An assay was established for detection of toxigenic Clostridioides difficile by combining microfluidic chip analysis with immunochromatography,and its performance was evaluated and compared with those of the Xpert C.difficile/Epi and VIDAS CD AB tests.Primer pairs were designed according to the tcdB and tpi genes in C.difficile.The specificity,limit of detection,reproducibility,and stability were evaluated.A total of 215 stool samples from patients with diarrhea were collected and tested in parallel with the Xpert C.difficile/Epi,VIDAS CDAB,and our assay.C.difficile was isolated from samples,and the tcdB gene was identified when discrepant results were obtained from the three above assays.Our assay showed no cross-reaction with other diarrhea-associated pathogens.Its reproducibility was 100％in testing of two standard plasmids containing tcdB and tpi genes at two concentrations(105 and 102 copies/μL).Two standard plasmids were detected after the PCR and immunochromatography reagents had been stored for 3,6,9,and 12 months,and all the results were posi-tive.The limit of detection was 10 copies/μL for toxigenic C.difficile.Testing of 33 samples positive for C.difficile with our assay(33/215,15.3％)yielded findings statistically coherent with those of the Xpert C.difficile/Epi test(kappa value=0.965).The sensitivity,specificity,positive predictive value,and negative predictive value of our assay,with respect to Xpert C.difficile/Epi as the standard,were 94.3％,100.0％,100.0％,and 98.9％;these values were significantly higher than those of VIDAS CDAB(60.0％,98.9％,91.3％,and 92.7％)(Kappa=0.714,OR=157.50,95％CI:62.03-847.28,P=0.013).In conclusion,our newly developed assay is specific,stable,and reproducible,and may be used for rapid and accu-rate detection of toxigenic C.difficile.The assay could be used for C.difficile infection screening in outpatient and emergen-cy,community medical service center,and epidemiological settings.

The chemical constituents from Cephalotaxus lanceolata were isolated and purified by using multiple chromatographic techniques, including octadecylsilane(ODS), silica gel, Sephadex LH-20 column chromatography, and semi-preparative high-performance liquid chromatography(HPLC). A total of 17 compounds obtained were identified by using spectroscopic methods such as nuclear magnetic resonance(NMR), mass spectrometry(MS), and ultraviolet(UV) combined with literature data. Compound 1 was a new alkaloid dimer, named cephalancetine E. The known compounds were determined as cephalancetine A(2), 11-hydroxycephalotaxine(3), 4-hydroxycephalotaxine(4), cephalotaxine(5), epicephalotaxine(6), cephalotaxine β-N-oxide(7), acetylcephalotaxine(8), cephalotine A(9), cephalotine B(10), 11-hydroxycephalotaxine hemiketal(11), 3-deoxy-3,11-epoxy-cephalotaxine(12), cephalotaxinone(13), isocephalotaxinone(14), 2,11-epoxy-1,2-dihydro-8-oxo-cephalotaxine(15), cephalotaxamide(16), and drupacine(17), respectively. Compounds 11, 12, and 15 were isolated from the Cephalotaxus genus for the first time. The biological activity was tested for compounds 1-17. The results reveal that compound 17 displays potent inhibitory activities against three human cancer cell lines(HepG-2, MCF-7, and SH-SY5Y).
Cephalotaxus/chemistry* ; Humans ; Cell Line, Tumor ; Drugs, Chinese Herbal/pharmacology* ; Harringtonines/pharmacology* ; Molecular Structure ; Dimerization ; Alkaloids/isolation & purification* ; Magnetic Resonance Spectroscopy

OBJECTIVE: Gastric cancer (GC) is one of the most common malignancies seen in clinic and requires novel treatment options. Morin is a natural flavonoid extracted from the flower stalk of a highly valuable medicinal plant Prunella vulgaris L., which exhibits an anti-cancer effect in multiple types of tumors. However, the therapeutic effect and underlying mechanism of morin in treating GC remains elusive. The study aims to explore the therapeutic effect and underlying molecular mechanisms of morin in GC. METHODS: For in vitro experiments, the proliferation inhibition of morin was measured by cell counting kit-8 assay and colony formation assay in human GC cell line MKN45, human gastric adenocarcinoma cell line AGS, and human gastric epithelial cell line GES-1; for apoptosis analysis, microscopic photography, Western blotting, ubiquitination analysis, quantitative polymerase chain reaction analysis, flow cytometry, and RNA interference technology were employed. For in vivo studies, immunohistochemistry, biomedical analysis, and Western blotting were used to assess the efficacy and safety of morin in a xenograft mouse model of GC. RESULTS: Morin significantly inhibited the proliferation of GC cells MKN45 and AGS in a dose- and time-dependent manner, but did not inhibit human gastric epithelial cells GES-1. Only the caspase inhibitor Z-VAD-FMK was able to significantly reverse the inhibition of proliferation by morin in both GC cells, suggesting that apoptosis was the main type of cell death during the treatment. Morin induced intrinsic apoptosis in a dose-dependent manner in GC cells, which mainly relied on B cell leukemia/lymphoma 2 (BCL-2) associated agonist of cell death (BAD) but not phorbol-12-myristate-13-acetate-induced protein 1. The upregulation of BAD by morin was due to blocking the ubiquitination degradation of BAD, rather than the transcription regulation and the phosphorylation of BAD. Furthermore, the combination of morin and BCL-2 inhibitor navitoclax (also known as ABT-737) produced a synergistic inhibitory effect in GC cells through amplifying apoptotic signals. In addition, morin treatment significantly suppressed the growth of GC in vivo by upregulating BAD and the subsequent activation of its downstream apoptosis pathway. CONCLUSION Morin suppressed GC by inducing apoptosis, which was mainly due to blocking the ubiquitination-based degradation of the pro-apoptotic protein BAD. The combination of morin and the BCL-2 inhibitor ABT-737 synergistically amplified apoptotic signals in GC cells, which may overcome the drug resistance of the BCL-2 inhibitor. These findings indicated that morin was a potent and promising agent for GC treatment. Please cite this article as: Wang Y, Sun XY, Ma FQ, Ren MM, Zhao RH, Qin MM, Zhu XH, Xu Y, Cao ND, Chen YY, Dong TG, Pan YF, Zhao AG. Morin inhibits ubiquitination degradation of BCL-2 associated agonist of cell death and synergizes with BCL-2 inhibitor in gastric cancer cells. J Integr Med. 2025; 23(3): 320-332.
Humans ; Flavonoids/therapeutic use* ; Stomach Neoplasms/pathology* ; Animals ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; Cell Line, Tumor ; Apoptosis/drug effects* ; Cell Proliferation/drug effects* ; Ubiquitination/drug effects* ; Mice ; Drug Synergism ; Mice, Inbred BALB C ; Mice, Nude ; Xenograft Model Antitumor Assays ; Flavones

Traditional Chinese medicine (TCM) is a well-accepted therapy for atopic dermatitis (AD). However, there are currently no evidence-based guidelines integrating TCM and Western medicine for the treatment of AD, limiting the clinical application of such combined approaches. Therefore, the China Association of Chinese Medicine initiated the development of the current guideline, focusing on key issues related to the use of TCM in the treatment of AD. This guideline was developed in accordance with the principles of the guideline formulation manual published by the World Health Organization. A comprehensive review of the literature on the combined use of TCM and Western medicine to treat AD was conducted. The findings were extensively discussed by experts in dermatology and pharmacy with expertise in both TCM and Western medicine. This guideline comprises 23 recommendations across seven major areas, including TCM syndrome differentiation and classification of AD, principles and application scenarios of TCM combined with Western medicine for treating AD, outcome indicators for evaluating clinical efficacy of AD treatment, integration of TCM pattern classification and Western medicine across disease stages, daily management of AD, the use of internal TCM therapies and proprietary Chinese medicines, and TCM external treatments. Please cite this article as: Du XR, Wu MY, Tao MC, Lin Y, Gu CY, Wu MF, Cao Y, Chen DC, Li W, Wang HW, Wang Y, Wang Y, Lu HZ, Liu X, Su XF, Li FL. Clinical practice guidelines for the diagnosis and treatment of atopic dermatitis with integrative traditional Chinese and Western medicine. J Integr Med. 2025; 23(6):641-653.
Dermatitis, Atopic/drug therapy* ; Humans ; Medicine, Chinese Traditional/methods* ; Integrative Medicine ; Drugs, Chinese Herbal/therapeutic use* ; Practice Guidelines as Topic

Objective Based on nuclear factor E2 associated factor 2(NRF2)/PTEN induced hypothesized kinase 1(PINK1)pathway,explored the ameliorating effect of rutaecarpine on chronic obstructive pulmonary disease(COPD)rats and its repairing effect on airway epithelial barrier.Methods COPD rat model were established by smoke combined with airway infusion of lipopolysaccharide;and were randomly divided into model group,control group,and experimental-L group,experimental-M group,experimental-H group,10 rats per group.Another 10 normal rats were selected as the normal group.Experimental-L,-M,-H groups were intraperitoneally injected 15,30 and 60 mg·mL-1 rutaecarpine at the dose of 10 mL·kg-1,respectively.The control group was received 4.05 x 10-2 mg·mL-1 prednisone acetate at the dose of 10 mL·kg-1 by gavage.The normal and model groups were given 0.9％NaCl by intraperitoneal injection.Six groups were treated for 28 days with once a day.The first second forced expiratory volume(FEV1)and forced vital capacity(FVC)of rats were measured by minor animal lung function tester.The levels of interleukin(IL)and interferon-γ(INF-γ)in alveolar lavage fluid were determined by enzyme-linked immunosorbent assay method.The expression levels of PINK1 and NRF2 proteins in the lung tissue were determined by Western blot.Results The levels of FEV1 in the experimental-M group,experimental-H group,control group,model group and normal group were(5.17±0.16),(6.36±0.12),(5.06±0.07),(2.24±0.20)and(6.84±0.11)mL;the levels of FVC were(6.71±0.13),(7.56±0.12),(6.81±0.07),(4.46±0.14)and(7.92±0.11)mL;the levels of IL-12 in alveolar lavage fluid were(7.08±0.51),(9.03±0.54),(7.92±0.79),(3.61±1.01)and(10.15±0.82)pg·mL-1;the levels of IL-9 in alveolar lavage fluid were(22.49±2.27),(15.02±1.41),(17.47±1.84),(38.72±1.28)and(11.78±0.94)pg·mL-1;the levels of INF-γ in alveolar lavage fluid were(13.18±0.54),(16.25±0.60),(15.23±0.43),(6.97±0.89)and(17.22±1.15)pg·mL-1;the relative expression levels of PINK1 protein were 1.10±0.06,1.30±0.09,1.18±0.15,0.42±0.03 and 1.61±0.05;the relative expression levels of NRF2 protein were 0.91±0.05,1.46±0.03,1.35±0.07,0.53±0.07 and 1.64±0.11,respectively.The differences of above indexes were statistically significant between the experimental-M group,experimental-H group,control group and the model group(all P＜0.05).Conclusion Rutaecarpine can inhibit inflammation,improve lung function and repair airway epithelial barrier in COPD rats,and its mechanism may be related to regulating NRF2/PINK1 pathway.

PURPOSE: This study aims to identify the prevalence and risk factors of military training-related abdominal injuries and help plan and conduct training properly. METHODS: This questionnaire survey study was conducted from October 2021 to May 2022 among military personnel from 6 military units and 8 military medical centers and participants' medical records were consulted to identify the training-related abdominal injuries. All the military personnel who ever participated in military training were included. Those who refused to participate in this study or provided an incomplete questionnaire were excluded. The questionnaire collected demographic information, type of abdominal injury, frequency, training subjects, triggers, treatment, and training disturbance. Chi-square test and t-test were used to compare baseline information. Univariate and multivariate regression analyses were used to explore the risk factors associated with military training-related abdominal injuries. RESULTS: A total of 3058 participants were involved in this study, among which 1797 (58.8%) had suffered training-related abdominal injuries (the mean age was 24.3 years and the service time was 5.6 years), while 1261 (41.2%) had no training-related abdominal injuries (the mean age was 23.1 years and the service time was 4.3 years). There were 546 injured patients (30.4%) suspended the training and 84 (4.6%) needed to be referred to higher-level hospitals. The most common triggers included inadequate warm-up, fatigue, and intense training. The training subjects with the most abdominal injuries were long-distance running (589, 32.8%). Civil servants had the highest rate of abdominal trauma (17.1%). Age ≥ 25 years, military service ≥ 3 years, poor sleep status, and previous abdominal history were independent risk factors for training-related abdominal injury. CONCLUSION More than half of the military personnel have suffered military training-related abdominal injuries. Inadequate warm-up, fatigue, and high training intensity are the most common inducing factors. Scientific and proper training should be conducted according to the factors causing abdominal injuries.
Humans ; Military Personnel ; Risk Factors ; Prevalence ; Male ; Abdominal Injuries/etiology* ; Female ; Adult ; Surveys and Questionnaires ; Young Adult

OBJECTIVES: To analyze the association between biological aging markers (phenotypic age and phenotypic age acceleration) and valvular heart diseases. METHODS: Research subjects who met the inclusion and exclusion criteria were selected from the UK Biobank from 2006 to 2010. The phenotypic age and phenotypic age acceleration were calculated. Cox multivariate analysis was used to examine the relationship between the aging markers and valvular heart diseases. Sensitivity analysis was conducted by removing missing values and subgroup analysis. The predictive accuracy of phenotypic age and phenotypic age acceleration for valvular heart diseases was analyzed using receiver operating characteristic (ROC) curves, and a clinical decision curve was generated based on logistic regression. RESULTS: A total of 411 687 subjects were included in the study, among whom there were 14 258 patients with valvular heart diseases. The overall median follow-up time was 12.80 years, the median follow-up time for patients with non-rheumatic aortic valve diseases (n=5238), non-rheumatic mitral valve diseases (n=4558), and non-rheumatic tricuspid valve diseases (n=411) were 12.82 years, 12.83 years and 12.84 years, respectively. After adjusting for demographic factors (gender, race, education, Townsend deprivation index), anthropometric factors (body mass index), lifestyle factors (smoking, alcohol consumption, Dietary Approaches to Stop Hypertension score), hypertension and hyperlipidemia, Cox multivariate analysis showed phenotypic age and phenotypic age acceleration were independent risk factors for valvular heart diseases, including non-rheumatic aortic valve diseases, non-rheumatic mitral valve diseases, and non-rheumatic tricuspid valve diseases (phenotypic age: corrected HR=1.04, P<0.01; phenotypic age acceleration: corrected HR=1.03, P<0.01), which was also confirmed by sensitivity analysis. ROC curves and clinical decision curves demonstrated that compared with the phenotypic age acceleration, phenotypic age had higher accuracy (the areas and the curves were 0.721 and 0.599) and higher net benefit in predicting valvular heart diseases. Moreover, compared with a single indicator, the combination of the two indicators had higher accuracy (the area under the curve was 0.725) and higher net benefit. CONCLUSIONS Phenotypic age and phenotypic age acceleration,as markers of biological aging, are independent risk factors for valvular heart diseases. Compared with phenotypic age acceleration, phenotypic age has a greater advantage in predicting valvular heart diseases. Overall, the combination of the two indicators offers a more effective approach for predicting valvular heart diseases.
Humans ; Male ; Female ; Heart Valve Diseases/epidemiology* ; Middle Aged ; Aged ; Aging ; Adult ; Biomarkers ; Phenotype ; Risk Factors ; Aged, 80 and over

As a new generation of time-of-flight mass spectrometry,multiple-reflection time-of-flight mass spectrometry(MR-TOF-MS)has been increasingly applied in the fields such as nuclear physics,chemistry,and biology due to its ultra-high resolution and rapid analysis capabilities.However,the analytical performance of MR-TOF-MS largely depends on the ion bunch state entering the mass analyzer.In this study,a rectangular ion trap(RIT)was developed,designed and processed using printed circuit board technology,as an ion accumulating and focusing device for MR-TOF mass analyzer.Compared to traditional ion traps composed of two sets of planar electrodes,this RIT had higher voltage utilization efficiency,resulting in more efficient ion collection and focusing.The ions were cooled to a sufficiently small bunch for precise mass measurement with MR-TOF-MS mass spectrometry in only 1 ms of cooling time in the RIT,then orthogonally ejected to the MR-TOF mass spectrometer for mass analysis.Experimental results indicated that the working cycle,ion flux,and ion focusing state of the RIT fully met the requirements of the MR-TOF mass analyzer.When coupled with the MR-TOF mass analyzer,the RIT enabled MR-TOF-MS to achieve a mass resolution of 1.5×105.

An assay was established for detection of toxigenic Clostridioides difficile by combining microfluidic chip analysis with immunochromatography,and its performance was evaluated and compared with those of the Xpert C.difficile/Epi and VIDAS CD AB tests.Primer pairs were designed according to the tcdB and tpi genes in C.difficile.The specificity,limit of detection,reproducibility,and stability were evaluated.A total of 215 stool samples from patients with diarrhea were collected and tested in parallel with the Xpert C.difficile/Epi,VIDAS CDAB,and our assay.C.difficile was isolated from samples,and the tcdB gene was identified when discrepant results were obtained from the three above assays.Our assay showed no cross-reaction with other diarrhea-associated pathogens.Its reproducibility was 100％in testing of two standard plasmids containing tcdB and tpi genes at two concentrations(105 and 102 copies/μL).Two standard plasmids were detected after the PCR and immunochromatography reagents had been stored for 3,6,9,and 12 months,and all the results were posi-tive.The limit of detection was 10 copies/μL for toxigenic C.difficile.Testing of 33 samples positive for C.difficile with our assay(33/215,15.3％)yielded findings statistically coherent with those of the Xpert C.difficile/Epi test(kappa value=0.965).The sensitivity,specificity,positive predictive value,and negative predictive value of our assay,with respect to Xpert C.difficile/Epi as the standard,were 94.3％,100.0％,100.0％,and 98.9％;these values were significantly higher than those of VIDAS CDAB(60.0％,98.9％,91.3％,and 92.7％)(Kappa=0.714,OR=157.50,95％CI:62.03-847.28,P=0.013).In conclusion,our newly developed assay is specific,stable,and reproducible,and may be used for rapid and accu-rate detection of toxigenic C.difficile.The assay could be used for C.difficile infection screening in outpatient and emergen-cy,community medical service center,and epidemiological settings.

Objective Based on nuclear factor E2 associated factor 2(NRF2)/PTEN induced hypothesized kinase 1(PINK1)pathway,explored the ameliorating effect of rutaecarpine on chronic obstructive pulmonary disease(COPD)rats and its repairing effect on airway epithelial barrier.Methods COPD rat model were established by smoke combined with airway infusion of lipopolysaccharide;and were randomly divided into model group,control group,and experimental-L group,experimental-M group,experimental-H group,10 rats per group.Another 10 normal rats were selected as the normal group.Experimental-L,-M,-H groups were intraperitoneally injected 15,30 and 60 mg·mL-1 rutaecarpine at the dose of 10 mL·kg-1,respectively.The control group was received 4.05 x 10-2 mg·mL-1 prednisone acetate at the dose of 10 mL·kg-1 by gavage.The normal and model groups were given 0.9％NaCl by intraperitoneal injection.Six groups were treated for 28 days with once a day.The first second forced expiratory volume(FEV1)and forced vital capacity(FVC)of rats were measured by minor animal lung function tester.The levels of interleukin(IL)and interferon-γ(INF-γ)in alveolar lavage fluid were determined by enzyme-linked immunosorbent assay method.The expression levels of PINK1 and NRF2 proteins in the lung tissue were determined by Western blot.Results The levels of FEV1 in the experimental-M group,experimental-H group,control group,model group and normal group were(5.17±0.16),(6.36±0.12),(5.06±0.07),(2.24±0.20)and(6.84±0.11)mL;the levels of FVC were(6.71±0.13),(7.56±0.12),(6.81±0.07),(4.46±0.14)and(7.92±0.11)mL;the levels of IL-12 in alveolar lavage fluid were(7.08±0.51),(9.03±0.54),(7.92±0.79),(3.61±1.01)and(10.15±0.82)pg·mL-1;the levels of IL-9 in alveolar lavage fluid were(22.49±2.27),(15.02±1.41),(17.47±1.84),(38.72±1.28)and(11.78±0.94)pg·mL-1;the levels of INF-γ in alveolar lavage fluid were(13.18±0.54),(16.25±0.60),(15.23±0.43),(6.97±0.89)and(17.22±1.15)pg·mL-1;the relative expression levels of PINK1 protein were 1.10±0.06,1.30±0.09,1.18±0.15,0.42±0.03 and 1.61±0.05;the relative expression levels of NRF2 protein were 0.91±0.05,1.46±0.03,1.35±0.07,0.53±0.07 and 1.64±0.11,respectively.The differences of above indexes were statistically significant between the experimental-M group,experimental-H group,control group and the model group(all P＜0.05).Conclusion Rutaecarpine can inhibit inflammation,improve lung function and repair airway epithelial barrier in COPD rats,and its mechanism may be related to regulating NRF2/PINK1 pathway.