Background: Inhalation-based combination therapy has gained increasing attention for local treatments. However, a key challenge remains in ensuring the sustained pulmonary release of multiple active ingredients, particularly in traditional Chinese medicine (TCM) formulations. Objective: This study investigates a novel PulmoSphere-based inhalable carrier designed for the sustained pulmonary release of multiple active ingredients, using Shuang-Huang-Lian as a model TCM formulation containing three chemical markers. Materials and methods: The carrier was developed using PulmoSphere technology, incorporating phospholipid complexes of the chemical markers and hyaluronic acid (HA) into spray-dried microparticles. The aerodynamic properties, release characteristics, pulmonary distribution, and anti-inflammatory efficacy of different formulations were evaluated in vitro and in mice. Results: The microparticles retained the excellent aerodynamic properties of conventional PulmoSphere particles, with a mass median aerodynamic diameter of approximately 3.1 μm and a fine particle fraction of approximately 55%. Compared to free Shuang-Huang-Lian or phospholipid complex-loaded PulmoSphere particles, the HA-containing particles prolonged the retention of chemical markers in the lung epithelial lining fluid, demonstrating sustained release in vivo. Additionally, the HA-containing formulation enhanced the exposure of the three chemical markers to immune cells and lung tissues, leading to improved and prolonged anti-inflammatory effects, even at decreased doses. Conclusion: This novel inhalable particle system represents a promising approach for sustained pulmonary co-delivery of multiple active ingredients, offering enhanced and extended local therapeutic efficacy.

Objective To comparatively evaluate the diagnostic value of metagenomic next-generation sequencing(mNGS)versus conventional microbial culture in spinal infections.Methods A cross-section design was conducted on 82 consecutive patients with suspected spinal infections treated between February 2022 and January 2024 at Jiangbei Branch of First Affiliated Hospital of Army Medical University(Third Military Medical University).Microbiological culture,histopathological examination,and mNGS results from infected specimens were analyzed.Clinical diagnosis,primarily based on clinical manifestations,laboratory tests and radiologic features combined with medical history,was defined as the gold standard,and then the diagnostic performance,including sensitivity and specificity,were compared between mNGS and microbial culture.Results Among the 82 patients,definitive microbiological evidence was identified in 70 cases,and mNGS demonstrated a significantly higher detection rate than microbial culture(64 vs 36 cases,78.05％vs 43.9％,P<0.05).mNGS also obtained obviously higher sensitivity,accuracy,and negative predictive value(NPV),and notably lower positive predictive value(PPV)when compared to conventional microbial culture(all P<0.05).When stratified by infection type,mNGS obtained significantly higher sensitivity and accuracy compared to microbial culture in tuberculous spinal infections(P<0.05).For non-tuberculous spinal infections,mNGS also showed superior sensitivity to microbial culture(P<0.05).Conclusion In patients with spinal infections,mNGS demonstrates a significantly higher pathogen detection rate than conventional microbial culture.This technique can provide early and broad-spectrum pathogenic microbiological evidence for spinal infection.

Diabetic cardiomyopathy is a distinct form of cardiomyopathy that can lead to heart failure, arrhythmias, cardiogenic shock, and sudden death. It has become a major cause of mortality in diabetic patients. The pathogenesis of diabetic cardiomyopathy is complex, involving increased oxidative stress, activation of inflammatory responses, disturbances in glucose and lipid metabolism, accumulation of advanced glycation end products (AGEs), abnormal autophagy and apoptosis, insulin resistance, and impaired intracellular Ca²⁺ homeostasis. Recent studies have shown that adenosine monophosphate-activated protein kinase (AMPK) plays a crucial protective role by lowering blood glucose levels, promoting lipolysis, inhibiting lipid synthesis, and exerting antioxidant, anti-inflammatory, anti-apoptotic, and anti-ferroptotic effects. It also enhances autophagy, thereby alleviating myocardial injury under hyperglycemic conditions. Consequently, AMPK is considered a key protective factor in diabetic cardiomyopathy. As part of diabetes prevention and treatment strategies, both pharmacological and exercise interventions have been shown to mitigate diabetic cardiomyopathy by modulating the AMPK signaling pathway. However, the precise regulatory mechanisms, optimal intervention strategies, and clinical translation require further investigation. This review summarizes the role of AMPK in the prevention and treatment of diabetic cardiomyopathy through drug and/or exercise interventions, aiming to provide a reference for the development and application of AMPK-targeted therapies. First, several classical AMPK activators (e.g., AICAR, A-769662, O-304, and metformin) have been shown to enhance autophagy and glucose uptake while inhibiting oxidative stress and inflammatory responses by increasing the phosphorylation of AMPK and its downstream target, mammalian target of rapamycin (mTOR), and/or by upregulating the gene expression of glucose transporters GLUT1 and GLUT4. Second, many antidiabetic agents (e.g., teneligliptin, liraglutide, exenatide, semaglutide, canagliflozin, dapagliflozin, and empagliflozin) can promote autophagy, reverse excessive apoptosis and autophagy, and alleviate oxidative stress and inflammation by enhancing AMPK phosphorylation and its downstream targets, such as mTOR, or by increasing the expression of silent information regulator 1 (SIRT1) and peroxisome proliferator-activated receptor‑α (PPAR‑α). Third, certain anti-anginal (e.g., trimetazidine, nicorandil), anti-asthmatic (e.g., farrerol), antibacterial (e.g., sodium houttuyfonate), and antibiotic (e.g., minocycline) agents have been shown to promote autophagy/mitophagy, mitochondrial biogenesis, and inhibit oxidative stress and lipid accumulation via AMPK phosphorylation and its downstream targets such as protein kinase B (PKB/AKT) and/or PPAR‑α. Fourth, natural compounds (e.g., dihydromyricetin, quercetin, resveratrol, berberine, platycodin D, asiaticoside, cinnamaldehyde, and icariin) can upregulate AMPK phosphorylation and downstream targets such as AKT, mTOR, and/or the expression of nuclear factor erythroid 2-related factor 2 (Nrf2), thereby exerting anti-inflammatory, anti-apoptotic, anti-pyroptotic, antioxidant, and pro-autophagic effects. Fifth, moderate exercise (e.g., continuous or intermittent aerobic exercise, aerobic combined with resistance training, or high-intensity interval training) can activate AMPK and its downstream targets (e.g., acetyl-CoA carboxylase (ACC), GLUT4, PPARγ coactivator-1α (PGC-1α), PPAR-α, and forkhead box protein O3 (FOXO3)) to promote fatty acid oxidation and glucose uptake, and to inhibit oxidative stress and excessive mitochondrial fission. Finally, the combination of liraglutide and aerobic interval training has been shown to activate the AMPK/FOXO1 pathway, thereby reducing excessive myocardial fatty acid uptake and oxidation. This combination therapy offers superior improvement in cardiac dysfunction, myocardial hypertrophy, and fibrosis in diabetic conditions compared to liraglutide or exercise alone.

OBJECTIVES: To investigate the expression of parathyroid hormone-like hormone (PTHLH) in hepatocellular carcinoma (HCC) and analyze its correlation with clinical prognosis, its regulatory effects on HCC cell behaviors, and the signaling pathways mediating its effects. METHODS: We analyzed the differential expression of PTHLH in HCC and adjacent tissues and its association with patient prognosis based on data from TCGA and GEO databases and from 70 HCC patients treated in our hospital. The effects of PTHLH knockdown and overexpression on proliferation, migration, and invasion of cultured HCC cells were investigated using CCK-8 assay, colony formation assay, Transwell migration and invasion assays, and the signaling pathways activated by PTHLH were detected using Western blotting. RESULTS: TCGA and GEO database analysis showed significant overexpression of PTHLH mRNA in HCC tissues, which was associated with poor prognosis of the patients (P<0.05). High PTHLH mRNA expression was a probable independent prognostic risk factor for HCC (P<0.05). In the clinical samples, PTHLH mRNA and protein expressions were significantly higher in HCC tissues than in the adjacent tissues (P<0.001 or 0.01). Univariate and multivariate Cox regression analyses suggested that high PTHLH mRNA expression was an independent risk factor to affect postoperative disease-free survival of HCC patients (P<0.05). The prognostic prediction model based on PTHLH mRNA expression showed an improved accuracy for predicting the risk of postoperative recurrence in HCC patients. In cultured HCC cells, PTHLH overexpression significantly promoted cell proliferation, colony formation, migration and invasion, and caused activation of the ERK/JNK signaling pathway in Huh7 and Hep3B cells. CONCLUSIONS High PTHLH expression promotes HCC progression and is associated with poor patient prognosis. Its pro-tumor effects may be mediated by activation of the ERK/JNK signaling pathway.
Humans ; Carcinoma, Hepatocellular/metabolism* ; Liver Neoplasms/metabolism* ; Prognosis ; Cell Proliferation ; Parathyroid Hormone-Related Protein/genetics* ; Cell Line, Tumor ; Cell Movement ; Disease Progression ; Signal Transduction ; Male ; RNA, Messenger/genetics* ; Female

This study aims to investigate the ameliorating effect of Corni Fructus(CF) on the myocardial ischemic injury and the pharmacokinetic properties of characteristic components of CF. The mouse model of isoproterenol-induced myocardial ischemia was established and administrated with the aqueous extract of CF. The general efficacy of CF in ameliorating the myocardial ischemic injury was evaluated based on the cardiac histopathology and the levels of myocardial injury markers: creatine kinase isoenzyme(CK-MB) and cardiac troponin I(cTn-I). The metabolomics analysis was carried out for the heart and serum samples of mice to screen the biomarkers of CF in ameliorating the myocardial ischemic injury and then the predicted biomarkers were submitted to metabolic pathway enrichment. The pharmacokinetic analysis was performed for morroniside, loganin, and cornuside Ⅰ in mouse heart and serum samples to obtain the pharmacokinetic parameters of these components. The pharmacokinetic parameters were then integrated on the basis of self-defined weighting coefficients to simulate an integrated pharmacokinetic profile of CF iridoid glycosides in the heart and serum of the mouse model of myocardial ischemia. The results indicated that CF reduced the pathological damage to cardiac cells and tissue(hematoxylin-eosin staining) and lowered the levels of CK-MB and cTn-I in the serum of the mouse model of myocardial ischemia(P<0.01). Metabolomics analysis screed out 31 endogenous metabolites in the heart and 35 in the serum as biomarkers of CF in ameliorating the myocardial ischemic injury. These biomarkers were altered by modeling and restored by CF. Six metabolic pathways in the heart and 5 in the serum were enriched based on these metabolic markers. The main integrated pharmacokinetic parameters of CF iridoid glycosides were T_(max)=1 h, t_(1/2)=(1.52±0.05) h in the heart and T_(max)=1 h, t_(1/2)=(1.56±0.50) h in the serum. Both concentration-time curves showed a double-peak phenomenon. In conclusion, CF demonstrated the cardioprotective effect by regulating metabolic pathways such as taurine and hypotaurine metabolism, and pantothenic acid and coenzyme A biosynthesis. The integrated pharmacokinetics reflect the general pharmacokinetic properties of characteristic components in CF.
Animals ; Cornus/chemistry* ; Mice ; Metabolomics ; Drugs, Chinese Herbal/administration & dosage* ; Male ; Myocardial Ischemia/metabolism* ; Humans ; Troponin I/metabolism* ; Myocardium/pathology* ; Disease Models, Animal ; Biomarkers/metabolism* ; Creatine Kinase, MB Form/metabolism*

Objective To observe the clinical efficacy of successive trigger needling combined with fire needling in the treatment of rheumatoid arthritis of cold-damp obstruction type.Methods A total of 72 patients with rheumatoid arthritis of cold-damp obstruction type were randomly divided into observation group and control group,36 cases in each group.The control group was treated with fire needling combined with ordinary acupuncture,and the observation group was treated with fire needling combined with filiform needle.The treatments were performed once every other day,3 times a week,2 weeks as a course of treatment,continuous treatment for 2 courses.After 4 weeks of treatment,the clinical efficacy of the two groups was evaluated.The changes of pain Visual Analogue Scale(VAS)score and Self-Rating Anxiety Scale(SAS)score before and after treatment,and the time of morning stiffness of the joints were observed in the two groups.The changes of the number of joint swelling and the number of joint tenderness were compared before and after treatment between the two groups.Results(1)After treatment,the time of morning stiffness of the joints of the two groups of patients was significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving morning stiffness of the joints,and the difference was statistically significant(P＜0.05).(2)After treatment,the number of swollen joints and the number of joint pressure and pain in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving swollen joints and joint pressure and pain,and the difference was statistically significant(P＜0.05).(3)After treatment,the VAS and SAS scores of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving the VAS and SAS scores,and the difference was statistically significant(P＜0.05).(4)The total effective rate of the observation group was 91.66%(33/36),while that of the control group was 77.78%(28/36).The efficacy of the observation group was superior to that of the control group,and the difference was statistically significant(P＜0.05).Conclusion Successive trigger needling combined with fire needling treating rheumatoid arthritis of cold-damp obstruction type can significantly improve the clinical symptoms of patients,reduce their anxiety,and thus improve the quality of life of patients,with remarkable efficacy.

Spinal surgical site infection (SSI), especially deep SSI after internal fixation is difficult in treatment, with long course of disease and poor prognosis. At present, there are many controversies in the diagnosis and treatment of spinal SSI, with unsatisfactory overall efficacy of its diagnosis and treatment. Besides, no diagnosis and treatment guideline based on evidence-based medicine has been in existence. To this end, the Spinal Infection Group of the Orthopedic Branch of the Chinese Medical Doctor Association and the Spinal Infection Group of the Spinal Surgery Branch of the Chinese Rehabilitation Medicine Association jointly organized relevant experts to formulate Evidence-based clinical guideline for the diagnosis and treatment of surgical site infection in spinal trauma ( version 2024) based on an evidence-based approach. A total of 10 recommendations were proposed on the diagnosis and treatment of spinal SSI, so as to provide a clinical reference for the diagnosis and treatment of spinal SSI.

Objective:To detect the expression of autophagy-associated genes (ATGs) involved in the early stage of autophagy in peripheral blood mononuclear cells (PBMCs) of patients with ankylosing spondylitis (AS) and analyze the results with clinical data. To explore the association between the early stage of autophagy and AS and the clinical significance of ATGs involved in the early stage of autophagy in assessing the disease and inflammatory status of AS.Methods:①Clinical data and peripheral blood samples of 90 patients with AS (AS group) who were admitted to the Rheumatology and Immunology Department of the Affiliated Hospital of North Sichuan Medical College from March 2021 to August 2022 were collected, including 32 patients in the active stage (ASA group) and 58 patients in the stable stage (ASS group). In addition, clinical data and peripheral blood samples of 30 patients who were treated with secuchiumab for 24 weeks were also collected. The clinical data and peripheral blood samples of 45 healthy control (HC group) who underwent physical examination at the Affiliated Hospital of Sichuan Northwest Medical University at the same time served as controls for clinical data and peripheral blood specimens were used as controls. RT-qPCR was used to detect the mRNA expression levels of 8 ATGs (ATG13, ATG14, ATG17, ATG18, ATG101, Beclin1, ULK1, mTOR) involved in the early stage of autophagy in all PBMCs of peripheral blood samples, and compared between different groups. Data that follows a normal distribution is tested using the t-test, while data that does not follow a normal distribution is tested using the Wilcoxon rank sum test. Correlation analysis is performed using Spearman's rank correlation coefficient.②Receiver operating curve (ROC) was used to evaluate the value of ATGs with differential expression between AS group and HC group in detecting AS disease.Results:①Compared with HC group, the level of ATG13, ATG14, ATG17, ATG18, ATG101 and Beclin1 mRNA in AS group were significantly lower than those in HC group[ATG3:3.52(1.95, 5.09)×10 -3, 7.21(5.49, 9.16)×10 -3, Z=-5.64, P<0.001; ATG14:2.48(1.85, 3.64)×10 -3, 6.16(4.27, 7.80)×10 -3, Z=-6.44, P<0.001; ATG17: 6.45(3.29, 9.48)×10 -3, 18.52(12.30, 22.51)×10 -3, Z=-6.18, P<0.001;ATG18:2.97(1.77, 4.37)×10 -3, 4.61(3.27, 5.59)×10 -3, Z=-3.88, P<0.001; ATG101: 2.07(1.11, 3.33)×10 -3, 3.65(2.41, 5.20)×10 -3, Z=-3.87, P<0.001; Beclin1: 3.50(0.63, 6.14)×10 -3, 4.17(2.82, 7.93)×10 -3, Z=-1.82, P=0.027]. ②Comparison between ASA and ASS groups: The levels of ATG17, ATG 101 and Beclin1 mRNA in ASA group were significantly lower than those in ASS group[ATG17: 4.61(2.75, 7.85)×10 -3, 6.86(3.85, 11.28)×10 -3, Z=-2.16, P=0.030; ATG101:0.93(0.40, 1.67)×10 -3, 2.15(1.17, 3.20)×10 -3, Z=-3.94, P=0.002; Beclin1: 1.24(0.52, 3.94)×10 -3, 3.86(1.55, 5.45)×10 -3, Z=-2.26, P=0.024]. ③Spearman correlation analysis showed that the mRNA expression levels of ATG13 and Beclin1 in AS were negatively correlated with ESR and hs-CRP, respectively ( r=-0.22, P=0.038; r=-0.30, P=0.006; r=-0.34, P=0.004; r=-0.241, P=0.037), ATG18 mRNA expression ESR was positively correlated ( r=0.22, P=0.041). ④ROC curve showed that ATG13, ATG14, and ATG17 had a good ability to predict AS, and their area under the curve (AUC) was 0.821, 0.866, and 0.851, respectively. The mRNA expression levels of ATG13, ATG14, and ATG18 in AS were significantly increased after 24 weeks of secuchiumab treatment[ATG13: 3.09(0.17, 4.48)×10 -3, 3.50(3.42, 3.57)×10 -3, Z=-3.45, P=0.001; ATG14: 2.49(1.43, 4.03)×10 -3, 5.62(2.28, 6.77)×10 -3, Z=-3.01, P=0.003; ATG18: 2.91(1.61, 4.37)×10 -3, 4.53(2.91, 5.73)×10 -3, Z=-3.34, P=0.001]. Conclusion:①The different expressions of ATG13, ATG14, ATG17, ATG18, ATG101, and Beclin1 between HC and AS suggest that these 6 genes may related to the pathogenesis of AS, among which Beclin1 and ATG13 are closely related to the levels of inflammatory indicators ESR and hs-CRP in patients. It may affect the inflammatory state in AS. ②IL-17Ai may be a potential regulator of autophagy, which can play a therapeutic role by affecting the early autophagy process in AS, but the specific mechanism needs to be further explored. ③ Genes related to the early stage of autophagy are expected to be biological indicators for monitoring the occurrence of AS.

Abstract: Objective　To find out the causes and risk factors of this food-borne disease outbreak that occurred in November 2019 at four schools in Qinzhou, Guangxi, implement effective preventive and control measures to curb the spread and escalation of the outbreak, and provide a basis for the investigation and management of similar incidents in the future. Methods　Descriptive epidemiological methods were used to analyze the cases' clinical characteristics, three-dimensional distribution, and related risk factors. The etiological food was determined by the method of analytical epidemiology. A food hygiene investigation was conducted to trace the process of food contamination, and biological samples from cases and workers, suspicious food, and environmental samples were collected for laboratory testing. The detected Salmonella strains were analyzed for homogeneity using pulsed-field gel electrophoresis (PFGE). Results　The event involved four schools, with 69 suspected cases identified, resulting inan attack rate of 0.52% (69/13 307). The main clinical symptoms were abdominal pain, diarrhea, and fever, with an average incubation period of 20 hours.The epidemic curve indicated a point-source outbreak. Descriptive epidemiological methods inferred that bakery products distributed by Company F were the suspect foods. The case-control study indicated the risk of illness was increased among those who purchased and consumed food supplied by Company F (OR=37.67, 95%CI:14.03-101.13),particularly those who consumed the delicious hamburger (OR=30.13，95%CI:13.22-68.67). Salmonella Enteritidis (SE) was detected in 12 anal swabsfrom patients, one patient's vomit, one flour sample used for bread-making, and one egg,with homogeneity of 100% achieved in PFGE molecular typing of the 15 positive Salmonella strains. Conclusions　The outbreak is a food-borne disease outbreak caused by Salmonella contamination. It is recommended that regulatory authorities strengthen supervision and management of production and processing enterprises, standardize food processing processes, and establish and improve food safety management systems and long-term mechanisms for food safety knowledge promotion and education in schools to enhance students' hygiene awarenessand prevent similar incidents from occurring.

Aim To investigate the effects of bone-forming protein BMP9 on aerobic glycolysis,migration and invasion ability in triple-negative breast cancer MDA-MB-231 cells and the underlying mechanisms.Methods The experimental group infected MDA-MB-231 cells with human BMP9 recombinant adenovirus(AdBMP9),while the control group infected cells with empty GFP adenovirus.Lactate,glucose and ATP as-say kits were used to detect glucose uptake,lactate and ATP production.The correlation between BMP9 and key glycolytic enzyme genes in pancarcinoma was ana-lyzed using GEPIA2 database.The mRNA expression levels of GLUT1,HK2,PKM2 and LDHA in MDA-MB-231 cells after overexpression of BMP9 were detec-ted by qRT-PCR.Potential targets of BMP9 inhibiting MDA-MB-231 aerobic glycolysis were analyzed in STRING database.The expression levels of HIF-1αand downstream protein were detected by Western blot.The changes of cell migration and invasion ability after different treatments were evaluated by the scratch heal-ing assay and Transwell assay.Results Compared with the control group,BMP9 down-regulated glucose uptake,lactate production,ATP level(P＜0.01),and inhibited HIF-1α and its downstream protein ex-pression in MDA-MB-231 cells.Overexpression of HIF-1α in rescue experiment reversed the inhibitory effect of BMP9 on aerobic glycolysis,migration and in-vasion of MDA-MB-231 breast cancer cells.Conclu-sion BMP9 down-regulates HIF-1α to inhibit the aer-obic glycolysis and migration and invasion ability of MDA-MB-231 breast cancer cells.