Objective To explore the adverse event signals of children using macrolide drugs （azithromycin, clarithromycin, and erythromycin）, and provide reference for rational medicine use in clinical practice. Methods Data from children under 12 years old were extracted from the US FAERS database spanning from the first quarter of 2004 to the second quarter of 2023. The adverse drug reaction （ADR） signal mining for three macrolide antibiotics was conducted using the Reporting Odds Ratio （ROR） and Bayesian Confidence Propagation Neural Network （BCPNN） methods. Special emphasis was placed on analyzing and contrasting the differences in adverse events among the three drugs. Results A total of 1 615 reports for children under 12 years old were retrieved from the FAERS database, including 1 024 reports of azithromycin, 460 reports of clarithromycin, and 131 reports of erythromycin. Among azithromycin and erythromycin, there were more reports from boys than girls, while for clarithromycin, there were more reports from girls than boys. Oral administration was the most common route of administration for all three drugs. Regarding the outcome of adverse events reported, azithromycin and clarithromycin were primarily associated with other serious adverse events, whereas erythromycin was mainly associated with hospitalization and other serious adverse events. The number of adverse events reported decreased with increasing age, with a higher number of reports in the 0-3 age group. Using the ROR and BCPNN methods for signal detection, 86 signals were identified for azithromycin, 91 for clarithromycin, and 34 for erythromycin. These signals involved 22 System Organ Classes （SOCs）, with azithromycin mainly concentrated in skin and subcutaneous tissue disorders （n=21）, clarithromycin in gastrointestinal disorders （n=15）, and erythromycin in gastrointestinal disorders （n=8）. Twenty-four signals of moderate to high risk were detected, with 13 for azithromycin, 9 for clarithromycin, and 2 for erythromycin. Conclusion The adverse events induced by the three drugs with different risks in different systems. When clinically treating Mycoplasma pneumoniae pneumonia in children, the risk profiles of drugs in different systems should be considered, and personalized dosing should be implemented.

ObjectiveTo evaluate the antitumor activity of Periplaneta americana extract（PAE） against human-derived lung adenocarcinoma organoids（LUAD-PDOs） and to elucidate its potential mechanism based on transcriptomics. MethodsFresh tumor and adjacent normal tissues from patients with LUAD were collected to construct LUAD-PDOs and normal lung organoid（Nor-PDOs） models using 3D organoid culture technology. The effective intervention concentration of PAE was determined using the cell counting kit-8（CCK-8） assay. Experimental groups included the model group（LUAD-PDOs）， normal group， model administration group（LUAD-PDOs+PAE）， and normal administration group（Nor-PDOs+PAE）. Hematoxylin-eosin（HE） staining was used to observe the pathological structures of PDOs， immunohistochemistry（IHC） was performed to detect the expressions of the proliferation marker Ki-67 and lung adenocarcinoma differentiation markers cytokeratin-7（CK-7） and Napsin A， TUNEL staining was applied to detect cell apoptosis. RNA sequencing（RNA-Seq） was conducted to identify differentially expressed genes（DEGs）， followed by Gene Ontology（GO）， Kyoto Encyclopedia of Genes and Genomes（KEGG）， and Gene Set Enrichment Analysis（GSEA）， alongside protein-protein interaction（PPI） network analysis to screen core mechanisms. Finally， key targets were validated by integrating external database analysis with immunofluorescence（IF）. ResultsNor-PDOs and LUAD-PDOs that highly recapitulated the pathological characteristics of the primary tissues were successfully established. The CCK-8 assay determined that the effective intervention concentration of PAE was 16 g·L^-1. Morphological observation showed that Nor-PDOs exhibited lumen-forming structures， whereas LUAD-PDOs displayed dense， solid structures. CCK-8 and TUNEL assays revealed that， compared with the model group， PAE intervention inhibited the proliferation of LUAD-PDOs and promoted apoptosis in LUAD cells， while showing no significant effect on the viability of Nor-PDOs. Transcriptomic analysis identified 719 DEGs that were significantly reversed after PAE intervention（347 up-regulated and 372 down-regulated）（P<0.05）. GO enrichment analysis indicated that DEGs in the model administration group were significantly enriched in biological processes related to cell cycle regulation compared to the model group. KEGG pathway analysis revealed that PAE affected pathways related to proliferation and metabolism， including pathways in cancer and the p53 signaling pathway. GSEA further confirmed that PAE significantly enhanced the activity of the p53 signaling pathway（P<0.05）. PPI network analysis indicated that breast cancer type 1 susceptibility protein（BRCA1） and checkpoint kinase 1（CHEK1） were the core down-regulated targets in the p53 pathway. IF verified the high expression of BRCA1 and CHEK1 in LUAD-PDOs and their significant downregulation after PAE intervention（P<0.05）. Furthermore， survival analysis based on The Cancer Genome Atlas（TCGA） database indicated that low expression of BRCA1 and CHEK1 was significantly associated with prolonged overall survival in patients with LUAD（P<0.05）. ConclusionPAE effectively inhibits proliferation of LUAD-PDOs and promotes their apoptosis， its anti-tumor mechanism is potentially associated with the activation of the p53 signaling pathway， with BRCA1 and CHEK1 genes likely serving as key downstream targets for the effects of PAE.

OBJECTIVE To investigate the mechanism of Bushen huoxue formula in improving pregnancy outcomes in mice with antiphospholipid antibody-related recurrent spontaneous abortion （aPL-RSA）. METHODS SPF female BALB/c mice were randomly divided into normal control group， adjuvant control group， model group， and Bushen huoxue formula group. The aPL-RSA mouse model was established by immunization with β 2 glycoproteinⅠcombined with Freund’s adjuvant. From gestational day 0， the Bushen huoxue formula group was administered 1.653 6 g/（kg·d） of the prescription by gavage， while the other three groups received an equal volume of normal saline， twice daily for 15 consecutive days. The uterine appearance of pregnant mice was observed； individual embryo weight was recorded， and the embryo resorption rate was calculated. The proportion of activated CD41 + CD62p + platelets in platelet-rich plasma （PRP） was detected. Serum levels of thromboxane B2 （TXB2） and platelet factor 4 （PF4） were measured. Morphological changes of placental tissue were observed. The cell apoptosis rate of placental tissue was detected. The levels of malondialdehyde （MDA） content， tot al superoxide dismutase （T-SOD） activity， the interleukin-1β （IL-1β） and IL-18， the protein expressions of matrix metalloproteinase-3 （MMP-3）， MMP-9， nuclear proliferation antigen-67 （Ki67）， superoxide dismutase 2 （SOD2）， Nod-like receptor protein 3 （NLRP3）， apoptosis-associated speck-like protein containing a CARD （ASC）， and caspase-1， as well as the mRNA expressions of superoxide dismutase 2（SOD2）， NLRP3， ASC and caspase-1 of placental tissue were detected. RESULTS Compared with the normal control group and adjuvant control group， the model group showed significant decreases in individual embryo weight， T-SOD activity of placental tissue， and relative expression levels of SOD2 mRNA and protein （ P ＜0.05）； while significant increases were observed in embryo resorption rate， proportion of activated CD41 + CD62p + platelets in PRP， serum levels of TXB2 and PF4， placental cell apoptosis rate， MDA content in placental tissue， relative protein expressions of MMP-3， MMP-9 and Ki67， relative protein and mRNA expression levels of NLRP3， ASC and caspase-1 as well as IL-1β and IL-18 levels （ P ＜0.05）. The model group also exhibited irregular uterine morphology with localized atrophy in some uterine horns； placental tissue showed fragmentation of decidual cells， extensive vacuolization and necrosis of trophoblast cells in the labyrinthine zone， accompanied by vascular reduction. After intervention with Bushen huoxue formula， the above indicators were significantly reversed （ P ＜0.05）， and uterine morphology and pathological damage were markedly improved. CONCLUSIONS Bushen huoxue formula can effectively improve pregnancy outcomes in aPL-RSA model mice， and its mechanism may be closely related to inhibiting platelet activation， reducing oxidative stress injury， enhancing trophoblast cell function， and suppressing the inflammatory response mediated by the NLRP3 inflammasome pathway.

Objective To put forward relevant national legislative proposals for preventing workplace violence (WPV) in the healthcare industry by comparing the current legal practices of China and the United States. Methods The Workplace Violence Prevention for Health Care and Social Service Workers Act (hereinafter referred to as the "Act") of the United States was translated and analyzed. The relevant normative legal documents in China were systematically reviewed to compare the legislative differences in the prevention and control of WPV against health care workers. Results The Act aims to establish an employer-driven legal framework for WPV prevention and control. China has no specific legislation for WPV, but has established partial legislation for protecting healthcare workers from external violence through various legal practices. The "Act" regards WPV as an occupational hazard and adopts the priority control order to carry out the prevention and control of WPV. In contrast, China's legislation for WPV approach emphasizes public security and undermines occupational health, treating WPV merely as a work-related injury or accident with limited protection. This gap reveals divergent priorities for legal interests. Conclusion China should integrate WPV prevention and control into the occupational health legal framework through revising existing laws, advancing dedicated legislation, and ratifying relevant international conventions, to strengthen the occupational health legal system. All stakeholders should clarify the responsibilities for WPV prevention and control of healthcare workers, and ensure comprehensive legislative response.

[Objective]Using data mining technology to explore the medication rule and clinical experience of QIU Shengliang,Chief TCM physician(referred as Chief QIU),in treating chronic atrophic gastritis(CAG)with liver and stomach Qi stagnation syndrome.[Methods]By collecting data,setting up admission and discharge standards,establishing a database,standardizing data processing,conducting frequency analysis,constructing a Chinese medicine matrix and performing clustering analysis,combined with the clinical experience of Chief QIU,it conducted analysis and induction of his medication rules for CAG of liver and stomach Qi stagnation syndrome.[Results]This study involved 162 prescriptions,171 herbs.The high-frequency drugs were:Magnolia officinalis,Fructus aurantii,ginger pinellia;Among them,the medicinal properties were mainly warm,the medicinal tastes were mainly spicy,bitter and sweet,and the channels were mainly stomach,spleen,lung and liver.The core drug group was pinellia,Magnolia officinalis and Fructus aurantii.Cluster analysis showed that the core ingredients were:ginger pinellia,Magnolia officinalis,Fructus aurantii,dandelion,galangal,knife bean,spiral flower,cuttlebone,water red flower seed,corrugated seed,hedgehog skin,bamboo shavings and wood butterfly.[Conclusion]Chief QIU is skilled in treating CAG liver and stomach Qi stagnation syndrome.He often uses warm-tonifying herbs and prescribes treatments to regulate Qi and promote Yang,with a focus on resolving stagnation,harmonizing the stomach,dispersing phlegm and eliminating lumps.He emphasizes achieving balance between Yin and Yang,promoting the interaction between rising and descending Qi,harmonizing dryness and dampness,and nurturing the body's vital energy.His prescriptions are tailored to the specific method,and the method is based on the patient's symptoms.

[Objective]To summarize the clinical experience of Dr.QIU Shengliang in treating gastro-esophageal reflux disease(GERD)through regulating Qi dynamics,providing reference for traditional Chinese medicine(TCM)syndrome differentiation and treatment of this condition.[Methods]Through clinical apprenticeships and case analysis combined with classical text research,this paper elucidates the core pathogenesis of"Qi movement disorders"in GERD.It systematically presents Dr.QIU's diagnostic framework centering on regulating Qi dynamics,focusing on spleen-stomach-liver coordination while addressing phlegm-stasis complications.The treatment protocol features flexible herbal applications tailored to syndrome patterns,supported with a representative case study.[Results]Guided by the principle of harmonizing the flow of Qi,Dr.QIU's approach integrates three therapeutic strategies:strengthening the spleen and stomach,soothing liver Qi,and resolving phlegm and blood stasis.The subsequent follow-up revealed that the patient's symptoms were effectively relieved,with no recurrence,thereby validating this method.[Conclusion]Dr.QIU's treatment system emphasizes Qi dynamics regulation as the core principle for GERD management,focusing on visceral functional balance through multi-dimensional interventions encompassing herbal therapy,dietary regulation,emotional management and exercise guidance.This holistic approach demonstrates significant clinical efficacy while embodying TCM's characteristic holistic philosophy and syndrome differentiation principles,offering valuable insights for TCM management of GERD.

Breast cancer is one of the most common malignant tumors among women worldwide, with an increasing incidence rate year by year, making it a significant public health concern. With the continuous advancement of tumor biology research, N 6-methyladenosine (m 6A) modification, as an important form of RNA modification, has attracted growing attention. The m 6A modification, the most prevalent RNA modification in eukaryotes, occurs in almost all types of RNA and plays a critical role in the occurrence, progression, and metastasis of breast cancer. It influences cell proliferation, apoptosis, and alterations in the tumor microenvironment, though the specific mechanisms underlying these effects require further in-depth investigation. Moreover, the specific patterns of m 6A modification demonstrate its potential as a novel biomarker for breast cancer, which could provide new directions for early diagnosis and prognosis evaluation.

Objective:To analyze the molecular mechanism of Huangqin Decoction in the treatment of acute lung injury (ALI) with network pharmacology; To conduct experimental validation.Methods:Active compounds and corresponding targets of Huangqin Decoction were retrieved from the TCMIP database. ALI-related targets were obtained from GeneCards, DisGeNet, TTD, and OMIM, and the intersection targets were obtained. The intersection targets were imported into the string database to build the PPI network, and the core targets were obtained through topology analysis by Cytoscape 3.10.1 software. GO and KEGG pathway enrichment analyses were conducted using clusterProfiler software. 16 SD rats were divided into two groups ( n = 8 per group) with random number table method: control and Huangqin Decoction. Rats in the Huangqin Decoction group received Huangqin Decoction by gavage at a dosage of 40 mg/kg, while the control group was administered an equal volume of distilled water. After seven consecutive treatments, drug-containing serum was collected. A549 cells were divided into four groups: control, model, Huangqin Decoction, and received relevant drugs as intervention for 24 h. Levels of SOD, MDA, GSH-Px, IL-1β, TNF-α, and IL-6 in the culture supernatant were measured by ELISA. Apoptosis was analyzed by flow cytometry. The expressions of PI3K, p-PI3K, Akt, p-Akt, mTOR, p-mTOR, LC3Ⅱ/Ⅰ, and Beclin-1 proteins were determined by Western blot. Results:A total of 137 active compounds and 178 common targets were identified in Huangqin Decoction, with TP53, AKT1, STAT3, TNF, IL6, and ESR1 as core nodes. GO enrichment indicated involvement in oxidative stress and responses to lipopolysaccharides, bacterial molecules, and hypoxia. KEGG analysis revealed enrichment in lipid and atherosclerosis, PI3K-Akt signaling pathway, hepatitis, MAPK signaling pathway, prostate cancer, small-cell lung cancer, and mTOR signaling pathway. In cell experiments, compared with the model group, Huangqin Decoction and inhibitor groups showed increased A549xibo ( P<0.05); levels of IL-1β, TNF-α, IL-6, and MDA in the supernatant were reduced ( P<0.05 or P<0.01), while SOD and GSH-Px levels were elevated ( P<0.05 or P<0.01); the apoptosis rate decreased ( P<0.05 or P<0.01); the expressions of LC3-Ⅱ/Ⅰ and Beclin-1 proteins decreased ( P<0.05 or P<0.01), whereas the expressions of p-mTOR/mTOR, p-Akt/Akt, and p-PI3K/PI3K increased ( P<0.05 or P<0.01). Conclusion:Huangqin Decoction exerts protective effects against ALI mainly by reducing cellular autophagy, and its mechanism may be related to the activation of the mTOR/Akt/PI3K signaling pathway.

The identification and optimization of mutations in nanobodies are crucial for enhancing their thera-peutic potential in disease prevention and control.However,this process is often complex and time-consuming,which limit its widespread application in practice.In this study,we developed a work-flow,named Evolutionary-Nanobody(EvoNB),to predict key mutation sites of nanobodies by combining protein language models(PLMs)and molecular dynamic(MD)simulations.By fine-tuning the ESM2 model on a large-scale nanobody dataset,the ability of EvoNB to capture specific sequence features of nanobodies was significantly enhanced.The fine-tuned EvoNB model demonstrated higher predictive accuracy in the conserved framework and highly variable complementarity-determining regions of nanobodies.Additionally,we selected four widely representative nanobody-antigen complexes to verify the predicted effects of mutations.MD simulations analyzed the energy changes caused by these mu-tations to predict their impact on binding affinity to the targets.The results showed that multiple mu-tations screened by EvoNB significantly enhanced the binding affinity between nanobody and its target,further validating the potential of this workflow for designing and optimizing nanobody mutations.Additionally,sequence-based predictions are generally less dependent on structural absence,allowing them to be more easily integrated with tools for structural predictions,such as AlphaFold 3.Through mutation prediction and systematic analysis of key sites,we can quickly predict the most promising variants for experimental validation without relying on traditional evolutionary or selection processes.The EvoNB workflow provides an effective tool for the rapid optimization of nanobodies and facilitates the application of PLMs in the biomedical field.

Objective:To explore the relationship between thyroid hormone sensitivity and metabolic dysfunction-associated steatotic liver disease(MASLD) in a population with normal thyroid function, with a particular focus on sex-specific differences.Methods:This retrospective study included 41 355 euthyroid cases who underwent routine health examinations at the Health Management Centre of the Second Affiliated Hospital of Dalian Medical University from January 2014 to December 2023 were included. The free triiodothyronine(FT 3) to free thyroxine(FT 4) ratio(FT 3/FT 4) was calculated in order to reflect the peripheral sensitivity of the thyroid gland. Similarly, thyroid feedback quantile-based index(TFQI), thyrotrophic thyroxine resistance index(TT 4RI), and the FT 3-based TFQI-derived index(TFQI-FT 3) were calculated in order to reflect the central sensitivity of the thyroid gland. A Logistic regression was employed to analyse the effect of sex-specific thyroid hormone sensitivity indices on the prevalence of MASLD. The restricted cubic spline was used to analyse the non-linear relationship between the thyroid sensitivity hormone indices and MASLD. Furthermore, the correlation between the thyroid hormone sensitivity indices and MASLD in different subgroups was also analysed. Results:The prevalence of MASLD in the study population was 28.8%. After adjusting the model for confounders, the risk of MASLD increased by 7%, 3%, 10%, and 5% for each standard deviation increase in FT 3/FT 4, TFQI, TFQI-FT 3, and TT 4RI in the total population, respectively. The risk of MASLD increased by 6% and 5% for each standard deviation increase in FT 3/FT 4 and TFQI-FT 3 in men, respectively. For each standard deviation increase in FT 3/FT 4, TFQI, TFQI-FT 3, and TT 4RI in women, the risk of MASLD increased by 6%, 5%, 11%, and 5%, respectively. Higher FT 3/FT 4 and TFQI-FT 3 were positively associated with the risk of developing MASLD in men, and higher FT 3/FT 4, TFQI, TFQI-FT 3, and TT 4RI were positively associated with the risk of developing MASLD in women. There was a non-linear, inverted U-shaped relationship between TFQI and risk of MASLD in women. Subgroup analyses showed positive associations between FT 3/FT 4, TFQI, TFQI-FT 3, and MASLD. Conclusions:The thyroid hormone sensitivity indices may provide a basis for clinical prevention and management of MASLD in individuals with normal thyroid function. Additionally, FT 3/FT 4 and TFQI-FT 3 may indicate the risk of MASLD in the general population, while TFQI and TT 4RI are more suitable for assessing the risk of MASLD in women.