OBJECTIVE To optimize the extraction process of flavonoids from Potentilla discolor .METHODS Deep eutectic solvents （DESs） were used as the extraction solvent to prepare test sample solutions. The contents of rutin， quercitrin， and kaempferol were determined by ultra-performance liquid chromatography. The analytic hierarchy process combined with the criteria importance through intercriteria correlation weighting method was employed to assign comprehensive weights to the contents of the three flavonoids， and calculate the comprehensive score. The optimal DESs were screened based on the comprehensive score.Mean while， using the comprehensive score as the response variable and the liquid-to-solid ratio， ultrasonic time， and urltrasonic temperature as the influencing factors， the extraction technology of flavonoids from P.discolor was optimized by Box-Behnken response surface methodology and subsequently validated.RESULTS The comprehensive weight coefficients of rutin， quercitrin， and kaempferol were 0.646 7， 0.207 5， and 0.145 8， respectively. The optimal DESs were choline chloride-lactic acid （molar ratio 1∶4） with a water content of 40%. The optimal extraction conditions were determined as follows： liquid-to-solid ratio of 25∶1， ultrasonic time of 40 min， and ultrasonic temperature of 50 ℃. The validation results showed that the average comprehensive score of three batches of samples was 99.28， with an RSD of 0.54% （ n ＝3）.CONCLUSIONS The optimal extraction technology of flavonoids from P. discolor based on DESs obtained in this study was stable and feasible.

Objective To explore the method for establishing a pig left lung orthotopic transplantation model. Methods A total of 4 male Yorkshire pigs weighing (40.0±2.5) kg underwent surgical procedures, including anesthesia, tracheal intubation, donor lung procurement, and recipient lung transplantation. Histological morphology and blood gas analysis of the transplanted lungs were assessed 2 hours after reperfusion to evaluate the histological changes and function of the transplanted lungs. Results The pigs underwent left lung in situ transplantation, effectively simulating conditions relevant to human lung transplantation. Two hours after the transplantation, arterial blood gas analysis showed that arterial partial pressure of oxygen was 155.4-178.6 mm Hg, arterial partial pressure of carbon dioxide was 53.1-62.4 mm Hg, and the oxygenation index was 310.8-357.2 mm Hg. Hematoxylin and eosin staining indicated a low degree of pulmonary edema and minimal cellular infiltration. Conclusion The pig left lung orthotopic transplantation model possesses strong operability and stability. Researchers can replicate this model according to the described methods and further conduct basic research and explore clinical translational applications.

Toxoplasmosis, a zoonotic disease caused by the obligate intracellular parasite Toxoplasma gondii, is widely prevalent across the world. The life cycle of T. gondii involves definitive hosts (cats and felines), intermediate hosts (humans, livestock, and wild animals), and the environment (such as soil and water). The One Health concept brings together the health of humans, animals, and the environment, with aims to pursue the optimal health outcomes through coordinated management. Management of toxoplasmosis from a One Health perspective may provide key insights into formulation of interventions for toxoplasmosis control. This article reviews the complex biological characteristics and life cycle of T. gondii, as well as the importance of felines as definitive hosts in the transmission of toxoplasmosis, analyzes the epidemiology, ecological impact and economic burden of toxoplasmosis, and discusses the bottlenecks in diagnostic and monitoring systems, environmental management technologies, and One Health interdisciplinary collaboration. In addition, toxoplasmosis control strategies are proposed based on the One Health concept, covering public health interventions, management of animal sources, multi-sector collaboration, and development of novel vaccines. These strategies provide insights into interruption of toxoplasmosis transmission and maintenance of the global public health security.

ObjectiveColorectal cancer (CRC) is a highly heterogeneous malignancy, and traditional Chinese medicine (TCM) syndrome differentiation is widely used in its clinical management. However, the biological basis underlying different TCM syndromes remains insufficiently understood. This study aimed to investigate the proteomic differences between qi-blood deficiency syndrome (QBDS) and damp-heat syndrome (DHS) in CRC patients and to establish a “syndrome-differential protein-herbal component” network for exploring potential molecular mechanisms and therapeutic targets associated with syndrome-specific treatment strategies. MethodsTumor tissue samples were collected from 10 patients with pathologically confirmed CRC, including 5 patients diagnosed with QBDS and 5 with DHS according to standardized TCM syndrome differentiation criteria. Tandem mass tag (TMT)-based quantitative proteomics was employed to identify differentially expressed proteins (DEPs) between the two syndrome groups. Functional enrichment analyses, including Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, were performed to characterize the biological functions and signaling pathways associated with the identified DEPs. Least absolute shrinkage and selection operator (LASSO) regression analysis was further applied to screen key syndrome-related proteins and construct a core protein prediction model. Subsequently, molecular docking was conducted to evaluate the binding affinities between representative herbal compounds, astragaloside IV and berberine, and the identified core proteins. Finally, cell counting kit-8 (CCK-8) assays were performed in HCT 116 and HT-29 colorectal cancer cell lines to validate the anti-proliferative activities of these compounds in vitro. ResultsA total of 44 DEPs were identified between QBDS and DHS tissues, including 21 upregulated and 23 downregulated proteins in the QBDS group relative to the DHS group. Functional enrichment analyses revealed that these proteins were primarily associated with antiviral immune responses, regulation of inflammatory signaling, NF‑κB signaling pathways, cholesterol metabolism, and other biological processes relevant to tumor progression and host immune regulation. LASSO regression analysis identified eight core proteins with potential syndrome-specific significance. Among them, ST7, GPCPD1, and PODXL were closely associated with QBDS, whereas FAT1, GRB7, KRT7, MAFF, and MMP8 were associated with DHS. Molecular docking demonstrated favorable interactions between astragaloside IV and QBDS-related proteins, with binding energies lower than -5 kcal/mol, indicating stable binding activity. Berberine exhibited even lower binding energies and stronger binding specificity toward DHS-related core proteins. In vitro experiments further confirmed that both compounds inhibited CRC cell proliferation, while berberine displayed significantly greater anti-proliferative efficacy. The lowest 48 h IC₅₀ value of berberine reached 15.21 mg/L in CRC cells, indicating a stronger growth-inhibitory effect than astragaloside IV. ConclusionCRC patients with QBDS and DHS exhibit distinct proteomic characteristics, suggesting that different TCM syndromes possess unique molecular signatures. The identified core proteins may serve as potential biomarkers for syndrome differentiation, while astragaloside IV and berberine may exert therapeutic effects through targeting syndrome-specific molecular networks. These findings provide preliminary proteomic evidence supporting syndrome-guided precision integrative medicine in CRC and offer new insights into the biological basis of TCM syndrome differentiation and individualized therapeutic strategies.

This paper summarizes professor LIU Zhibin's clinical experience in treating Parkinson's disease with festinating gait by acupuncture combined with tuina based on the theory of the eight hollow spaces. Professor LIU holds that the disease involve the channel sinews and joints， and that the key pathogenesis lies in dysfunction of the eight hollow spaces， impairment of mechanical pivots and contraction of channel sinews. Clinically， these manifestations are characterized by impaired function of the hip and popliteal regions with dysregulation of gait initiation and termination， as well as impaired function of the axillary and elbow regions with discoordination of postural balance and movement. The therapeutic principle is regulating the spirit， unblocking the collaterals， and facilitating the eight hollow spaces. Acupuncture focuses on awakening the brain， regulating the spirit， unblocking the eight hollow spaces and dredging the sinew channels， whereas tuina focuses on releasing sinew knots around the hip and popliteal regions and restoring the movement of mechanical pivots. Particular emphasis is placed on the sequential application of acupuncture and tuina and the integration of static and dynamic approaches， with physical movement used to facilitate regulation of the spirit and gait training employed to consolidate therapeutic effects. This approach provides a new perspective for the treatment of festinating gait in Parkinson's disease with traditional Chinese medicine.

BACKGROUND:Aging of human body may be due to the senescence and depletion of various stem cells in the body.Bone marrow mesenchymal stromal cells have important physiological functions and have shown certain therapeutic effects on various diseases.It is of great significance to study the senescence and mechanism of bone marrow mesenchymal stromal cells.OBJECTIVE:To investigate whether human bone marrow mesenchymal stromal cells exhibit senescence phenotypes with increasing donor age,and further determine whether endogenous retrovirus drives the senescence of bone marrow mesenchymal stromal cells,offering a novel reference for the investigation of stem cell senescence mechanism.METHODS:The senescence of bone marrow mesenchymal stromal cells at different ages was characterized by flow cytometry,β-galactosidase staining,qPCR,western blotting,and EdU fluorescence imaging.Bone marrow mesenchymal stromal cells and cell culture supernatant were collected from donors of different ages.The content of human endogenous retrovirus was detected by qPCR.Furthermore,highly sensitive droplet digital PCR was used to detect the expression of endogenous retrovirus-like particles in the cell culture supernatant.The content of endogenous retrovirus in bone marrow plasma samples of different ages was detected by ELISA.RESULTS AND CONCLUSION:Bone marrow mesenchymal stromal cells exhibited obvious senescence with increasing age,including significant morphological changes,increased proportion of β-galactosidase positive cells,increased expression of senescence markers P16 and P21 protein,decreased expression of LMNB1 protein,and reduced cell proliferation ability.There was no significant difference in the content of endogenous retrovirus in bone marrow mesenchymal stromal cells at different ages,almost no endogenous retrovirus-like particles in the cell culture supernatant.There was no significant difference in endogenous retrovirus-like particles detected in bone marrow plasma samples at different ages.These findings indicate that human bone marrow mesenchymal stromal cells have normal physiological senescence with increasing age,but the mechanism of senescence is not mediated by abnormal activation of endogenous retroviruses,which may have a more complex driving mechanism.

BACKGROUND:Aging of human body may be due to the senescence and depletion of various stem cells in the body.Bone marrow mesenchymal stromal cells have important physiological functions and have shown certain therapeutic effects on various diseases.It is of great significance to study the senescence and mechanism of bone marrow mesenchymal stromal cells.OBJECTIVE:To investigate whether human bone marrow mesenchymal stromal cells exhibit senescence phenotypes with increasing donor age,and further determine whether endogenous retrovirus drives the senescence of bone marrow mesenchymal stromal cells,offering a novel reference for the investigation of stem cell senescence mechanism.METHODS:The senescence of bone marrow mesenchymal stromal cells at different ages was characterized by flow cytometry,β-galactosidase staining,qPCR,western blotting,and EdU fluorescence imaging.Bone marrow mesenchymal stromal cells and cell culture supernatant were collected from donors of different ages.The content of human endogenous retrovirus was detected by qPCR.Furthermore,highly sensitive droplet digital PCR was used to detect the expression of endogenous retrovirus-like particles in the cell culture supernatant.The content of endogenous retrovirus in bone marrow plasma samples of different ages was detected by ELISA.RESULTS AND CONCLUSION:Bone marrow mesenchymal stromal cells exhibited obvious senescence with increasing age,including significant morphological changes,increased proportion of β-galactosidase positive cells,increased expression of senescence markers P16 and P21 protein,decreased expression of LMNB1 protein,and reduced cell proliferation ability.There was no significant difference in the content of endogenous retrovirus in bone marrow mesenchymal stromal cells at different ages,almost no endogenous retrovirus-like particles in the cell culture supernatant.There was no significant difference in endogenous retrovirus-like particles detected in bone marrow plasma samples at different ages.These findings indicate that human bone marrow mesenchymal stromal cells have normal physiological senescence with increasing age,but the mechanism of senescence is not mediated by abnormal activation of endogenous retroviruses,which may have a more complex driving mechanism.

BACKGROUND: The contraction behaviors of cardiomyocytes (CMs), especially contraction synchrony, are crucial factors reflecting their maturity and response to drugs. A wider field of view helps to observe more pronounced synchrony differences, but the accompanied greater computational load, requiring more computing power or longer computational time. METHODS: We proposed a method that directly correlates variations in optical field brightness with cardiac tissue contraction status (CVB method), based on principles from physics and photometry, for rapid video analysis in wide field of view to obtain contraction parameters, such as period and contraction propagation direction and speed. RESULTS: Through video analysis of human induced pluripotent stem cell (hiPSC)-derived CMs labeled with green fluorescent protein (GFP) cultured on aligned and random nanofiber scaffolds, the CVB method was demonstrated to obtain contraction parameters and quantify the direction and speed of contraction within regions of interest (ROIs) in wide field of view. The CVB method required less computation time compared to one of the contour tracking methods, the LucasKanade (LK) optical flow method, and provided better stability and accuracy in the results. CONCLUSION This method has a smaller computational load, is less affected by motion blur and out-of-focus conditions, and provides a potential tool for accurate and rapid analysis of cardiac tissue contraction synchrony in wide field of view without the need for more powerful hardware.

BACKGROUND: The contraction behaviors of cardiomyocytes (CMs), especially contraction synchrony, are crucial factors reflecting their maturity and response to drugs. A wider field of view helps to observe more pronounced synchrony differences, but the accompanied greater computational load, requiring more computing power or longer computational time. METHODS: We proposed a method that directly correlates variations in optical field brightness with cardiac tissue contraction status (CVB method), based on principles from physics and photometry, for rapid video analysis in wide field of view to obtain contraction parameters, such as period and contraction propagation direction and speed. RESULTS: Through video analysis of human induced pluripotent stem cell (hiPSC)-derived CMs labeled with green fluorescent protein (GFP) cultured on aligned and random nanofiber scaffolds, the CVB method was demonstrated to obtain contraction parameters and quantify the direction and speed of contraction within regions of interest (ROIs) in wide field of view. The CVB method required less computation time compared to one of the contour tracking methods, the LucasKanade (LK) optical flow method, and provided better stability and accuracy in the results. CONCLUSION This method has a smaller computational load, is less affected by motion blur and out-of-focus conditions, and provides a potential tool for accurate and rapid analysis of cardiac tissue contraction synchrony in wide field of view without the need for more powerful hardware.

Objective To systematically evaluate the risk prediction models for anastomotic leakage (AL) in patients with esophageal cancer after surgery. Methods A computer-based search of PubMed, EMbase, Web of Science, Cochrane Library, Chinese Medical Journal Full-text Database, VIP, Wanfang, SinoMed and CNKI was conducted to collect studies on postoperative AL risk prediction model for esophageal cancer from their inception to October 1st, 2023. PROBAST tool was employed to evaluate the bias risk and applicability of the model, and Stata 15 software was utilized for meta-analysis. Results A total of 19 literatures were included covering 25 AL risk prediction models and 7373 patients. The area under the receiver operating characteristic curve (AUC) was 0.670-0.960. Among them, 23 prediction models had a good prediction performance (AUC>0.7); 13 models were tested for calibration of the model; 1 model was externally validated, and 10 models were internally validated. Meta-analysis showed that hypoproteinemia (OR=9.362), postoperative pulmonary complications (OR=7.427), poor incision healing (OR=5.330), anastomosis type (OR=2.965), preoperative history of thoracoabdominal surgery (OR=3.181), preoperative diabetes mellitus (OR=2.445), preoperative cardiovascular disease (OR=3.260), preoperative neoadjuvant therapy (OR=2.977), preoperative respiratory disease (OR=4.744), surgery method (OR=4.312), American Society of Anesthesiologists score (OR=2.424) were predictors for AL after esophageal cancer surgery. Conclusion At present, the prediction model of AL risk in patients with esophageal cancer after surgery is in the development stage, and the overall research quality needs to be improved.