1.Mechanism of Huayu Jiedu Prescription in Preventing and Treating Cerebral Ischemia Injury by Regulating NETosis After Acute Cerebral Infarction with Blood Stasis and Toxin Syndrome
Wuchaonan LIU ; Dingxiang LI ; Le YANG ; Jing LIU ; Shengping LUO ; Fang LEI ; Hanlin LEI ; Yihui DENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):50-59
ObjectiveTo investigate the material basis of the pathogenesis of cerebral ischemic injury with blood stasis and toxin syndrome and to explore the protective effects of Huayu Jiedu prescription (HYJDP) on neutrophil extracellular trap-related cell death (NETosis) in cerebral ischemic injury following acute cerebral infarction. MethodsSeventy-two Sprague-Dawley (SD) rats were randomly divided into six groups (n=12 per group): sham operation (Sham) group, blood stasis and toxin model (Model) group, low-, medium-, and high-dose HYJDP groups (HYJDP-L, HYJDP-M, and HYJDP-H; 9, 18, and 36 g·kg-1, respectively), and butylphthalide (NBP) group (0.06 g·kg-1). Except for the Sham group, rats in all other groups were subjected to carrageenan/dry yeast combined with a modified intraluminal filament method to establish a focal cerebral ischemia model of the middle cerebral artery with blood stasis and toxin syndrome. Neurological function was evaluated at 24 h after modeling using the Zea-Longa neurological deficit score. Cerebral infarction rate was assessed by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Pathological morphology of brain tissue was observed using hematoxylin-eosin (HE) staining. Enzyme-linked immunosorbent assay (ELISA) was used to determine serum levels of interleukin-8 (IL-8), myeloperoxidase-DNA complexes (MPO-DNA), and citrullinated histone H3 (CitH3). Protein expression of phosphorylated phosphatidylinositol 3-kinase (p-PI3K), protein kinase B (p-Akt), mammalian target of rapamycin (p-mTOR), sequestosome 1 (p62), and CitH3 in brain tissue was detected by Western blot. Immunofluorescence (IF) was used to detect the expression of neutrophil-specific marker Ly6G, CitH3, and neuron-specific nuclear protein (NeuN) in brain tissue. ResultsCompared with the Sham group, neurological deficit scores and cerebral infarction rates in the model group were significantly increased (P<0.01 for both). HE staining showed varying degrees of neuronal degeneration and necrosis, characterized by blurred neuronal structures, nuclear pyknosis and fragmentation, cytoplasmic dissolution into a vacuolated reticular pattern, and mild glial cell proliferation. ELISA results showed that serum levels of IL-8, MPO-DNA, and CitH3 were significantly increased (P<0.01). Western blot analysis demonstrated decreased expression of p-PI3K, p-Akt, p-mTOR, and p62, while CitH3 expression was significantly increased (P<0.01). IF results showed an increased number of NETs+ cells and a significant decrease in NeuN+ cells (P<0.01). Compared with the Model group, neurological deficit scores in the HYJDP-H group were significantly decreased (P<0.05), and cerebral infarction rates in the HYJDP-H and NBP groups were significantly reduced (P<0.01). HE staining showed that brain tissue damage was markedly alleviated in the HYJDP-H group. ELISA results showed that levels of IL-8, MPO-DNA, and CitH3 were significantly decreased in the HYJDP-M, HYJDP-H, and NBP groups (P<0.01). Western blot analysis showed that expression of p-PI3K, p-Akt, p-mTOR, and p62 was significantly increased in the HYJDP-H and NBP groups, while CitH3 expression was significantly reduced in all drug intervention groups (P<0.01). IF results showed that the number of NETs+ cells was significantly decreased and the number of NeuN⁺ cells was significantly increased in all drug intervention groups (P<0.01). ConclusionNETs may be the material basis of the pathogenesis of cerebral ischemic injury characterized by blood stasis and toxin. HYJDP can regulate the PI3K/Akt/mTOR signaling pathway, reduce the release of pro-inflammatory mediators and NETosis-related products, alleviate cerebral ischemic injury caused by autophagy-dependent NETosis, and thereby exert a neuroprotective effect.
2.Coupling of an Au@AgPt nanozyme array with an micrococcal nuclease-specific responsiveness strategy for colorimetric/SERS sensing of Staphylococcus aureus in patients with sepsis
Xueqin HUANG ; Yingqi YANG ; Hanlin ZHOU ; Liping HU ; Annan YANG ; Hua JIN ; Biying ZHENG ; Jiang PI ; Jun XU ; Pinghua SUN ; Huai-Hong CAI ; Xujing LIANG ; Bin PAN ; Junxia ZHENG ; Haibo ZHOU
Journal of Pharmaceutical Analysis 2025;15(2):389-400
Rapid and ultrasensitive detection of pathogen-associated biomarkers is vital for the early diagnosis and therapy of bacterial infections.Herein,we developed a close-packed and ordered Au@AgPt array coupled with a cascade triggering strategy for surface-enhanced Raman scattering(SERS)and colorimetric identification of the Staphylococcus aureus biomarker micrococcal nuclease(MNase)in serum samples.The trimetallic Au@AgPt nanozymes can catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine(TMB)molecules to SERS-enhanced oxidized TMB(oxTMB),accompanied by the color change from colorless to blue.In the presence of S.aureus,the secreted MNase preferentially cut the nucleobase AT-rich regions of DNA sequences on magnetic beads(MBs)to release alkaline phosphatase(ALP),which subsequently mediated the oxTMB reduction for inducing the colorimetric/SERS signal fade away.Using this"on-to-off"triggering strategy,the target S.aureus can be recorded in a wide linear range with a limit of detection of 38 CFU/mL in the colorimetric mode and 6 CFU/mL in the SERS mode.Meanwhile,the MNase-mediated strategy characterized by high specificity and sensitivity successfully discriminated between patients with sepsis(n=7)and healthy participants(n=3),as well as monitored the prog-nostic progression of the disease(n=2).Overall,benefiting from highly active and dense"hot spot"substrate,MNase-mediated cascade response strategy,and colorimetric/SERS dual-signal output,this methodology will offer a promising avenue for the early diagnosis of S.aureus infection.
3.Mechanism of Fas-associated protein with death domain in promoting proliferation of head and neck squamous cell carcinoma cells
Yinan CHEN ; Yang ZHENG ; Hanlin ZENG ; Ming LEI
Journal of Shanghai Jiaotong University(Medical Science) 2025;45(4):404-414
Objective·To detect the expression level of Fas-associated protein with death domain(FADD)in head and neck squamous cell carcinoma(HNSCC)and to explore the molecular mechanisms by which FADD promotes the proliferation of HNSCC cells.Methods·The GEPIA 2 database was utilized to analyze the expression level of FADD in tumor tissues and to evaluate its association with prognosis.Immunohistochemistry staining(IHC)was performed on HNSCC tissues to investigate the changes in FADD expression levels in normal,dysplastic,and tumor tissues.Stable FADD-knockdown Fadu and HSC3 cell lines were constructed and validated using Western blotting and quantitative real-time PCR(qRT-PCR).The regulatory effect of FADD on the proliferation of HNSCC cells was explored using the LiveCyte live-cell tracking system,colony formation assay,and cell viability assay.Proteins interacting with FADD were identified by co-immunoprecipitation mass spectrometry(Co-IP/MS),and further mechanistic studies were conducted using CRISPR/Cas9 technology,LiveCyte live-cell tracking system,and Western blotting.Results·Analysis of the GEPIA2 database indicated that FADD was significantly overexpressed in head and neck cancer and was associated with poor prognosis.IHC staining showed that FADD expression levels progressively increased from normal to dysplastic to tumor tissues in HNSCC patients.Knockdown of FADD in HNSCC cells resulted in significantly reduced proliferation and colony formation compared to the control group.Co-IP/MS results showed that FADD interacted with the CUX1 protein,and FADD knockdown led to increased CUX1 expression.Moreover,CUX1 knockdown significantly promoted HNSCC cell proliferation and reversed the anti-proliferative phenotype caused by FADD knockdown.Conclusion·FADD plays a significant pro-carcinogenic role in HNSCC and is associated with poor prognosis.FADD can further regulate tumor cell proliferation by interacting with CUX1 and suppressing its expression level.
4.Application of infrared thermography in auxiliary diagnosis of neck and lumbar spine training injuries in aircraft pilots
Baifeng SUN ; Hanlin SONG ; Xiao JIANG ; Zichuan WU ; Aochen XU ; Shihao LU ; Yang LIU
Journal of Navy Medicine 2025;46(6):543-546
Objective To explore the differences in neck and lumbar spine injuries among different types of aircraft pilots and the correlation between body surface temperature and the severity of injury.Methods Data were collected by questionnaire surveys and medical examinations.Forty male pilots were selected as research objects,including 14 fighter pilots and 26 helicopter pilots,aged between 28 and 39 years,with a height range of 168 to 178 cm,and a total flight time of 600 to 2 000 h.Infrared thermal imaging was employed for skin temperature.A questionnaire survey was conducted for the assessment of the common site and degree of pain.The relationship between body surface temperature and pain was investigated.Results Fighter pilots mainly complained of discomfort in the neck and scapular region,while helicopter pilots were more likely to experience lower back pain.The skin temperature in the affected regions was significantly higher than that in the surrounding normal regions in both fighter pilots and helicopter pilots(P<0.05).The pain score was positively related with elevated temperature in the affected areas(P<0.05).Conclusion It is found that fighter pilots are more prone to neck and scapular pain,while helicopter pilots are more likely to experience lower back pain.The severity of pain is positively related with increased temperature in the affected areas.Infrared thermal imaging can be used to assess temperature variation at injured sites and the location of training injuries among pilots.
5.Mechanism of Fas-associated protein with death domain in promoting proliferation of head and neck squamous cell carcinoma cells
Yinan CHEN ; Yang ZHENG ; Hanlin ZENG ; Ming LEI
Journal of Shanghai Jiaotong University(Medical Science) 2025;45(4):404-414
Objective·To detect the expression level of Fas-associated protein with death domain(FADD)in head and neck squamous cell carcinoma(HNSCC)and to explore the molecular mechanisms by which FADD promotes the proliferation of HNSCC cells.Methods·The GEPIA 2 database was utilized to analyze the expression level of FADD in tumor tissues and to evaluate its association with prognosis.Immunohistochemistry staining(IHC)was performed on HNSCC tissues to investigate the changes in FADD expression levels in normal,dysplastic,and tumor tissues.Stable FADD-knockdown Fadu and HSC3 cell lines were constructed and validated using Western blotting and quantitative real-time PCR(qRT-PCR).The regulatory effect of FADD on the proliferation of HNSCC cells was explored using the LiveCyte live-cell tracking system,colony formation assay,and cell viability assay.Proteins interacting with FADD were identified by co-immunoprecipitation mass spectrometry(Co-IP/MS),and further mechanistic studies were conducted using CRISPR/Cas9 technology,LiveCyte live-cell tracking system,and Western blotting.Results·Analysis of the GEPIA2 database indicated that FADD was significantly overexpressed in head and neck cancer and was associated with poor prognosis.IHC staining showed that FADD expression levels progressively increased from normal to dysplastic to tumor tissues in HNSCC patients.Knockdown of FADD in HNSCC cells resulted in significantly reduced proliferation and colony formation compared to the control group.Co-IP/MS results showed that FADD interacted with the CUX1 protein,and FADD knockdown led to increased CUX1 expression.Moreover,CUX1 knockdown significantly promoted HNSCC cell proliferation and reversed the anti-proliferative phenotype caused by FADD knockdown.Conclusion·FADD plays a significant pro-carcinogenic role in HNSCC and is associated with poor prognosis.FADD can further regulate tumor cell proliferation by interacting with CUX1 and suppressing its expression level.
6.Research progress of finite element modeling for clear aligner treatment
Yiyan ZHANG ; Cunhui FAN ; Hanlin DING ; Qianwen ZHANG ; Yang LIU
Chinese Journal of Stomatology 2025;60(7):800-806
Finite element method, as a verified numerical simulation method, has been widely applied in the biomechanical research of clear aligner treatment. The accuracy of its calculation results is closely related to the construction method of the finite element model. This paper takes the finite element modeling process of clear aligner treatment as a clue, and reviews the modeling methods of relevant research in recent years, aiming to provide reference for subsequent studies.
7.Progress of serious games in motor function rehabilitation of patients with skeletal muscle injuries
Huijuan QIAN ; Qunfeng LU ; Huiling YUE ; Hanlin YANG ; Changrong YUAN
Chinese Journal of Modern Nursing 2025;31(19):2535-2539
By measuring how patients with skeletal muscle injuries move and assisting healthcare professionals in instructing patients to perform specific movements in a gamified manner, serious games can effectively increase exercise adherence and positively improve clinical outcomes for patients, ultimately resulting in reduced health education costs and accessible rehabilitation programs. This paper describes the conceptual framework for the creation of serious games and their content characteristics, reviews the training methods and effects of serious games in the motor function rehabilitation among patients with skeletal muscle injuries, and puts forward the future prospects for the application of serious games in motor function rehabilitation.
8.Trajectories of executive function development and its neural mechanisms in patients with attention deficit hyperactivity disorder
Ruilin JIN ; Jiaqi ZHOU ; Teng ZHU ; Jiayun YU ; Wanying ZHENG ; Hanlin LI ; Mengjie ZHANG ; Xiaolei CEN ; Chuang YANG
Chinese Journal of Behavioral Medicine and Brain Science 2025;34(3):277-282
Executive function(EF) is an advanced cognitive function of the central nervous system, and is closely related to an individual's capacity for daily living and adaptation. Patients with attention deficit hyperactivity disorder (ADHD) typically exhibit significant executive dysfunction. While most existing studies on the executive function of individuals with ADHD are cross-sectional, and little is known about the longitudinal maturation process of related brain structures and functional connectivity patterns. The findings indicate that ADHD patients exhibit differential developmental trajectories in brain structural and functional connectivity compared with typically developing group.Furthermore, there is a lifespan association between abnormal brain network development and ADHD symptoms. This article aims to elucidate the characteristics of executive function deficits in ADHD patients across different developmental stages, examining their relationship with the nervous system’s development from a development perspective.
9.Research progress of finite element modeling for clear aligner treatment
Yiyan ZHANG ; Cunhui FAN ; Hanlin DING ; Qianwen ZHANG ; Yang LIU
Chinese Journal of Stomatology 2025;60(7):800-806
Finite element method, as a verified numerical simulation method, has been widely applied in the biomechanical research of clear aligner treatment. The accuracy of its calculation results is closely related to the construction method of the finite element model. This paper takes the finite element modeling process of clear aligner treatment as a clue, and reviews the modeling methods of relevant research in recent years, aiming to provide reference for subsequent studies.
10.A study of the effectiveness of an ICU transitional care program in improving relocation stress and caregiving skills of parents of critically ill children
Hanlin YANG ; Jianlin JI ; Chengxi ZENG ; Ouyao CHEN ; Qunfeng LU
Chinese Journal of Nursing 2025;60(16):1966-1973
Objective Constructing a transitional care program for critically ill children in the ICU and exploring the effects of its application on the relocation stress and caregiving abilities of parents of children to improve their caregiving experience.Methods Convenience sampling method was used to select 150 children and their families transferred from PICU to the general ward of a tertiary pediatric hospital in Shanghai from January to September 2023 as research subjects.Subjects enrolled from June to September 2023 were selected as an experimental group,and were applied with the ICU transitional care program for critically ill children.Those enrolled from January to April 2023 were selected as a control group,and received routine ICU transitional care.Family Relocation Stress Scale and Family Caregiver Task Inventory were used to evaluate children's families at 24h before,24h after,and 72h after children's transition from ICU.The incidence of unplanned readmission to ICU within 48h and adverse events were compared between 2 groups of children.Results Eventually 74 pairs in the experimental group and 75 pairs in the control group completed the study.The Family Caregiver Task Inventory scores in the experimental group were lower than that in the control group(P=0.046).Generalized estimation equation showed that,there were significant differences on the intervention effect,the time effect,and the interaction effect of the Family Relocation Stress Scale scores in 2 groups(P<0.05).Conclusion The ICU transitional care program for critically ill children constructed in this study can effectively improve the level of migration stress and caregiving ability of parents of critically ill children,and optimize the quality of transitional care for critically ill children.

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