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 analyze the expression level of triggering receptor expressed on myeloid cells-1 （TREM-1） in serum and ascites of patients with cirrhotic ascites， and to investigate its correlation with clinical features and inflammatory markers and its role in the diagnosis of infection and prognostic evaluation. MethodsA total of 110 patients with cirrhotic ascites who were hospitalized in The Fifth Hospital of Shijiazhuang from January 2019 to December 2020 were enrolled， and according to the presence or absence of intra-abdominal infection， they were divided into infection group with 72 patients and non-infection group with 38 patients. The patients with infection were further divided into improvement group with 38 patients and non-improvement group with 34 patients. Clinical data and laboratory markers were collected from all patients. Serum and ascites samples were collected， and ELISA was used to measure the level of TREM-1. The independent-samples t test was used for comparison of normally distributed continuous data between two groups； the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups， and the Kruskal-Wallis H test was used for comparison between multiple groups； the chi-square test was used for comparison of categorical data between two groups. A Spearman correlation analysis was used to investigate the correlation between indicators. A multivariate Logistic regression analysis was used to identify the influencing factors for the prognosis of patients with cirrhotic ascites and infection. The receiver operating characteristic （ROC） curve was used to evaluate the diagnostic and prognostic efficacy of each indicator， and the Delong test was used for comparison of the area under the ROC curve （AUC）. ResultsThe level of TREM-1 in ascites was significantly positively correlated with that in serum （r=0.50， P<0.001）. Compared with the improvement group， the non-improvement group had a significantly higher level of TREM-1 in ascites （Z=-2.391， P=0.017） and serum （Z=-2.544， P=0.011）， and compared with the non-infection group， the infection group had a significantly higher level of TREM-1 in ascites （Z=-3.420， P<0.001）， while there was no significant difference in the level of TREM-1 in serum between the two groups （P>0.05）. The level of TREM-1 in serum and ascites were significantly positively correlated with C-reactive protein （CRP）， procalcitonin （PCT）， white blood cell count， and neutrophil-lymphocyte ratio （r=0.288， 0.344， 0.530， 0.510， 0.534， 0.454， 0.330， and 0.404， all P<0.05）. The ROC curve analysis showed that when PCT， CRP， and serum or ascitic TREM-1 were used in combination for the diagnosis of cirrhotic ascites with infection， the AUCs were 0.715 and 0.740， respectively. The multivariate Logistic regression analysis showed that CRP （odds ratio ［OR］=1.019， 95% confidence interval ［CI］： 1.001‍ ‍—‍ ‍1.038， P=0.043） and serum TREM-1 （OR=1.002， 95%CI： 1.000‍ ‍—‍ ‍1.003， P=0.016） were independent risk factors for the prognosis of patients with cirrhotic ascites and infection， and the combination of these two indicators had an AUC of 0.728 in predicting poor prognosis. ConclusionThe level of TREM-1 is closely associated with the severity of infection and prognosis in patients with cirrhotic ascites， and combined measurement of TREM-1 and CRP/PCT can improve the diagnostic accuracy of infection and provide support for prognostic evaluation.

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.

Objective To evaluate the efficacy of an online-to-offline(O2O)health education guided by the'Timing Theory'in improving self-management behaviours among the patients with glaucoma after day surgery.Methods In this randomised controlled study conducted between July and December 2022,70 patients with glaucoma after day surgery in our hospital were assigned to a control group and an experimental group,with 35 patients per group.Patients in control group received routine nursing care,while those in experimental group received O2O health education based on timing theory in addition to the routine nursing care.Outcomes were evaluated using the glaucoma awareness and knowledge questionnaire(GAKQ),self-efficacy to manage chronic disease scale(SEMCD)and glaucoma self-management questionnaire(GSMQ)at baseline,at 1 month and 3 months after surgery.Results A total of 32 patients in the experimental group and 27 in the control group completed the study.The generalised estimating equation(GEE)analysis showed a significant difference,respectively,in total score of GSMQ in interaction effect(F=8.408,P=0.015)and SEMC in time main effect(F=54.660,P<0.001).There were significant differences in total scores of GAKQ in time main effect,inter-group main effect and interaction effect(F=128.483,P<0.001;F=7.991,P<0.05;F=32.652,P<0.001,respectively).At one-month after intervention,the experimental group showed significantly higher GAKQ and SEMCD scores than the control group(Z=-2.004,P<0.05;Z=-2.029,P<0.05,respectively).At 1-and 3-months after intervention,the experimental group demonstrated significantly higher GAKQ scores(Z=-3.987,P<0.001;Z=-4.505,P<0.001,respectively).Conclusion The timing theory based O2O health education significantly improves knowledge of glaucoma,self-efficacy and self-management behaviours among day surgery patients and helps patients better cope with perioperative self-management over day surgery.

Purpose To explore the predictive value of fetal cardiothoracic ratio(CTR)and their Z-scores for homozygous α-thalassemia major in early pregnancy.Materials and Methods Retrospective analysis was conducted on fetuses at high risk of α-thalassemia major(at 11-13?? weeks of gestation)who received prenatal care at the Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region from June 2016 to December 2023.Ultrasound examinations were performed on these fetuses to measure the CTRs[including cardiothoracic diameter ratio(CTDR),cardiothoracic circumference ratio(CTCR)and cardiothoracic area ratio(CTAR)],which were simultaneously converted into Z-scores.The receiver operating characteristic curve was used to evaluate the predictive efficacy of each group of CTRs and their corresponding Z-scores for fetal α-thalassemia major.Results ①A total of 363 high-risk fetuses were included,including 91(25.1%)in the affected group and 272(74.9%)in the control group.②All the CTRs(including CTDR,CTCR and CTAR)and their corresponding Z-scores in the affected group were significantly higher than those in the control group(t=3.705-21.320,all P<0.001).③The area under the curve(AUC)analysis showed that the AUCs of cardiothoracic ratio Z-scores were higher than those of the corresponding cardiothoracic ratios.The AUCs were ranked from largest to smallest as CTDR Z-score,CTDR,CTAR Z-score,CTCR Z-score,CTAR,CTCR,and the CTDR Z-score had the highest predictive efficacy(when the optimal cut-off value was 1.10,AUC=0.959,with a sensitivity of 94.5%and a specificity of 90.4%).Conclusion The predictive efficacy of fetal CTR Z-scores is higher than that of the corresponding CTRs,which are new indicators for effectively predicting homozygous α-thalassemia major in early pregnancy.Among them,the CTDR Z-score has the highest predictive efficacy,which can improve the prenatal diagnosis rate of this disease and reduce some unnecessary invasive procedures.

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 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.

Purpose Discover new prostate cancer-related single nucleotide variants.Methods Tissue wax blocks from 21 prostate cancer patients who underwent radical surgery and had relatively complete clinical data were collected for somatic mutation detection to analyze new mutated genes associated with prostate cancer.The levels of cor-responding proteins in the urine of prostate cancer patients were tested according to the results of the selected genes.Results All 21 prostate cancer patients showed obvious somatic mutations,and the mutation types were dominated by C＞T and G＞A.The number of somatic mutations was 521,of which 27 genes had high mutation proportions(≥2 ca-ses),including ZSWIM6(5/21),FOXA1(4/21),SPTA1(2/21),FAM47C(2/21),FLG2(2/21),PRSS3(2/21),TP53(2/21),FLG(2/21),UBR4(2/21),and the mutations occurring in ZSWIM6 were all deletion muta-tions,and the mutations occurring in FOXA1 were missense mutations,deletion mutations,and deletion insertion mu-tations.The urinary levels of UPF1,SPTA1,and IDH1 proteins of the 10 prostate cancer patients were significantly different than those of the healthy controls.Correlation analysis showed that FOXA1 was positively correlated with UBR4(r=0.669,P=0.001),SPTA1 was positively correlated with FLG2(r=1.000,P＜0.001),FAM47C was positively correlated with PRSS3(r=1.000,P＜0.001),and there was a significant positive correlation between TP53 and FLG(r=1.000,P＜0.001).ZSWIM6 and FOXA1 were not correlated with biochemical recurrence.SP-TA1 mutation affected progression-free survival(PFS)[(66.0±0)months vs(30.0±7.8)months,P=0.008].FAM47C was positively correlated with PFS[(66.0±0)months vs(19.0±0)months,P＜0.001].ZNF676 was correlated with PFS[(66.0±0)months vs(26.0±5.0)months,P=0.008].FLG2 was correlated with PFS[(66.0±0)months vs(30.0±7.8)months,P=0.008].PRSS3 was correlated with PFS[(66.0±0)months vs(19.0±0)months,P＜0.001].Conclusion All 21 prostate cancer patients harbored somatic mutations,including ZSWIM6(5/21)and FOXA1(4/21)mutations.SPTA1,FAM47C,ZNF676,FLG2,and PRSS3 may be associated with prognosis.

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.