognitive testing. This study conducted a preliminary survey of 379 patients with MM and a formal survey of 865 patients from the hematology departments of 155 hospitals nationwide from February 2024 to March 2024. The final scale was obtained after conducting item analysis and reliability and validity tests on the initial scale.Results:The constructed scale contains 36 items covering six domains: physiological, psychological, social, treatment side effects, general health, and others. In the preliminary survey, the Cronbach’s alpha coefficient of each item ranged from 0.597 to 0.939, and the test-retest reliability was 0.747 ( P<0.001). Exploratory factor analysis extracted eight common factors with a cumulative variance contribution of 60.058%. In the formal survey, the Cronbach’s alpha coefficient of each item ranged from 0.484 to 0.930, and the test-retest reliability was 0.835 ( P<0.001). Confirmatory factor analysis revealed a comparative fit index of 0.750, a root-mean-square error of approximation of 0.090, and a root-mean-square residual of 0.067. Conclusion:The treatment-related quality of life scale for Chinese patients with MM designed in this study exhibited good reliability and validity, reflecting the impact of treatment on the quality of life of patients. This scale can provide a reference to clinicians for assessing the disease status of patients.

Objective:To investigate influences of tetrandrine(Tet)on proliferation,apoptosis and immune escape of melanoma cells by regulating cyclic GMP-AMP synthase(cGAS)-stimulator of interferon gene(STING)signal pathway.Methods:MV3 cells were divided into NC group,Tet-L group(5 μmol/L),Tet-M group(10 μmol/L),Tet-H group(15 μmol/L),RU.521(cGAS inhibi-tor)group(1 μmol/L),Tet-H+RU.521 group(15 μmol/L+1 μmol/L).Proliferation of MV3 cells was detected by CCK-8 and plate cloning test;apoptosis of MV3 cells was detected by flow cytometry;Western blot was used to detect expressions of proliferating cell nuclear antigen(PCNA),Bcl-2-associated X protein(Bax),TGF-β,cGAS,STING proteins in MV3 cells.Cells in above groups were co-cultured with NK cells for 48 h through Transwell chamber,and named NC co-culture group,Tet-L co-culture group,Tet-M co-culture group,Tet-H co-culture group,RU.521 co-culture group,Tet-H+RU.521 co-culture group,respectively.ELISA was used to detect levels of IFN-γ and Granzyme B in supernatant of co-cultured cells;change of NK cell killing activity was detected.Results:Compared with NC group,A450,cloning rate,PCNA and TGF-β protein expressions of MV3 cells in Tet-L group,Tet-M group and Tet-H group were decreased,apoptosis rate,Bax,cGAS,STING protein expressions were increased,which was in a dose dependent man-ner,change trend of corresponding indexes in RU.521 group was contrary to the above(P<0.05);compared with NC co-culture group,levels of IFN-γ,Granzyme B and NK cell killing activity in supernatant of Tet-L co-culture group,Tet-M co-culture group and Tet-H co-culture group were increased,which was in a dose-dependent manner,change trend of corresponding indexes in RU.521 co-culture group was opposite to the above(P<0.05);RU.521 attenuated inhibition of high-dose Tet on MV3 cell proliferation,immune escape and its promotion on apoptosis.Conclusion:Tet may inhibit melanoma cell proliferation,immune escape and promote cell apoptosis by activating cGAS-STING signal pathway.

Objective:To elucidate the correlation between phase angle and malnutrition and to determine the cut-off value for phase angle to predict malnutrition in pancreatic cancer patients.Methods:This prospective cross-sectional study enrolled patients with pancreatic cancer hospitalized in the Department of Hepato-pancreato biliary Surgery at Beijing Hospital between December 2021 and March 2024. Baseline data, diet survey results, and body composition of these patients were recorded. Phase angle was measured with the InBody 720(Biospace, Korea). The Global Leadership Initiative on Malnutrition (GLIM) criteria were used to diagnose malnutrition.Results:A total of 110 cases (75 males and 35 females) aged 29-87(63.00±12.37) years were included. The prevalence of malnutrition was 63.6% (70/111) among the patients. Patients were divided into malnutrition ( n=77) and non-malnutrition ( n=33) groups as per the GLIM criteria. Phase angle was significantly lower in the malnutrition group than in the non-malnutrition group ( t=-3.808, P<0.001). Positive correlations were found between phase angle and body mass index, appendicular skeletal muscle mass index, fat-free mass index , fat-free mass, total energy intake, total protein, albumin, and prealbumin (all P<0.05).After adjusting for sex and age, low phase Angle remained an independent risk factor for malnutrition ( OR=3.809, 95% CI: 1.150-12.612, P=0.029). The cut-off values of phase angle for predicting malnutrition were 4.43 for males (with a sensitivity of 0.955, specificity of 0.585, area under the curve of 0.636; 95% CI: 0.508-0.763, P=0.037) and 4.54 for females (with a sensitivity of 1.000, specificity of 0.708, area under the curve of 0.799; 95% CI: 0.653-0.946, P<0.001). Conclusion:Phase angle may serve as a valid and simple indicator of malnutrition in pancreatic cancer patients.

Objective To explore the effects and therapeutic mechanism of electroacupuncture on the levels of polarization markers and inflammatory factors interleukin(IL)-6,IL-4,tumor necrosis factor-alpha(TNF-α),and IL-10 in rats with para-chlorophenylalanine-induced insomnia(PCPA).Methods Fifty healthy specific-pathogen free grade Sprague-Dawley rats,half male and half female,were randomly divided into a blank group(n=10)and a model reserve group(n=40),in which insomnia was induced by intraperitoneal injection of a 500 mg/kg PCPA suspension.Using the random number table method,the 30 successfully modeled rats were divided into three treatment groups of 10 rats/group:model,electroacupuncture,and estazolam.The estazolam group was given estazolam 0.2 mg/(kg·d)by gavage;the electroacupuncture group was given once-daily electroacupuncture at the"Shenmen"and"Sanyinjiao"acupoints,and stimulation at the"Baihui"and"Benshen"acupoints,20 minutes each time,for 7 consecutive days.Following treatment,serum and hypothalamic levels of TNF-α,IL-6,IL-4,and IL-10 were detected using ELISA and Western blot,while immunofluorescence staining was used to detect the presence of Iba-1 in hypothalamic microglia and the co-expression of CD86 and CD163,which are markers for the M1 and M2 subtypes of microglial cells,respectively.Results Compared with the blank group,the model group exhibited prolonged sleep latency(SL)(P＜0.01),shortened sleep duration(ST)(P＜0.05),significantly higher serum and hypothalamic protein levels of IL-6 and TNF-α(P＜0.01),and significantly lower levels of IL-4 and IL-10(P＜0.01).Compared with the model group,the electroacupuncture and estazolam groups exhibited significantly shorter SL(P＜0.01),prolonged ST(P＜0.01),significantly lower serum and hypothalamic protein levels of IL-6 and TNF-α(P＜0.01),and significantly higher IL-4 and IL-10 levels(P＜0.01).IL-6 content was lower in the electroacupuncture group than in the estazolam group(P＜0.05).Compared with the blank group,the model group exhibited significantly enhanced Iba-1/CD86(M1 type)co-expression(P＜0.01)alongside significantly weakened Iba-1/CD163(M2 type)co-expression(P＜0.01).Under electroacupuncture or estazolam intervention,Iba-1/CD86 co-expression was significantly weakened(P＜0.01),and Iba-1/CD163 co-expression was significantly enhanced in the model group(P＜0.05).Conclusions Electroacupuncture effectively improved sleep disturbances in rats,with an underlying mechanism that may involve regulation of microglial polarization,downregulation of pro-inflammatory cytokine levels,upregulation of anti-inflammatory cytokine levels,and alleviation of neuroinflammation,thereby ameliorating sleep.

Forty adult male C57BL/6 mice were randomly divided into four groups:the control group,Salmonella typhimurium(S.typhimurium)(ST)group,phlorizin(PHZ)+S.typhimuri-um(ST)group,and PHZ(80 mg/kg)group,with 10 mice in each group.Morphological observa-tion,HE staining,ELISA,immunofluorescence and Western blot were performed,the results showed that PHZ significantly increased the cecal index,decreased the spleen index of S.typhi-murium-induced mice(P＜0.05),and reduce the pathological damage of cecum in mice.Mean-while,PHZ treatment also significantly reduced colonization of S.typhimurium in the cecum,spleen,mesenteric lymph nodes and liver(P＜0.05).The results of ELISA showed that PHZ treatment also significantly inhibited the S.ty phimurium-induced increase in the expression of IL-1β,INF-γ,TNF-a and IL-6 in the cecum of mice(P＜0.05).Immunofluorescence and Western blot results showed that PHZ significantly increased the protein expression levels of Occludin,Claudin-3,and ZO-1 in the cecal barrier of mice induced by S.typhimurium(P＜0.05).These results con-firmed that phlorizin could improve cecal inflammation and intestinal barrier damage induced by S.typhimurium in mice.