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.Mitogen-activated protein kinase signaling pathway regulates the development of osteoarthritis:guiding targeted therapy with traditional Chinese medicine
Hao LI ; Hongcheng TAO ; Ping ZENG ; Jinfu LIU ; Qiang DING ; Chicheng NIU ; Kai HUANG ; Hongyu KANG
Chinese Journal of Tissue Engineering Research 2026;30(6):1476-1485
BACKGROUND:Osteoarthritis is pathologically characterized by progressive degeneration of the articular cartilage and abnormal deformation of the subchondral bone.In recent years,with the deepening of medical research,it has been found that the mitogen-activated protein kinases(MAPK)signaling pathway has a regulatory role in inflammatory cell infiltration,inflammatory factor release,and chondrocyte proliferation,which is particularly important for the treatment of osteoarthritis.OBJECTIVE:To briefly review the main research progress in the mechanism of MAPK signaling pathway regulating osteoarthritis in recent years,aiming to provide new ideas for the treatment of osteoarthritis.METHODS:CNKI,WanFang and PubMed databases were searched for relevant literature using the search terms of"mitogen-activated protein kinases,osteoarthritis,extracellular signal-regulated MAP kinases,p38 mitogen-activated protein kinases,JNK mitogen-activated protein kinase"in Chinese and English.Relevant literature published from January 2019 to November 2024 was searched,and 108 articles were finally included for summary analysis.RESULTS AND CONCLUSION:(1)Various stimuli inside and outside the cells activate the MAPK signaling pathway,regulate gene transcription and protein synthesis,and promote the release of inflammatory factors,such as tumor necrosis factor-α,interleukin-1β,and interleukin-6.The release of these inflammatory factors aggravates the progression of osteoarthritis.(2)The active ingredients of traditional Chinese medicine,mainly saponins and flavonoids,as well as Chinese herbal formulas and preparations with the main effects of activating blood circulation and removing blood stasis,tonifying the liver and kidney,can play a therapeutic role in osteoarthritis by inhibiting the MAPK signaling pathway,regulating the release of matrix metalloproteinases,balancing the homeostatic state of osteogenesis and osteoblastogenesis,attenuating the synovial inflammation,decreasing the release of inflammatory factors and inflammatory vesicles,decreasing cellular pyroptosis,promoting autophagy,and ameliorating oxidative stress.(3)Although traditional Chinese medicine has become popular in the treatment of osteoarthritis by virtue of its own advantages of multi-components,multi-targets,multi-pathways,and low side effects,the use of MAPK signaling pathway to guide the treatment of individual osteoarthritis is the difficulty of the technology,which needs to be continuously researched and explored.(4)Therefore,further development of relevant herbal inhibitors that can modulate the MAPK signaling pathway may be a potential drug strategy for the treatment of osteoarthritis in the future.
3.Mitogen-activated protein kinase signaling pathway regulates the development of osteoarthritis:guiding targeted therapy with traditional Chinese medicine
Hao LI ; Hongcheng TAO ; Ping ZENG ; Jinfu LIU ; Qiang DING ; Chicheng NIU ; Kai HUANG ; Hongyu KANG
Chinese Journal of Tissue Engineering Research 2026;30(6):1476-1485
BACKGROUND:Osteoarthritis is pathologically characterized by progressive degeneration of the articular cartilage and abnormal deformation of the subchondral bone.In recent years,with the deepening of medical research,it has been found that the mitogen-activated protein kinases(MAPK)signaling pathway has a regulatory role in inflammatory cell infiltration,inflammatory factor release,and chondrocyte proliferation,which is particularly important for the treatment of osteoarthritis.OBJECTIVE:To briefly review the main research progress in the mechanism of MAPK signaling pathway regulating osteoarthritis in recent years,aiming to provide new ideas for the treatment of osteoarthritis.METHODS:CNKI,WanFang and PubMed databases were searched for relevant literature using the search terms of"mitogen-activated protein kinases,osteoarthritis,extracellular signal-regulated MAP kinases,p38 mitogen-activated protein kinases,JNK mitogen-activated protein kinase"in Chinese and English.Relevant literature published from January 2019 to November 2024 was searched,and 108 articles were finally included for summary analysis.RESULTS AND CONCLUSION:(1)Various stimuli inside and outside the cells activate the MAPK signaling pathway,regulate gene transcription and protein synthesis,and promote the release of inflammatory factors,such as tumor necrosis factor-α,interleukin-1β,and interleukin-6.The release of these inflammatory factors aggravates the progression of osteoarthritis.(2)The active ingredients of traditional Chinese medicine,mainly saponins and flavonoids,as well as Chinese herbal formulas and preparations with the main effects of activating blood circulation and removing blood stasis,tonifying the liver and kidney,can play a therapeutic role in osteoarthritis by inhibiting the MAPK signaling pathway,regulating the release of matrix metalloproteinases,balancing the homeostatic state of osteogenesis and osteoblastogenesis,attenuating the synovial inflammation,decreasing the release of inflammatory factors and inflammatory vesicles,decreasing cellular pyroptosis,promoting autophagy,and ameliorating oxidative stress.(3)Although traditional Chinese medicine has become popular in the treatment of osteoarthritis by virtue of its own advantages of multi-components,multi-targets,multi-pathways,and low side effects,the use of MAPK signaling pathway to guide the treatment of individual osteoarthritis is the difficulty of the technology,which needs to be continuously researched and explored.(4)Therefore,further development of relevant herbal inhibitors that can modulate the MAPK signaling pathway may be a potential drug strategy for the treatment of osteoarthritis in the future.
4.Value of three-dimensional inversion-recovery with real reconstruction sequence using an ultralong repetition time for endolymphatic hydrops
Menglong ZHAO ; Huaili JIANG ; Shujie ZHANG ; Zhuang LIU ; Kai LIU ; Di WU ; Xinsheng HUANG ; Mengsu ZENG
Chinese Journal of Clinical Medicine 2025;32(2):200-206
Objective To evaluate the value of an optimized three-dimensional inversion-recovery with real reconstruction (3D-real IR) sequence with a longer repetition time (TR, 16 000 ms) based on modulated flip angle technique in refocused imaging with extended echo train (MATRIX) in the endolymphatic hydrops (EH) imaging after intratympanic gadolinium (Gd) administration, and to compare it with a conventional 3D-real IR based on the turbo spin echo (TSE) sequence. Methods From July 2021 to November 2022, twenty-seven patients received both the conventional and optimized 3D-real IR sequences after bilateral intratympanic Gd administration. Images of the two sequences were qualitativly evaluated and compared. Contrast-to-noise ratio (CNR), signal-to-noise ratio (SNR), and area ratio of endolymph against the total lymphatic space from the two sequences were measured and compared. Results 14(25.9%) ears with insufficient contrast for the EH diagnosis on the conventional sequence were clearly displayed on the optimized sequence. Image score, CNR and SNR of the optimized sequence were significantly higher than those of the conventional sequence (P < 0.001). The scanning time of two sequences was similar. The area ratio of endolymph against the total lymphatic space in the cochlear was significantly higher on the conventional 3D-real IR than that on the optimized 3D-real IR (P < 0.001); there was no statistical difference in the vestibule between the two sequences. Conclusions Compared with conventional sequence, optimized 3D-real IR sequence with a longer TR may be better for evaluation of EH after intratympanic Gd administration.
5.Application of three-dimensional fluid-attenuated inversion recovery sequence using artificial intelligence-assisted compressed sensing technique in intravenous gadolinium contrast-enhanced magnetic resonance imaging of inner ear
Kai LIU ; Jian WANG ; Huaili JIANG ; Shujie ZHANG ; Di WU ; Xinsheng HUANG ; Mengsu ZENG ; Menglong ZHAO
Chinese Journal of Clinical Medicine 2025;32(2):212-217
Objective To investigate the value of artificial intelligence-assisted compressed sensing (ACS) technology for intravenous gadolinium contrast-enhanced magnetic resonance imaging of the inner ear using three-dimensional fluid-attenuated inversion recovery (3D-FLAIR) sequence. Methods The patients received gadolinium contrast-enhanced magnetic resonance imaging using ACS and united compressed sensing (uCS) 3D-FLAIR at Zhongshan Hospital, Fudan University from January to November 2024 were prospectively enrolled. The repetition time was 16 000 ms, and acquisition time was 6 min 40 s and 10 min 24 s in ACS 3D-FLAIR and uCS 3D-FLAIR, respectively. The images on the two sequences were evaluated independently by two radiologists. The image quality of the two sequences was subjectively evaluated and compared. The signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were compared between the two sequences. The grading consistencies using two sequences and between the two doctors were analyzed. Results There was no statistically difference in subjective score of image quality between the two sequences. SNR and CNR of the ACS 3D-FLAIR sequence were significantly higher than those of the uCS 3D-FLAIR sequence (P<0.001). The kappa values of grades of cochlear and vestibular endolymphatic hydrops were 0.942 and 0.888 using two sequences (P<0.001). The kappa values of grades of cochlear and vestibular endolymphatic hydrops using the ACS 3D-FLAIR sequence between the two doctors were 0.784 and 0.831, respectively (P<0.001); the kappa values of grades of cochlear and vestibular endolymphatic hydrops using uCS 3D-FLAIR sequence between the two doctors were 0.725 and 0.756, respectively (P<0.001). Conclusions ACS 3D-FLAIR could provide higher SNR and CNR than uCS 3D-FLAIR, and is more suitable for intravenous gadolinium contrast-enhanced magnetic resonance imaging of the inner ear; the endolymphatic hydrops grades using ACS 3D-FLAIR is similar to use uCS 3D-FLAIR.
6.Protective effect of Rhein on cerebral ischaemia model mice based on Notch/nuclear factor-κB signaling pathway
Jing-Jing PENG ; Chun-Hua LI ; Kai-Min ZENG ; Ju-Hua GAO
Acta Anatomica Sinica 2025;56(5):541-547
Objective To investigate the neuroprotective effect of rhein(RHE)on ischemic mice and its potential mechanism of reducing inflammatory response and neuronal apoptosis by inhibiting Notch/nuclear factor(NF)-κB signaling pathway.Methods The classical middle cerebral artery occlusion(MCAO)method was used to construct ischemic stroke mouse models.The mice were randomly divided into 5 groups including the sham operation group(sham),the model group(MCAO),the MCAO+edaravone group(Eda),the MCAO+RHE-treated group(RHE),and the MCAO+RHE+Notch activitor Jagged 1 group(RHE+J).Each group has 18 mice.The Bederson scoring system,balance beam walking test and accelerated rotating rod test were used to assess the neurological function and locomotor ability of mice in each group.2,3,5-triphenyltetrazolium chloride(TTC)staining,hematoxylin eosin staining,and TUNEL method were used to assess cerebral infarction,hippocampal morphological damage,and neuronal apoptosis.ELISA was used to analysis the levels of interleukin(IL)-6 and tumor necrosis factor α(TNF-α)in hippocampal tissue.Western blotting was used to analysis caspase-3,Notch 1,Hes1,and NF-κB p65 protein expression.Results Compared with the sham group,Bederson score,balance beam score,cerebral infarct volume,IL-6 and TNF-α levels,the proportion of TUNEL-positively stained cells,caspase-3,Notch1,Hes1,and p-NF-κB p65 protein expression were significantly increased in the MCAO group,whereas the latency to fall decreased significantly(P<0.05).Compared with the MCAO group,Bederson score,balance beam score,cerebral infarct volume,IL-6 and TNF-α levels,proportion of TUNEL-positively stained cells,caspase-3,Notch1,Hes1,and p-NF-κB p65 protein expression were significantly lower in both Eda and RHE groups,whereas the latency to fall increased significantly(P<0.05).Compared with the RHE group,Bederson score,balance beam score,cerebral infarction volume,IL-6 and TNF-α levels,the proportion of TUNEL-positively stained cells,caspase-3,Notch1,Hes1,and p-NF-κB p65 protein expression increased significantly in the RHE+J group,whereas the latency to fall decreased significantly(P<0.05).Conclusion Rhein can significantly improve nerve function in ischemic mice by inhibiting Notch/NF-κB signaling pathway activation,suggesting that rhein has potential clinical application value.
7.Comparative efficacy of Ilizarov ring external fixation and unilateral rail external fixation in the treatment of infected bone defects following surgery for tibial fractures
Kai ZHANG ; Yong LI ; Jia CHANG ; Zhiqiang LIN ; Xiaolong HUANG ; Zequn DENG ; Jian LIU ; Jiangbo HAN ; Fei TAN ; Jiankang ZENG ; Shenghu ZHOU
Chinese Journal of Trauma 2025;41(10):961-968
Objective:To compare the efficacy of Ilizarov ring external fixation and unilateral rail external fixation in the treatment of infected bone defects following surgery for tibial fractures.Methods:A retrospective cohort study was conducted to analyze the clinical data of 50 patients with infected bone defects after surgery for tibial fractures, who were admitted to the 940th Hospital of the Joint Logistics Support Force of the PLA from August 2019 to November 2021, including 37 males and 13 females, aged 19-59 years [(42.2±8.8)years]. After debridement and osteotomy, 28 patients were treated with Ilizarov ring external fixation (Ilizarov group) and 22 with unilateral rail external fixation (unilateral fixation group). All the patients in the two groups had previously undergone internal fixation with plates or Kirschner wires for tibial fracture before bone transport. Bone transport started at one week for three stages after successful infection control and osteotomy and was conducted. The following parameters were compared between the two groups: frame-wearing time and healing index after bone transport, self-rating anxiety scale (SAS) grade at 6 months after bone transport, Paley score and Association for the Study and Application of the Method of Ilizarov (ASAMI) score at the last follow-up, Hospital for Special Surgery (HSS) knee score and Baird-Jackson ankle score on admission, after external fixator removal and at the last follow-up, and incidence of postoperative complications.Results:All the patients were followed up for 28-36 months [(32.5±1.6)months]. There were no significant differences in frame-wearing time or healing index between the two groups after bone transport ( P>0.05). At 6 months after bone transport, the SAS grade in the unilateral fixation group (13 patients with mild anxiety, 8 with moderate anxiety, and 1 with severe anxiety) was better than that in the Ilizarov group (6 patients with mild anxiety, 19 with moderate anxiety, 3 with severe anxiety) ( P<0.01). No significant differences were found in the Paley score or ASAMI score between the two groups at the last follow-up ( P>0.05). There were no significant differences in HSS knee score or Baird-Jackson ankle score between the two groups on admission, after external fixator removal or at the last follow-up ( P>0.05). No significant differences were observed in the incidence of pin tract infection, poor healing, infection in the bone elongation area, or re-fracture between the two groups ( P>0.05). The incidence of postoperative axial deviation was 0 in the Ilizarov group, lower than 18% in the unilateral fixation group (4/22) ( P<0.05). Conclusion:Although Ilizarov ring external fixation and unilateral rail external fixation demonstrate comparable efficacy in the treatment of infected bone defects after surgery for tibial fractures, the former provides superior mechanical stability and postoperative axial deviation correction, while the latter offers advantages in reducing psychological burden and enhancing treatment tolerance.
8.Analysis of scientific and technological outputs of Chinese enterprises manufacturing medical equipment based on bibliometrics:as a case UIH uPMR 790 PET/MRI
Zhiyong LI ; Xiaorui ZHAO ; Kai ZENG ; Zixin KANG ; Han XUE ; Zhan SUN ; Peng NING ; Zeshi CUI ; Xiongjun PENG
China Medical Equipment 2025;22(11):149-163
Objective:To apply data mining and bibliometric methods to collect and analyze literatures related to the uPMR 790 integrated positron emission tomography/magnetic resonance imaging(PET/MRI)of the United Imaging Healthcare(UIH)and assess its output effects in clinical research and application,continued research and development,performance improvement,technology assessment,healthcare technology management,and other aspects.Methods:To conduct literature searches on data platforms such as China National Knowledge Infrastructure(CNKI),WanFang Data Knowledge Service Platform(WANFANG DATA),Wanfang Medical Network(WANFANG MED ONLINE),Chinese Medical Association Journal Database(Yiigle),NIH/PubMed,etc.Based on literature review,to implement bibliometric analysis and evaluate from several technological output dimensions.Results:134 sample literatures were obtained.Among them,there are 121 journal articles,10 dissertations(1 doctoral dissertation and 9 master's theses),3 case reports from the Chinese Medical Case Repository,59 of foreign language journals,and 101 articles sourced from uPMR 790 users as the first author affiliation.Engineering and technical personnel from UIH participated in 58 journal articles;89 clinical research articles,accounting for 66.42%of the sample literatures,15 engineering research and artificial intelligence(AI)research articles,accounting for 11.19%,15 technology assessment articles,accounting for 11.19%,4 healthcare technology management articles,1 experimental research article,and 10 reviews.In addition,33 related patent technologies were retrieved from UIH and its cooperative institutions,as well as UIH PET/MRI equipment users,including 20 invention patents,13 utility model patents.Conclusion:The research results show that the innovative medical equipment,the uPMR 790 PET/MRI,has definite technological outputs in clinical research and application,providing important and widely data of evidence-based medicine for clinical application promotion.It also map out the effective support of this innovative medical equipment for multidisciplinary clinical research such as medical imaging,neurology,cardiovascular and cerebrovascular diseases,geriatrics,pediatrics,etc.Driven by the model of medical-engineering integration and synergy mechanism,display multidimensional output effects such as continuous research and development,quality improvement,technology assessment,configuration management,and healthcare technology management.UIH actively participates in researchs and follows up on innovative practices in medical-engineering integration,demonstrating a positive attitude as a leading medical equipment enterprise.Real world research will provide evidence support for the application demonstration and promotion for equipment manufactured by Chinese medical equipment enterprises.
9.Comparative efficacy of early versus delayed reconstruction of anterior cruciate ligament ruptures under arthroscopy
Jiankang ZENG ; Yingjia ZHOU ; Jiahuan LI ; Fei TAN ; Peijie LI ; Jiangming ZHANG ; Kai ZHANG ; Jing WANG ; Yongjie QIAO ; Shuo YE ; Chenpo DANG ; Shenghu ZHOU
Chinese Journal of Trauma 2025;41(6):572-579
Objective:To compare the efficacy of early versus delayed reconstruction of anterior cruciate ligament (ACL) ruptures under arthroscopy.Methods:A retrospective cohort study was conducted to analyze the clinical data of 260 young adults with ACL ruptures admitted to 940th Hospital of Joint Logistic Support Force of PLA from January 2022 to June 2024, including 171 males and 89 females, aged 18-45 years [(25.6±5.9)years]. Left knee was involved in 127 patients, while the right in 133 patients. All the patients underwent arthroscopic ACL reconstruction with autologous tendon grafts, of whom 130 patients were treated within 3 months after injury (early reconstruction group) but other 130 treated at 3 months after injury (delayed reconstruction group). The operative duration and intraoperative bleeding were compared between the two groups. The incidence of medial meniscus (MM) and lateral meniscus (LM) tears and the incidence of corresponding types of tears were recorded intraoperatively in the two groups. Tegner score and Lysholm score preoperatively, at 3 months postoperatively, and at the last follow-up were detected. Visual analogue scale (VAS) scores at 3 months postoperatively and at the last follow-up and postoperative complication rate were compared between the two groups.Results:All the patients were followed up for 6-15 months [(9.1±3.2)months]. There were no significant differences in operative duration or intraoperative bleeding between the two groups ( P>0.05). The incidence of MM tears in the early reconstruction group was 22.3% (29/130), lower than 34.6% (45/130) in the delayed reconstruction group ( P<0.05). There was no significant difference in the incidence of LM tears between the two groups ( P>0.05). The incidence of MM bucket-handle tears in the early reconstruction group was 2.3% (3/130), lower than 9.2% (12/130) in the delayed reconstruction group ( P<0.05), while no significant difference in the incidence of other types of meniscus tears was found between the two groups ( P>0.05). There were no significant differences in Tegner score or Lysholm score preoperatively between the two groups ( P>0.05). At 3 months postoperatively and at the last follow-up, the Tegner scores in the early reconstruction group were (7.4±1.3)points and (8.6±0.7)points, higher than (6.4±1.5)points and (7.9±0.6)points in the delayed reconstruction group and the Lysholm scores were (82.1±7.1)points and (90.7±3.8)points in the early reconstruction group, higher than (79.5±6.8)points and (86.3±4.0)points in the delayed reconstruction group ( P<0.01). There were no significant differences in VAS scores between the two groups at 3 months postoperatively or at the last follow-up ( P>0.05). The postoperative complication rate was 8.5% (11/130) in the early reconstruction group and 12.3% (16/130) in the delayed reconstruction group ( P>0.05). Conclusion:For young patients with ACL rupture, arthroscopic reconstruction within 3 months after injury can reduce the incidence of MM tear and bucket-handle tear and improve knee function without increasing the incidence of other complications when compared with delayed reconstruction at 3 months after injury.
10.Study on mechanism of naringin in alleviating cerebral ischemia/reperfusion injury based on DRP1/LRRK2/MCU axis.
Kai-Mei TAN ; Hong-Yu ZENG ; Feng QIU ; Yun XIANG ; Zi-Yang ZHOU ; Da-Hua WU ; Chang LEI ; Hong-Qing ZHAO ; Yu-Hong WANG ; Xiu-Li ZHANG
China Journal of Chinese Materia Medica 2025;50(9):2484-2494
This study aims to investigate the molecular mechanism by which naringin alleviates cerebral ischemia/reperfusion(CI/R) injury through DRP1/LRRK2/MCU signaling axis. A total of 60 SD rats were randomly divided into the sham group, the model group, the sodium Danshensu group, and low-, medium-, and high-dose(50, 100, and 200 mg·kg~(-1)) naringin groups, with 10 rats in each group. Except for the sham group, a transient middle cerebral artery occlusion/reperfusion(tMCAO/R) model was established in SD rats using the suture method. Longa 5-point scale was used to assess neurological deficits. 2,3,5-Triphenyl tetrazolium chloride(TTC) staining was used to detect the volume percentage of cerebral infarction in rats. Hematoxylin-eosin(HE) staining and Nissl staining were employed to assess neuronal structural alterations and the number of Nissl bodies in cortex, respectively. Western blot was used to determine the protein expression levels of B-cell lymphoma-2 gene(Bcl-2), Bcl-2-associated X protein(Bax), cleaved cysteine-aspartate protease-3(cleaved caspase-3), mitochondrial calcium uniporter(MCU), microtubule-associated protein 1 light chain 3(LC3), and P62. Mitochondrial structure and autophagy in cortical neurons were observed by transmission electron microscopy. Immunofluorescence assay was used to quantify the fluorescence intensities of MCU and mitochondrial calcium ion, as well as the co-localization of dynamin-related protein 1(DRP1) with leucine-rich repeat kinase 2(LRRK2) and translocase of outer mitochondrial membrane 20(TOMM20) with LC3 in cortical mitochondria. The results showed that compared with the model group, naringin significantly decreased the volume percentage of cerebral infarction and neurological deficit score in tMCAO/R rats, alleviated the structural damage and Nissl body loss of cortical neurons in tMCAO/R rats, inhibited autophagosomes in cortical neurons, and increased the average diameter of cortical mitochondria. The Western blot results showed that compared to the sham group, the model group exhibited increased levels of cleaved caspase-3, Bax, MCU, and the LC3Ⅱ/LC3Ⅰ ratio in the cortex and reduced protein levels of Bcl-2 and P62. However, naringin down-regulated the protein expression of cleaved caspase-3, Bax, MCU and the ratio of LC3Ⅱ/LC3Ⅰ ratio and up-regulated the expression of Bcl-2 and P62 proteins in cortical area. In addition, immunofluorescence analysis showed that compared with the model group, naringin and positive drug treatments significantly decreased the fluorescence intensities of MCU and mitochondrial calcium ion. Meanwhile, the co-localization of DRP1 with LRRK2 and TOMM20 with LC3 in cortical mitochondria was also decreased significantly after the intervention. These findings suggest that naringin can alleviate cortical neuronal damage in tMCAO/R rats by inhibiting DRP1/LRRK2/MCU-mediated mitochondrial fragmentation and the resultant excessive mitophagy.
Animals
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Rats, Sprague-Dawley
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Reperfusion Injury/genetics*
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Flavanones/administration & dosage*
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Rats
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Dynamins/genetics*
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Male
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Brain Ischemia/genetics*
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Protein Serine-Threonine Kinases/genetics*
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Signal Transduction/drug effects*
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Humans
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Drugs, Chinese Herbal/administration & dosage*

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