ObjectiveThis paper aims to find the identified and validated clinical biomarker data building upon a clinical study of early-phase phase Ⅱ and investigate the correlation analysis of Huanglian Jiedu Wan on syndrome improvement and clinical biomarkers in the treatment of "excess heat-toxicity" based on a machine learning model. Additionally， the effective prediction of clinical biomarker values for the main symptoms of the "excess heat-toxicity" syndrome was assessed. MethodsA total of 229 patients meeting the inclusion criteria for "excess heat-toxicity" syndrome were randomly divided into the Huanglian Jiedu Wan group and the placebo group. Syndrome score transition matrices were constructed for the Huanglian Jiedu Wan group and the placebo group based on three main symptoms of "excess heat-toxicity" syndrome， such as oral ulcers， sore throat， and gum swelling and pain. Data from the patients with these three syndromes were also integrated for an overall analysis. The corresponding syndrome score transition matrices were further constructed to visualize symptom change trends of the patients in the two groups via heatmaps. Based on the identified and validated clinical biomarkers related to inflammation， oxidative stress， and energy metabolism in the early phase， Spearman correlation analysis was employed to analyze and evaluate the associations between clinical biomarkers and syndrome improvement. Key clinical biomarkers reflecting the effect of Huanglian Jiedu Wan were screened through the comparison of differences between groups. An extreme gradient boosting （XGBoost） algorithm was used to develop a prediction model for main symptom classification， with classification performance evaluated through 10-fold cross-validation. Feature importance analysis was applied to identify variables with the greatest contribution to the prediction result. ResultsThe syndrome transition matrix results indicated that the Huanglian Jiedu Wan group showed a superior effect to the placebo group in improving oral ulcers， sore throat， and overall symptoms， with significant effects observed especially in sore throat and overall symptom analyses （P<0.01）. Spearman correlation analysis revealed that several clinical biomarkers positively correlated with "excess heat-toxicity" syndrome and its main symptom improvement， were also called "heat-related biomarkers"， including succinic acid， α-ketoglutaric acid， glycine， lactic acid， adenosine monophosphate （AMP）， tumor necrosis factor-α （TNF-α）， interferon-γ （IFN-γ）， interleukin-1β （IL-1β）， interleukin-4 （IL-4）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， interleukin-10 （IL-10）， and so on. Conversely， clinical biomarkers negatively correlated with symptom severity， were also called "heat-clearing related biomarkers" after administration of Huanglian Jiedu Wan， including malic acid， fumaric acid， cis-aconitic acid， adrenocorticotropic hormone （ACTH）， IL-1β， IL-4， IL-8， succinic acid， and citric acid. The XGBoost classification model using all 52 biomarkers as variables achieved an average test accuracy of 0.754 and an average F1 score of 0.777. Feature importance analysis identified the scores of glutamic acid in saliva and IL-6 were the highest in all the variables， with importance scores of 0.081 and 0.080， respectively. After screening out 14 key variables and optimizing the parameters， model performance improved to an average accuracy of 0.758 and an F1 score of 0.798. Feature importance analysis further determined that the glutamic acid in saliva and IL-6 showed obvious changes after screening the variables， confirming the good syndrome prediction ability of the model constructed by these key clinical biomarkers. ConclusionThis study systematically elucidates the correlation between syndrome improvement and clinical biomarkers of Huanglian Jiedu Wan in the treatment of "excess heat-toxicity" syndrome. An XGBoost classification model based on key clinical biomarkers is successfully established， achieving effective prediction of the symptoms related to the "excess heat-toxicity" syndrome such as oral ulcers and sore throat and providing a new insight for objective identification of traditional Chinese medicine syndromes.

ObjectiveThis paper aims to find the identified and validated clinical biomarker data building upon a clinical study of early-phase phase Ⅱ and investigate the correlation analysis of Huanglian Jiedu Wan on syndrome improvement and clinical biomarkers in the treatment of "excess heat-toxicity" based on a machine learning model. Additionally， the effective prediction of clinical biomarker values for the main symptoms of the "excess heat-toxicity" syndrome was assessed. MethodsA total of 229 patients meeting the inclusion criteria for "excess heat-toxicity" syndrome were randomly divided into the Huanglian Jiedu Wan group and the placebo group. Syndrome score transition matrices were constructed for the Huanglian Jiedu Wan group and the placebo group based on three main symptoms of "excess heat-toxicity" syndrome， such as oral ulcers， sore throat， and gum swelling and pain. Data from the patients with these three syndromes were also integrated for an overall analysis. The corresponding syndrome score transition matrices were further constructed to visualize symptom change trends of the patients in the two groups via heatmaps. Based on the identified and validated clinical biomarkers related to inflammation， oxidative stress， and energy metabolism in the early phase， Spearman correlation analysis was employed to analyze and evaluate the associations between clinical biomarkers and syndrome improvement. Key clinical biomarkers reflecting the effect of Huanglian Jiedu Wan were screened through the comparison of differences between groups. An extreme gradient boosting （XGBoost） algorithm was used to develop a prediction model for main symptom classification， with classification performance evaluated through 10-fold cross-validation. Feature importance analysis was applied to identify variables with the greatest contribution to the prediction result. ResultsThe syndrome transition matrix results indicated that the Huanglian Jiedu Wan group showed a superior effect to the placebo group in improving oral ulcers， sore throat， and overall symptoms， with significant effects observed especially in sore throat and overall symptom analyses （P<0.01）. Spearman correlation analysis revealed that several clinical biomarkers positively correlated with "excess heat-toxicity" syndrome and its main symptom improvement， were also called "heat-related biomarkers"， including succinic acid， α-ketoglutaric acid， glycine， lactic acid， adenosine monophosphate （AMP）， tumor necrosis factor-α （TNF-α）， interferon-γ （IFN-γ）， interleukin-1β （IL-1β）， interleukin-4 （IL-4）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， interleukin-10 （IL-10）， and so on. Conversely， clinical biomarkers negatively correlated with symptom severity， were also called "heat-clearing related biomarkers" after administration of Huanglian Jiedu Wan， including malic acid， fumaric acid， cis-aconitic acid， adrenocorticotropic hormone （ACTH）， IL-1β， IL-4， IL-8， succinic acid， and citric acid. The XGBoost classification model using all 52 biomarkers as variables achieved an average test accuracy of 0.754 and an average F1 score of 0.777. Feature importance analysis identified the scores of glutamic acid in saliva and IL-6 were the highest in all the variables， with importance scores of 0.081 and 0.080， respectively. After screening out 14 key variables and optimizing the parameters， model performance improved to an average accuracy of 0.758 and an F1 score of 0.798. Feature importance analysis further determined that the glutamic acid in saliva and IL-6 showed obvious changes after screening the variables， confirming the good syndrome prediction ability of the model constructed by these key clinical biomarkers. ConclusionThis study systematically elucidates the correlation between syndrome improvement and clinical biomarkers of Huanglian Jiedu Wan in the treatment of "excess heat-toxicity" syndrome. An XGBoost classification model based on key clinical biomarkers is successfully established， achieving effective prediction of the symptoms related to the "excess heat-toxicity" syndrome such as oral ulcers and sore throat and providing a new insight for objective identification of traditional Chinese medicine syndromes.

Objective To prepare the recombinant Echinococcus granulosus Polo-like kinase 2 (rEgPLK2) protein and evaluate its immunoprotective efficacy against cystic echinococcosis, so as to provide insights into research and development of novel vaccines against echinococcosis. Methods The Polo-like kinase (PLK) protein sequences were retrieved from 12 species in the NCBI protein database, including E. granulosus and E. multilocularis. Multiple sequence alignment was performed using the Clustal Omega program, and structural visualization and homology analysis were conducted using the ESPript 3.2 program. The recombinant plasmid pET-30a-EgPLK2 was transformed into BL21(DE3) competent cells. Protein expression was induced with isopropyl-β-D-thiogalactoside (IPTG), and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed to characterize the expression and molecular weight of the rEgPLK2 protein. The purified rEgPLK2 protein was thoroughly emulsified with Freund’s complete adjuvant at a 1 : 1 volume ratio. Two New Zealand white rabbits were immunized with multipoint subcutaneous injection on the back at a dose of 300 μg per rabbit for primary immunization. For booster immunizations, the protein was emulsified with Freund’s incomplete adjuvant at a 1 : 1 volume ratio and administered on days 14, 28, and 42 after the primary immunization at a dose of 150 μg per rabbit. Serum was sampled from the rabbit ear vein on day 7 after the final immunization to yield anti-rEgPLK2 polyclonal antibodies. Antibody titer was determined by indirect enzyme-linked immunosorbent assay (ELISA), and antibody specificity was verified by Western blotting. The tissue localization of the EgPLK2 protein was detected in E. granulosus protoscoleces and adult worms using immunofluorescence assay (IFA). Eighteen 6- to 8-week-old female SPF-grade BALB/c mice were randomly divided into three groups, including the blank control group, rEgPLK2-ISA immunization group, and PBS-ISA adjuvant control group, of 6 mice each group. Mice in the rEgPLK2-ISA immunization group and PBSISA group received three primary immunizations via intramuscular injection, and animals in the rEgPLK2-ISA immunization group was inoculated with immunogens prepared by emulsifying rEgPLK2 protein with ISA 201 adjuvant at a 1 : 1 volume ratio (6 μg per mouse), while mice in the PBS-ISA adjuvant control group received an equal volume of PBS emulsified with ISA adjuvant at a 1 : 1 volume ratio. A fourth booster immunization was administered via intraperitoneal injection. Mice in the rEgPLK2-ISA immunization group received a booster immunization with 8 μg of rEgPLK2 protein per mouse, and animals in the PBS-ISA group received an equal volume of PBS, with immunizations given at 2-week intervals. Mice in the blank control group were given no treatment, and housed under standard conditions. Tail vein blood was collected from all mice 7 days after the final immunization, and levels of specific anti-rEgPLK2 IgG antibody and its subclasses (IgG1, IgG2a, IgG2b, IgG3) were measured by indirect ELISA. E. granulosus infection was modelled in mice through injection with 1 000 E. granulosus protoscoleces via intrahepatic portal vein in the rEgPLK2-ISA immunization group and PBS-ISA adjuvant control group 2 weeks after the last immunization. All mice were sacrificed and dissected. The number of cysts was counted in mouse livers, and the cyst reduction rate was calculated. Liver tissues were processed for paraffin sectioning and stained with hematoxylin and eosin (HE), and histopathological changes were examined under a light microscope. Results Sequence analysis revealed that EgPLK2 shared a high amino acid sequence homology with E. multilocularis PLK2 (EmPLK2) and contained the typical domains of the Polo-like kinase family, including the serine/threonine protein kinase catalytic domain (STKc) and Polo-box. The IPTG-induced rEgPLK2 protein was mainly expressed in the form of inclusion bodies, and the purified rEgPLK2 protein showed a relative molecular mass of approximately 70 kDa. The prepared rabbit anti-rEgPLK2 polyclonal antibody had a titer of 1 : 256 000, and Western blotting assay showed that this anti-body specifically recognized the rEgPLK2 protein with a relative molecular mass of approximately 70 kDa. Immunofluorescence assay showed that the EgPLK2 protein was localized in the excretory bladder and rostellum of E. granulosus protoscoleces, as well as the tegument, suckers, and inter-proglottid junctions of adult worms. Immunoprotective assay showed that the serum levels of specific anti-rEgPLK2 IgG, IgG1, IgG2a, and IgG2b antibodies were 2.92 ± 0.49, 0.33 ± 0.10, 0.31 (0.36), and 3.12 (1.73) in mice in the rEgPLK2-ISA immunization group, which were all significantly higher than those in the PBS-ISA adjuvant control group (0.14 ± 0.04, 0.07 ± 0.01, 0.12 ± 0.04, and 0.11 ± 0.04, respectively) (t = 19.28 and 8.46, Z = 3.75 and 4.15; all P values < 0.001); however, there was no significant difference in the serum anti-IgG3 antibody level between the rEgPLK2-ISA immunization group and the PBS-ISA adjuvant control group [0.07 (0.01) vs. 0.073 (0.07); Z = 0.69, P > 0.05)]. In the mouse model of E. granulosus infections, the area of hepatic lesions was reduced and the inflammatory infiltration was alleviated in the rEgPLK2-ISA immunization group than in the PBS-ISA adjuvant control group, and the number of hepatic cysts was higher in the PBS-ISA adjuvant control group than in the rEgPLK2-ISA immunization group [8.00 (2.00) vs. 1.00 (0.75); Z = −2.93, P < 0.01], with a cyst reduction rate of 80.40%. Indirect ELISA assay measured higher serum levels of specific anti-rEgPLK2 IgG (3.28 ± 0.48 vs. 0.11 ± 0.04; t = 15.86, P < 0.01), IgG1 (0.29 ± 0.02 vs. 0.09 ± 0.01; t = 15.67, P < 0.01), IgG2a [3.71 (1.09) vs. 0.08 (0.03); Z = 2.88, P < 0.01], and IgG2b antibodies [3.34 (1.01) vs. 0.08 (0.03); Z = 2.88, P < 0.01] in the rEgPLK2-ISA immunization group than in the PBS-ISA adjuvant control group, and there was no significant difference in the serum level of the specific anti-rEgPLK2 IgG3 antibody between the rEgPLK2-ISA immunization group and the PBS-ISA adjuvant control group (0.07 ± 0.01 vs. 0.07 ± 0.01; t = 1.29, P > 0.05). Conclusions The prokaryotic expression system has been successfully constructed for the EgPLK2 gene and the anti-rEgPLK2 polyclonal antibody has been obtained. The rEgPLK2 protein exhibits a high immunogenicity, and is effective to protect against E. granulosus infection, and inhibits cyst development, which is a promising candidate vaccine target against cystic echinococcosis.

Public hospitals have formed a relatively perfect working foundation in the introduction and training of young talents,but the evaluation system of young talents is not perfect.Based on the requirements of high-quality development,grasp the principle of party management of talents,combine the talents development situation in Jiading District Maternal and Child Health Care Hospital,takes the special training of young talents in administration as the starting point,comprehensively uses the literature method,interview method and Delphi method to establish the index database,uses the exploratory factor analysis meth-od to calculate the index weight,and constructs the evaluation model of young talents in hospital administrative management,so as to help hospitals better screen and evaluate talents and give full play to the value and role of talents as the first resource.

OBJECTIVE To identify tortoiseshell glue and antler glue in Qichong zuogui granules,and determine the contents of 12 chemical components.METHODS Identification and content determination were performed by using liquid chromatography-tandem mass spectrometry(LC-MS/MS)method.The identification was performed on Hypersil GOLD column with a mobile phase consisted of acetonitrile-0.1％formic acid solution(gradient elution);the electrospray ion source was used to scan in the positive ion multi-reaction detection mode.The mass charge ratio(m/z)631.3→546.4,631.3→921.4 was the detection ion pair for tortoiseshell glue,and the m/z 765.4→554.0,765.4→733.0 was the detection ion pair for antler glue.The determination method for 12 chemical components was as follows:Accucore C18 column,methanol-0.1％formic acid as mobile phase(gradient elution);scanning range of positive and negative ions was m/z 100→1 000 with the electric spray ion source and single ion detection scanning mode.RESULTS Average retention times of the molecular ion peaks for characteristic peptide segments of tortoiseshell glue and antler glue were 6.28 and 6.77 min,respectively;the linear relationship of 12 chemical components was good within their respective concentration ranges,such as astragaloside Ⅳ,calycosin-7-O-β-D-glucoside,calycosin,chlorogenic acid,ferulic acid,betaine,amygdalin,rutin,hydroxysafflor yellow A,hyperoside,loganin,cyasterone(r＞0.999);RSDs for precision,stability(24 h)and reproducibility tests were all less than 5％.The average sample recovery rates ranged from 98.04％to 101.08％.The average contents of 12 components were 1.83,25.73,13.76,56.71,23.80,49.82,807.49,15.01,317.02,60.21,202.71 and 17.70 μg/g,respectively.CONCLUSIONS In this study,tortoiseshell glue and antler glue in Qixiong zuogui granules are identified,and the contents of 12 chemical components therein are determined.This provides a reference for the quality control of this granule.

Objective Explored the clinical efficacy and mechanism of improved CO2 fractional laser on treatment for refractory wounds.Methods A total of 122 patients with refractory wounds who were admitted to the hospital from May 2022 to May 2024 were selected and randomly divided into a conventional group(conventional dressing change treatment+photodynamic therapy+conventional CO2 fractional laser treatment)and an improved group(conventional dressing change treatment+photodynamic therapy+improved CO2 fractional laser treatment)by lottery,with 61 cases in each group.The serum C-reactive protein(CRP)and erythrocyte sedimentation rate(ESR)levels,the number of wound bacteria before treatment and after 1,2,and 3 weeks of treatment were compared between the two groups.The wound healing rates after 1,2,and 3 weeks of treatment were compared between the two groups.The clinical efficacy of the two groups was compared.The levels of wound exudate basic fibroblast growth factor(bFGF)and angiopoietin-1(Ang-1)before treatment and after 1,2,and 3 weeks of treat-ment were compared.The adverse reactions of the two groups were compared.Results Repeated measures analy-sis of variance showed that there were an interaction effect between groups and over time in the serum CRP and ESR levels,the number of wound bacteria in both groups(P<0.05).There were no significant differences in the serum CRP and ESR levels and the number of wound bacteria between the two groups before treatment(P>0.05).The serum CRP and ESR levels and the number of wound bacteria in the improved group were lower than those in the conventional group after 1,2,and 3 weeks of treatment(P<0.05).Repeated measures analysis of variance showed that there was an interaction effect between groups and over time in the wound healing rates of both groups(P<0.05).The wound healing rates in the improved group were higher than those in the conventional group after 1,2,and 3 weeks of treatment(P<0.05).The total clinical effective rate of the improved group was higher than that of the conventional group(P<0.05).Repeated measures analysis of variance showed that there was an inter-action effect between groups and over time in the levels of bFGF and Ang-1 in wound exudates of both groups(P<0.05).There was no significant difference in the levels of bFGF and Ang-1 in wound exudates between the two groups before treatment(P>0.05).The levels of bFGF and Ang-1 in wound exudates in the improved group were higher than those in the conventional group after 1,2,and 3 weeks of treatment(P<0.05).There were no signifi-cant differences in the incidence of pain,pruritus and rash between the two groups(P>0.05).Conclusions In the treatment of refractory wounds,the combination of improved CO2 fractional laser and photodynamic therapy could promote the secretion of wound exudate growth factors,inhibit wound bacterial growth and inflammatory response,accelerate wound healing,and improve clinical efficacy,with high safety.The mechanism of action is related to the promotion of wound exudate growth factor secretion.

Sophora Flavescens is the dried root of the leguminous plant Sophora Flavescens Ait. It was first published in Shen Nong's Herbal Classic. Sophora Flavescens contains a variety of active ingredients, mainly including matrine and oxymatrine, with anti-inflammatory, anti-tumor, anti-arrhythmia, disease-resistant pathogenic microorganisms and other pharmacological effects. Clinically, the compound preparations of Sophora Flavescens include Compound KuShen injection and KuShen gel and so on, which can be used to treat many types of cancers and improve skin, mucous pruritus, pain and other symptoms. Due to the poor bioavailability, the structure of matrine needs to be reformed. MASM, matrine derivative, only needs a low concentration to have a good therapeutic effect on sepsis and liver fibrosis. In this article, the chemical composition, pharmacological effects, compound preparations and structural modification of matrine were mainly discussed, aiming to provide a theoretical basis for the clinical application of Sophora Flavescens and the development of new drugs.

Objective Explored the clinical efficacy and mechanism of improved CO2 fractional laser on treatment for refractory wounds.Methods A total of 122 patients with refractory wounds who were admitted to the hospital from May 2022 to May 2024 were selected and randomly divided into a conventional group(conventional dressing change treatment+photodynamic therapy+conventional CO2 fractional laser treatment)and an improved group(conventional dressing change treatment+photodynamic therapy+improved CO2 fractional laser treatment)by lottery,with 61 cases in each group.The serum C-reactive protein(CRP)and erythrocyte sedimentation rate(ESR)levels,the number of wound bacteria before treatment and after 1,2,and 3 weeks of treatment were compared between the two groups.The wound healing rates after 1,2,and 3 weeks of treatment were compared between the two groups.The clinical efficacy of the two groups was compared.The levels of wound exudate basic fibroblast growth factor(bFGF)and angiopoietin-1(Ang-1)before treatment and after 1,2,and 3 weeks of treat-ment were compared.The adverse reactions of the two groups were compared.Results Repeated measures analy-sis of variance showed that there were an interaction effect between groups and over time in the serum CRP and ESR levels,the number of wound bacteria in both groups(P<0.05).There were no significant differences in the serum CRP and ESR levels and the number of wound bacteria between the two groups before treatment(P>0.05).The serum CRP and ESR levels and the number of wound bacteria in the improved group were lower than those in the conventional group after 1,2,and 3 weeks of treatment(P<0.05).Repeated measures analysis of variance showed that there was an interaction effect between groups and over time in the wound healing rates of both groups(P<0.05).The wound healing rates in the improved group were higher than those in the conventional group after 1,2,and 3 weeks of treatment(P<0.05).The total clinical effective rate of the improved group was higher than that of the conventional group(P<0.05).Repeated measures analysis of variance showed that there was an inter-action effect between groups and over time in the levels of bFGF and Ang-1 in wound exudates of both groups(P<0.05).There was no significant difference in the levels of bFGF and Ang-1 in wound exudates between the two groups before treatment(P>0.05).The levels of bFGF and Ang-1 in wound exudates in the improved group were higher than those in the conventional group after 1,2,and 3 weeks of treatment(P<0.05).There were no signifi-cant differences in the incidence of pain,pruritus and rash between the two groups(P>0.05).Conclusions In the treatment of refractory wounds,the combination of improved CO2 fractional laser and photodynamic therapy could promote the secretion of wound exudate growth factors,inhibit wound bacterial growth and inflammatory response,accelerate wound healing,and improve clinical efficacy,with high safety.The mechanism of action is related to the promotion of wound exudate growth factor secretion.

BackgroundPostpartum depression （PPD） is a prevalent postpartum complications that significantly compromises women's psychological and physical well-being. Repetitive transcranial magnetic stimulation （rTMS）， a conventional neuromodulation technique， has been increasingly used in the treatment of PPD. However， high-quality evidence regarding its efficacy and safety remains limited. ObjectiveTo evaluate the efficacy and safety of rTMS in the treatment of PPD， thereby providing references for clinical treatment. MethodsDatabases including Cochrane Library， PubMed， Embase， CNKI， Wanfang， VIP and China Biology Medicine disc （CBM） were electronically searched for randomized controlled trials （RCTs） on rTMS for PPD， with the search spanning from database inception to February 8， 2025. Study quality was assessed using the Cochrane Handbook for Systematic Reviews of Interventions 5.0.1， and the certainty of evidence was graded according to the Grading of Recommendations Assessment， Development and Evaluation （GRADE） approach. Meta-analysis was conducted using RevMan 5.3 and Stata 12.0. The outcomes of the Meta-analysis included the total effective rate， Edinburgh Postnatal Depression Scale （EPDS） score， Hamilton Depression Rating Scale （HAMD） score， and adverse reactions （dizziness， headache， nausea， diarrhea， and the overall incidence of adverse reactions）. ResultsA total of 11 studies involving 729 patients with PPD were included. Meta-analysis results showed that the total effective rate in the study group was significantly higher than that in the control group （OR=5.54， 95% CI： 3.07–10.01， P<0.01）. Both EPDS score （SMD=-2.38， 95% CI： -3.39–-1.37， P<0.01） and HAMD score （SMD=2.53， 95% CI： 1.21–3.85， P<0.01） in the study group were significantly lower than those in the control group， with statistically significant differences. Comparisons between the study group and control group reveal no significant differences in the incidence of dizziness and headache （RR=1.47， 95% CI： 0.63–3.43， P>0.05）， nausea （RR=1.46， 95% CI： 0.55–3.86， P>0.05）， diarrhea （RR=0.71， 95% CI： 0.23–2.20， P>0.05）， and overall adverse reactions （RR=1.30， 95% CI： 0.79–2.15， P>0.05）. GRADE assessment rated the four indicators of dizziness and headache， diarrhea， overall incidence of adverse reactions， and EPDS score as "moderate-certainty evidence"， and rated the total effective rate， nausea， and the HAMD score as "low-certainty evidence". ConclusionrTMS demonstrates certain therapeutic efficacy for PPD， with a safety profile comparable to conventional treatment. ［Funded by Sichuan Psychological Society Research Planning Project （number， SCSXLXH202403099）； Guiding Science and Technology Plan Project of Guangyuan （number， 23ZDYF0095）］

Objective:To evaluate the application of metagenomic next-generation sequencing（mNGS）in the identification of non-tuberculous mycobacteria（NTM）.Methods:A retrospective analysis was conducted on mNGS results of 358 bronchoalveolar lavage fluid（BALF）samples positive for NTM collected at the First Affiliated Hospital of Zhejiang University School of Medicine from February 2021 to January 2024. The analysis included the distribution of NTM species，the detection of mixed pathogens，and the performance of conventional mycobacterial detection methods.Results:The results showed that 362 strains of 15 NTM species were identified from 350 specimens，8 specimens were not precise to the species level. The most frequently detected species were Mycobacterium intracellulare（37.3%，135/362）， Mycobacterium abscessus（26.8%，97/362），followed by Mycobacterium avium（11.0%，40/362）， Mycobacterium kansasii（8.0%，29/362）and Mycobacterium chelonae（7.7%，28/362）. Single NTM species were detected in 339 specimens，while two or three NTM species were simultaneously detected in 11 specimens（3.1%，11/358）. Non-NTM microorganisms co-infected were detected in 53.4%（191/358）of NTM-positive BALF samples，including common pathogens such as Pseudomonas aeruginosa， Staphylococcus aureus，and Aspergillus fumigatus；and difficult-to-identify pathogens such as Legionella pneumophila and Talaromyces marneffei. In NTM-positive patients detected by mNGS，the results supported the diagnosis of NTM infection in 298 cases（298/358，83.2%）and 105 cases（105/358，29.3%）initiated anti-NTM treatment accordingly；while in 60 cases（60/358，16.8%）the positive results were considered as colonization or unrelated to clinical infection. For samples tested with acid-fast staining，mycobacterial liquid culture，and DNA microarray，the positivity rates for NTM were 31.5%（73/232），48.7%（57/117），and 43.0%（46/107），respectively. Conclusions:mNGS demonstrates advantages in identification of NTM. However，the test may detect multiple microorganisms，in that case，the interpretation with clinical and radiological results is requried to determine the main pathogens.