ObjectiveTo obtain a construct chimeric antigen receptor macrophages （CAR-M） targeting disialoganglioside GD2 （GD2） and harboring the intracellular domain of macrophage-specific activating signaling adaptor protein DNAX-activating protein 12 （DAP12）. MethodsA CAR molecule targeting GD2 with the DAP12 intracellular domain as the signaling module ［Anti-GD2 CAR （DAP12）］ was designed， synthesized， and cloned into an adenoviral vector. This virus was used to infect PMA-differentiated THP-1 macrophages to construct green fluorescent protein （GFP）-labeled Anti-GD2 CAR THP-1 cells. CAR expression was confirmed using live-cell imaging. In vitro co-culture assays were performed to evaluate the phagocytic function of empty， Anti-GD2 CAR-M （CD3ζ）， and Anti-GD2 CAR-M （DAP12） groups against glioma cells with high （T98G） and low （U251） GD2 expression. ResultsThe CAR adenovirus efficiently transduced THP-1 macrophages， resulting in successful generation of Anti-GD2 CAR-M. Flow cytometry analysis following co-culture with glioma cells showed that compared to the empty group， the Anti-GD2 CAR-M （CD3ζ） group exhibited significantly enhanced phagocytosis （P<0.05）. Furthermore， compared to the Anti-GD2 CAR-M （CD3ζ） group， the Anti-GD2 CAR-M （DAP12） group demonstrated a further increase in phagocytic capacity specifically against GD2-high T98G cells （P<0.05）. ConclusionAnti-GD2 CAR-M （DAP12） cells are successfully established by transducing PMA-differentiated THP-1-M with recombinant Anti-GD2 CAR （DAP12） adenovirus. The engineered CAR-M can specifically target phagocytose glioma cells with high GD2 expression.

ObjectiveTo obtain stably transfected chimeric antigen receptor macrophages （CAR-M） targeting disialosylganglioside （GD2） based on the constitutive expression of myeloid differentiation factor 88 （MYD88）. MethodsAn Anti-GD2 CAR adenoviral vector incorporating the CD3ζ （cluster of differentiation 3 zeta chain）-MYD88 fusion construct was designed and constructed. THP-1-derived macrophages （THP-1M）， induced by phorbol 12-myristate 13-acetate （PMA） differentiation， were infected with this adenovirus to generate Anti-GD2 CAR THP-1 cells bearing a green fluorescent protein （GFP） tag. CAR expression was monitored using a live-cell imaging system. The phagocytic function of Empty CAR-M， Anti-GD2 CAR-M（CD3ζ）， and Anti-GD2 CAR-M（CD3ζ-MYD88） groups against glioma cells was evaluated in vitro through co-culture experiments， and their polarization status was also analyzed. ResultsCAR adenovirus successfully transfected THP-1 to obtain Anti-GD2 CAR-M. After co-culture with glioma cells， flow cytometry results showed that compared with the Empty CAR-M group and the Anti-GD2 CAR-M（CD3ζ） group， the Anti-GD2 CAR-M（CD3ζ-MYD88） group exhibited enhanced phagocytosis against T98G cells with high GD2 expression （P<0.05） and also showed upregulation of CD86 （P<0.05）. Compared with the Empty CAR-M group and the Anti-GD2 CAR-M（CD3ζ） group， the Anti-GD2 CAR-M（CD3ζ-MYD88）CAR-M group exhibited enhanced phagocytosis against T98G cells with high GD2 expression （P<0.05） and also showed upregulation of CD86 （P<0.05）， and the expression of inflammatory cytokines interleukin （IL）-6 and IL-12 was increased in this group （P<0.05）. ConclusionThis study successfully constructed Anti-GD2 CAR-M cells and verified their phagocytic activity， demonstrating that constitutive expression of MYD88 can promote inflammatory polarization of CAR-M. This provides a potential structural modification approach for the development of pro-inflammatory CAR-M.

Objective To study a method for determining the cumulative dose of low-energy neutrons ( < 100 keV) independently based on a CR-39 detector. Methods According to the theory of track etching kinetics, the differences in the tracks formed by low-energy neutrons or fast neutrons in a BN + CR-39 detector under broad-spectrum neutron irradiation were analyzed. A method was proposed to identify the tracks produced by low-energy neutrons under specific etching conditions while avoiding interference from fast neutron tracks. Results Experimental results demonstrated that the BN + CR-39 detector using TASTRAK PADC CR-39 track-detecting plastic could independently detect the tracks of low-energy neutrons when etched in a 6.25 mol/L NaOH solution for 1 h. The track density showed a good linear relationship with the ambient dose equivalent of low-energy neutrons, and the calibration coefficient was 0.0366 μSv·tr⁻¹·cm². Conclusion The BN + CR-39 detector proposed in this study demonstrated high sensitivity to low-energy neutrons. The track identification method can be used for effective and independent determination of the cumulative dose equivalent of low-energy neutrons.

This study aims to elucidate the role and mechanism of clematichinenoside AR(CAR) in protecting bone marrow mesenchymal stem cells(BMSCs) from hypoxia-induced apoptosis. BMSCs were isolated by the bone fragment method and identified by flow cytometry. Cells were cultured under normal conditions(37℃, 5% CO_2) and hypoxic conditions(37℃, 90% N_2, 5% CO_2) and treated with CAR. The BMSCs were classified into eight groups: control(normal conditions), CAR(normal conditions + CAR), hypoxia 24 h, hypoxia 24 h + CAR, hypoxia 48 h, hypoxia 48 h + CAR, hypoxia 72 h, and hypoxia 72 h + CAR. The cell counting kit-8(CCK-8) assay and terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL) were employed to measure cell proliferation and apoptosis, respectively. The number of mitochondria and mitochondrial membrane potential were measured by MitoTracker®Red CM-H2XRo staining and JC-1 staining, respectively. The level of reactive oxygen species(ROS) was measured with the DCFH-DA fluorescence probe. The protein levels of B-cell lymphoma-2 associated X protein(BAX), caspase-3, and optic atrophy 1(OPA1) were determined by Western blot. The results demonstrated that CAR significantly increased cell proliferation. Compared with the control group, the hypoxia groups showed increased apoptosis rates, reduced mitochondria, elevated ROS levels, decreased mitochondrial membrane potential, upregulated expression of BAX and caspase-3, and downregulated expression of OPA1. In comparison to the corresponding hypoxia groups, CAR intervention significantly decreased the apoptosis rate, increased mitochondria, reduced ROS levels, elevated mitochondrial membrane potential, downregulated the expression of BAX and caspase-3, and upregulated the expression of OPA1. Therefore, it can be concluded that CAR may exert an anti-apoptotic effect on BMSCs under hypoxic conditions by regulating OPA1 to maintain mitochondrial homeostasis.
Mesenchymal Stem Cells/metabolism* ; Apoptosis/drug effects* ; Mitochondria/metabolism* ; Animals ; Rats ; Cell Hypoxia/drug effects* ; Homeostasis/drug effects* ; Reactive Oxygen Species/metabolism* ; Rats, Sprague-Dawley ; Membrane Potential, Mitochondrial/drug effects* ; Saponins/pharmacology* ; Caspase 3/genetics* ; Male ; bcl-2-Associated X Protein/genetics* ; Bone Marrow Cells/metabolism* ; Cell Proliferation/drug effects* ; Protective Agents/pharmacology* ; Cells, Cultured

Objective:To explore the influencing factors of dietary habits on the progression from prediabetes to type 2 diabetes mellitus(T2DM)in elderly individuals undergoing health check-ups.Methods:In the cross-sectional study, we enrolled individuals aged 60-70 years with fasting plasma glucose (FPG)≥6.0 mmol/L who underwent health examinations at the Health Management Medical Center of Wenjiang District People's Hospital in Chengdu from 2019 to 2022.Demographic characteristics, dietary habit questionnaires, and FPG values were collected.Unconditional binary logistic regression analysis was used to identify factors influencing the natural progression from prediabetes to T2DM.A nomogram prediction model was established based on logistic regression results, and its predictive performance was evaluated by calculating the C-statistics and drawing a calibration curve.Results:A total of 13 681 elderly participants with FPG ≥6.0 mmol/L were included, comprising 4 306(31.5%)prediabetes cases(FPG 6.0-7.0 mmol/L), aged(63.54±16.49)years and 9 375(68.5%)T2DM cases(FPG>7.0 mmol/L), aged(63.09±16.21)years.Unconditional binary logistic regression analysis showed that frequent breakfast( OR=0.777, 95% CI: 0.696-0.868, P<0.001), dietary preference for light diet( OR=0.781, 95% CI: 0.710-0.858, P<0.001), salty taste( OR=0.571, 95% CI: 0.504-0.648, P<0.001), raw food( OR=0.327, 95% CI: 0.224-0.478, P<0.001)and spicy taste( OR=0.124, 95% CI: 0.112-0.137, P<0.001)were the protective factors for the conversion of prediabetes to the T2DM stage in the elderly physical examination population.While fast eating rate( OR=4.327, 95% CI: 3.978-4.772, P<0.001), dietary preference for sweets( OR=5.168, 95% CI: 4.703-5.678, P<0.001), and high-fat diet( OR=1.401, 95% CI: 1.275-1.539, P<0.001)were risk factors for conversion of prediabetes to T2DM stage.C-statistic of the Nomogram prediction model was 0.781; the goodness-of-fit test of the calibration curve was χ2=11.258, P=0.188, and the model predicted well. Conclusions:Regular breakfast, light diet, and dietary preferences for salty, raw, and spicy foods were protective factors for the transition from prediabetes to T2DM stage, whereas rapid eating rate, preference for sweets, and high-fat diets were risk factors for the transition from prediabetes to T2DM stage in the medical examination population.The constructed risk prediction model helped to find out the magnitude of the risk of T2DM in an individual, which increases the evidence for the transition from prediabetes to T2DM stage prevention evidence.

Objective:To explore the influencing factors of dietary habits on the progression from prediabetes to type 2 diabetes mellitus(T2DM)in elderly individuals undergoing health check-ups.Methods:In the cross-sectional study, we enrolled individuals aged 60-70 years with fasting plasma glucose (FPG)≥6.0 mmol/L who underwent health examinations at the Health Management Medical Center of Wenjiang District People's Hospital in Chengdu from 2019 to 2022.Demographic characteristics, dietary habit questionnaires, and FPG values were collected.Unconditional binary logistic regression analysis was used to identify factors influencing the natural progression from prediabetes to T2DM.A nomogram prediction model was established based on logistic regression results, and its predictive performance was evaluated by calculating the C-statistics and drawing a calibration curve.Results:A total of 13 681 elderly participants with FPG ≥6.0 mmol/L were included, comprising 4 306(31.5%)prediabetes cases(FPG 6.0-7.0 mmol/L), aged(63.54±16.49)years and 9 375(68.5%)T2DM cases(FPG>7.0 mmol/L), aged(63.09±16.21)years.Unconditional binary logistic regression analysis showed that frequent breakfast( OR=0.777, 95% CI: 0.696-0.868, P<0.001), dietary preference for light diet( OR=0.781, 95% CI: 0.710-0.858, P<0.001), salty taste( OR=0.571, 95% CI: 0.504-0.648, P<0.001), raw food( OR=0.327, 95% CI: 0.224-0.478, P<0.001)and spicy taste( OR=0.124, 95% CI: 0.112-0.137, P<0.001)were the protective factors for the conversion of prediabetes to the T2DM stage in the elderly physical examination population.While fast eating rate( OR=4.327, 95% CI: 3.978-4.772, P<0.001), dietary preference for sweets( OR=5.168, 95% CI: 4.703-5.678, P<0.001), and high-fat diet( OR=1.401, 95% CI: 1.275-1.539, P<0.001)were risk factors for conversion of prediabetes to T2DM stage.C-statistic of the Nomogram prediction model was 0.781; the goodness-of-fit test of the calibration curve was χ2=11.258, P=0.188, and the model predicted well. Conclusions:Regular breakfast, light diet, and dietary preferences for salty, raw, and spicy foods were protective factors for the transition from prediabetes to T2DM stage, whereas rapid eating rate, preference for sweets, and high-fat diets were risk factors for the transition from prediabetes to T2DM stage in the medical examination population.The constructed risk prediction model helped to find out the magnitude of the risk of T2DM in an individual, which increases the evidence for the transition from prediabetes to T2DM stage prevention evidence.

In order to investigate the chemical constituents of glycosides in Piper sintenense Hatusima, column chromatographic techniques such as silica gel, ODS, MCI GEL CHP20P, Sephadex LH-20, and semi-preparative high performance liquid chromatography were used to afford nine glycosides from the n-butanol part of the 95% ethanol extract of Piper sintenense Hatusima. Based on the physicochemical properties and NMR data, the above compounds were identified as (2S)-2-hydroxy-1-(4-hydroxy-3-methoxyphenyl)-1-propanone-2-O-β-D-glucopyranoside (1), 2-phenylethyl β-D-glucopyranoside (2), benzyl α-L-arabinopyranosyl-(1''→6')-β-D-glucopyranoside (3), benzyl β-D-xylopyanosyl-(1''→6')-β-D-glucopyranoside (4), phenethyl β-D-apiofuranosyl-(1''→ 2')-β-D-glucopyranoside(5), salidroside (6), phenethanol β-D-xylopyanosyl-(1''→6')-β-D-glucopyranoside (7), (Z)-hexenyl-O-α-L-arabinopyranosyl-(1''→6')-O-β-D-glucopyranoside (8), (Z)-hexenyl-O-β-D-xylopyanosyl-(1''→6')-O-β-D-glucopyranoside (9). Compound 1 was identified as a new compound, and compounds 3-9 were isolated from the genus Piper for the first time.

Liver size is regulated by circadian clock and exhibits a diurnal rhythm. Pregnane X receptor (PXR) and peroxisome proliferator-activated receptor α (PPARα), members of nuclear receptor superfamily, are important regulators of liver size. We previously demonstrated that mPXR agonist pregnenolone 16α-carbonitrile (PCN) and mPPARα agonist pirinixic acid (WY-14643) promoted liver enlargement and liver regeneration after partial hepatectomy. However, whether PXR or PPARα activation-induced liver enlargement exhibits diurnal rhythm with normal liver diurnal oscillations remains unclear. The aim of this study was to investigate the diurnal rhythm of PXR or PPARα activation-induced liver enlargement. Male C57BL/6 mice were intraperitoneally injected with corn oil, PCN or WY-14643 for three days, and liver samples were weighed and collected at various time points for analysis. The animal experiment protocol was reviewed and approved by Institutional Animal Care and Use Committee of Sun Yat-sen University (approval No. SYSU-IACUC-2023-001613, SYSU-IACUC-2023-001783). The results showed that PXR or PPARα activation at various time points significantly induced liver enlargement, and liver size maintained normal diurnal oscillations during PXR or PPARα activation-induced hepatomegaly, without significant effects on the expression of core clock genes. This study reveals PXR or PPARα activation-induced liver enlargement exhibits diurnal rhythm with normal liver diurnal oscillations, and provides novel data for nuclear receptor-induced liver enlargement and liver diurnal rhythm.

Objective:To explore the rules of Traditional Chinese Medicine (TCM) prescriptions of gastroesophageal reflux disease based on Ancient and Modern Medical Records Cloud Platform.Method:The relevant medical cases from ancient medical case database, modern medical case database, shared medical case database and famous doctors' medical case database in Ancient and Modern Medical Records Cloud Platform (V2.3.8) were selected, and frequency analysis, attribute analysis, association analysis, cluster analysis and complex network analysis were performed on the herbs.Results:A total of 107 medical records were obtained, including 225 TCMs. The core medicines were Radix et Rhizoma Glycyrrhizae, Pericarpium Citri Reticulatae, Rhizoma Coptidis, Poria, and Fructus Evodiae. The drug property was mainly cold and warm, and the herbal tastes bitter and pungent. The meridian tropism of drugs mainly manifested in the spleen and stomach meridians. The core herbal pairs were Radix et Rhizoma Glycyrrhizae and Pericarpium Citri Reticulatae. The core prescription consisted of 17 herbs including Radix Glycyrrhizae, Pericarpium Citri Reticulatae, Rhizoma Coptidis, Fructus Evodiae, Poria, Endoconcha Sepiae, Herba Taraxaci, Fructus Aurantii, Radix Paeoniae Alba, Radix Bupleuri, Jiang Banxia, Rhizoma Cyperi, Radix Aucklandiae, Caulis Bambusae In Taenia, Fructus Aurantii Immaturus, Fructus Amomi, and Rhizoma Atractylodis Macrocephalae. Conclusions:Chinese medicine treatment of gastroesophageal reflux disease is mainly based on Chaihu Shugan Powder, Zuojin Pill, and Wendan Decoction. Moreover, we need to combine with clinical symptoms to add or subtract herbs.

Objective: To analyze and compare therapy responses, outcomes, and incidence of severe hematologic adverse events of flumatinib and imatinib in patients newly diagnosed with chronic phase chronic myeloid leukemia (CML) . Methods: Data of patients with chronic phase CML diagnosed between January 2006 and November 2022 from 76 centers, aged ≥18 years, and received initial flumatinib or imatinib therapy within 6 months after diagnosis in China were retrospectively interrogated. Propensity score matching (PSM) analysis was performed to reduce the bias of the initial TKI selection, and the therapy responses and outcomes of patients receiving initial flumatinib or imatinib therapy were compared. Results: A total of 4 833 adult patients with CML receiving initial imatinib (n=4 380) or flumatinib (n=453) therapy were included in the study. In the imatinib cohort, the median follow-up time was 54 [interquartile range (IQR), 31-85] months, and the 7-year cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) were 95.2%, 88.4%, 78.3%, and 63.0%, respectively. The 7-year FFS, PFS, and OS rates were 71.8%, 93.0%, and 96.9%, respectively. With the median follow-up of 18 (IQR, 13-25) months in the flumatinib cohort, the 2-year cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) were 95.4%, 86.5%, 58.4%, and 46.6%, respectively. The 2-year FFS, PFS, and OS rates were 80.1%, 95.0%, and 99.5%, respectively. The PSM analysis indicated that patients receiving initial flumatinib therapy had significantly higher cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) and higher probabilities of FFS than those receiving the initial imatinib therapy (all P<0.001), whereas the PFS (P=0.230) and OS (P=0.268) were comparable between the two cohorts. The incidence of severe hematologic adverse events (grade≥Ⅲ) was comparable in the two cohorts. Conclusion: Patients receiving initial flumatinib therapy had higher cumulative incidences of therapy responses and higher probability of FFS than those receiving initial imatinib therapy, whereas the incidence of severe hematologic adverse events was comparable between the two cohorts.
Adult ; Humans ; Adolescent ; Imatinib Mesylate/adverse effects* ; Incidence ; Antineoplastic Agents/adverse effects* ; Retrospective Studies ; Pyrimidines/adverse effects* ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy* ; Treatment Outcome ; Benzamides/adverse effects* ; Leukemia, Myeloid, Chronic-Phase/drug therapy* ; Aminopyridines/therapeutic use* ; Protein Kinase Inhibitors/therapeutic use*