The neonatal mammalian heart has a remarkable regenerative capacity, while the adult heart has difficulty to regenerate. A metabolic reprogramming from glycolysis to fatty acid oxidation occurs along with the loss of cardiomyocyte proliferative capacity shortly after birth. In this study, we sought to determine if and how metabolic reprogramming regulates cardiomyocyte proliferation. Reversing metabolic reprogramming by carnitine palmitoyltransferase 1 (CPT1) inhibition, using cardiac-specific Cpt1a and Cpt1b knockout mice promoted cardiomyocyte proliferation and improved cardiac function post-myocardial infarction. The inhibition of CPT1 is of pharmacological significance because those protective effects were replicated by etomoxir, a CPT1 inhibitor. CPT1 inhibition, by decreasing poly(ADP-ribose) polymerase 1 expression, reduced ADP-ribosylation of dual-specificity phosphatase 1 in cardiomyocytes, leading to decreased p38 MAPK phosphorylation, and stimulation of cardiomyocyte proliferation. Our present study indicates that reversing metabolic reprogramming is an effective strategy to stimulate adult cardiomyocyte proliferation. CPT1 is a potential therapeutic target for promoting heart regeneration and myocardial infarction treatment.

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 the association between long-term glycemic control and cerebral infarction risk in patients with diabetes through a large-scale cohort study. METHODS: This prospective, community-based cohort study included 12,054 patients with diabetes. From 2006 to 2012, 38,272 fasting blood glucose (FBG) measurements were obtained from these participants. FBG trajectory patterns were generated using latent mixture modelling. Cox proportional hazards models were applied to assess the subsequent risk of cerebral infarction associated with different FBG trajectory patterns. RESULTS: At baseline, the mean age of the participants was 55.2 years. Four distinct FBG trajectories were identified based on FBG concentrations and their changes over the 6-year follow-up period. After a median follow-up of 6.9 years, 786 cerebral infarction events were recorded. Different trajectory patterns were associated with significantly varied outcome risks (Log-Rank P < 0.001). Compared with the low-stability group, Hazard Ratio ( HR) adjusted for potential confounders were 1.37 for the moderate-increasing group, 1.23 for the elevated-decreasing group, and 2.08 for the elevated-stable group. CONCLUSION Sustained high FBG levels were found to play a critical role in the development of ischemic stroke among patients with diabetes. Controlling FBG levels may reduce the risk of cerebral infarction.
Humans ; Cerebral Infarction/blood* ; Middle Aged ; Male ; Female ; Blood Glucose/analysis* ; Fasting/blood* ; Aged ; Prospective Studies ; Risk Factors ; Diabetes Mellitus/blood* ; Adult ; Proportional Hazards Models

AIM To study the chemical constituents from the bark of Toona sinensis(A.Juss.)Roem and their anti-inflammatory activity.METHODS Silica gel,RP-18 reverse phase silica gel and HPLC were used for isolation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.Their anti-inflammatory activity were evaluated by RAW264.7 model.RESULTS Sixteen compounds were isolated and identified as(9Z)-18-hydroxyo-ctadec-9-en-4,6-diyn-3-one(1),toonapolyyne C(2),(9S,10E,16R)-octadec-10-ene-12,14-diyne-1,9,16-triol(3),(9S,10E,16R)-9,16-dihydroxyoctadec-10-ene-12,14-diy-n-1-yl acetate(4),toonapolyyne A(5),toonasindiyne B(6),scopoletin(7),sco-parone(8),3-O-acetyl(-)-epicatechin(9),p-ethoxyacetanilide(10),kulonic acid(11),β-sitosterol-3-O-β-D-glucopyranoside(12),vanillic acid(13),cleomiscosin A(14),(-)-isolariciresinol(15),resveratrol(16).The IC50 values of compounds 3-4,6,11-13 for NO were 6.90,10.49,20.03,9.49,18.34,24.36 μmol/L,respectively.CONCLUSION Compound 1 is a new polyacetylene,8-16 are isolated from T.sinensis for the first time.Compounds 3-4,6,11-13 have good anti-inflammatory activity.

Objective:To preliminarily establish a sample clot warning model for coagulation screening tests using 5 machine learning methods.Methods:This cross-sectional study collected 7 401 routine screening test samples from Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, from January 1st, 2015, to August 18th, 2024, including 4 786 clotted (positive) and 2 615 qualified (negative) samples for model development. The dataset was divided into Dataset 1 and Dataset 2 based on a reagent change for APTT in December 2018, with separate models developed for each. An additional 2 493 samples (October 31st to November 8th, 2024) were used to evaluate consistency between the model and manual assessment, while 23 200 samples (October 17th to December 31st, 2024) were used for assessing real-world predictive performance. Five machine learning algorithms were employed to develop the clot prediction model: logistic regression (LR), random forest (RF), extreme gradient boosting (XGBoost), naive bayes (NB), and artificial neural network (ANN), with the ANN model constructed using two different hidden layer and neuron parameter settings. Model selection was based on AUC, accuracy, sensitivity, specificity, F1-score, PPV, and NPV, with the optimal model integrated into the LIS for validation.Results:Among the six models using 5 machine learning algorithms, XGBoost demonstrated the highest performance (AUC=0.961, sensitivity=0.945, F1-score=0.934) and robustness to reagent changes ( Z=-1.333, P=0.113). When deployed, the differences between the model's predictions and manual pre-judgment were statistically significant ( Z=-5.289 to 8.933, all P<0.01). The predictive efficacy indices AUC (95% CI), sensitivity, specificity, and accuracy of the XGBoost model deployed in real-world operation of the LIS were 0.939 (0.918—0.960), 0.958, 0.921, and 0.921 respectively. Conclusion:In this study, a clot warning model for coagulation screening samples was established based on the XGBoost algorithm, and its prediction efficacy is good, providing a foundation for intelligent pre-analytical quality control for coagulation screening tests.

Objective : To develop and to validate a combined model integrating18F-FDG PET/CT radiomics with clinical features to distinguish between lymphoma and lymphoid inflammatory hyperplasia. Methods : A retrospective study was conducted on a cohort of 232 patients diagnosed with lymphoma or lymphoid inflammatory hyperplasia. Comparative analyses of clinical and traditional imaging indicators were performed to identify inter-group differences. The clinical features were delineated and extracted using medical software including 3D-Slicer and Lifex. Selection of the features was performed to construct a PET/CT-based radiomics Logistic model, with a combined model integrating PET/CT with clinical features then used to evaluate the discriminative efficacy of these models. Results: Analysis of inter-group differences indicated that age, CTmean, and metabolic tumor volume(MTV)were effective for differentiating between lymphoma and lymphoid inflammatory hyperplasia(P<0.05). The PET/CT-based radiomics Logistic model differentiated between lymphoma and lymphoid inflammatory hyperplasia, with an area under curve(AUC) of 0.924(95%CI: 0.884-0.960) and 0.863(95%CI: 0.774-0.939) in the training and testing cohorts, respectively. The integrated Logistic model that combined PET/CT-based radiomics with clinical features to distinguish between lymphoma and lymphoid inflammatory hyperplasia achieved an AUC of 0.933(95%CI: 0.889-0.969) in the training cohort and 0.884(95%CI: 0.792-0.964) in the testing cohort. Decision curve analysis(DCA) demonstrated that the integrated model provided the greatest clinical net benefit. Conclusion The hybrid model integrating18F-FDG PET/CT radiomics with clinical features shows robust diagnostic efficacy to distinguish between lymphoma and lymphoid inflammatory hyperplasia.

Objective:To explore the diagnostic value of total bilirubin to albumin ratio(B/A ratio)combined with alpha-fetoprotein(AFP)and abnormal prothrombin induced by vitamin K absence-Ⅱ(PIVKA-Ⅱ)for hepatocellular carcinoma(HCC).Methods:35 HCC patients(HCC group),35 cirrhosis patients(cirrhosis group),35 HCC patients post-transcatheter arterial chemoembolization(TACE)(TACE postoperative group),and 35 healthy volunteers(healthy control group)were selected in our hospital from October 2023 to October 2024.The serum B/A ratio,AFP,and PIVKA-Ⅱ levles were measured and compared across the groups.The correlations between serum B/A ratio,AFP,and PIVKA-Ⅱ in the HCC group were analyzed.The B/A ratio,AFP,PIVKA-Ⅱ were compared across different clinical and pathological features in HCC patients.The serum B/A ratio,AFP,and PIVKA-Ⅱ levels were compared pre and post operation in the HCC group.The diagnostic value of B/A ratio,AFP,PIVKA-Ⅱ alone and in combination for HCC were analyzed by receiver operating characteristic(ROC)curves.Results:The serum B/A ratio,AFP,and PIVKA-Ⅱ levels in HCC group and TACE postoperative group were significantly higher than those in the cirrhosis group and healthy control group,and the HCC group was higher than that in the TACE postoperative group(P＜0.05).Pearson correlation analysis results showed that,B/A ratio in the HCC group was positively correlated with AFP(r=0.352,P=0.001),B/A ratio was positively correlated with PIVKA-Ⅱ(r=0.327,P=0.003),and AFP was positively correlated with PIVKA-Ⅱ(r=0.285,P=0.008).Higher TNM stage,larger tumor diameter,presence of vascular invasion,and lower differentiation degree of HCC patients,who had higher B/A ratio,AFP,and PIVKA-Ⅱ levels(P＜0.05).Serum B/A ratio,AFP,and PIVKA-Ⅱ levels in the HCC group post operation were significantly lower than those in pre operation(P＜0.05).ROC curve analysis results showed that,when B/A ratio,AFP,and PIVKA-Ⅱ were detected separately,the area under the curve(AUC)was 0.785,0.756,and 0.802,respectively.The AUC for joint detection was 0.925.The AUC in combination was greater than that of individual detection of each indicator.Conclusion:The combination of B/A ratio,AFP,and PIVKA-Ⅱ testing significantly improves the diagnostic efficiency for HCC,which is worthy of clinical application.

Objective:To analyze the factors associated with prolonged hospital stay in patients with mycoplasma pneumoniae pneumonia (MPP) and construct an early identification model.Methods:A case-control study was conducted on 503 newly diagnosed pediatric patients with MPP who received treatment at Suzhou Municipal Hospital from April to December 2023. Clinical data were collected. Patients were divided into an observation group ( n = 240, hospital stay > 8 days) and a control group ( n = 263, hospital stay ≤ 8 days). Logistic regression analysis was used to identify the independent factors that affect hospital stay in the observation group. A Nomogram model was constructed. Results:The incidence of lobar pneumonia in the observation group was significantly higher than that in the control group [62.5% (150/240) vs. 22.4% (59/263), χ2 = 82.94, P < 0.001]. The mean age of the observation group was significantly higher than that in the control group [5.0 (3.0, 7.0) years vs. 4.0 (2.0, 6.0) years, Z = 2.40, P = 0.016]. The hypersensitive C-reactive protein level in the observation group was significantly higher than that in the control group [10.5 (4.8, 22.0) mg/L vs. 6.1 (1.8, 14.2) mg/L, Z = 5.16, P < 0.001]. The ferritin level in the observation group was significantly higher than that in the control group [225.3 (180.9, 271.3) μg/L vs. 177.7 (138.0, 222.0) μg/L, Z = 6.31, P < 0.001]. The albumin level in the observation group was significantly lower than that in the control group [43.7 (41.0, 46.2) g/L vs. 44.4 (42.3, 46.5) g/L, Z = 2.45, P = 0.014]. The total protein level in the observation group was significantly lower than that in the control group [70.0 (66.8, 73.0) g/L vs. 71.5 (67.8, 74.6) g/L, Z = 2.45, P = 0.014]. The lactate dehydrogenase level in the observation group was significantly higher than that in the control group [337.5 (301.5, 391.8) U/L vs. 291.0 (258.0, 332.3) U/L, Z = 3.28, P = 0.001]. The creatine kinase level in the observation group was significantly lower than that in the control group [77.5 (55.3, 115.8) U/L vs. 89.0 (65.0, 126.0) U/L, Z = 2.75, P = 0.006]. The fibrinogen level in the observation group was significantly higher than that in the control group [4.3 (3.4, 4.8) g/L vs. 3.8 (3.0, 4.6) g/L, Z = 4.17, P < 0.001]. After performing univariate binary logistic regression using the glm method to screen variables, multivariate binary logistic regression was conducted. The results showed that the presence of lobar pneumonia, higher levels of hypersensitive C-reactive protein, ferritin, and lactate dehydrogenase were independent risk factors for prolonged hospital stays in children with MPP [ OR (95% CI): 3.803 (2.029,7.129), 0.986 (0.974,0.998), 0.994 (0.990,0.998), 0.989 (0.985,0.993), P < 0.001, 0.027, 0.002, < 0.001]. Conclusions:Based on the fundamental clinical laboratory indicators, an early prediction model was constructed for predicting prolonged hospital stay in children with MPP. This model provides a scientific basis for the early assessment of MPP in children and is suitable for broader application.

AIM To study the chemical constituents from the bark of Toona sinensis(A.Juss.)Roem and their anti-inflammatory activity.METHODS Silica gel,RP-18 reverse phase silica gel and HPLC were used for isolation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.Their anti-inflammatory activity were evaluated by RAW264.7 model.RESULTS Sixteen compounds were isolated and identified as(9Z)-18-hydroxyo-ctadec-9-en-4,6-diyn-3-one(1),toonapolyyne C(2),(9S,10E,16R)-octadec-10-ene-12,14-diyne-1,9,16-triol(3),(9S,10E,16R)-9,16-dihydroxyoctadec-10-ene-12,14-diy-n-1-yl acetate(4),toonapolyyne A(5),toonasindiyne B(6),scopoletin(7),sco-parone(8),3-O-acetyl(-)-epicatechin(9),p-ethoxyacetanilide(10),kulonic acid(11),β-sitosterol-3-O-β-D-glucopyranoside(12),vanillic acid(13),cleomiscosin A(14),(-)-isolariciresinol(15),resveratrol(16).The IC50 values of compounds 3-4,6,11-13 for NO were 6.90,10.49,20.03,9.49,18.34,24.36 μmol/L,respectively.CONCLUSION Compound 1 is a new polyacetylene,8-16 are isolated from T.sinensis for the first time.Compounds 3-4,6,11-13 have good anti-inflammatory activity.