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 explore the effect of home-based online preoperative rehabilitation training on early postoperative recovery in patients with pulmonary nodules.Methods:This study is a randomized controlled study. Fifty patients with pulmonary nodules scheduled for elective surgery in Zhongshan Hospital from January 2023 to September 2023 were randomly divided into the control group and the study group ( n=25 in each group). Patients in the study group carried out home pre-rehabilitation training guided by mobile App before operation, including breathing training, resistance exercise, nutritional intervention and psychological counseling; there was no intervention for patients in the control group while waiting for operation. The lung function, 6-min walking distance, psychological status, nutritional status, postoperative complications, catheterization time, hospitalization time, hospitalization expenses and complications were compared between the two groups before and after operation. Results:There were no significant differences in age, sex, BMI, heart rate, respiratory rate, blood pressure, pulmonary function, nutritional status, location of pulmonary nodules, anesthesia mode, operation modality and surgical pathology results between two groups(all P>0.05). Compared with the control group, the study group had significantly improved forced vital capacity at admission, 1d and 30d after surgery ( Z=-2.338, Z=-2.203, Z=-2.348, all P<0.05), forced expiratory volume in one second (FEV1) ( Z=-3.241, Z=-3.262, Z=-3.564, all P<0.05), and the ratio of forced vital capacity (FVC) to FEV1 ( t=-3.186, t=-2.904, and t=-4.304, all P<0.05), increased 6 min walking distance ( t=-4.466, t=-4.144, t=-4.907, all P<0.05), improved anxiety ( Z=-3.164, Z=-4.425, Z=-3.622, all P<0.05), decreased depression ( Z=-2.598, Z=-3.799, Z=-3.439, all P<0.05); and improved nutritional status at admission and 1d after surgery ( Z=-3.061, Z=-3.648, all P<0.05). The incidence of postoperative complications was decreased ( χ 2=4.878, P<0.05), the time of catheterization was decreased ( Z=-4.056, P<0.001), the length of hospital stay was shortened ( Z=-4.345, P<0.001), and the cost of hospitalization was reduced ( Z=-2.590, P<0.05) in the study group. Conclusion:The home-based online prehabilitation training can effectively improve the cardiopulmonary function and physical fitness of patients, reduce the stress of surgery, and contribute to rapid recovery after surgery; it also can shorten the length of postoperative hospital stay and reduce medical costs.

Objective:To explore the effect of home-based online preoperative rehabilitation training on early postoperative recovery in patients with pulmonary nodules.Methods:This study is a randomized controlled study. Fifty patients with pulmonary nodules scheduled for elective surgery in Zhongshan Hospital from January 2023 to September 2023 were randomly divided into the control group and the study group ( n=25 in each group). Patients in the study group carried out home pre-rehabilitation training guided by mobile App before operation, including breathing training, resistance exercise, nutritional intervention and psychological counseling; there was no intervention for patients in the control group while waiting for operation. The lung function, 6-min walking distance, psychological status, nutritional status, postoperative complications, catheterization time, hospitalization time, hospitalization expenses and complications were compared between the two groups before and after operation. Results:There were no significant differences in age, sex, BMI, heart rate, respiratory rate, blood pressure, pulmonary function, nutritional status, location of pulmonary nodules, anesthesia mode, operation modality and surgical pathology results between two groups(all P>0.05). Compared with the control group, the study group had significantly improved forced vital capacity at admission, 1d and 30d after surgery ( Z=-2.338, Z=-2.203, Z=-2.348, all P<0.05), forced expiratory volume in one second (FEV1) ( Z=-3.241, Z=-3.262, Z=-3.564, all P<0.05), and the ratio of forced vital capacity (FVC) to FEV1 ( t=-3.186, t=-2.904, and t=-4.304, all P<0.05), increased 6 min walking distance ( t=-4.466, t=-4.144, t=-4.907, all P<0.05), improved anxiety ( Z=-3.164, Z=-4.425, Z=-3.622, all P<0.05), decreased depression ( Z=-2.598, Z=-3.799, Z=-3.439, all P<0.05); and improved nutritional status at admission and 1d after surgery ( Z=-3.061, Z=-3.648, all P<0.05). The incidence of postoperative complications was decreased ( χ 2=4.878, P<0.05), the time of catheterization was decreased ( Z=-4.056, P<0.001), the length of hospital stay was shortened ( Z=-4.345, P<0.001), and the cost of hospitalization was reduced ( Z=-2.590, P<0.05) in the study group. Conclusion:The home-based online prehabilitation training can effectively improve the cardiopulmonary function and physical fitness of patients, reduce the stress of surgery, and contribute to rapid recovery after surgery; it also can shorten the length of postoperative hospital stay and reduce medical costs.

This experiment aims to study the taste-masking effects of different kinds of corrigent used individually and in combination on ibuprofen oral solution, in order to optimize the taste-masking formulation. Firstly, a wide range of corrigent and the mass fractions were extensively screened using electronic tongue technology. Subsequently, a combination of sensory evaluation, analytic hierarchy process (AHP)-fuzzy mathematics evaluation, and Box-Behnken experimental design were employed to comprehensively assess the taste-masking effects of different combinations of corrigent on ibuprofen oral solution, optimize the taste-masking formulation, and validate the results. The study received ethical approval from the Review Committee of the Beijing University of Chinese Medicine (ethical code: 2024BZYLL0102). The results showed that corrigent fractions and types were screened separately through single-factor experiments. Subsequently, a Box-Behnken response surface design combined with AHP and fuzzy mathematics evaluation was used to fit a functional model: Z = 688.310 11 - 3 023.722 22X₁ - 11.477 00X₂ + 62.721 67X₃ + 14.600 00X₁X₂ - 179.666 67X₁X₃ - 3.152 00X₂X₃ + 4 031.111 11X₁² + 0.525 28X₂² + 9.772 00X₃². This model is stable and reliable, determining the optimal taste-masking formulation to be 3.9 g·L^-1 of stevioside, 100 g L^-1 of xylitol, and 30 g L^-1 of methyl-β-cyclodextrin. The comprehensive score of the verification test is 88.14, with a relative RSD of 0.39%, indicating the feasibility of this model. This study achieved the improvement of the taste of ibuprofen oral solution through a combination of objective and subjective methods. Three types of corrigent were identified for enhancing drug taste, leading to the selection of the optimal taste-masking formulation for ibuprofen oral solution. This significantly enhanced the taste of the original formulation, improved patient compliance, and offered a new approach to mitigating the undesirable taste of formulations.

Objective To establish a renal cell co-culture system to simulate the renal barrier system,and to test its responsiveness to different glucose concentrations,and to investigate the regulatory effect of miR-29b-3p on osteopontin(OPN)/transforming growth factor β(TGF-β)pathway and the changes of this pathway under high glucose condition.Methods The three-cell co-culture system consisting of human renal podocytes,human glomerular mesangial cells and human renal tubular epithelial cells was established to test the cell viability and glucose consumption value at glucose concentrations of 5,8,12 and 16 mmol/L.The content of TGF-β and OPN in cell supernatant was measured.The recombinant plasmid and siRNA of OPN were transfected,and the expressions of TGF-β and OPN were detected by Q-PCR and Western blot.Results The mRNA expressions of OPN,TGF-β and miR-29b were significantly increased at 12 mmol/L glucose conditions.Western blot results showed that the protein expression of OPN increased in high glucose conditions,while the protein expression of TGF-β did not change significantly.After adding miR-29b-3p activator,the mRNA levels of OPN and TGF-β in the cell supernatant were significantly increased.After adding miR-29b-3p inhibitor,the mRNA levels of OPN and TGF-β in the cell supernatant were significantly decreased.Western blot results showed that compared with 5 mmol/L glucose,the protein expressions of OPN and TGF-β were increased by miR-29b-3p activator,and the protein expressions of OPN and TGF-β were decreased by miR-29b-3p inhibitor.After transfection with OPN recombinant plasmid,the content of TGF-β in the cell supernatant was significantly increased,and the mRNA expressions of OPN and TGF-β in the cells were significantly increased.After transfection with OPN siRNA,the content of TGF-β in the cell supernatant was decreased,and the expression of OPN mRNA in the cells was significantly decreased,but the expression of TGF-β mRNA was not significantly increased.Conclusion The renal cell co-culture system can mimic the complex renal environment in vivo.When induced by high glucose,cell proliferation is inhibited,glucose consumption is increased,and the content of TGF-β in the cell supernatant is increased,and miR-29b-3p has a regulatory effect on OPN/TGF-β signaling pathway in the co-culture system.

Objective:To evaluate the application of modular teaching program in the clinical competence training for community general practitioners.Methods:This was a cross-sectional study. A 140 teaching hour-modular training program was designed by the authors on the basis of the previous training experience and conducted in clinical competence training for 20 general practitioners (study group) from all 10 community health service centers in Shanghai Huangpu district between September 2022 and April 2023; and 26 general practitioners who planned to participate in the next batch of training courses were enrolled as the control group. The clinical competence assessment was conducted in May 2023, including knowledge level tests and clinical skill assessment. By comparing the scores of clinical competence assessment and satisfaction survey between two groups, the training effectiveness was evaluated.Results:Among the 46 general practitioners, 33 (71.7%) were female, aged 27 to 47 (37±6), working in general practice for 2 to 25 (9.4±5.5) years, 31 (67.4%) had participated in standardized residency training for general practice and 31 (67.4%) were contracted family doctors. The total score of 46 general practitioners was 69.7±9.6, and 68.9±8.6 for knowledge level and 69.8±10.6 for clinical skills. The total scores and single scores of trained participants were all significantly higher than those of untrained(all P<0.05). The multiple linear regression model showed that after controlling for gender, age, educational background, professional title, years of working in general practice, participation in standardized residency training, and status as a contracted family doctor, the trained participants had significantly higher scores in history collection, pediatric reception, indwelling gastric tube and abdominal puncture (all P<0.05). There was no statistically significant difference in the scores in knowledge level test, nervous system physical examination, SOAP medical record writing, and tutorium between the trained and untrained groups (all P>0.05). Among 20 general practitioners who completed training, the overall satisfaction score of the training program was 4.3±0.7 points, including 4.3±0.6 for the training course, 4.3±0.7 for teachers, and 4.3±0.7 for the organization and arrangement; 85% of the participants considered the training to be very helpful or helpful for improving their competence, and 90% considered the training to be very helpful or helpful for their future work. Conclusion:Modular teaching program is helpful to improve the clinical competence of community general practitioners, especially in clinical skills.

Objective: To examine the application value of 3D Slicer software assisted domestic frameless stereotactic robot in biopsy of intracranial lesions. Methods: A retrospective analysis was performed on 80 patients who admitted consecutively and underwent intracerebral lesions biopsy with the domestic frameless stereotactic robot at Department of Neurosurgery, Aerospace Central Hospital from January 2019 to December 2021. There were 36 males and 44 females, with a mean age of (38.5±18.0) years (range: 6 to 71 years). Before surgery only enhanced T1-weighted three-dimensional magnetization prepared gradient echo sequences and diffusion tensor imaging scans were performed. Self-reconstruction of intracranial lesions, cerebral cortex and blood vessels was carried out using 3D Slicer software system after the DICOM format imaging data of 80 patients were collected. These imaging data were merged to the workstation of the domestic frameless stereotactic robot for preoperative surgical planning and the surgical puncture path was designed to avoid blood vessels in the brain functional area, cerebral cortex and sulcus. Results: All frameless stereotactic biopsy were successfully performed. Postoperative pathological diagnosis included 50 cases of diffuse astrocytic and oligodendroglioma, 15 cases of lymphoma, 5 cases of metastatic tumors, 5 cases of inflammatory demyelinating disease, 2 cases of inflammatory granuloma, 1 case of hemangioma, 1 case of acute lymphoblastic leukemia intracranial invasion and 1 case of seminoma. The positive diagnosis rate was 100% (80/80). Postoperative imaging confirmed that the puncture path and target were accurately implemented according to the preoperative planning, and the target error was (1.32±0.44) mm (range: 0.55 to 1.99 mm). One case of puncture-related bleeding occurred at the target after surgery and improved after treatment. Conclusion: The three-dimensional multimodal images reconstructed by the 3D Slicer software before operation could help the surgeons make the preoperative planning and reduce the risk of stereotactic brain biopsy.
Male ; Female ; Humans ; Child ; Adolescent ; Young Adult ; Adult ; Middle Aged ; Aged ; Brain Neoplasms/pathology* ; Diffusion Tensor Imaging ; Retrospective Studies ; Robotics ; Biopsy ; Software ; Stereotaxic Techniques

In 2020, the prevalence of abnormal spinal curvature among 54 079 students in Shandong Province was 1.54%. The multivariate logistic regression model analysis showed that, compared with those in primary school, economically underdeveloped areas, and non-residential schools, students in middle and high schools, economically average areas, and residential schools had a higher risk of abnormal spinal curvature, with OR (95%CI) values of 2.029 (1.662-2.476), 2.746 (2.208-3.416), 2.237 (1.740-2.875) and 2.057 (1.705-2.483), respectively. Compared with those in economically underdeveloped areas, who were underweight, who had seat adjustments≤1 time per academic year, and who had physical education classes≤1 per week, students in economically developed areas, who were normal weight, overweight, and obese, who had seat adjustments≥2 times per academic year, and who had physical education classes 2-3 or≥4 per week, had a lower risk of abnormal spinal curvature, with OR (95%CI) values of 0.690 (0.521-0.915), 0.722 (0.546-0.955), 0.535 (0.389-0.735), 0.383 (0.274-0.535), 0.835 (0.711-0.980), 0.561 (0.474-0.663) and 0.491 (0.315-0.766), respectively.
Humans ; Risk Factors ; Prevalence ; Spinal Curvatures ; Schools ; Students

In 2020, the prevalence of abnormal spinal curvature among 54 079 students in Shandong Province was 1.54%. The multivariate logistic regression model analysis showed that, compared with those in primary school, economically underdeveloped areas, and non-residential schools, students in middle and high schools, economically average areas, and residential schools had a higher risk of abnormal spinal curvature, with OR (95%CI) values of 2.029 (1.662-2.476), 2.746 (2.208-3.416), 2.237 (1.740-2.875) and 2.057 (1.705-2.483), respectively. Compared with those in economically underdeveloped areas, who were underweight, who had seat adjustments≤1 time per academic year, and who had physical education classes≤1 per week, students in economically developed areas, who were normal weight, overweight, and obese, who had seat adjustments≥2 times per academic year, and who had physical education classes 2-3 or≥4 per week, had a lower risk of abnormal spinal curvature, with OR (95%CI) values of 0.690 (0.521-0.915), 0.722 (0.546-0.955), 0.535 (0.389-0.735), 0.383 (0.274-0.535), 0.835 (0.711-0.980), 0.561 (0.474-0.663) and 0.491 (0.315-0.766), respectively.
Humans ; Risk Factors ; Prevalence ; Spinal Curvatures ; Schools ; Students

Objective:To compare the breadth of condition coverage, accuracy of suggested conditions and appropriateness of urgency advice of the 8 symptom assessment mobile applications (APPs) available on the Chinese market.Methods:The APPs were assessed using 200 primary care vignettes and were measured against the vignettes′ standard. The primary outcome measures were proportion of conditions covered by an APP, proportion of vignettes with the correct primary diagnosis,and proportion of safe urgency advice.Results:For APPs assessed,condition-coverage was from 29.0%(58/200)to 99.5%(199/200), top-3 suggestion accuracy was from 8.5%(17/200) to 61.5%(123/200), the proportion of safe urgency advice was from 84.8%(167/197) to 99.5% (198/199).Conclusions:The APPs showed a wide range of coverage, safety performance and condition-suggestion accuracy. Symptom assessment APPs with good performance could be used by general practitioners as supporting tools. However, even symptom assessment APPs with excellent performance need to be further assessed in a real clinical environment.