Objective: To develop QingNangTCM, a specialized large language model (LLM) tailored for expert-level traditional Chinese medicine (TCM) question-answering and clinical reasoning, addressing the scarcity of domain-specific corpora and specialized alignment. Methods: We constructed QnTCM_Dataset, a corpus of 100 000 entries, by integrating data from ShenNong_TCM_Dataset and SymMap v2.0, and synthesizing additional samples via retrieval-augmented generation (RAG) and persona-driven generation. The dataset comprehensively covers diagnostic inquiries, prescriptions, and herbal knowledge. Utilizing P-Tuning v2, we fine-tuned the GLM-4-9B-Chat backbone to develop QingNangTCM. A multi-dimensional evaluation framework, assessing accuracy, coverage, consistency, safety, professionalism, and fluency, was established using metrics such as bilingual evaluation understudy (BLEU), recall-oriented understudy for gisting evaluation (ROUGE), metric for evaluation of translation with explicit ordering (METEOR), and LLM-as-a-Judge with expert review. Qualitative analysis was conducted across four simulated clinical scenarios: symptom analysis, disease treatment, herb inquiry, and failure cases. Baseline models included GLM-4-9B-Chat, DeepSeek-V2, HuatuoGPT-II (7B), and GLM-4-9B-Chat (freeze-tuning). Results: QingNangTCM achieved the highest scores in BLEU-1/2/3/4 (0.425/0.298/0.137/0.064), ROUGE-1/2 (0.368/0.157), and METEOR (0.218), demonstrating a balanced and superior normalized performance profile of 0.900 across the dimensions of accuracy, coverage, and consistency. Although its ROUGE-L score (0.299) was lower than that of HuatuoGPT-II (7B) (0.351), it significantly outperformed domain-specific models in expert-validated win rates for professionalism (86%) and safety (73%). Qualitative analysis confirmed that the model strictly adheres to the “symptom-syndrome-pathogenesis-treatment” reasoning chain, though occasional misclassifications and hallucinations persisted when dealing with rare medicinal materials and uncommon syndromes. Conclusion Combining domain-specific corpus construction with parameter-efficient prompt tuning enhances the reasoning behavior and domain adaptation of LLMs for TCM-related tasks. This work provides a technical framework for the digital organization and intelligent utilization of TCM knowledge, with potential value for supporting diagnostic reasoning and medical education.

OBJECTIVE To establish a preparation system for megakaryocytes(MKs)derived from human embryonic stem cells(hESCs)and MK microparticles(MKMPs),and to assess the pro-angio-genic efficiency of these microparticles.METHODS ①hESCs were induced to mesodermal progenitor cells via monolayer culture with the first-stage induction medium for 2 days before the cells were induced to hemogenic endothelial/hematopoietic progenitor cells by culturing with the second-stage induction medium for another 3 days.Then,the cells were dissociated into single cells,seeded into the third-stage induction medium,and cultured using the suspension method for 8 days to obtain MKs.The specific characters of differentiated cells were identified through morphological observation and flow cytometry before stage-specific marker proteins in different periods were analyzed[hESCs:TRA-1-60,sialyl glycolipid stage-specific embryonic antigen4(SSEA4)];mesodermal progenitor cells:brachyury;hemogenic endothelial/hematopoietic progenitor cells:CD34,CD43;MKs:CD41a,CD42b),and immu-nofluorescence staining[β1-tubulin,von Willebrand factor(VWF)],[friend leukemia integration 1(FLI1),CD42].② MKMP collection and verification:MKMPs were collected via differential centrifugation.The concentration and size of these MKMPs were determined by nanoparticle tracking analysis(NTA),and both the morphology and ultrastructure were examined by transmission electron microscopy(TEM).Besides,the MKMPs-specific proteins[CD41,tumor susceptibility gene 101(TSG101)and CD9]were detected by Western blotting analysis.③ Biological function of MKMPs:MKMPs were stained with CD41a-PE antibodies and co-cultured with human umbilical veinvascular endothelial cells(HUVECs)labeled by CD34-APC for 3 h.Live-cell immunofluorescence was employed to find out whether HUVECs could absorb MKMPs.To find out whether MKMPs could affect the role of HUVECs in angio-genesis and cell migration,platelet microvesicles(PMPs)were used as positive controls.The experi-mental groups were added with different concentrations of microparticles(1,5,10 and 20 mg·L-1)while the control group was given no microparticles(0 mg·L-1).The number of nodes that formed the lumen after 5 h of incubation in Matrigel was counted,and the size of healing of the scratch area was analyzed after 6 h.To elucidate the mechanism through which MKMPs impacted angiogenesis,ELISA was used out to quantitatively detect the concentration of proteins in microparticles.RESULTS ① A three-stage differentiation cultural system was established to develop hESCs into MKs.Flow cytometry revealed progressive loss of pluripotency markers SSEA4 and TRA-1-60,while the mesodermal progenitor marker brachyury peaked at d 2.Subsequently,hemogenic endothelial/hematopoietic progenitor markers CD34 and CD43 emerged at d 5,followed by megakaryocytic markers CD41a and CD42b at d 13.Immunofluorescent images further demonstrated that MKs expressed specific proteins CD42,β1-tubulin,von VWF and FLI1 at d 13.②Microparticles were collected via differential centrifuga-tion.Transmission electron microscopy revealed that their substructure exhibited a typical double-layered membrane.Nanoparticle tracking analysis indicated that the size was(164.3±14.0)nm.The result of WB demonstrated that the microparticles expressed specific markers,including TSG101,CD9 and CD41.③ MKMPs were absorbed after being co-cultured with HUVECs for 3 h and enhanced the ability of HUVECs to form tubes and migrate.Notably,the treatment of 5 mg·L-1 MKMPs was more effective than 5 mg·L-1 PMPs treatment.The results of ELISA showed that the content of VEGF from MKMPs was higher than from PMPs,which may be the key factor in regulating endothelial biological function.CONCLUSION MKs derived from hESCs can generate functional microparticles which can promote angiogenesis.

Objective:To develop and evaluate a medical visual question answering (VQA) model for neonatal lung ultrasound (LUS) images to enhance intelligent auxiliary diagnosis of neonatal pulmonary diseases.Methods:Using data from neonates admitted to Beijing Obstetrics and Gynecology Hospital, Capital Medical University (January 2023 to December 2024), an image-question-answer dataset comprising 251 LUS images was constructed [43 pneumonia (17.1%), 42 neonatal respiratory distress syndrome (16.7%), 83 transient tachypnea (33.1%), and 83 normal (33.1%) images] with a four-tier medical question-answer framework. Building upon the Qwen2.5-VL-7B base model and integrating LoRA fine-tuning with chain-of-thought prompting, we developed the NLUS-VQA model to enhance visual-language semantic alignment and enable stepwise clinical reasoning, achieving efficient small-sample adaptation. Model performance was comprehensively assessed through natural language generation metrics (BLEU-4, ROUGE-1/2/L), qualitative evaluation of characteristic recognition, and clinical consistency analysis.Results:(1) Quantitative evaluation demonstrated that NLUS-VQA achieved scores of 22.38 (BLEU-4), 48.26 (ROUGE-1), 22.40 (ROUGE-2), and 37.20 (ROUGE-L), representing significant improvements over baseline models. (2) Qualitatively, the model exhibited strong performance in identifying lung consolidation, coalescent B-lines, and snowflake signs, with its chain-of-thought strategy enhancing clinical interpretability and answer accuracy. (3) Clinically, NLUS-VQA achieved a Cohen's Kappa coefficient of 0.78 and diagnostic accuracy of 80.8% (21/26), indicating substantial agreement with clinical experts.Conclusion:The NLUS-VQA model demonstrates robust interpretability in recognizing key sonographic patterns (e.g. lung consolidation, confluent B-lines, and snowflake signs), providing a scalable framework for small-sample medical image analysis, though diagnostic performance on complex conditions remains limited by dataset scale and minority class representation.

Screening carbonyl reductases with the ability to catalyze the reduction of complex carbonyl compounds is of great significance for the biosynthesis of R-tolvaptan(R-TVP). In this study, the target carbonyl reductase in the crude enzyme extract of rabbit liver was separated, purified, and identified by ammonium sulfate precipitation, gel-filtration chromatography, ion exchange chromatography, affinity chromatography, and protein mass spectrometry. With the rabbit liver genome as the template, the gene encoding the carbonyl reductase rlsr5 was amplified by PCR and the recombinant strain was successfully constructed. After RLSR5 was purified by affinity chromatography, its enzymatic properties were characterized. The results indicated that the gene sequence of rlsr5 was 972 bp, encoding a protein with a molecular weight of 40 kDa. RLSR5 was a dimeric protein, and each monomer was composed of a (α/β)₈-barrel structure. RLSR5 could asymmetrically reduce 7-chloro-1-[2-methyl-4-[(2- methylbenzoyl)amino]benzoyl]-5-oxo-2,3,4,5-tetrahydro-1H-1-benzazepine (prochiral ketone, PK) to synthesize R-TVP. The specific activity of the enzyme was 36.64 U/mg, and the optical purity of the product was 99%. This enzyme showcased the optimal performance at pH 6.0 and 30 °C. It was independent of metal ions, with the activity enhanced by Mn²⁺. This study lays a foundation for the biosynthesis of tolvaptan of optical grade.
Animals ; Rabbits ; Alcohol Oxidoreductases/biosynthesis* ; Recombinant Proteins/metabolism* ; Escherichia coli/metabolism* ; Liver/enzymology*

OBJECTIVE To establish a preparation system for megakaryocytes(MKs)derived from human embryonic stem cells(hESCs)and MK microparticles(MKMPs),and to assess the pro-angio-genic efficiency of these microparticles.METHODS ①hESCs were induced to mesodermal progenitor cells via monolayer culture with the first-stage induction medium for 2 days before the cells were induced to hemogenic endothelial/hematopoietic progenitor cells by culturing with the second-stage induction medium for another 3 days.Then,the cells were dissociated into single cells,seeded into the third-stage induction medium,and cultured using the suspension method for 8 days to obtain MKs.The specific characters of differentiated cells were identified through morphological observation and flow cytometry before stage-specific marker proteins in different periods were analyzed[hESCs:TRA-1-60,sialyl glycolipid stage-specific embryonic antigen4(SSEA4)];mesodermal progenitor cells:brachyury;hemogenic endothelial/hematopoietic progenitor cells:CD34,CD43;MKs:CD41a,CD42b),and immu-nofluorescence staining[β1-tubulin,von Willebrand factor(VWF)],[friend leukemia integration 1(FLI1),CD42].② MKMP collection and verification:MKMPs were collected via differential centrifugation.The concentration and size of these MKMPs were determined by nanoparticle tracking analysis(NTA),and both the morphology and ultrastructure were examined by transmission electron microscopy(TEM).Besides,the MKMPs-specific proteins[CD41,tumor susceptibility gene 101(TSG101)and CD9]were detected by Western blotting analysis.③ Biological function of MKMPs:MKMPs were stained with CD41a-PE antibodies and co-cultured with human umbilical veinvascular endothelial cells(HUVECs)labeled by CD34-APC for 3 h.Live-cell immunofluorescence was employed to find out whether HUVECs could absorb MKMPs.To find out whether MKMPs could affect the role of HUVECs in angio-genesis and cell migration,platelet microvesicles(PMPs)were used as positive controls.The experi-mental groups were added with different concentrations of microparticles(1,5,10 and 20 mg·L-1)while the control group was given no microparticles(0 mg·L-1).The number of nodes that formed the lumen after 5 h of incubation in Matrigel was counted,and the size of healing of the scratch area was analyzed after 6 h.To elucidate the mechanism through which MKMPs impacted angiogenesis,ELISA was used out to quantitatively detect the concentration of proteins in microparticles.RESULTS ① A three-stage differentiation cultural system was established to develop hESCs into MKs.Flow cytometry revealed progressive loss of pluripotency markers SSEA4 and TRA-1-60,while the mesodermal progenitor marker brachyury peaked at d 2.Subsequently,hemogenic endothelial/hematopoietic progenitor markers CD34 and CD43 emerged at d 5,followed by megakaryocytic markers CD41a and CD42b at d 13.Immunofluorescent images further demonstrated that MKs expressed specific proteins CD42,β1-tubulin,von VWF and FLI1 at d 13.②Microparticles were collected via differential centrifuga-tion.Transmission electron microscopy revealed that their substructure exhibited a typical double-layered membrane.Nanoparticle tracking analysis indicated that the size was(164.3±14.0)nm.The result of WB demonstrated that the microparticles expressed specific markers,including TSG101,CD9 and CD41.③ MKMPs were absorbed after being co-cultured with HUVECs for 3 h and enhanced the ability of HUVECs to form tubes and migrate.Notably,the treatment of 5 mg·L-1 MKMPs was more effective than 5 mg·L-1 PMPs treatment.The results of ELISA showed that the content of VEGF from MKMPs was higher than from PMPs,which may be the key factor in regulating endothelial biological function.CONCLUSION MKs derived from hESCs can generate functional microparticles which can promote angiogenesis.

Objective:To develop and evaluate a medical visual question answering (VQA) model for neonatal lung ultrasound (LUS) images to enhance intelligent auxiliary diagnosis of neonatal pulmonary diseases.Methods:Using data from neonates admitted to Beijing Obstetrics and Gynecology Hospital, Capital Medical University (January 2023 to December 2024), an image-question-answer dataset comprising 251 LUS images was constructed [43 pneumonia (17.1%), 42 neonatal respiratory distress syndrome (16.7%), 83 transient tachypnea (33.1%), and 83 normal (33.1%) images] with a four-tier medical question-answer framework. Building upon the Qwen2.5-VL-7B base model and integrating LoRA fine-tuning with chain-of-thought prompting, we developed the NLUS-VQA model to enhance visual-language semantic alignment and enable stepwise clinical reasoning, achieving efficient small-sample adaptation. Model performance was comprehensively assessed through natural language generation metrics (BLEU-4, ROUGE-1/2/L), qualitative evaluation of characteristic recognition, and clinical consistency analysis.Results:(1) Quantitative evaluation demonstrated that NLUS-VQA achieved scores of 22.38 (BLEU-4), 48.26 (ROUGE-1), 22.40 (ROUGE-2), and 37.20 (ROUGE-L), representing significant improvements over baseline models. (2) Qualitatively, the model exhibited strong performance in identifying lung consolidation, coalescent B-lines, and snowflake signs, with its chain-of-thought strategy enhancing clinical interpretability and answer accuracy. (3) Clinically, NLUS-VQA achieved a Cohen's Kappa coefficient of 0.78 and diagnostic accuracy of 80.8% (21/26), indicating substantial agreement with clinical experts.Conclusion:The NLUS-VQA model demonstrates robust interpretability in recognizing key sonographic patterns (e.g. lung consolidation, confluent B-lines, and snowflake signs), providing a scalable framework for small-sample medical image analysis, though diagnostic performance on complex conditions remains limited by dataset scale and minority class representation.

Objective To analyze influencing factors of paclitaxel coated ballon(PCB)on long-term restenosis in patients with severe subpatellar artery lesions in type 2 diabetes mellitus(T2DM),and to construct a prediction model.Methods A total of 268 T2DM patients with severe infra-popliteal artery disease and received PCB treatment were selected.Patients were followed up for 1 year after treatment.Patients with target vessel restenosis were included in the observation group,and the other patients were included in the control group.Clinical data of two groups were analyzed.Multivariable Logistic regression analysis was used to analyze influencing factors of long-term restenosis in T2DM patients with severe infra-knee arterial disease,and a nomogram prediction model was constructed.Results A total of 260 patients(97.00%)completed the follow-up,and the incidence of restenosis was 13.85%(36/260).Both univariate and multivariate Logistic regression analysis showed that age,coexisting coronary heart disease,Trans-Atlantic Inter-Society Consensus(TASC)Ⅱ classification,Fontaine staging,glycosylated hemoglobin(HbA1c)and low density lipoprotein cholesterol(LDL-C)were independent influencing factors for the occurrence of long-term restenosis in T2DM patients with severe infra-popliteal artery disease(P<0.05).The risk factor with the highest score in the constructed nomogram prediction model was HbA1c,followed by age,LDL-C,TASCⅡ classification,Fontaine stage and coronary heart disease.According to the column chart,the total score was 210 points,and the probability of long-term restenosis was 90%.The discrimination of the nomogram model was 0.866,with a Brier score of 0.081 and a calibration slope of 0.733.When the risk threshold was 0.15 to 1.0,the net benefit rate of long-term restenosis in T2DM patients with severe infra-popliteal artery disease was greater than that of individual evaluation.The smaller the risk threshold,the greater the net benefit rate.The benefit was the best when the threshold reached 0.23.Conclusion The influencing factors for long-term restenosis in T2DM patients with severe subknee artery disease treated by PCB include age,combined coronary heart disease,TASCⅡ grade,Fontaine stage,HbA1c and LDL-C.The prediction model based on the above influencing factors has important value in predicting long-term restenosis in patients.

Aim To investigate the inhibition role and mechanism of adipose derived mesenchymal stem cell(ADMSC)exosomes(Exo)on adverse ventricular remodeling after myocardial infarction(MI).Methods The chan-ges of autophagy and inflammasomes phenotype of cardiac fibroblasts after H2O2 treatment were observed.MI rats were in-jected with an equal volume of normal saline,adipose derived mesenchymal stem cell exosomes(MSC-Exo)or fibroblast exosomes(MEF-Exo)via a tail vein.The expression of autophagy related 16 like protein 1(ATG16L1),autophagy re-lated protein 7(ATG7)and NOD-like receptor protein 3(NLRP3),inflammatory response,the degree of myocardial fi-brosis,and the cardiac function were observed in different groups.Results After treatment with H2O2 on cardiac fi-broblasts,the expressions of ATG16L1 and ATG7 were significantly decreased(P＜0.001),NLRP3 was significantly in-creased(P＜0.001),and the levels of inflammatory cytokines interleukin-1β(IL-1β)and IL-18 were significantly elevated(P＜0.001).After MI rats were intervened with MSC-Exo,the expressions of autophagy related proteins ATG16L1 and ATG7 were significantly up-regulated(P＜0.001),NLRP3 was significantly down-regulated(P＜0.001),serum IL-1β and IL-18 levels were significantly decreased(P＜0.001),fibrosis-related proteins collagen Ⅰ and Ⅲ were significantly reduced(P＜0.001),myocardial fibrosis was significantly relieved(P＜0.001),and cardiac function was sig-nificantly improved(P＜0.001).Conclusion Adipose derived MSC-Exo play a role in inhibiting adverse ventricular remodeling after MI by regulating the balance of autophagy and NLRP3 inflammasomes.

This study aims to investigate the synergistic antimicrobial effect of demethylzeylasteral meropenem against NDM-1-positive Escherichia coli.Firstly,the nitrocefin hydrolysis assay and molecular docking assay were used to determine the inhibitory effect and mechanism of demethyl-zeylasteral on the hydrolytic activity of the NDM-1 protein.Secondly,the synergistic bacteriostatic effect of demethylzeylasteral with the carbapenem antibiotic meropenem was confirmed by the checkerboard minimum inhibitory concentration assay,growth curve assay,and inhibition zone as-say.The synergistic bactericidal effect of the combination was further determined by the time-kill-ing assay,live/dead bacterial staining assay,and membrane integrity assay.Finally,the G.mellonel-la infection model demonstrated the protective effect of demethylzeylasteral and meropenem.The results indicated that demethylzeylasteral significantly inhibited the hydrolytic activity of NDM-1(IC50=0.32 mg/L)and competitively affected the binding of NDM-1 to meropenem.In vitro tests showed that demethylzeylasteral had good a synergistic antibacterial effect with meropenem and could exert a synergistic bactericidal effect by enhancing the membrane damage of bacteria caused by meropenem.In vivo assays demonstrated that demethylzeylasteral increased the protective effect of meropenem from 67％to 80％.The results obtained in this study provide a therapeutic strategy and antibacterial synergist for treating NDM-1-positive bacterial infections.

Objective:To analyze the medium- and long-term outcomes of V-Y quadricepsplasty in primary total knee arthroplasty (TKA) to expose an ankylosed or stiff knee joint.Methods:From May 2010 to February 2019, a total of 12 patients with TKA revealed by V-Y quadricepsplasty in West China Hospital of Sichuan University due to knee ankylosis or stiffness were retrospectively analyzed, including 7 males and 5 females, aged (53.9±14.9) years (range, 24 to 72 years), 6 patients on the left side and 6 patients on the right side. Preoperative diagnosis: 7 cases of osteoarthritis, 2 cases of rheumatoid arthritis, 1 case of traumatic arthritis, and 2 cases of haemophilic arthritis. Visual analogue scale (VAS), range of motion, quadriceps muscle strength, Knee Society score (KSS) and postoperative complications were recorded before and after operation.Results:All patients successfully completed the operation and were followed up for 102.2±31.1 months (range, 51-141 months). The operation time was 87.0±15.7 min (range, 73 to 123 min), the intraoperative blood loss was 823.6±237.7 ml (range, 555 to 1 471 ml), and the hospital stay was 13.3±6.3 d (range, 6 to 28 d). Postoperative VAS scores were decreased in all patients, and the difference before and after operation was statistically significant ( F=132.000, P<0.001). The VAS scores at 3 months and the last follow-up were 2.2±0.7 points and 1.2±0.4 points, respectively, lower than those before operation (5.2±0.7 points), and the difference was statistically significant ( P<0.05). KSS knee scores were higher in all patients after operation, and the difference was statistically significant before and after operation ( F=40.960, P<0.001). KSS knee scores at 3 months and the last follow-up were 56.0±14.1 points and 74.3±16.1 points, respectively, higher than those before operation (26.1±7.8 points), and the difference was statistically significant ( P<0.05). Postoperative KSS functional scores were increased in all patients, and the difference before and after operation was statistically significant ( F=24.332, P<0.001). The KSS functional scores at 3 months and the last follow-up were 52.9±19.4 points and 79.2±19.6 points, respectively, higher than those before operation (27.1±15.6 points), and the difference was statistically significant ( P<0.05). Postoperative knee joint motion was increased in all patients, and the difference was statistically significant before and after operation ( F=24.145, P<0.001). The range of motion of the knee joint at 3 months and the last follow-up was 57.5°±22.2° and 70.0°±25.9°, respectively, which was higher than the preoperative 12.5°±14.1°, and the difference was statistically significant ( P<0.05). Preoperative quadriceps muscle strength was grade 3 in 2 cases and grade 4 in 10 cases; at the last follow-up, grade 4 in 1 case and grade 5 in 11 cases, and the muscle strength was improved compared with that before operation, the difference was statistically significant ( Z=11.000, P<0.001). At the last follow-up, there were no complications such as wound seepage, delayed healing, superficial or deep soft tissue infection, periprosthesis infection and loosening, deep vein thrombosis and pulmonary embolism. Conclusion:In patients with ankylosed or stiff knee receiving TKA, the use of V-Y quadricepsplasty can increase the exposure, thereby improving the range of knee motion and quadriceps muscle strength.