Background: Murraya, a genus of shrubs and trees in the Rutaceae family, consists of approximately 9 species in China with significant medicinal and horticultural value. However, the phylogeny and taxonomy of Murraya species remain controversial, particularly with respect to Murraya exotica and M. paniculata. Objective: This study aimed to provide insights into the taxonomy, phylogeny, and identification of Murraya. Methods: In this study, the chloroplast (CP) genomes of 7 Murraya species were sequenced, assembled, and subjected to comparative and phylogenetic analyses. Results: The CP genomes of Murraya ranged from 158,573 to 160,817 bp in length and encoded 112 unique genes, including 78 protein-coding genes, 30 tRNA genes, and 4 rRNA genes. Similar to other angiosperms, the inverted repeat regions of the CP genomes exhibited lower sequence divergence than the single-copy regions, and coding regions were more conserved than noncoding regions. Comparative analysis identified several highly variable regions (eg, matK, ycf1, ndhI-ndhA, trnH-GUG-psbA, rpl32-trnL) that could serve as molecular markers for species identification in Murraya. Among these, the ycf1 gene was validated as a useful marker for distinguishing M. exotica from M. paniculata. Positive selection was detected in 10 genes, including rbcL, psaJ, ndhD, ndhF, rpl2, rpl20, ycf1, accD, ccsA, and rpl32. Phylogenetic analysis based on CP genomes supported the recognition of M. exotica and M. paniculata as independent species. Moreover, the phylogenetic trees indicated that Murraya is not monophyletic, with sect. Bergera showing a closer relationship to Clausena. Molecular dating results suggested that the diversification of M. paniculata, M. alata, and M. exotica occurred approximately 9.11 Mya (95% highest posterior density: 4.90-13.87 Mya). Conclusion: These findings provide valuable CP genome data for clarifying the phylogenetic relationships between M. exotica and M. paniculata, and for advancing the study of DNA markers and the evolutionary history of Murraya.

The biliary microbiota is an important component of the biliary system, the structure changes and dysbiosis of the biliary microbiota are closely related to the occurrence and development of biliary diseases. Recent studies have found that the dysbiosis of the biliary microbiota can lead to the occurrence of various biliary diseases such as cholelithiasis, cholangitis, and biliary tract cancer. However, the specific mechanisms of some of these diseases remain unclear. In addition, the alternation of biliary microbiota may also lead to the occurrence of postoperative complications of biliary diseases, such as the risk of infection increased after laparoscopic cholecystectomy (LC) and endoscopic retrograde cholangiopancreatography (ERCP). Although some studies have begun to explore the connection between biliary microbiota and biliary diseases, most research have mainly focused on a single disease type or specific microorganisms. There has been no comprehensive and systematic review of the overall changes in biliary microbiota across different diseases and their association with post biliary surgery infection.Therefore, this article presents a systematic review of current advances linking the alternations of biliary microbiota to the development of biliary diseases and their postoperative infections, as well as the changes in the biliary microbiota before and after various biliary diseases and their surgeries, aiming to provide insightful methods for the early diagnosis, prevention and treatment of biliary diseases and management of postoperative infections.

OBJECTIVE To investigate the effects of Ephedra-Cinnamomum couplet medicinals on respiratory function and airway inflammation in rats with asthma of cold-fluid retention in lung syndrome and its mechanism. METHODS Sixty Wistar rats were randomly divided into blank group， model group， dexamethasone group [1 mg/（kg·d）]， and Ephedra-Cinnamomum low-， medium-， high-dose groups [0.234， 0.936， 1.872 g/（kg·d）]， with 10 rats in each group. The model and treatment groups were sensitized by intraperitoneal injection of antigen solution （ovalbumin 100 mg + aluminum hydroxide 100 mg） and challenged with 1% ovalbumin nebulization， along with exposure to a cold environment and ingestion of cold water， to establish the asthma model with cold-fluid retention in lung syndrome. From day 2， rats received corresponding drugs or normal saline intragastrically， once a day， for 21 consecutive days. The general behavioral changes in each group of rats were observed during the experimental process. The lung function parameters [peak expiratory flow （PEF）， airway resistance （Raw）， functional residual capacity （FRC）， expiratory flow at 50% of forced vital capacity （EF50%）] Δ 基金项目 国家自然科学基金青年科学基金项目（No.82004233）； were measured before modeling and after the last medication as well as serum contents of interleukin-6 （IL-6）， IL-13， tumor necrosis factor- α （TNF- α） and interferon-gamma （IFN- γ） after the last medication were determined； the histopathological morphological changes in the lung tissues of rats were also observed； mRNA expressions of IL-6， IL-3， TNF-α， IFN-γ， thymic stromal lymphopoietin （TSLP）， and Toll-like receptor 4 （TLR4）， as well as protein expressions of TSLP and TLR4 were determined in lung tissue. RESULTS Compared with model group， the lung tissue damage of rats was relieved significantly； Raw， FRC， the contents and mRNA expression levels of IL-6， IL-13 and TNF-α， as well as the mRNA and protein expressions of TSLP and TLR4 were significantly decreased （P＜0.05）， while the contents and mRNA expressions of PEF， EF50 % and IFN-γ were significantly increased in the dexamethasone group and Ephedra-Cinnamomum medium- and high-dose groups （P＜0.05）. Moreover， only a few rats in the two groups exhibited typical symptoms of asthma. CONCLUSIONS Ephedra-Cinnamomum couplet medicinals improve respiratory function and ameliorate airway inflammation in asthma rats with cold-fluid retention in lung syndrome， the mechanism of which may be associated with inhibiting TSLP/TLR4 signaling pathway and modulating Th1/Th2 imbalance.

Objective The complex evolutionary history of the origin of medicinal blue herbs might result in the presence of adulterants, affecting the accuracy and safety of clinical medication. To provide a reference basis for the identification of medicinal blue herbs with leaves as the medicinal part, based on the leaf epidermis microstructure. Methods The species belonging to medicinal blue herbs and their close relatives (10 species from 4 genera) were systematically investigated. The leaf epidermis microstructure of these species was observed and analyzed by optical microscopy and scanning electron microscopy. A species retrieval table was established based on the microstructure characteristics. Results By combining the distribution of stomata, types of subsidiary cells, stomatal index, stomatal density, characteristics of the periclinal walls of epidermal cells, and epidermal appendages, the species Clerodendrum cyrtophyllum, Polygonum tinctorium, Isatis indigotica, I. violascens, I. costata, I. minima, Strobilanthes wallichii, S. dalzielii, S. pentstemonoides, and S. cusia can be accurately distinguished. Conclusion Microscopic characteristics of leaf epidermis can provide reference data for accurately differentiating the botanical origins of medicinal blue herbs.

Objective:To establish a model based on the characteristics of breast cancer ultrasound images through deep learning methods to predict the risk of axillary lymph node metastasis（ALNM）in patients with breast cancer（maximum diameter ≤2.5 cm）before surgery.Methods:A total of 419 patients（3 433 breast tumor ultrasound images）with breast cancer（maximum diameter ≤2.5 cm）who underwent axillary lymph node dissection at Beijing Tiantan Hospital，Capital Medical University from January 2019 to December 2024 were retrospectively included. According to the pathological results of axillary lymph nodes，they were divided into 220 cases in the ALNM occurrence group（positive group）and 199 cases in the non-ALNM occurrence group（negative group）. The breast cancer ultrasound images of the two groups of cases were randomly classified into the training set（2 404 images），the validation set（687 images）and the test set（342 images）according to a ratio of 7∶2∶1. YOLOv8 was used as the basic model of You Only Look Once（YOLO）and optimized. The optimized model was applied to locate and capture the potential ultrasound features of breast cancer cases in the training set. A prediction model was constructed based on the captured ultrasound features. The model was adjusted and optimized through the validation set，and then matched with the case images in the test set. The confusion classification matrix graph and the curve graph for measuring the model performance were used to evaluate the model prediction performance and interpret the model，and the efficacy of this model in identifying breast cancer patients at risk of ALNM was analyzed.Results:There were statistically significant differences between the positive and negative groups in terms of the pathological maximum diameter of breast tumors，pathological T staging，the differentiation degree，the presence of distant metastasis，the maximum diameter measured by ultrasound，the quadrant of breast tumor occurrence，the Breast Imaging - Reporting and Data System（BI-RADS）classification of breast tumors，and the presence of abnormal ultrasound features of lymph node（all P<0.05）. The established deep learning model could automatically perform bounding box localization for the breast cancer of patients.The breast tumors in the positive group had potential ultrasound features that could be captured by the model compared with those in the negative group. The mean average precision（mAP）50 was 0.883，mAP 50-95 was 0.636，PR-AUC was 0.884 5，strict PR-AUC was 0.636 4，the sensitivity was 90.5%，and the specificity was 91.2%，and it had a good predictive efficacy. Conclusions:This prediction model based on the ultrasound characteristics of breast cancer through deep learning can effectively predict breast cancer（maximum diameter ≤ 2.5 cm）with the risk of ALNM，providing an effective basis for the clinical management of axillary lymph nodes in breast cancer patients.

Objective To investigate whether miR-15b-5p can alleviate airway inflammation in asthma by negatively regulating ubiquitin specific peptidase 9X (USP9X) to down-regulate the expression of phosphatidylinositol 4, 5-diphosphate 3-kinase catalytic subunit α/AKT serine/threonine kinase 1 (PIK3CA/AKT1) pathway. Methods USP9X was predicted to be a direct target of miR-15b-5p by using an online database (miRWalk), and the luciferase reporter gene assay was performed to verify it. Co-immunoprecipitation (CO-IP) was used to verify the direct binding between USP9X and PIK3CA and the role of USP9X and its small molecule inhibitor WP1130 in the deubiquitination of PIK3CA. C57 mice were randomly divided into Control group, OVA group, OVA combined with NC group and miR-15b-5p agomir group, with 10 mice in each group. BEAS-2B cells were induced with interleukin 13 (IL-13) and treated with miR-15b-5p mimic. HE, Masson, PAS, immunohistochemistry, immunofluorescence staining, flow cytometry, Western blot and quantitative real-time PCR(qRT-PCR) were performed. Results It was found that the administration of miR-15b-5p agomir and mimic could reduce peribronchial inflammatory cells and improve airway inflammation, and miR-15b-5p could target negative regulation of USP9X. USP9X could directly bind to PIK3CA and regulate PIK3CA level in a proteasome-dependent manner, and USP9X could deubiquitinate K29-linked PIK3CA protein. Down-regulation of USP9X could increase PIK3CA ubiquitination level. WP1130, a small molecule inhibitor of USP9X, has the same effect as knockdown of USP9X, both of which could increase the ubiquitination level of PIK3CA and reduce the protein level of PIK3CA. Conclusion The miR-15b-5p/USP9X/PIK3CA/AKT1 signaling pathway may provide potential therapeutic targets for asthma.
Animals ; MicroRNAs/metabolism* ; Asthma/pathology* ; Class I Phosphatidylinositol 3-Kinases/genetics* ; Ubiquitin Thiolesterase/metabolism* ; Proto-Oncogene Proteins c-akt/genetics* ; Mice ; Signal Transduction ; Mice, Inbred C57BL ; Humans ; Inflammation/genetics* ; Cell Line ; Female ; Male

Objective To investigate the effects of Yishen Zhuyu Tongluo Decoction on chondrocyte pyroptosis and ferroptosis in rats with knee osteoarthritis(KOA).Methods Forty SD were divided randomly into a sham operation group(n=10)and a modeling group(n=30).A KOA model was established in the modeling group by anterior cruciate ligament transection of the right hind limb knee joint.The drawer test was used to confirm the successful establishment of the model.Post-surgery,the rats were subjected to 30 min of forced activity daily to induce KOA.Lameness and hopping movements were observed after 4 weeks,and hematoxylin and eosin staining confirmed cartilage surface damage,deformation,and inflammatory cell infiltration,indicating successful modeling.The model rats were then assigned randomly to a model group(n=8),celecoxib group(n=8),and Yishen Zhuyu Tongluo Decoction group(n=8).The sham operation and model groups received 10 mL/kg of saline by gavage,the celecoxib group received 12 mg/kg of celecoxib solution,and the Yishen Zhuyu Tongluo Decoction group received 5.4 g/kg of the herbal decoction,once daily for 8 weeks.After the interventions,the rats were anesthetized,blood was collected from the abdominal aorta and serum was separated,and the knee joints were isolated.Three samples were fixed in paraformaldehyde,while the remaining cartilage tissue was reserved.Pathological changes in joint cartilage were observed by hematoxylin and eosin and Safranin O-Fast Green staining.Bone microstructure was analyzed using micro-computed tomography.Serum interleukin(IL)-1β and IL-18 protein levels were detected by enzyme-linked immunosorbent assay.Relative mRNA expression levels of NLRP3,ASC,Caspase-1,GSDMD,IL-1β,IL-18,ACSL-4,FTH-1,GPX-4,and cyclooxygenase(COX)-2 were detected by reverse transcription quantitative real-time PCR,and protein expression of NLRP3,Caspase-1,and COX-2 were detected by immunohistochemistry.Results Compared with the sham operation group,model rats showed surface damage and deformation of cartilage tissue,disordered cell arrangement in all layers,significant loss of Safranin O-Fast Green staining,and sparse and irregular trabecular bone distribution.Serum levels of the inflammatory factors IL-1β and IL-18 were elevated(P＜0.01).mRNA expression levels of NLRP3,ASC,Caspase-1,GSDMD,IL-1β,IL-18,ACSL-4,and COX-2 were also significantly increased(all P＜0.01),while FTH-1 and GPX-4 mRNA expression levels were decreased(P＜0.01).Compared with the model group,rats in the celecoxib and Yishen Zhuyu Tongluo Decoction groups showed smoother and more intact cartilage surfaces,significantly increased cell counts,less loss of Safranin O-Fast Green staining,denser trabecular bone,and thicker cortical bone,with improved bone microstructure.Serum IL-1β and IL-18 levels were reduced(P＜0.01),NLRP3(P＜0.01,P＜0.05),ASC,Capase-1,GSDMD,IL-1β,IL-18,ACSL-4,and COX-2 mRNA levels were decreased(all P＜0.01),and relative expression levels of FTH-1 and GPX-4 mRNA were significantly increased(P＜0.01).Conclusions Yishen Zhuyu Tongluo Decoction can treat KOA by inhibiting chondroptosis and ferroptosis in chondrocytes,reducing serum IL-1β and IL-18 levels,and improving bone microstructure.

In recent years,with the continuous advancement of deep space exploration missions,astronauts have been increasingly exposed to complex and extreme environmental factors in space,such as microgravity,space radiation,circadian rhythm disruption,and confined isolation.These conditions can easily induce brain dysfunction in astronauts,manifesting as cognitive decline,emotional disturbances,sleep disorders,and neuroinflammatory responses.Traditional Chinese Medicine(TCM),characterized by its holistic regulation and syndrome differentiation-based treatment principles,exhibits multi-target regulation and systemic coordination,and has attracted growing attention in the field of brain function protection.This study systematically reviews the protective applications and research progress of TCM and related techniques in both actual spaceflight missions and simulated space environment models against brain functional disorders.The underlying mechanisms and future prospects are discussed,with the aim of providing insights for safeguarding astronauts'brain health and laying a theoretical foundation for the precise intervention and systematic protection strategies of TCM in future deep space missions.

Objective To evaluate the preliminary effects of a Traditronal Chinese Medicine(TCM)incentive policy aimed at the number of discharge patients on the synergistic improvement of performance assessment indicators in the Traditional Chinese Hospital.Methods It selected detailed data from the first page of medical records at the grade A teritary hospital from October,2018 to October,2021.Using the Interrupted Time Series analysis method,two breakpoints were taken as the start time of COVID-19(December 2019)and the official implementation time of dis-charge performance program(September 2020).The utilization rate of TCM decoction pieces among discharged pa-tients,the utilization rate of TCM non-drug therapy among discharged patients,the proportion of discharged pa-tients mainly treated with TCM,and the proportion of inpatient medical service income before and after the break-point were analyzed.Results Despite the impact of COVID-19,after the policy was implemented,four key indicators showed a positive growth trend:the proportion of discharged patients treated primarily with TCM,the rate of TCM decoction use among discharged patients,the rate of TCM non-pharmaceutical therapy use among discharged pa-tients,and the proportion of medical service income,respectively.Conclusion The introduction of a TCM reward module in the discharge performance bonus plan of the sample hospital effectively stimulated the enthusiasm of medical staff in applying TCM.The utilization rate of TCM decoction slices,the utilization rate of TCM non-drug therapy,the proportion of discharged patients treated mainly by TCM and the proportion of medical service income all showed an upward trend,indicating that the incentive policy of TCM has a positive impact on the national examination index of TCM hospitals.

Objective To investigate the association between plasma homocysteine(Hcy)level and coronary artery calcification(CAC)and its clinical predictive efficacy.Methods A total of 172 patients who underwent coronary CT angiography North China University of Science and Technology Affiliated Hospital from April 2019 to May 2021 and CAC score(CACS)＞0 were enrolled.According to the CACS value,the subjects were divided into mild calcification group(n=136)and severe calcification group(n=36),and the clinical characteristics of two groups were compared and analyzed.Multivariate Logistic regression model was used to screen the independent influencing factors of CAC severity,and a prediction model was constructed based on the Hcy detection value.The clinical diagnostic value was evaluated by the receiver operating characteristic(ROC)curve.Results There were significant differences in Hcy,white blood cell count,triglyceride and magnesium ion levels between two groups(P＜0.05).Multivariate Logistic regression analysis showed that Hcy,white blood cell count and magnesium ion level were independent risk factors for the progression of CAC.Furthermore,a regression model based on Hcy was constructed and ROC curve was fitted to evaluate its predictive efficacy.The results suggested that the predictive model had the best performance when the critical value of Hcy was set at 27.4μmol/L:the sensitivity was 55.6％,the specificity was 97.1％,and the area under the curve was 0.765.Conclusion Hcy serves as an independent risk factor for the severity of CAC and can effectively predict the progression of CAC with high accuracy.