1.Mechanism of action of remifentanil in alleviating lung ischemia-reperfusion injury in rats by modulating HIF-1α/NLRP3 pathway to inhibit cell pyroptosis
Lifang ZHAO ; Jiangong YANG ; Mingyong LI ; Kun SHAO ; Changli SHEN ; Jiajie LI ; Hong ZHU ; Liangchao QU
Acta Universitatis Medicinalis Anhui 2026;61(3):395-401
ObjectiveTo investigate the mechanism of action of remifentanil (RMZL) in alleviating lung ischemia-reperfusion injury (LIRI) in rats by inhibiting pyroptosis through modulating hypoxia inducible factor-1α (HIF-1α)/NOD-like receptor thermal protein domain associated protein 3 (NLRP3) pathway. MethodsRats were stochastically assigned into Control group, LIRI group, RMZL low-dose group, RMZL medium-dose group, RMZL high-dose group, and RMZL high-dose+HIF-1α activator dimethyloxallyl glycine (DMOG) group, with 18 rats in each group. Rats in Control group only had their left pulmonary hilum free and did not undergo ischemia-reperfusion treatment. Except for the Control group, LIRI models were constructed in all other groups. Rats in LIRI group were intraperitoneally injected with an equal amount of physiological saline 15 minutes before constructing LIRI model; rats in Control group were intraperitoneally injected with an equal amount of physiological saline 15 minutes before freeing left pulmonary hilum; rats in other groups were intraperitoneally injected with corresponding dose of drug 15 minutes before constructing LIRI model. The wet/dry weight ratio of lungs was calculated. HE staining was used to study lung tissue pathology. Immunofluorescence staining was used to detect the relative fluorescence intensity of gasdermin D (GSDMD) and NLRP3 double positive cells in lung tissue. ELISA was used to detect interleukin-1β and IL-18 in lung tissue. Western blot was used to detect HIF-1α, NLRP3, cysteine-aspartic protease-1 (Cleaved caspase-1), and gasdermin D-N (GSDMD-N) proteins in lung tissue. ResultsCompared to the Control group, the LIRI group showed disordered alveolar structure, thickened alveolar septa, and abundant inflammatory cell infiltration in rats. The lung wet/dry weight ratio, relative fluorescence intensity of GSDMD and NLRP3 double positive cells in lung tissue, IL-1β, IL-18 levels, and HIF-1α, NLRP3, Cleaved caspase-1, and GSDMD-N proteins increased (P0.05). For the LIRI group, rats in the RMZL low, medium, and high-dose groups displayed attenuated alveolar septal thickening and reduced inflammatory cell infiltration. The lung wet/dry weight ratio, relative fluorescence intensity of GSDMD and NLRP3 double positive cells in lung tissue, IL-1β, IL-18 levels, and HIF-1α, NLRP3, Cleaved caspase-1, and GSDMD-N proteins declined, and the RMZL high-dose group showed the most prominent trend (P0.05). Compared with the RMZL high-dose group, rats in the RMZL high-dose+DMOG group exhibited thickened alveolar septa and more inflammatory cell infiltration, along with increased lung wet/dry weight ratio, relative fluorescence intensity of GSDMD and NLRP3 double positive cells in lung tissue, levels of IL-1β and IL-18, and protein expression of HIF-1α, NLRP3, Cleaved caspase-1, and GSDMD-N (P0.05). ConclusionRMZL may inhibit pyroptosis in LIRI rats by suppressing HIF-1α/NLRP3 pathway.
2.Improved YOLOv8 algorithm-based detection of pulmonary nodules in CT images
Chinese Medical Equipment Journal 2025;46(8):1-10
Objective To propose an improved YOLOv8 algorithm based on polarized self-attention(PSA)and deformable attention(DAT)so as to enhance the detection of pulmonary nodules in CT images.Methods A basic framework was established with a YOLOv8 model consisting of a backbone network(Backbone),a neck module(Neck)and a head module(Head).PSA was introduced into the end of the spatial pyramid pooling-fast(SPPF)of Backbone to construct a YOLOv8-PSA algorithm,and DAT was involved in the medium-scale feature layer P4 in Head to form a YOLOv8-DAT algorithm.The YOLOv8-PSA and YOLOv8-DAT algorithms were trained and validated using the CT image dataset of pulmonary nodules from public platforms,and compared with the original YOLOv8 algorithm for the detection of pulmonary nodule lesions in CT images.Results When used for pulmonary module detection of CT images,the YOLOv8-DAT algorithm had the mean average precision(mAP)in case of intersection over union threshold of 0.5(mAP50),mAP in case of intersection over union threshold of 0.5 to 0.95(mAP50-95)and precision ratio being 0.918,0.588 and 0.960 respectively,which gained advantages over the YOLOv8-PSA algorithm with mAP50,mAP50-95 and precision ratio being 0.914,0.583 and 0.945 respectively,and over the original YOLOv8 algorithm with mAP50,mAP50-95 and precision ratio being 0.911,0.564 and 0.952 respectively.Conclusion The YOLOv8-DAT algorithm detects pulmonary modules in CT images effectively,and facilitates early screening and diagnosis of pulmonary modules clinically.[Chinese Medical Equipment Journal,2025,46(8):1-10]
3.Simultaneous content determination of twelve constituents in Anshen Buxin Liuwei Pills by HPLC-MS/MS and their chemical pattern recognition
Cheng-dong LIU ; Jun LI ; Qian ZHANG ; Jing LIU ; Jing-kun LU ; Xin DONG ; Yuan-hong LIAO ; Yue-wu WANG
Chinese Traditional Patent Medicine 2025;47(9):2834-2840
AIM To establish an HPLC-MS/MS method for the simultaneous content determination of dehydrodiisoeugenol,eugenol,costiolactone,dehydrocostiolactone,quercetin,isorhamnetin,luteolin,caffeic acid,gallic acid,protocatechuic acid,ellagic acid and kaempferol in Anshen Buxin Liuwei Pills,and to make chemical pattern recognition.METHODS The analysis was performed on a 35 ℃ thermostatic Shim-pack GST-HP C18 column(2.1 mm × 100 mm,3 μm),with the mobile phase comprising of methanol-water(containing 0.1%formic acid)flowing at 0.25 mL/min in a gradient elution manner,and electron spray ionization source was adopted in positive and negative ion scanning with multiple reaction monitoring mode.Subsequently,cluster analysis,principal component analysis and orthogonal partial least square-discriminant analysis were performed.RESULTS Twelve constituents showed good linear relationships within their own ranges(r≥0.999 0),whose average recoveries were 95.38%-105.00%with the RSDs of 1.91%-5.14%.Thirteen batches of samples were clustered into 3 types,ellagic acid,dehydrocodenolactone,dehydrodiisoeugenol,protocatechuic acid,gallic acid,quercetin and kaempferol were taken as potential quality differential markers.CONCLUSION This accurate,sensitive,stable and reproducible method can be used for the quality control and evaluation of Anshen Buxin Liuwei Pills.
4.Effect of Electroacupuncture on TGF-β1/Smads Signaling Pathway and Epithelial-Mesenchymal Transition-related Protein Expression in Rats with Neurogenic Bladder After Spinal Cord Injury
Xiaojing LUO ; Chuning TIAN ; Lifen ZHAN ; Qian LI ; Roujun LIANG ; Lubo XIAO ; Yue ZHUO ; Kun AI ; Ming XU ; Hong ZHANG
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(10):120-126
Objective To observe the effects of electroacupuncture(EA)on TGF-β1/Smads signaling pathway and epithelial-mesenchymal transition(EMT)related proteins in rats with neurogenic bladder(NB)after spinal cord injury;To explore the possible mechanism of EA in improving NB fibrosis.Methods Totally 36 female SD rats were randomly selected,with 10 rats in the sham-operation group and the remaining 26 rats undergoing complete transection of the spinal cord beneath the T8 vertebrae to establish a NB rat model.The modeling rats were randomly divided into model group and EA group,with 10 rats in each group.EA group was applied to"Ciliao","Zhongji"and"Sanyinjiao",20 min per time,once a day for 7 days.The general condition of the rats in each group was observed,the ultrasound index of the bladder was detected by ultrasound technique,the bladder function of the rats was detected by urodynamics,the body mass of the rats and the wet weight of the bladder were recorded,and the bladder index was calculated.HE staining was used to observe bladder tissue morphology,the degree of bladder tissue fibrosis and bladder wall thickness were detected by Masson staining.The positive expressions of E-cadherin,N-cadherin and Vimentin in bladder tissue were detected by immunohistochemistry.The protein expressions of TGF-β1,p-Smad2/3,E-cadherin,N-cadherin and Vimentin were detected by Western blot.Results Compared with the sham-operation group,the upper and lower diameter,anteroposterior diameter,transverse diameter,bladder volume of the model group significantly increased(P<0.001),the maximum bladder pressure,leak point pressure difference,perfusion time and maximum bladder capacity significantly increased(P<0.001),the bladder index increased significantly(P<0.001),the bladder epithelial cells were thickened and arranged irregularly,the bladder collagen volume fraction and bladder wall thickness significantly increased(P<0.001),the expressions of TGF-β1,p-Smad2/3,N-cadherin and Vimentin in bladder tissue increased(P<0.01),and the expression of E-cadherin decreased(P<0.001).Compared with the model group,the upper and lower diameter,anteroposterior diameter,transverse diameter,bladder volume of the EA group decreased significantly(P<0.05),the maximum bladder pressure,leak point pressure difference,perfusion time and maximum bladder capacity significantly decreased(P<0.05),the bladder index significantly decreased(P<0.05),the thickness of the bladder epithelial cell layer became thinner and arranged more neatly,and the bladder collagen volume fraction and bladder wall thickness of the bladder were significantly reduced(P<0.05),the expressions of TGF-β1,p-Smad2/3,N-cadherin and Vimentin in bladder tissue significantly decreased(P<0.05),and the expression of E-cadherin significantly increased(P<0.05).Conclusion EA may reduce the EMT of bladder epithelial cells and decrease the degree of bladder tissue fibrosis by inhibiting the activation of the TGF-β1/Smads signaling pathway,thereby improving bladder function in NB rats after spinal cord injury.
5.Effects of Supplemented Wendan Decoction on glycolipid metabolism and PI3K/Akt/FOXO1 signalling pathway in 3T3-L1 adipocytes
Kai-yin ZHANG ; Feng-yun YAO ; Yao-yao HAN ; Jie-lin JIANG ; Lin WANG ; Wen LI ; Hong-fang YANG ; Huan-yuan ZHANG ; Yan-kun CUI
Chinese Traditional Patent Medicine 2025;47(10):3242-3248
AIM To investigate the impact of varying dosages of Supplemented Wendan Decoction on the PI3K/Akt/FOXO1 glycolipid metabolic pathway in 3T3-L1 adipocytes.METHODS The CCK-8 assay was used to determine the concentration of Supplemented Wendan Decoction-medicated serum.The mature adipocytes differentiated from 3T3-L1 preadipocytes after induction were further divided into the blank control group,the model group,the rosiglitazone group(10 mg/L),and the Supplemented Wendan Decoction groups(5%,10%,and 20%),followed by the sample collections after 48 hours of treatment.Oil red O staining quantified lipid accumulation in 3T3-L1 adipocytes;extracellular glucose levels were measured using glucose oxidase(GOD)assay;RT-qPCR analyzed mRNA expressions of IRS-1,PI3K,Akt,GLUT4,IL-6,TNF-α and IL-1β;Western blot assessed protein expressions of INSR,IRS-1,PI3K-p85,Akt,FOXO1 and GLUT4.RESULTS No significant changes in cell viability(P>0.05)were observed in 3T3-L1 preadipocytes exposed to serum containing supplemented Wendan Decoction at different concentrations for 24,48,or 72 hours.The 3T3-L1 preadipocytes held the capacity to differentiate into mature adipocytes within a 14-day induction period.Compared to the model group,all supplemented Wendan Decoction groups exhibited reduced lipid accumulation in adipocytes and downregulated mRNA expression of IRS-1,IL-6,TNF-α and IL-1β(P<0.01);the low-dose group demonstrated increased mRNA expressions of PI3K and GLUT4(P<0.05,P<0.01),alongside elevated protein expressions of INSR,IRS-1,PI3K-p85,Akt and GLUT4(P<0.05,P<0.01);the medium-dose group showed enhanced GLUT4 mRNA expression,and upregulated protein expressions of INSR and FOXO1(P<0.01).After 24 hours intervention,the high-dose Supplemented Wendan Decoction group exhibited increased glucose consumption in adipocytes(P<0.01),and elevated protein expression of INSR,Akt and FOXO1(P<0.05,P<0.01).CONCLUSION Supplemented Wendan Decoction reduces lipid accumulation in adipocytes,regulates glucose and lipid metabolism,and promotes metabolic homeostasis through PI3K/Akt/FOXO1 signaling pathway.
6.Ameliorative effects of Compound Fufangteng Mixture on cyclophosphamide-induced immunosuppression in mice
Li-na LIU ; Yu-fang SHEN ; Qin-qin WANG ; Lin-yu XIAO ; Jing-yu LIU ; Jun-ni MO ; Ren-yi-kun YUAN ; Hong-wei GAO ; Jian XIAO
Chinese Traditional Patent Medicine 2025;47(10):3249-3256
AIM To investigate the ameliorative effects of Compound Fufangteng Mixture(CFM)on cyclophosphamide(CTX)-induced immunosuppression in mice.METHODS Forty-eight male C57BL/6J mice were randomly divided into the blank control group,the model group,the levamisole hydrochloride group(40 mg/kg)and the low-dose,medium-dose and high-dose CFM groups(3.75,7.5,10 g/kg),with 8 mice in each group,and given respective intervention orally once daily for 14 days.On the 5th to 7th day of administration,with the blank control group given normal saline intraperitoneally,the other groups underwent intraperitoneal CTX injections(80 mg/kg).24 hours after the last administration,organ indices of thymus and spleen were calculated;splenic histopathological alterations were assessed by HE staining;serum levels of IL-2,IL-6 and IgG were quantified using ELISA;splenic CD4+,CD8+T lymphocytes,alongside CD86+and CD206+macrophages populations were analyzed by flow cytometry;and splenic expression of CD4,CD8 and F4/80 was evaluated by immunohistochemical staining.RESULTS In CTX-treated mice,CFM administration mitigated body weight loss;enhanced thymus weight and thymic index;ameliorated splenic immune cell populations,elevated serum levels of cytokines IL-2,IL-6 and IgG in serum;and upregulated splenic levels of CD45+CD3+T lymphocytes and F4/80+CD11b+macrophages,alongside increasing the expression of CD4,CD8 and F4/80 surface markers.CONCLUSION CFM alleviates CTX-induced immunosuppression state in mice by modulating immune cells,restoring immune function and enhancing anti-inflammatory and tissue repair capabilities.
7.Effects of Hedysarum polybotrys polysacchcaide on FXR-FGF19 signal pathway in diabetes rats
Lei ZHANG ; Sheng-fang WAN ; Ya-ling LI ; Qian-kun LIANG ; Yi-hong TIAN ; Xin-xin MA ; Qian GUO
The Chinese Journal of Clinical Pharmacology 2025;41(1):76-80
Objective To study the effects of Hedysarum polysaccharides polysaccharide(HPS)on the farnesoid X receptor(FXR)-fibroblast growth factor-19(FGF19)signaling pathway of diabetes rats.Methods Twelve Wistar male rats were randomly selected as the normal group,and the other rats were fed with a single intraperitoneal injection of streptozotocin(50 mg·kg-1 STZ)and a high sugar and high-fat diet to replicate the diabetes rat model.Model rats were randomly divided into model group,positive control group(given 400 mg·kg-1·d-1 suspension of Bifidobacterium quadruplex live bacterial tablets by gavage),experimental-H,-M,-L groups(given 200,100,and 50 mg·kg-1·d-1 doses of HPS suspension by gavage);normal group,and model group were given equal volume of purified water by gavage once a day for 8 consecutive weeks.Glucose(Glu)was detected by a blood glucose meter;and serum total glyceride(TG)and total cholesterol(TC)were detected by enzyme-linked immunosorbent assay reagent kit;the expressions of FXR、fibroblast growth factor receptors 4(FGFR4)relative mRNA expression level and protein were detected by real-time fluorescence quantitative polymerase chain reaction method and Western blot.Results The Glu concentrations in the normal group,model group,positive control group,and experimental-H groups were(7.66±0.61),(29.25±1.64),(23.31±3.02)and(19.31±5.13)mmol·L-1,respectively;the TG content were(957.00±113.73),(1 345.00±246.44),(958.00±96.53)and(964.00±130.22)μmol·L-1,respectively;the TC content were(161.65±4.53),(302.19±5.35),(236.09±5.14)and(165.58±2.58)μmol·L-1,respectively;the expression of FXR relative mRNA expression level were 1.00±0.06,0.48±0.02,0.67±0.04 and 0.92±0.04,respectively;the expression of FGFR4 relative mRNA expression level were 1.00±0.04,0.17±0.01,0.48±0.04 and 0.41±0.03;respectively.The above indexes of the model group were compared with the control group,and the above indexes of the control group and the experimental-H group were compared with the model group,and the differences were statistically significant(all P<0.01).Conclusion HPS improves blood sugar,lowers blood lipids,and protects liver and intestinal tissues,possibly by regulating the FXR-FGF19 signaling pathway in intestinal tissue,and regulating bile acid synthesis.
8.LncRNA SNHG12 Promotes Breast Cancer Progression via Competing with EPHB3 for Binding to miR-326
Yong LI ; Yi-Ning QUAN ; Kun WANG ; Hong-Li LI ; Chong-Gao YIN
Chinese Journal of Biochemistry and Molecular Biology 2025;41(9):1310-1319
Breast cancer(BRCA)remains one of the leading causes of cancer-related deaths worldwide due to its high rates of metastasis and recurrence,making it crucial to explore its underlying molecular mechanisms.Our previous study demonstrated that miR-326 inhibits BRCA progression by targeting EPH receptor B3(EPHB3).This study further explores the molecular mechanism by which long non-coding RNAs(LncRNAs)regulates BRCA progression via the competing endogenous RNA(ceRNA)mecha-nism,in which it competes with EPHB3 for miR-326 binding.Bioinformatics analysis identified LncRNA Small Nucleolar RNA Host Gene 12(SNHG12)as a potential miR-326-binding molecule.SNHG12 was found to be significantly upregulated in BRCA tissues,exhibiting a negative correlation trend with miR-326 and a positive correlation trend with EPHB3,suggesting its potential involvement in the ceRNA regu-latory network.Nuclear-cytoplasmic fractionation assays revealed cytoplasmic localization of SNHG12,while dual-luciferase reporter assays confirmed its direct binding to miR-326.Functional experiments demonstrated that SNHG12 knockdown significantly suppressed BRCA cell proliferation,invasion,and migration,while miR-326 inhibition reversed these effects.Furthermore,miRNA pulldown assay re-vealed significant enrichment of SNHG12 and EPHB3 in the miR-326 pulldown products,indicating di-rect binding between them.Western blotting and rescue experiments revealed that SNHG12 upregulates EPHB3 expression by sponging miR-326,thereby promoting the malignant behaviors of BRCA cells.Col-lectively,this study revealed that LncRNA SNHG12 promotes BRCA progression by regulating the miR-326/EPHB3 axis through a ceRNA mechanism.The SNHG12/miR-326/EPHB3 pathway may represent a promising target for the molecular diagnosis and targeted therapy of BRCA.
9.Improved YOLOv5 algorithm-based research on CT image recognition and segmentation for cerebral hemorrhage
Cheng-kun HONG ; Tao YANG ; Li-yuan FU
Chinese Medical Equipment Journal 2025;46(5):1-8
Objective To modify the YOLOv5 algorithm with similarity attention mechanism(SimAM)to enhance the recognition and segmentation accuracy of CT images for cerebral hemorrhage.Methods A basic framework was established with a YOLOv5 algorithm consisting of a backbone network(Backbone),a neck module(Neck)and a head module(Head),and then SimAM was introduced at the end of Backbone to form a YOLOv5-Sim-B algorithm and at the end of Neck to construct a YOLOv5-Sim-N algorithm.The YOLOv5-Sim-B and YOLOv5-Sim-N algorithms were trained and validated using the CT image dataset for cerebral hemorrhage publicly available on the Kaggle competition platform,and compared with the traditional YOLOv5 algorithm for recognizing and segmenting cerebral hemorrhagic lesions in CT images.Results In case the value of IoU-T was 0.6,the mean average precision(mAP)was 0.967 for YOLOv5-Sim-B algorithm,0.960 for the YOLOv5-Sim-N algorithm and 0.964 for the traditional YOLOv5 algorithm during the recognition and segmentation of cerebral hemorrhagic lesions in CT images.Conclusion The proposed algorithm gains advantages in detection accuracy and robustness,and can efficiently identify and segment cerebral hemorrhage foci in CT images.[Chinese Medical Equipment Journal,2025,46(5):1-8]
10.Impact of ischemia time and storage periods on RNA quality of fresh-frozen breast cancer and esophageal cancer tissue samples in biobank
Yang-si ZHENG ; Xuan-hao LIN ; Fan LI ; Kun-sheng XIAO ; Xi-feng CHEN ; Chun-peng LIU ; Pei-xiu YAO ; Shao-hong WANG
Fudan University Journal of Medical Sciences 2025;52(3):437-445
Objective To investigate the effects of ischemia time and storage periods on RNA quality in fresh-frozen breast cancer(BC)and esophageal cancer(EC)tissue samples in order to establish evidence-based protocols for biobank sample management.Methods The tumor(T)and paired normal(N)tissue samples from 6 cases of BC and 6 cases of EC were collected and cryopreserved in Biobank,Shantou Central Hospital.Mirror paraffin-embedded tissues were simultaneously prepared into sections for morphological analysis.The samples were divided into two groups of<15 min and 15-30 min according to ischemia time,and RNA quality was analyzed at 4 storage periods of 8-10 months(T1),14-16 months(T2),26-28 months(T3)and 38-40 months(T4).Results In 96 analyzed samples,93.8%(90/96)exhibited high quality(RIN≥6),with 89.6%(43/48)in BC and 97.9%(47/48)in EC.Significant differences in RIN were observed between BC group and EC group(8.050 vs.8.600,P=0.009).In EC group,RIN value was significantly negatively correlated with RNA yield(P<0.001).Moreover,RIN values of tumor-normal pairs exhibited markedly significant differences(7.550 vs.9.000,P<0.001).In contrast,no significant difference was detected in BC group(8.200 vs.7.700,P=0.348).Statistical analysis showed that RIN value was positively correlated with 28S/18S(P<0.001),but had no correlation with tumor content(P=0.676)and necrotic content(P=0.055).Neither ischemia time(<15 min vs.15-30 min:8.200 vs.8.300,P=0.932)nor storage periods(T1-T4:8.400,7.700,8.450,8.600,P=0.163)compromised RNA quality.Conclusion Organ origin and tissue type could influence RNA quality of fresh-frozen tissue samples.However,limited ischemia time(≤30 min)and long-term storage period(38-40 months)do not adversely affect RNA quality in fresh-frozen breast cancer and esophageal cancer tissue samples.

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