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 By organizing and implementing a laboratory proficiency validation plan for air change rate testing, this study aims to explore proficiency testing approaches in laboratory animal facilities, assess the current status of relevant laboratories regarding standard application and test capabilities, standardize air change rate testing methods, and ensure the accuracy and reliability of test results. Methods From September to November 2023, the National Institutes for Food and Drug Control (NIFDC) organized a laboratory proficiency validation plan for air change rate testing in laboratory animal facilities (Plan Number: NIFDC-PT-417). The proficiency testing was conducted on-site and consisted of two parts: a written test and practical operation. The written test was open-book. True/false questions focused on participants' understanding of specific clauses in relevant standards, while application-based questions assessed their ability to handle data processing in simulated testing scenarios. The practical operation was conducted according to the relevant criteria of the China National Accreditation Service for Conformity Assessment (CNAS). Two laboratory animal rooms were prepared as proficiency testing samples using a sample splitting approach. These rooms underwent uniformity and stability testing according to CNAS requirements and were approved. Participating laboratories were required to conduct three tests on each of the two laboratory animal rooms, complete the testing and calculation of air change rate within the specified timeframe, and submit their test result reports and original records. Results A total of 27 laboratories registered and participated in the proficiency testing. All participating laboratories submitted their results within the designated timeframe, and the outcomes of all tested laboratories were rated as satisfactory. Conclusion This proficiency validation program objectively and scientifically evaluates the air change rate testing capabilities of selected domestic laboratories, effectively promoting the overall improvement of testing capabilities in the industry. It provides technical support for regulatory authorities to standardize testing institutions and offers reliable references for the purchase of testing services. Through this activity, it was identified that some laboratories need to further enhance their calibration of instruments and the utilization of calibration results. Future efforts should focus on refining related standards to improve the accuracy and reliability of testing.

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.

Objective Through organizing a proficiency evaluation activity for animal illuminance testing laboratories,to assess the accuracy of testing data from participating laboratories,and to propose suggestions for testing methods.Methods In September 2024,China National Institute for Food and Drug Control(NIFDC)organised reference laboratories to conduct on-site testing through open registration.A total of 35 laboratories participated,completed on-site testing,and submitted their reports.The samples to be tested were two adjacent guinea pig breeding rooms in the NIFDC laboratory animal facility.In order to reduce the impact of room voltage fluctuations on animal illumination,chip on board(COB)light sources powered by regulated power supply were used,and brightness was adjusted by regulating voltage output.Split-level test sample pairs were used to set animal illuminance in both rooms,and the reference laboratories conducted testing according to Laboratory animals——Environment and housing facilities(GB 14925-2023).Each reference laboratory was assigned a 3-digit random code,conducted 2 tests for each breeding room,and submitted an on-site test report,original records,and illuminance meter verification or calibration certificate.WPS Spreadsheet 12.1 was used to summarize data,Grubbs' test(α=0.01)was used to check the data and remove outliers,and SPSS 26.0 software was used for data processing and graph drawing.The median value from each reference laboratory was used as the specified value,the robust standard deviation was used as the uncertainty,the robust z-score method(both intra-laboratory and inter-laboratory z-scores)was used to evaluate the test results from each laboratory,and the factors affecting results were analyzed from aspects of personnel,equipment,and operational environment.Results Under the detection level of α=0.01,no outliers were identified.The specified values for Room 1 and Room 2 were 17.80 lx and 19.00 Ix,respectively.The results obtained from 34 laboratories were evaluated as"satisfactory",while the results from one laboratory were"unsatisfactory".Through analysis of illuminance meter calibration certificates from various laboratories,it was found that some reference laboratories had insufficient calibration points and coverage,which caused test results to fail to accurately reflect the actual illuminance of the facility.Conclusion The present study reveals that factors such as personnel,equipment,and operation environment have different impacts on animal illuminance measurements.The present study concentrates exclusively on animal illuminance in guinea pig breeding rooms,where the testing environment is relatively dim.Future research should focus on the development of more efficient methods for selecting measurement sites,with the combination of illuminance meters equipped with wireless probes and storage functions to enhance testing efficiency.

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.

Sj?gren's syndrome(SS)is one of the common rheumatic diseases in clinical practice.Modern medicine commonly uses drugs such as artificial tears,saliva,glucocorticoids,immunosuppressants,and biologics to control the condition.Clinical practice has shown that in addition to modern medical basic treatment,the use of traditional Chinese medicine(TCM)can help improve the clinical efficacy of SS.According to the symptoms and signs of Sj?gren's syndrome in TCM,it is classified as"dryness and obstruction",and the core pathogenesis of the disease is spleen deficiency and deficiency of body fluids.Subsequently,toxic and pathogenic factors gather,leading to the decline of internal organs.The initial causes are spleen damage,unstable barrier,and invasion of pathogenic factors.The core link is spleen dysfunction,insufficient body fluid,and dryness arising from it.Spleen deficiency generates evil,obstruction of qi,and lack of body fluids are the root causes of illness.The main treatment method is the"spleen strengthening method",which treats spleen deficiency,dampness and stagnation,and the body fluid is not distributed.The treatment focuses on strengthening the spleen and qi,supplementing the lungs and generating fluids.Spleen deficiency leads to loss of vitality,blood stasis obstructs blood vessels,and the treatment is to strengthen the spleen,soothe the liver,remove blood stasis,and unblock the orifices.The spleen yang is not vigorous,and qi transformation is impaired.The treatment is to invigorate the spleen and warm the stomach,promote yang circulation,and promote diuresis.

Objective To observe the value of ultrasound radiomics based on convolutional neural network(CNN)for predicting effect of neoadjuvant chemotherapy(NAC)for breast cancer.Methods Totally 164 women with breast cancer were retrospectively enrolled and divided into effective group(n=68)and ineffective group(n=96)according to the treatment response,also randomly divided into training set(n=131)and validation set(n=33)at the ratio of 8∶2.Based on ultrasound before NAC,radiomics features of breast cancer were extracted and screened with CNN,radiomics models were constructed with logistic regression(LR),support vector machine(SVM),K-nearest neighbor(KNN),random forest(RF)and multilayer perceptron(MLP),respectively.The best radiomics model was selected,deep learning score(DL-Score)was calculated,and the nomogram was drawn combined with clinical features.Results Among 5 radiomics models,MLP model had the best comprehensive efficacy for predicting effect of NAC for breast cancer,and its sensitivity,specificity and area under the curve(AUC)in training set was 77.78％,92.21％and 0.929,respectively,which in validation set was 78.57％,84.21％and 0.921,respectively.The estrogen or progesterone receptor,human epidermal growth factor receptor 2 and DL-Score were all independent predictors of NAC effect for breast cancer(all P＜0.05).The sensitivity,specificity and AUC of nomogram drawn based on the above independent predictors was 83.30％,92.21％and 0.953 in training set,85.71％,94.74％and 0.955 in validation set,respectively.AUC of the nomogram was slightly higher than that of MLP model,but no significant difference was found(both P＞0.05).The integrated discrimination improvement index showed that adding clinical features(i.e.the above-mentioned immunohistochemically indicators)could improve the predictive performance of radiomics models(P＜0.001).Conclusion Ultrasound radiomics based on CNN could be used to predict effect of NAC for breast cancer.Combining with immunohistochemically indicators might improve their efficacy.

Objective Based on the needs of supportive care,a supportive care program for patients treated with lymphoma chimeric antigen receptor T cell(CAR-T)immunotherapy was constructed.Methods A questionnaire for supportive care program was initially constructed through a literature review and semi-structured interviews,and then refined through two rounds of Delphi method.Results The response rates for two rounds of questionnaires were both 100％.The expert authority coefficients were 0.89 and 0.90 respectively,and the Kendall's coefficient of concordance was 0.253 and 0.262 respectively.The final program included five primary categories(health information support,physiological symptoms and daily living support,psychological support,care and emotional support,and safety support),20 secondary categories,and 50 tertiary categories.Conclusion The supportive care program developed in this study for patients receiving CAR-T immunotherapy for lymphoma has certain scientific and feasibility.

Objective Through organizing a proficiency evaluation activity for animal illuminance testing laboratories,to assess the accuracy of testing data from participating laboratories,and to propose suggestions for testing methods.Methods In September 2024,China National Institute for Food and Drug Control(NIFDC)organised reference laboratories to conduct on-site testing through open registration.A total of 35 laboratories participated,completed on-site testing,and submitted their reports.The samples to be tested were two adjacent guinea pig breeding rooms in the NIFDC laboratory animal facility.In order to reduce the impact of room voltage fluctuations on animal illumination,chip on board(COB)light sources powered by regulated power supply were used,and brightness was adjusted by regulating voltage output.Split-level test sample pairs were used to set animal illuminance in both rooms,and the reference laboratories conducted testing according to Laboratory animals——Environment and housing facilities(GB 14925-2023).Each reference laboratory was assigned a 3-digit random code,conducted 2 tests for each breeding room,and submitted an on-site test report,original records,and illuminance meter verification or calibration certificate.WPS Spreadsheet 12.1 was used to summarize data,Grubbs' test(α=0.01)was used to check the data and remove outliers,and SPSS 26.0 software was used for data processing and graph drawing.The median value from each reference laboratory was used as the specified value,the robust standard deviation was used as the uncertainty,the robust z-score method(both intra-laboratory and inter-laboratory z-scores)was used to evaluate the test results from each laboratory,and the factors affecting results were analyzed from aspects of personnel,equipment,and operational environment.Results Under the detection level of α=0.01,no outliers were identified.The specified values for Room 1 and Room 2 were 17.80 lx and 19.00 Ix,respectively.The results obtained from 34 laboratories were evaluated as"satisfactory",while the results from one laboratory were"unsatisfactory".Through analysis of illuminance meter calibration certificates from various laboratories,it was found that some reference laboratories had insufficient calibration points and coverage,which caused test results to fail to accurately reflect the actual illuminance of the facility.Conclusion The present study reveals that factors such as personnel,equipment,and operation environment have different impacts on animal illuminance measurements.The present study concentrates exclusively on animal illuminance in guinea pig breeding rooms,where the testing environment is relatively dim.Future research should focus on the development of more efficient methods for selecting measurement sites,with the combination of illuminance meters equipped with wireless probes and storage functions to enhance testing efficiency.