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.

Most patients with pancreatic cancer are already in the locally advanced or metastatic stage at initial diagnosis. While systemic chemotherapy provides clinical benefits for those with mid-to-late-stage pancreatic cancer, its efficacy is often limited by patient tolerance. In response to the dual clinical demands of robust antitumor activity and high targeting specificity, antibody-drug conjugate (ADC) has emerged as a promising solution. By conjugating highly selective monoclonal antibodies with potent cytotoxic small-molecule drugs, ADC achieves precise tumor-targeting while minimizing damage to healthy tissues, which thereby improves treatment tolerance. However, due to the complex pathological features of pancreatic cancer, no ADC has yet been approved for clinical use for this disease. A comprehensive evaluation of factors including ADC-specific targets, payload selection, antibody-drug linkage strategies, drug delivery mechanisms, tissue distribution variability, and tumor heterogeneity will be crucial to advancing the clinical translation of ADC for pancreatic cancer treatment.

Objective To explore the compatibility law of amenorrhea and commonly used local herbs in southwest China using hypergraph;To provide drug choice for the clinical treatment of modern TCM amenorrhea and promote the development and utilization of local Chinese materia medica resources.Methods In this study,the data of prescriptions for the treatment of amenorrhea in Southwest China were used as the data source.By constructing formulas related to amenorrhea-hypergraph(AFR-HG),the topological structure of AFR-HG was analyzed,and the core subnet was extracted according to the node weighted hypercentrality.Based on the sub-network AFR-HG-p1 extracted from the top 20 kinds of Chinese materia medica in AFR-HG,the Kumar algorithm was used to divide the efficacy community network.Results A total of 123 amenorrhea prescriptions were included in this study.Angelicae Sinensis Radix,Carthami Flos,Cyperi Rhizoma,Chuanxiong Rhizoma and other drugs with the efficacy of promoting blood circulation and removing blood stasis,soothing liver and regulating qi,and warming and tonifying were the main drugs with high frequency.Three layers of core subnets were extracted layer by layer through weighted hyper-centrality sorting.The first layer contained Angelicae Sinensis Radix,Chuanxiong Rhizoma,Hyperici Sampsonii Herba,etc.;the second layer of drugs were mainly Lycopi Herba,Rubiae Radix et Rhizoma,Bombycis Feculae,etc.;the third layer of drugs were mainly Ginseng Radix et Rhizoma Rubra,Moutan Cortex,Sargentodoxae Caulis and other drugs;based on the top 20 node overdose drugs,community drugs were obtained through community detection:the first community subnet was Armeniacae Semen Amarum,Draconis Sanguis,Corydalis Rhizoma,etc.,the second community subnet was Rubiae Radix et Rhizoma,Rosae Chinensis Flos,Hyperici Sampsonii Herba,etc.,and the third community subnet was Akebiae Caulis,Homalomenae Rhizoma,Bombycis Feculae,etc.Conclusion In prescriptions of amenorrhea in Southwest China,the treatment principles are basically consistent with modern clinical practice.In addition to commonly used drugs,local herbal medicines and similar medicines are also common.It is of great value to explore the rules of prescriptions for the development and utilization of Chinese materia medica resources.

Objective:To systematically review the current registered clinical study schemes in China regarding the combination of TCM and targeted drug therapy for lung cancer; To analyze their strengths and weaknesses; To provide reference for future study.Methods:Chinese Clinical Trial Registry for clinical study schemes combining TCM with targeted drug therapy for lung cancer treatment was retrieved from the inception to July 10, 2024. The general characteristics, study types, intervention measures, and outcome indicators of existing schemes were systematically summarized and analyzed.Results:A total of 15 studies were included, with the earliest study registered in 2013. Registration locations were concentrated in Shanghai, Guangdong, Tianjin, and Zhejiang. Among them, 9 studies received funding from local, national finance, or hospitals; 13 studies passed ethical review, and 12 included informed consent. The design schemes included 10 randomized controlled studies, 1 cohort study, 1 interventional single-arm study, and 3 observational studies. 6 studies had a sample size smaller than 100 cases, and most were single-center trials. Intervention measures primarily involved the combination of Chinese patent medicine or TCM with targeted drugs, with evaluation indicators mainly focusing on clinical symptoms and laboratory indicators. The setting of outcome indicators lacked a unified standard.Conclusions:Since 2013, clinical studies combining TCM with targeted drug therapy for lung cancer have been relatively methodologically sound but face challenges such as small sample sizes and a certain degree of regional concentration, leading to relatively insufficient representativeness. The future direction for improvement lies in multi-center, large-sample, and well-designed clinical trials. It is also necessary to establish a standardized and normalized system for evaluating outcomes. Integrating basic research to clarify the mechanisms of TCM can provide a theoretical basis for the combination of TCM and targeted drugs, which is conducive to enhancing the rigor and scientific nature of clinical trial design and promoting the formation of high-level evidence-based medicine.

Objective To explore the compatibility law of amenorrhea and commonly used local herbs in southwest China using hypergraph;To provide drug choice for the clinical treatment of modern TCM amenorrhea and promote the development and utilization of local Chinese materia medica resources.Methods In this study,the data of prescriptions for the treatment of amenorrhea in Southwest China were used as the data source.By constructing formulas related to amenorrhea-hypergraph(AFR-HG),the topological structure of AFR-HG was analyzed,and the core subnet was extracted according to the node weighted hypercentrality.Based on the sub-network AFR-HG-p1 extracted from the top 20 kinds of Chinese materia medica in AFR-HG,the Kumar algorithm was used to divide the efficacy community network.Results A total of 123 amenorrhea prescriptions were included in this study.Angelicae Sinensis Radix,Carthami Flos,Cyperi Rhizoma,Chuanxiong Rhizoma and other drugs with the efficacy of promoting blood circulation and removing blood stasis,soothing liver and regulating qi,and warming and tonifying were the main drugs with high frequency.Three layers of core subnets were extracted layer by layer through weighted hyper-centrality sorting.The first layer contained Angelicae Sinensis Radix,Chuanxiong Rhizoma,Hyperici Sampsonii Herba,etc.;the second layer of drugs were mainly Lycopi Herba,Rubiae Radix et Rhizoma,Bombycis Feculae,etc.;the third layer of drugs were mainly Ginseng Radix et Rhizoma Rubra,Moutan Cortex,Sargentodoxae Caulis and other drugs;based on the top 20 node overdose drugs,community drugs were obtained through community detection:the first community subnet was Armeniacae Semen Amarum,Draconis Sanguis,Corydalis Rhizoma,etc.,the second community subnet was Rubiae Radix et Rhizoma,Rosae Chinensis Flos,Hyperici Sampsonii Herba,etc.,and the third community subnet was Akebiae Caulis,Homalomenae Rhizoma,Bombycis Feculae,etc.Conclusion In prescriptions of amenorrhea in Southwest China,the treatment principles are basically consistent with modern clinical practice.In addition to commonly used drugs,local herbal medicines and similar medicines are also common.It is of great value to explore the rules of prescriptions for the development and utilization of Chinese materia medica resources.

Objective To objectively classify cancer-related fatigue(CRF)patients with qi and blood deficiency syndrome;To explore possible enrichment characteristics;To provide ideas for precise population identification.Methods A cross-sectional study was conducted,611 CRF patients with qi and blood deficiency syndrome were included from October 2019 to April 2022.Clinical symptoms of patient were documented,including pale or withered-yellow complexion,shortness of breath with reluctance to talk,dizziness,spontaneous sweating,and palpitations.Additionally,blood routine were conducted,including white blood cell(WBC)count,red blood cell(RBC)count,hemoglobin(HGB),platelet(PLT)count,and neutrophil count(NEUT).The degree of fatigue was assessed using the Piper Fatigue Scale(PFS).Data analysis was performed using R 4.2.0.Clustering analysis was performed using the K-mediods method.Results Through unsupervised clustering analysis,the target population was divided into three categories:qi and blood deficiency syndrome leaning towards qi deficiency,qi and blood deficiency syndrome with mild symptom,qi and blood deficiency syndrome leaning towards blood deficiency.In terms of fatigue severity,the first two groups showed comparable levels,while the third group exhibited the most severe fatigue.Regarding objective indicators,the group leaning towards qi deficiency had the lowest WBC count,NEUT count and PLT count,and the group leaning towards blood deficiency had the lowest HGB count and RBC count.The group of qi and blood deficiency syndrome with mild symptom exhibited the highest level of objective indicators among the three.Conclusion By using clustering analysis methods,the study has preliminarily achieved population enrichment and classification based on syndromes,providing ideas for subsequent information enrichment and accurate identification of populations.Concurrently,the anemia index,WBC count and NEUT count demonstrate a inclination towards blood deficiency syndrome and qi deficiency syndrome,respectively.These findings indicate that these indicators could be incorporated into the syndrome diagnostic criteria for CRF to aid in the classification of the CRF population.

Objective To explore the causal association between coagulation function,including von Willebrand factor(vWF),a disintegrin and metalloproteinase with thrombospondin type 1 motif,member 13(ADAMTS13),activated partial thromboplastin time(aPTT),coagulation factor Ⅷ(FⅧ),coagulation factor Ⅺ(FⅪ),coagulation factor Ⅶ(FⅦ),coagulation factor Ⅹ(FⅩ),endogenous thrombin potential(ETP),plasminogen activator inhibitor-1(PAI-1),protein C,and plasmin,and gestational diabetes mellitus(GDM)using two-sample two-way Mendelian randomization(MR),and to provide genetic evidence for the association between coagulation function and the pathogenesis of GDM.Methods The IEU OpenGWAS database was accessed using the R package TwoSampleMR(v 0.5.6)to obtain the statistical data of the genome-wide association study(GWAS)summary of GDM.MR analysis of the causal association between 11 coagulation function and GDM was performed by the inverse-variance weighted method(IVW),the MR-Egger method,and the weighted median method(WM).Results In this study,the GWAS summary statistics of GDM(covering 5687 cases and 117892 controls)were used for MR analysis.It was found that there was a causal relationship between the predicted plasma FⅧ level and the risk for GDM(IVW:[odds ratio,OR]=0.28,95%confidence interval[CI]:0.10-0.75,P＜0.001;WM:OR=0.30,95%CI:0.09-0.98,P＜0.001).There was no causal relationship between other coagulation function and the risk for GDM(P＞0.05).Conclusion There is a significant causal relationship between the plasma FⅧ level and the risk for GDM.This finding highlights the complex interaction between coagulation function and glucose metabolism during pregnancy,but further research on this finding is warranted.

A 55-year-old male patient was treated with tacrolimus (2.5 mg in the morning and 2 mg at night) for 6 months after lung transplantation to prevent rejection. The blood trough concentration of tacrolimus was stable at 8.0-10.0 μg/L. The patient received antiviral treatment due to corona virus disease 2019 (nirmatrelvir/ritonavir 300 mg/100 mg twice daily orally for a total of 5 days). During the antiviral treatment, the patient continued the anti-rejection treatment. On the second day of antiviral treatment, the patient′s blood trough concentration of tacrolimus increased to >40.0 μg/L, which was considered to be caused by the interaction between nirmatrelvir/ritonavir and tacrolimus. Tacrolimus was withdrawn and antiviral therapy was continued. After discontinuation of tacrolimus for 8 days and nirmatrelvir/ritonavir for 3 days, the blood trough concentration of tacrolimus decreased to 25.7 μg/L. After re-giving tacrolimus at reducing dosage for 3 days, the blood trough concentration of tacrolimus was 8.3 μg/L. After 13 days of resuming administration of tacrolimus at the original dose and frequency, the patient′s blood trough concentration of tacrolimus was 9.2 μg/L. Since then, the blood trough concentration of tacrolimus in the patient was not abnormal again.

A 55-year-old male patient was treated with tacrolimus (2.5 mg in the morning and 2 mg at night) for 6 months after lung transplantation to prevent rejection. The blood trough concentration of tacrolimus was stable at 8.0-10.0 μg/L. The patient received antiviral treatment due to corona virus disease 2019 (nirmatrelvir/ritonavir 300 mg/100 mg twice daily orally for a total of 5 days). During the antiviral treatment, the patient continued the anti-rejection treatment. On the second day of antiviral treatment, the patient′s blood trough concentration of tacrolimus increased to >40.0 μg/L, which was considered to be caused by the interaction between nirmatrelvir/ritonavir and tacrolimus. Tacrolimus was withdrawn and antiviral therapy was continued. After discontinuation of tacrolimus for 8 days and nirmatrelvir/ritonavir for 3 days, the blood trough concentration of tacrolimus decreased to 25.7 μg/L. After re-giving tacrolimus at reducing dosage for 3 days, the blood trough concentration of tacrolimus was 8.3 μg/L. After 13 days of resuming administration of tacrolimus at the original dose and frequency, the patient′s blood trough concentration of tacrolimus was 9.2 μg/L. Since then, the blood trough concentration of tacrolimus in the patient was not abnormal again.

Objective:To investigate the relationship between red blood cell distribution width (RDW) and activity of Crohn′s disease (CD), and explore the diagnostic value of RDW for CD activity.Methods:A cross-sectional study was conducted. Clinical data of CD patients treated continuously at the Sixth Affiliated Hospital of Sun Yat-sen University from November 2012 to October 2014 (CD group) were collected, including the first results of routine hematological examinations and C-reactive protein (CRP) levels after admission. Routine hematological examination results were also collected from healthy peoples undergoing regular health check-ups at the same time, who served as the normal control group. CD patients were divided into remission, mild activity, and moderate-severe activity groups based on the Crohn′s disease activity index (CDAI). The levels of routine hematological indicators including RDW and platelet-to-lymphocyte ratio (PLR) were compared between the CD group and the normal control group, as well as among different subgroups of CD patients. The correlations between RDW, PLR and CD activity or CRP were analyzed, and the diagnostic value of RDW for CD activity was evaluated by using ROC curve. Logistic univariate and multivariate regressions were performed to analyze the influencing factors of the activity of CD. Logistic regression equation was constructed to calculate the diagnostic efficacy of the influencing factors.Results:A total of 303 CD patients (216 males, 87 females; mean age 28.6 ± 11.7 years) were assigned to CD group and 293 healthy peoples (190 males, 103 females; mean age 30.1 ± 12.3 years) were assigned to normal control group. There was no significant differences in age and gender between the two groups (both P>0.05), indicating comparability. Among the CD patients, 109 were in remission group, 106 in mild activity group, and 88 in moderate-severe activity group. There was no significant differences in age and gender among the three subgroups (all P>0.05), indicating comparability. Compared with the normal control group, RDW (15.26% ± 2.51% vs. 13.10% ± 1.13%, P<0.001), PLR (245.09 ± 158.69 vs. 119.07 ± 36.52, P<0.001), neutrophil-to-lymphocyte ratio (NLR) [3.22 (2.06, 4.75) vs. 1.76 (1.39, 2.32), P<0.001], white blood cell count [ (7.68 ± 3.30) ×10 9/L vs. (6.52 ± 1.68) × 10 9/L, P<0.001] and platelet count [ (320.69 ± 116.10) × 10 12/L vs. (230.10 ± 51.08) × 10 12/L, P<0.001] were significantly higher in the CD group, while hemoglobin [ (112.8 ± 21.0) g/L vs. (137.1 ± 13.5) g/L, P<0.001] and platelet distribution width (PDW) [ (10.70 ± 1.91) fl vs. (11.89 ± 1.75) fl, P<0.001] were significantly lower. Compared with patients in remission group, the patients in mild activity group had higher RDW and platelet count, and lower hemoglobin (all P<0.05). Compared with the patients in remission and mild activity groups, the patients in moderate-severe activity group had higher RDW, PLR and NLR, and lower hemoglobin (all P<0.05). Correlation analysis showed the positive correlations between RDW, PLR and CD activity ( r = 0.423, P<0.001; r = 0.295, P<0.001), and RDW was positively correlated with CRP ( r = 0.438, P<0.001). The cut-off value of RDW for predicting CD activity was 13.85%, while the area under curve (AUC) was 0.723 (95% CI: 0.664-0.782, P<0.001), sensitivity was 0.784 and specificity was 0.550. Logistic multivariable regression analysis showed that RDW ( OR = 1.532, 95% CI: 1.291-1.818, P<0.001) and PLR ( OR = 1.003, 95% CI: 1.001-1.006, P = 0.013) were independent risk factors. The combination of RDW and PLR in diagnosing CD activity yielded an AUC of 0.730 (95% CI: 0.673-0.787, P<0.001), sensitivity of 0.407, and specificity of 0.927. Conclusions:CD patients with high RDW have more severe activity. RDW is a simple and practical indicator for diagnosing the activity of CD.