1.Three-dimensional Electrical Impedance Tomography for Monitoring Gastric Hemorrhage
Zi-Han ZHAO ; Bo SUN ; Jing-Shi HUANG ; Zhi-Wei LI ; Yang WU ; Nan LI ; Jia-Feng YAO ; Tong ZHAO
Progress in Biochemistry and Biophysics 2026;53(4):1062-1075
ObjectiveGastric hemorrhage is one of the most common and life-threatening emergencies of the upper digestive tract. Early identification and continuous monitoring are essential for reducing rebleeding rates and mortality, particularly within the critical early hours after onset. Although endoscopy and radiological imaging can accurately localize bleeding sites, these approaches are invasive, resource-intensive, and unsuitable for continuous bedside monitoring. Electrical impedance tomography (EIT), as a noninvasive and radiation-free functional imaging technique, offers real-time visualization of conductivity distribution and has the potential for detecting intragastric bleeding based on the electrical contrast between blood and surrounding gastric tissues. In this study, a three-dimensional gastric EIT (3D-gEIT) framework is proposed to achieve noninvasive, real-time, and dynamic monitoring of gastric hemorrhage, with emphasis on spatial localization and quantitative volume assessment. MethodsA three-dimensional upper-abdominal simulation model incorporating the stomach, gastric wall, gastric contents, and surrounding tissues was established. Three electrode configurations, namely the dual layer ring, the four layer staggered ring, and the opposed dual plane array, were designed and systematically compared to evaluate their influence on depth sensitivity and spatial resolution. Based on the Tikhonov-Noser hybrid regularization scheme, a region-clustering constraint was introduced to develop the TK-Noser-RCC algorithm. This approach aggregates spatially adjacent elements with similar conductivity variations, thereby enhancing structural continuity and suppressing isolated noise artifacts. To validate the proposed framework, an upper-abdominal physical phantom was constructed using agar to simulate background tissue conductivity. Hemispherical high-conductivity inclusions with volumes ranging from 10 ml to 50 ml were attached to the inner gastric wall to mimic localized bleeding under different gastric filling states. Boundary voltages were acquired under a 120 kHz excitation current and reconstructed using the TK-Noser-RCC algorithm. Furthermore, an in vivo animal experiment was performed using a porcine model with adult-scale abdominal dimensions. A total of 100 ml of autologous blood was injected incrementally into the stomach to simulate progressive gastric hemorrhage, and time-difference EIT reconstruction was conducted at each injection stage to assess the dynamic system response under physiological conditions. ResultsSimulation results demonstrated that the opposed dual-plane electrode array achieved superior depth sensitivity distribution and spatial resolution. For a 40 ml hemorrhage model, the average ICC and SSIM improved by 55.9% and 38.8% compared with the dual-layer ring configuration, and by 64.0% and 39.5% compared with the four-layer staggered configuration. The proposed region-clustering constraint significantly enhanced reconstruction stability. Under added Gaussian noise of 40 dB and 30 dB, ICC values remained approximately 0.85, indicating effective artifact suppression and preservation of boundary integrity. In physical phantom experiments, reconstructed hemorrhage volumes increased approximately linearly with the preset hemispherical volumes, and the reconstructed high-conductivity regions closely matched the actual bleeding locations. Both empty-stomach and full-stomach conditions were evaluated, demonstrating that the opposed dual-plane configuration maintained stable imaging performance across varying gastric contents. In the animal experiment, reconstructed low-impedance regions expanded progressively with increasing injected blood volume. The spatial localization of the hemorrhage remained stable throughout the procedure, and no significant artifacts were observed. Quantitative analysis showed that reconstructed volume and average conductivity variation exhibited an approximately linear growth trend with injected blood volume, confirming the sensitivity of the system to dynamic intragastric conductivity changes. ConclusionThe proposed 3D-gEIT framework enables quantitative reconstruction of gastric hemorrhage volume and spatial distribution with improved depth sensitivity, structural continuity, and noise robustness compared with conventional EIT approaches. By integrating optimized electrode configuration and a region-clustering-constrained reconstruction algorithm, the system provides stable dynamic monitoring under both controlled phantom conditions and in vivo physiological environments. This method offers a noninvasive, real-time, and low-cost imaging strategy for early diagnosis, postoperative monitoring, and bedside surveillance of gastric bleeding.
2.Effect of Acupuncture at Neiguan (PC6) on Improving Autism by Promoting Myelination Through The METTL14/m⁶A/PTEN Axis Based on “Xuanfu-Suiqiao” Theory
Wei-Li DANG ; Lü-Yuan LIANG ; Yu-Xin LI ; Zhi-Yao LI ; Sai-Dan LIU ; Jia-Lei CAO ; Rong-Ze MA ; Yun-Kai WANG ; Xiao-Qing YANG ; Bing-Qi WEI ; Bing-Xiang MA
Progress in Biochemistry and Biophysics 2026;53(5):1165-1177
ObjectiveTo clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology. MethodsThe ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m6A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture. ResultsBehaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m6A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01). ConclusionMETTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m6A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of “opening Xuanfu and dredging myelin”.
3.Effect of Acupuncture at Neiguan (PC6) on Improving Autism by Promoting Myelination Through The METTL14/m⁶A/PTEN Axis Based on “Xuanfu-Suiqiao” Theory
Wei-Li DANG ; Lü-Yuan LIANG ; Yu-Xin LI ; Zhi-Yao LI ; Sai-Dan LIU ; Jia-Lei CAO ; Rong-Ze MA ; Yun-Kai WANG ; Xiao-Qing YANG ; Bing-Qi WEI ; Bing-Xiang MA
Progress in Biochemistry and Biophysics 2026;53(5):1165-1177
ObjectiveTo clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology. MethodsThe ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m6A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture. ResultsBehaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m6A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01). ConclusionMETTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m6A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of “opening Xuanfu and dredging myelin”.
4.Based on Experimental Verification, Mechanism of Euphorbia humifusa in Treatment of Acute Kidney Injury was Explored
Lijuan ZHANG ; Xuehai JIA ; Yaping GUO ; Shunying LI ; Lu YANG ; Dahong YAO ; Ke ZHANG ; Hangyu WANG ; Jinhui WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):166-176
ObjectiveTo explore the efficacy and mechanism of Euphorbia humifusa on acute kidney injury (AKI) based on network pharmacology, molecular docking and experimental verification. MethodsThe active components and targets of E. humifusa were retrieved from TCMSP and SwissTargetPrediction database, and the AKI targets were screened by GeneCards and Online Mendelian Inheritance in Man(OMIM) databases. The drug targets and disease targets were intersected to construct a protein-protein interaction network, and the intersection targets were subjected to gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis. Discover Studio software was used to verify the molecular docking of key components and core targets. Gentamicin (GM) was used to induce AKI rat model. Control group, model group, verapamil (16 mg·kg-1) group, E. humifusa extract (18, 54, 162 mg·kg-1·d-1) group and E. humifusa 70% ethanol extract (423 mg·kg-1) group were continuously administered for 14 days. Urine volume was detected 24 h after modeling and administration. Serum creatinine (SCr), Blood urea nitrogen (BUN), 24-hour urine protein (24 hUTP) and uric acid (UA) content; the contents of malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), carbon monoxide synthase (NOS) and lactate dehydrogenase (LDH) in kidney were measured. The levels of interleukin (IL)-6 and tumor necrosis factor (TNF)-α in serum were detected by enzyme linked immunosorbent assay(ELISA) kit. The pathological changes of renal tissue were detected by hematoxylin-eosin (HE) and Masson staining. Western blot was used to detect the expression of PI3K/protein kinase B(Akt)/NF-κB signaling pathway-related proteins. ResultsIn this study, 13 active components such as kaempferol, luteolin, apigenin, gallic acid and quercetin were screened and identified from E. humifusa. Through bioinformatics analysis, these components and AKI have a total of 289 targets, of which 62 are core targets, including Akt1, TNF, tumor protein p53(TP53) and IL-1β. These targets are mainly involved in the regulation of biological processes such as NF-κB signaling pathway, HIF-1 signaling pathway, TNF signaling pathway, PI3K/Akt signaling pathway and mitogen-activated protein kinase(MAPK) signaling pathway. In animal experiments, we successfully constructed a GM-induced AKI model in rats. Compared with the model group, E. humifusa extract could significantly reduce the levels of 24 hUTP, BUN and SCr in rats (P<0.01), indicating its improvement effect on renal function. In addition, the extract of E. humifusa also significantly reduced LDH activity and MDA content in rat kidney tissue (P<0.05, P<0.01), and significantly increased SOD, NOS activity and GSH content (P<0.05), indicating that the extract of E. humifusa has the potential of anti-oxidation and protection of renal function. Further analysis of inflammatory factors showed that the levels of IL-6 and TNF-α in serum of rats treated with E. humifusa extract were significantly decreased (P<0.01), indicating that E. humifusa extract had anti-inflammatory effects. In addition, the extract of E. humifusa can also regulate the protein expression of PI3K/Akt/NF-κB signaling pathway, which further confirmed its mechanism of reducing GM-induced AKI. ConclusionThe extract of E. humifusa has a significant therapeutic effect on acute kidney injury through its multi-component and multi-target mechanism. Its effect is reflected in improving renal function, anti-oxidation, anti-inflammation and regulating immune response. These findings provide a scientific basis for the application of E. humifusa in the treatment of acute kidney injury, and point out the direction for future drug development and clinical research.
5.The impact of donor reentry experience on blood donation return intention
Shangwu LI ; Yao GUAN ; Yuan YUAN ; Jing CHEN ; Minghua TAN ; Jia LUO
Chinese Journal of Blood Transfusion 2026;39(5):636-642
Objective: To explore the impact of donor reentry experience, specifically among those with a single reactive serological result who completed the reentry process, on their willingness to return for future blood donation, and to examine the mediating roles of blood donation knowledge and trait anxiety. Methods: A cross-sectional survey was conducted between November and December 2025. A total of 386 blood donors from the Changsha Blood Center were categorized into a reentry group (n=123) and a control group (n=263). Data on demographic characteristics, blood donation knowledge (BDKQ), trait anxiety (STAI-T), and blood donation return intention (BDRIS) were collected via questionnaires. SPSS 27.0 and AMOS 28.0 were used for statistical analyses, including independent-samples t-test, one-way ANOVA, multiple linear regression, and path analysis for mediating effect testing. Results: There were statistically significant differences in age, occupation, education level, monthly income and donation frequency between the reentry group and the control group (all P<0.05). The reentry group scored significantly higher in blood donation knowledge and blood donation return intention than the control group (both P<0.05). The mean BDRIS score was 11.51±3.62, indicating a relatively high intention to return. Blood donation knowledge was significantly negatively correlated with trait anxiety (r=-0.15, P<0.05) and positively correlated with blood donation return intention (r=0.19, P<0.05); trait anxiety was significantly negatively correlated with blood donation return intention (r=-0.33, P<0.05). Significant differences in BDRIS scores were found based on group (reentry vs control), age, and number of previous donations (all P<0.05). Multiple linear regression showed that BDKQ positively predicted BDRIS (β=0.11, P<0.05), while STAI-T negatively predicted BDRIS (β=-0.27, P<0.05). Path analysis further revealed that the reentry experience had no direct effect on the intention to return. However, it exerted a positive influence through two indirect pathways: 1) a simple mediating effect via increased blood donation knowledge (β=0.17, accounting for 25.0% of the total effect), and 2) a chain mediating effect through "increased blood donation knowledge → decreased trait anxiety" (β=0.05, accounting for 8.1% of the total effect). The model fit indices reached the ideal fitting criteria. Conclusion: The donor reentry experience does not directly enhance the intention to return for blood donation. Rather, it may exert an indirect positive influence by increasing blood donation knowledge and through the sequential pathway of "increased knowledge → decreased trait anxiety". Blood collection institutions should leverage the reentry process as an opportunity for education and psychological support to improve donor retention rate.
6.Analysis of Nitrofuran Metabolites in Meat by Solid-Phase Extraction of Porous Organic Polymer Combined with Liquid Chromatography-Tandem Mass Spectrometry
Yao CHEN ; Ying-Jiao DONG ; Jia-Yi LI ; Rui-Jie WANG ; Zhi-Kai HONG ; Guan-Hua WANG
Chinese Journal of Analytical Chemistry 2025;53(5):804-813
In this work,with tris(4-aminophenyl)amine(TAPA)and 1,3,5-tris(4-formylphenyl)benzene(TFPB)as monomers,an imine-type porous organic polymer,TAPA-TFPB,was synthesized using a simple method under the catalysis of acetic acid.The material TAPA-TFPB was used as solid-phase extraction adsorbent and combined with ultra-performance liquid chromatography/quadrupole time-of-flight-tandem mass spectrometry(UHPLC-QTOF-MS)to establish a detection method for four kinds of nitrofuran metabolites(NFMs)residues in meat samples.The parameters of the adsorbent dosage,the pH value and volume of sample,and the type and volume of washing and eluent solvents were optimized,respectively.Under the optimal extraction conditions,low detection limits(0.11-1.60 μg/kg)were achieved for four kinds of NFMs.At three different spiked levels,the intra-day and inter-day precisions(Relative standard deviations)were 2.8% -10.9% and 4.3% -16.2%,respectively,and the spiked recoveries were 72.0% -107.2%.The results showed that the method chould be used for efficient extraction and analysis of trace NFMs residues in meat samples,indicating that TAPA-TFPB was a kind of promising SPE adsorbent.
7.Effectiveness of telephone and WeChat follow-up in patients discharged from hospital with retained chest drains after stage Ⅲ tuberculous empyema surgery
Ye YUAN ; Ning WANG ; Shuang JIA ; Dan LI ; Jun ZHOU ; Xiaojun YAO
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2025;32(10):1432-1437
Objective To investigate the efficacy of combined telephone and WeChat follow-up for patients discharged with an indwelling closed thoracic drainage tube after surgery for stage Ⅲ tuberculous empyema. Methods Patients with stage Ⅲ tuberculous empyema who were discharged with an indwelling drainage tube from the Department of Thoracic Surgery, Public Health Clinical Center of Chengdu, between November 2021 and November 2022 were enrolled in this study. They were divided into an observation group (combined telephone and WeChat follow-up) and a control group (telephone-only follow-up). The quality of life (QoL), treatment adherence, and recovery outcomes were compared between the two groups. Results A total of 81 patients were included. The observation group consisted of 49 patients (31 males, 18 females) with a mean age of (38.63±15.86) years. The control group consisted of 32 patients (27 males, 5 females) with a mean age of (36.91±17.33) years. The observation group showed significantly better postoperative QoL outcomes in the domains of physical functioning, emotional functioning, physical symptoms, global health status, and overall QoL compared to the control group (all P<0.05). Regarding treatment adherence, the observation group demonstrated superior performance in daily activity duration, use of a respiratory trainer, and adherence to coughing exercises compared to the control group (all P<0.001). The duration of chest tube indwelling was significantly shorter in the observation group (P<0.001). Furthermore, the observation group showed better recovery in albumin and hemoglobin levels (both P<0.05). Conclusion A combined telephone and WeChat follow-up approach can significantly improve the QoL and treatment adherence for patients discharged with an indwelling drainage tube. This method effectively shortens the duration of postoperative tube drainage, promotes nutritional recovery, and accelerates overall postoperative rehabilitation.
8.Clinical Observation of Modified Zhigancao Tang in Treating Patients with Liver and Kidney Deficiency of Parkinson's Disease and Its Effect on Neuronal Signal-related Proteins
Yifo WEI ; Furong LYU ; Jia YAO ; Guonian LI ; Xianyi LUO ; Meng LUO ; Zhengzheng WEN ; Qiuqi LI ; Yihan LIU ; Linlin YANG ; Rui ZUO ; Wenxin DANG ; Fang MI ; Xiaoyan WANG ; Zhigang CHEN ; Fan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):166-173
ObjectiveMicrotube associated protein-2 (MAP-2), alpha-tubulin (α-tubulin), and synaptophysin (SYP) are important proteins in neuronal signal communication. This paper observed the effects of modified Zhigancao Tang on the expression of serum α-Synuclein (α-Syn) and its oligomers, MAP-2, α-tubulin, and SYP of patients with liver and kidney deficiency of Parkinson's disease (PD), analyzed their correlation, and evaluated the therapeutic effect of modified Zhigancao Tang in patients with liver and kidney deficiency of PD based on α-Syn transmission pathway mediated by neuronal communication in vivo. MethodsA total of 60 patients with PD who met the inclusion criteria were randomly divided into a treatment group (30 cases) and a control group (30 cases). Both groups were treated on the basis of PD medicine, and the treatment group was treated with modified Zhigancao Tang. Both groups were treated for 12 weeks. The changes in UPDRS score, TCM syndrome score, and expression of serum α-Syn and its oligomers, MAP-2, α-tubulin, and SYP were observed before and after 12 weeks of treatment in each group. The correlation between the above-mentioned serum biological indexes and the levels of serum α-Syn and its oligomers was analyzed. ResultsAfter treatment, the TCM syndrome score, UPDRS score, UPDRS-Ⅱ score, and UPDRS-Ⅲ score of the treatment group were significantly decreased (P<0.05, P<0.01). The UPDRS score, UPDRS-Ⅱ score, and UPDRS-Ⅲ scores in the treatment group were significantly decreased compared with those in the control group after treatment (P<0.05). After treatment, the total effective rate of the control group was 63.3% (19/30), and that of the treatment group was 86.7% (26/30). The clinical effect of the observation group was better than the control group (Z=-2.03, P<0.05). The total effective rate of the observation group was better than that of the control group, and the difference was statistically significant (χ2=5.136, P<0.05). After treatment, the oligomer level of serum α-Syn and MAP-2 level in the treatment group were significantly decreased (P<0.05, P<0.01). The levels of serum α-Syn and its oligomers, as well as α-tubulin in the treatment group, were significantly decreased compared with those in the control group after treatment (P<0.05, P<0.01). Serum α-Syn was correlated with serum MAP-2 and α-Syn oligomer in patients with PD (P<0.05, P<0.01) but not correlated with serum SYP . Serum α-Syn oligomers of patients with PD were correlated with serum MAP-2 and α-tubulin (P<0.05, P<0.01) but not correlated with serum SYP level. Serum SYP of patients with PD was correlated with serum MAP-2 (P<0.05). ConclusionModified Zhigancao Tang has a therapeutic effect on patients with liver and kidney deficiency of PD by inhibiting the production of α-Syn oligomers and intervening α-Syn microtubule transport pathway in vivo.
9.Interpretation of 2024 ESC guidelines for the management of elevated blood pressure and hypertension
Yu CHENG ; Yiheng ZHOU ; Yao LÜ ; ; Dongze LI ; Lidi LIU ; Peng ZHANG ; Rong YANG ; Yu JIA ; Rui ZENG ; Zhi WAN ; Xiaoyang LIAO
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2025;32(01):31-40
The European Society of Cardiology (ESC) released the "2024 ESC guidelines for the management of elevated blood pressure and hypertension" on August 30, 2024. This guideline updates the 2018 "Guidelines for the management of arterial hypertension." One notable update is the introduction of the concept of "elevated blood pressure" (120-139/70-89 mm Hg). Additionally, a new systolic blood pressure target range of 120-129 mm Hg has been proposed for most patients receiving antihypertensive treatment. The guideline also includes numerous additions or revisions in areas such as non-pharmacological interventions and device-based treatments for hypertension. This article interprets the guideline's recommendations on definition and classification of elevated blood pressure and hypertension, and cardiovascular disease risk assessment, diagnosing hypertension and investigating underlying causes, preventing and treating elevated blood pressure and hypertension. We provide a comparison interpretation with the 2018 "Guidelines for the management of arterial hypertension" and the "2017 ACC/AHA guideline on the prevention, detection, evaluation, and management of high blood pressure in adults."
10.Dehydrodiisoeugenol resists H1N1 virus infection via TFEB/autophagy-lysosome pathway.
Zhe LIU ; Jun-Liang LI ; Yi-Xiang ZHOU ; Xia LIU ; Yan-Li YU ; Zheng LUO ; Yao WANG ; Xin JIA
China Journal of Chinese Materia Medica 2025;50(6):1650-1658
The present study delves into the cellular mechanisms underlying the antiviral effects of dehydrodiisoeugenol(DEH) by focusing on the transcription factor EB(TFEB)/autophagy-lysosome pathway. The cell counting kit-8(CCK-8) was utilized to assess the impact of DEH on the viability of human non-small cell lung cancer cells(A549). The inhibitory effect of DEH on the replication of influenza A virus(H1N1) was determined by real-time quantitative polymerase chain reaction(RT-qPCR). Western blot was employed to evaluate the influence of DEH on the expression level of the H1N1 virus nucleoprotein(NP). The effect of DEH on the fluorescence intensity of NP was examined by the immunofluorescence assay. A mouse model of H1N1 virus infection was established via nasal inhalation to evaluate the therapeutic efficacy of 30 mg·kg~(-1) DEH on H1N1 virus infection. RNA sequencing(RNA-seq) was performed for the transcriptional profiling of mouse embryonic fibroblasts(MEFs) in response to DEH. The fluorescent protein-tagged microtubule-associated protein 1 light chain 3(LC3) was used to assess the autophagy induced by DEH. Western blot was employed to determine the effect of DEH on the autophagy flux of LC3Ⅱ/LC3Ⅰ under viral infection conditions. Lastly, the role of TFEB expression in the inhibition of DEH against H1N1 infection was evaluated in immortalized bone marrow-derived macrophage(iBMDM), both wild-type and TFEB knockout. The results revealed that the half-maximal inhibitory concentration(IC_(50)) of DEH for A549 cells was(87.17±0.247)μmol·L~(-1), and DEH inhibited H1N1 virus replication in a dose-dependent manner in vitro. Compared with the H1N1 virus-infected mouse model, the treatment with DEH significantly improved the body weights and survival time of mice. DEH induced LC3 aggregation, and the absence of TFEB expression in iBMDM markedly limited the ability of DEH to counteract H1N1 virus replication. In conclusion, DEH exerts its inhibitory activity against H1N1 infection by activating the TFEB/autophagy-lysosome pathway.
Influenza A Virus, H1N1 Subtype/genetics*
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Animals
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Autophagy/drug effects*
;
Humans
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Mice
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Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics*
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Influenza, Human/metabolism*
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Lysosomes/metabolism*
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Orthomyxoviridae Infections/genetics*
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Eugenol/pharmacology*
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Antiviral Agents/pharmacology*
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Virus Replication/drug effects*
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A549 Cells
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Male

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