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 develop and validate a risk prediction model for infiltration/extravasation in peripheral intravenous catheter therapy.Methods This retrospective study analyzed 942 patients who completed the Infiltration/Extravasation Risk Assessment Form between January and June 2023 at the First Affiliated Hospital of Army Medical University(including Departments of Neurology,Endocrinology,Gastroenterology and Hepatobiliary Surgery).Patients were allocated to a derivation cohort(n=628)and validation cohort(n=314)in a 2∶1 ratio based on catheterization chronology.The derivation cohort served for model development and internal validation,while the validation cohort underwent external validation.Logistic scoring method constructed the risk model,with Hosmer-Lemeshow(HL)test assessing goodness-of-fit and ROC curve evaluating predictive performance.Twenty-one potential risk factors were assessed,including age,gender,chronic diseases,clinician experience,treatment compliance,and total infusion volume.Results Infiltration/extravasation occurred in 48 cases(5.10%incidence:31 derivation/17 validation).Among 21 factors,15 showed significant association(P<0.05),with 6 independent predictors:junior high school education or below(OR=5.2),chronic disease history(OR=3.1),poor compliance(OR=2.8),lower extremity venipuncture(OR=4.1),total infusion≥1 000 mL(OR=3.5),and hyperosmotic/corrosive medications(OR=6.7).The final prediction model was:Y=2×(low education)+1×(chronic disease)+1×(poor compliance)+1×(lower extremity puncture)+1×(volume≥1 000 mL)+2×(corrosive agents).For the derivation cohort,AUC was 0.967(95%CI:0.936～0.998),specificity 0.911,sensitivity 0.935,with good calibration(χ2=4.135,P=0.845).Validation cohort showed AUC 0.939(0.853～1.000),specificity 0.919,sensitivity 0.941,and acceptable calibration(χ2=8.998,P=0.085).Conclusion This model demonstrates excellent discriminative ability and calibration,providing an effective tool for identifying high-risk patients and guiding targeted preventive strategies.

Objective In this study,2,4-dinitrochlorobenzene(DNCB)was used to induce the establishment of an atopic dermatitis(AD)model in BALB/c homozygous mice to simulate the skin inflammatory complications in patients with clinical malignancies.Methods BALB/c mice were divided into different groups:negative control group(NC group),model group(MODEL group),atopic dermatitis group(AD group),and dexamethasone group(DEX group).After the mice in MODEL group and DEX group were inoculated with S-180 tumor cells in the axilla,MODEL group,AD group and DEX group were stimulated with DNCB on the dorsal skin and the ear to establish an animal model of atopic dermatitis in tumor-bearing mice.Changes in body weight were observed and recorded,the dorsal skin condition of mice was assessed after the last administration of the drug,the spleen was taken to calculate the spleen coefficient,the difference in the mass of mouse ear slices was determined to calculate the degree of auricular swelling and the rate of inhibition of swelling,and histopathological tests were performed on the dorsal skin tissues to detect the levels of IgE,TNF-α,IL-4,and IL-17 in the serum using an ELISA assay.Results Compared with the NC group,the skin of mice in the MODEL and AD groups showed erythematous,papular,scaly and mossy changes,accompanied by weight loss,and a significant increase in splenic coefficient and auricular swelling.Pathologic findings showed an incomplete skin structure,a significant increase in skin thickness,a large infiltration of inflammatory cells,and an increase in the number of mast cells.Serum levels of IgE,TNF-α,IL-4 and IL-17 were increased.Compared with the MODEL group,the DEX group showed an improvement in all the assays.Conclusions DNCB excitation can successfully establish an animal model of AD in hormonal mice,which is drug-controllable,which provides a useful scientific tool for conducting scientific research related to malignant tumors and skin inflammation.

The underlying cause of low response rates to existing immunotherapies is that tumor cells dominate tumor immune escape through surface antigen deficiency and inducing tumor immunosuppressive microenvironment (TIME). Here, we proposed an in situ tumor cell engineering strategy to disrupt tumor immune escape at the root by restoring tumor cell MHC-I/tumor-specific antigen complex (MHC-I/TSA) expression to promote T-cell recognition and by silencing tumor cell CD55 to increase the ICOSL⁺ B-cell proportion and reverse the TIME. A doxorubicin (DOX) and dual-gene plasmid (MAC pDNA, encoding both MHC-I/ASMTNMELM and CD55-shRNA) coloaded drug delivery system (LCPN@ACD) with tumor targeting and charge/size dual-conversion properties was prepared. LCPN@ACD-induced ICD promoted DC maturation and enhanced T-cell activation and infiltration. LCPN@ACD enabled effective expression of MHC-I/TSA on tumor cells, increasing the ability of tumor cell recognition and killing. LCPN@ACD downregulated tumor cell CD55 expression, increased the proportion of ICOSL⁺ B cells and CTLs, and reversed the TIME, thus greatly improving the efficacy of αPD-1 and CAR-T therapies. The application of this in situ tumor cell engineering strategy eliminated the source of tumor immune escape, providing new ideas for solving the challenges of clinical immunotherapy.

Objective:To determine the optimal threshold for trigger-day progesterone levels in gonadotropin-releasing hormone (GnRH) antagonist protocols.Methods:A cohort study was performed. The clinical data were retrospectively analyzed from patients who underwent in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) for assisted reproduction at the Reproductive Medicine Center of Tianjin Central Obstetrics and Gynecology Hospital between January 2014 and May 2023. The study included 5 760 fresh transfer cycles where the female partner had undergone ovarian stimulation using a GnRH antagonist protocol. This was a single-arm study. The primary outcome measures were clinical pregnancy rate and live birth rate. The association between progesterone level on the trigger day and clinical pregnancy outcome and the dose-response relationship were analyzed by restricted cubic spline. Results:The progesterone level on the day of human chorionic gonadotropin trigger was (1.33±0.38) μg/L. Among the included cycles, 2 900 cycles underwent conventional IVF fertilization, while 2 860 cycles underwent ICSI. The biochemical pregnancy rate was 44.79% (2 580/5 760), the clinical pregnancy rate was 40.35% (2 324/5 760), and the live birth rate was 31.46% (1 812/5 760). Progesterone levels on the trigger day in GnRH antagonist protocols showed a nonlinear association with both clinical pregnancy rate and live birth rate (both P<0.001). When progesterone levels were below 0.61 μg/L, the clinical pregnancy rate increased with rising progesterone levels, but decreased significantly once this threshold was exceeded. Similarly, the live birth rate increased with progesterone levels below 0.63 μg/L and declined beyond that point. Conclusion:Progesterone levels on the optimal trigger day for achieving the highest clinical pregnancy rate and live birth rate in GnRH antagonist protocols are peak values of 0.61 μg/L and 0.63 μg/L, respectively. Using these thresholds, the impact of progesterone levels on the trigger day shows a positive effect on both clinical pregnancy rate and live birth rate up to these points, after which the effects become negative.

Objective:To investigate the association between different socioeconomic status (SES) levels and the incidence of sensory impairment (SI) in the Chinese population.Methods:This study adopted a prospective cohort design, utilizing data from the China Health and Retirement Longitudinal Study (CHARLS) collected in July or August 2011. Participants who met the inclusion and exclusion criteria were followed up every 2-3 years until the onset of SI or the end of the follow-up period (August 2018). Based on educational attainment and annual per capita household expenditure, participants were classified into four SES groups: low, lower-middle, upper-middle, and high SES. Logistic regression was employed to analyze the relationship between different SES levels and the incidence of SI.Results:A total of 7 415 participants were included in the study, with a mean follow-up duration of 4.9 years. A total of 3 644 cases of incident SI were recorded (49.1%). Compared with the high SES group, the risk of developing SI was progressively higher in the upper-middle SES group ( OR=1.42, 95% CI: 1.03-1.96), lower-middle SES group ( OR=1.83, 95% CI: 1.29-2.60), and low SES group ( OR=2.04, 95% CI: 1.42-2.94) ( P for trend<0.001). Conclusions:SES is closely associated with new-onset SI. Enhancing SES may help reduce the risk of developing SI.

Objective:To determine the optimal threshold for trigger-day progesterone levels in gonadotropin-releasing hormone (GnRH) antagonist protocols.Methods:A cohort study was performed. The clinical data were retrospectively analyzed from patients who underwent in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) for assisted reproduction at the Reproductive Medicine Center of Tianjin Central Obstetrics and Gynecology Hospital between January 2014 and May 2023. The study included 5 760 fresh transfer cycles where the female partner had undergone ovarian stimulation using a GnRH antagonist protocol. This was a single-arm study. The primary outcome measures were clinical pregnancy rate and live birth rate. The association between progesterone level on the trigger day and clinical pregnancy outcome and the dose-response relationship were analyzed by restricted cubic spline. Results:The progesterone level on the day of human chorionic gonadotropin trigger was (1.33±0.38) μg/L. Among the included cycles, 2 900 cycles underwent conventional IVF fertilization, while 2 860 cycles underwent ICSI. The biochemical pregnancy rate was 44.79% (2 580/5 760), the clinical pregnancy rate was 40.35% (2 324/5 760), and the live birth rate was 31.46% (1 812/5 760). Progesterone levels on the trigger day in GnRH antagonist protocols showed a nonlinear association with both clinical pregnancy rate and live birth rate (both P<0.001). When progesterone levels were below 0.61 μg/L, the clinical pregnancy rate increased with rising progesterone levels, but decreased significantly once this threshold was exceeded. Similarly, the live birth rate increased with progesterone levels below 0.63 μg/L and declined beyond that point. Conclusion:Progesterone levels on the optimal trigger day for achieving the highest clinical pregnancy rate and live birth rate in GnRH antagonist protocols are peak values of 0.61 μg/L and 0.63 μg/L, respectively. Using these thresholds, the impact of progesterone levels on the trigger day shows a positive effect on both clinical pregnancy rate and live birth rate up to these points, after which the effects become negative.

Objective To explore the syndrome differentiation rules of the Yin Dan and Yang Dan method,and summarize Professor Yang Zhimin's valuable experience in the treatment of insomnia.Methods The case data of 11390 insomnia patients treated by Professor Yang Zhimin was retrospectively analyzed,and the distribution of symptoms,signs,pathogenesis and syndrome differentiation rules of insomnia patients was examined based on latent structure analysis.Result A total of 44 symptoms and signs can explain the overall situation of insomnia patients,showing the characteristics of both excess and deficiency,cold and heat.A total of 36 hidden categories(symptom clusters and pathogenesis)was composed.Symptoms and signs such as fear of wind and cold can explain the overall situation of insomnia patients with cold syndrome due to weakness of Wei Qi,symptoms and signs such as pale eyelid can explain the overall situation of insomnia patients with heat syndrome due to weakness of Ying Xve,and established diagnostic criteria for insomnia patients with cold syndrome due to weakness of Wei Qi and heat syndrome due to weakness of Ying Xve.Conclusion Based on the Yang Dan and Yin Dan method to distinguish and treat insomnia from the perspective of disharmony between Ying and Wei,it is in line with the basic understanding of the pathogenesis of insomnia in the classics of traditional Chinese medicine.The pattern differentiation rules obtained through the latent structure analysis method are helpful to summarize valuable clinical experience and provide a certain reference and basis for clinical practice.

Objective To explore the syndrome differentiation rules of the Yin Dan and Yang Dan method,and summarize Professor Yang Zhimin's valuable experience in the treatment of insomnia.Methods The case data of 11390 insomnia patients treated by Professor Yang Zhimin was retrospectively analyzed,and the distribution of symptoms,signs,pathogenesis and syndrome differentiation rules of insomnia patients was examined based on latent structure analysis.Result A total of 44 symptoms and signs can explain the overall situation of insomnia patients,showing the characteristics of both excess and deficiency,cold and heat.A total of 36 hidden categories(symptom clusters and pathogenesis)was composed.Symptoms and signs such as fear of wind and cold can explain the overall situation of insomnia patients with cold syndrome due to weakness of Wei Qi,symptoms and signs such as pale eyelid can explain the overall situation of insomnia patients with heat syndrome due to weakness of Ying Xve,and established diagnostic criteria for insomnia patients with cold syndrome due to weakness of Wei Qi and heat syndrome due to weakness of Ying Xve.Conclusion Based on the Yang Dan and Yin Dan method to distinguish and treat insomnia from the perspective of disharmony between Ying and Wei,it is in line with the basic understanding of the pathogenesis of insomnia in the classics of traditional Chinese medicine.The pattern differentiation rules obtained through the latent structure analysis method are helpful to summarize valuable clinical experience and provide a certain reference and basis for clinical practice.

The incidence of cutaneous squamous cell carcinoma (CSCC) is increasing rapidly. This study discussed the effects of artesunate (ART) on CSCC cell proliferation and migration via the solute carrier family 7 member 11 (SLC7A11)-glutathione peroxidase 4 (GPX4) pathway. MTT assessed cell viability and analyzed the IC50 value (69.26 μM). Accordingly, human CSCC cells (A431) were cultured in vitro, and treated with 70 μM ART, Ferrostatin-1, oe-SLC7A11, and C646, with cell biological behavior assessed.The potential targets of ART were predicted. p53 acetylation and protein stability and ART-p300 binding were examined. Thymusless nude mice were subcutaneously inoculated with A431 cells, and treated with ART and C646. ART-treated A431 cells showed weakened proliferation, migration, lactate dehydrogenase levels, oxidized glutathione/glutathione ratio, reactive oxygen species, malondialdehyde, and active Fe2+ levels, which could be reversed by suppressing ferroptosis. ART promoted p53 acetylation and protein stability and curbed the SLC7A11-GPX4 pathway by targeting p300. ART stimulated ferroptosis via the SLC7A11-GPX4 pathway, thereby repressing CSCC cell proliferation and migration, which were counteracted by p300 inhibition. ART regulated the SLC7A11-GPX4 pathway by up-regulating the p300-p53 axis, thereby hindering tumor growth in vivo. Collectively, ART inhibits CSCC proliferation and migration by modulating the SLC7A11-GPX4 pathway through the p300-p53 axis.