ObjectiveHeadspace solid-phase microextraction-gas chromatography-mass spectrometry（HS-SPME-GC-MS） and GC-triple quadrupole MS（GC-QqQ-MS） in combination with non-targeted and targeted metabolomics were employed to systematically analyze the chemical composition differences of Xihuangwan prepared with natural musk and artificial musk， and establish an identification system for them. MethodsThe volatile components of 9 batches of Xihuangwan samples from 8 manufacturers were analyzed by HS-SPME-GC-MS non-targeted metabolomics， and identified by comparing their MS data with the National Institute of Standards and Technology（NIST） spectral library. Orthogonal partial least squares-discriminant analysis（OPLS-DA） was used to identify differential volatile components of Xihuangwan prepared with natural musk and artificial musk. Additionally， GC-QqQ-MS targeted metabolomics was applied to quantify the levels of α-pinene， β-elemene， muscone， dehydroepiandrosterone， bornyl acetate， and octyl acetate in 27 batches of samples from 9 manufacturers. Cluster analysis， principal component analysis（PCA）， and partial least squares-discriminant analysis（PLS-DA） were conducted to further explore the differences in volatile components between Xihuangwan samples prepared with natural musk and artificial musk. ResultsNon-targeted metabolomics identified 291 volatile compounds in Xihuangwan， including alkanes， esters， alkanes， alcohols， ketones， naphthalenes and others. OPLS-DA analysis revealed distinct separation between Xihuangwan samples containing artificial musk（A1， C1， D1， E1， F1， G1， I1） and those containing natural musk（H1， H3）. A total of 30 differential metabolites were identified. The relative contents of these 30 differential metabolites were visualized using a radar chart， revealing significant differences in the levels of octanol， borneol acetate and muscone. Cluster analysis and PCA results from targeted metabolomics indicated that Xihuangwan could be classified into two distinct groups：one composed of natural musk（H1， H3） and the other of artificial musk， sample H2. PLS-DA identified muscone， octyl acetate， and dehydroepiandrosterone as key differential volatile components. Although no significant difference was observed in the content of octyl acetate between the two groups， statistically significant differences were found for muscone and dehydroepiandrosterone（P<0.05）. ConclusionMuscone and dehydroepiandrosterone can be used for the differentiation of Xihuangwan samples containing natural musk from those containing artificial musk. This study systematically and comprehensively analyzed the differences in the types and contents of major volatile components in Xihuangwan prepared with natural musk and artificial musk， providing a scientific basis for quality evaluation and control of Xihuangwan.

ObjectiveHeadspace solid-phase microextraction-gas chromatography-mass spectrometry（HS-SPME-GC-MS） and GC-triple quadrupole MS（GC-QqQ-MS） in combination with non-targeted and targeted metabolomics were employed to systematically analyze the chemical composition differences of Xihuangwan prepared with natural musk and artificial musk， and establish an identification system for them. MethodsThe volatile components of 9 batches of Xihuangwan samples from 8 manufacturers were analyzed by HS-SPME-GC-MS non-targeted metabolomics， and identified by comparing their MS data with the National Institute of Standards and Technology（NIST） spectral library. Orthogonal partial least squares-discriminant analysis（OPLS-DA） was used to identify differential volatile components of Xihuangwan prepared with natural musk and artificial musk. Additionally， GC-QqQ-MS targeted metabolomics was applied to quantify the levels of α-pinene， β-elemene， muscone， dehydroepiandrosterone， bornyl acetate， and octyl acetate in 27 batches of samples from 9 manufacturers. Cluster analysis， principal component analysis（PCA）， and partial least squares-discriminant analysis（PLS-DA） were conducted to further explore the differences in volatile components between Xihuangwan samples prepared with natural musk and artificial musk. ResultsNon-targeted metabolomics identified 291 volatile compounds in Xihuangwan， including alkanes， esters， alkanes， alcohols， ketones， naphthalenes and others. OPLS-DA analysis revealed distinct separation between Xihuangwan samples containing artificial musk（A1， C1， D1， E1， F1， G1， I1） and those containing natural musk（H1， H3）. A total of 30 differential metabolites were identified. The relative contents of these 30 differential metabolites were visualized using a radar chart， revealing significant differences in the levels of octanol， borneol acetate and muscone. Cluster analysis and PCA results from targeted metabolomics indicated that Xihuangwan could be classified into two distinct groups：one composed of natural musk（H1， H3） and the other of artificial musk， sample H2. PLS-DA identified muscone， octyl acetate， and dehydroepiandrosterone as key differential volatile components. Although no significant difference was observed in the content of octyl acetate between the two groups， statistically significant differences were found for muscone and dehydroepiandrosterone（P<0.05）. ConclusionMuscone and dehydroepiandrosterone can be used for the differentiation of Xihuangwan samples containing natural musk from those containing artificial musk. This study systematically and comprehensively analyzed the differences in the types and contents of major volatile components in Xihuangwan prepared with natural musk and artificial musk， providing a scientific basis for quality evaluation and control of Xihuangwan.

Long-term hypertension causes excessive vascular remodeling and leads to adverse cardiovascular events. Balance of ubiquitination and deubiquitination has been linked to several chronic conditions, including pathological vascular remodeling. In this study, we discovered that the expression of ubiquitin-specific protease 25 (USP25) is significantly up-regulated in angiotensin II (Ang II)-challenged mouse aorta. Knockout of Usp25 augments Ang II-induced vascular injury such as fibrosis and endothelial to mesenchymal transition (EndMT). Mechanistically, we found that USP25 interacts directly with Forkhead box O3 (FOXO3) and removes the K63-linked ubiquitin chain on the K258 site of FOXO3. We also showed that this USP25-mediated deubiquitination of FOXO3 increases its binding to light chain 3 beta isoform and autophagosomic-lysosomal degradation of FOXO3. In addition, we further validated the biological function of USP25 by overexpressing USP25 in the mouse aorta with AAV9 vectors. Our studies identified FOXO3 as a new substrate of USP25 and showed that USP25 may be a potential therapeutic target for excessive vascular remodeling-associated diseases.

Doxorubicin (Dox) is an anthracycline drug widely applied in various malignancies. However, the fatal cardiotoxicity induced by Dox limits its clinical application. Post-transcriptional protein modification via ubiquitination/deubiquitination in cardiomyocytes mediates the pathophysiological process in Dox-induced cardiotoxicity (DIC). In this study, we aimed to clarify the regulatory role and mechanism of a deubiquitinating enzyme, ubiquitin-specific peptidase 13 (USP13), in DIC. RNA-seq analysis and experimental examinations identified that cardiomyocyte-derived USP13 positively correlated with DIC. Mice with cardiac-specific deletion of USP13 were subjected to Dox modeling. Adeno-associated virus serotype 9 (AAV9) carrying cTNT promoter was constructed to overexpress USP13 in mouse heart tissues. Cardiomyocyte-specific knockout of USP13 exacerbated DIC, while its overexpression mitigated DIC in mice. Mechanistically, USP13 deubiquitinates the stimulator of interferon genes (STING) and promotes the autolysosome-related degradation of STING, subsequently alleviating cardiomyocyte inflammation and death. Our study suggests that USP13 serves a cardioprotective role in DIC and indicates USP13 as a potential therapeutic target for DIC treatment.

With the rapid development of competitive sports, the incidence of anterior cruciate ligament (ACL) injury is on the rise. Such injuries may shorten athletes′ career and lead to other long-term adverse consequences. Although athletes generally recover well after ACL reconstruction, many still struggle to return to their pre-injury performance levels. Advances in the understanding of ACL anatomy and injury mechanisms, along with the evolution of surgical techniques and rehabilitation methods, have provided more individualized and tailored options for athletes following ACL injuries. However, there is currently no consensus in China regarding surgical and rehabilitation strategies for competitive athletes aiming to return to sports after ACL injuries. To this end, the Sports Medicine Committee of the Chinese Research Hospital Association and the Editorial Board of the Chinese Journal of Trauma jointly formulated the Expert consensus on surgical treatment and rehabilitation for competitive sports athletes returning to sports after anterior cruciate ligament injury ( version 2025), and presented 14 recommendations covering surgical indications, preoperative rehabilitation, surgical timing, surgical strategies and postoperative rehabilitation strategies, aiming to improve the surgical treatment and rehabilitation system for ACL injuries in competitive athletes and facilitate their return to high-level sports performance after injury.

Objective:Investigate the clinical characteristics of patients with coronary artery disease (CAD) and moderate-to-severe coronary calcification lesions confirmed by intravascular ultrasound (IVUS), and to compare the prevalence of moderate-to-severe calcification lesions among different clusters of patients with CAD.Methods:This cross-sectional study enrolled patients with coronary artery disease who underwent coronary angiography and IVUS at the General Hospital of Northern Theater Command from June 2016 to June 2023. Patients with mild calcification were excluded, and the remaining cohort was divided into moderate-to-severe calcification and non-calcification groups. The least absolute and selective operator logistic regression was used to identify baseline variables associated with mederate-to-severe calcification. Hierarchical cluster analysis was used to identify similarities in clinical baseline data among patients, and the prevalence of moderate-to-severe coronary calcification was compared across different clusters of patients with CAD.Results:A total of 1 406 patients with CAD were enrolled (age (60.7±10.0) years; 963 males (68.5%)). A total of 563 patients were assigned in the moderate-to-severe calcification group and 843 patients were assigned in the non-calcification group. The least absolute and selective operator logistic regression identified 23 baseline variables associated with moderate-to-severe coronary artery calcification. Hierarchical cluster analysis based on these 23 variables divided all patients into Cluster 1 (500 cases) and Cluster 2 (906 cases). Patients in Cluster 1 were characterized by high age, high body mass index, high diastolic blood pressure, elevated mean arterial pressure, history of hypertension, history of diabetes, low endogenous creatinine clearance, high fasting blood glucose, low high-density lipoprotein cholesterol (HDL-C), high lipoprotein(a), high alkaline phosphatase, high glycated hemoglobin, low serum albumin, high triglycerides/HDL-C ratio, high HbA1c/HDL-C ratio, and high triglyceride glucose index. Cluster 1 (49.6%, 248/500) exhibited a significantly higher prevalence of moderate-to-severe coronary artery calcification compared to Cluster 2 (34.8%, 315/906, P=0.002). Conclusions:Patients with moderate-to-severe coronary artery calcification exhibited clinical heterogeneity compared to those without calcification. A subgroup characterized by high age, elevated body mass index, high diastolic blood pressure, elevated mean arterial pressure, history of hypertension, history of diabetes, low endogenous creatinine clearance, and high fasting blood glucose levels demonstrated a higher prevalence of moderate-to-severe coronary artery calcification.

Objective:To establish a quality standard for the bacterial endotoxin test method of fructose sodium diphosphate injections(FDP injections),so as to provide reference for the formulation and revision of the national standard for this drug.Methods:The interference test and bacterial endotoxins test of 22 batch samples of FDP in-jections from nine manufacturers were performed with TALs,and the bacterial endotoxins of samples were tested and the results were judged.Results:A bacterial endotoxin limit value of 0.72 EU·mg-1 for fructose sodium diphos-phate was established,which was suitable for 22 batches of FDP injections met the requirements.Conclusion:The bacterial endotoxin test method established in this study can be used to substitute the rabbit pyrogen test and meet the quality control requirements of FDP injections.

Objective:To establish a quality standard for the bacterial endotoxin test method of fructose sodium diphosphate injections(FDP injections),so as to provide reference for the formulation and revision of the national standard for this drug.Methods:The interference test and bacterial endotoxins test of 22 batch samples of FDP in-jections from nine manufacturers were performed with TALs,and the bacterial endotoxins of samples were tested and the results were judged.Results:A bacterial endotoxin limit value of 0.72 EU·mg-1 for fructose sodium diphos-phate was established,which was suitable for 22 batches of FDP injections met the requirements.Conclusion:The bacterial endotoxin test method established in this study can be used to substitute the rabbit pyrogen test and meet the quality control requirements of FDP injections.

With the rapid development of competitive sports, the incidence of anterior cruciate ligament (ACL) injury is on the rise. Such injuries may shorten athletes′ career and lead to other long-term adverse consequences. Although athletes generally recover well after ACL reconstruction, many still struggle to return to their pre-injury performance levels. Advances in the understanding of ACL anatomy and injury mechanisms, along with the evolution of surgical techniques and rehabilitation methods, have provided more individualized and tailored options for athletes following ACL injuries. However, there is currently no consensus in China regarding surgical and rehabilitation strategies for competitive athletes aiming to return to sports after ACL injuries. To this end, the Sports Medicine Committee of the Chinese Research Hospital Association and the Editorial Board of the Chinese Journal of Trauma jointly formulated the Expert consensus on surgical treatment and rehabilitation for competitive sports athletes returning to sports after anterior cruciate ligament injury ( version 2025), and presented 14 recommendations covering surgical indications, preoperative rehabilitation, surgical timing, surgical strategies and postoperative rehabilitation strategies, aiming to improve the surgical treatment and rehabilitation system for ACL injuries in competitive athletes and facilitate their return to high-level sports performance after injury.

Objective:Investigate the clinical characteristics of patients with coronary artery disease (CAD) and moderate-to-severe coronary calcification lesions confirmed by intravascular ultrasound (IVUS), and to compare the prevalence of moderate-to-severe calcification lesions among different clusters of patients with CAD.Methods:This cross-sectional study enrolled patients with coronary artery disease who underwent coronary angiography and IVUS at the General Hospital of Northern Theater Command from June 2016 to June 2023. Patients with mild calcification were excluded, and the remaining cohort was divided into moderate-to-severe calcification and non-calcification groups. The least absolute and selective operator logistic regression was used to identify baseline variables associated with mederate-to-severe calcification. Hierarchical cluster analysis was used to identify similarities in clinical baseline data among patients, and the prevalence of moderate-to-severe coronary calcification was compared across different clusters of patients with CAD.Results:A total of 1 406 patients with CAD were enrolled (age (60.7±10.0) years; 963 males (68.5%)). A total of 563 patients were assigned in the moderate-to-severe calcification group and 843 patients were assigned in the non-calcification group. The least absolute and selective operator logistic regression identified 23 baseline variables associated with moderate-to-severe coronary artery calcification. Hierarchical cluster analysis based on these 23 variables divided all patients into Cluster 1 (500 cases) and Cluster 2 (906 cases). Patients in Cluster 1 were characterized by high age, high body mass index, high diastolic blood pressure, elevated mean arterial pressure, history of hypertension, history of diabetes, low endogenous creatinine clearance, high fasting blood glucose, low high-density lipoprotein cholesterol (HDL-C), high lipoprotein(a), high alkaline phosphatase, high glycated hemoglobin, low serum albumin, high triglycerides/HDL-C ratio, high HbA1c/HDL-C ratio, and high triglyceride glucose index. Cluster 1 (49.6%, 248/500) exhibited a significantly higher prevalence of moderate-to-severe coronary artery calcification compared to Cluster 2 (34.8%, 315/906, P=0.002). Conclusions:Patients with moderate-to-severe coronary artery calcification exhibited clinical heterogeneity compared to those without calcification. A subgroup characterized by high age, elevated body mass index, high diastolic blood pressure, elevated mean arterial pressure, history of hypertension, history of diabetes, low endogenous creatinine clearance, and high fasting blood glucose levels demonstrated a higher prevalence of moderate-to-severe coronary artery calcification.