1.Mechanism of Guihuang Formula in Regulating NLRP3 Inflammasome-mediated Pyroptosis in Treatment of Chronic Prostatitis
Qinghe GAO ; Jianhua FU ; Shengjing LIU ; Ziwei ZHAO ; Ming ZHAO ; Boda GUO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):108-116
ObjectiveTo observe the mechanism of Guihuang formula in regulating the activation of NOD-like receptor protein 3 (NLRP3) inflammasome and inhibiting pyroptosis in the treatment of type Ⅲ prostatitis. Methods(1) In an animal experiment, 50 Sprague Dawley (SD) rats were randomly divided into a blank group, a model group, and low-dose, medium-dose, and high-dose groups of Guihuang formula, with 10 rats in each group. Except for the blank group, the type Ⅲ prostatitis rat model was prepared for the other four groups.After the modeling was successful, the blank group and the model group were given normal saline intragastrically, and the low-dose, medium-dose, and high-dose groups of Guihuang formula were given intragastrically with Guihuang formula (4.9, 9.8, 19.6 g·kg-1). After 30 days of intragastrical administration, samples were taken for detection. Inflammatory cell infiltration in prostate tissue was observed by hematoxylin-eosin (HE) staining, and serum IL-1β and IL-18 levels were measured by enzyme-linked immunosorbent assay (ELISA). Serum malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) levels were determined by biochemistry. NLRP3 expression in prostate tissue was assessed by immunohistochemistry, and the expression of NLRP3, cysteine-aspartic acid protease-1 (Caspase-1), and gasdermin D (GSDMD) in prostate tissue was measured by Western blot. (2) In a cell experiment, human normal prostate epithelial cells (RWPE-1 cells) were divided into a blank group, a model group, a Guihuang formula group, and an NLRP3 inhibitor group (MCC950 group). Except for the blank group, the other three groups were stimulated by 100 μg·L-1 lipopolysaccharide (LPS) for 4 h and 5 mol·L-1 adenosine triphosphate (ATP) for 30 min to prepare the pyroptosis model. After successful modeling, blank serum was given to the blank group and the model group. 6.25 μg·mL-1 Guihuang formula drug-containing serum was added to the Guihuang formula group, and MCC950 was added to the MCC950 group on the basis of the model group. Propidium iodide (PI) uptake and Caspase-1 expression were detected by flow cytometry, and lactate dehydrogenase (LDH) level in the cell supernatant was measured by biochemistry. Interleukin (IL)-1β and IL-18 levels of the cell supernatant were determined by ELISA, and the expression of NLRP3, Caspase-1, and GSDMD was detected in Western blot. Results(1) For the animal experiment, compared with the blank group, the model group showed significant infiltration of inflammatory cells in prostate tissue, while the low-dose, medium-dose, and high-dose groups of Guihuang formula showed reduced infiltration of acinar inflammatory cells, reduced degree of glandular epithelial degeneration and interstitial edema, and significantly reduced degree of damage. Compared with those in the blank group, the levels of IL-1β and IL-18 in the serum of the model group were significantly increased (P<0.01). Compared with the model group, the low-dose, medium-dose, and high-dose groups of Guihuang formula showed a significant decrease in serum IL-1β and IL-18 levels (P<0.01). Compared with that in the blank group, the serum MDA level in the model group significantly increased (P<0.01). Compared with that in the model group, the MDA level in the low-dose, medium-dose, and high-dose groups of Guihuang formula was significantly reduced (P<0.01). Compared with those in the blank group, the levels of SOD and GSH-Px in the serum of the model group significantly decreased (P<0.05). Compared with the model group, the low-dose, medium-dose, and high-dose groups of Guihuang formula showed a significantly increase in SOD (P<0.01). Compared with the model group, the low-dose, medium-dose, and high-dose groups of Guihuang formula showed a significantly increase in GSH-Px (P<0.05). Immunohistochemistry showed that compared with the blank group, the model group had high expression of NLRP3 molecule in prostate tissue. The expression of NLRP3 in the low-dose, medium-dose, and high-dose groups of Guihuang formula was significantly lower than that in the model group. Compared with those in the blank group, the expression levels of NLRP3, Caspase-1, and GSDMD proteins in the prostate tissue of the model group were significantly increased (P<0.01). Compared with those in the model group, the expression levels of NLRP3, Caspase-1, and GSDMD proteins in the low-dose, medium-dose, and high-dose groups of Guihuang formula were significantly inhibited (P<0.01). (2) For the cell experiment, compared with that in the blank group, the PI uptake rate of RWPE-1 cells in the model group significantly increased (P<0.01). Compared with that in the model group, the PI uptake rate of the Guihuang formula group and the inhibitor group significantly decreased (P<0.01). Compared with that in the blank group, the expression of Caspase-1 in the model group was significantly higher (P<0.01). Compared with that in the model group, the Caspase-1 in the Guihuang formula group and the inhibitor group significantly decreased (P<0.01). Compared with the blank group, the model group showed an increase in LDH release (P<0.01). Compared with the model group, the Guihuang formula group and the inhibitor group showed a significantly decrease in LDH release (P<0.01). Compared with those in the blank group, the levels of IL-1β and IL-18 in the supernatant of the model group were significantly increased (P<0.01). Compared with the model group, the Guihuang formula group and the inhibitor group showed a significantly decrease in the levels of IL-1β and IL-18 (P<0.01). Compared with those in the blank group, the expression levels of NLRP3, Caspase-1, and GSDMD proteins significantly increased in the model group (P<0.01). Compared with those in the model group, the protein expression levels of NLRP3, Caspase-1, and GSDMD were significantly reduced in the Guihuang formula group and inhibitor group (P<0.01). ConclusionGuihuang formula can inhibit the activation of Caspase-1, prevent GSDMD cleavation and lysis, and inhibit cell pyrodeath in the treatment of type Ⅲ prostatitis by inhibiting the activation of NLRP3 inflammasome.
2.Study on the effects and mechanisms of Lycium ruthenicum Murr. in improving sleep
Ming QIAO ; Yao ZHAO ; Yi ZHU ; Yexia CAO ; Limei WEN ; Yuehong GONG ; Xiang LI ; Juanchen WANG ; Tao WANG ; Jianhua YANG ; Junping HU
China Pharmacy 2026;37(1):24-29
OBJECTIVE To investigate the effects and mechanisms of Lycium ruthenicum Murr. in improving sleep. METHODS Network pharmacology was employed to identify the active components of L. ruthenicum and their associated disease targets, followed by enrichment analysis. A caffeine‑induced zebrafish model of sleep deprivation was established , and the zebrafish were treated with L. ruthenicum Murr. extract (LRME) at concentrations of 0.1, 0.2 and 0.4 mg/mL, respectively; 24 h later, behavioral changes of zebrafish and pathological alterations in brain neurons were subsequently observed. The levels of inflammatory factors [interleukin-6 (IL-6), IL-1β, IL-10, tumor necrosis factor-α (TNF-α)], oxidative stress markers [superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), catalase (CAT)], and neurotransmitters [5- hydroxytryptamine (5-HT), γ-aminobutyric acid (GABA), glutamic acid (Glu), dopamine (DA), and norepinephrine (NE)] were measured. The protein expression levels of protein kinase B1 (AKT1), phosphorylated AKT1 (p-AKT1), epidermal growth factor receptor (EGFR), B-cell lymphoma 2 (Bcl-2), sarcoma proto-oncogene,non-receptor tyrosine kinase (SRC), and heat shock protein 90α family class A member 1 (HSP90AA1) in the zebrafish were also determined. RESULTS A total of 12 active components and 176 intersecting disease targets were identified through network pharmacology analysis. Among these, apigenin, naringenin and others were recognized as core active compounds, while AKT1, EGFR and others served as key targets; EGFR tyrosine kinase inhibitor resistance signaling pathway was identified as the critical pathway. The sleep improvement rates in zebrafish of LRME low-, medium-, and high-dose groups were 54.60%, 69.03% and 77.97%, 开发。E-mail:hjp_yft@163.com respectively, while the inhibition ratios of locomotor distance were 0.57, 0.83 and 0.95, respectively. Compared with the model group, the number of resting counts, resting time and resting distance were significantly increased/extended in LRME medium- and high-dose groups (P<0.05). Neuronal damage in the brain was alleviated. Additionally, the levels of IL-6, IL-1β, TNF-α, MDA, Glu, DA and NE, as well as the protein expression levels of AKT1, p-AKT1, EGFR, SRC and HSP90AA1, were markedly reduced (P<0.05), while the levels of IL-10, SOD, GSH-Px, CAT, 5-HT and GABA, as well as Bcl-2 protein expression, were significantly elevated (P<0.05). CONCLUSIONS L. ruthenicum Murr. demonstrates sleep-improving effects, and its specific mechanism may be related to the regulation of inflammatory responses, oxidative stress, neurotransmitter balance, and the EGFR tyrosine kinase inhibitor resistance signaling pathway.
3.Repair strategies for nonunion in old osteoporotic vertebral compression fractures:a case analysis
Xingcheng ZHAO ; Jun WANG ; Ming LU
Chinese Journal of Tissue Engineering Research 2025;29(3):538-546
BACKGROUND:Patients with osteoporotic compression fractures of the lumbar spine have varying degrees of changes in the sagittal parameters of the spine and pelvis,resulting in disequilibrium and rebalancing of the sagittal plane of the whole spine/lumbar spine.If the primary osteoporotic vertebral compression fracture is not treated promptly,it develops into an old osteoporotic vertebral compression fracture with nonunion over time. OBJECTIVE:To observe the effect of percutaneous vertebroplasty in nine patients with old osteoporotic lumbar compression fracture with nonunion. METHODS:A retrospective analysis was conducted on the clinical data of nine elderly patients with old osteoporotic lumbar compression fractures with nonunion treated in the Orthopedics Department of Shanghai Public Health Clinical Center from September 2018 to August 2022.All patients underwent revision treatment with percutaneous vertebroplasty and received regular anti-osteoporosis treatment and rehabilitation training.The lumbar spine X-ray plain film and three-dimensional CT test results were reviewed before discharge and during follow-up to evaluate the diffusion of bone cement.Visual analog scale score,lumbar Oswestry disability index score,and Roussouly classification were recorded before,after surgery and during follow-up. RESULTS AND CONCLUSION:(1)During postoperative follow-up,none of the 9 cases in this group experienced serious surgical complications such as nerve injury,infection,bone cement leakage,pulmonary embolism,and allergic shock,and no adverse reactions related to bone cement occurred.One patient died of respiratory failure due to COVID-19 infection 2 years after operation.(2)The visual analog scale score and Oswestry disability index score of postoperative pain in patients with recurrent vertebral fractures after percutaneous vertebroplasty were significantly lower than those before revision surgery.After the last follow-up percutaneous vertebroplasty revision surgery,the Roussouly classification of patients significantly improved.There was no statistically significant difference between the changes in vertebral body height during the last follow-up measurement and preoperative comparison.(3)These findings indicate that for patients with old osteoporotic lumbar compression fractures with nonunion,moderate injection of bone cement and effective dispersion of bone cement have a significant relieving effect on postoperative pain.Regular anti-osteoporosis and effective postoperative rehabilitation training can help patients improve their postoperative lumbar spine function.
4.Research and Application of Scalp Surface Laplacian Technique
Rui-Xin LUO ; Si-Ying GUO ; Xin-Yi LI ; Yu-He ZHAO ; Chun-Hou ZHENG ; Min-Peng XU ; Dong MING
Progress in Biochemistry and Biophysics 2025;52(2):425-438
Electroencephalogram (EEG) is a non-invasive, high temporal-resolution technique for monitoring brain activity. However, affected by the volume conduction effect, EEG has a low spatial resolution and is difficult to locate brain neuronal activity precisely. The surface Laplacian (SL) technique obtains the Laplacian EEG (LEEG) by estimating the second-order spatial derivative of the scalp potential. LEEG can reflect the radial current activity under the scalp, with positive values indicating current flow from the brain to the scalp (“source”) and negative values indicating current flow from the scalp to the brain (“sink”). It attenuates signals from volume conduction, effectively improving the spatial resolution of EEG, and is expected to contribute to breakthroughs in neural engineering. This paper provides a systematic overview of the principles and development of SL technology. Currently, there are two implementation paths for SL technology: current source density algorithms (CSD) and concentric ring electrodes (CRE). CSD performs the Laplace transform of the EEG signals acquired by conventional disc electrodes to indirectly estimate the LEEG. It can be mainly classified into local methods, global methods, and realistic Laplacian methods. The global method is the most commonly used approach in CSD, which can achieve more accurate estimation compared with the local method, and it does not require additional imaging equipment compared with the realistic Laplacian method. CRE employs new concentric ring electrodes instead of the traditional disc electrodes, and measures the LEEG directly by differential acquisition of the multi-ring signals. Depending on the structure, it can be divided into bipolar CRE, quasi-bipolar CRE, tripolar CRE, and multi-pole CRE. The tripolar CRE is widely used due to its optimal detection performance. While ensuring the quality of signal acquisition, the complexity of its preamplifier is relatively acceptable. Here, this paper introduces the study of the SL technique in resting rhythms, visual-related potentials, movement-related potentials, and sensorimotor rhythms. These studies demonstrate that SL technology can improve signal quality and enhance signal characteristics, confirming its potential applications in neuroscientific research, disease diagnosis, visual pathway detection, and brain-computer interfaces. CSD is frequently utilized in applications such as neuroscientific research and disease detection, where high-precision estimation of LEEG is required. And CRE tends to be used in brain-computer interfaces, that have stringent requirements for real-time data processing. Finally, this paper summarizes the strengths and weaknesses of SL technology and envisages its future development. SL technology boasts advantages such as reference independence, high spatial resolution, high temporal resolution, enhanced source connectivity analysis, and noise suppression. However, it also has shortcomings that can be further improved. Theoretically, simulation experiments should be conducted to investigate the theoretical characteristics of SL technology. For CSD methods, the algorithm needs to be optimized to improve the precision of LEEG estimation, reduce dependence on the number of channels, and decrease computational complexity and time consumption. For CRE methods, the electrodes need to be designed with appropriate structures and sizes, and the low-noise, high common-mode rejection ratio preamplifier should be developed. We hope that this paper can promote the in-depth research and wide application of SL technology.
5.Effects of Exercise Training on The Behaviors and HPA Axis in Autism Spectrum Disorder Rats Through The Gut Microbiota
Xue-Mei CHEN ; Yin-Hua LI ; Jiu-Gen ZHONG ; Zhao-Ming YANG ; Xiao-Hui HOU
Progress in Biochemistry and Biophysics 2025;52(6):1511-1528
ObjectiveThe study explores the influence of voluntary wheel running on the behavioral abnormalities and the activation state of the hypothalamic-pituitary-adrenal (HPA) axis in autism spectrum disorder (ASD) rats through gut microbiota. MethodsSD female rats were selected and administered either400 mg/kg of valproic acid (VPA) solution or an equivalent volume of saline via intraperitoneal injection on day 12.5 of pregnancy. The resulting offspring were divided into 2 groups: the ASD model group (PASD, n=35) and the normal control group (PCON, n=16). Behavioral assessments, including the three-chamber social test, open field test, and Morris water maze, were conducted on postnatal day 23. After behavioral testing, 8 rats from each group (PCON, PASD) were randomly selected for serum analysis using enzyme-linked immunosorbent assay (ELISA) to measure corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH), and corticosterone (CORT) concentration, to evaluate the functional state of the HPA axis in rats. On postnatal day 28, the remaining 8 rats in the PCON group were designated as the control group (CON, n=8), and the remaining 27 rats in the PASD group were randomly divided into 4 groups: ASD non-intervention group (ASD, n=6), ASD exercise group (ASDE, n=8), ASD fecal microbiota transplantation group (FMT, n=8), and ASD sham fecal microbiota transplantation group (sFMT, n=5). The rats in the ASD group and the CON group were kept under standard conditions, while the rats in the ASDE group performed 6 weeks of voluntary wheel running intervention starting on postnatal day 28. The rats in the FMT group were gavaged daily from postnatal day 42 with 1 ml/100 g fresh fecal suspension from ASDE rats which had undergone exercise for 2 weeks, 5 d per week, continuing for 4 weeks. The sFMT group received an equivalent volume of saline. After the interventions were completed, behavioral assessments and HPA axis markers were measured for all groups. ResultsBefore the intervention, the ASD model group exhibited significantly reduced social ability, social novelty preference, spontaneous activity, and exploratory interest, as well as impaired spatial learning, memory, and navigation abilities compared to the normal control group (P<0.05). Serum concentration of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH), and corticosterone (CORT) in the PASD group were significantly higher than those in the PCON group (P<0.05). Following 6 weeks of voluntary wheel running, the ASDE group showed significant improvements in social ability, social novelty preference, spontaneous activity, exploratory interest, spatial learning, memory, and navigation skills compared to the ASD group (P<0.05), with a significant decrease in serum CORT concentration (P<0.05), and a downward trend in CRH and ACTH concentration. After 4 weeks of fecal microbiota transplantation in the exercise group, the FMT group showed marked improvements in social ability, social novelty preference, spontaneous activity, exploratory interest, as well as spatial learning, memory, and navigation abilities compared to both the ASD and sFMT groups (P<0.05). In addition, serum ACTH and CORT concentration were significantly reduced (P<0.05), and CRH concentration also showed a decreasing trend. ConclusionExercise may improve ASD-related behaviors by suppressing the activation of the HPA axis, with the gut microbiota likely playing a crucial role in this process.
6.The correlation between abnormal metabolic indexes and the severity of coronary artery lesions in patients with acute coronary syndrome
Yajun ZHAO ; Ming LIU ; Yuxiang DAI ; Xiaopan LI ; Xuelin CHENG ; Qizhe WANG ; Ru LIU ; Yaxin XU ; Sunfang JIANG
Chinese Journal of Clinical Medicine 2025;32(3):441-448
Objective To explore the influencing factors of coronary artery lesion severity in patients with acute coronary syndrome (ACS). Methods Clinical data of ACS patients admitted to Zhongshan Hospital, Fudan University from December 2017 to December 2019 were consecutively collected. The modified Gensini score was used to assess the severity of coronary artery lesions. Univariate and multivariate linear regression analyses were performed to identify independent factors associated with coronary artery lesion severity. Results A total of 1 689 ACS patients were included, with an average age of (64.04±11.45) years; 1 353 (80.11%) were male, and the mean modified Gensini score was (8.12±4.03). Multivariate linear regression analysis revealed that sex (β=0.97, P=0.001), age (β=0.03, P=0.021), estimated glomerular filtration rate (eGFR; β=-0.03, P<0.001), low-density lipoprotein cholesterol (LDL-C; β=0.58, P<0.001), apolipoprotein A1 (Apo A1; β=-1.28, P=0.012), lipoprotein(a) [Lp(a); β=0.001, P=0.033], and glycated hemoglobin A1C (HbA1C; β=0.45, P<0.001) were independent influencing factors of the modified Gensini score. Conclusions Metabolic indicators, including Apo A1, LDL-C, HbA1C, and Lp(a), may serve as risk factors for coronary artery lesion severity in ACS patients, with Apo A1 demonstrating the strongest impact.
7.Efficacy and safety of robot-assisted modified Y-shaped ileal orthotopic neobladder reconstruction
Tonglei ZHAO ; Weipu MAO ; Yiduo WANG ; Bin XU ; Shuqiu CHEN ; Weidong ZHU ; Ming CHEN ; Jianping WU
Journal of Modern Urology 2025;30(2):137-142
Objective: To investigate the efficacy and safety of robot-assisted modified Y-shaped ileal orthotopic neobladder reconstruction,so as to provide reference for clinical practice. Methods: The clinical data of 44 patients who underwent robot-assisted laparoscopic radical cystectomy,lymph node dissection,and modified Y-shaped ileal orthotopic neobladder reconstruction during Feb.2020 and Aug.2022 were retrospectively analyzed.The surgical position,Trocar position,and key surgical steps were reported.The perioperative conditions,postoperative complications,neobladder volume,maximum urinary flow rate,postvoid residual,renal function,and urinary control function were recorded. Results: All 44 surgeries were successfully completed,with operation time of (314.32±51.02) min,modified Y-shaped ileal orthotopic neobladder reconstruction time of (103.52±9.56) min,and bleeding volume of (128.18±57.27) mL.The postoperative time for fluid intake was (4.16±0.86) days,catheter indwelling time was (14.02±3.20) days,and patients were discharged 1 to 2 days after catheter removal.Clavien-Dindo grade Ⅱ and Ⅲ complications occurred in 15 and 2 patients,respectively.During the follow-up of (20.77±5.90) months,dysuria occurred in 1 case,urethral calculi in 2 cases,and incomplete bowel obstruction in 2 cases. The postoperative neobladder capacity was (195.75±15.51) mL,maximal urinary flow rate (20.30±2.05) mL/s,postvoid residual (19.86±13.80) mL and serum creatinine (81.98±25.97) μmol/L. The incidence of daytime and nocturnal urinary incontinence 3,6 and 12 months after operation were 20.45% and 29.55%,11.36% and 18.18%,and 4.55% and 9.09%,respectively. Conclusion: Robot-assisted modified Y-shaped ileal orthotopic neobladder reconstruction has favorable efficacy and safety,and low incidence of postoperative complications,which can be applied in clinical practice.
8.Clinical observation of Huangkui capsule combined with cyclophosphamide and prednisone in the treatment of IgA nephropathy with renal insufficiency
Yan JIANG ; Ming ZHANG ; Guoxin LIU ; Chenyang ZHAO
China Pharmacy 2025;36(15):1899-1903
OBJECTIVE To explore the clinical efficacy and safety of Huangkui capsule combined with cyclophosphamide and prednisone in the treatment of immunoglobulin A (IgA) nephropathy with renal insufficiency. METHODS A total of 117 patients with IgA nephropathy and renal insufficiency who were hospitalized in the department of nephrology of the Second Affiliated Hospital of Dalian Medical University from February 2021 to March 2024 were divided into prednisone group (n=38), cyclophosphamide group (n=39) and Huangkui group (n=40) according to the random number table method. On the basis of standardized basic treatment, the three groups were treated with prednisone, prednisone + cyclophosphamide, and prednisone + cyclophosphamide + Huangkui capsule, respectively, with a course of 6 months. The clinical efficacy, renal function indexes, immunoglobulin levels, inflammatory factor levels before and after treatment, and the incidence of adverse reactions during treatment were compared among the three groups. RESULTS Finally, 107 patients completed the study (35 in prednisone group, 37 in cyclophosphamide group, and 35 in Huangkui group). After 6 months of treatment, there was a statistically significant difference in the total effective rate among the three groups (P=0.028), and the total effective rate of the Huangkui group was significantly higher than that of the prednisone group (P=0.023). In terms of renal function, the levels of blood urea nitrogen (BUN), serum creatinine (Scr), and urinary microalbumin (Umalb) in the three groups after treatment were significantly lower than those before treatment, while the estimated glomerular filtration rate (eGFR) was significantly higher than that before treatment (P<0.05). Among them, the Huangkui group was superior to the other two groups in reducing BUN level (P<0.05), and both the Huangkui group and cyclophosphamide group were superior to the prednisone group in improving Scr, Umalb and eGFR (P<0.05). In terms of immunology, both the Huangkui group and cyclophosphamide group were superior to the prednisone group in increasing IgG level and decreasing IgA and IgM levels (P<0.05). In terms of inflammatory factors, E-mail:amychina0411@163.com the Huangkui group was superior to the prednisone group and cyclophosphamide group in reducing tumor necrosis factor-α level (P<0.05), and superior to the prednisone group in reducing interleukin-6 level (P<0.05). There was no statistically significant difference in the total incidence of adverse reactions among the three groups (P>0.05). CONCLUSIONS Huangkui capsule combined with cyclophosphamide and prednisone has a good therapeutic effect on IgA nephropathy with renal insufficiency. It can further improve patients’ renal function and immune function, regulate inflammatory status, and has good safety.
9.The Functional Diversity and Regulatory Mechanism of Clathrin Plaques
Yi-Ge ZHAO ; Zhao-Hong JIANG ; Qian-Yi ZHOU ; Zhi-Ming CHEN
Progress in Biochemistry and Biophysics 2025;52(8):1958-1971
Clathrin-mediated endocytosis (CME) is a critical process by which cells internalize macromolecular substances and initiate vesicle trafficking, serving as the foundation for many cellular activities. Central to this process are clathrin-coated structures (CCSs), which consist of clathrin-coated pits (CCPs) and clathrin plaques. While clathrin-coated pits are well-established in the study of endocytosis, clathrin plaques represent a more recently discovered but equally important component of this system. These plaques are large, flat, and extended clathrin-coated assemblies found on the cytoplasmic membrane. They are distinct from the more typical clathrin-coated pits in terms of their morphology, larger surface area, and longer lifespan. Recent research has revealed that clathrin plaques play roles that go far beyond endocytosis, contributing to diverse cellular processes such as cellular adhesion, mechanosensing, migration, and pathogen invasion. Unlike traditional clathrin-coated pits, which are transient and dynamic structures involved primarily in the internalization of molecules, clathrin plaques are more stable and extensive, often persisting for extended periods. Their extended lifespan suggests that they serve functions beyond the typical endocytic role, making them integral to various cellular processes. For instance, clathrin plaques are involved in the regulation of intercellular adhesion, allowing cells to better adhere to one another or to the extracellular matrix, which is crucial for tissue formation and maintenance. Furthermore, clathrin plaques act as mechanosensitive hubs, enabling the cell to sense and respond to mechanical stress, a feature that is essential for processes like migration, tissue remodeling, and even cancer progression. Recent discoveries have also highlighted the role of clathrin plaques in cellular signaling. These plaques can serve as scaffolds for signaling molecules, orchestrating the activation of various pathways that govern cellular behavior. For example, the recruitment of actin-binding proteins such as F-actin and vinculin to clathrin plaques can influence cytoskeletal dynamics, helping cells adapt to mechanical changes in their environment. This recruitment also plays a pivotal role in regulating cellular migration, which is crucial for developmental processes. Additionally, clathrin plaques influence receptor-mediated signal transduction by acting as platforms for the assembly of signaling complexes, thereby affecting processes such as growth factor signaling and cellular responses to extracellular stimuli. Despite the growing body of evidence that supports the involvement of clathrin plaques in a wide array of cellular functions, much remains unknown about the precise molecular mechanisms that govern their formation, maintenance, and turnover. For example, the factors that regulate the recruitment of clathrin and other coat proteins to form plaques, as well as the signaling molecules that coordinate plaque dynamics, remain areas of active research. Furthermore, the complex interplay between clathrin plaques and other cellular systems, such as the actin cytoskeleton and integrin-based adhesion complexes, needs further exploration. Studies have shown that clathrin plaques can respond to mechanical forces, with recent findings indicating that they act as mechanosensitive structures that help the cell adapt to changing mechanical environments. This ability underscores the multifunctional nature of clathrin plaques, which, in addition to their role in endocytosis, are involved in cellular processes such as mechanotransduction and adhesion signaling. In summary, clathrin plaques represent a dynamic and versatile component of clathrin-mediated endocytosis. They play an integral role not only in the internalization of macromolecular cargo but also in regulating cellular adhesion, migration, and signal transduction. While much has been learned about their structural and functional properties, significant questions remain regarding the molecular mechanisms that regulate their formation and their broader role in cellular physiology. This review highlights the evolving understanding of clathrin plaques, emphasizing their importance in both endocytosis and a wide range of other cellular functions. Future research is needed to fully elucidate the mechanisms by which clathrin plaques contribute to cellular processes and to better understand their implications for diseases, including cancer and tissue remodeling. Ultimately, clathrin plaques are emerging as crucial hubs that integrate mechanical, biochemical, and signaling inputs, providing new insights into cellular function and the regulation of complex cellular behaviors.
10.Preparation and evaluation of long-acting light-protective nanogel based on fullerenol-cerium oxide composite system
Tianlong ZHANG ; Jia LIU ; Qing ZHAO ; Yue ZHOU ; Ming YANG ; Qianyu LUO
China Pharmacy 2025;36(17):2106-2112
OBJECTIVE To develop a long-acting light-protective nanogel with both physical barrier and chemical clearance functions, and evaluate its performance. METHODS The photoprotective nanogel composed of mussel mucin and sodium hyaluronate was constructed based on a fullerenol-cerium oxide composite nano system, namely fullerenol-cerium oxide nanogel (FCN), and was characterized. The antioxidant capacity of FCN was evaluated using in vitro free radical scavenging experiments; its UV shielding ability was assessed by using an SPF value detector; its biosafety was assessed according to the requirements of the Guidelines for Drug Safety Evaluation; skin adhesion was assessed using small animal 3D live imaging technology; its sun protection ability was assessed through skin sunscreen detection and histopathological observation. RESULTS The average particle sizes of cerium oxide and fullerenol nanoparticles in FCN were about 20 and 10 nm, respectively, and FCN exhibited good UV absorption and free radical scavenging abilities. SPF value of FCN was 58.95±0.82, and the ultraviolet A protection level value was 6.21±0.15. No pathogenic colonies such as Staphylococcus aureus, were detected in the nanogel, and the contents of lead, arsenic, mercury and cadmium all met the standards for pharmaceutical excipients; FCN group did not show any irritating reactions such as erythema, edema, or desquamation; blood biochemical indicators of the FCN group were within the normal reference range. The material clearance rate of mice in the artificial sweat flushing group was less than 30%, while the material clearance rate of mice in the dry cleaning group reached about 92%. The mice in the protective group did not show obvious erythema or ulcer formation throughout the experiment. Histopathology showed that the fibers were arranged in an orderly manner, and the number of collagen fibers was close to that of the control group. CONCLUSIONS The FCN formulation constructed in this study meets the relevant requirements of the Chinese Pharmacopoeia, has good safety and skin compatibility, and achieves dual synergistic protection of UV shielding and free radical scavenging.

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