ObjectiveTo investigate the mechanism by which Shengjiang Pingchuan Zhike pills （SPZP） alleviate airway injury in the rat model of cough variant asthma （CVA） via regulating autophagy through activating the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/Unc-51-like autophagy activating kinase 1 （ULK1） signaling pathway based on transcriptomics， thus providing experimental evidence for the clinical application of SPZP. MethodsForty SPF-grade SD rats were enrolled in the present study and randomly via the random number table method assigned into four groups： a blank group， a model group， a SPZP （2 160 mg·kg^-1·d^-1） group， and a positive drug （montelukast sodium， MTL， 0.9 mg·kg^-1·d^-1）group， with 10 rats in each group. After one week of adaptive feeding， the rat model of cough variant asthma （CVA） was established by two-week sensitization with ovalbumin （OVA） and aluminium hydroxide， followed by capsaicin atomization challenge. Rats in the blank group received an equal volume of normal saline during modeling. Upon successful modeling， rats in the SPZP group and positive drug group were administrated with SPZP suspension and MTL solution， respectively， while those in the blank and model groups received the same dose of normal saline by gavage. The whole intervention lasted for 21 consecutive days. After the intervention， general behaviors of rats in each group were observed. Pulmonary function indicators were subsequently detected. Hematoxylin-eosin （HE） staining was performed to visualize the pathological morphology of the lung tissue， and periodic acid-Schiff （PAS） staining was adopted to evaluate airway mucus secretion. Transcriptomic sequencing was conducted to screen differentially expressed genes in the lung tissue across groups. The levels of inflammatory cytokines including interleukin （IL）-4， IL-5， immunoglobulin E （IgE）， and interferon-γ （IFN-γ） in the bronchoalveolar lavage fluid （BALF） were quantified by enzyme-linked immunosorbent assay （ELISA）. Immunofluorescence staining was used for detecting the co-localization of autophagy-related protein Beclin-1 and phosphorylated mammalian target of rapamycin （p-mTOR）. Transmission electron microscopy was employed to count autophagosomes and observe mitochondrial morphological changes in the lung tissue. Western blot assay was utilized to measure the expression levels of AMPK/mTOR/ULK1 axis-related proteins， including AMPK， phosphorylated adenosine monophosphate-activated protein kinase （p-AMPK）， mTOR， phosphorylated mammalian target of rapamycin （p-mTOR）， ULK1， and phosphorylated Unc-51-like autophagy activating kinase 1 （p-ULK1） as well as the autophagy biomarkers Beclin-1 and selective autophagy adaptor protein p62 in lung specimens. ResultsCompared with the blank group， the model group exhibited markedly elevated cough frequency accompanied by wheezing and stridor， dull fur， poor mental activity， diminished motor activity， accelerated respiratory rate， and weakened sensory response and mobility to external stimuli， accompanied by a declining tendency of body mass. Meanwhile， the model group exhibited decreased dynamic lung compliance， increased airway resistance （P<0.01）， severe pathological damage and excessive mucus secretion in the lung tissue （P<0.01）， and elevated levels of IL-4， IL-5， and IgE and declined level of IFN-γ in the BALF （P<0.01）. Ultrastructural observation revealed damaged mitochondrial morphology and a sharp reduction in the number of autophagosomes in the lung tissue. In addition， the expression of p-AMPK， p-ULK1， and Beclin-1 were downregulated， whereas p-mTOR and p62 were highly expressed （P<0.05， P<0.01） in the model group. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses demonstrated significant enrichment of AMPK， mTOR， and autophagy-related pathways. Compared with the model group， both the positive drug group and SPZP group obtained visible improvements in general behavioral manifestations and obvious recovery of pulmonary function indicators （P<0.05）， alleviated pulmonary pathological lesions and suppressed excessive airway mucus secretion （P<0.05）， recovered levels of inflammatory cytokines in the BALF （P<0.05）， restored mitochondrial morphology， increased autophagosomes， upregulated expression levels of p-AMPK， p-ULK1， and Beclin-1， and downregulated expression levels of p-mTOR and p62 （P<0.05）. ConclusionSPZP can effectively ameliorate airway inflammation and improve the pulmonary function in the rat model of CVA by activating the AMPK/mTOR/ULK1 signaling pathway， enhancing autophagy in the lung tissue， and inhibiting excessive airway mucus secretion and immune inflammatory imbalance.

Objective: To explore new therapeutic strategies for periodontitis, this study prepared extracellular vesicles (EVs) derived from human umbilical cord mesenchymal stem cells (hUC-MSCs) loaded with saikosaponin D (SSD) and evaluated their efficacy in a mouse model of periodontitis. Methods: This study was approved by the Medical Ethics Committee and the Laboratory Animal Ethics Committee of the unit. hUC-MSCs at logarithmic growth phase were seeded into 96 well plates and then treated with various concentrations of SSD for 24 hours. The CCK-8 assay was used to determine the SSD concentration that preserves hUC-MSC viability. Flow cytometry (FCM) assessed SSD-induced apoptosis in hUC-MSCs and determined drug loading and loading efficiency. Nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM) characterized the morphology and size of hUC-MSCs-EVs and hUC-MSCs-SSD-EVs. C57BL/6J mice were divided into five groups with six mice per group: control group, periodontitis model group (PD), PD+SSD group (2 mg/kg), PD+hUC-MSCs-EVs group (10 μg/μL) and PD+hUC-MSCs-SSD-EVs group (10 μg/μL). Administration was performed 3 times a week for 4 consecutive weeks. Micro-CT and hematoxylin-eosin (HE) staining evaluated alveolar bone resorption in periodontal tissues of a mouse model of periodontitis. RT-qPCR measured inflammatory cytokine expression in periodontal tissues of a mouse model of periodontitis. Immunofluorescence analyzed macrophage M1/M2 polarization and runt-related transcription factor 2 (RUNX2) protein expression in periodontal tissues of a mouse model of periodontitis. Results: The CCK-8 assay showed that 10 μmol/L SSD had little effect on hUC-MSCs viability (P>0.05). FCM confirmed the successful loading of SSD into EVs with a loading efficiency of 11.40%. TEM and NTA revealed that hUC-MSCs-SSD-EVs presented round or oval membranous structures with diameters ranging from 200 to 300 nm. Micro CT and HE staining showed that, compared with the PD group, the distance from cementoenamel junction to alveolar bone crest (CEJ-ABC) was markedly shortened (P<0.01）.The alveolar bone of periodontitis‑model bone volume fraction (BV/TV) and trabecular number (Tb.N) were significantly elevated in the PD+hUC-MSCs-SSD-EVs group (P<0.01). RT-qPCR and IF results showed that hUC-MSCs-SSD-EVs in periodontal tissues markedly downregulated the expression of pro-inflammatory cytokines including interleukin-1β, interleukin-6, tumor necrosis factor-α, interleukin-12 and interleukin-22 (P<0.01), upregulated the expression of the anti-inflammatory factor interleukin-10 (P<0.01), promoted the polarization of macrophages from the M1 phenotype to the M2 phenotype, and elevated the protein expression level of RUNX2 (P<0.01). Conclusion hUC-MSC-SSD-EVs effectively alleviated inflammatory responses in a mouse model of maxillary periodontitis and reduced alveolar bone loss, providing a new idea for the treatment of periodontitis.

Guidelines for Digital Ancient Books of TCM Indexing(T/CIATCM 119-2024)is based on the theoretical knowledge,disciplinary methods,and practical applications of TCM classical cataloging.Taking digital ancient books of TCM as the object,it systematically reveals the content of TCM knowledge,which is an essential indexing processing standard for building an intelligent retrieval system for TCM ancient books,and can provide support for the deep development and innovative utilization of TCM knowledge.It can not only promote the co-construction and sharing of ancient book resources in the TCM industry,but also promote the standardization construction and application of TCM information.This standard specifies the principles,methods,and examples of free indexing of digital ancient books of TCM based on their original content.It is applicable to the indexing and processing of digital ancient books of TCM for TCM professional libraries and related institutions,and to the data processing and construction of various types of TCM ancient book databases.

Objective To observe the efficacy of a nomogram based on conventional ultrasonic manifestations for predicting malignant risk of breast non-mass lesions(NML).Methods A total of 120 NML diagnosed with ultrasound in 113 female patients were retrospectively included and divided into malignant group(n=44)and benign group(n=76)based on pathological results.Patients' age,ultrasonic manifestations of NML and ipsilateral axillary lymph node were compared between groups.Then a nomogram was constructed using a multivariable logistic regression model,and its diagnostic performance and accuracy were validated.Results The ratios of patients' aged≥45 years,with abnormal ipsilateral axillary lymph nodes,NML with maximum diameter≥2 cm,posterior echo attenuation,rich blood flow signals,architectural distortion and microcalcification were significantly different between groups(all P＜0.05).Patient's age≥45 years,with abnormal ipsilateral axillary lymph nodes,and NML with rich blood flow signals,architectural distortion and microcalcification were all independent predictive factors for high malignant risk of breast NML(all P＜0.05).The constructed nomogram achieved an area under the receiver operating characteristic curve of 0.901 for predicting malignant risk of breast NML,and its predicting probabilities demonstrated good agreement with the actual probabilities(Hosmer-Lemeshow test P＞0.05).Conclusion The nomogram incorporating patient's age,ultrasonic manifestations of NML and ipsilateral axillary lymph node could effectively quantify malignancy risk of breast NML.

Objective:To explore the value of nomogram model based on multimodal ultrasound features for predicting the malignant risk of micro lesions in breast areola region.Methods:The case data of Beijing Tiantan Hospital affiliated to Capital Medical University from May 2020 to July 2024 were retrospectively analyzed. A total of 50 patients with benign intraductal papilloma（bIDP group）and 54 patients with malignant risk breast tumor（mrBT group）were found to have micro lesions in breast areola region and confirmed by puncture or surgical pathology. Clinical data，conventional ultrasound and contrast-enhanced ultrasound features were compared between the two groups. Multivariate Logistic regression analysis and Lasso regression analysis were performed on statistically significant factors to screen out influencing factors. ROC curves were plotted to evaluate diagnostic efficacy，nomogram model and clinical decision curves were constructed to evaluate clinical benefits.Results:The differences of age，nipple discharge presentation，conventional ultrasound features（including boundary，morphology，aspect ratio，internal echo，internal microcalcification，far-field echo，peripheral irregular hyperechoic ring，dilate of peripheral ducts），and contrast-enhanced ultrasound features（including wash-in time，enhancement intensity，enhancement mode，enhancement scope，blood perfusion defect，crab foot sign，penetrating vessels）were statistically significant between the bIDP group and mrBTgroup（all P<0.05）. Regression analysis showed that age，uniformity of internal echo within the lesion，dilation of surrounding ducts，and enhanced crab foot sign were the affect factors for the diagnosis of mrBT（all P<0.05）. Based on these factors，a nomogram model was constructed with an area under ROC curve（AUC）of 0.907（95% CI=0.851-0.963），a sensitivity of 0.907，and a specificity of 0.780. The decision curve analysis showed that the collective model had good predictive performance. Conclusions:The nomogram model based on multimodal ultrasound features has good value in predicting malignant risk micro breast tumor of areola region.

Guidelines for Digital Ancient Books of TCM Indexing(T/CIATCM 119-2024)is based on the theoretical knowledge,disciplinary methods,and practical applications of TCM classical cataloging.Taking digital ancient books of TCM as the object,it systematically reveals the content of TCM knowledge,which is an essential indexing processing standard for building an intelligent retrieval system for TCM ancient books,and can provide support for the deep development and innovative utilization of TCM knowledge.It can not only promote the co-construction and sharing of ancient book resources in the TCM industry,but also promote the standardization construction and application of TCM information.This standard specifies the principles,methods,and examples of free indexing of digital ancient books of TCM based on their original content.It is applicable to the indexing and processing of digital ancient books of TCM for TCM professional libraries and related institutions,and to the data processing and construction of various types of TCM ancient book databases.

Objective:To explore the value of nomogram model based on multimodal ultrasound features for predicting the malignant risk of micro lesions in breast areola region.Methods:The case data of Beijing Tiantan Hospital affiliated to Capital Medical University from May 2020 to July 2024 were retrospectively analyzed. A total of 50 patients with benign intraductal papilloma（bIDP group）and 54 patients with malignant risk breast tumor（mrBT group）were found to have micro lesions in breast areola region and confirmed by puncture or surgical pathology. Clinical data，conventional ultrasound and contrast-enhanced ultrasound features were compared between the two groups. Multivariate Logistic regression analysis and Lasso regression analysis were performed on statistically significant factors to screen out influencing factors. ROC curves were plotted to evaluate diagnostic efficacy，nomogram model and clinical decision curves were constructed to evaluate clinical benefits.Results:The differences of age，nipple discharge presentation，conventional ultrasound features（including boundary，morphology，aspect ratio，internal echo，internal microcalcification，far-field echo，peripheral irregular hyperechoic ring，dilate of peripheral ducts），and contrast-enhanced ultrasound features（including wash-in time，enhancement intensity，enhancement mode，enhancement scope，blood perfusion defect，crab foot sign，penetrating vessels）were statistically significant between the bIDP group and mrBTgroup（all P<0.05）. Regression analysis showed that age，uniformity of internal echo within the lesion，dilation of surrounding ducts，and enhanced crab foot sign were the affect factors for the diagnosis of mrBT（all P<0.05）. Based on these factors，a nomogram model was constructed with an area under ROC curve（AUC）of 0.907（95% CI=0.851-0.963），a sensitivity of 0.907，and a specificity of 0.780. The decision curve analysis showed that the collective model had good predictive performance. Conclusions:The nomogram model based on multimodal ultrasound features has good value in predicting malignant risk micro breast tumor of areola region.

Objective To observe the efficacy of a nomogram based on conventional ultrasonic manifestations for predicting malignant risk of breast non-mass lesions(NML).Methods A total of 120 NML diagnosed with ultrasound in 113 female patients were retrospectively included and divided into malignant group(n=44)and benign group(n=76)based on pathological results.Patients' age,ultrasonic manifestations of NML and ipsilateral axillary lymph node were compared between groups.Then a nomogram was constructed using a multivariable logistic regression model,and its diagnostic performance and accuracy were validated.Results The ratios of patients' aged≥45 years,with abnormal ipsilateral axillary lymph nodes,NML with maximum diameter≥2 cm,posterior echo attenuation,rich blood flow signals,architectural distortion and microcalcification were significantly different between groups(all P＜0.05).Patient's age≥45 years,with abnormal ipsilateral axillary lymph nodes,and NML with rich blood flow signals,architectural distortion and microcalcification were all independent predictive factors for high malignant risk of breast NML(all P＜0.05).The constructed nomogram achieved an area under the receiver operating characteristic curve of 0.901 for predicting malignant risk of breast NML,and its predicting probabilities demonstrated good agreement with the actual probabilities(Hosmer-Lemeshow test P＞0.05).Conclusion The nomogram incorporating patient's age,ultrasonic manifestations of NML and ipsilateral axillary lymph node could effectively quantify malignancy risk of breast NML.

BACKGROUND:Piezo1,a mechanosensitive protein,is tightly connected to osteogenic differentiation,and it has been demonstrated that TAZ has a role in regulating osteogenic differentiation.It is unclear whether TAZ participates in the regulation of osteogenic differentiation of human bone marrow mesenchymal stem cells by Piezo1,so it is crucial to investigate its unique mechanism to prevent osteonecrosis of the femoral head. OBJECTIVE:To elucidate what function Piezo1 plays in osteogenic differentiation and TAZ expression in human bone marrow mesenchymal stem cells. METHODS:The siRNA targeting Piezo1 was constructed and transfected into 293T cells.The silencing efficiency was detected by RT-qPCR.The selected Piezo1-Home-2337 was packaged according to the silencing efficiency,and its optimal multiplicity of infection value was assayed by immunofluorescence staining.The packaged Piezo1 silencing recombinant lentivirus was transfected into human bone marrow mesenchymal stem cells,and its silencing effect was detected by RT-qPCR and western blot assay.Alizarin red staining,alkaline phosphatase activity analysis,immunofluorescence staining,RT-qPCR and western blot assay were utilized to analyze the effect of silencing Piezo1 on the osteogenic differentiation of human bone marrow mesenchymal stem cells. RESULTS AND CONCLUSION:(1)The mRNA and protein levels of Piezo1 in human bone marrow mesenchymal stem cells transfected by si-Piezo1 were decreased significantly,with a statistically significant difference compared with normal and negative control groups.(2)The alkaline phosphatase activity in the si-Piezo1 group was much lower and the calcium deposition in the si-Piezo1 group was significantly reduced compared with the negative control group.(3)The mRNA levels of osteogenesis-related genes including Runt-related transcription factor 2(Runx2),osteopontin(OPN),distal-less homeobox 5(DLX5),osteocalcin,β-catenin and Tafazzin(TAZ)in the si-Piezo1 group were significantly decreased compared with the negative control group.Afterward,the expression levels of TAZ and β-catenin protein in the si-Piezo1 group were down-regulated significantly compared with the negative control group,whereas the expression levels of p-TAZ and p-β-catenin protein in the si-Piezo1 group had the opposite condition.(4)The results of immunofluorescence staining showed that the expression of TAZ and β-catenin in human bone marrow mesenchymal stem cells in the si-Piezo1 group was less compared with the negative control group.(5)These findings indicate that Piezo1 can promote the osteogenic differentiation of human bone marrow mesenchymal stem cells.The osteogenic ability of human bone marrow mesenchymal stem cells is significantly reduced after silencing Piezo1,and the expression of TAZ is also reduced.