1.Inhibition of Epithelial-mesenchymal Transition Mechanism in Chronic Atrophic Gastritis Rats by Banxia Xiexintang via Regulating IL-17/ERK/C/EBPβ Signaling Pathway
Wenyu WU ; Xinyu ZENG ; Hao LI ; Weiqi SUN ; Jiahui REN ; Yang YU ; Tingting ZHOU ; Aili XU ; Wei WEI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):1-10
ObjectiveThis study aimed to investigate the action mechanism by which Banxia Xiexintang (BXT) inhibits epithelial-mesenchymal transition (EMT) in chronic atrophic gastritis (CAG) rats by regulating the interleukin-17(IL-17)/extracellular regulated protein kinases(ERK)/CCAAT enhancer binding protein β(C/EBPβ)signaling pathway, thereby providing new theoretical evidence for the treatment of CAG with classic traditional Chinese medicine formulas. MethodsA CAG rat model was established by using the combined factor method. After successful modeling, the rats were randomly divided into the model group, low-, medium-, and high-dose groups (0.549, 1.098, 2.196 g·kg-1, respectively) of BXT, and the positive drug group (vitacoenzyme, 0.3 g·kg-1). A normal control group was also set up. After 8 weeks of intervention, the pathological changes of gastric tissue were evaluated. The enzyme-linked immunosorbent assay (ELISA) was used to detect the contents of IL-17, tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (COX-2), and C/EBPβ in serum, as well as the contents of EMT markers in gastric mucosal tissue including E-cadherin, N-cadherin, and vimentin. The immunohistochemistry method was employed to determine the localization and protein expression levels of IL-17, p-ERK, and C/EBPβ in gastric mucosal tissue. Western blot was used to detect the protein expressions of C/EBPβ, ERK, and its phosphorylated form (p)-ERK in gastric mucosa. Real-time polymerase chain reaction (Real-time PCR) was applied to measure the mRNA expression levels of ERK, COX-2, and C/EBPβ in gastric mucosa. ResultsCompared with those in the normal control group, the rats in the model group showed gastric mucosal glandular atrophy and inflammatory cell infiltration. The protein and their related mRNA expressions of C/EBPβ, ERK, and p-ERK in gastric mucosa were significantly increased (P<0.05,P<0.01). The levels of IL-17, TNF-α, COX-2, and C/EBPβ in serum were significantly increased (P<0.01). The contents of N-cadherin and vimentin in gastric mucosal tissue were significantly increased, while the content of E-cadherin was significantly decreased (P<0.01). Compared with the model group, after intervention with different doses of BXT, the pathological damage of the gastric mucosa was improved to varying degrees. The protein and mRNA expressions of C/EBPβ, ERK, and p-ERK in gastric mucosa were significantly reduced (P<0.05,P<0.01). The levels of IL-17, TNF-α, COX-2, and C/EBP β in serum were significantly decreased (P<0.01). The contents of N-cadherin and vimentin in gastric mucosa tissue were decreased, while the content of E-cadherin was increased (P<0.05,P<0.01). ConclusionBXT can effectively improve the pathological damage of gastric mucosal tissue in CAG rats. Its action mechanism may be related to reducing the levels of IL-17 and TNF-α in serum, regulating the IL-17/ERK/C/EBPβ signaling pathway and inhibiting the EMT process.
2.Current Research Status,Challenges,Differentiation and Treatment Strategies of Traditional Chinese Medicine for Gastroesophageal Reflux Disease
Fengyun WANG ; Mi LYU ; Bingduo ZHOU ; Beihua ZHANG ; Yi WANG ; Tingting XU ; Cong HE ; Xiaokang WANG ; Xin LIU ; Yang WANG ; Kaiyue HUANG ; Lusi XU ; Xudong TANG
Journal of Traditional Chinese Medicine 2026;67(4):392-396
This article systematically reviews the current research status as well as diagnosis and treatment strategies of traditional Chinese medicine (TCM) for gastroesophageal reflux disease (GERD). Studies demonstrate that TCM, based on the "disease-syndrome combination" approach, exhibits multi-target advantages in alleviating symptoms of various GERD subtypes, promoting mucosal repair, regulating emotions, and facilitating the reduction of western medication. To address clinical challenges such as symptom overlap and limited therapeutic efficacy, strategies have been proposed including "treating different diseases with the same method" and integrated regulation based on viscera correlation. Future efforts should focus on elucidating the mechanisms of compound prescriptions, promoting TCM drug development under the "three-combination" evaluation framework that integrates TCM theory, human experience and clinical trial evidence, and optimizing integrated traditional and western medicine models to enhance GERD management.
3.Mechanism of Yangjing Zhongyutang in Regulating SIRT1/PGC-1α Signaling Pathway to Promote Mitochondrial Function and Alleviate Oxidative Stress Damage in Rats with Diminished Ovarian Reserve
Ping ZHANG ; Lijuan YANG ; Shenghui CHEN ; Wenliang YAO ; Yuliang ZHOU ; Ling MA ; Huiying WU ; Yanwen XU ; Ziyan ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):46-55
ObjectiveTo observe the effects of Yangjing Zhongyutang (YJZYT) on mitochondrial biogenesis and oxidative stress damage mediated by the silent information regulator 1 (SIRT1)/peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1α) signaling pathway in cyclophosphamide (CTX)-induced rats with diminished ovarian reserve (DOR), and to explore its mechanism in improving ovarian reserve function and follicular development. MethodsForty-two 8-week-old female SD rats with normal estrous cycles were randomly divided into a blank control group (n=7) and a model group (n=35). Rats in the model group received a single intraperitoneal injection of CTX (90 mg·kg-1) to establish the DOR model. After modeling, estrous cycles were monitored for 7 consecutive days, and model success was confirmed based on criteria for estrous cycle disruption. After successful modeling, rats were divided into groups for intervention: estradiol valerate group (0.09 mg·kg-1), and YJZYT high-, medium-, and low-dose groups (19.98, 9.99, 5.00 g·kg-1). The blank control group and model group were given an equal volume of distilled water by gavage. All groups received daily gavage once for 4 consecutive weeks. The general state, body weight, and ovarian wet weight of rats were observed and recorded, and the ovarian organ index was calculated. Enzyme-linked immunosorbent assay (ELISA) was used to measure serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), anti-Müllerian hormone (AMH), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px). Hematoxylin-eosin (HE) staining was performed to observe ovarian histomorphological changes and follicular development status. Immunofluorescence was used to detect reactive oxygen species (ROS) expression levels. Colorimetric assays were employed to measure adenosine triphosphate (ATP) and malondialdehyde (MDA) content in ovarian tissues. Quantitative Real-time polymerase chain reaction (Real-time PCR) was used to detect mitochondrial DNA (mtDNA) copy number and the mRNA expression levels of key genes including SIRT1, PGC-1α, nuclear respiratory factor 1 (NRF1), and mitochondrial transcription factor A (TFAM). Western blot was performed to detect the protein expression levels of SIRT1, PGC-1α, NRF1, and TFAM. ResultsCompared with the blank group, rats in the model group exhibited disrupted estrous cycles, obviously reduced body weight, and decreased ovarian index (P<0.05). Ovarian histopathology revealed cortical thinning, loose structure, and a significant reduction in both primordial and growing follicles (P<0.01). Serum FSH and LH levels were significantly elevated (P<0.01), while E2 and AMH levels were obviously reduced (P<0.05, P<0.01). ATP content and mtDNA copy number decreased in ovarian tissue (P<0.01), ROS expression increased, MDA levels rose, while SOD and GSH-Px activities obviously decreased (P<0.05, P<0.01), mRNA and protein expression levels of SIRT1, PGC-1α, NRF1, and TFAM were obviously downregulated (P<0.05, P<0.01). After treatment, compared with the model group, body weight and ovarian index obviously recovered in rats administered various doses of YJZYT (P<0.05), serum E2 and AMH levels increased, while FSH and LH levels obviously decreased (P<0.05, P<0.01), ovarian tissue ATP content and mtDNA copy number were up-regulated, ROS and MDA levels decreased, and antioxidant enzymes SOD and GSH-Px activity obviously increased (P<0.05, P<0.01), Gene and protein expression levels related to the SIRT1/PGC-1α /NRF1/TFAM signaling pathway were obviously up-regulated compared to the model group (P<0.05, P<0.01), HE staining revealed that ovarian structure gradually recovered to integrity in all treatment groups, with a obviously increase in the number of primordial and growing follicles (P<0.05, P<0.01). Granulosa cells were neatly arranged, indicating marked improvement in ovarian function. ConclusionYJZYT may improve ovarian function and follicular development in rats with diminished ovarian reserve by activating the SIRT1/PGC-1α signaling pathway, promoting mitochondrial biogenesis, enhancing mitochondrial function, and alleviating oxidative stress damage.
4.A meta-analysis of risk factors for residual back pain after vertebral augmentation for osteoporotic vertebral compression fractures
Peng YANG ; Chenghan XU ; Yingjie ZHOU ; Xubin CHAI ; Hanjie ZHUO ; Lin LI ; Jinyu SHI
Chinese Journal of Tissue Engineering Research 2026;30(3):731-739
OBJECTIVE:Patients with osteoporotic vertebral compression fractures still have residual back pain after vertebral augmentation.The current research is characterized by limited sample size,complex confounding factors,and inconsistent research results.To gain a deeper understanding of this phenomenon,the aim of this study was to identify and evaluate the risk factors for residual back pain after surgery through a systematic review and meta-analysis.METHODS:A comprehensive search was conducted in CNKI,VIP,WanFang,CBMdisc,PubMed,The Cochrane Library,Embase,and Web of Science for case-control studies on residual back pain after vertebral body augmentation for osteoporotic vertebral compression fractures from database inception to July 2024.The search terms were a combination of subject terms and free terms.The basic information,patient characteristics,surgical-related indicators,and risk factors for surgical back pain of the included studies were extracted.After evaluating the bias risk of all included studies,a meta-analysis was conducted using Stata 14.0 software on the relevant indicators.RESULTS:(1)21 case-control studies with a total of 8 043 patients were included.Among them,965 patients developed back pain.The quality score of all 21 studies was ≥7.(2)The meta-analysis results showed that age(WMD=0.98,95%CI:0.40-1.56,P=0.010),bone mineral density(WMD=-0.28,95%CI:-0.34 to-0.21,P=0.000),the number of vertebral fractures(OR=3.50,95%CI:2.65-4.62,P=0.000),thoracolumbar fracture index(OR=3.65,95%CI:2.61-5.11,P=0.000),cement volume(OR=6.89,95%CI:2.62-18.17,P=0.000),and cement distribution(OR=2.38,95%CI:1.93-2.93,P=0.000)were risk factors for the development of back pain after vertebral body augmentation in patients with osteoporotic vertebral compression fractures.CONCLUSION:Current evidence indicates that age,bone mineral density,the number of vertebral fractures,thoracolumbar fracture index,bone cement injection volume,and the distribution of bone cement are risk factors for low back pain.Specifically,bone mineral density,the number of vertebral fractures,thoracolumbar fracture index,and non-uniform distribution of bone cement are identified as independent risk factors for low back pain.Patients exhibiting these high-risk factors require vigilant monitoring and prompt intervention to mitigate the occurrence of clinical low back pain,thereby enhancing patient outcomes and quality of life.
5.Comparison of sleep EEG power spectral density between depressive episode patients and schizophrenia patients with suicidal behavior
Jingwen LIU ; Yunfei ZHOU ; Jingchu HU ; Jiaoyan ZHOU ; Junwei YANG ; Jie LIANG ; Hong XU ; Yu CANG ; Shimeng MA
Sichuan Mental Health 2026;39(1):50-57
BackgroundPatients with depressive episode and schizophrenia have a high risk of suicide. The sleep electroencephalogram power spectral density characteristics of patients with depressive episode accompanied by suicidal behavior and those with schizophrenia may be different, but there is currently a lack of direct comparative studies on these two groups of patients. ObjectiveTo compare the sleep electroencephalogram power spectral density between depressive episode and schizophrenic patients with suicidal behavior, in order to provide references for exploring predictive indicators of suicidal behavior. MethodsFrom June 2018 to December 2020, 20 patients with depressive episode and 20 patients with schizophrenia who had committed suicide within the past month and were treated at the outpatient department of Shenzhen Kangning Hospital were selected. All of them met the diagnostic criteria for depressive episode or schizophrenia as defined in the International Classification of Diseases, tenth edition (ICD-10). Using a random sampling method, 20 volunteers with matching gender and age to the patient groups were selected from the Cuiping community in Shenzhen as the control group. The subjective sleep of the patients was evaluated using the Insomnia Severity Index (ISI), the Dysfunctional Belief and Attitude about Sleep (DBAS), the Disturbing Dreams and Nightmare Severity Index (DDNSI), and the Epworth Somnolence Scale (ESS). The objective sleep of the patients was assessed using polysomnography. The sleep electroencephalogram was filtered and the power spectral density of the brain wave was analyzed and processed for all the subjects. The subjective and objective sleep conditions of the two patient groups were compared, and the sleep electroencephalogram power spectral density of the patient groups and the control group were also compared. ResultsA comparison of subjective and objective sleep conditions between patients with depressive episode accompanied by suicidal behavior and patients with schizophrenia accompanied by suicidal behavior showed no statistically significant differences (P>0.05). Comparisons of sleep electroencephalogram power spectral density in the W stage (average power of α wave, total power of δ wave, average power of δ wave, average power of θ wave), N1 stage (average power of β wave, total power of α wave, total power of δ wave), N2 stage (total power of α wave, average power of α wave, total power of δ wave, average power of δ wave), N3 stage (average power of α wave, average power of δ wave), and R stage (total power of α wave, average power of α wave, total power of δ wave, average power of δ wave) between patients with depressive episode accompanied by suicidal behavior, patients with schizophrenia accompanied by suicidal behavior, and the control group showed statistically significant differences (P<0.05 or 0.01). The total power of δ wave in the W stage and the average power of β wave and δ wave in the N1 stage were higher in two patient groups were higher than those of the control group. The total power of α wave and the average power of α wave in the N2 stage were lower than those of the control group, while the average power of δ wave was higher than that of the control group. The average power of α wave in the N3 stage of both patient groups were lower than that of the control group, while the average power of δ wave was higher than that of the control group. The total power and average power of α wave in the R stage were lower than those of the control group, while the total power and average power of δ wave were higher than those of the control group. All the differences were statistically significant. Patients with depressive episode accompanied by suicidal behavior had higher average powers of α wave, δ wave, and θ wave in the W stage compared with the control group, while the total power of α wave in the N1 stage was lower in the former group. All these differences were statistically significant (P<0.05). ConclusionThe depressive episode patients accompanied by suicidal behavior have highly overlapping sleep electroencephalogram abnormal patterns with those of schizophrenia patients, mainly manifested as a general decrease in α wave power (N2, N3, R stage) and a general increase in δ wave power (W, N1, N2, N3, R stage) as well as β wave power in N1 stage. At the same time, patients with depressive episode accompanied by suicidal behavior also show specific changes, including an increase in the average power of α and θ waves during the wakefulness period (W stage), and a decrease in the total power of α wave in N1 stage. [Funded by Guangdong Province High-level Clinical Key Specialty (with supporting funds from Shenzhen City) (number, SZGSP013); Shenzhen Key Medical Discipline (number, SZXK041); Shenzhen Clinical Medicine Research Center Project (number, 20210617155253001)]
6.Herbal Textual Research on Bambusae Succus in Famous Classical Formulas
Yu SHI ; Feng ZHOU ; Yihan WANG ; Yanmeng LIU ; Ming YANG ; Zhiping CHEN ; Jiangshan ZHANG ; Conglong XU ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(9):231-239
This article systematically reviews and examines the historical evolution of Bambusae Succus as a medicinal material, covering aspects such as nomenclature, origin, geographical distribution, harvesting and processing methods, quality assessment, therapeutic effects and indications, by consulting ancient herbal texts, medical compendia, and modern literature. The aim is to provide a reference for the development and utilization of famous classical formulas containing this herb. Research indicated that Bambusae Succus was first documented in the Shennong Bencaojing during the Han dynasty, with Zhuli being the standard name used throughout history, alongside aliases like Zhuzhi, Zhuyou and Huoquan. Historically, the primary source of Bambusae Succus has been Phyllostachys nigra var. henonis(Danzhu), although other species such as Pleioblastus amarus and Bambusa emeiensis have also been used medicinally. Ancient records predominantly noted its origin in Yizhou(present-day Chengdu and surrounding areas in Sichuan) and the Wuling region(between present-day Hunan, Guangdong, Guangxi and Jiangxi provinces), while contemporary sources are mainly from regions south of the Yangtze River and southwestern China. Traditionally, Bambusae Succus was harvested from bamboo that had grown for exactly one year, today, it can be collected year-round without strict age requirements. Ancient preparation methods included direct fire roasting or dry distillation, whereas modern industrial production employs dry distillation, reflux extraction, and percolation. In terms of quality evaluation, ancient texts considered a sweet taste to be superior, while today, clarity and transparency are prioritized. Historically, Bambusae Succus was characterized as sweet and cold nature, targeting the lung and stomach meridians, with uses evolving from clearing heat and resolving phlegm to nourishing Yin, moistening dryness, and relaxing tendons and unblocking meridians. Modern descriptions classify it as sweet, bitter, and cold in nature, affecting the heart, liver, and lung meridians, with functions including clearing heat, resolving phlegm, and facilitating orifices. It is indicated for conditions such as stroke with phlegm confusion, lung heat with phlegm congestion, convulsions, epilepsy, excessive phlegm in febrile diseases, high fever with thirst, irritability during pregnancy, and tetanus, with more clearly defined applications. Based on the results of the research, it is recommended that when developing and utilizing famous classical formulas containing Bambusae Succus, the one-year-old Phyllostachys nigra var. Henonis, which has been highly praised throughout history, should be selected as the source material. Industrial production should adopt the dry distillation method. Furthermore, in-depth research should be conducted on the modern technological characterization of the traditional quality control indicator of sweet taste, and reasonable modern quality control standards should be established.
7.CEACAM6 Expression is Associated with Immune Infiltration and Poor Prognosis in Esophageal Squamous Cell Carcinoma
Jiahui LI ; Enwei XU ; Wei CUI ; Yuanyuan ZHAO ; Keqing KANG ; Peng BU ; Guohai ZHAO ; Yang ZHOU
Cancer Research on Prevention and Treatment 2026;53(3):194-202
Objective To investigate the expression of carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) in esophageal squamous cell carcinoma (ESCC) and analyze its correlation with immune cell infiltration and patient prognosis. Methods Three ESCC datasets (GSE161533, GSE26886, and GSE23400) from the GEO database were analyzed to identify differentially expressed genes. CEACAM6 was identified as a key gene through survival analysis. Its expression, prognostic value, and relationship with immune cell infiltration were further explored using databases, such as TIMER. Tissue samples were collected from 162 patients with ESCC. Immunohistochemistry was performed to detect the expression of CEACAM6, immune cell markers (CD4, CD8, CD20, and CD56), and immune checkpoint molecules (HHLA2 and CD40LG). Correlations between CEACAM6 expression and clinicopathological features, immune cell infiltration, and immune checkpoints were analyzed. Results Bioinformatic analysis and clinical sample validation confirmed that CEACAM6 expression was significantly upregulated in ESCC tissues compared with adjacent nontumor tissues (P<0.05). High CEACAM6 expression was closely associated with advanced clinical stage (AJCC Ⅲ-Ⅳ), high T stage (T3-T4), lymph node metastasis, nonulcerative type, and poor prognosis. Furthermore, CEACAM6 expression levels were positively correlated with the infiltration density of CD8+ T cells, CD4+ T cells, and CD20+ B cells within the tumor microenvironment and with the expression of the immune checkpoint molecules HHLA2 and CD40LG (all P<0.05). Conclusion CEACAM6 serves as an independent poor prognostic factor for ESCC. Its high expression is implicated in the modulation of the tumor immune microenvironment by correlating with specific immune cell infiltration and immune checkpoint molecules, suggesting its potential as a novel prognostic biomarker and immunotherapeutic target for ESCC.
8.A meta-analysis of risk factors for residual back pain after vertebral augmentation for osteoporotic vertebral compression fractures
Peng YANG ; Chenghan XU ; Yingjie ZHOU ; Xubin CHAI ; Hanjie ZHUO ; Lin LI ; Jinyu SHI
Chinese Journal of Tissue Engineering Research 2026;30(3):731-739
OBJECTIVE:Patients with osteoporotic vertebral compression fractures still have residual back pain after vertebral augmentation.The current research is characterized by limited sample size,complex confounding factors,and inconsistent research results.To gain a deeper understanding of this phenomenon,the aim of this study was to identify and evaluate the risk factors for residual back pain after surgery through a systematic review and meta-analysis.METHODS:A comprehensive search was conducted in CNKI,VIP,WanFang,CBMdisc,PubMed,The Cochrane Library,Embase,and Web of Science for case-control studies on residual back pain after vertebral body augmentation for osteoporotic vertebral compression fractures from database inception to July 2024.The search terms were a combination of subject terms and free terms.The basic information,patient characteristics,surgical-related indicators,and risk factors for surgical back pain of the included studies were extracted.After evaluating the bias risk of all included studies,a meta-analysis was conducted using Stata 14.0 software on the relevant indicators.RESULTS:(1)21 case-control studies with a total of 8 043 patients were included.Among them,965 patients developed back pain.The quality score of all 21 studies was ≥7.(2)The meta-analysis results showed that age(WMD=0.98,95%CI:0.40-1.56,P=0.010),bone mineral density(WMD=-0.28,95%CI:-0.34 to-0.21,P=0.000),the number of vertebral fractures(OR=3.50,95%CI:2.65-4.62,P=0.000),thoracolumbar fracture index(OR=3.65,95%CI:2.61-5.11,P=0.000),cement volume(OR=6.89,95%CI:2.62-18.17,P=0.000),and cement distribution(OR=2.38,95%CI:1.93-2.93,P=0.000)were risk factors for the development of back pain after vertebral body augmentation in patients with osteoporotic vertebral compression fractures.CONCLUSION:Current evidence indicates that age,bone mineral density,the number of vertebral fractures,thoracolumbar fracture index,bone cement injection volume,and the distribution of bone cement are risk factors for low back pain.Specifically,bone mineral density,the number of vertebral fractures,thoracolumbar fracture index,and non-uniform distribution of bone cement are identified as independent risk factors for low back pain.Patients exhibiting these high-risk factors require vigilant monitoring and prompt intervention to mitigate the occurrence of clinical low back pain,thereby enhancing patient outcomes and quality of life.
9.Professor WU Rongzu's Clinical Experience in Treating Irritable Bowel Syndrome with Constipation Using Fuyang Tongmi Decoction (扶阳通秘汤)
Yihao YANG ; Junran ZHU ; Wendi WU ; Liyun JIANG ; Yueqiu DONG ; Yueqing CAI ; Ruibin ZHOU ; Yunjiao XU ;
Journal of Traditional Chinese Medicine 2026;67(10):1049-1051
This paper introduces professor WU Rongzu's clinical experience in using Fuyang Tongmi Decoction (扶阳通秘汤, FTD) to treat irritable bowel syndrome with constipation (IBS-C). It is believed that yang qi depletion, water cold, earth dampness, and wood constraint are the key pathogenesis.The treatment principle is warming water, drying the earth and venting wood, with the basic formula FTD adjusted according to the symptoms. This approach aims to transport the qi movement of the middle jiao (焦) and support the recovery of intestinal function of directing turbidity downward, providing a treatment strategy for IBS-C caused by yang deficiency.
10.Pathological changes and macrophage polarization in the liver and spleen of mice infected with Angiostrongylus cantonensis
Xiaoyu QIN ; Yuchun CAI ; Yang HONG ; Fanna WEI ; Yahong HU ; Yumeng CAI ; Yuan HU ; Ting ZHANG ; Xiaojin MO ; Bin XU ; Yan LU ; Jiahui SUN ; Yan ZHOU ; Zelin ZHU ; Muxin CHEN
Chinese Journal of Schistosomiasis Control 2026;38(2):169-183
Objective To investigate the temporal changes in pathological damage and macrophage polarization in liver and spleen tissues of mice infected with Angiostrongylus cantonensis, and to preliminarily unravel the peripheral immune responses during the early stage of A. cantonensis infection. Methods Forty female BALB/c mice at ages of 6 to 8 weeks were randomly divided into four groups, including the control group and 7-, 14-, and 21-day infection groups, with 10 mice in each group. Each mouse in the infection groups was inoculated with 30 third-stage (L3) larvae of A. cantonensis by oral gavage, and five mice were randomly selected from each infection group on days 7, 14, and 21 post-infection, while mice in the control group were given the same volume of physiological saline and five mice were randomly selected from the control group on the day of oral gavage. Mouse liver and spleen tissues were sampled. The histopathological changes of mouse liver and spleen tissues were observed using hematoxylin and eosin (HE) staining, and the percentage of positive staining area and the co-localization positive rates of the macrophage surface antigens F4/80, CD86, and CD206 were quantified in mouse liver and spleen tissues using immunohistochemical and immunofluorescence staining. In addition, five mice were collected from each infection group on days 7, 14, and 21 post-infection, and five mice were collected from the control group on the day of oral gavage. Mouse liver and spleen tissues were sampled for detection of macrophage markers CD86 and CD206 and macrophage phenotyping using flow cytometry, and the expression of M1 macrophage markers, including inducible nitric oxide synthase (Nos2), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and M2 markers, including arginase 1 (Arg1), mannose receptor C-type 1 (Mrc1) and chitinase-like protein 3 (Chil3) was quantified in mouse liver and spleen tissues using real-time quantitative PCR (RT-qPCR) assay. Results Proliferative lesions of the hepatocyte were observed in mouse liver tissues and the follicular structures of the mouse spleen white pulp were disrupted 21 days post-infection with A. cantonensis. Immunohistochemical staining showed that there were significant differences in the percentages of F4/80, CD86 and CD206 positive staining areas in the liver and spleen tissues among the four groups of mice (F = 242.40, 197.14, 183.19, 157.65, 242.35 and 146.24; all P values < 0.001), and the percentages of positive staining in the liver and spleen tissues of mice in the 14-day infection group [(4.45 ± 0.51)%, (3.74 ± 0.67)%, (8.32 ± 0.72)%, (16.56 ± 1.14)%, (11.62 ± 0.52)%, and (8.29 ± 0.72)%, respectively] and the 21-day infection group [(3.70 ± 0.11)%, (3.22 ± 0.43)%, (11.53 ± 1.03)%, (12.59 ± 1.05)%, (9.02 ± 0.83)%, and (11.67 ± 1.10)%, respectively] were higher than in the control group [(0.35 ± 0.16)%, (0.40 ± 0.02)%, (0.93 ± 0.05)%, (2.78 ± 0.26)%, (2.33 ± 0.20)%, and (1.85 ± 0.20)%, respectively] (all P values < 0.05). Immunofluorescence staining showed significant differences in the positive rates of F4/80 co-localization with CD86 and CD206 in mouse liver and spleen tissues among the four groups (F = 24.42, 25.28, 54.51 and 130.55; all P values < 0.001). Flow cytometry detected significant differences in the proportions of CD86+ and CD206+ macrophages in mouse liver and spleen tissues among the four groups (F = 67.98, 18.41, 29.77, 172.80; all P values < 0.001), and the proportions of CD206+ macrophages in the liver and spleen of the 21-day infection group were significantly higher than those in the control group [(9.25 ± 2.55)% vs (3.83 ± 0.72)%, and (4.22 ± 0.56)% vs (0.47 ± 0.18)%, respectively] (both P values < 0.05). In addition, RT-qPCR assay quantified significant differences in the relative mRNA expression of M1 macrophage markers (IL-1β, TNF-α and Nos2) and M2 macrophage markers (Arg1, Chil3 and Mrc1) in mouse liver and spleen tissues among the four groups (F = 41.30, 31.82, 199.33, 19.96, 62.01, 119.76, 23.67, 95.90, 72.27, 82.59, 123.41 and 29.75; all P values < 0.05). Conclusions A. cantonensis infection may cause progressive pathological damage in mouse liver and spleen tissues, accompanied by dynamic temporal changes in macrophage polarization. M1 macrophage polarization predominates at the early stage of A. cantonensis infection and shifts towards M2 polarization at the later stages, suggesting that M2 polarization may participate in immune regulation at late stages of A. cantonensis infection by suppressing excessive inflammatory responses and promoting tissue repair.

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