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.Zuoguiwan Mitigates Oxidative Stress in Rat Model of Hyperthyroidism Due to Kidney-Yin Deficiency via DRD4/NOX4 Pathway
Ling LIN ; Qianming LIANG ; Changsheng DENG ; Li RU ; Zhiyong XU ; Chao LI ; Mingshun SHEN ; Yueming YUAN ; Muzi LI ; Lei YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):43-51
ObjectiveTo decipher the mechanism by which Zuoguiwan (ZGW) treat hyperthyroidism in rats with kidney-Yin deficiency based on the dopamine receptor D4 (DRD4)/nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 4 (NOX4) signaling pathway. MethodsThe rat model of kidney-Yin deficiency was induced by unilateral intramuscular injection of dexamethasone (0.35 mg·kg-1). After successful modeling, the rats were randomized into model, methimazole (positive control, 5 mg·kg-1), low-, medium-, and high-dose (1.85, 3.70, 7.40 g·kg-1, respectively) ZGW, and normal control groups. After 21 days of continuous gavage, the behavioral indexes and body weight changes of rats were evaluated. The pathological changes of the renal tissue were observed by hematoxylin-eosin staining. The serum levels of thyroid hormones [triiodothyronine (T3), thyroxine (T4), thyroid-stimulating hormone (TSH)], renal function indexes [serum creatine (Scr) and blood urea nitrogen (BUN)], energy metabolism markers [cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP)], and oxidative stress-related factors [superoxide dismutase (SOD), malondialdehyde (MDA), and NADPH)] were measured by enzyme-linked immunosorbent assay (ELISA). Western blot was employed to analyze the expression of DRD4, NOX4, mitochondrial respiratory chain complex proteins [NADH:ubiquinone oxidoreductase subunit S4 (NDUFS4) and cytochrome C oxidase subunit 4 (COX4)], and inflammation-related protein [tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), p38 mitogen-activated protein kinase (MAPK)] pathway in the renal tissue. ResultsCompared with the normal group, the model group showed mental malaise, body weight decreases (P<0.01), inflammatory cell infiltration in the renal tissue, a few residual parotid glands in the thyroid, elevations in serum levels of T3, T4, Scr, BUN, cAMP, cAMP/cGMP, MDA, and NADPH (P<0.01), down-regulation in protein levels of TSH, SOD, and DRD4 (P<0.05, P<0.01), and up-regulation in expression of NOX4, p-p38 MAPK/p38 MAPK, and inflammatory factors (P<0.01). Compared with the model group, ZGW increased the body weight (P<0.05, P<0.01), reduced the infiltration of renal interstitial inflammatory cells, restored the thyroid structure and follicle size, lowered the serum levels of T3, T4, Scr, BUN, cAMP, cAMP/cGMP, MDA and NADPH (P<0.05, P<0.01), up-regulated the expression of TSH, SOD and DRD4 (P<0.05, P<0.01), and down-regulated the expression of NOX4, p-p38 MAPK/p38 MAPK, and inflammatory factors (P<0.05, P<0.01). Moreover, high-dose ZGW outperformed methimazole (P<0.05). ConclusionBy activating DRD4, ZGW can inhibit the expression of NOX4 mediated by the p38 MAPK pathway, reduce oxidative stress and inflammatory response, thereby ameliorating the pathological state of hyperthyroidism due to kidney-Yin deficiency. This study provides new molecular mechanism support for the clinical application of ZGW.
3.Syndrome Patterns Distribution and Risk Factors of Mixed Hemorrhoids in Traditional Chinese Medicine: A Multicenter Real-world Study Using Large Language Models and Latent Class Analysis
Ruyue DENG ; Kang DING ; Yuxin ZHU ; Meng LI ; Huiting ZHU ; Lei DU
Journal of Traditional Chinese Medicine 2026;67(7):755-763
ObjectiveTo develop a standardized classification model for traditional Chinese medicine (TCM) syndrome patterns of mixed hemorrhoids using multi-center real-world data, and unveil their distribution patterns and core risk factors, thereby providing evidence-based support for standardizing TCM syndrome differentiation and implementing precision interventions. MethodsA multi-center cross-sectional study was conducted, enrolling 13 283 mixed hemorrhoid patients from eight hospitals in Jiangsu Province between September 1st, 2023 and December 31st, 2024. DeepSeek-R1-Distill-Qwen-7B and LLaMA-3.3 large language models (LLM) were integrated with latent class analysis (LCA) to perform unsupervised learning and latent class modeling of TCM symptomatology. Potential risk factors were screened via univariate analysis, followed by logistic regression to identify independent risk factors for each syndrome pattern. ResultsThe model's performance indicators were stable and reliable across different clinical data types,i.e. in the outpatient records, past medical history (F1=99.7%), current medical history (F1=94.9%), and specialist examination (F1=90.7%); in inpatient records, past medical history (F1=98.2%), current medical history (F1=91.2%), specialist examination (F1=90.3%), and discharge status (F1=90.6%). Latent class mode-ling identified four core TCM syndrome patterns including spleen deficiency and qi sinking syndrome (915 cases, 6.89%), damp-heat pouring downward syndrome (10 820 cases, 81.46%), qi stagnation and blood stasis syndrome (1252 cases, 9.43%), and wind injuring intestinal collaterals syndrome (296 cases, 2.22%), with respective latent class probabilities of 0.069, 0.815, 0.094, and 0.022. Logistic regression demonstrated that gender, age, disease duration, hypertension, diabetes, hyperlipidemia, constipation, smoking history, and alcohol consumption were independent risk factors for pattern differentiation (P<0.05). The efficacy validation evaluation revealed that the cure rates for patients with spleen deficiency and qi sinking syndrome and qi stagnation and blood stasis syndrome were higher than those for patients with damp-heat pouring downward syndrome (adjusted P<0.05), with no statistically significant differences among other syndrome patterns. ConclusionDamp-heat pouring downward syndrome is the predominant syndrome in mixed hemorrhoids. Gender, age, disease duration, hypertension, diabetes, hyperlipi-demia, constipation, smoking history, and alcohol consumption are independent risk factors for the differentiation of syndrome types.
4.Evaluation of repeated testing with blood screening platform in confirmation of NAT non-discriminatory reactive samples
Mengfan LI ; Xuelian DENG ; Liang ZANG ; Lei ZHOU ; Xiaochun LIU ; Xiaohua LIANG ; Lunan WANG
Chinese Journal of Blood Transfusion 2026;39(4):458-464
Objective: To evaluate repeated testing on blood screening platforms in confirmation of non-discriminatory reactive (NDR) samples in nucleic acid testing (NAT). Methods: A total of 102 HBsAg-negative/NAT NDR samples were collected from voluntary blood donors at Dalian Blood Center between January 2021 and December 2023. Repeated testing was performed using two NAT platforms (Cobas s201 and Panther). For the first round of repeated testing, all samples were tested 12 times on each system; for the second round, the samples which were non-reactive or only reactive once in the first round were tested an additional 8 times. Anti-HBc and anti-HBs was detected using electrochemiluminescence assay (ECA). Meanwhile, blood donors were followed up. Results: The proportion of anti-HBc+ in 102 NDR samples was 88.2%. Forty-one samples (40.2%, 41/102) and 7 samples were confirmed HBV DNA+ in first-round and second-round repeated testing, respectively. The cumulative confirmation rate of HBV DNA+ was 47.1% (48/102) after repeated testing. Extra five blood donors detected HBV DNA+ in follow-up were identified as anti-HBc+ occult hepatitis B virus infection (OBI), while no window period infection was observed. Ultimately, there were 53 HBV infected donors confirmed, 46 HBV infection-unconfirmed, and 3 HBV uninfected. No significant difference was observed between the confirmation rate of the first-round testing and the cumulative confirmation rate after the second-round testing (P>0.05). The proportion of anti-HBc+ donors was quite high in both HBV infection-confirmed (98.1%) and unconfirmed group (82.6%), and donors with seronegative and anti-HBs-only occupied a high proportion in the latter (P<0.05). Conclusion: Numerous repeated testing of NDR samples using NAT platforms cannot achieve complete confirmation of HBV infection. Supplementary anti-HBc testing can minimize potential OBI risk among NDR donors, and is low-cost and efficient.
5.Establishment and assessment of deep vein thrombosis model in rats in a plateau hypoxic environment
Xiaobo HAN ; Yahao CHAI ; Jiawei GAO ; Xinkai DENG ; Xiao LI ; Jialin WU ; Xiaoli HAN ; Guoxiang LI ; Yinjie ZHAO ; Xi YANG ; Qi AO ; Lei ZHANG ; Hongwei HAN ; Zhixue LIU
Acta Laboratorium Animalis Scientia Sinica 2025;33(8):1133-1143
Objective To establish a rat model of venous thrombosis in a plateau hypobaric hypoxic environment and to investigate the effect of this environment on venous thrombosis.Methods A total of 144 healthy male SD rats were assigned randomly to four groups(n=36 rats per group):a plains sham operation(A)group,plains operation(B)group,plateau altitude 6000 m+sham operation(C)group,and plateau altitude 6000 m+surgery(D)group.Rats in A and B groups were maintained in a plains normoxic environment,while rats in C and D groups C and D were subjected to a plateau environment.Rats in the surgical groups underwent quantitative constriction to incompletely obstruct the inferior vena cava blood flow.Each group was further divided into subgroups based on time:1,3,5,7,14,and 21 d(n=6 rats per group).Regular vascular ultrasound monitoring was conducted,and blood samples were taken for whole blood viscosity testing and the assessment of inflammatory indicators,including endothelin-1(ET-1),interleukin-6(IL-6)and tissue factor(TF).Coagulation function was evaluated through the activated partial thromboplastin time(APTT),prothrombin time(PT),thrombin time(TT),fibrinogen(FIB)and D-dimer.After the observation period,the experimental animals were sacrificed and the limbs were removed.Thrombus samples were stained with hematoxylin/eosin(HE),and the thrombus wet mass was measured.Results The thrombosis incidence was significantly higher in the plateau D group than in B group,accompanied by a marked increase in blood viscosity and hematocrit(P<0.01).Additionally,levels of ET-1,IL-6,and TF were significantly elevated(P<0.05),indicating a coagulation disorder.Conclusions A plateau hypoxic environment model can be successfully simulated by quantitative coarctation of the inferior vena cava,combined with a specialized environmental chamber.The findings of this study suggest that a plateau hypoxic environment promotes venous thrombosis.
6.Treatment and management of patients with endemic fluorosis in Ankang City, Shaanxi Province from 2019 to 2023
Ying DENG ; Tonglei ZHANG ; Lei LIANG ; Feng SU ; Xiaoqian LI ; Rong ZHOU ; Zhongxue FAN ; Lu WANG
Chinese Journal of Endemiology 2025;44(2):137-141
Objective:To investigate the treatment and management of patients with endemic fluorosis (referred to as fluorosis) in Ankang City, Shaanxi Province, and to provide a scientific basis for adjusting and optimizing treatment and management strategies for fluorosis patients.Methods:A retrospective analysis was conducted on patient information collected from county-level disease prevention and control centers in Ankang City from 2019 to 2023. Data included demographic information, clinical treatment, chronic disease management, treatment records, work reports, and summaries. The study aimed to learn about the treatment willingness, methods, and effects of patients with fluorosis, and to evaluate the disease management situation of patients with fluorosis.Results:From 2019 to 2023, the proportion of fluorosis patients who were willing to receive treatment in Ankang City had increased from 76.80% (1 089/1 418) in 2019 to 100% (1 408/1 408) in 2023, showing an upward trend year by year (χ 2trend = 971.15, P < 0.001). A total of 6 876 patients received treatment, with a treatment rate of 100%. The majority of patients received drug treatment (99.64%, 6 851/6 876), with an effective rate of 94.90% (6 525/6 876). The effective rate of treatment for patients with fluorosis had increased from 88.71% (966/1 418) in 2019 to 94.67% (1 333/1 408) in 2023, showing an upward trend (χ 2trend = 1 002.92, P < 0.001). The adverse reaction rate of drug treatment was 0.36% (25/6 876). The combination of oral medication and local treatment had the highest effective rate (97.62%, 3 566/3 653). The management rate and contracted service rate of fluorosis patients were 100% each year, and the standardized management rate was above 99%. Conclusions:Fluorosis patients in Ankang City have a high willingness to receive treatment, with good treatment outcomes. The contracted services and management of patients are well implemented. In the future, continuous efforts should be made to improve the prevention, management, and health education of fluorosis patients.
7.Mechanism of action of Sterculiae Lychnophorae Semen against PM2.5-induced acute lung injury based on network pharmacology,molecular docking and experimental validation
Fan ZHANG ; Yi-fan DU ; Xiao-shu DENG ; Zu-feng ZHANG ; Xian-lei HAN ; Wei TIAN ; Xiu-mei LI ; Mian CHEN ; Fei LIU ; Nan WANG
Chinese Pharmacological Bulletin 2025;41(12):2362-2369
Aim To investigate the anti-acute lung injury(ALI)effect of Sterculiae Lychnophorae Semen(SLS)and its mechanism.Methods The main ac-tive components of SLS and their core targets and path-ways of action against ALI were obtained by network pharmacology methods.Subsequently,molecular doc-king technology and in vitro cellular experiments were applied for validation.Results A total of 19 core tar-gets were obtained,including HSP90AA1,CASP3,TNF,MAPK8 and MAPK14.The mechanisms may in-volve signaling pathways such as cancer,PI3K/Akt and MAPK.Molecular docking confirmed that the key targets of SLS formed a better binding activity with the relevant active ingredients.The in vitro results showed that SLS was able to protect the PM2.5-contaminated BEAS-2B cells,inhibit their NO,IL-1β and TNF-αlevels,and reduce the expression of p-p38 MAPK and p-JNK proteins.Conclusions The study successfully predicts the active ingredients,targets and signaling pathways of SLS against ALI,and in vitro experiments demonstrate that SLS might protect BEAS-2B cells from PM2.5 stimulus-induced inflammation and apoptosis by inhibiting the over-activation of p38 MAPK and JNK signaling pathways.
8.Treatment of aneurysmal bone cyst in children by percutaneous injection of calcitonin and methylprednase
Yong LI ; Zhu WEN ; Wulan MAI ; Jun LEI ; Lin CAI ; Zhouming DENG
Journal of Clinical Surgery 2025;33(7):750-753
Objective To investigate the clinical effect of percutaneous injection of calcitonin and methylprednone in the treatment of aneurysmal bone cyst in children.Methods From June 2020 to March 2023,19 patients with aneurysmal bone cysts were admitted and all were treated with percutaneous puncture injection of calcitonin and methylprednisolone.After percutaneous puncture biopsy,200 iu of calcitonin and 120 mg of methylprednone were injected into the cyst of aneurysmal bone cyst.Compare the volume of the cyst cavity and the thinnest cortical bone thickness around the cyst cavity before and after the operation.All patients were followed up,while preoperative and postoperative tumor cavity and the thinnest peritumor cortical thickness were compared.Results 19 patients were followed up for an average of 16 months(12-52 months).Of the 19 patients,17 were effective,the effective rate was 89.5%,and no serious complications occurred in all cases.At the last follow-up,the volume of the tumor cavity was significantly lower than that before surgery[(37.05±21.17)cm3,(110.95±45.67)cm3],and the difference was statistically significant(P<0.05).The average thinnest cortical bone thickness around the tumor cavity was(0.71±0.37)mm before surgery and(1.87±0.60)mm after surgery.The difference was statistically significant(P<0.05).Conclusion Percutaneous injection of calcitonin and methylprednone in the treatment of aneurysmal bone cysts in children is minimally invasive,safe and effective.It can be used as the first-line treatment for aneurysmal bone cyst in children.
9.Analysis of 131I internal exposure levels of nuclear medicine staff in Jiangxi province from 2022 to 2023
Zhe WANG ; Lei DENG ; Faming CAO ; Li TAN
Chinese Journal of Radiological Medicine and Protection 2025;45(6):526-530
Objective:To understand the current status of thyroid 131I internal exposure of nuclear medicine staff in Jiangxi province and analyze its influencing factors. Methods:An survey was conducted for the years 2022 to 2023, involving 211 nuclear medicine staff who had received 131I treatment in Jiangxi province. The 131I activity in thyroid was measured by in vitro monitoring measurement, and the committed effective dose was estimated. Results:In 2022, 14 nuclear medicine staff were detected to have 131I in thyroid, with activities ranging from 121.32 to 2 859.09 Bq, including four staff who were estimated to have received committed effective doses above 2 mSv. In 2023, there were 21 nuclear medicine staff who were detected to have 131I in thyroid, with activities ranging from 81.75 to 1 482.21 Bq, in which 10 staff were estimated to have the committed effective dose above 2 mSv. There were no statistically significant differences between the two years in detection rate, measured activity, and committed effective dose to thyroid from 131I ( P>0.05). The highest valure of detection rate was found in cleaning staff, and there were no statistically significant differences between different position types ( P>0.05). There was no statistically significant difference in the detection rate of 131I between those who only have received hyperthyroidism treatment and those who have received both hyperthyroidism and thyroid cancer treatment ( P>0.05). Conclusions:The issue of internal exposure of nuclear medicine staff should receive attention. It is necessary to further carry out internal exposure monitoring. Meanwhile, it is suggested that the management of radiation protection should be strengthened in nuclear medicine workplaces on the part of hospitals.
10.Association Between Resting Heart Rate and Atherosclerotic Cardiovascular Disease and All-cause Death in Young and Middle-aged Adults
Lei LI ; Qi QI ; Xinyu WU ; Quanle HAN ; Jie DENG ; Xuechao ZHANG ; Zheng WU ; Nan WANG ; Shouling WU ; Kangbo LI
Chinese Circulation Journal 2025;40(7):681-688
Objectives:To investigate the correlation between resting heart rate(RHR)and atherosclerotic cardiovascular disease(ASCVD)and all-cause death in young and middle-aged people.Methods:A prospective cohort study was conducted enrolling 72 642 young and middle-aged participants(aged<60 years),who participated in the medical examination of the Kailuan Study from June 2006 to October 2007.According to the quartile of the RHR level,the participants were divided into Q1 group(<67 beats/min,n=14 381),Q2 group(67-70 beats/min,n=15 815),Q3 group(71-75 beats/min,n=15 876),Q4 group(76-80 beats/min,n=13 933)and Q5 group(>80 beats/min,n=12 637).Cox proportional hazard regression model was used to analyze the effect of RHR on ASCVD and all-cause death.The dose-response relationship between RHR and the risk of ASCVD and all-cause death was investigated using a restricted cubic spline regression model.Results:During a mean follow-up of(10.0±4.85)years,2 898 patients(3.99%)developed ASCVD.Multivariate Cox regression analysis showed that after adjusting for confounding factors,the risk of RHR and ASCVD in group Q5 increased by 20%compared with group Q1(HR=1.20,95%CI:1.06-1.35,P<0.05).There was no significant risk in groups Q2 to Q4 compared to Q1 group(all P>0.05).In addition,the risk of ASCVD increased by 4%for every 10 beats/min increase in RHR(HR=1.04,95%CI:1.01-1.07,P=0.009).During the follow-up period of(10.2±4.82)years,all-cause death occurred in 2 175 participants(2.99%).The results showed that compared with Q1 group,the risk of all-cause death in Q3 to Q5 groups increased by 33%(HR=1.33,95%CI:1.15-1.54,P<0.001),33%(HR=1.33,95%CI:1.14-1.54,P<0.001),and 78%(HR=1.78,95%CI:1.54-2.05,P<0.001)respectively,and there was no statistical significance between group Q2 and group Q1.The risk of all-cause death increased by 15%for every 10 beats/min increase in RHR(HR=1.15,95%CI:1.11-1.19,P<0.001).Restricted cubic spline analysis showed that RHR was linearly correlated with risk of ASCVD(Poverall=0.022,Pnon-linear=0.617),and the risk of ASCVD increased significantly with RHR>72 beats/min.RHR was linearly associated with the risk of all-cause death(Poverall<0.001,Pnon-linear=0.212),and the risk of all-cause death was significantly increased with RHR>72 betas/min.Conclusions:Higher RHR is associated with an increased risk of ASCVD and all-cause mortality in young and middle-aged individuals.

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