This study aimed to characterize and identify the non-volatile components in aqueous and ethanolic extracts of the stems and leaves of Rhododendron tomentosum by using sensitive and efficient ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry(UHPLC-Q-TOF-MS) combined with a self-built information database. By comparing with reference compounds, analyzing fragment ion information, searching relevant literature, and using a self-built information database, 118 compounds were identified from the aqueous and ethanolic extracts of R. tomentosum, including 35 flavonoid glycosides, 15 phenolic glycosides, 12 flavonoids, 7 phenolic acids, 7 phenylethanol glycosides, 6 tannins, 6 phospholipids, 5 coumarins, 5 monoterpene glycosides, 6 triterpenes, 3 fatty acids, and 11 other types of compounds. Among them, 102 compounds were reported in R. tomentosum for the first time, and 36 compounds were identified by comparing them with reference compounds. The chemical components in the ethanolic and aqueous extracts of R. tomentosum leaves and stems showed slight differences, with 84 common chemical components accounting for 71.2% of the total 118 compounds. This study systematically characterized and identified the non-volatile chemical components in the ethanolic and aqueous extracts of R. tomentosum for the first time. The findings provide a reference for active ingredient research, quality control, and product development of R. tomentosum.
Rhododendron/chemistry* ; Chromatography, High Pressure Liquid/methods* ; Drugs, Chinese Herbal/chemistry* ; Mass Spectrometry/methods* ; Plant Leaves/chemistry*

OBJECTIVE: EPF3 is a fibrinolysin monomer isolated and purified from Pheretima vulgaris Chen, an earthworm used in traditional Chinese medicine as Dilong for treating blood stasis syndrome. Its composition, anticoagulant and fibrinolytic activities, and relevant mechanisms have been confirmed through in vitro experiments. However, whether it has antithrombotic effects in vivo and can be absorbed by the gastrointestinal tract is unknown. This study evaluates the antithrombotic effect in zebrafish and investigates the gastrointestinal stability and intestinal absorption mechanism of this protein in vitro. METHODS: The antithrombotic effect of EPF3 in vivo was verified using the zebrafish thrombus model induced by arachidonic acid and FeCl₃. Then, the protein bands of EPF3 incubated with simulated gastric fluid (SGF), simulated intestinal fluid (SIF), and homogenate of Caco-2 cells (HC2C) were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to evaluate its gastrointestinal stability. Finally, the transport behavior and absorption mechanism of EPF3 were studied using Caco-2 cell monolayer. RESULTS: EPF3 could significantly enhance the returned blood volume and blood flow velocity in zebrafish with platelet aggregation thrombus induced by arachidonic acid. It could also prolong the formation time of tail artery thrombus and increase the blood flow velocity in zebrafish with vessel injury thrombus induced by FeCl₃. EPF3 was stable in SIF and HC2C and unstable in SGF. The permeability of EPF3 in Caco-2 monolayer was time-dependent and concentration-dependent. The efflux ratio was less than 1.2 during transport, and the transport behavior was not affected by inhibitors. EPF3 could reversibly reduce the expression of tight junction-related proteins, including zonula occludens-1, occludin, and claudin-1 in Caco-2 cells. CONCLUSION EPF3 could play a thrombolytic and antithrombotic role in zebrafish. It could be transported and absorbed into the intestine through cellular bypass pathway by opening the intestinal epithelium tight junction. This study provides a scientific explanation for the antithrombotic effect of earthworm and provides a basis for the feasibility of subsequent development of EPF3 as an antithrombotic enteric-soluble preparation. Please cite this article as: Zhong WL, Yang JQ, Liu H, Wu YL, Shen HJ, Li PY, Du SY. Antithrombotic effect in zebrafish of a fibrinolytic protein EPF3 from Dilong (Pheretima vulgaris Chen) and its transport mechanism in Caco-2 monolayer through cell bypass pathway. J Integr Med. 2025; 23(4): 415-428.
Animals ; Zebrafish ; Humans ; Caco-2 Cells ; Fibrinolytic Agents/pharmacology* ; Thrombosis/drug therapy* ; Intestinal Absorption

Objective To investigate gender differences and age-related changes in body composition(BC)among Miao adults in Rongshui,Guangxi Province,and to provide the basis for assessing nutritional status and health.Methods With informed consent,630 Miao adults(218 males,412 females)were randomly selected for this study.Body composition was assessed using bioelectrical impedance analysis(BIA).Results Weight,fat-free mass,muscle mass,trunk muscle mass,limb muscle mass,waist-to-hip ratio(WHR),body water,presumtion of bone mass and protein were significantly higher in males than in females.And the fat mass,trunk fat mass,limb fat mass,visceral fat content,subcutaneous fat content and percentage of body fat were significantly higher in females than in males.According to the evaluation of body mass index(BMI)and WHR,the proportion of overweight and obesity of Miao adults was higher than the average level of Miao residents,and their obesity was characterized by central obesity.With age,weight,fat mass,muscle mass,fat-free mass,limb muscle mass,limb fat mass,subcutaneous fat content,percentage of body fat,body water,presumtion of bone mass,and protein of Rongshui Miao adults showed a gradual decreasing trend,while visceral fat content and WHR increased progressively.BMI in male Miao adults,along with BMI,fat mass,trunk fat mass,subcutaneous fat content,percentage of body fat,and body water in female Miao adults,showed a trend of increasing followed by decreasing,peaking at the age of 40-49 years.Conclusion The body composition of Miao adults in Rongshui,Guangxi,exhibits significant gender differences and age-related variation change patterns,which may increase the risk of sarcopenia and metabolic diseases with aging.

Alzheimer's disease (AD), a progressive dementia, is one of the most common neurodegenerative diseases. Clinical trial results of amyloid-β (Aβ) and tau regulators based on the pretext of straightforward amyloid and tau immunotherapy were disappointing. There are currently no effective strategies for slowing the progression of AD. Herein, we spotlight the dysregulation of lipid metabolism, particularly the elevation of ceramides (Cers), as a critical yet underexplored facet of AD pathogenesis. Our study delineates the role of Cers in promoting microglial pyroptosis, a form of programmed cell death distinct from apoptosis and necroptosis, characterized by cellular swelling, and membrane rupture mediated by the NLRP3 inflammasome pathway. Utilizing both in vivo experiments with amyloid precursor protein (APP)/presenilin 1 (PS1) transgenic mice and in vitro assays with BV-2 microglial cells, we investigate the activation of microglial pyroptosis by Cers and its inhibition by icariin (ICA), a flavonoid with known antioxidant and anti-inflammatory properties. Our findings reveal a significant increase in Cers levels and pyroptosis markers (NOD-like receptor family, pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein containing a caspase recruitment domain, caspase-1, gasdermin D (gasdermin D (GSDMD)), and interleukin-18 (IL-18)) in the brains of AD model mice, indicating a direct involvement of Cers in AD pathology through the induction of microglial pyroptosis. Conversely, ICA treatment effectively reduces these pyroptotic markers and Cer levels, thereby attenuating microglial pyroptosis and suggesting a novel therapeutic mechanism of action against AD. This study not only advances our understanding of the pathogenic role of Cers in AD but also introduces ICA as a promising candidate for AD therapy, capable of mitigating neuroinflammation and pyroptosis through the cyclooxygenase-2 (COX-2)-NLRP3 inflammasome-gasdermin D (GSDMD) axis. Our results pave the way for further exploration of Cer metabolism disorders in neurodegenerative diseases and highlight the therapeutic potential of targeting microglial pyroptosis in AD.

Objective To explore the efficacy differences between 3 Tesla magnetic resonance imaging with zero echo time(3T MRI ZTE)and high resolution computed tomography(HRCT)in the detection of pulmonary nodules and the classification diagnosis of the lung imaging reporting and data system(Lung-RADS)in sub-health populations.Methods Clinical and imaging data of 93 patients with pulmonary nodules(126 nodules in total)admitted to some hospital from July to December 2023 were retrospectively analyzed.The 126 nodules were categorized into a benign nodule group(n=51)and a malignant nodule group(n=75)using pathological findings as the gold standard.All the patients underwent examinations by 3T MRI ZTE and HRCT to compare the detection rates of the two measures for pulmonary nodules;the missed and misdiagnosis rates of 3T MRI ZTE,HRCT and Lung-RADS grading were contrasted with the postoperative pathological diagnosis results as the gold standard;comparison analyses of 3T MRI ZTE signs and HRCT signs were performed between the two groups and the patients with different Lung-RADS grades;3T MRI ZTE,HRCT and Lung-RADS grading were compared with the receiver operating characteristic(ROC)curve in terms of diagnosis efficacy for pulmonary nodules,and the consistency analysis was carried out.Results No discernible statistical variation was observed in the detection rates of pulmonary nodules between 3T MRI ZTE and HRCT(P>0.05).Lung-RADS grading had the highest rates of missed diagnosis and misdiagnosis,and 3T MRI ZTE and HRCT had similar detection rates.The malignant nodule group was different from the benign nodule group in the 3T MRI ZTE and HRCT signs in terms of lesion size,spiculation sign,lobulation sign,calcifica-tion,pleural indentation sign,cavity sign,boundary and bronchial cut-off sign,with the differences being statistically signi-ficant(all P<0.05).For the patients of Lung-RADS grade 3,the 3T MRI ZTE and HRCT signs had significant differences in terms of lesion size,spiculation sign,lobulation sign,calcification,pleural indentation sign,cavity sign and bronchial cut-off sign(all P<0.05).For the patients of Lung-RADS grade 4A,the 3T MRI ZTE and HRCT signs had significant differen-ces in terms of lesion size,calcification,boundary and bronchial cut-off sign(all P<0.05).For the patients of Lung-RADS grade 4B,the 3T MRI ZTE and HRCT signs had significant differences in terms of lesion size and calcification(all P<0.05).For the patients of Lung-RADS grade 4X,there were no significant differences found between the 3T MRI ZTE and HRCT signs(all P>0.05).HRCT had the highest sensitivity,specificity,accuracy,AUC value,predictive values and Kappa value for benign and malignant nodules,3T MRI ZTE had the values slightly lower than those of HRCT,and Lung-RADS grading had the lowest values when compared with HRCT and 3T MRI ZTE.Conclusion HRCT and 3T MRI ZTE are complementary for the evaluation of pulmonary nodules,and the differences in imaging signs between them show graded dependence.3T MRI ZTE and HRCT have no significant differences in the detection rate of pulmonary nodules,while HRCT gains advanta-ges in differentiating benign and malignant pulmonary nodules,and references are provided for the screening and clinical early diagnosis of pulmonary nodules.[Chinese Medical Equipment Journal,2025,46(9):52-59]

BACKGROUND:In our previous experiments,it was found that transplantation of young adipose stem cells into aged mice would have weight loss effect and improve the inflammatory state in aged mice. Therefore,we speculate that insulin-like growth factor 1 may play an important role in aging and obesity. OBJECTIVE:To explore the anti-obesity effect of insulin-like growth factor 1 in naturally aging mice. METHODS:(1) Bioinformatics analysis:Sequencing of adipose tissue from obese patients in the GEO database and transcriptomic sequencing of young mouse adipose stem cells and old adipose stem cells were conducted to analyze insulin-like growth factor 1 expression. (2) Animal experiment verification:Six young C57BL/6J mice and twelve aged C57BL/6J mice (20 months old) were selected. Abdominal adipose tissue and serum insulin-like growth factor 1 expression in young and aged mice were examined by ELISA and qRT-PCR. All the 12 aged mice were randomly divided into two groups with 6 mice in each group:the experimental group was given insulin-like growth factor 1 (50 μg/kg) for 4 continuous weeks,while the control group was given the same amount of phosphate buffer saline. The body mass changes of mice were monitored regularly,glucose tolerance was measured at the end of the experiment,and serum inflammatory factors and inflammatory factors in abdominal white adipose tissue of mice were detected by ELISA. Hematoxylin-eosin staining was used to observe pathological changes in the mouse liver,kidney and abdominal white adipose tissue. The mRNA expression levels of inflammatory factors and PI3K-Akt signaling pathway in abdominal white adipose tissue of mice were detected by qRT-PCR.RESULTS AND CONCLUSION:Obese patients showed lowly expressed insulin-like growth factor 1 in adipose tissue,but the expression of insulin-like growth factor 1 increased after weight loss surgery. Insulin-like growth factor 1 expressed lowly in aged adipose stem cells. Insulin-like growth factor 1 also showed a low expression in adipose tissue and serum of aged mice. Injection of insulin-like growth factor 1 protein could significantly reduce the body mass of aged mice and improve insulin resistance. Pathological sections of the liver of aged mice revealed fat accumulation. After injection of insulin-like growth factor 1 protein,fat accumulation was significantly improved and the size of fat droplets in adipose tissue was significantly reduced. Insulin-like growth factor 1 injection significantly reduced the expression levels of serum tumor necrosis factor α,interleukin 1β,and interleukin 6 in aged mice,and significantly increased the expression of PI3K-AKT signaling pathway in adipose tissue of aged mice. To conclude,exogenous insulin-like growth factor 1 can reduce body mass,reduce fat droplet size in adipose tissue,and improve liver fat accumulation in aged mice,thereby improving their inflammatory status. Exogenous insulin-like growth factor 1 may activate the PI3K-AKT signaling pathway to improve the inflammatory symptoms in aged mice,thereby improving obesity in naturally aging mice.

Alzheimer's disease(AD),a progressive dementia,is one of the most common neurodegenerative dis-eases.Clinical trial results of amyloid-β(Aβ)and tau regulators based on the pretext of straightforward amyloid and tau immunotherapy were disappointing.There are currently no effective strategies for slowing the progression of AD.Herein,we spotlight the dysregulation of lipid metabolism,particularly the elevation of ceramides(Cers),as a critical yet underexplored facet of AD pathogenesis.Our study delineates the role of Cers in promoting microglial pyroptosis,a form of programmed cell death distinct from apoptosis and necroptosis,characterized by cellular swelling,and membrane rupture mediated by the NLRP3 inflammasome pathway.Utilizing both in vivo experiments with amyloid precursor protein(APP)/presenilin 1(PS1)transgenic mice and in vitro assays with BV-2 microglial cells,we investigate the activation of microglial pyroptosis by Cers and its inhibition by icariin(ICA),a flavonoid with known antioxidant and anti-inflammatory properties.Our findings reveal a significant increase in Cers levels and pyroptosis markers(NOD-like receptor family,pyrin domain containing 3(NLRP3),apoptosis-associated speck-like protein containing a caspase recruitment domain,caspase-1,gasdermin D(GSDMD),and interleukin-18(IL-18))in the brains of AD model mice,indicating a direct involvement of Cers in AD pathology through the induction of microglial pyroptosis.Conversely,ICA treatment effec-tively reduces these pyroptotic markers and Cer levels,thereby attenuating microglial pyroptosis and suggesting a novel therapeutic mechanism of action against AD.This study not only advances our un-derstanding of the pathogenic role of Cers in AD but also introduces ICA as a promising candidate for AD therapy,capable of mitigating neuroinflammation and pyroptosis through the cyclooxygenase-2(COX-2)-NLRP3 inflammasome-gasdermin D(GSDMD)axis.Our results pave the way for further exploration of Cer metabolism disorders in neurodegenerative diseases and highlight the therapeutic potential of tar-geting microglial pyroptosis in AD.

Objective:To investigate the risk factors influencing mortality in neonates undergoing continuous renal replacement therapy (CRRT).Methods:This retrospective study included 34 neonates with a corrected age of≤28 days who received CRRT at the Affiliated Children's Hospital of Xiangya School of Medicine, Central South University, from January 2019 to December 2023. The neonates were divided into a mortality group ( n=16) and a survival group ( n=18) based on whether they died during CRRT. Pre-CRRT blood biochemical indices, general condition, CRRT treatment modes, parameters, and related complications were analyzed using t-tests, Wilcoxon signed-rank tests, and Chi-square tests. Logistic stepwise regression analysis was used to screen for risk factors associated with CRRT mortality. Results:The mortality rate among the 34 neonates was 48.6% (16/34), with a median CRRT age of 17 days (range: 2-33 days). Eleven neonates (32.3%) were preterm, with the youngest gestational age being 27 weeks and the lowest weight before CRRT initiation being 1 700 g. The mortality group had lower urine output 6-12 hours before CRRT initiation and lower critical illness scores compared to the survival group [0.05 (0.02-1.00) ml/(kg·h) vs. 0.50 (0.20-1.05) ml/(kg·h), (64.50±7.10) scores vs. (77.67±3.65) scores, Z or t values were 10.97 and 3.91, respectively]. However, the vasoactive inotropic score (VIS), proportion of coma, and levels of blood potassium, alanine aminotransferase, aspartate aminotransferase, blood ammonia, blood lactic acid, and activated partial thromboplastin time (APTT) were higher in the mortality group compared to the survival group [ (86.88±15.80) scores vs. (55.56±24.31) scores, 11/16 vs. 1/18, (7.02±1.73) mmol/L vs. (5.88±1.53) mmol/L, 274.55(132.50-664.98) U/L vs. 31.10(19.03-110.70) U/L, 688.20 (449.73-3 618.13) U/L vs. 96.65 (44.15-439.00) U/L, 232.75 (70.33-1 310.85) μmol/L vs.77.70 (49.78-919.05) μmol/L, (11.17±3.36) U/L vs. (7.99±2.67) U/L, and (99.57±39.74) s vs. (60.97±31.25) s, with t, χ2, or Z values of－4.39, 14.81,－2.03,－2.72,－11.81,－3.89,－3.06, and－3.17, respectively] (all P<0.05). Logistic regression analysis revealed that pre-treatment VIS value ( OR=1.150, 95% CI: 1.035-1.278), and blood ammonia level ( OR=1.004, 95% CI: 1.002-1.009) were independent risk factors for mortality (both P<0.05). Conclusions:Neonatal CRRT mortality is associated with pre-treatment VIS scores and blood ammonia levels. Attention should be paid to a rapid decreases in urine output, the intensity of vasopressor support, and elevated levels of blood ammonia, blood lactic acid, transaminases, and APTT at the initiation of treatment.

Objective:To investigate the risk factors influencing mortality in neonates undergoing continuous renal replacement therapy (CRRT).Methods:This retrospective study included 34 neonates with a corrected age of≤28 days who received CRRT at the Affiliated Children's Hospital of Xiangya School of Medicine, Central South University, from January 2019 to December 2023. The neonates were divided into a mortality group ( n=16) and a survival group ( n=18) based on whether they died during CRRT. Pre-CRRT blood biochemical indices, general condition, CRRT treatment modes, parameters, and related complications were analyzed using t-tests, Wilcoxon signed-rank tests, and Chi-square tests. Logistic stepwise regression analysis was used to screen for risk factors associated with CRRT mortality. Results:The mortality rate among the 34 neonates was 48.6% (16/34), with a median CRRT age of 17 days (range: 2-33 days). Eleven neonates (32.3%) were preterm, with the youngest gestational age being 27 weeks and the lowest weight before CRRT initiation being 1 700 g. The mortality group had lower urine output 6-12 hours before CRRT initiation and lower critical illness scores compared to the survival group [0.05 (0.02-1.00) ml/(kg·h) vs. 0.50 (0.20-1.05) ml/(kg·h), (64.50±7.10) scores vs. (77.67±3.65) scores, Z or t values were 10.97 and 3.91, respectively]. However, the vasoactive inotropic score (VIS), proportion of coma, and levels of blood potassium, alanine aminotransferase, aspartate aminotransferase, blood ammonia, blood lactic acid, and activated partial thromboplastin time (APTT) were higher in the mortality group compared to the survival group [ (86.88±15.80) scores vs. (55.56±24.31) scores, 11/16 vs. 1/18, (7.02±1.73) mmol/L vs. (5.88±1.53) mmol/L, 274.55(132.50-664.98) U/L vs. 31.10(19.03-110.70) U/L, 688.20 (449.73-3 618.13) U/L vs. 96.65 (44.15-439.00) U/L, 232.75 (70.33-1 310.85) μmol/L vs.77.70 (49.78-919.05) μmol/L, (11.17±3.36) U/L vs. (7.99±2.67) U/L, and (99.57±39.74) s vs. (60.97±31.25) s, with t, χ2, or Z values of－4.39, 14.81,－2.03,－2.72,－11.81,－3.89,－3.06, and－3.17, respectively] (all P<0.05). Logistic regression analysis revealed that pre-treatment VIS value ( OR=1.150, 95% CI: 1.035-1.278), and blood ammonia level ( OR=1.004, 95% CI: 1.002-1.009) were independent risk factors for mortality (both P<0.05). Conclusions:Neonatal CRRT mortality is associated with pre-treatment VIS scores and blood ammonia levels. Attention should be paid to a rapid decreases in urine output, the intensity of vasopressor support, and elevated levels of blood ammonia, blood lactic acid, transaminases, and APTT at the initiation of treatment.

Objective To explore the efficacy differences between 3 Tesla magnetic resonance imaging with zero echo time(3T MRI ZTE)and high resolution computed tomography(HRCT)in the detection of pulmonary nodules and the classification diagnosis of the lung imaging reporting and data system(Lung-RADS)in sub-health populations.Methods Clinical and imaging data of 93 patients with pulmonary nodules(126 nodules in total)admitted to some hospital from July to December 2023 were retrospectively analyzed.The 126 nodules were categorized into a benign nodule group(n=51)and a malignant nodule group(n=75)using pathological findings as the gold standard.All the patients underwent examinations by 3T MRI ZTE and HRCT to compare the detection rates of the two measures for pulmonary nodules;the missed and misdiagnosis rates of 3T MRI ZTE,HRCT and Lung-RADS grading were contrasted with the postoperative pathological diagnosis results as the gold standard;comparison analyses of 3T MRI ZTE signs and HRCT signs were performed between the two groups and the patients with different Lung-RADS grades;3T MRI ZTE,HRCT and Lung-RADS grading were compared with the receiver operating characteristic(ROC)curve in terms of diagnosis efficacy for pulmonary nodules,and the consistency analysis was carried out.Results No discernible statistical variation was observed in the detection rates of pulmonary nodules between 3T MRI ZTE and HRCT(P>0.05).Lung-RADS grading had the highest rates of missed diagnosis and misdiagnosis,and 3T MRI ZTE and HRCT had similar detection rates.The malignant nodule group was different from the benign nodule group in the 3T MRI ZTE and HRCT signs in terms of lesion size,spiculation sign,lobulation sign,calcifica-tion,pleural indentation sign,cavity sign,boundary and bronchial cut-off sign,with the differences being statistically signi-ficant(all P<0.05).For the patients of Lung-RADS grade 3,the 3T MRI ZTE and HRCT signs had significant differences in terms of lesion size,spiculation sign,lobulation sign,calcification,pleural indentation sign,cavity sign and bronchial cut-off sign(all P<0.05).For the patients of Lung-RADS grade 4A,the 3T MRI ZTE and HRCT signs had significant differen-ces in terms of lesion size,calcification,boundary and bronchial cut-off sign(all P<0.05).For the patients of Lung-RADS grade 4B,the 3T MRI ZTE and HRCT signs had significant differences in terms of lesion size and calcification(all P<0.05).For the patients of Lung-RADS grade 4X,there were no significant differences found between the 3T MRI ZTE and HRCT signs(all P>0.05).HRCT had the highest sensitivity,specificity,accuracy,AUC value,predictive values and Kappa value for benign and malignant nodules,3T MRI ZTE had the values slightly lower than those of HRCT,and Lung-RADS grading had the lowest values when compared with HRCT and 3T MRI ZTE.Conclusion HRCT and 3T MRI ZTE are complementary for the evaluation of pulmonary nodules,and the differences in imaging signs between them show graded dependence.3T MRI ZTE and HRCT have no significant differences in the detection rate of pulmonary nodules,while HRCT gains advanta-ges in differentiating benign and malignant pulmonary nodules,and references are provided for the screening and clinical early diagnosis of pulmonary nodules.[Chinese Medical Equipment Journal,2025,46(9):52-59]