1.In vitro cultured calculus bovis alleviates cerebral ischemia/reperfusion injury through regulating microglial polarization and inhibiting NLRP3.
Tanlu CHU ; Wei ZHANG ; Jingwen CHEN ; Zeyue PAN ; Lingfeng WANG ; Xiaoming ZHONG ; Fengmei QIU ; Zhen HUANG
Journal of Zhejiang University. Medical sciences 2025;54(3):360-371
OBJECTIVES:
To investigate the effect of in vitro cultured calculus bovis (ICCB) on cerebral ischemia/reperfusion injury (CIRI) and its mechanism.
METHODS:
A CIRI rat model and a cell model were induced by middle cerebral artery occlusion (MCAO) in Sprague Dawley rats and oxygen glucose deprivation/reperfusion (OGD/R) in BV2 cells, respectively. The CIRI rat model was evaluated using the modified neurological severity score (mNSS), brain water content, and cerebral infarction volume after 1.5 h of ischemia followed by 72 h of reperfusion. Histopathological changes in the cortex and hippocampal CA1 region were observed with hematoxylin and eosin staining. Microglial polarization and NOD-like receptor thermal protein domain associated protein (NLRP) 3 inflammasome expression in the cortex were examined by immunofluorescence. BV2 cell viability was measured via MTT assay after treatment with ICCB and Nigericin. The expressions of NLRP3, ASC, caspase-1 proteins and inflammatory cytokines were detected with Western blotting in OGD/R treated BV2 cells (0.5 h OGD+24 h reperfusion) and in cells pretreated with Nigericin for 24 h.
RESULTS:
ICCB treatment significantly improved neurological function, reduced cerebral infarct volume and brain water content, and mitigated pathological damage in the cortical and hippocampal CA1 regions of rats subjected to CIRI (all P<0.05). ICCB promoted the transition of cortical microglia from M1 to M2 phenotypes and suppressed NLRP3 activation in microglial cells (all P<0.01). ICCB significantly down-regulated the expression of NLRP3, ASC, and caspase-1 proteins, and reduced the secretion of IL-18 and IL-1β in BV2 cells of OGD/R model (all P<0.01). In addition, Nigericin significantly reversed the salvage effect of ICCB on model cells (both P<0.01) and the modulation of inflammatory cytokines (P<0.05).
CONCLUSIONS
ICCB exerts a protective effect against CIRI by mitigating neuroinflammation, through the reduction of M1 microglial polarization, promotion of M2 conversion, and suppression of the NLRP3/ASC/caspase-1 signaling pathway.
Animals
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Rats, Sprague-Dawley
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Reperfusion Injury/prevention & control*
;
Microglia/metabolism*
;
Rats
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NLR Family, Pyrin Domain-Containing 3 Protein
;
Brain Ischemia/metabolism*
;
Male
2.Simulation Study of Myocardial Tissue Ablation Effects Using Flower Petal-Structured Electrodes in Pulsed Ablation.
Chinese Journal of Medical Instrumentation 2025;49(2):125-133
This study aims to evaluate the application of flower petal-structured electrodes in pulsed field ablation (PFA) technology, with a particular focus on their performance in myocardial tissue ablation. Through a combination of simulation techniques and in vitro experiments, the study investigates the effects of different voltage levels, electrode-to-tissue contact distances, and their impact on ablation depth, continuity, and transmurality. The research methods include the construction of a myocardial tissue simulation model, electric field distribution simulation using COMSOL Multiphysics, and in vitro ablation experiments on potato tissue. The results indicate that as voltage increases, the ablation depth significantly increases. At a voltage of 2500 V, a transmural ablation depth of 4 mm can be achieved, and the ablation area remains relatively continuous. The in vitro experiments confirm the consistency of the simulation results, and pulsed field ablation does not induce significant temperature rise, confirming its non-thermal characteristic. The conclusion suggests that PFA technology requires less electrode contact and offers higher ablation efficiency, providing a new technological pathway for the clinical treatment of atrial fibrillation and effectively reducing the risk of complications associated with traditional ablation techniques.
Electrodes
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Catheter Ablation/instrumentation*
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Computer Simulation
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Flowers
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Atrial Fibrillation/surgery*
;
Myocardium
4.Association of NLRP3 genetic variant rs10754555 with early-onset coronary artery disease.
Lingfeng ZHA ; Chengqi XU ; Mengqi WANG ; Shaofang NIE ; Miao YU ; Jiangtao DONG ; Qianwen CHEN ; Tian XIE ; Meilin LIU ; Fen YANG ; Zhengfeng ZHU ; Xin TU ; Qing K WANG ; Zhilei SHAN ; Xiang CHENG
Chinese Medical Journal 2025;138(21):2844-2846
5.Evaluation of effectiveness of online courses on sexuality education for rural girls in China
GUO Lingfeng, LIU Shuang, CHEN Jialing, LI Xuelu
Chinese Journal of School Health 2024;45(9):1285-1289
Objective:
To explore the impact of online sexuality education courses on the sexual knowledge, attitudes and related behaviors of rural girls, so as to provide the practical guidance for promoting sexual health and development.
Methods:
From February to June 2023, by posting information online and through commonweal organization websites, rural primary schools in 12 provinces were recruited for a semester of online sexuality education courses from October to November 2023. A self compiled sexuality education questionnaire was used to survey the knowledge, attitudes, and behaviors of rural girls before and after the course intervention, with pre tests in September 2023 and post tests in December 2023. The eligible samples were 3 058 and 2 602 , respectively. Independent sample t tests and text frequency and sentiment analysis were used to process the data
Results:
In terms of sexual knowledge, the scores of rural girls before and after the test were (8.49±3.29) and (9.40±3.35), respectively, with the post test score being higher than the pre test ( t =-10.20, P <0.01). In terms of attitudes, the scores of rural girls before and after the test were (11.50±4.62) and (10.82±4.80), respectively, with a decrease in stigmatization towards physiological development in the post test ( t =5.40, P <0.01). Regarding sexual related behaviors, the frequency of sexualbased bullying among rural girls was (5.12±2.13) before and (4.89±2.18) after, with a statistically significant difference ( t =3.99, P <0.01). The frequency and willingness of rural girls to discuss sexual topics with significant others both increased on post test (pre test:8.45±2.62; post test: 8.73± 2.62) and (pre test:8.90±2.46, post test:9.16±2.46), with a statistically significant difference ( t =-3.91, -4.03, P < 0.01 ). Text analysis revealed that "boys" "girls" and "menstruation" were the most concerned topics among the participants, and compared to before receiving sex education (69.91%), the proportion of negative emotions among rural girls decreased ( 18.59 %).
Conclusion
Online sexuality education courses can improve the sexual knowledge of rural girls, reduce stigma and negative emotions towards sex, decrease the incidence of sexual based bullying and increase the frequency and willingness to discuss sexual topics with parents, teachers, and peers.
6.Quantitative evaluation of long-term care policies for older people in the 14th Five-Year Plan in China:based on Policy Modeling Consistency index model
Lifang ZHOU ; Zihan LANG ; Yixuan WU ; Na XU ; Lingfeng XU ; Zhongming CHEN ; Wenqiang YIN ; Qianqian YU
Chinese Journal of Rehabilitation Theory and Practice 2024;30(8):939-947
Objective To quantitatively evaluate the structure and content of the long-term care policy for the older people in the 14th Five-Year Plan in China. Methods The primary indicators were constructed based on multi-source flow model,using Policy Modeling Consisten-cy(PMC)model,and the secondary indicators were constructed based on World Health Organization reports Long-term care for older people:Package for universal health coverage and Decade of healthy ageing:Plan of action.An evaluation system was formed,containing nine primary indicators and 35 secondary indicators.A total of eight policies issued by the nation between November,2021 and January,2024 were included.High-frequency words were extracted using ROSTCM 6.0,a semantic network map of keywords was created using Gephi,and the policies were analyzed using the evaluation system. Results The policies mainly focused on nursing,community,rehabilitation and nursing home,etc.The average PMC in-dex was 6.77.For the policy content coverage,one policy was perfect,five were excellent and two were accept-able.In the dimension of policy content,the indexes of three primary indicators of policy function,policy recep-tors and infrastructure construction were 0.85 or more;while the indexes of three primary indicators of caregiver assistance,support for the elderly and human resources building were 0.7 or less. Conclusion The content of long-term care policies for the older people issued in the 14th Five-Year Plan in China covers well in policy functions,policy recipients and infrastructure,and needs to be upgraded in the three dimensions of caregiver assistance,support for the elderly and human resource building.It is important to strengthen the policy content and measures in three areas,namely,the comprehensive care support network,the creation of an age-friendly environment and support for informal care.
7.Research on the application of monitoring and regulation of intrapelvic pressure in supermicropercutaneous nephrolithotomy
Bin CHEN ; Lingfeng WU ; Linfeng LU ; Yifang CAO ; Wei CHEN ; Xueping WANG ; Wenhua XIE
China Modern Doctor 2024;62(14):60-64
Objective To investigate the application of intra pelvic pressure(IPP)in ultramicro-channel percutaneous nephrolithotripsy.Methods From January 2022 to January 2023,60 patients with urinary calculi who needed Super mini-PCNL(SMP)in The First Hospital of Jiaxing selected as the study objects.According to random number method,the patients were divided into control group and experimental group,with 30 cases.Both groups were treated with ultra-micro channel percutaneous nephrolithotripsia,while the experimental group was monitored and regulated IPP in real time during the operation,and observed and compared clinical indicators,IPP,fever,urinary protein,renal function,hemoglobin(Hb)and adverse reactions between the two groups.Results Compared with the control group,the hospitalization time of experimental group was shortened and the stone clearance rate was increased(P<0.05).The IPP levels of experimental groups at 6min,12min,24min and 36min were lower than those of control group(P<0.05).The fever of experimental group was lower than that of control group at 2d,3d,4d and 5d after operation(P<0.05).The urinary protein level of experimental group was lower than that of control group at 1d,2d,3d and 4d after operation(P<0.05).Compared with control group,blood urea nitrogen(BUN)and serum creatinine(SCr)levels of experimental group were decreased,and Hb levels were increased(P<0.05).The incidence of adverse reactions in experimental group was lower than that in control group(P<0.05).Conclusion Monitoring and adjusting intrapelvic pressure during super-mini percutaneous nephrolithotomy is beneficial in reducing postoperative fever in patients with urolithiasis,reducing urinary protein expression and kidney function damage,and controlling the occurrence of adverse reactions.It is worth recommending.
8.Costunolide covalently targets NACHT domain of NLRP3 to inhibit inflammasome activation and alleviate NLRP3-driven inflammatory diseases.
Haowen XU ; Jiahao CHEN ; Pan CHEN ; Weifeng LI ; Jingjing SHAO ; Shanshan HONG ; Yi WANG ; Lingfeng CHEN ; Wu LUO ; Guang LIANG
Acta Pharmaceutica Sinica B 2023;13(2):678-693
The NLRP3 inflammasome's core and most specific protein, NLRP3, has a variety of functions in inflammation-driven diseases. Costunolide (COS) is the major active ingredient of the traditional Chinese medicinal herb Saussurea lappa and has anti-inflammatory activity, but the principal mechanism and molecular target of COS remain unclear. Here, we show that COS covalently binds to cysteine 598 in NACHT domain of NLRP3, altering the ATPase activity and assembly of NLRP3 inflammasome. We declare COS's great anti-inflammasome efficacy in macrophages and disease models of gouty arthritis and ulcerative colitis via inhibiting NLRP3 inflammasome activation. We also reveal that the α-methylene-γ-butyrolactone motif in sesquiterpene lactone is the certain active group in inhibiting NLRP3 activation. Taken together, NLRP3 is identified as a direct target of COS for its anti-inflammasome activity. COS, especially the α-methylene-γ-butyrolactone motif in COS structure, might be used to design and produce novel NLRP3 inhibitors as a lead compound.
9.Investigation on the mechanism of Compound zaoren granules in improving insomnia based on serum meta-bonomics
Zekun WANG ; Shenlin LIU ; Xiaocong YU ; Danting LI ; Lingfeng ZHANG ; Yimeng ZHAO ; Chen CHEN ; Yajun CHEN ; Yachun SHU
China Pharmacy 2023;34(9):1093-1098
OBJECTIVE To study the mechanism of Compound zaoren granule in improving insomnia. METHODS Forty-nine mice were divided into blank group, model group, positive control group 1 (Estazolam tablets 0.5 mg/kg),control group 2 (Shumian capsule 0.6 g/kg), Compound zaoren granule low-dose, medium-dose and high-dose groups (2.5, 5, 10 g/kg), with 7 mice in each group. The insomnia model was established by chronic unpredictable mild stress combined with 4-chloro-DL- phenylacetic acid. The behavioral changes of mice were investigated through open field test and pentobarbital sodium synergistic hypnosis experiment, as well as the pathomorphology of mice hypothalamus tissue was observed by HE staining. The metabonomics analysis and multivariate statistical analysis of serum in mice were performed by UHPLC-Q-TOF-MS/MS, and the differential metabolites were screened out; the metabolic pathway analysis was conducted based on MetaboAnalyst 5.0 database. RESULTS Compared with blank group, the total travelling distance, the number of entering the central region and the moving distance in the central region of the model group were significantly reduced (P<0.05), the proportion of total rest time was significantly increased (P<0.05), the sleep duration of mice was significantly shortened (P<0.05), and hypothalamic nerve cells damaged and severely vacuolated. Compared with model group, the total travelling distance of Compound zaoren granule low-dose and medium-dose groups were increased significantly and the proportions of total rest time of those groups were decreased significantly (P<0.05), and the sleep duration of mice in Compound zaoren granule high-dose group was prolonged significantly (P<0.05); the hypothalamic nerve cells of mice in each administration group recovered to varying degrees, and the hypothalamus histiocytes of mice in the Compound zaoren granules high-dose group were closer to those in the blank group. A total of 18 differential metabolites (such as phenylalanine, taurine, norvaline, methionine) and 4 important amino acid metabolic pathways (L-phenylalanine, tyrosine and tryptophan biosynthesis; taurine and hypotaurine metabolism; L-phenylalanine metabolism; cysteine and methionine metabolism) were identified through metabolomics analysis. CONCLUSIONS Compound zaoren granules can normalize the disordered metabolism in vivo by regulating differential metabolites such as phenylalanine, taurine, and four amino acid metabolic pathways, so as to improve insomnia.


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