1.Application of Ferroptosis Regulation in Chronic Atrophic Gastritis Based on Spleen Deficiency and Turbid Toxin
Yuxi GUO ; Xuemei JIA ; Jie WANG ; Yanru CAI ; Pengli DU ; Yao DU ; Diangui LI ; Qian YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):279-285
Chronic atrophic gastritis (CAG), a common digestive system disease, has an unclear pathogenesis. Currently, it is mostly believed to be related to Helicobacter pylori (Hp) infection, immune factors, dietary factors, bile reflux, long-term use of antibiotics and anti-inflammatory drugs, and other factors. Ferroptosis is a regulated cell death mechanism that is iron-dependent and characterized by disruption of iron metabolism and accumulation of lipid peroxides. More and more studies have found that ferroptosis is closely related to the onset of CAG. Professor LI Diangui, a master of traditional Chinese medicine, first proposed the turbid toxin theory, which holds that spleen deficiency and turbid toxin is the main pathogenic mechanism of CAG. Abnormal iron metabolism regulation is a prerequisite for the accumulation of turbid toxin in CAG, and ferroptosis is in accordance with the pathogenic mechanism (spleen deficiency and turbid toxin) of CAG. This article explores the pathological mechanism of spleen deficiency and turbid toxin in CAG from the perspectives of iron metabolism, oxidative stress, and lipid peroxidation, providing theoretical support of traditional Chinese medicine for the modern research on CAG. It enriches the modern scientific connotation of the turbid toxicity theory and provides new ideas and breakthrough points for the clinical treatment of CAG.
2.Application of Ferroptosis Regulation in Chronic Atrophic Gastritis Based on Spleen Deficiency and Turbid Toxin
Yuxi GUO ; Xuemei JIA ; Jie WANG ; Yanru CAI ; Pengli DU ; Yao DU ; Diangui LI ; Qian YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):279-285
Chronic atrophic gastritis (CAG), a common digestive system disease, has an unclear pathogenesis. Currently, it is mostly believed to be related to Helicobacter pylori (Hp) infection, immune factors, dietary factors, bile reflux, long-term use of antibiotics and anti-inflammatory drugs, and other factors. Ferroptosis is a regulated cell death mechanism that is iron-dependent and characterized by disruption of iron metabolism and accumulation of lipid peroxides. More and more studies have found that ferroptosis is closely related to the onset of CAG. Professor LI Diangui, a master of traditional Chinese medicine, first proposed the turbid toxin theory, which holds that spleen deficiency and turbid toxin is the main pathogenic mechanism of CAG. Abnormal iron metabolism regulation is a prerequisite for the accumulation of turbid toxin in CAG, and ferroptosis is in accordance with the pathogenic mechanism (spleen deficiency and turbid toxin) of CAG. This article explores the pathological mechanism of spleen deficiency and turbid toxin in CAG from the perspectives of iron metabolism, oxidative stress, and lipid peroxidation, providing theoretical support of traditional Chinese medicine for the modern research on CAG. It enriches the modern scientific connotation of the turbid toxicity theory and provides new ideas and breakthrough points for the clinical treatment of CAG.
3.Study on the Pathological Mechanism-Syndrome-Treatment Patterns of Approved Chinese Patent Medicines Targeting Collateral Disorders
Pengli SU ; Peng XU ; Yanhong WANG ; Yaqi ZU ; Run YUAN ; Kun LI ; Yufeng ZHAO
Journal of Traditional Chinese Medicine 2025;66(16):1711-1718
ObjectiveTo explore the pathological mechanism-syndrome-treatment patterns of approved Chinese patent medicines (CPMs) that treat collateral disorders, providing a reference for the principle of "treating different diseases with the same therapy" in collateral pathology. MethodsCPMs that apply treatment strategies based on collateral disorders were identified from the Pharmacodia database by extracting information from the "efficacy" or "indications" sections of drug package inserts. A database was established to extract the names and compositions of the CPMs, as well as their indications, related traditional Chinese medicine (TCM) symptoms, disease locations (affected areas), and pathological factors. Frequency statistics were performed. Using the Apriori algorithm, an association rule analysis was conducted on CPMs and disease-location combinations related to the top three most frequent pathological factor combinations. Core formulas for these combinations were identified and analyzed through drug network analysis and MCODE module clustering. ResultsA total of 660 CPMs targeting collateral disorders were retrieved, involving 299 indications, 323 TCM symptoms, 21 disease locations, 19 pathological factors, and 124 pathological factor combinations. The most frequent pathological factor combinations were blood stasis (involved in 109 CPMs, 16.52%), exogenous wind (外风) -cold-dampness (involved in 43 CPMs, 6.52%), and qi deficiency-blood stasis (involved in 42 CPMs, 6.36%). Analysis of the core formulas for these combinations revealed common ingredients such as Honghua (Carthami Flos), Chuanxiong (Chuanxiong Rhizoma), Danggui (Angelicae Sinensis Radix), and Dilong (Pheretima). ConclusionCollateral disorders involve a wide range of pathogenesis and represent a fundamental mechanism in the onset and development of various diseases, characterized by obstruction and stagnation. The primary therapeutic principle is unblocking of the collaterals. Blood stasis obstructing the collaterals is the core pathological basis, and the strategy of activating blood circulation and resolving stasis to unblock the collaterals should be central to the treatment. The core medication pattern involves combining blood-activating and stasis-resolving herbs with insect-derived medicinals that unblock collaterals. Exogenous wind is often the initiating patholo-gical factor in colla-teral disorders, and the appropriate addition of wind-dispelling herbs can enrich the treatment strategies for such conditions.
4.Expression and clinical significance of zinc finger protein 382 in diffuse large B-cell lymphoma
Sizhe LIU ; Wanhua AN ; Shuli GUO ; Huirui WANG ; Pengli XIAO ; Wanli WANG ; Shuanglin WANG
Journal of Xinxiang Medical College 2024;41(1):37-42
Objective To investigate the expression of zinc finger protein 382(ZNF382)in diffuse large B-cell lymphoma(DLBCL)tissue and its relationship with clinical pathological characteristics and prognosis of DLBCL patients.Methods A total of 57 DLBCL patients admitted to the Department of Hematology,Luoyang Central Hospital from January 2014 to December 2018 were selected as the research subjects.The biopsy pathological specimens and clinical data of DLBCL patients were collected;another 20 patients of reactive proliferative lymph node tissue preserved in the Department of Pathology,Luoyang Central Hospital were taken as the control group.The expression of ZNF382 in DLBCL tissue and reactive proliferative lymph node tissue was detected by En vision two-step method.The difference of ZNF382 expression was compared between DLBCL tissue and reactive proliferative lymph node tissue.The correlations of ZNF382 expression with the clinical features such as age,gender,primary tumor site,Ann Arbor stage,international prognostic index(IPI)score,Hans typing,B-symptoms,bone marrow infiltration,giant masses,Eastern Cooperative Oncology Group(ECOG)score,β2-microglobulin(β2-MG),serum lactate dehydrogenase(LDH),Ki67,and chemotherapy regimen of DLBCL patients were analyzed by univariate analysis;the survival curve was drawed by Kaplan Meier method,and the univariate and multivariate survival analysis were performed by log-rank tests and Cox proportional risk regression models.Results The expression level of ZNF382 in DLBCL tissue was significantly lower than that in reactive proliferative lymph node tissue(Z=-5.056,P<0.01).The expression level of ZNF382 was correlated with IPI score,Ann Arbor stage,Hans typing,B-symptoms,bone marrow infiltration and giant masses of DLBCL patients(P<0.05);the expression level of ZNF382 was not associated to gender,age,primary site,ECOG score,β2-MG,serum LDH,Ki67,and whether the chemotherapy regimen combined with rituximab or not of DLBCL patients(P>0.05).Among the 57 DLBCL patients,the treatment was effective in 36 patients(63.20%)and ineffective in 21 patients(36.80%);the expression level of ZNF382 in tumor tissue of DLBCL patients with effective treatment was significantly higher than that of DLBCL patients with ineffective treatment(Z=-2.895,P<0.05).The 2-year event free survival rate of DLBCL patients in the ZNF382 high expression group was significantly higher than that in the ZNF382 low expression group(x2=17.955,P<0.001).The results of univariate survival analysis showed that female,primary lymph nodes,B-symptoms,bone marrow infiltration,giant masses,IPI score≥3,elevated β2-MG,Ki67>70%,non-germinal center B-cell-like lymphoma,Ann Arbor stageⅢ-Ⅳ and low expression of ZNF382 were risk factors for poor prognosis in DLBCL patients(P<0.05).The results of multivariate analysis showed that primary lymph nodes,Ann Arbor stage Ⅲ-Ⅳ and low expression of ZNF382 were independent influencing factors for poor prognosis in DLBCL patients(P<0.05).Conclusion ZNF382 protein is low expressed in the tumor tissues of DLBCL patients,which is closely related to the occurrence,development and prognosis of DLBCL;and it can be used as an indicator for evaluating the prognosis of DLBCL.
5.Efficacy and safety of branched stent and fenestrated stent for thoracic aortic diseases in short landing zone
Pengli ZHOU ; Yang WANG ; Qinghui ZHANG ; Ling WANG ; Zhengyang WU ; Wenguang ZHANG ; Xinwei HAN
Chinese Journal of Radiology 2024;58(4):422-429
Objective:To compare the efficacy and safety of Castor single-branch stent and in vitro fenestration stent in treating thoracic aortic diseases with insufficient landing zone.Methods:The clinical data of patients with thoracic aortic diseases treated with Castor single-branch stent or in vitro fenestrated stent between December 2017 and June 2021 in the First Affiliated Hospital of Zhengzhou University were retrospectively analyzed. A total of 184 patients were included, 99 patients were treated with Castor branch stent, and 85 patients with in vitro fenestration stent. All patients′ general clinical data, surgical data, perioperative and follow-up clinical and imaging data, and postoperative complications were collected. The χ2 test was used to compare the incidence of complications between the two groups, and the Kaplan-Meier method was used to plot the survival rate without adverse events between the two groups. Results:Stent placement was successful in all patients, and the success rate of the technique was 100%. Other branches were reconstructed in 2 patients in the Castor group and double fenestrated stent were reconstructed in 12 patients in the fenestrated group. The mean operation time of the Castor group was significantly shorter than that of the fenestrated group, the number of patients who received local anesthesia was significantly lower than that of the fenestrated group, and the endoleak rate during follow-up was significantly lower than that of the fenestrated group ( P<0.05). There was no significant difference in the postoperative hospital stay, the incidence rate of perioperative complications, mortality, the incidence rate of neurological complications, new dissection or aneurysm rate, branch stent stenosis rate, second surgical intervention rate, and false lumen thrombosis between the two groups ( P>0.05). The adverse event-free survival rate of the Castor group was slightly higher than that of the fenestrated group, but its difference was not statistically significant ( P>0.05). Conclusion:Castor branch stent and in vitro fenestration stent have good short-term and mid-term efficacy in the treatment of aortic diseases with insufficient landing zone, which are safe and effective options for reconstruction of LSA and other branch arteries.
6.Effect of ADU-S100/doxorubicin in situ vaccine on diffuse large B-cell lymphoma and its mechanism
Pengli XIAO ; Shuli GUO ; Huirui WANG ; Huiyun MAO ; Wanhua AN
Journal of Leukemia & Lymphoma 2024;33(1):29-36
Objective:To explore the antitumor effect of ADU-S100/doxorubicin in situ vaccine on diffuse large B-cell lymphoma (DLBCL) and its mechanism.Methods:The 6-week-old female BALB/c mice were selected, and the bilateral murine subcutaneous B-cell lymphoma model was established with murine B-cell lymphoma A20 cells. The subcutaneous tumor-bearing mice were randomly divided into untreated group (without treatment), ADU-S100 in situ vaccine treatment group (intratumoral injection of interferon gene stimulating factor agonist ADU-S100), doxorubicin in situ vaccine treatment group (intratumoral injection of doxorubicin), and ADU-S100/doxorubicin in situ vaccine treatment group (intratumoral injection of ADU-S100 and doxorubicin) by using random number table method, with 5 mice in each group. The right tumors of the bilateral subcutaneous tumor-bearing mice were defined as proximal tumors, and the left tumors of the bilateral subcutaneous tumor-bearing mice were defined as distal tumors. Only the proximal tumors were treated via the intratumoral route, and the distal tumors were not treated. On day 23 after tumor inoculation, the percentages of CD11c + dendritic cells (DC), CD8 + CD11c + DC and CD80 + CD11c + DC in the spleen of mice in each group were detected by flow cytometry. The splenocytes of mice in each group were stimulated with A20 tumor cell lysate in vitro, the percentages of 5'-ethynyl-2'-deoxyuridine-positive (EdU +) cells and tumor necrosis factor-α-positive (TNF-α +) cells in CD8 + T cells in each in situ vaccine treatment group were detected by flow cytometry, and the killing effect of cytotoxic T lymphocyte (CTL) in each group was measured by using the lactate dehydrogenase (LDH) cytotoxicity assay kit. The mice treated with ADU-S100/doxorubicin in situ vaccine were intraperitoneally injected with anti-mouse CD8α (clone 53-6.7) mAb or isotype control on days 7, 12 and 17 after tumor inoculation to eliminate CD8 + cells. On day 23 after tumor inoculation, the proximal and distal tumor volumes of mice in the ADU-S100/doxorubicin in situ vaccine combined with anti-mouse CD8α (clone 53-6.7) mAb or isotype control treatment group were measured, the percentages of CD8 + T cells and CD8 + CD11c + DC in the spleen of tumor-bearing mice in these two groups were detected by flow cytometry, and the infiltration of CD8 + T cells in the tumor tissues from these two groups was detected by immunohistochemistry (IHC) staining. Results:On days 11, 14, 17, 20 and 23 after tumor inoculation, the proximal and distal tumor volumes of mice in each treated group were lower than those in the untreated group (all P < 0.05). The proportions of CD11c + DC in the spleen of the untreated group, ADU-S100 in situ vaccine treatment group, doxorubicin in situ vaccine treatment group and ADU-S100/doxorubicin in situ vaccine treatment group were (4.92±0.63)%, (7.54±0.84)%, (7.45±0.86)% and (11.63±0.85)%, respectively, and the difference was statistically significant ( F = 72.30, P < 0.001); the proportions of CD8 + CD11c + DC were (1.36±0.34)%, (4.02±0.43)%, (4.22±0.61)% and (6.11±0.73)%, respectively, and the difference was statistically significant ( F = 76.09, P < 0.001); the proportions of CD80 + CD11c + DC were (0.51±0.24)%, (1.69±0.23)%, (1.82±0.25)% and (4.09±0.39)%, respectively, and the difference was statistically significant ( F = 167.40, P < 0.001). The CTL responses and the proportion of EdU + cells and TNF-α + cells in CD8 + T cells in each in situ vaccine treatment group were higher than those in the untreated group (all P < 0.05). Furthermore, the enhanced CTL responses and the increased proportion of EdU + cells and TNF-α + cells in CD8 + T cells were observed in the ADU-S100/doxorubicin in situ vaccine treatment group as compared to the ADU-S100 in situ vaccine treatment group and doxorubicin in situ vaccine treatment group (all P < 0.05). The proportions of CD8 + T cells and CD8 + CD11c + DC in the spleen of mice treated with ADU-S100/doxorubicin in situ vaccine and anti-mouse CD8α mAb were lower than those in ADU-S100/doxorubicin in situ vaccine and isotype control group (both P < 0.05) and both proximal and distal tumor volumes of mice treated with ADU-S100/doxorubicin in situ vaccine and anti-mouse CD8α mAb were larger than those in ADU-S100/doxorubicin in situ vaccine and isotype control group (both P < 0.05). Conclusions:ADU-S100/doxorubicin in situ vaccine can induce profound regression of proximal tumors in bilateral murine subcutaneous B-cell lymphoma model and generate systemic immune responses capable of partially inhibiting distant tumor growth, and the antitumor efficacy of ADU-S100/doxorubicin in situ vaccine may require CD8 + CD11c + DC-mediated CD8 + T cell immune responses.
7.Effect of Coptisine on PI3K/Akt/mTOR Signaling Pathway in Chronic Atrophic Gastritis Rats
Jie WANG ; Pengli DU ; Jiaqi DONG ; Yuewei YANG ; Yunxiao GAO ; Hongyu MA ; Xuemei JIA ; Yuxi GUO ; Bolin LI ; Qian YANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):117-124
ObjectiveTo investigate the therapeutic effect and mechanism of coptisine on chronic atrophic gastritis (CAG) in rats based on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway. MethodA CAG rat model was induced by multiple factors, including sodium salicylate, N-methyl-N′-nitro-N-nitrosoguanidine (MNNG), and irregular feeding. The successfully modeled rats were randomly divided into the model group, folic acid group, and high- and low-dose coptisine groups. The high- and low-dose coptisine groups were given coptisine (50, 10 mg·kg-1, respectively), and the folic acid group was given folic acid at 2 mg·kg-1 for 60 days. The pathological changes were detected by hematoxylin-eosin (HE) staining. The ultrastructure of gastric mucosal cells was observed by electron microscopy. Serum pepsinogen Ⅰ (PGⅠ), pepsinogen Ⅱ (PGⅡ), and PGⅠ/PGⅡ ratio (PGR) were detected by immunoturbidimetry. Serum gastrin-17 (G-17) level was detected by radioimmunoassay. The content of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in serum of rats was detected by enzyme-linked immunosorbent assay (ELSIA). Western blot analysis was used to detect the expression levels of TGF-β1, PI3K, phosphorylated-Akt (p-Akt), mTOR, and phosphatase and tensin homolog deleted on chromosome 10 (PTEN) in gastric mucosa. The mRNA levels of TGF-β1, PI3K, Akt, mTOR, PTEN, microtubule-associated protein light chain 3Ⅱ (LC3Ⅱ), and Beclin-1 were detected by real-time quantitative polymerase chain reaction (Real-time PCR). ResultCompared with the normal group, the model group showed atrophy and reduced number of intrinsic glands in the gastric mucosal tissues, as well as inflammatory cell infiltration. The ultrastructure of gastric mucosal cells in the model group displayed nuclear condensation, reduced and swollen mitochondria, and abnormal structure. The serum levels of G-17, PGⅠ, PGR, and the protein and mRNA levels of PTEN in gastric tissues were significantly lower in the model group (P<0.01), while serum levels of IL-6, IL-1β, TNF-α, and the protein and mRNA levels of TGF-β1, PI3K, Akt, and mTOR in gastric tissues were significantly higher (P<0.01). Compared with the model group, various drug intervention groups showed different degrees of improvement in pathological damage and gastric mucosal cell ultrastructure, significantly increased serum levels of G-17, PGⅠ, and PGR (P<0.05,P<0.01), and significantly decreased levels of IL-6, IL-1β, and TNF-α (P<0.05,P<0.01). The high-dose coptisine group significantly downregulated the protein and mRNA levels of TGF-β1, PI3K, Akt, and mTOR (P<0.05,P<0.01). ConclusionBerberine has a therapeutic effect on CAG in rats, possibly exerting a protective effect on gastric mucosa by inhibiting inflammation and blocking the PI3K/Akt/mTOR signaling pathway.
8.Analysis of Medication Regularity of Prescriptions Containing Cangzhu (Atractylodes Lancea) - Huangbai (Phellodendron Amurense) Based on Data Mining
Pengli SU ; Peng XU ; Yanhong WANG ; Yaqi ZU ; Kun LI ; Yufeng ZHAO
Journal of Traditional Chinese Medicine 2024;65(13):1392-1398
ObjectiveTo analyze medication regularity of presciptions containing Cangzhu (Atractylodes Lancea) - Huangbai (Phellodendron Amurense) based on data mining. MethodsAll the prescriptions containing Cangzhu-Huangbai in Dictionary of Chinese Medical Formulas (《中医方剂大辞典》) and the fifth edition of Chinese Pharmacopoeia (《中华医典》) were retrieved, setting up a database, counting the number of prescriptions involved, the composition of formulas in the prescriptions, the properties of the medicinal, the number of types of diseases and syndromes treated by the prescriptions, and the frequency of their appearances; drawing a network diagram of the medicinal combinations; analysing the medicinal combinations based on the association rules of the Apriori algorithm, and at the same time, analysing the association rules for the medicinal involved in the high-frequency diseases, and the core prescriptions containing Cangzhu-Huangbai pairs for specific diseases were extracted, and the medicinal in the core prescriptions were classified into four categories of raising yang and eliminating dampness, fortifying spleen and eliminating dampness, clearing dampness-heat, and specialised disease-use. ResultsA total of 323 prescriptions were identified, involving 318 Chinese herbal medicines, 52 syndromes, 200 diseases, among which arthralgia, beriberi, atrophy-flaccidity, leukorrhea, hernia-type diseases, headache, and haemorrhoids appeared more than 10 times. Through analyzing the core prescriptions for high-frequency diseases, we had four findings. Firstly, medicinal of spleen-fortifying and dampness-removing were the most frequent used in core prescriptions of Cangzhu-Huangbai pair (353 times), and the medication mode was combination of qi-blood supplement medicinal and qi rectifying medicinal, usually using Huangqi (Astragalus mongholicus), Renshen (Panax ginseng), Processed Gancao (Glycyrrhiza glabra), Baizhu (Atractylodes macrocephala), Danggui (Levisticum officinale), Chenpi (Citrus reticulata), Xiangfu (Cyperus rotundus). Then followed by the combination of heat-clearing and dampness-draining medicinal (213 times), usually using Fangji (Stephania tetrandra), Niuxi (Achyranthes bidentata), and Banxia (Pinellia ternata), and this type of heat-clearing and dampness-draining medicinal is most frequently used for haemorrhoids, and beriberi is the most frequent disease. The combination of yang-raising and dampness-removing medicinal (212 times), often using Qianghuo (Hansenia weberbaueriana), Chaihu (Elephantopus scaber), Fangfeng (Carum carvi), and this type of medicinal is most frequently used for headache, and atrophy-flaccidity is the most frequent disease. The combination of medicinal for specialised diseases was used the least frequently (39 times). ConclusionCangzhu-Huangbai pair were often combined with medicinal of yang-raising and dampness-removing, spleen-fortifying and dampness-removing, and heat-clearing and dampness-draining to expel dampness pathogen, at the same time combined with disease-specific medicinal to treat disease and syndrome together.
9.Association between screen time and psychological behaviors of preschool children in Lanzhou City
Lei HUANG ; Nan NAN ; Yue SU ; Pengli WANG ; Xiaoyan WU ; Jinxian GUO
Chinese Journal of Child Health Care 2024;32(3):242-247
【Objective】 To analyze the associations between different types of video screen time and psychological behaviors of preschool children, in order to provide evidence for promoting the development of children′s mental health. 【Methods】 From February to March 2023, a total of 1 361 parents of children aged 3 - 6 years from 6 kindergartens of Lanzhou were surveyed by cluster sampling method.Parents were surveyed to obtain information about the video use, and the children′s Strengths and Difficulties questionnaire (parent version) was used to assess children′s psychological and behavioral problems. 【Results】 The rate of daily screen time exceeding standard was 36.96% (503/1 361).The screen time was mainly spent in watching TV cartoons, followed by educational APP.The detection rate of abnormal total difficulty score was 11.61% (158/1 361), and the abnormalities of peer communication (32.26%) and prosocial behavior (12.34%) were the most prominent.After adjusting for related factors by multiple Logistic regression analysis, total screen time≥2h/d (OR=1.802) was found to be a risk factor for abnormal total difficulty score; watching TV cartoons≥2h/d was a risk factor for abnormal total difficulty score (OR=2.409) and peer communication (OR=2.222); playing games≥1h/d was a risk factor for abnormal total difficulty score, emotional symptoms, conduct problems, hyperactive behavior, and abnormalities of peer communication, the differences were all statistically significant (P<0.05).However, educational APP screen time<1h/d was a protective factor for abnormal total difficulty score(OR=0.615) and prosocial behavior (OR=0.549), but educational APP screen time≥2h/d was a risk factor for conduct problems (OR=2.302), the differences were all statistically significant (P<0.05). 【Conclusions】 The screen time of preschool children in Lanzhou cannot be ignored, and there is a significant correlation between overuse and children′s psychological and behavioral problems.Parents and schools should attach importance to the parent-child and peer interaction of preschool children and strengthen the intervention of preschool children′s video behavior.
10.Antitumor immune response of stimulator of interferon genes-based Dickkopf-related protein 1-targeted vaccine in multiple myeloma
Pengli XIAO ; Shuli GUO ; Huirui WANG ; Huiyun MAO ; Wanhua AN
Journal of Xinxiang Medical College 2024;41(10):911-918
Objective To explore whether stimulator of interferon genes(STING)agonist ADU-S100 could enhance the antitumor immune response of a chitosan(CS)nanoparticle-mediated DNA vaccine containing a tumor-specific antigen Dickkopf-related protein 1(DKK1)in multiple myeloma(MM).Methods CS-DNA nanoparticles were prepared by using the compound coprecipitation method.The particle sizes and Zeta potential of the CS-DNA nanoparticles were measured by using the Zetasizer Nano-ZS laser particle size analyzer.The DNA protection effect and in vivo DNA expression efficiency of the CS-DNA nanoparticles were assessed by using gel retardation assay and Western blot,respectively.The lentiviruses expressing human DKK1(hDKK1)genes were used to establish MPC-11 cells(MPC-11-hDKK1)which stably expressed hDKK1,and the MPC-11-hDKK1 cells were subcutaneously given to mice to construct tumor models.The tumor-bearing mice were randomly divided into a control group(intramuscular injection of CS-pcDNA3.1),an ADU-S100 immunization group(subcutaneous injection of ADU-S100),a CS-pDKK1 immunization group(intramuscular injection of CS-pDKK1)and an ADU-S1OO/CS-pDKK1 co-immunization group(intramuscular injection of CS-pDKK1+subcutaneous injection of ADU-S100),with 5 mice in each group.The tumor-bearing mice in each group were immunized 3 times at 10-day intervals according to the corresponding immunization schedule.The size of tumor was measured every week.On day 42 after MPC-11-hDKK1 cell inoculation,the tumor weight of mice in each immunization group was measured;the percentages of CD11c+dendritic cell(DC),CD8+CD11c+DC and major histocompatibility complex class Ⅱ(MHCII)+CD11c+DC subsets in the spleen of mice in each immunization group were detected by using flow cytometry.The splenocytes of mice in each group were stimulated with recombinant hDKK-1 protein in vitro,the percentage of EdU+cells in CD8+T lymphocytes in each immunization group was detected by using flow cytometry,and the killing effect of cytotoxic T lymphocyte(CTL)in each group was assessed by using the lactate dehydrogenase(LDH)cytotoxicity assay kit.Results The particle size and Zeta potential of the CS-DNA nanoparticles were(204.3±2.31)nm and(15.47±1.01)mV,respectively.Gel retardation assay showed that DNA enveloped in CS nanoparticles could be completely retarded.Western blot analysis indicated that CS-DNA nanoparticles could be effectively expressed in vivo.The relative expression of DKK1 protein was significantly higher in MPC-11-hDKK1 cells than in MPC-11-Ctrl cells(P<0.05).On days 7 and 14 after MPC-11-hDKK1 cell inoculation,there was no significant difference in tumor volume of mice between the ADU-S100 immunization group,CS-pDKK1 immunization group,ADU-S100/CS-pDKK1 co-immunization group and the control group(P>0.05);on days 21,28,35 and 42 after MPC-11-hDKK1 cell inoculation,the tumor volumes of mice in the ADU-S100 immunization group,CS-pDKK1 immunization group and ADU-S100/CS-pDKK1 co-immunization group were significantly lower than those in the control group(P<0.05);the tumor volume of mice in the ADU-S100/CS-pDKK1 co-immunization group was significantly lower than that in the ADU-S100 immunization group and CS-pDKK1 immunization group(P<0.05).On day 42 after MPC-11-hDKK1 cell inoculation,the tumor weight of mice in the ADU-S100 immunization group,CS-pDKK1 immunization group and ADU-S1 OO/CS-pDKK1 co-immunization group was significantly lower than that in the control group(P<0.05);the tumor weight of mice in the ADU-S100/CS-pDKK1 co-immunization group was significantly lower than that in the ADU-S100 immunization group and CS-pDKK1 immunization group(P<0.05).The proportions of CD11c+DC,CD8+CD11c+DC and MHCII+CD11c+DC subsets in the spleen of mice in the ADU-S100 immunization group,CS-pDKK1 immunization group and ADU-S100/CS-pDKK1 co-immunization group were significantly higher than those in the control group(P<0.05).The proportions of CD11c+DC,CD8+CD11c+DC and MHCII+CD11c+DC subsets in the spleen of mice in the ADU-S100/CS-pDKK1 co-immunization group were significantly higher than those in the ADU-S100 immunization group and CS-pDKK1 immunization group(P<0.05).The CTL killing effect and the proportion of EdU+cells in CD8+T lymphocytes in the ADU-S100 immunization group,CS-pDKK1 immunization group and ADU-S1OO/CS-pDKK1 co-immunization group were significantly higher than those in the control group(P<0.05);the CTL killing effect and the proportion of EdU+cells in CD8+T lymphocytes in the ADU-S100/CS-pDKK1 co-immunization group were significantly higher than those in the ADU-S100 immunization group and CS-pDKK1 immunization group(P<0.05).Conclusion STING agonist ADU-S100 can significantly improve the antitumor immunity of the CS-pDKK1 nanoparticle vaccine in MM,and this vaccine strategy provides a potential treatment approach for MM.

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