1.Molecular Crosstalk Mechanisms of Shoutai Wan and Juyuan Jian on Maternal-fetal Interface Subcellular Clusters in CBA/J×DBA/2 Recurrent Pregnancy Loss Model
Jingxin GAO ; Qiuping CHEN ; Xiaoyan ZHENG ; Pengfei ZENG ; Rui ZHOU ; Yancai TANG ; Qian ZENG ; Wenli GUO ; Jinzhu HUANG ; Weijun DING ; Linwen DENG ; Hang ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):70-87
ObjectiveTo systematically compare the differential regulation of the maternal-fetal interface cell lineages and communication networks in the CBA/J×DBA/2 mouse model of recurrent pregnancy loss (RPL) by the two classic therapeutic methods-tonifying the kidney to stabilize the fetus and invigorating the spleen to stabilize the fetus (Shoutai Wan, Juyuan Jian)-of traditional Chinese medicine (TCM) at the single-cell resolution and clarify their modern scientific connotations. MethodsFemale non-pregnant CBA/J mice were caged with male BALB/c (blank group) and DBA/2 (modeling group) mice separately. Pregnant mice in the modeling group were randomly grouped as follows: high/low-dose Shoutai Wan, high/low-dose Juyuan Jian, model (RPL), and positive control (dydrogesterone), with 10 mice in each group. Starting from the day after the detection of the vaginal plug, mice were administrated with drugs or an equal volume of normal saline by gavage for 10 consecutive days. After the intervention, the following indicators were measured. ① Macroscopic evaluation: general conditions, uterine wet weight, embryo loss rate, four coagulation parameters [prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), and thrombin time (TT)], and peripheral blood estradiol (E2) and progesterone (Pg) levels. The decidua with embryos was stained with hematoxylin-eosin (HE) and evaluated by transmission electron microscopy (TEM). The expression of B-cell lymphoma-2 (Bcl-2), vascular endothelial growth factor (VEGF), angiotensin Ⅱ (AngⅡ), matrix metalloproteinase-2 (MMP-2), interleukin-6 (IL-6), leukemia inhibitory factor (LIF), CXC chemokine ligand 12 (CXCL12), and microtubule-associated protein 1 light chain 3 homolog (LC3)Ⅰ/Ⅱ was quantified by Western blot. ② Mechanism analysis at the single-cell level: The decidua with embryos from the blank, model, high-dose Shoutai Wan, and high-dose Juyuan Jian groups (6 mice per group, with 3 single-cell samples per group, totaling 24 mice) were analyzed by the BD Rhapsody™ platform, and the whole-cell atlas was drawn by uniform manifold approximation and projection (UMAP) dimensionality reduction clustering combined with the single-cell mouse cell atlas (scMCA). The differentially expressed genes (DEGs) and cell interaction networks were analyzed via Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and CellChat, and the protein-protein interaction (PPI) map of subtype cells was constructed. The CytoTRACE pseudo-temporal analysis was performed to explore the developmental trajectories of core immune cells (natural killer cells, NK cells) from maternal and fetal sources. Results① Pathological and Western blot results indicated that compared with the blank group, the RPL group showed an increase in the embryo loss rate (P<0.01), down-regulated expression of Bcl-2, LIF, MMP-2, and Vegf in the decidua with embryos (P<0.05), up-regulated protein levels of CXCL-12, AngⅡ, and IL-6 (P<0.05), blocked angiogenesis, apoptosis-inflammation imbalance, and coagulation dysfunction. Both prescriptions dose-dependently reduced the abortion rate and restored the angiogenesis-inflammation balance, and Shoutai pill showed superior performance in restoring the E2 level to the Pg level (P<0.05). ② Single-cell transcriptome analysis indicated that compared with the blank group, the RPL group showed differences in multiple key cell populations such as decidual cells, trophoblast cells, endothelial cells, erythroblasts, NK cells, and macrophages at the maternal-fetal interface. Immunity and angiogenesis were the key links in RPL. Compared with the RPL group, high-dose Shoutai Wan reversed the changes of NK cells in the embryonic layer (upregulating the mRNA levels of 17 genes and downregulating the mRNA levels of 29 genes) and macrophages (upregulating the mRNA levels of 117 genes and downregulating the mRNA levels of 53 genes) through the regulation of gene expression. High-dose Shoutai pill regulated the immune cells to affect unfolded proteins, cell adhesion, and programmed cell death, thereby promoting decidualization and angiogenesis and modulating embryo-membrane development. High-dose Juyuan Jian regulated the key subgroups of NK cells (up-regulating the mRNA levels of 9 genes and down-regulating the mRNA levels of 17 genes) and macrophages (up-regulating the mRNA levels of 110 genes and down-regulating the mRNA levels of 81 genes), which affected decidual inflammation and apoptosis and intervened in glycolysis. ③ The pseudo-temporal analysis and communication network indicated that the communication frequency of the RPL group decreased. High-dose Shoutai Wan restored maternal-fetal tolerance through pathways such as NKG2D, CDH5, GDF, and FASLG. High-dose Juyuan Jian enhanced the IL-6/LIFR/JAK/signal transducer and activator of transcription 3 (STAT3) and desmosome/SEMA6/tumor necrosis factor-like weak inducer of apoptosis (TWEAK) signaling to improve endometrial receptivity. The RPL group showed an increased proportion of toxic dNK7, a decreased proportion of reparative dNK4, and blocked embryo fNK1. High-dose Shoutai Wan down-regulated dNK7 and up-regulated dNK4. High-dose Juyuan Jian inhibited the terminal differentiation of dNK7 and up-regulated LILRB1, thus restoring the balance of cytotoxicity and repair. ConclusionBoth the kidney-tonifying and spleen-invigorating methods are effective in treating RPL. NK and macrophages are the key immune cells in the interaction between the embryo and the membrane. The kidney-tonifying method (Shoutai Wan) has an advantage in regulating the phenotypes of unfolded protein, cell adhesion, and programmed cell death, and shows expression characteristics closer to the physiological state in the regulation of NKG2D and CDH5 signals. The spleen-invigorating method (Juyuan Jian) has an advantage in regulating epithelial-mesenchymal transition (EMT), angiogenesis, and glycolysis and shows higher communication intensity in the IL-6 and LIFR pathways.
2.Traditional Chinese Medicine Regulates Related Signaling Pathways to Prevent and Control Breast Cancer and Precancerous Lesions: A Review
Yifei ZENG ; Di ZHAO ; Junyue WANG ; Mengjie WANG ; Yubo GUO ; Yu ZHOU ; Dongxiao ZHANG ; Wenjie ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):290-301
Breast cancer has become the malignant tumor with the highest incidence rate among women, seriously threatening the life and health of women all over the world. The pathogenic factors and development mechanisms of breast cancer are complex and diverse. The development of breast cells from ordinary hyperplasia to atypical hyperplasia, and from pre-cancerous lesions to cancerous lesions, is a long-term progressive process. Therefore, early screening and prevention of breast cancer is particularly important. Western medicine has a relatively mature treatment program for breast cancer, which is mainly based on surgery and systemic treatment, whereas the ensuing complications and adverse reactions often bring a heavy burden to patients. For the precancerous lesions of breast cancer, surgery is also the mainstay of treatment. In recent years, traditional Chinese medicine (TCM) has increasingly highlighted its advantages in the prevention and treatment of breast cancer. Increasing studies have shown that in the prevention and treatment of breast cancer and pre-cancerous lesions, TCM compound prescriptions, single herbs or herb pairs, and active components are able to regulate a variety of intracellular signaling pathways through multi-targets to inhibit the proliferation and invasion, promote the apoptosis and autophagy of tumor cells, and regulate the cell cycle and the immune microenvironment, thus exerting anti-tumor effects. At the same time, they can significantly attenuate the toxic side effects of radiotherapy and drug resistance of patients. However, the specific mechanisms of TCM in the prevention and treatment of breast cancer and precancerous lesions have not been fully clarified. The available studies are tanglesome regarding the TCM inhibition of tumor development through the regulation of classical signaling pathways such as phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR), Wnt/β-catenin, and Notch, which still need to be verified by a large number of clinical and experimental studies. Therefore, this paper reviews the research progress in the prevention and treatment of breast cancer and precancerous lesions by TCM through interfering with the relevant signaling pathways in recent years, aiming to summarize the possible mechanisms of TCM in the prevention and treatment of breast cancer and provide references for subsequent studies.
3.Literature Analysis and Validity Assessment for Animal Models of Attention Deficit and Hyperactive Disorder
Wangyue LIAO ; Shuang LEI ; Xuan LI ; Min GUO ; Ruoran ZHOU
Laboratory Animal and Comparative Medicine 2026;46(1):66-80
Attention deficit and hyperactive disorder (ADHD) is the most common neurodevelopmental disorder of childhood. It seriously impairs academic achievement, social interaction, and vocational development, and increases the risk of accidental injury and substance abuse. In some cases, the symptoms may also exert an indirect disruptive effect on public order. Its aetiology involves interactions among genetic, perinatal environmental, and psychosocial factors that cannot be fully disentangled by single clinical studies. Therefore, a systematic evaluation of existing animal models is essential for revealing pathophysiology and developing novel therapies. Using the keywords "attention deficit and hyperactive disorder", "models, animal", "validity", and their English equivalents, we systematically searched PubMed, Web of Science, CNKI, and Wanfang for publications from 2000 to 2025 (retrieving 328 publications) and added further references by citation tracking. Eighty-six rodent ADHD models that provided detailed construction protocols, behavioural assessments, neurobiological mechanisms, or pharmacological data were included and classified into spontaneous genetic, genetically engineered, and environmentally induced paradigms. Their face, construct, and predictive validity were compared. Among spontaneous genetic models, spontaneously hypertensive rats reproduce hyperactivity, impulsivity, and stimulant responses well, yet hypertension and sex differences limit specificity. Acallosal mouse strains link corpus callosum absence to ADHD-like behaviours, but neurotransmitter studies remain scarce. Genetically engineered rodents—including dopamine transporter, neurokinin-1 receptor and mediator complex subunit 23 knockout or conditional gene knockout lines—precisely dissect dopaminergic, noradrenergic, synaptic, or epigenetic pathways, yet generally lack full phenotypic coverage, social-deficit modelling, and comorbidity representation, and are accompanied by adverse effects such as growth retardation or ocular defects. Environmentally induced models employ lead, polychlorinated biphenyls, alcohol, nicotine exposures, 6-hydroxydopamine lesions, neonatal hypoxia, early social isolation, or maternal stress to recapitulate core symptoms. However, dose-schedule standardisation is lacking. Behavioural reversibility diverges from clinical persistence, and non-specific phenotypes such as anxiety or depression are common. Overall, no single paradigm simultaneously achieves high validity across all three dimensions. Currently, ADHD models have progressed from single-factor simulations to multidimensional evaluation, yet significant gaps remain in genetic-background standardisation, sex differences, cross-species translation, and syndrome-differentiation modelling under traditional Chinese medicine. Future directions should integrate genetic, environmental, and epigenetic interactions, establish life-span validation systems, and incorporate computational neuroscience alongside integrative Chinese-Western strategies to enhance clinical relevance and translational utility, thereby providing robust evidence-based support for mechanistic elucidation, drug screening and precision intervention in ADHD.
4.Exploring on Quality Evaluation Methods of Clinical Case Reports in Traditional Chinese Medicine Based on China Clinical Cases Library of Traditional Chinese Medicine
Kaige ZHANG ; Feng ZHANG ; Bo ZHOU ; Haimin CHEN ; Yong ZHU ; Changcheng HOU ; Liangzhen YOU ; Weijun HUANG ; Jie YANG ; Guoshuang ZHU ; Shukun GONG ; Jianwen HE ; Yang YE ; Yuqiu AN ; Chunquan SUN ; Qingjie YUAN ; Buman LI ; Xingzhong FENG ; Kegang CAO ; Hongcai SHANG ; Jihua GUO ; Xiaoxiao ZHANG ; Zhining TIAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):271-276
As the core vehicle for preserving and transmitting traditional Chinese medicine(TCM) academic thought and clinical experience, the establishment of a robust quality evaluation system for TCM clinical case reports is a crucial component in the current standardization and modernization of TCM. Based on the practical experience of constructing the China Clinical Cases Library of Traditional Chinese Medicine by the China Association of Chinese Medicine, this study conducted a comprehensive analysis of critical challenges, including insufficient authenticity and unfocused evaluation criteria. It proposed a three-dimensional evaluation framework grounded in the structure-process-outcome logic, encompassing three dimensions of authenticity and standardization, characteristics and advantages, application and translational impact. This framework integrated 12 key evaluation indicators in a systematic manner. The model preserved the academic characteristics of TCM syndrome differentiation and treatment, while aligning with modern scientific research standards, achieving a balance between individualized TCM experience and standardized evaluation. Concurrently, this study provided theoretical foundations and methodological guidance for evaluating the quality of TCM clinical cases, contributing significantly to the inheritance of TCM knowledge, evidence-based practice, and the reform of talent evaluation mechanisms.
5.Association of 24 hour physical activity index with screening myopia and obesity in school aged children
ZHOU Keyi, LIU Su, MAIHELIYAKEZI Tuersunniyazi, GUO Manyu, YU Hongjie, SHI Huijing
Chinese Journal of School Health 2026;47(2):203-207
Objective:
To develop a 24 hour physical activity index for school aged children, and to analyze the association of 24 hour physical activity index with screening myopia and obesity, so as to provide a more effective assessment tool for student health risk screening.
Methods:
From April to June 2024, a total of 451 students in Grades 3-4 from two monitored primary schools in Shanghai were selected using stratified cluster random sampling method. Data on eight core indicators, including daily moderate to vigorous physical activity time, total physical activity time, outdoor activity time, screen time, sleep duration, sleep efficiency, social jetlag and daytime sleepiness, were collected through questionnaires and accelerometer monitoring. Each indicator was standardized and synthesized into a 0-80 point school aged children s 24 hour physical activity index. Spearman correlation analysis and t-test were used to assess consistency between questionnaire and accelerometer derived indices. Multivariate Logistic regression was applied to analyze the association of strength of the composite index with single behavior indicators in screening myopia and overweight/obesity.
Results:
The compliance rates were higher for moderate to vigorous physical activity time and screen time (50.8%, 98.7%), while the compliance rate for outdoor activity time was only 42.6%, and that for sufficient sleep duration was merely 10.2%. There was no significant difference between the total scores derived from the questionnaire and accelerometer methods (45.13±5.83, 45.05±6.87, t=0.29, P >0.05), but they showed a strong positive correlation ( r=0.58, P <0.01). Multivariate Logistic regression revealed that adjusting for individual behaviors such as grade, gender and both parents being myopic, among single behavior indicators of 24 hour physical activity index, only insufficient outdoor activity time was significantly associated with screening myopia among school aged children ( OR=1.50, 95%CI =1.01-2.21); the detection risk of screening myopia and obesity in the low index group were higher than those in the high index group ( OR=2.47, 95%CI =1.02-5.96; OR=16.63,95%CI = 5.99- 46.20) (all P <0.05).
Conclusion
The developed 24 hour physical activity index for school aged children demonstrates good measurement accuracy and shows stronger associations with screening myopia and obesity than single behavior indicator.
6.Pathogenesis of focal segmental glomerulosclerosis:from podocyte injury to precision therapy
Academic Journal of Naval Medical University 2025;46(10):1344-1349
Focal segmental glomerulosclerosis(FSGS)is a critical glomerular disease characterized by a complex pathogenesis involving podocyte injury,genetic variations,and immune dysregulation.Podocyte injury plays a central role in FSGS development,encompassing abnormalities in cytoskeletal structure and dynamics,dysfunction of cell adhesion,activation of programmed cell death pathways,and oxidative stress.In recent years,advancements in molecular biology and genomic technologies have deepened the understanding of FSGS pathogenesis,with notable progress in podocyte biology,genetics,and signaling pathway regulation.Genetic factors contribute significantly to FSGS,with studies on monogenic mutations,epigenetic regulatory mechanisms,and non-genetic mechanisms such as the circulating factor hypothesis and immune dysregulation enriching the molecular pathology of FSGS.Based on these mechanistic insights,researchers are exploring precision therapeutic targets,including podocyte-protective strategies,anti-fibrotic and anti-inflammatory approaches,and gene-and biomarker-guided therapies,to improve treatment outcomes.This review summarizes recent advances in the pathogenesis of FSGS,aiming to provide a theoretical framework and new insights for precision diagnosis and targeted therapy of this disease.
7.Removal of plaque biofilm by a 3D-printed precision pericoronal irrigator for impacted wisdom teeth:an in vitro study
Xiaoting WANG ; Zhenxin HUANG ; Weiwei GUO ; Jianye SONG ; Liang ZHOU ; Rouyi WEI ; Kun WANG
Journal of Army Medical University 2025;47(7):734-741
Objective To prepare an advanced irrigation device equipped with a specialized nozzle designed to access the pericoronal blind sac of impacted wisdom teeth,and evaluate its efficacy in plaque removal.Methods 3D-printer was utilized to develop pericoronal models simulating mandibular wisdom teeth with high vertical impaction and varying soft tissue coverage(0%,25%,50%,and 75%,respectively)and personalized pericoronal irrigator.The obtained pericoronal models were randomly divided into 3 groups,treated with personalized precision pericoronal flushing,routine flushing and brushing,respectively,and,under a pressures of 45 psi or 75 psi(1 psi=6 894.76 Pa),were performed to clean the test surfaces of plaque-biofilm-obstructed wisdom teeth,respectively.Images of the tooth surfaces were analyzed before and after cleaning,and the cleaning rate was calculated using Python software.Results For wisdom teeth with 50%and 75%pericoronal soft tissue coverage,at a pressure of 45 psi,the precision pericoronal flushing group demonstrated significantly higher cleaning rate when compared to the brushing and routine flushing groups(P<0.05).At 0%and 25%pericoronal soft tissue coverage,the precision pericoronal flushing and brushing groups exhibited significantly higher cleaning rates than the routine flushing group(P<0.05).Furthermore,at 75 psi,the precision pericoronal flushing group achieved a significantly higher cleaning rate across all 4 levels of pericoronal soft tissue coverage than both the routine flushing and brushing groups(P<0.05).Notably,the precision pericoronal flushing group at 75 psi obtained significantly higher cleaning rate than at 45 psi across 0%,25%,and 50%pericoronal soft tissue coverage(P<0.05).Conclusion The precision pericoronal irrigation device demonstrates significantly superior cleaning efficacy for vertically impacted mandibular third molars compared to conventional irrigation devices and manual tooth brushing.It is an effective device for preventing and treating pericoronitis.
8.Prediction of risk for acute kidney injury and its progression to mortality in obese patients admitted to ICU postoperatively
Qiang LI ; Guo MU ; Wenzhang WANG ; Jie YIN ; Xuan YU ; Bin LU ; Qian LI ; Jun ZHOU
Journal of Army Medical University 2025;47(10):1110-1125
Objective To develop a machine learning-based risk prediction model for postoperative acute kidney injury(AKI)and a model for mortality in obese patients admitted to intensive care unit(ICU)in order to improve early warning and prognostic evaluation to support clinical decision-making.Methods Data of obese postoperative ICU patients were retrospectively retrieved from the MIMIC-Ⅳ and eICU databases for statistical analysis.Ultimately,2 520 patients(670 from MIMIC-Ⅳ and 1 850 from eICU databases)were included to build the risk prediction models for AKI and mortality.The data included demographic information,vital signs,laboratory findings,surgical types,comorbidities,and medication use.After data cleaning and preprocessing,Boruta feature selection was applied,followed by the construction of prediction models using 7 machine learning algorithms,that is,Gradient Boosting Machine(GBM),Generalized Linear Model(GLM),k-Nearest Neighbors(KNN),Na?ve Bayes(NB),Neural Network(NNET),Support Vector Machine(SVM),and XGBoost.Model performance was evaluated through cross-validation and external validation.Results In the risk prediction models of AKI,the SVM model achieved the highest AUC value of 0.80 in the testing set and 0.71 in the external validation test.For the risk prediction models of mortality,the GBM model outperformed others in the prediction,attaining an AUC value of 0.91 in the testing set.Conclusion Risk predictive models for postoperative AKI and mortality in obese ICU patients are successfully constructed,and are valuable tools for clinicians to optimize early intervention and improve clinical outcomes for the patients.
9.Change in serum uric acid level and its influencing factors among military personnel during long-term maritime mission
Lifeng SHI ; Yan WU ; Guangyong WANG ; Shaoyu ZHANG ; Fang WANG ; Tao GUO ; Guangman TANG ; Lan LI ; Yibing ZHOU
Journal of Army Medical University 2025;47(12):1284-1290
Objective To investigate the characteristics of changes in blood uric acid(UA)and detection rate of hyperuricemia(HUA)among officers and soldiers during long-term maritime missions,as well as their related influencing factors.Methods A total of 100 servicemen were randomly selected from 240 officers and soldiers who will participate in a long-distance voyage mission.Their general information,including age,education level,administrative position,years of service on board,and department,was surveyed.Their annual data of physical examination were retrospectively analyzed and compared with the results of another 335 shore-based servicemen during the same period.On mission day 10(D10)and day 50(D50),the venous blood samples were collected from the participants to synchronously measure blood UA level and body composition indicators(body fat mass,BMI,fat percentage,fat mass,muscle mass,and muscle percentage).Additionally,on D50,Self-Rating Scale of Sleep(SRSS)and Symptom Checklist-90(SCL-90)were employed to survey their conditions.Seventy service members were randomly selected from the 100 participants to engage in aerobic exercise.The changes in UA level and detection rate of HUA among the mission personnel were analyzed,along with their influencing factors.Results The UA level and HUA detection rate in long-term navigation personnel during concurrent annual physical examinations were significantly lower than those in shore-based personnel(P<0.01).Compared to pre-voyage physical examination results,the UA level and HUA detection rate in long-term navigation personnel were significantly increased from mission day D10(P<0.001).Compared to the values at D10,the UA level and HUA detection rate at D50 showed significant decreases(P<0.05),and then essentially returned to pre-mission examination levels(P>0.05).Aged<32 years was an independent risk factor for new-onset HUA at mission D10(P<0.05).<32 years old and aerobic exercise during the voyage were independent influencing factors for HUA outcome(P<0.05).Conclusion Serum UA level and HUA detection rate among officers and soldiers participating in long-term maritime missions are relatively low before departure,but in significant increases during the early stages of the mission,particularly among those aged<32 years.Scientific aerobic exercise during the mission period helps reduce UA level and HUA detection rate,playing a crucial role in guaranteeing physical and mental health.
10.Role and mechanism of a novel fusion gene RELCH-RET in driving malignant transformation of human bronchial epithelial cells:a preliminary study
Xiaogang ZHOU ; Xianglin HAO ; Jiying XIA ; Zhimin HUANGFU ; Wanlei FU ; Yangfan LYU ; Qiaonan GUO
Journal of Army Medical University 2025;47(13):1444-1453
Objective To investigate the role and primary mechanism of a novel fusion gene RELCH-RET in driving the malignant transformation of normal human bronchial epithelial(HBE)cells.Methods Based on retrospective clinical data from 456 non-small cell lung cancer(NSCLC)patients admitted in the Second Affiliated Hospital of Army Medical University from January 2019 to June 2022,a fusion gene,RELCH-RET,was identified as a research target.Three cell models were established:negative control(HBE VC,transfected with empty lentiviral vector),RET control(HBE RET,transfected with lentiviral overexpression vector of Flag-RET),and experimental group(HBE RELCH-RET,transfected with lentiviral overexpression vector of Flag-RELCH-RET).MTS assay and Transwell assay were used to detect cell proliferation and migratory and invasive abilities.In vivo tumorigenicity of the 3 cell models was assessed in 15 female non-obese diabetic/severe combined immunodeficiency(NOD/SCID)mice(SPF grade,4 weeks old,weighing 15.1±0.4 g)via subcutaneous xenograft experiments,with 5 animals in each group.Western blotting was employed to detect the autophosphorylation of RET(Y905)and the phosphorylation of downstream signaling proteins ERK1/2,EGFR(Y845)and STAT3(Y705).Dimerization and multimerization status of RELCH-RET were analyzed by chemical cross-linking(DTME treatment)in combination with Western blotting,with the reversibility being confirmed through de-cross-linking experiments.Results There were 3 cases carrying RELCH-RET fusion gene screened out from the 469 NSCLC patients.Compared with the HBE VC and HBE RET groups,the HBE RELCH-RET group exhibited significantly enhanced cell proliferation(P<0.01),and acquired migratory and invasive abilities(P<0.01),while the control groups did not demonstrate the abilities.In the mouse xenograft tumor model,HBE cells stably expressing RELCH-RET developed significant tumor nodules(P<0.001),whereas the control groups(empty vector and wild-type RET)failed to exhibit detectable tumor growth.Western blotting revealed that RELCH-RET could induce the autophosphorylation of the RET tyrosine residue(Y905)and significantly up-regulate the phosphorylation levels of ERK1/2,EGFR(Y845),and STAT3(Y705)proteins.Chemical cross-linking combined with Western blot analysis demonstrated that RELCH-RET formed a dimer(~170 kDa)in HBE cells,which is reversibly dissociated into monomers upon decross-linking treatment.Conclusion The novel fusion gene RELCH-RET,promotes ligand-independent dimerization/oligomerization,thereby mediating RET autophosphorylation,subsequently activates the downstream typical RET signaling pathway and ultimately drives the malignant transformation of normal HBE cells.


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