1.Compact Fundus Imaging System Using Shack-Hartmann Wavefront Sensing for High-speed Auto-focus
Zhe-Kai LIN ; Long CHEN ; Geng-Yong ZHENG ; Jin-Tian HUANG ; Jia-Xin DONG ; Shang-Pan YANG ; Wen-Zheng DING ; Ding-An HAN ; Xue-Hua WANG ; Ya-Guang ZENG
Progress in Biochemistry and Biophysics 2026;53(4):1076-1086
ObjectiveThe widespread adoption of portable fundus cameras for primary care and community screening is hindered by limitations in current autofocus(AF) technologies. Image-based methods relying on sharpness evaluation require iterative searches, resulting in slow convergence, while projection-based techniques are susceptible to optical artifacts and calibration errors. To address these challenges, this study introduces a novel AF system based on direct wavefront sensing, designed to deliver simultaneous high speed, high precision, and operational robustness within the compact form factor essential for portable ophthalmic devices. MethodsOur approach fundamentally reimagines the AF process by directly measuring the ocular wavefront aberration. We developed a custom portable fundus camera integrating a miniaturized Shack-Hartmann wavefront sensor (SHWS) into the optical path. An 850 nm laser diode projects a point source onto the retina via oblique illumination to minimize corneal reflections. Light scattered from this spot carries the eye’s refractive error through the imaging optics and is directed to the SHWS, positioned at a plane optically conjugate to the primary color CMOS imaging sensor. A microlens array within the SHWS samples the incident wavefront, generating a pattern of focal spots on a CCD. Real-time centroid analysis of these spots provides a map of local wavefront slopes. These measurements are processed through a singular value decomposition (SVD) algorithm to fit a Zernike polynomial basis set, enabling real-time reconstruction of the wavefront phase. The defocus component (S) is extracted from the second-order Zernike coefficients, providing a direct, quantitative measure of the refractive error in diopters. This value serves as a precise error signal in a closed-loop control system, which commands a voice-coil actuated focusing lens to its null position in a single, deterministic step, eliminating the need for iterative search algorithms. ResultsComprehensive evaluation demonstrated the system’s high performance. Testing on a calibrated model eye (OEMI-7) established a highly linear relationship between the computed defocus S and the focusing lens position across a ±20 Diopter (D) compensation range, achievable within a 5 mm mechanical travel. The system achieved a focusing precision of 0.08 D, corresponding to an 18-fold improvement over a conventional projection spot-size method tested under identical conditions. The total focus acquisition time, encompassing wavefront measurement, computation, and lens actuation, averaged under 0.5 s. Clinical validation with 25 human volunteers (50 eyes, refractive range -15 D to +10 D) confirmed practical efficacy. The wavefront-sensing AF succeeded in 92% of attempts with a mean time of 0.5 s, substantially outperforming a projection-based benchmark which achieved only a 32% success rate with an average time of 4.25 s. The system provided instantaneous directional guidance and maintained stability during minor ocular movements. Objective assessment of image quality, via amplitude contrast of retinal vasculature, showed consistent and significant enhancement following AF correction across the entire tested diopter range. ConclusionThis work successfully implements and validates a direct wavefront-sensing autofocus paradigm for portable fundus cameras. By directly quantifying and compensating for the optical defocus aberration, this method bypasses the fundamental limitations of image-processing and projection-based techniques, enabling rapid, precise, and deterministic diopter compensation. The developed system delivers an exceptional combination of a wide operational range (±20 D), high accuracy (0.08 D), fast convergence (0.5 s), and a compact physical footprint. This technology provides a practical and high-performance focusing solution capable of enhancing the reliability, throughput, and diagnostic utility of portable retinal imaging in large-scale screening applications. Future efforts will be directed towards system cost optimization and performance adaptation for diverse ocular conditions.
2.Exploring Mechanisms of Erchentang in Repairing Ileal Immune Barrier and Reducing Weights of Diet-induced Obese Mice Based on Single-cell Transcriptomics
Jiawei CHEN ; Maohui LIU ; Jilan CHEN ; Jiushuang ZHU ; Yingxiu MEI ; Yue JIN ; Xiuwen XIA ; Weijun DING
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):226-236
ObjectiveTo investigate the effects of Erchentang (ECD) on the body weight of the mouse model of simple obesity induced by a high-fat diet (HFD) and decipher the underlying mechanisms. MethodsFirstly, single-cell transcriptomics (Sc-RNAseq) was employed to analyze the transcriptional changes in the ileum tissue of mice in the normal group and model group. Then, a mouse model of simple obesity was established with a high-fat diet. The successfully modeled mice were randomly allocated into the following four groups (n=8): model, low-dose (7.5 g·kg-1) ECD, medium-dose (15 g·kg-1) ECD, and high-dose (30 g·kg-1) ECD. Additionally, 8 mice of the same age were selected as the normal group. The body weight was measured at fixed time points during the 4-week gavage period. The overall efficacy of ECD in alleviating obesity was evaluated through glucose tolerance testing, behavioral analysis, hematoxylin-eosin (HE) staining, and biochemical testing. Protein docking was employed to predict the degree of binding between corresponding proteins. Molecular docking was employed to predict the binding degree between key components of ECD and target proteins. Real-time PCR was employed to determine the mRNA levels of tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), interleukin-1β (IL-1β), CD68, CD206, zonula occludens-1 (ZO-1), and Claudin-5 in the ileum. Immunofluorescence staining was used to observe the expression and distribution of Claudin-5 and ZO-1. ResultsThe Sc-RNAseq results indicated that the differentially expressed genes of immune cells in the model group in comparison with the normal group were primarily enriched in biological functions related to lipid metabolism and inflammatory metabolism. Additionally, these genes were associated with the janus kinases(JAK)/signal transducers and activators of transcription (STAT) signaling pathway, an inflammation-related pathway. Compared with the normal group, the model group showed increases in body weight (P<0.01) and blood glucose level (P<0.01), a decrease in limb strength (P<0.01), an increase in liver weight (P<0.05), and elevated serum alanine amino-transferase (ALT) and aspartate transferase (AST) levels (P<0.05, P<0.01). Additionally, the model group exhibited increased hepatic fat vacuoles, notably enlarged adipocytes in the epididymal and inguinal white adipose tissue, and increased inflammation. Compared with the model group, ECD groups showed reduced body weights (P<0.01) and blood glucose levels (P<0.01), increased limb strength (P<0.05, P<0.01), decreased liver weights (P<0.05, P<0.01), and declined serum ALT and AST levels (P<0.05, P<0.01). Additionally, ECD reduced hepatic fat vacuoles and the adipocyte volume in the epididymal and inguinal white adipose tissue, and alleviated inflammation. Potential interactions existed between CD68 and ZO-1/Claudin-5, as well as between CD206 and ZO-1/Claudin-5. The key components of ECD, nobiletin, diosmetin, and naringenin, all demonstrated strong binding affinity with the target proteins ZO-1 and Claudin-5. Compared with the normal group, the model group exhibited up-regulated mRNA levels of the pro-inflammatory cytokines TNF-α, iNOS, IL-1β, and CD68 (P<0.05, P<0.01) and down-regulated mRNA levels of the anti-inflammatory cytokine CD206 (P<0.01) and the tight junction proteins Claudin-5 and ZO-1 (P<0.05, P<0.01). In comparison with the model group, the ECD groups showed down-regulated mRNA levels of TNF-α, iNOS, IL-1β, and CD68 (P<0.05, P<0.01) and up-regulated mRNA levels of CD206, Claudin-5, and ZO-1 (P<0.05, P<0.01). Compared with the normal group, the model group exhibited down-regulated expression of tight junction proteins Claudin-5 and ZO-1 (P<0.01). Compared with the model group, ECD groups showed up-regulated expression of Claudin-5 and ZO-1 (P<0.05, P<0.01). ConclusionECD can significantly ameliorate HFD-induced obesity and excessive body weight gain in mice by improving the inflammatory microenvironment in the ileum and further restoring the integrity of the impaired ileal barrier.
3.Differences in deltamethrin resistance and kdr gene mutation in Culex tritaeniorhynchus population in and outside the Yellow Sea wetland
Xiao-er ZHANG ; Zhi-ming WU ; Ye TIAN ; Qian CUI ; Yu-qian JI ; Huan WANG ; Shu-juan YANG ; Yi-chao ZHAO ; Yu WANG ; Hua-yu YIN ; Yu DING ; Guo-jin YAN ; Min-sen ZHAO ; Shou-gang ZHANG ; Bing-dong SONG ; Hong-na CHEN ; Jian GAO ; Wei-fang YANG ; Yu-fu ZHANG ; Hui LIU ; Hong-liang CHU
Acta Parasitologica et Medica Entomologica Sinica 2026;33(2):101-107
Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.
4.Evaluation of snail control effect in Anqing small environment renovation project
Qiong CHU ; Ping DING ; Jin-fu YAO ; Xing WANG
Acta Parasitologica et Medica Entomologica Sinica 2026;33(1):9-18
Objective The aim was to evaluate the effects of efforts directed toward controlling Oncomelania hupensis snails on small environmental modification projects in Anqing City to provide a reference for comprehensively controlling O. hupensis in hilly areas where schistosomiasis is endemic. Methods Field and retrospective surveys were conducted to collect data using questionnaires. The data from 35 questionnaires on small-scale environmental modification projects in Anqing City and the effects of snail obtained from monitoring forms were sorted. The data were descriptively analyzed using basic information on environmental modifications and investment costs. The Wilcoxon signed-rank test was used to compare snail status before and after environmental interventions. The Friedman test was applied to evaluate the changes in snail populations in the years after environmental intervention. Non-parametric independent sample tests were adopted to compare differences in the effects of different environmental interventions, environmental characteristics, and between project and control areas. Results A total of 35 environments altered from 2017 to 2023, including the creation of 29 ditches,1 dry land, and 5 pond-weirs. The main environmental interventions included hardening of ditches and pond-weirs as well as tilling with intercropping. The average investment costs of mortared rubble masonry hardening and tilling with intercropping were the highest(147.10 yuan/m2)and lowest(4.62 yuan/m2), respectively. The pre-intervention snail-infested area was 78 900 m2, which decreased to 18 100 m2, a decrease of 77.06%. After environmental intervention, the density and rate of living snails significantly decreased on frames with snails(P<0.001). In ditch environments, the smoothness of the ditch walls and the presence of snails in the upper reaches did not significantly decrease the living snail density; however, the presence of cracks on the inner side of the ditches significantly impacted the decline in the rate(P<0.001). No significant difference was found in the rates at which the living snail density or snail frame occurrence decreased among different environmental modification method. The effects of brick masonry, concrete, and mortared rubble masonry on snail control were significantly stronger than those of chemical snail control(P<0.01), whereas no significant difference was found in the decline in these two rates between project areas using other modification method and control areas using chemical snail control. Conclusions Schistosomiasis-prevention-oriented environmental interventions effectively eliminate snails from hilly snail-infested areas. Cost-effective modification method should be selected based on the environmental type, and post-modification monitoring and maintenance must be strengthened to further increase snail control effectiveness.
5.Objective Indicators and Evaluation System for Excellent Shape and High Quality of Gardeniae Fructus
Xiulin YUAN ; Shiwen KANG ; Zhihao LIU ; Ding HUANG ; Sen YE ; Fan LI ; Bocheng WANG ; Yan JIN ; Yuan YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(18):27-35
ObjectiveTo establish an objective evaluation system based on the quality characteristics of Gardeniae Fructus without relying on the origin label according to the theory of excellent shape, high quality, and superior effect of authentic medicinal materials, thus providing a scientific basis for the standardization of Gardeniae Fructus. MethodsDifferent batches of Gardeniae Fructus samples were collected from different producing areas. Through herbal textual research and a review of modern literature, fruit length, maximum fruit diameter, and chroma were selected as quantitative indicators to characterize the traditional appearance traits of Gardeniae Fructus. The above indicators were measured by a vernier caliper and a spectrophotometer. Through classical famous prescription data and literature analysis, total iridoid glycosides, total crocus glycosides, and total flavonoids were selected as the core component indicators related to efficacy, and their content was determined. Finally, through cluster analysis, the threshold ranges of the quantitative indicators of excellent shape and high quality were determined, and the applicability of the established evaluation system was verified by the determination of the indicators for each batch of medicinal materials. ResultsThe excellent shape indicators of Gardeniae Fructus were the fruit length-to-maximum fruit diameter ratio ≤1.86, 0°≤h≤45°, and L*≥ 37.32. The content of total iridoid glycosides, total crocus glycosides, and total flavonoids should be no less than 2.2%, 0.44%, and 0.028%, respectively. The measurement results of different batches of Gardeniae Fructus showed that the evaluation system could effectively distinguish the Gardeniae Fructus samples in line with the traditional cognition of excellent shape and high quality. ConclusionThis study preliminarily constructs an excellent shape and high quality-based quality evaluation system for Gardeniae Fructus, which links appearance traits with chemical indicators and provides a methodological basis for the quality control and scientific grading of this herbal medicine.
6.HydroMg alleviates doxorubicin-induced chronic cardiotoxicity by attenuating oxidative stress and improving mitochondrial function
Lu ZHANG ; Xiurui MA ; Yawei JIN ; Yuning ZHANG ; Xiong GAO ; Mohan LI ; Ze YUAN ; Yihua LU ; Wenjiang DING ; Zhiguang DING ; Xiaolei SUN ; Chunxiao ZHANG ; Jian AN
Chinese Journal of Clinical Medicine 2026;33(4):582-591
Objective To investigate the protective effect and underlying mechanisms of HydroMg, a novel sustained-release hydrogen donor, in a model of doxorubicin (DOX)-induced chronic cardiotoxicity. Methods H9C2 cells were treated with 1 μmol/L DOX and/or 1 μg/mL HydroMg, and mitochondrial membrane potential, reactive oxygen species (ROS) levels, and apoptosis were measured. In the animal experiment, mice were divided into control, HydroMg, DOX, and DOX+HydroMg groups. The HydroMg group received intraperitoneal injection of HydroMg (100 mg/kg). Chronic cardiotoxicity was established by intraperitoneal injection of DOX (5 mg/kg every week) for four consecutive weeks; In the DOX+HydroMg group, HydroMg (100 mg/kg) was administered intraperitoneally 4–5 h prior to each DOX injection. Cardiac function and remodeling were evaluated by echocardiography and heart weight index (HWI). The long-term survival of mice were analyzed. Subsequently, transcriptome sequencing was performed on myocardial tissues from the DOX group to identify key molecular alterations compared with controls. Results At the cellular level, HydroMg treatment effectively reversed DOX-induced mitochondrial dysfunction and oxidative stress, as evidenced by restored membrane potential, reductions in total ROS and mitochondrial superoxide levels by approximately 30% and 9%, respectively, and a decrease in the apoptosis rate from 34.13% to 18.27%. In the animal model, compared with the DOX group, HydroMg intervention increased left ventricular ejection fraction (LVEF) and fractional shortening (LVFS) by 9.62% and 5.82%, respectively, attenuated the reduction in HWI (4.41 mg/mm vs 3.53 mg/mm), and improved the 4-week survival rate (80% vs 60%). Transcriptomic analysis revealed that the core molecular features of DOX-induced cardiotoxicity, including widespread suppression of oxidative phosphorylation and antioxidant pathways. Conclusions HydroMg protects against DOX-induced chronic cardiotoxicity by alleviating oxidative stress and improving mitochondrial function, and provides a potential therapeutic strategy against DOX-related cardiotoxicity.
7.MADS-Net: a Multi-scale Attention and Dynamic Sparse Mask Fusion Network for Electrical Impedance Tomography Image Reconstruction in Breast Screening
Cheng-Tao YU ; Jia-Hui WANG ; Song-Pei HU ; Huai-Jin CHEN ; Li DING ; Xia YE ; Jia-Feng YAO
Progress in Biochemistry and Biophysics 2026;53(9):2480-2496
ObjectiveTo address the key challenges in breast electrical impedance tomography (EIT) reconstruction including the severe ill‑posedness of the inverse problem, limited measurement information from the electrodes, low contrast of small lesions, as well as the common drawbacks of traditional reconstruction methods such as boundary blurring, low spatial resolution, and weak small-target detection capability, and to overcome the limitations of existing deep learning approaches namely low training efficiency, insufficient physical consistency, and substantial information loss during the low‑to‑high dimensional mapping process, this study proposes a multi‑scale attention and dynamic sparse mask fusion network (MADS‑Net). The proposed method aims to enhance high‑resolution reconstruction accuracy, small-lesion detectability, noise robustness, and computational efficiency in breast‑screening-oriented EIT, thereby providing a feasible solution for the application of EIT in breast functional imaging. MethodsThe proposed network adopts a strategy of “locate first, reconstruct later”. It realizes adaptive allocation of computational resources through a dynamic mask module, alleviates information loss in the low-to-high dimensional mapping process via a multi-scale attention mechanism, and embeds physical constraints to improve the physical rationality of reconstruction results. The performance of the proposed method was validated through systematic numerical simulations, agar phantom experiments, and porcine tissue experiments. Comparisons were made with mainstream methods including the Gauss-Newton algorithm, convolutional neural networks, U-Net, and Deep D-Bar. The image correlation coefficient (ICC), root mean square error (RMSE), and peak signal-to-noise ratio (PSNR) were adopted as quantitative evaluation metrics. Furthermore, lesion‑level evaluation metrics were introduced to assess the capability of different methods in reconstructing abnormal target boundaries and small‑sized target regions. ResultsExperimental results demonstrate that MADS-Net outperforms all comparison methods across all test scenarios. In numerical simulations, it achieves higher accuracy in target morphology and spatial location recovery as well as better background suppression. In agar phantom experiments, its detection and imaging reliability for small targets is significantly superior to the comparison methods. In porcine tissue experiments, the model maintains good generalization performance and can adapt to complex biological tissue imaging scenarios. Quantitative results show that MADS-Net achieves an average ICC of 0.917 6±0.048 8, an RMSE as low as0.060 6±0.031 6, and a PSNR of (23.893 9±2.685 3) dB. It successfully detects tiny targets as small as 5 mm within a 100 mm diameter sensing field, reduces the total training duration by 23.5% compared with the baseline method, and maintains stable reconstruction performance even under strong noise interference. ConclusionThe results demonstrate that MADS-Net achieves favorable EIT reconstruction performance in numerical simulations, agar phantom experiments, and porcine tissue experiments, even under common practical imaging challenges including low signal-to-noise ratio and blurred tissue boundaries. This study provides a methodological reference for further research on EIT in breast-screening-related scenarios and other medical functional imaging tasks, and also offers a low-cost, radiation-free alternative for breast lesion screening that aligns with the clinical demand for non-invasive diagnostic tools. It should be noted that real clinical breast data have not yet been used for validation in this study, and the clinical applicability of the proposed method requires further investigation. Future work will prioritize collecting multi-center real clinical breast datasets, and further optimize the model’s generalization ability to adapt to individual differences in breast tissue composition to promote its clinical application.
8.Mechanism of Paeoniflorin Regulating TLR4/NF-κB-IRS1 Inflammation-insulin Signaling Axis to Improve Metabolic Dysfunction Associated Fatty Liver Disease Combined with Insulin Resistance
Luyu LI ; Yiming FAN ; Wenlong YU ; Yiteng ZHANG ; Yaorui HU ; Huanxin DING ; Chuxuan LIU ; Xin JIN ; Hongyu ZHANG ; Qian XU ; Guangyong ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(21):146-156
ObjectiveTo investigate the therapeutic effect of paeoniflorin on metabolic dysfunction-associated fatty liver disease (MASLD) combined with insulin resistance (IR), and to elucidate its regulatory mechanisms on the Toll-like receptor 4 (TLR4)/nuclear transcription factor (NF)-κB-insulin receptor substrate 1 (IRS1) inflammation-insulin signaling axis. MethodsMASLD mouse model was established using a 60% high-fat diet. Mice were randomly assigned to a normal control group, a MASLD model group, and paeoniflorin groups (low dose: 25 mg·kg-1·d-1, medium dose: 50 mg·kg-1·d-1, high dose: 100 mg·kg-1·d-1), receiving intragastric administration for consecutive 12 weeks. Concurrently, a palmitic acid/oleic acid (PA/OA)-induced lipid deposition model was established in HepG2 cells. Cell counting kit-8 (CCK-8) assay was performed to determine the optimal concentrations of PA/OA and paeoniflorin for intervention. Mice body weight, liver index, and serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglycerides (TG), and total cholesterol (TC) were measured. Fasting blood glucose (FBG) and fasting insulin (FINS) levels were measured, and the homeostatic model assessment of insulin resistance (HOMA-IR) was calculated, while an oral glucose tolerance test (OGTT) was performed to evaluate glucose metabolism. Hematoxylin and eosin (HE) staining was used to observe pathological changes and inflammatory cell infiltration in liver tissue, and oil red O staining was used to evaluate lipid droplet deposition in liver tissue and HepG2 cells. Potential targets were screened using network pharmacology and molecular docking. Western blot analysis was performed to detect the expression of HSP90AA1, TLR4, p-NF-κB p65, p-IRS1, and p-Akt proteins in liver tissue and HepG2 cells. ResultsCompared with the normal control group, mice in the MASLD model group exhibited significantly increased body weight, liver index, and serum levels of ALT, AST, TG, and TC, as well as markedly elevated FBG, FINS, and HOMA-IR, and impaired glucose tolerance (P<0.01). HE staining revealed marked hepatic steatosis and inflammatory cell infiltration, while oil red O staining showed a significant increase in lipid droplet deposition. Compared with the MASLD model group, after intervention with various doses of paeoniflorin, the increase in body weight and liver index was reduced, serum ALT, AST, TG, and TC levels decreased, and FBG, FINS, and HOMA-IR were significantly lowered, with glucose tolerance markedly improved (P<0.05, P<0.01), showing a certain dose-dependent trend. HE and oil red O staining results showed a marked reduction in hepatic steatosis and lipid deposition. In vitro experiments demonstrated that, compared with the control group, PA/OA treatment significantly induced increased lipid deposition in HepG2 cells. Compared with the model group, paeoniflorin intervention significantly reduced intracellular lipid droplets, and lipid deposition showed a dose-dependent decreasing trend. Mechanism studies indicated that, compared with the normal control group, the MASLD model group exhibited significantly elevated expression of TLR4, HSP90AA1, and p-NF-κB p65, while p-IRS1 (Ser307) expression was elevated and p-Akt (Ser473) expression was reduced (P<0.01). Compared with the model group, the expression of the aforementioned proteins was significantly reversed in all treatment groups following intervention (P<0.05, P<0.01). ConclusionPaeoniflorin significantly alleviates lipid deposition and insulin resistance in MASLD mice. Its mechanism of action may involve targeting HSP90AA1, TLR4, and NF-κB1 to regulate the TLR4/NF-κB-IRS1 inflammation-insulin signaling axis. This regulation suppresses inflammatory responses and restores insulin signaling, thereby ameliorating abnormalities in glucose and lipid metabolism.
9.Mechanism of Paeoniflorin Regulating TLR4/NF-κB-IRS1 Inflammation-insulin Signaling Axis to Improve Metabolic Dysfunction Associated Fatty Liver Disease Combined with Insulin Resistance
Luyu LI ; Yiming FAN ; Wenlong YU ; Yiteng ZHANG ; Yaorui HU ; Huanxin DING ; Chuxuan LIU ; Xin JIN ; Hongyu ZHANG ; Qian XU ; Guangyong ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(21):146-156
ObjectiveTo investigate the therapeutic effect of paeoniflorin on metabolic dysfunction-associated fatty liver disease (MASLD) combined with insulin resistance (IR), and to elucidate its regulatory mechanisms on the Toll-like receptor 4 (TLR4)/nuclear transcription factor (NF)-κB-insulin receptor substrate 1 (IRS1) inflammation-insulin signaling axis. MethodsMASLD mouse model was established using a 60% high-fat diet. Mice were randomly assigned to a normal control group, a MASLD model group, and paeoniflorin groups (low dose: 25 mg·kg-1·d-1, medium dose: 50 mg·kg-1·d-1, high dose: 100 mg·kg-1·d-1), receiving intragastric administration for consecutive 12 weeks. Concurrently, a palmitic acid/oleic acid (PA/OA)-induced lipid deposition model was established in HepG2 cells. Cell counting kit-8 (CCK-8) assay was performed to determine the optimal concentrations of PA/OA and paeoniflorin for intervention. Mice body weight, liver index, and serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglycerides (TG), and total cholesterol (TC) were measured. Fasting blood glucose (FBG) and fasting insulin (FINS) levels were measured, and the homeostatic model assessment of insulin resistance (HOMA-IR) was calculated, while an oral glucose tolerance test (OGTT) was performed to evaluate glucose metabolism. Hematoxylin and eosin (HE) staining was used to observe pathological changes and inflammatory cell infiltration in liver tissue, and oil red O staining was used to evaluate lipid droplet deposition in liver tissue and HepG2 cells. Potential targets were screened using network pharmacology and molecular docking. Western blot analysis was performed to detect the expression of HSP90AA1, TLR4, p-NF-κB p65, p-IRS1, and p-Akt proteins in liver tissue and HepG2 cells. ResultsCompared with the normal control group, mice in the MASLD model group exhibited significantly increased body weight, liver index, and serum levels of ALT, AST, TG, and TC, as well as markedly elevated FBG, FINS, and HOMA-IR, and impaired glucose tolerance (P<0.01). HE staining revealed marked hepatic steatosis and inflammatory cell infiltration, while oil red O staining showed a significant increase in lipid droplet deposition. Compared with the MASLD model group, after intervention with various doses of paeoniflorin, the increase in body weight and liver index was reduced, serum ALT, AST, TG, and TC levels decreased, and FBG, FINS, and HOMA-IR were significantly lowered, with glucose tolerance markedly improved (P<0.05, P<0.01), showing a certain dose-dependent trend. HE and oil red O staining results showed a marked reduction in hepatic steatosis and lipid deposition. In vitro experiments demonstrated that, compared with the control group, PA/OA treatment significantly induced increased lipid deposition in HepG2 cells. Compared with the model group, paeoniflorin intervention significantly reduced intracellular lipid droplets, and lipid deposition showed a dose-dependent decreasing trend. Mechanism studies indicated that, compared with the normal control group, the MASLD model group exhibited significantly elevated expression of TLR4, HSP90AA1, and p-NF-κB p65, while p-IRS1 (Ser307) expression was elevated and p-Akt (Ser473) expression was reduced (P<0.01). Compared with the model group, the expression of the aforementioned proteins was significantly reversed in all treatment groups following intervention (P<0.05, P<0.01). ConclusionPaeoniflorin significantly alleviates lipid deposition and insulin resistance in MASLD mice. Its mechanism of action may involve targeting HSP90AA1, TLR4, and NF-κB1 to regulate the TLR4/NF-κB-IRS1 inflammation-insulin signaling axis. This regulation suppresses inflammatory responses and restores insulin signaling, thereby ameliorating abnormalities in glucose and lipid metabolism.
10.Molecular Mechanisms of LIGHT Regulating Tumor Vascular Microenvironment and Its Precision Delivery Strategies for Solid Tumor Immunotherapy
Jun-Yi YUAN ; Shao-Xia WANG ; Li-Ming JIN ; Bin-Chun LI ; Guo-Bin DING
Progress in Biochemistry and Biophysics 2026;53(10):2583-2595
The drug resistance in the treatment of solid tumors is closely related to abnormalities in the tumor vascular microenvironment (TVM). The TVM is dynamically composed of multiple stromal, vascular, and immune cell populations together with extracellular matrix. Its structural disorder and functional imbalance can lead to abnormal angiogenesis, perfusion disorders, limited drug delivery, and insufficient immune cell infiltration, thereby creating a microenvironment conducive to tumor progression and treatment resistance. Therefore, the coordinated regulation of TVM through vascular normalization and immune remodeling has become an important research direction for improving the treatment effect of solid tumors. Tumor necrosis factor superfamily member 14 (TNFSF14), also known as LIGHT (homologous to lymphotoxin, inducible, competes with herpes simplex virus (HSV) glycoprotein D for herpesvirus entry mediator (HVEM), expressed by T lymphocyte), is an important molecule connecting tumor vascular remodeling and immune regulation. LIGHT can bind to the lymphotoxin β receptor (LTβR) to activate the NF-κB (nuclear factor kappa B)-related signaling pathway, regulate the functions of vascular endothelial cells and stromal cells, and induce the expression of adhesion molecules and chemokines such as VCAM-1(vascular cell adhesion molecule 1), ICAM-1(intercellular adhesion molecule 1), CCL19 (C-C motif chemokine ligand 19), CCL21 (C-C motif chemokine ligand 21), and CXCL13 (C-X-C motif chemokine ligand 13), promoting the formation of high endothelial venules (HEVs). HEVs can improve the vascular access of immune cells into tumor tissues and further promote the formation of tertiary lymphoid structures (TLS), providing a structural basis for the recruitment of effector T cells and sustained anti-tumor immune responses. At the same time, LIGHT interacts with HVEM to provide costimulatory signals to T cells, enhancing their activation, proliferation, and cytotoxic function. Thus, LIGHT can link tumor vascular normalization, immune cell recruitment, and T cell activation through the dual actions mediated by LTβR and HVEM, forming a coordinated regulation model of “vascular remodeling-immune infiltration-immune activation”, providing new molecular targets for the treatment of solid tumors. Despite LIGHT’s good anti-tumor potential, its therapeutic application is still limited by factors such as insufficient stability in vivo, short half-life, and potential off-target effects due to systemic exposure. In recent years, researchers have developed various strategies for the precise delivery of LIGHT, including fusion proteins, bacterial vectors, viral vectors, nanoparticles, chimeric antigen receptor T cells (CAR-T), and mesenchymal stem cells (MSCs), to enhance the enrichment, expression, or sustained release of LIGHT in tumor sites through different approaches, thereby improving its therapeutic efficacy and safety. This article systematically reviews the structural characteristics, biological functions, and molecular mechanisms mediating TVM remodeling by LIGHT, with a focus on the signal transduction mediated by LTβR and HVEM and their roles in tumor immune regulation. Subsequently, it systematically outlines the molecular mechanisms by which LIGHT regulates the tumor vascular microenvironment and its intrinsic connection with immune cell recruitment and activation, from the perspectives of TVM abnormalities, vascular normalization, HEV formation, and TLS induction. Furthermore, it categorizes and summarizes precise delivery strategies such as LIGHT fusion proteins, bacterial vectors, viral vectors, nanoparticles, CAR-T cells, and MSCs according to delivery vectors and modes of action, and compares their functional characteristics, antitumor effects, and potential advantages and limitations. On this basis, the article further discusses the key issues that LIGHT therapeutic strategies still face in terms of targeting, controllability,in vivo stability, safety, and clinical translation, and anticipates the potential value of its combination with immune checkpoint inhibitors, anti-angiogenic therapy, and other tumor treatment methods. This article aims to deepen the understanding of the antitumor mechanism of LIGHT from the perspective of synergistic regulation of tumor vascular microenvironment remodeling and immune activation, providing theoretical basis and new research ideas for the design of solid tumor treatment strategies based on precise regulation of TVM and the clinical translation of LIGHT-related treatment systems.


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