1.Diagnostic and prognostic value of differential expression of Cyclin D1 and p53 in eyelid tumors
Minli MA ; Jingjing ZHANG ; Ying LIU ; Hongyan SONG ; Fang LIN ; Qi MA
International Eye Science 2026;26(3):511-517
AIM: To investigate the diagnostic and prognostic value of differential expression of Cyclin D1 and p53 in eyelid tumors.METHODS: This retrospective study enrolled patients who underwent surgical resection for eyelid tumors at our hospital between March 2018 and March 2023. Participants were categorized into benign and malignant groups based on tumor characteristics. Clinical data were collected. Genetic data for eyelid tumors were obtained from the GEO database, and differential gene analysis, including volcano plot visualization and KEGG pathway enrichment analysis, was performed using the Sangerbox 3.0 platform. Immunohistochemistry was used to detect the expression levels of Cyclin D1, p53, and BAX in tissue samples. Correlations with clinical features were analyzed using Spearman analysis, and prognostic factors were identified via Logistic regression analysis.RESULTS: This study included 69 patients with eyelid tumors(78 eyes), categorized into a benign group(37 patients, 41 eyes)and a malignant group(32 patients, 37 eyes)based on tumor characteristics. There were significant differences between the two groups in histological subtype, TNM staging, vascular invasion, differentiation status, and local infiltration(all P<0.05). Among benign tumors: pigmented nevi in 11 eyes(27%), hemangiomas in 9 eyes(22%), squamous cell papillomas in 5 eyes(12%), epidermoid cysts in 5 eyes(12%), seborrheic keratoses in 4 eyes(10%), neurofibromas in 3 eyes(7%), and both calcifying epithelioma and xanthelasma in 2 eyes each(5%); among malignant tumors: basal cell carcinoma in 18 eyes(49%), meibomian gland carcinoma in 8 eyes(22%), squamous cell carcinoma in 5 eyes(14%), sebaceous gland carcinoma in 4 eyes(11%), lymphoma and malignant melanoma each in 1 eye(3%). At the follow-up cutoff date of March 2025, the 2-year survival rate in the benign group(95%)was significantly higher than that in the malignant group(78%; P<0.05). Bioinformatics analysis identified 4 103 differentially expressed genes, including Cyclin D1, p53, and BAX, which were predominantly involved in pathways such as the p53 signaling pathway and calcium-related signaling. Spearman analysis revealed that local invasion(rs=0.71, P<0.05)and TNM stage(rs=0.73, P<0.05)correlated with Cyclin D1 expression; local invasion(rs=0.76, P<0.05)and histological subtype(rs=0.65, P<0.05)correlated with p53 expression. Logistic regression results indicated that Cyclin D1, p53, TNM staging, and local invasion were prognostic risk factors. ROC curve analysis demonstrated that the combined detection of these four indicators had the highest predictive value for prognosis(AUC=0.83).CONCLUSION: High expression of cyclin D1 and p53 serves as molecular markers for distinguishing benign from malignant eyelid tumors and assessing prognosis. Combined detection of these markers with TNM staging and local invasion demonstrates high predictive value for prognosis.
2.Exploring Anti-inflammatory Synergistic Mechanism of Atractylodis Macrocephalae Rhizoma Processed with Aurantii Fructus Immaturus Juice Based on Differential Component Tracking Strategy
Hongda XUAN ; Shengnan SHEN ; Linlin LI ; Jingjing LIAO ; Xianyu XU ; Xiaoxia LIU ; Haining LYU ; Fang WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):228-237
ObjectiveTaking Aurantii Fructus Immaturus juice(AFI)-processed Atractylodis Macrocephalae Rhizoma(AMR) as an example, this study aims to systematically compare the volatile and non-volatile components of AMR and its processed products, investigate the key differential components, evaluate their anti-inflammatory activities, and elucidate the synergistic mechanism of processing. MethodsThe chemical compositions of volatile and non-volatile components in AMR and AFI-processed AMR were systematically characterized using gas chromatography-mass spectrometry(GC-MS) and ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS), with relative mass fractions and response values determined separately. Volatile components were identified through searches in the National Institute of Standards and Technology(NIST)17 database, comparison with retention index(RI) and fragmentation pattern matching. Non-volatile components were identified by searching Waters Traditional Chinese Medicine (TCM) spectral library, in conjunction with PubChem and MassBank, characteristic fragmentation patterns and response values were also used to support identification. Differential components were screened using principal component analysis(PCA), orthogonal partial least squares-discriminant analysis(OPLS-DA), with variable importance in the projection(VIP) value >1. Components with high log2fold change(FC) among major differential groups were selected as those exhibiting significant changes before and after processing. The anti-inflammatory activity of the differential compounds was evaluated by assessing their effects on nitric oxide(NO) production in a lipopolysaccharide(LPS)-induced RAW264.7 macrophage model. Enzyme-linked immunosorbent assay(ELISA) was used to detect the effects of the differential components on tumor necrosis factor(TNF)-α, interleukin(IL)-1β, IL-6, and monocyte chemotactic protein(MCP)-1 levels, and immunofluorescence(IF) was employed to assess their effects on nuclear transcription factor(NF)-κB p65 translocation, thereby elucidating the underlying molecular mechanisms. ResultsA total of 36 compounds were identified in the volatile components of AMR and AFI-processed AMR, among which, sesquiterpenes and monoterpenes were significantly increased after processing. In the non-volatile components, 36 compounds were identified, and the main differential components were flavonoids, sesquiterpenoids, and triterpenoids. Flavonoids were the primary differential components distinguishing AMR from its processed products, representing compounds directly introduced during processing. Five compounds, including atractylenolide Ⅲ, tangeritin, nobiletin, hesperidin and narirutin, were selected as representatives of three classes based on their most prominent differential expression among different compound types for subsequent anti-inflammatory activity studies. The results showed that 100 μmol·L-1 tangerine and narirutin could significantly inhibit LPS-induced NO production(P<0.01) in a concentration-dependent manner. Tangeritin was able to significantly inhibit the levels of TNF-α and MCP-1 secreted by RAW264.7(P<0.05), while narirutin significantly inhibited the levels of TNF-α, IL-1β, MCP-1 and IL-6(P<0.01). IF revealed that both tangeritin and narirutin significantly blocked the translocation of NF-κB p65 from the cytoplasm to the nucleus. ConclusionAFI-processed AMR significantly alters the chemical composition profile of AMR, and the newly introduced flavonoid components during processing may be key to its enhanced anti-inflammatory effects.
3.Current status of brucellosis control programmes and Chinese path based on the One Health concept: a review
Kaihui ZHANG ; Fang LUO ; Lei XIU ; Yongzhi WANG ; Jia WANG ; Jingjing LI ; Jian LI ; Wei HU
Chinese Journal of Schistosomiasis Control 2026;38(4):369-378
Brucellosis is one of the most widespread zoonoses globally, which poses serious threats to both global public health and development of animal husbandry. Conventional brucellosis control measures suffer problems of surveillance coordination, regional joint prevention, and policy implementation, which are insufficient to address the challenges of ecological risks and crosshost transmission. The One Health approach allocates interdisciplinary and intersectoral collaborations to achieve systemic health of humans, animals, and environments, which provides new insights into brucellosis control programmes. This review describes the pathogenic biological characteristics, global epidemiological landscape, and transmission ecology of brucellosis, evaluates the main limitations of current control strategies, summarizes the experiences from successful One Health practices across countries at different stages of development, and proposes the brucellosis control strategy based on the One Health concept with adaptations to the current epidemiology of brucellosis in China, so as to provide the theoretical evidence and practical framework for the national brucellosis control programmes in China, thereby facilitating the effective control and the ultimate elimination of brucellosis.
4.miR-27a-3p promotes the proliferation of human hypertrophic scar fibroblasts by regulating mitogen-activated protein kinase signaling pathway
Jun LI ; Jingjing GONG ; Guobin SUN ; Rui GUO ; Yang DING ; Lijuan QIANG ; Xiaoli ZHANG ; Zhanhai FANG
Chinese Journal of Tissue Engineering Research 2025;29(8):1609-1617
BACKGROUND:Multiple studies have confirmed that mitogen-activated protein kinase(MAPK)signaling pathway is involved in cell proliferation,and microRNA(miR)is involved in the occurrence and development of hypertrophic scars.Therefore,the role of miR-27a-3p and MAPK signaling pathways in pathological scar formation has been further explored. OBJECTIVE:To explore the effect of miR-27a-3p on the proliferation of human hypertrophic scar fibroblasts through the MAPK signaling pathway. METHODS:The primary fibroblasts were isolated and collected from the skin samples.The primary fibroblasts were observed by inverted microscope and verified by immunofluorescence.The relative expression level of miR-27a-3p in tissues was detected by qRT-PCR.The target genes of hsa-miR-27a-3p were predicted using the database,and then the predicted target genes were enriched by gene ontology function analysis and biological pathway enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes.There were seven groups:blank control,negative control,miR-27a-3p mimic,miR-27a-3p inhibitor,miR-27a-3p mimic+p38 MAPK inhibitor,miR-27a-3p mimic+extracellular regulated protein kinase inhibitor,miR-27a-3p mimic+c-Jun N-terminal kinase inhibitor.Western blot was used to detect the levels of extracellular regulated protein kinase,c-Jun N-terminal kinase inhibitor.and p38 kinase and their phosphorylation levels.Cell counting kit-8 and EdU were used to detect cell proliferation. RESULTS AND CONCLUSION:Compared with normal skin fibroblasts,hypertrophic scar fibroblasts had stronger proliferative activity(P<0.05)and faster proliferation level(P<0.001).Compared with normal skin,miR-27a-3p was highly expressed in hypertrophic scars(P<0.001).Compared with the negative control group,overexpression of miR-27a-3p could promote cell proliferation activity(P<0.001)and proliferation levels(P<0.001).Compared with the negative control group,knockdown of miR-27a-3p could inhibit the proliferation activity(P<0.05)and proliferation levels(P<0.001).Compared with the negative control group,overexpression of miR-27a-3p promoted the phosphorylated levels of extracellular regulated protein kinase,c-Jun N-terminal kinase,and p38 mitogen-activated protein kinase(P<0.05).Compared with the negative control group,knockdown of miR-27a-3p inhibited the phosphorylated levels of extracellular regulated protein kinase,c-Jun N-terminal kinase,and p38 MAPK(P<0.05).Compared with the miR-27a-3p mimic group,specific inhibitors of extracellular regulated protein kinase,c-Jun N-terminal kinase,and p38 MAPK reversed the effects of miR-27a-3p on the proliferative activity(P<0.01)and proliferation level(P<0.001)of fibroblasts.To conclude,these results suggest that miR-27a-3p promotes the proliferation of human hypertrophic scar fibroblasts by activating the MAPK signaling pathway.
5.Clinical efficacy and safety of a domestic calcipotriol/betamethasone dipropionate ointment in the treatment of stable plaque psoriasis: a multicenter, randomized, double-blind, controlled study
Lixin XIA ; Guang XIANG ; Qingchun DIAO ; Kun HUANG ; Shoumin ZHANG ; Shanshan LI ; Yumei LI ; Zhiqiang SONG ; Qing SUN ; Xiumin YANG ; Meng PAN ; Yuling SHI ; Shuping GUO ; Huiping WANG ; Tiechi LEI ; Xiaoyong ZHOU ; Songmei GENG ; Suchun HOU ; Juan SU ; Yong CUI ; Rixin CHEN ; Yanyan FENG ; Hongxia FENG ; Rushan XIA ; Zudong MENG ; Fang YIN ; Jingjing WANG ; Xinghua GAO
Chinese Journal of Dermatology 2025;58(11):1020-1026
Objective:To evaluate the clinical equivalence between a domestic calcipotriol/betamethasone dipropionate ointment and the originator product in the treatment of stable plaque psoriasis.Methods:A multicenter, randomized, double-blind, three-arm, parallel-group, active- and placebo-controlled study was conducted, and 449 patients aged 18 - 65 years with stable plaque psoriasis were enrolled from 25 hospitals (such as the First Affiliated Hospital of China Medical University). Eligible patients had a baseline physician's global assessment (PGA) score of ≥ 3 points, baseline body surface area (BSA) involvement of 5% - 30%, and a target lesion psoriasis area and severity index (TL-PASI) for plaque elevation of ≥ 3 points. Participants were randomly assigned in a 2:2:1 ratio to the test group ( n = 179), reference group ( n = 180), and placebo group ( n = 90), and applied the domestic calcipotriol/betamethasone dipropionate ointment, originator product, and ointment base respectively, once daily in the evening for 4 weeks. Efficacy and safety were assessed at weeks 1, 2, and 4. The primary efficacy endpoints were the treatment success rates and clinical success rates in each group at week 4. The per-protocol set (PPS) was used for the primary efficacy analysis, and the intention-to-treat (ITT) set for supplementary efficacy analysis. Equivalence between the test and reference preparations was tested using the Cochran-Mantel-Haenszel method adjusted for randomization strata. Superiority of the test and reference preparations over the placebo was also tested. Measurement data were compared among the 3 groups using analysis of variance or non-parametric tests, while treatment success rates, clinical success rates, and incidence rates of adverse reactions were compared using the chi-square test. Results:The ITT, PPS, and safety sets included 447, 420, and 448 patients, respectively. In the ITT set, patients were aged 43.6 ± 12.8 years, including 320 (71.6%) males and 127 (28.4%) females, and the disease duration was 11.21 ± 9.05 years; 316 (70.7%) had a PGA score of 3 points and 131 (29.3%) had a PGA score of 4 - 5 points. No significant differences in the baseline characteristics (including age, sex, disease duration and disease severity) were observed among the 3 groups (all P > 0.05). Based on the PPS analysis, the treatment success rates were 57.9% (99/171) in the test group, 50.3% (86/171) in the reference group, and 7.7% (6/78) in the placebo group, and the clinical success rates were 57.9% (99/171), 50.3% (86/171), and 10.3% (8/78), respectively; both the test and reference groups were superior to the placebo group in both treatment and clinical success rates (all P < 0.001) ; the rate differences for treatment success (90% confidence interval [ CI]: -1.3% - 16.4%) and clinical success (90% CI: -1.3% - 16.3%) between the test and reference groups were entirely within the pre-defined equivalence margin (-20% - 20%). Subgroup analyses by baseline PGA scores: for patients with a baseline PGA score of 3 points, the treatment success rates in the test, reference, and placebo groups were 60.8% (73/120), 52.1% (62/119), and 11.1% (6/54), respectively, and the corresponding clinical success rates were 61.7% (74/120), 53.8% (64/119), and 13% (7/54), respectively; the test and reference groups did not differ significantly in treatment or clinical success rates (both P > 0.05), but both showed higher success rates than the placebo group (all P < 0.001) ; the results of statistical comparisons among the 3 groups in patients with a baseline PGA score of 4 - 5 points were consistent with those observed in patients with a baseline PGA score of 3 points. The percentage reductions in PGA and TL-PASI scores from baseline to weeks 1, 2, and 4 showed significant differences among the 3 groups, which were significantly higher in the test and reference groups than in the placebo group (all P < 0.001), but did not differ between the test and reference groups (all P > 0.05). The primary adverse reactions were local skin reactions, such as pruritus, pain, and erythema. The incidence rates of adverse reactions were 8.9% (16/179) in the test group, 7.3% (13/179) in the reference group, and 7.8% (7/90) in the placebo group, with no significant difference among the 3 groups ( P > 0.05) . Conclusions:The domestic calcipotriol/betamethasone dipropionate ointment demonstrated clinical equivalence to the originator product in the treatment of stable plaque psoriasis, and the two agents exhibited comparable efficacy for patients with varying degrees of disease severity, and were comparable in the speed and degree of clinical improvement, with similar favorable safety profiles.
6.Influence of different positive end-expiratory pressure on ventilation of laryngeal mask airway in pediatric patients undergoing laparoscopic surgery: an ultrasound assessment
Jingjing WEI ; Yao CHEN ; Fang GUO ; Cunjin WANG ; Ju GAO
Chinese Journal of Anesthesiology 2025;45(2):184-188
Objective:To evaluate the influence of different positive end-expiratory pressure on the ventilation of laryngeal mask airway in pediatric patients undergoing laparoscopic surgery through ultrasound assessment.Methods:In this randomized controlled trial, 90 pediatric patients of both sexes, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, aged 2-10 yr, with a body mass index of 12-22 kg/m 2, scheduled for elective laparoscopic surgery, were divided into 3 groups ( n=30 each) by a random number table method: group P0, group P3, and group P5. Each group adopted the pressure-controlled mode with an inspiration/expiration ratio of 1∶2, a flow rate of 2 L/min, and a respiratory rate of 16-26 breaths/min. The positive end-expiratory pressure was set at 0, 3 and 5 cmH 2O(1 cmH 2O=0.098 kPa) in P0, P3 and P5 groups respectively. The ultrasonic scores and misalignment rate of the laryngeal mask airway were recorded every 5 min following the pneumoperitoneum, and the occurrence of intraoperative hypoxemia (SpO 2 < 92%) and P ETCO 2 ≥ 45 mmHg(1 mmHg=0.133 kPa) was recorded. The development of hypoxemia, blood staining on the laryngeal mask airway and hoarseness after laryngeal mask airway removal were also recorded. Results:Compared with P0 group, the ultrasound scores were significantly increased at 15 min of pneumoperitoneum in P3 group ( P<0.05), and the ultrasound scores and the laryngeal mask airway misplacement rate were significantly increased, and no significant change was found in the other parameters in P5 group ( P<0.05). There were no statistically significant differences in each parameter between P5 group and P3 group ( P>0.05). There was no statistically significant difference in the incidence of postoperative complications among the three groups ( P>0.05). Conclusions:The laryngeal mask airway provides better ventilatory effect in pediatric patients undergoing laparoscopic surgery when the PEEP is set at 3 cmH 2O.
7.Effects of continuous positive airway pressure on maternal and neonatal outcomes in pregnant women with obstructive sleep apnea syndrome
Zelin TU ; Rui BAI ; Linyan ZHANG ; Jingyu WANG ; Shenda HONG ; Jingjing YANG ; Jun WEI ; Yan WANG ; Yanan LIU ; Xiaosong DONG ; Fang HAN ; Guoli LIU
Chinese Journal of Obstetrics and Gynecology 2025;60(3):171-176
Objective:To analyze the effect of continuous positive airway pressure (CPAP) on maternal and neonatal outcomes in pregnant women with obstructive sleep apnea syndrome (OSAS), especially on the incidence of hypertensive disorder in pregnancy (HDP) in women with moderate to severe OSAS.Methods:A total of 180 pregnant women with OSAS who were diagnosed through sleep monitoring during pregnancy due to high-risk factors of OSAS and registered in Peking University People′s Hospital from January 2021 to May 2024 were selected as the study subjects. Clinical data were collected from medical records for retrospective analysis. According to whether they received standardized treatment with CPAP, they were divided into the CPAP treatment group (42 cases) and the control group (138 cases). The CPAP treatment group consisted of 9 pregnant women with moderate to severe OSAS, while the control group consisted of 34 pregnant women with moderate to severe OSAS. The maternal and neonatal outcomes, the incidence of HDP, placental weight after delivery and placental weight/neonatal birth weight ratio were compared between the two groups.Results:(1) The average gestational age of pregnant women in the CPAP treatment group was higher than that in the control group [(38.7±1.0) vs (38.0±1.4) weeks], the proportion of infants small for gestational age (SGA) in the CPAP treatment group was lower [0 (0/42) vs 12.3% (17/138)], and the birth weight of infants in the CPAP treatment group was bigger [(3 396±475) vs (3 082±710) g); the differences between the two groups were statistically significant (all P<0.05). There were no significant differences between the CPAP treatment group and the control group in terms of delivery mode, rates of postpartum hemorrhage and preterm birth, umbilical artery blood gas analysis pH<7.1, lactate≥6.0 mmol/L, base excess<-12.0 mmol/L and the incidence of gestational diabetes mellitus and HDP (all P>0.05). (2) The placental weight of the CPAP treatment group was significantly lower than that of the control group [(554.0±70.6) vs (615.7±119.1) g], the placental weight/newborn birth weight ratio of the CPAP treatment group was significantly lower than that of the control group (median: 0.17 vs 0.19), and the differences were statistically significant (all P<0.05). (3) The incidence of HDP in pregnant women with moderate to severe OSAS in the CPAP treatment group was lower than that in the control group [1/9 vs 61.8% (21/34)], and the difference was statistically significant ( P<0.05). Conclusions:CPAP treatment could prolong the gestational age in pregnant women with OSAS, reduce the incidence of SGA, increase the birth weight of infants, and reduce the incidence of HDP in pregnant women with moderate to severe OSAS, and is worth promoting in clinical practice. The improvement of neonatal outcomes by CPAP treatment is closely related to the placenta, which is worthy of further exploration.
8.Association of anti-rituximab antibodies with relapse after therapy in children with frequently relapsing or steroid-dependent nephrotic syndrome
Jingjing WANG ; Zhengkun XIA ; Chunlin GAO ; Pei ZHANG ; Tao SUN ; Xiang FANG ; Zhuo SHI ; Ren WANG
Chinese Journal of Pediatrics 2025;63(9):980-984
Objective:To investigate the association between anti-rituximab antibodies (ARA) and relapse after rituximab (RTX) therapy in children with frequently relapsing or steroid-dependent nephrotic syndrome (FRNS or SDNS).Methods:A retrospective cohort study was conducted. Clinical and laboratory data were collected from 48 FRNS or SDNS children treated with RTX in the Department of Pediatrics, General Hospital of Eastern Theater Command, between April 2024 and October 2024. Data included RTX dosing frequency, relapse events, peripheral CD20? B-cell counts, and ARA levels. With a 6-month observation period after the last RTX therapy, the children were divided into an ARA-positive group and an ARA-negative group based on ARA test results. Chi-square test, independent sample t-test, or Mann-Whitney U test were used to compare relapse rates and laboratory indicators between the two groups. The predictive value of ARA levels for relapse was evaluated using univariate receiver operating characteristic (ROC) curve analysis. Results:Among the 48 children (36 males, 12 females), the age of disease onset was 3.5 (2.0, 6.0) years, the ages at the first and last RTX treatments were 7.0 (5.0, 12.0) years and 9.5 (7.0, 13.0) years, respectively. The overall ARA positive rate was 29% (14/48). The relapse rate in the ARA-positive group was significantly higher than that in the negative group ( P<0.05). The ARA level was 0.01 (0.01, 5.88) μg/L, and all 12 children with ARA levels >5.88 μg/L relapsed. ROC curve analysis showed that ARA levels predicted relapse after RTX treatment in FRNS or SDNS children with an area under the curve (AUC) of 0.73, sensitivity of 0.50, specificity of 1.00, and an optimal cut-off value of 5.02 μg/L. All children received single-dose RTX therapy, with no significant difference in treatment frequency between the two groups ( P>0.05). At 3 months after the last rituximab therapy, CD20? B cell counts were significantly higher in the ARA-positive group ( P<0.05). During follow-up, 15% (7/48) of the children experienced infusion-related adverse reactions, with no significant difference in incidence between the two groups ( P>0.05). Conclusion:ARA is significantly associated with relapse in FRNS or SDNS children after RTX therapy.
9.Construction and practice of smart health and elderly care standard system in Shanghai
Jian WANG ; Mianzhi CHENG ; Xiaohua YE ; Weihua GU ; Chun FAN ; Yuyao JIANG ; Min XU ; Yihan XU ; Yang WANG ; Xiaoyan GU ; Yihua JIANG ; Liying YAO ; Shusheng OUYANG ; Xin LIU ; Xijie YUAN ; Jian CHEN ; Ni YANG ; Qi CHEN ; Jingjing FANG
Journal of Navy Medicine 2025;46(1):83-90
With the rapid development of population aging in various countries around the world,the health and elderly care industry has been paid high attention.The standardization of smart health and elderly care technology and services is particularly important.This paper firstly reviewed the policies related to healthy elderly care in China.By analyzing the industrial standards and provincial standards issued,this paper focused on the policies proposed by the Shanghai Municipal Government for the standardization of smart health and elderly care,as well as the researches on the standard system and the construction of standard families.Shanghai group standards in the field of smart health and elderly care were summarized,including the guidelines for the construction of standard systems,elderly care service platforms,community elderly cafeterias,portable health monitoring terminals,indoor sports services,and home-based elderly care safety monitoring.A series of case analyses of the standardized implementation of the above aspects were also provided.Through standardization research and practice in recent years,it has been fully demonstrated that the standard research plays an important leading role in the field of smart health and elderly care.
10.Study on mechanism of immunogenic cell death induced by ginsenoside octanoate through induction of autophagy in hepatocellular carcinoma cells
Fuxiang SONG ; Zhenzhen DAI ; Jingjing SHENG ; Jiali CHEN ; Hui ZHANG ; Hua FENG ; Yao PAN ; Zeyuan DENG ; Fang CHEN
Chinese Journal of Immunology 2025;41(6):1427-1432
Objective:To investigate the effect of ginsenoside octanoate(Rh2-O)on inducing immunogenic cell death in hepa-tocellular carcinoma cells and its molecular mechanism.Methods:Effects of ginsenoside caprylate(Rh2-O)and autophagy inhibitor 3-MA on the activity of hepatocellular carcinoma cells were detected by CCK-8 assay.The effect of Rh2-O on CRT membrane eversion in Hepa1-6 cells were detected by immunofluorescence assay.Rh2-O treated mouse hepatocellular carcinoma cells were used to pre-pare a tumor vaccine for in vivo vaccination experiments in mice.Extracellular ATP levels were detected in real-time.The expression of autophagy-related genes and proteins were measured by real-time fluorescence PCR and Western blot,and the mitochondrial morphol-ogy and co-localization with autophagy proteins were observed by laser confocal microscopy.Results:Rh2-O showed strong cytotoxicity to Hepa1-6 cells[cell viability:(58.54±3.56)%]at a concentration of 150 μmol/L,and a large amount of CRT was observed on the surface of the cell membrane.The tumor emergence rate was 36.36%in the vaccinated group and 100%in the control group.The tumor vaccine prepared by Rh2-O effectively protected mice from the same type of tumor attack;Rh2-O induced an increase in the level of cellular secreted ATP(P<0.05),the mRNA of autophagy-related genes ATG3,p62,LC3 expression levels and autophagy-associated proteins LC3A and LC3B expression levels were increased(P<0.05),and co-localization of mitochondria with autophagy proteins was significantly increased(P<0.05).In addition,Rh2-O action on 3-MA pretreated hepatocellular carcinoma cells resulted in a signifi-cant decrease in extracellular ATP levels(P<0.001).Conclusion:Rh2-O may induce immunogenic cell death by inducing autophagy in hepatocellular carcinoma cells.

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