1.Correlation Analysis of Huanglian Jiedu Wan on Syndrome Improvement and Clinical Biomarkers of "Excess Heat-Toxicity" Based on Machine Learning Model
Qi LI ; Keke LUO ; Baolin BIAN ; Hongyu YU ; Mengxiao WANG ; Mengyao TIAN ; Wen XIA ; Yuan MA ; Xinfang ZHANG ; Pengyue LI ; Nan SI ; Hongjie WANG ; Yanyan ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):162-173
ObjectiveThis paper aims to find the identified and validated clinical biomarker data building upon a clinical study of early-phase phase Ⅱ and investigate the correlation analysis of Huanglian Jiedu Wan on syndrome improvement and clinical biomarkers in the treatment of "excess heat-toxicity" based on a machine learning model. Additionally, the effective prediction of clinical biomarker values for the main symptoms of the "excess heat-toxicity" syndrome was assessed. MethodsA total of 229 patients meeting the inclusion criteria for "excess heat-toxicity" syndrome were randomly divided into the Huanglian Jiedu Wan group and the placebo group. Syndrome score transition matrices were constructed for the Huanglian Jiedu Wan group and the placebo group based on three main symptoms of "excess heat-toxicity" syndrome, such as oral ulcers, sore throat, and gum swelling and pain. Data from the patients with these three syndromes were also integrated for an overall analysis. The corresponding syndrome score transition matrices were further constructed to visualize symptom change trends of the patients in the two groups via heatmaps. Based on the identified and validated clinical biomarkers related to inflammation, oxidative stress, and energy metabolism in the early phase, Spearman correlation analysis was employed to analyze and evaluate the associations between clinical biomarkers and syndrome improvement. Key clinical biomarkers reflecting the effect of Huanglian Jiedu Wan were screened through the comparison of differences between groups. An extreme gradient boosting (XGBoost) algorithm was used to develop a prediction model for main symptom classification, with classification performance evaluated through 10-fold cross-validation. Feature importance analysis was applied to identify variables with the greatest contribution to the prediction result. ResultsThe syndrome transition matrix results indicated that the Huanglian Jiedu Wan group showed a superior effect to the placebo group in improving oral ulcers, sore throat, and overall symptoms, with significant effects observed especially in sore throat and overall symptom analyses (P<0.01). Spearman correlation analysis revealed that several clinical biomarkers positively correlated with "excess heat-toxicity" syndrome and its main symptom improvement, were also called "heat-related biomarkers", including succinic acid, α-ketoglutaric acid, glycine, lactic acid, adenosine monophosphate (AMP), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-1β (IL-1β), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), and so on. Conversely, clinical biomarkers negatively correlated with symptom severity, were also called "heat-clearing related biomarkers" after administration of Huanglian Jiedu Wan, including malic acid, fumaric acid, cis-aconitic acid, adrenocorticotropic hormone (ACTH), IL-1β, IL-4, IL-8, succinic acid, and citric acid. The XGBoost classification model using all 52 biomarkers as variables achieved an average test accuracy of 0.754 and an average F1 score of 0.777. Feature importance analysis identified the scores of glutamic acid in saliva and IL-6 were the highest in all the variables, with importance scores of 0.081 and 0.080, respectively. After screening out 14 key variables and optimizing the parameters, model performance improved to an average accuracy of 0.758 and an F1 score of 0.798. Feature importance analysis further determined that the glutamic acid in saliva and IL-6 showed obvious changes after screening the variables, confirming the good syndrome prediction ability of the model constructed by these key clinical biomarkers. ConclusionThis study systematically elucidates the correlation between syndrome improvement and clinical biomarkers of Huanglian Jiedu Wan in the treatment of "excess heat-toxicity" syndrome. An XGBoost classification model based on key clinical biomarkers is successfully established, achieving effective prediction of the symptoms related to the "excess heat-toxicity" syndrome such as oral ulcers and sore throat and providing a new insight for objective identification of traditional Chinese medicine syndromes.
2.Correlation Analysis of Huanglian Jiedu Wan on Syndrome Improvement and Clinical Biomarkers of "Excess Heat-Toxicity" Based on Machine Learning Model
Qi LI ; Keke LUO ; Baolin BIAN ; Hongyu YU ; Mengxiao WANG ; Mengyao TIAN ; Wen XIA ; Yuan MA ; Xinfang ZHANG ; Pengyue LI ; Nan SI ; Hongjie WANG ; Yanyan ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):162-173
ObjectiveThis paper aims to find the identified and validated clinical biomarker data building upon a clinical study of early-phase phase Ⅱ and investigate the correlation analysis of Huanglian Jiedu Wan on syndrome improvement and clinical biomarkers in the treatment of "excess heat-toxicity" based on a machine learning model. Additionally, the effective prediction of clinical biomarker values for the main symptoms of the "excess heat-toxicity" syndrome was assessed. MethodsA total of 229 patients meeting the inclusion criteria for "excess heat-toxicity" syndrome were randomly divided into the Huanglian Jiedu Wan group and the placebo group. Syndrome score transition matrices were constructed for the Huanglian Jiedu Wan group and the placebo group based on three main symptoms of "excess heat-toxicity" syndrome, such as oral ulcers, sore throat, and gum swelling and pain. Data from the patients with these three syndromes were also integrated for an overall analysis. The corresponding syndrome score transition matrices were further constructed to visualize symptom change trends of the patients in the two groups via heatmaps. Based on the identified and validated clinical biomarkers related to inflammation, oxidative stress, and energy metabolism in the early phase, Spearman correlation analysis was employed to analyze and evaluate the associations between clinical biomarkers and syndrome improvement. Key clinical biomarkers reflecting the effect of Huanglian Jiedu Wan were screened through the comparison of differences between groups. An extreme gradient boosting (XGBoost) algorithm was used to develop a prediction model for main symptom classification, with classification performance evaluated through 10-fold cross-validation. Feature importance analysis was applied to identify variables with the greatest contribution to the prediction result. ResultsThe syndrome transition matrix results indicated that the Huanglian Jiedu Wan group showed a superior effect to the placebo group in improving oral ulcers, sore throat, and overall symptoms, with significant effects observed especially in sore throat and overall symptom analyses (P<0.01). Spearman correlation analysis revealed that several clinical biomarkers positively correlated with "excess heat-toxicity" syndrome and its main symptom improvement, were also called "heat-related biomarkers", including succinic acid, α-ketoglutaric acid, glycine, lactic acid, adenosine monophosphate (AMP), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-1β (IL-1β), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), and so on. Conversely, clinical biomarkers negatively correlated with symptom severity, were also called "heat-clearing related biomarkers" after administration of Huanglian Jiedu Wan, including malic acid, fumaric acid, cis-aconitic acid, adrenocorticotropic hormone (ACTH), IL-1β, IL-4, IL-8, succinic acid, and citric acid. The XGBoost classification model using all 52 biomarkers as variables achieved an average test accuracy of 0.754 and an average F1 score of 0.777. Feature importance analysis identified the scores of glutamic acid in saliva and IL-6 were the highest in all the variables, with importance scores of 0.081 and 0.080, respectively. After screening out 14 key variables and optimizing the parameters, model performance improved to an average accuracy of 0.758 and an F1 score of 0.798. Feature importance analysis further determined that the glutamic acid in saliva and IL-6 showed obvious changes after screening the variables, confirming the good syndrome prediction ability of the model constructed by these key clinical biomarkers. ConclusionThis study systematically elucidates the correlation between syndrome improvement and clinical biomarkers of Huanglian Jiedu Wan in the treatment of "excess heat-toxicity" syndrome. An XGBoost classification model based on key clinical biomarkers is successfully established, achieving effective prediction of the symptoms related to the "excess heat-toxicity" syndrome such as oral ulcers and sore throat and providing a new insight for objective identification of traditional Chinese medicine syndromes.
3.Statistical analysis of vector monitoring in port areas of Shandong Province from 2017 to 2023
Huan-mei HAN ; Tao ZHANG ; Zhi-ping SU ; Rong-jun YAN ; Wei HUANG ; Wen-wen ZHANG
Acta Parasitologica et Medica Entomologica Sinica 2026;33(2):121-127
Objective To provide a scientific basis for vector prevention and control in port areas, vector monitoring has been conducted continuously to characterize the composition, density, and seasonal fluctuations of vectors across 21 port-areas of Shandong province from 2017 to 2023. Methods In accordance with the Regional Vector Monitoring Plan for Frontier Ports, vector surveillance of mosquitoes, flies, cockroaches, rodents, and other disease vectors was performed. Data were analyzed to describe vector community composition and identify seasonal fluctuations. Results A total of 601 rodents belonging to 1 family, 4 genera, and 4 species were captured. Three dominant rodent species inhabit port areas. In addition, 88 104 mosquitoes were captured and classified into 3 subfamilies, 4 genera, and 12 species; 38 959 flies were trapped and classified into 8 families,31 genera, and 51 species, including 9 dominant fly species in port-areas; and 54 672 captured cockroaches were classified into 2 families,4 genera, and 4 species, with Blattella germanica as the dominant species. Conclusions Significant vector community changes(P<0.005)have been identified after the epidemic. To improve the pertinence and efficacy of port health quarantine work, continuous and strong attention should be paid to vector compositions and population densities at Shandong ports.
4.Programmed Cell Death in Endometriosis and Traditional Chinese Medicine Intervention: A Review
Zuoliang ZHANG ; Wanrun WANG ; Wen LI ; Xue HAN ; Xiaohong CHEN ; Nan SU ; Huiling LIU ; Quansheng WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):48-57
Endometriosis (EMT) is a common disease with frequent occurrence and difficult to be cured in modern clinical practice of obstetrics and gynaecology. It is characterized by progressively worsening dysmenorrhoea, pelvic mass, and infertility. The incidence of EMT is growing and increasingly younger patients are diagnosed with this disease, which poses a serious threat to the reproductive and psychological health of women of childbearing age and adolescent females. However, the pathogenesis of EMT is still not completely clear, and the disease has a long course. Therefore, developing new therapies is an urgent clinical problem to be solved. Great progress has been achieved in the treatment of EMT with traditional Chinese medicine (TCM), while the underlying mechanism remains in exploration. Programmed cell death (PCD) is a cell death mode mediated by a variety of bio-molecules with specific signaling cascades. The known PCD processes include apoptosis, pyroptosis, autophagy, ferroptosis, and cuproptosis, which all play a pivotal role in the development of EMT. Researchers have made achievements in the treatment of EMT with TCM, which regulates PCD via multiple pathways, routes, targets, and mechanisms. However, the progress in the regulation of PCD in the treatment of EMT with TCM remains to be reviewed. This paper reviews the research progress in the treatment of EMT with TCM from five PCD processes (apoptosis, pyroptosis, autophagy, ferroptosis, and cuproptosis), with the aim of providing a theoretical basis for the clinical prevention and treatment of EMT.
5.Cross-sectional survey of healthcare-associated infection in 5 736 medical institutions across China in 2024
Cui ZENG ; Wuqiang GAO ; Fu QIAO ; Hui ZHAO ; Xu FANG ; Linping LI ; Xiuwen CHEN ; Jiansen CHEN ; Dan LI ; Yuan ZHOU ; Lingli YU ; Qinglan MENG ; Xia MOU ; Lijuan XIONG ; Weiguang LI ; Ding LIU ; Jiaqing XIAO ; Limei OU ; Baozhen LI ; Jun YIN ; Haojun ZHANG ; Qiang FU ; Qun LU ; Biao WU ; Ya-wei XING ; Shumei SUN ; Shuncai WANG ; Longmin DU ; Jingping ZHANG ; Wen-ying HE ; Gui CHENG ; Nan REN ; Xun HUANG ; Anhua WU
Chinese Journal of Infection Control 2025;24(11):1572-1583
Objective To understand the current situation of healthcare-associated infection(HAI)in China,pro-vide data support and decision-making basis for formulating scientific and effective strategies for HAI prevention and control.Methods A nationwide cross-sectional survey on HAI was conducted among various types and levels of medical institutions in China according to a unified protocol of bedside surveys and case investigations.Results In 2024,a total of 5 736 medical institutions and 2 751 765 patients were surveyed.Among them,34 889 HAI cases were identified,with a prevalence rate of 1.27%.The number of HAI episodes was 38 032,and case prevalence rate was 1.38%.The prevalence rate of HAI in medical institutions in different regions of China ranged from 0.66%to 2.35%.Among medical institutions of different scales,those with a bed capacity of ≥900 had the high-est incidence of HAI,reaching 1.65%.The most common infection site was the lower respiratory tract(44.66%),followed by the urinary tract(12.94%),surgical site(9.32%),upper respiratory tract(7.02%),and bloodstream infection(5.78%).The top 3 departments with the highest HAI rates were the general intensive care unit(10.02%),department of neurosurgery(5.51%),and department(group)of hematology(5.34%).A total of 23 238 strains of HAI pathogens were detected,with 10 714 strains(46.10%)from lower respiratory tract speci-mens.The top 5 detected strains were Klebsiella pneumoniae(14.76%),Pseudomonas aeruginosa(13.33%),Escherichia coli(12.79%),Acinetobacter baumannii(9.23%),and Staphylococcus aureus(7.88%).231 944 pa-tients underwent class Ⅰ incision surgery were monitored,with 1 647 cases experienced surgical site infection,and the prevalence rate of surgical site infection was 0.71%.The number of patients who should undergo pathogen de-tection(patients receiving therapeutic and therapeutic combined prophylactic antimicrobial agents)was 715 179,while the actual number was 480 492,with a pathogen detection rate of 67.18%.425 225 patients received patho-genic detection before treatment,with a detection rate of 59.46%.Conclusion The overall HAI prevalence in Chi-na is lower,showing disparities among medical institutions of different regions and scales.Therefore,precise imple-mentation of measures is necessary for HAI prevention and control,with a focus on high-risk institutions and high-risk departments,key areas,and critical procedures.All levels of medical institutions should continuously reduce the incidence of HAI by strengthening monitoring,standardizing the use of antimicrobial agents,and reinforcing basic HAI prevention and control measures.
6.Clinical characteristics of Mycoplasma pneumonia infection in children of different ages
Nan WU ; Siyang REN ; Wen ZHANG ; Weichao CHEN
Chinese Journal of Primary Medicine and Pharmacy 2025;32(7):975-980
Objective:To investigate the clinical characteristics of Mycoplasma pneumonia infection in children of different ages, and to provide a basis for precise treatment. Methods:A total of 216 children with Mycoplasma pneumonia infection who were admitted to the Xi 'an Children's Hospital from January 2019 to December 2021 were included in this study. These children consisted of 117 males and 99 females. These children were divided into school-age group (> 6 years old, n = 75), preschool group (3-6 years old, n = 72) and infant and toddler group (< 3 years old, n = 69) according to age. Data on general demographics, clinical manifestations, complications, laboratory indicators, imaging findings, and bronchoscopic results, as well as treatment efficacy and outcomes, were collected and compared among the three groups. Results:There were statistically significant differences among the school-age, preschool, and infant and toddler groups in terms of dry cough [45 (60.00%), 34 (47.22%), 18 (26.08%)], wheezing [13 (17.33%), 19 (26.39%), 41 (59.42%)], concurrent pleural effusion [26 (34.67%), 20 (27.78%), 11 (15.94%)], pulmonary necrosis [12 (16.00%), 3 (4.17%), 0 (0)], pulmonary imaging findings (patchy shadows [12 (16.00%), 22 (30.56%), 46 (66.67%)], ground-glass opacities [11 (14.67%), 18 (25.00%), 40 (57.97%)], consolidation shadows [58 (77.33%), 42 (58.33%), 8 (11.59%)]), fever duration [(9.58 ± 4.85) days, (9.48 ± 4.89) days, (6.58 ± 3.64) days], and cough relief time [9 (8,12) days, 9 (8,11) days, 8 (7,11) days] ( χ2 = 16.94, 31.10, 6.59, 15.53, 41.51, 33.40, 65.12, F = 11.97, H = 6.05, all P < 0.05). However, there were no statistically significant differences in fever, dyspnea, concurrent atelectasis, or prognosis ( χ2 = 0.21, 0.27, 0.61, 1.74, all P > 0.05). The percentage of neutrophils [(63.91 ± 10.96)%, (58.26 ± 13.79)%, (50.98 ± 13.79)%], platelet count [(305.01 ± 96.13) × 10 9/L, (324.91 ± 108.05) × 10 9/L, (342.41 ± 120.50) × 10 9/L], erythrocyte sedimentation rate [(47.07 ± 26.46) mm/h, (48.29 ± 26.33) mm/h, (38.16 ± 18.23) mm/h], creatinine [(39.10 ± 7.02) μmol/L, (31.50 ± 5.43) μmol/L, (25.85 ± 4.57) μmol/L], alanine aminotransferase [14 (11, 21) U/L, 12 (9, 20) U/L, 15 (11, 19) U/L], creatine kinase isoenzyme [17 (14, 21) U/L, 20 (16, 24) U/L, 23 (19, 27) U/L], and lactate dehydrogenase [260.0 (224.5, 343.5) U/L, 294.5 (252.0, 379.3) U/L, 317.0 (266.5, 384.5) U/L] levels in the peripheral blood differed significantly among the school-age, preschool, and infant and toddler groups ( F = 37.07, 4.91, 3.55, 167.22, H = 7.54, 57.34, 33.58, all P < 0.05). However, there were no statistically significant differences in peripheral blood white blood cell count, C-reactive protein, or procalcitonin levels ( H = 1.09, 2.49, 2.21, all P > 0.05). The bronchoscopic examinations revealed mucosal congestion and edema in all three groups ( χ2 = 0.51, P > 0.05). However, the detection of lymphoid follicles [43 (57.33%), 28 (38.89%), 18 (26.09%)], longitudinal folds [58 (77.33%), 34 (47.22%), 10 (14.49%)], mucus plugs [46 (61.33%), 32 (44.44%), 6 (8.70%)], and airway shaping [16 (21.33%), 5 (6.94%), 0 (0.00%)] increased with age, showing statistically significant differences among the three groups ( χ2 = 14.72, 56.94, 43.30, 19.58, all P < 0.05). Conclusions:The clinical characteristics of children with Mycoplasma pneumonia infection vary among children of different ages. In infants and young children, wheezing symptoms are more common, and they are prone to multiple organ dysfunction. Lung imaging primarily shows scattered patchy shadows and ground-glass opacities. In contrast, older children mainly present with dry cough, and lung imaging typically reveals large areas of consolidation. Bronchoscopic examinations reveal characteristic findings such as lymphoid follicles, longitudinal folds, mucus plugs, and airway shaping, with longer durations of fever and cough relief.
7.Design and performance verification of high altitude adaptive oxygen generator
Bo WANG ; Xiao-feng LIU ; Wen-jia LIU ; Yi LI ; Ya-nan WU ; Shu-jie CUI ; Wei ZHANG
Chinese Medical Equipment Journal 2025;46(4):29-34
Objective To design a high altitude adaptive oxygen generator for the crews to alleviate their high altitude reaction in high altitude environment and meet their requirements for oxygen supply.Methods A high altitude adaptive oxygen generator based on the mature pressure swing adsorption oxygen production method was designed with the key technologies of discharge capacity compensation of air compression pump and airway fusion of molecular sieve tower,which had the components of molecular sieve tower,air compression pump,controller,cooling fan,cooler,solenoid valve,regulator,flow meter and etc.Trials were carried out at the simulated altitude and field plateau environment so as to verify the high altitude adaptive performance of the oxygen generator developed.Results The trial results showed the oxygen generator met the desired objectives and the requirements for oxygen volume fraction in GJB 2799-1996 General specification for medical oxygen generator using molecular sieve method.Conclusion The oxygen generartor provides oxygen supply effectively for vehicle operators in plateau environments or the ones rushing into the plateau.[Chinese Medical Equipment Journal,2025,46(4):29-34]
8.The protective effect of Gualou Guizhi granules on neuronal injury induced by LPS-activated microglia based on Notch signaling pathway
Xue-zhen LI ; Xiao-xue ZOU ; Wen-ting CHEN ; Yi FENG ; Ya-nan LI ; Yu-qin ZHANG ; Li-hong NAN
Chinese Pharmacological Bulletin 2025;41(4):781-786
Aim To investigate the protective effect of Gualou Guizhi granules(GLGZG)on neuronal injury induced by LPS-activated microglia based on Notch signaling pathway.Methods LPS-activated microglia were co-cultured with neurons to construct neuron inju-ry models,and the cells were divided into the control group,model group,Notch inhibitor(DAPT)group,GLGZG(50,100,200 mg·L-1)group,DAPT+100 mg·L-1GLGZG group.After intervention,the activity of HT22 cells was detected by CCK-8 method,and rel-ative mRNA expression was detected by real-time PCR.The relative protein expression was detected by Western blot.Results Compared with the model group,after GLGZG intervention,the cell activity was significantly improved,GLGZG decreased IL-6,IL-12,Bax,Notch 1,caspase-3,Delta-1,NICD,RBPSUH,HES1 expression,and increased Bcl-2 expression(P<0.05).Compared with the model group,the NICD,RBPSUH and HES1 mRNA and protein expressions significantly decreased after DAPT treatment(P<0.05),and there was no superposition effect with GLG-ZG.Conclusion GLGZG may play a neuroprotective role by inhibiting inflammatory factors and apoptosis,and inhibiting Notch signaling pathway.
9.Monitoring and analysis of the presence of Dabie Bandavirus in rodents in Xilingol League, Inner Mongolia Autonomous Region, China
Xiaowei NAN ; Desu WEN ; Shanshan WANG ; Mengmeng ZHANG ; Xiaoyan SI ; Zhen YAN ; Man LUO ; Haiyang ZHANG ; Zhongbing ZHANG
Chinese Journal of Experimental and Clinical Virology 2025;39(2):149-153
Objective:To analyze the prevalence of the Dabie bandavirus among rats in Taipusi Banner, Xilingol League, Inner Mongolia Autonomous Region.Methods:The cytochrome b gene for the identification of rat species was amplified. Real-Time RT-PCR and RT-PCR were used to amplify and sequence DBV positive rat tissues and detected by XilinGol League Center for Disease Control and Prevention, and the gene evolution was analyzed.Results:The rat was identified as Spermophilus dauricus. Through molecular detection on 20 samples, 15 samples showed positive result in Real Time RT-PCR nucleic acid test, and 7 samples showed positive result in RT-PCR nucleic acid test. Seven base sequences were obtained through sequencing, and they were compared with those in the National Center for Biotechnology Information (NCBI) of the United States by BLAST. It was determined that they were the S gene sequences of DBV. It has been uploaded to GenBank with serial numbers PV231886, PV231887 and PV231888.Conclusions:DBV was detected for the first time in the mice of Inner Mongolia Autonomous Region. In future work, monitoring of DBV carried by rodents and ectoparasites such as ticks and fleas in this area should be strengthened. Publicity and education on severe fever with thrombocytopenia syndrome should be carried out in this area to improve the local medical system′s ability to identify such cases, ensuring early detection, early diagnosis, and early treatment, and reducing the occurrence of human cases and clustered outbreaks.
10.The role of endoplasmic reticulum IP 3R calcium channel in vitamin E succinate induced autophagy of human gastric cancer cell
Xinyi WEN ; Miaomiao CAO ; Zongyue ZHANG ; Nan XIE ; Zhiyang WEI ; Liying HOU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2025;43(3):180-188
Objective:To investigate the role of vitamin E succinate (VES) in inducing autophagy of human gastric cancer cells by activating calcium redistribution through inositol 1, 4, 5-trisphosphate receptors (IP 3R) pathway. Methods:Human gastric cancer lines MKN28 (moderately differentiated) and MKN45 (poorly differentiated) cells were cultured in vitro in March 2022. Gastric cancer cells were treated with VES at different doses for 24 h, and cell viability was measured by CCK-8 method to determine VES dose for subsequent study. The experiment was set up with solvent control group (0.1% ethanol), VES dose groups, 100 nmol/L rapamycin (RAPA) as autophagy positive control group (RAPA group), 15 μg/ml tunicamycin (TM) was used as the endoplasmic reticulum stress (ERS) positive control group (TM group), 10 μmol/ml 2-aminoethyl diphenylborinate (2-APB group) was used to inhibit IP 3R (2-APB group) and VES+2-APB group. The occurrence of autophagosomes in gastric cancer cells was observed by transmission electron microscopy, and microtubule associated protein 1 light chain 3 (LC3), Beclin1, IP 3R, glucose-regulated protein 75 (Grp75), voltage-dependent anion channel 1 (VDAC1) protein expression was detected by western blotting. Fluo-4 AM was used to label intracellular calcium ions, Rhod-2 AM was used to label mitochondrial calcium ions, and the fluorescence intensity of calcium ions was observed by fluorescence microscope. One-way analysis of variance was used to compare the means among multiple groups, and LSD- t method was used for pairwise comparison. Results:CCK-8 results showed that compared with solvent control group, the proliferation rates of MKN28 cells in 10-100 μg/ml VES group and MKN45 cells in 20-100 μg/ml VES group were significantly decreased ( P<0.05). Subsequent VES dosages were determined according to the growth curve, MKN28 was 5, 10, 20, 40 μg/ml, and MKN45 was 10, 20, 40, 80 μg/ml. The results of transmission electron microscopy and fluorescence showed that autophagosomes were formed in MKN28 cells in 5 and 20 μg/ml VES groups and MKN45 cells in 10 and 40 μg/ml VES groups, and the fluorescence intensity of calcium ions in cytoplasm and mitochondria was significantly higher than that in solvent control group ( P<0.05). Compared with solvent control group, LC3, Beclin1, IP 3R, Grp75 and VDAC1 protein expressions of MKN28 cells in 20 and 40 μg/ml VES groups and MKN45 cells in 40 and 80 μg/ml VES groups were significantly increased ( P<0.05). After inhibiting IP 3R with 2-APB, the expression levels of IP 3R, Grp75 and VDAC1 in two kinds of gastric cancer cells in VES+2-APB group were significantly decreased compared with VES group ( P<0.05). The fluorescence results showed that the fluorescence intensity of cytoplasmic and mitochondrial calcium ions in VES+2-APB groups was significantly lower than that in VES group ( P<0.05). Compared with VES group, LC3 and Beclin1 protein expressions in two kinds of gastric cancer cells in VES+2-APB groups were significantly decreased ( P<0.05) . Conclusion:VES may activate intracellular calcium redistribution through IP 3R-Grp75-VDAC1 calcium channel and induce autophagy in gastric cancer cells.


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