Objective: To develop a clinical decision and prescription generation system (CDPGS) specifically for diarrhea in traditional Chinese medicine (TCM), utilizing a specialized large language model (LLM), Qwen-TCM-Dia, to standardize diagnostic processes and prescription generation. Methods: Two primary datasets were constructed: an evaluation benchmark and a fine-tuning dataset consisting of fundamental diarrhea knowledge, medical records, and chain-of-thought (CoT) reasoning datasets. After an initial evaluation of 16 open-source LLMs across inference time, accuracy, and output quality, Qwen2.5 was selected as the base model due to its superior overall performance. We then employed a two-stage low-rank adaptation (LoRA) fine-tuning strategy, integrating continued pre-training on domain-specific knowledge with instruction fine-tuning using CoT-enriched medical records. This approach was designed to embed the clinical logic (symptoms → pathogenesis → therapeutic principles → prescriptions) into the model’s reasoning capabilities. The resulting fine-tuned model, specialized for TCM diarrhea, was designated as Qwen-TCM-Dia. Model performance was evaluated for disease diagnosis and syndrome type differentiation using accuracy, precision, recall, and F1-score. Furthermore, the quality of the generated prescriptions was compared with that of established open-source TCM LLMs. Results: Qwen-TCM-Dia achieved peak performance compared to both the base Qwen2.5 model and five other open-source TCM LLMs. It achieved 97.05% accuracy and 91.48% F1-score in disease diagnosis, and 74.54% accuracy and 74.21% F1-score in syndrome type differentiation. Compared with existing open-source TCM LLMs (BianCang, HuangDi, LingDan, TCMLLM-PR, and ZhongJing), Qwen-TCM-Dia exhibited higher fidelity in reconstructing the “symptoms → pathogenesis → therapeutic principles → prescriptions” logic chain. It provided complete prescriptions, whereas other models often omitted dosages or generated mismatched prescriptions. Conclusion By integrating continued pre-training, CoT reasoning, and a two-stage fine-tuning strategy, this study establishes a CDPGS for diarrhea in TCM. The results demonstrate the synergistic effect of strengthening domain representation through pre-training and activating logical reasoning via CoT. This research not only provides critical technical support for the standardized diagnosis and treatment of diarrhea but also offers a scalable paradigm for the digital inheritance of expert TCM experience and the intelligent transformation of TCM.

Antibiotic adjuvants offer a promising strategy for restoring antibiotic sensitivity, expanding antibacterial spectra, and reducing required dosages. Previously, compound 15 was identified as a potential adjuvant for Polymyxin B (PB) against multidrug-resistant (MDR) Pseudomonas aeruginosa DK2; however, its clinical utility was hindered by high cytotoxicity, uncertain in vivo efficacy, and an unclear synergetic mechanism. To address these challenges, we synthesized and evaluated a series of novel benzamide derivatives, with A22 emerging as a particularly promising candidate. A22 demonstrated potent synergistic activity to PB, minimal cytotoxicity, improved water solubility, and broad-spectrum synergism of polymyxins against various clinically isolated MDR Gram-negative strains. In vivo studies using Caenorhabditis elegans and mouse models further confirmed the efficacy of A22. Moreover, A22 effectively suppressed the development of PB resistance in Pseudomonas aeruginosa DK2. Mechanistic investigations revealed that A22 enhances polymyxins activity by inducing reactive oxygen species production, reducing ATP levels, increasing NOX activity, and inhibiting biofilm formation, leading to bacterial death. These findings position A22 as a highly promising candidate for the development of polymyxin adjuvants, offering a robust approach to combating MDR Gram-negative bacterial infections.

Objective:To explore the formulation patterns and potential targets of traditional Chinese medicine (TCM) in the treatment of pneumoconiosis using data mining, network pharmacology, and Mendelian randomization (MR) .Methods:In November 2023, prescription data for TCM compound treatments of pneumoconiosis from the past five years were retrieved from the China National Knowledge Infrastructure (CNKI), Wanfang, and VIP databases. The formulation patterns of TCM for pneumoconiosis were analyzed based on drug frequency, association rules, and cluster analysis, followed by the identification of core drugs. Network pharmacology and MR analysis were then applied to determine the potential targets for pneumoconiosis treatment.Results:A total of 100 TCM prescriptions were included, involving 182 herbs, with 15 high-frequency herbs. A total of 24 strong association rules were identified, including 16 herbs. Cluster analysis identified three groups of drug combinations. The core drug combination, "Astragalus Root-Codonopsis Root-Tangkuei-Chinese Magnolia Vine Fruit-Danshen Root-Prepared Rehmannia Root" was determined, which interacted with 47 common targets associated with pneumoconiosis. Further protein-protein interaction (PPI) analysis identified 18 core targets, and MR analysis confirmed that Caspase-3 (CASP3) and Cathepsin B (CTSB) were risk factors for pneumoconiosis ( OR=1.35, 1.44; P=0.038, 0.038) . Conclusion:The core drug combination for the treatment of pneumoconiosis with TCM is "Astragalus Root-Codonopsis Root-Tangkuei-Chinese Magnolia Vine Fruit-Danshen Root-Prepared Rehmannia Root". The medication mainly focuses on tonifying lung qi, moistening dryness and stopping coughing, circulating blood and transforming stasis, which are supplemented with herbs for nourishing yin and moistening dryness, warming yang and dissipating cold. Finally, the key targets for treating pneumoconiosis are screened out, providing an important basis for the development of therapeutic drugs for it.

Objective:To explore the formulation patterns and potential targets of traditional Chinese medicine (TCM) in the treatment of pneumoconiosis using data mining, network pharmacology, and Mendelian randomization (MR) .Methods:In November 2023, prescription data for TCM compound treatments of pneumoconiosis from the past five years were retrieved from the China National Knowledge Infrastructure (CNKI), Wanfang, and VIP databases. The formulation patterns of TCM for pneumoconiosis were analyzed based on drug frequency, association rules, and cluster analysis, followed by the identification of core drugs. Network pharmacology and MR analysis were then applied to determine the potential targets for pneumoconiosis treatment.Results:A total of 100 TCM prescriptions were included, involving 182 herbs, with 15 high-frequency herbs. A total of 24 strong association rules were identified, including 16 herbs. Cluster analysis identified three groups of drug combinations. The core drug combination, "Astragalus Root-Codonopsis Root-Tangkuei-Chinese Magnolia Vine Fruit-Danshen Root-Prepared Rehmannia Root" was determined, which interacted with 47 common targets associated with pneumoconiosis. Further protein-protein interaction (PPI) analysis identified 18 core targets, and MR analysis confirmed that Caspase-3 (CASP3) and Cathepsin B (CTSB) were risk factors for pneumoconiosis ( OR=1.35, 1.44; P=0.038, 0.038) . Conclusion:The core drug combination for the treatment of pneumoconiosis with TCM is "Astragalus Root-Codonopsis Root-Tangkuei-Chinese Magnolia Vine Fruit-Danshen Root-Prepared Rehmannia Root". The medication mainly focuses on tonifying lung qi, moistening dryness and stopping coughing, circulating blood and transforming stasis, which are supplemented with herbs for nourishing yin and moistening dryness, warming yang and dissipating cold. Finally, the key targets for treating pneumoconiosis are screened out, providing an important basis for the development of therapeutic drugs for it.

Objective To prepare mouse monoclonal antibodies against the ectodomain of E2 (E2ecto) glycoprotein of Western equine encephalitis virus (WEEV). Methods A prokaryotic expression plasmid pET-28a-WEEV E2ecto was constructed and transformed into BL21 (DE3) competent cells. E2ecto protein was expressed by IPTG induction and presented mainly as inclusion bodies. Then the purified E2ecto protein was prepared by denaturation, renaturation and ultrafiltration. BALB/c mice were immunized with the formulated E2ecto protein using QuickAntibody-Mouse5W as an adjuvant via intramuscular route, boosted once at an interval of 21 days. At 35 days post-immunization, mice with antibody titer above 1×10⁴ were inoculated with E2ecto intraperitoneally, and spleen cells were fused with SP2/0 cells three days later. Hybridoma cells secreting specific monoclonal antibodies were screened by the limited dilution method, and ascites were prepared after intraperitoneal inoculation of hybridoma cells. The subtypes and titers of the antibodies in ascites were assayed by ELISA. The biological activity of the mAb was identified by immunofluorescence assay(IFA) on BHK-21 cells which were transfected with eukaryotic expression plasmid pCAGGS-WEEV-CE3E2E1. The specificity of the antibodies were evaluated with E2ecto proteins from EEEV and VEEV. Results Purified WEEV E2ecto protein was successfully expressed and obtained. Four monoclonal antibodies, 3G6G10, 3D7G2, 3B9E8 and 3D5B7, were prepared, and their subtypes were IgG2c(κ), IgM(κ), IgM(κ) and IgG1(κ), respectively. The titers of ascites antibodies 3G6G10, 3B9E8 and 3D7G2 were 10⁵, and 3D5B7 reached 10⁷. None of the four antibody strains cross-reacted with other encephalitis alphavirus such as VEEV and EEEV. Conclusion Four strains of mouse mAb specifically binding WEEV E2ecto are successfully prepared.
Horses ; Animals ; Mice ; Encephalitis Virus, Western Equine ; Ascites ; Immunosuppressive Agents ; Antibodies, Monoclonal ; Immunoglobulin M

Objective:To explore and analyze differential expressed genes in malignant pleural mesothelioma (MPM) by bioinformatics method, and to study their prognostic value in MPM and their potential role in immunotherapy.Methods:In January 2022, the dataset GSE51024 was downloaded from the GEO database, and MPM (55 cases) and normal tissue (41 cases) samples were obtained. Using R software and HMDD and miRNet database, MPM-related differential genes were screened and co-expressed genes were identified. Co-expressed genes were enriched and functionally annotated, and protein-protein interaction (PPI) networks were constructed and key genes were identified using the STRING database and Cytoscape software. TRRUST and GEPIA databases were used to predict transcription factors of key genes and to analyze prognosis and survival. The correlation between key genes and the degree of infiltration of immune cells was analyzed using TIMER.Results:A total of 435 co-expressed genes were obtained, which were mainly concentrated in the extracellular matrix tissue and the signaling pathways of cell adhesion molecules. Combined with PPI and TRRUST database, seven key MPM prognostic genes were identified. Among them, cyclin 20 (CDC20) , cell cycle checkpoint kinase 1 (CHEK1) , enhancer of Zeste homolog 2 (EZH2) , ribonucleotide reductase subunit M2 (RRM2) , topoisomerase 2A (TOP2A) , ubiquitin like plant homeodomain and ring finger domain 1 (UHRF1) were significantly up-regulated in MPM, while cyclin A1 (CCNA1) was significantly down-regulated. The expressions of CCNA1, CDC20, CHEK1, EZH2, RRM2, TOP2A and UHRF1 genes were significantly associated with MPM overall survival ( P<0.05) . The expressions of CDC20, CHEK1, EZH2, RRM2 and TOP2A genes were positively correlated with B cells and dendritic cells ( P<0.05) , and negatively correlated with neutrophils ( P<0.05) . Conclusion:CCNA1, CDC20, CHEK1, EZH2, RRM2, TOP2A and UHRF1 may be potential prognostic markers in MPM patients, and their expressions may be related to MPM tumor immunity.

Objective:To explore and analyze differential expressed genes in malignant pleural mesothelioma (MPM) by bioinformatics method, and to study their prognostic value in MPM and their potential role in immunotherapy.Methods:In January 2022, the dataset GSE51024 was downloaded from the GEO database, and MPM (55 cases) and normal tissue (41 cases) samples were obtained. Using R software and HMDD and miRNet database, MPM-related differential genes were screened and co-expressed genes were identified. Co-expressed genes were enriched and functionally annotated, and protein-protein interaction (PPI) networks were constructed and key genes were identified using the STRING database and Cytoscape software. TRRUST and GEPIA databases were used to predict transcription factors of key genes and to analyze prognosis and survival. The correlation between key genes and the degree of infiltration of immune cells was analyzed using TIMER.Results:A total of 435 co-expressed genes were obtained, which were mainly concentrated in the extracellular matrix tissue and the signaling pathways of cell adhesion molecules. Combined with PPI and TRRUST database, seven key MPM prognostic genes were identified. Among them, cyclin 20 (CDC20) , cell cycle checkpoint kinase 1 (CHEK1) , enhancer of Zeste homolog 2 (EZH2) , ribonucleotide reductase subunit M2 (RRM2) , topoisomerase 2A (TOP2A) , ubiquitin like plant homeodomain and ring finger domain 1 (UHRF1) were significantly up-regulated in MPM, while cyclin A1 (CCNA1) was significantly down-regulated. The expressions of CCNA1, CDC20, CHEK1, EZH2, RRM2, TOP2A and UHRF1 genes were significantly associated with MPM overall survival ( P<0.05) . The expressions of CDC20, CHEK1, EZH2, RRM2 and TOP2A genes were positively correlated with B cells and dendritic cells ( P<0.05) , and negatively correlated with neutrophils ( P<0.05) . Conclusion:CCNA1, CDC20, CHEK1, EZH2, RRM2, TOP2A and UHRF1 may be potential prognostic markers in MPM patients, and their expressions may be related to MPM tumor immunity.

Objective To establish a method for uranium aerosol sample collection, dry ashing treatment, and laboratory laser-fluorescence measurement in the workplace of uranium processing and fuel fabrication facilities. Methods Through optimization experiments, the effects of sampling flow, sample pH value, and test temperature on uranium aerosol concentration results were studied, and the detection limit, precision, and recovery rate of the method were tested. Results Under the optimal test conditions, the detection limit of the method was 0.025 ng/mL; the minimum detectable concentration of 1 m³ of aerosol samples was 1.25 × 10⁻³ μg/m³; the relative standard deviation (RSD) of the measurement results was less than 5%; the recovery rate was between 96% and 104%. Conclusion The detection limit, precision, and accuracy of the method meet the testing requirements for uranium aerosol samples in the workplace of uranium processing and fuel fabrication facilities.

Objective:To investigate five kinds of tuberculin skin test (TST), tubercle bacillus-antibody(TB-Ab), interferon-γ release assay(IGRA), tubercle bacillus-DNA (TB-DNA) and collection of bacterial centrifugal smears methods, the application value of combined detection in improving the diagnostic efficiency of pneumoconiosis complicated with tuberculosis.Methods:A total of 150 eligible patients with pneumoconiosis from January 2018 to January 2019 in Zhangjiakou Pulmonary Hospital were selected as the research subjects, and all of them underwent TST, TB-Ab, IGRA, TB-DNA and bacterial centrifugal smear detection. Compared the positive rates of five detection methods in pneumoconiosis and its different stages, and compare the proportion of tuberculosis infection and tuberculosis in different stages of pneumoconiosis.Results:Among the 150 patients with pneumoconiosis, 41 cases (27.33%) were with pneumoconiosis complicated with tuberculosis infection, 24 cases (16.00%) with pneumoconiosis complicated with clinically diagnosed pulmonary tuberculosis, 21 cases (14.00%) with pneumoconiosis complicated with confirmed pulmonary tuberculosis, and 45 cases (30.00%) with pneumoconiosis complicated with pulmonary tuberculosis; with the improvement of pneumoconiosis stage, the proportion of pneumoconiosis combined with tuberculosis infection and pulmonary tuberculosis increased significantly ( P<0.05). Compared with TB-Ab, TB-loop-mediated isothermal amplification(LAMP), and interlayered cup collection centrifuge smear method, the overall positive rate of IGRA detection and pneumoconiosis stage Ⅲ were higher ( P<0.05), but there was no significant difference compared with TST detection ( P>0.05). The positive rate of combined detection was higher, but there was no significant difference compared with IGRA detection ( P>0.05). With the increase of pneumoconiosis stage, the positive reaction intensity of TST decreased, and the positive value of TB-Ab and IGRA increased. Conclusions:The combined detection of TST, TB-Ab, IGRA, TB-DNA and bacterial centrifugal smear method can significantly improve the diagnostic efficiency of pneumoconiosis combined with tuberculosis.

Objective:To investigate the effect of abnormal D-dimer level on the prognosis of patients with frontal lobe injury, and to provide a new idea for clinical diagnosis, treatment and prognosis evaluation of frontal lobe injury.Methods:A retrospective analysis was performed on 71 patients with traumatic brain injury (TBI) who were admitted to the Surgical Care Unit of the First Affiliated Hospital of Zhejiang University School of Medicine from November 1, 2020 to February 1, 2021. According to whether TBI involved in the frontal lobe, the patients were divided into the experimental group ( n = 44) and the control group ( n = 27). Clinical data including sex, age, body mass index (BMI), length of hospital stay, ISS trauma score, GCS coma score, qSOFA score, GOS prognostic score and coagulation index test results within 24 h of admission of the two groups were recorded, and the difference of each index between the two groups was compared. Results:① D-dimer levels were significantly higher in the experimental group than in the control group ( P<0.01); ② The incidence of complications in the experimental group was 40.9%, of which the incidence of lower limb vein thrombosis was 18.2%, and the incidence of complications in the control group was 18.5%, of which the incidence of lower limb vein thrombosis was 3.7%; there were statistical differences between the two groups ( P<0.05); ③ The average hospital stay of patients was 10.96 days in the control group, and 15.50 days in the experimental group with a statistically significant difference between the two groups ( P<0.05); ④ The 28-day mortality rate of patients in the experimental group was 18.2% and 7.4% in the control group. There was no significant difference in 28-day mortality between the two groups ( P>0.05); ⑤ The level of D-dimer was negatively correlated with GOS prognostic score in patients with frontal lobe injury ( r=-0.501, P<0.01). Conclusions:Patients with frontal lobe injury have increased D-dimer levels, increased incidence of complications, and longer hospital stay. There is a significant negative correlation between D-dimer level and prognosis in patients with frontal lobe injury. D-dimer can be used as a sensitive indicator to evaluate the prognosis of patients with frontal lobe injury. The higher the D-dimer level, the worse the prognosis of patients.