1.Application of radiomics combined with machine learning algorithms for preoperative prediction of perineural invasion in oral squamous cell carcinoma
MENG Xiangze ; YUAN Ying ; YANG Xi
Journal of Prevention and Treatment for Stomatological Diseases 2026;34(5):456-470
Objective:
To explore the value of contrast-enhanced computed tomography (CT) radiomics combined with machine learning algorithms in the preoperative prediction of perineural invasion (PNI) in oral squamous cell carcinoma (OSCC), aiming to provide evidence for assisting clinical treatment decision-making.
Methods:
This study was approved by the Ethics Committee of the Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine. A total of 250 OSCC patients confirmed by postoperative pathology were included, comprising 128 PNI-positive and 122 PNI-negative cases. The dataset was randomly divided into training (n=175), validation (n=38), and independent testing (n=37) sets in a ratio of 7:1.5:1.5. Regions of interest were delineated on preoperative images, and radiomic features were extracted. After dimensionality reduction and feature selection using methods like Least Absolute Shrinkage and Selection Operator (LASSO) regression, multiple machine learning models, including support vector machine (SVM), random forest, Light gradient boosting machine (LightGBM), and a Stacking ensemble model, were constructed. Model performance was evaluated using metrics such as the area under the receiver operating characteristic curve (AUC), sensitivity, calibration curves, and decision curve analysis (DCA). Model interpretability was analyzed using Shapley additive explanations (SHAP) and grouped permutation feature importance analysis.
Results :
Among the 250 samples analyzed, the LightGBM model based on radiomics demonstrated the best performance on the independent test set, with an AUC of 0.781, outperforming models like SVM (AUC = 0.730) and Random Forest (AUC = 0.691), as well as clinical models (AUCs ranging 0.549-0.711). The LightGBM model showed good calibration (Brier score 0.198), and DCA indicated high clinical net benefit over a wide threshold probability range. Paired DeLong tests revealed no statistically significant differences in AUC between the ensemble (Stacking) model and the corresponding best-performing radiomics-based model. SHAP analysis and grouped permutation feature importance analysis further indicated that the primary discriminative information for the model came from radiomic texture features.
Conclusion
The LightGBM model based on contrast-enhanced CT radiomics demonstrated good discriminative ability for preoperative prediction of PNI in OSCC. In the independent test set, it achieved the highest AUC. This model holds promise as a non-invasive auxiliary tool for preoperative risk assessment. Given the limited sample size of the independent test set, these results require further validation in larger cohorts and external datasets.
2.Clinical efficacy of curettage in treating cesarean scar pregnancy:a randomized controlled trial
Ying YANG ; Lu ZHOU ; Li LUO ; Xi XIONG ; Zhengqiong CHEN
Journal of Army Medical University 2025;47(9):989-994
Objective To compare the safety and efficacy of curettage and their combination with uterine artery embolization(UAE)in the treatment of cesarean scar pregnancy(CSP)patients with a low score(≤4)in the ultrasound quantification scoring system.Methods Based on our inclusion and exclusion criteria of this randomized controlled study,the women with CSP who had an ultrasonic quantitative score≤4 and were treated in our department from May 2020 to August 2023 were enrolled,and then randomly divided into a curettage group(n=48)and a UAE combination group(n=47)in a ratio of 1∶1.General information,intraoperative conditions,and use of rescue measures within 3 months after operation were collected in the 2 groups of patients.All the patients were followed up until October 2024 to observe the pregnancy outcomes and determine the impact on the menstrual volume after the resumption of normal menstruation.Results The patients from the both groups completed the follow-up.Except for the maximum gestational sac diameter,there were no significant differences in other baseline data between the 2 groups,and the curettage group had notably more patients having a gestational sac diameter≤25 mm than the combination group[37(77.1%)vs 27(57.4%),P<0.05].No statistical differences were observed between the 2 groups in the intraoperative bleeding volume and use of rescue measures within 3 months after surgery.The combination group had obviously more patients with reduced menstrual volume after the resumption of normal menstruation than the dilation and curettage group[30(63.8%)vs 13(27.1%),P<0.001].There were no statistically differences in pregnancy outcomes and the number of days to resume menstruation between the 2 groups.Conclusion For CSP patients with a score of≤4 in the ultrasound quantification scoring system,curettage show no significant difference in therapeutic effectiveness,and even have better efficacy and safety when compared with curettage combined with UAE.
3.Hippocampal Extracellular Matrix Protein Laminin β1 Regulates Neuropathic Pain and Pain-Related Cognitive Impairment.
Ying-Chun LI ; Pei-Yang LIU ; Hai-Tao LI ; Shuai WANG ; Yun-Xin SHI ; Zhen-Zhen LI ; Wen-Guang CHU ; Xia LI ; Wan-Neng LIU ; Xing-Xing ZHENG ; Fei WANG ; Wen-Juan HAN ; Jie ZHANG ; Sheng-Xi WU ; Rou-Gang XIE ; Ceng LUO
Neuroscience Bulletin 2025;41(12):2127-2147
Patients suffering from nerve injury often experience exacerbated pain responses and complain of memory deficits. The dorsal hippocampus (dHPC), a well-defined region responsible for learning and memory, displays maladaptive plasticity upon injury, which is assumed to underlie pain hypersensitivity and cognitive deficits. However, much attention has thus far been paid to intracellular mechanisms of plasticity rather than extracellular alterations that might trigger and facilitate intracellular changes. Emerging evidence has shown that nerve injury alters the microarchitecture of the extracellular matrix (ECM) and decreases ECM rigidity in the dHPC. Despite this, it remains elusive which element of the ECM in the dHPC is affected and how it contributes to neuropathic pain and comorbid cognitive deficits. Laminin, a key element of the ECM, consists of α-, β-, and γ-chains and has been implicated in several pathophysiological processes. Here, we showed that peripheral nerve injury downregulates laminin β1 (LAMB1) in the dHPC. Silencing of hippocampal LAMB1 exacerbates pain sensitivity and induces cognitive dysfunction. Further mechanistic analysis revealed that loss of hippocampal LAMB1 causes dysregulated Src/NR2A signaling cascades via interaction with integrin β1, leading to decreased Ca2+ levels in pyramidal neurons, which in turn orchestrates structural and functional plasticity and eventually results in exaggerated pain responses and cognitive deficits. In this study, we shed new light on the functional capability of hippocampal ECM LAMB1 in the modulation of neuropathic pain and comorbid cognitive deficits, and reveal a mechanism that conveys extracellular alterations to intracellular plasticity. Moreover, we identified hippocampal LAMB1/integrin β1 signaling as a potential therapeutic target for the treatment of neuropathic pain and related memory loss.
Animals
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Laminin/genetics*
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Hippocampus/metabolism*
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Neuralgia/metabolism*
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Cognitive Dysfunction/etiology*
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Male
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Peripheral Nerve Injuries/metabolism*
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Extracellular Matrix/metabolism*
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Integrin beta1/metabolism*
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Pyramidal Cells/metabolism*
;
Signal Transduction
4.Application of microflow imaging (MFI) technology based on Image Pro Plus software in BI-RADS 4 breast nodules and correlation between MFI parameters and Ki-67 expression
Fan YANG ; Chunwei LIU ; Dai ZHANG ; Ying WANG ; Hailing WANG ; Xi WEI
Chinese Journal of General Surgery 2025;40(7):551-557
Objective:To evaluate the diagnostic value of micro flow imaging (MFI) supported by Image Pro Plus (IPP) software for BI-RADS classification of breast nodules and the correlation between its parameters and Ki-67 expression level.Methods:A total of 205 cases of breast solid nodules with ultrasound BI-RADS classification of 4 categories were enrolled in this study. BI-RADS classification, CDFI score, MFI score, CDFI-IPP score and MFI-IPP score by IPP software were performed for each nodule. Ki-67 immunohistochemical staining of invasive ductal carcinoma was performed to analyze the correlation between MFI-IPP-VI and Ki-67 expression levels.Results:The combined BI-RADS classification for diagnosis of malignant nodules had the largest area under the ROC curve, which was significantly higher than that of BI-RADS, MFI-IPP-VI, MFI score, CDFI-IPP-VI, and CDFI score (all P<0.05). The combined BI-RADS classification had the best diagnostic efficacy for the diagnosis of malignant nodules with a sensitivity of 96.4%, specificity of 65.6% and accuracy of 82.4%. In addition, the agreement between experienced and inexperienced sonographers applying MFI-IPP-VI was higher than MFI (Kappa=0.780 vs. Kappa=0.743). The mean value of Ki-67 in invasive ductal carcinoma was 21.93%±16.37%, and Spearman's correlation analysis showed a positive correlation between MFI-IPP-VI and Ki-67 expression level ( r=0.815, P<0.05). Conclusions:MFI-IPP can assist sonographers to classify breast nodules by BI-RADS, providing an effective basis for puncture biopsy. MFI-IPP-VI offers the possibility of noninvasively predicting the expression level of Ki-67 in invasive ductal carcinoma.
5.Preliminary investigation into the role of Sneathia Sanguinegens in unexplained recurrent spontaneous abortion
Fu-ju ZHAO ; Xian-yang HU ; Lu LIU ; Xi-xi HUANG ; Fei-fei WANG ; Jing GAO ; Mei-rong DU ; Chun-mei YING
Fudan University Journal of Medical Sciences 2025;52(1):99-106,138
Objective To investigate the role of Sneathia sanguinegens(S.sanguinegens)in the development of unexplained recurrent spontaneous abortion(URSA).Methods A case-control study was conducted to analyze the vaginal flora characteristics of 65 patients with URSA and 18 healthy controls through 16S rRNA gene sequencing.Toxicity profile of S.sanguinegens on human cervical cancer cells(ME-180),human umbilical vein endothelial cells(HUVEC)and human placental choriocarcinoma cells(JEG-3)was analyzed at the cellular level to assess the mechanism of it in adverse pregnancy outcomes.And S.sanguinegens was used to infect C57BL/6J mice to explore the toxic effect on living organisms.Results The relative abundance of Sneathia was increased in patients with URSA compared with healthy controls.It was positively correlated with the number of miscarriages,and was attributed to S.sanguinegens.We also found that S.sanguinegens damaged ME-180,JEG-3 and HUVEC cells.The degree of cellular damage was related to the level of S.sanguinegens added.Intravenous infection with S.sanguinegens caused inflammatory damage in several organs and extramedullary hematopoiesis in the spleen.Conclusion S.sanguinegens is closely related to URSA and should be emphasized in patients with high vaginal bacterial load.
6.Establishment of a rat model of comorbid chronic inflammatory pain and depression using different doses of complete Freund's adjuvant
Tian WANG ; Pu YANG ; Xi ZHANG ; Xin LI ; Wenjing HUANG ; Guangmei ZHENG ; Xinyu HUANG ; Zhiyi XU ; Ying HUANG ; Lili YIN ; Shengyong SU
Acta Laboratorium Animalis Scientia Sinica 2025;33(5):633-643
Objective To compare the success rate and stability of rat models of comorbid chronic pain and depression induced by different doses of complete Freund's adjuvant(CFA).Methods Sixty SD rats were divided randomly into a control group,low-dose CFA group(CFA-L),and high-dose CFA group(CFA-H)(n=20 rats per group).Rats in the CFA-L and CFA-H groups were injected with 50 and 100 μL CFA,respectively,and rats in the control group were injected with 0.9%sodium chloride solution.The general state,body weight,mechanical withdrawal threshold(MWT),and thermal withdrawal latency(TWL)were observed at 0,7,14,21,and 28 days after modeling.Depressive behavior was evaluated using the open field test(OFT),forced swim test(FST),and tail suspension test(TST).Glutamate(Glu)and γ-aminobutyric acid(GABA)levels in the anterior cingulate cortex were detected by enzyme-linked immunosorbent assay.Brain-derived neurotrophic factor(BDNF)expression in the anterior cingulate cortex was detected by immunohistochemistry,and pathological changes in the anterior cingulate cortex were observed by HE staining.Results(1)Regarding the general condition of the rats,the left ankle joint and toes were obviously red and swollen in the CFA-L and CFA-H groups on the 7th day after modeling,and the swelling was more severe in the CFA-H group.The redness and swelling of the left hind foot and ankle joint and toes gradually recovered in the CFA-L group on days 14,21,and 28 after modeling,but were still obvious in the CFA-H group,and the water and food intake decreased.(2)The body mass was significantly lower in rats in the CFA-H group compared with those in the blank and CFA-L groups on days 14,21,and 28 after modeling(P<0.05,P<0.05).(3)Regarding pain-related behavior,the MWT and TWL were significantly decreased in the CFA-L and CFA-H groups on the 7th and 14th days after modeling,compared with the control group(P<0.05,P<0.05).On day 21 after modeling,MWT was significantly lower in the CFA-H group than in the blank and CFA-L groups(P<0.05,P<0.05),and TWL was significantly lower in the CFA-L and CFA-H groups than in the blank group(P<0.05,P<0.05).On day 28 after modeling,MWT and TWL were significantly lower in the CFA-H group than in the blank and CFA-L groups(P<0.05,P<0.05).(4)In terms of depression-related behaviors,the total OFT movement distance was significantly lower in the CFA-H group than in the blank and CFA-L groups on day 7 after modeling(P<0.05,P<0.05).The total OFT distance and central dwell time were significantly lower in the CFA-H group than in the blank and CFA-L groups on days 14,21,and 28 after modeling(P<0.05,P<0.05),and the result in the FST and TST were significantly higher than in the blank and CFA-L groups(P<0.05,P<0.05).(5)Glu,GABA,and BDNF expression levels were significantly higher in the CFA-H group than in the blank and CFA-L groups(P<0.05,P<0.05),while GABA,Glu/GABA,and BDNF levels were significantly lower in the CFA-H group than in the blank and CFA-L groups(P<0.05,P<0.05,P<0.05).(6)The CFA-L group showed less damage in the anterior cingulate cortex,more pyramidal cells,more arranged cells,clear nucleoli,and a small number of cells with karyokynesis and deep staining.Compared with the CFA-L group,rats in the CFA-H group showed a disordered cell arrangement in the injured area of the anterior cingulate cortex,a large number of pyknotic and hyperchromatic neurons,significantly fewer or absent pyramidal cells,and vacuoles,red blood cells,and neurofibrillary tangles in the interstitial space.Conclusions Injection of CFA 100 μL can be used to establish a rat model of chronic inflammatory pain and depression,showing hyperalgesia,depression-like behavioral changes,changes in levels of Glu,GABA,and BDNF in the anterior cingulate cortex,and pathological changes in the anterior cingulate cortex,consistent with the pathophysiological characteristics of chronic pain and depression.
7.A Randomized Controlled Trial of Stone Needle Thermocompression and Massage for Treating Chronic Musculoskeletal Pain in the Shoulder and Back:A Secondary Analysis of Muscle Elasticity as a Mediator
Jingjing QIAN ; Yuanjing LI ; Li LI ; Yawei XI ; Ying WANG ; Cuihua GUO ; Jiayan ZHOU ; Yaxuan SUN ; Shu LIU ; Guangjing YANG ; Na YUAN ; Xiaofang YANG
Journal of Traditional Chinese Medicine 2025;66(9):935-940
ObjectiveTo evaluate the effectiveness of stone needle thermocompression and massage compared to flurbiprofen gel patch in relieving chronic musculoskeletal pain in the shoulder and back, and to explore the potential mediating mechanism through muscle elasticity. MethodsA total of 120 patients with chronic musculoskeletal pain in the shoulder and back were randomly assigned to either stone needle group or flurbiprofen group, with 60 patients in each. The stone needle group received stone needle thermocompression and massage for 30 minutes, three times per week; the flurbiprofen group received flurbiprofen gel patch twice daily. Both groups were treated for 2 weeks. Pain improvement, as the primary outcome, was assessed using the Global Pain Scale (GPS) at baseline, after 2 weeks of treatment, and again 2 weeks post-treatment. To explore potential mechanisms, a mediator analysis was conducted by measuring changes in superficial and deep muscle elasticity using musculoskeletal ultrasound at baseline and after the 2-week treatment period. ResultsThe stone needle group showed significantly greater pain relief than the flurbiprofen group 2 weeks post-treatment. After adjusting for confounders related to pain duration, the between-group mean difference was -8.8 [95% CI (-18.2, -0.7), P<0.05]. Part of the therapeutic effect was mediated by changes in deep muscle elasticity, with a mediation effect size of -1.5 [95% CI (-2.0, -0.9), P = 0.024], accounting for 17.9% of the total effect. ConclusionStone needle thermocompression and massage can effectively relieve chronic musculoskeletal pain in the shoulder and back, partly through a mediating effect of improved deep muscle elasticity.
8.PANoptosis: a New Target for Cardiovascular Diseases
Xin-Nong CHEN ; Ying-Xi YANG ; Xiao-Chen GUO ; Jun-Ping ZHANG ; Na-Wen LIU
Progress in Biochemistry and Biophysics 2025;52(5):1113-1125
The innate immune system detects cellular stressors and microbial infections, activating programmed cell death (PCD) pathways to eliminate intracellular pathogens and maintain homeostasis. Among these pathways, pyroptosis, apoptosis, and necroptosis represent the most characteristic forms of PCD. Although initially regarded as mechanistically distinct, emerging research has revealed significant crosstalk among their signaling cascades. Consequently, the concept of PANoptosis has been proposed—an inflammatory cell death pathway driven by caspases and receptor-interacting protein kinases (RIPKs), and regulated by the PANoptosome, which integrates key features of pyroptosis, apoptosis, and necroptosis. The core mechanism of PANoptosis involves the assembly and activation of the PANoptosome, a macromolecular complex composed of three structural components: sensor proteins, adaptor proteins, and effector proteins. Sensors detect upstream stimuli and transmit signals downstream, recruiting critical molecules via adaptors to form a molecular scaffold. This scaffold activates effectors, triggering intracellular signaling cascades that culminate in PANoptosis. The PANoptosome is regulated by upstream molecules such as interferon regulatory factor 1 (IRF1), transforming growth factor beta-activated kinase 1 (TAK1), and adenosine deaminase acting on RNA 1 (ADAR1), which function as molecular switches to control PANoptosis. Targeting these switches represents a promising therapeutic strategy. Furthermore, PANoptosis is influenced by organelle functions, including those of the mitochondria, endoplasmic reticulum, and lysosomes, highlighting organelle-targeted interventions as effective regulatory approaches. Cardiovascular diseases (CVDs), the leading global cause of morbidity and mortality, are profoundly impacted by PCD. Extensive crosstalk among multiple cell death pathways in CVDs suggests a complex regulatory network. As a novel cell death modality bridging pyroptosis, apoptosis, and necroptosis, PANoptosis offers fresh insights into the complexity of cell death and provides innovative strategies for CVD treatment. This review summarizes current evidence linking PANoptosis to various CVDs, including myocardial ischemia/reperfusion injury, myocardial infarction, heart failure, arrhythmogenic cardiomyopathy, sepsis-induced cardiomyopathy, cardiotoxic injury, atherosclerosis, abdominal aortic aneurysm, thoracic aortic aneurysm and dissection, and vascular toxic injury, thereby providing critical clinical insights into CVD pathophysiology. However, the current understanding of PANoptosis in CVDs remains incomplete. First, while PANoptosis in cardiomyocytes and vascular smooth muscle cells has been implicated in CVD pathogenesis, its role in other cell types—such as vascular endothelial cells and immune cells (e.g., macrophages)—warrants further investigation. Second, although pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs) are known to activate the PANoptosome in infectious diseases, the stimuli driving PANoptosis in CVDs remain poorly defined. Additionally, methodological challenges persist in identifying PANoptosome assembly in CVDs and in establishing reliable PANoptosis models. Beyond the diseases discussed, PANoptosis may also play a role in viral myocarditis and diabetic cardiomyopathy, necessitating further exploration. In conclusion, elucidating the role of PANoptosis in CVDs opens new avenues for drug development. Targeting this pathway could yield transformative therapies, addressing unmet clinical needs in cardiovascular medicine.
9.Mechanism of Kidney-tonifying and Liver-regulating Cyclical Therapy and Formula in Improving Endometrial Receptivity during "Implantation Window" in Rats with Polycystic Ovary Syndrome via miR-140-5p/VEGF Pathway
Quan LIU ; Yiqing HE ; Ying ZHANG ; Xi WANG ; Shuo YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):100-109
ObjectiveTo investigate the mechanism of kidney-tonifying and liver-regulating cyclical therapy and its formula in regulating endometrial receptivity during the "implantation window" in rats with polycystic ovary syndrome (PCOS). MethodsSix rats were randomly selected from 36 SPF SD female rats as the normal group, and the remaining rats were administered letrozole to induce a PCOS model. By using a random number method, the rats were divided into the following groups: normal group, model group, Xiaoyaosan group (11.97 g·kg-1), Sanzi Yangmo decoction group (28.35 g·kg-1), cyclical therapy group (11.97/28.35 g·kg-1), and aspirin group (8 × 10-3 mg·kg-1). After 12 days of continuous administration by gavage (equivalent to three estrous cycles), female and male rats were co-housed. On the fifth day of pregnancy, the number of blastocyst implantation in each group was counted. Hematoxylin-eosin (HE) staining was used to observe the pathological morphology of rat endometrial tissue. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of estradiol (E2) and progesterone (P) in rat serum. Western blot was used to detect the protein expression of vascular endothelial growth factor (VEGF), progesterone receptor (PR), estrogen receptor (ER), androgen receptor (AR), and integrin(ITG) αvβ3 in rat endometrial blood vessels. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of miR-140-5 P, VEGF, vascular endothelial growth factor receptor 2 (VEGFR2), PR, ER, AR, and ITGαvβ3 in rat endometrium. ResultsCompared with normal group, the estrous cycle of the rats in model group continued to be in the estrus interval and the estrous cycle lost regular changes. The endometrium was significantly thinner, the number of uterine glands and blood vessels were significantly reduced (P<0.01), and the pregnancy rate was significantly reduced. Compared with the model group, each drug group restored the regular estrous cycle to varying degrees, and the endometrial thickness and the number of blood vessels were significantly improved (P<0.01). The pregnancy rate of each drug group increased, and the effect of the cycle therapy group could reach the normal level. The results of molecular biology experiments showed that compared with the normal group, the levels of serum E2 and P in the model group were significantly decreased (P<0.01), the expression of VEGF, ER, PR and ITGαvβ3 protein was significantly decreased (P<0.05,P<0.01), the expression of AR protein was significantly increased (P<0.01), the expression of miR-140-5P and AR mRNA was significantly increased (P<0.01), and the expression of VEGF, VEGFR2, ER, PR and ITGαvβ3 mRNA was significantly decreased (P<0.01). Compared with model group, the serum E2 level in the Xiaoyaosan group was significantly increased (P<0.01).The levels of E2 and P in serum of rats in Sanzi Yangmo decoction group, cycle therapy group and aspirin group were significantly increased (P<0.01). The expression of AR protein in each drug group was significantly decreased (P<0.01). The expression of VEGF and ITGαvβ3 protein in Xiaoyaosan group was significantly increased (P<0.01). The expression of VEGF, ER and PR protein in Sanzi Yangmo decoction group was increased to varying degrees (P<0.05,P<0.01). The expression of VEGF, PR, ER and ITGαvβ3 protein in the cycle therapy group and the aspirin group increased to varying degrees (P<0.05,P<0.01). The expression of miR-140-5P and AR mRNA in each drug group was significantly decreased (P<0.01). The expression of VEGF, VEGFR2, ER, PR and ITGαvβ3 mRNA in each drug group increased to varying degrees (P<0.05,P<0.01). Compared with Xiaoyaosan group and Sanzi Yangmo decoction group, the expression of miR-140-5P, VEGFR2, ER, PR, AR and ITGαvβ3 mRNA in the cycle therapy group were significantly different (P<0.05,P<0.01). ConclusionThe kidney-tonifying and liver-regulating cyclical therapy may reduce the activity of miR-140-5P, target the upregulation of VEGF expression, mediate angiogenesis, and promote endometrial angiogenesis, thereby playing a synergistic role in improving endometrial receptivity in PCOS-induced infertility. Its efficacy in increasing pregnancy rates surpasses that of Xiaoyaosan or Sanzi Yangmo decoction used alone.
10.Mechanism of Kidney-tonifying and Liver-regulating Cyclical Therapy and Formula in Improving Endometrial Receptivity during "Implantation Window" in Rats with Polycystic Ovary Syndrome via miR-140-5p/VEGF Pathway
Quan LIU ; Yiqing HE ; Ying ZHANG ; Xi WANG ; Shuo YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):100-109
ObjectiveTo investigate the mechanism of kidney-tonifying and liver-regulating cyclical therapy and its formula in regulating endometrial receptivity during the "implantation window" in rats with polycystic ovary syndrome (PCOS). MethodsSix rats were randomly selected from 36 SPF SD female rats as the normal group, and the remaining rats were administered letrozole to induce a PCOS model. By using a random number method, the rats were divided into the following groups: normal group, model group, Xiaoyaosan group (11.97 g·kg-1), Sanzi Yangmo decoction group (28.35 g·kg-1), cyclical therapy group (11.97/28.35 g·kg-1), and aspirin group (8 × 10-3 mg·kg-1). After 12 days of continuous administration by gavage (equivalent to three estrous cycles), female and male rats were co-housed. On the fifth day of pregnancy, the number of blastocyst implantation in each group was counted. Hematoxylin-eosin (HE) staining was used to observe the pathological morphology of rat endometrial tissue. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of estradiol (E2) and progesterone (P) in rat serum. Western blot was used to detect the protein expression of vascular endothelial growth factor (VEGF), progesterone receptor (PR), estrogen receptor (ER), androgen receptor (AR), and integrin(ITG) αvβ3 in rat endometrial blood vessels. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of miR-140-5 P, VEGF, vascular endothelial growth factor receptor 2 (VEGFR2), PR, ER, AR, and ITGαvβ3 in rat endometrium. ResultsCompared with normal group, the estrous cycle of the rats in model group continued to be in the estrus interval and the estrous cycle lost regular changes. The endometrium was significantly thinner, the number of uterine glands and blood vessels were significantly reduced (P<0.01), and the pregnancy rate was significantly reduced. Compared with the model group, each drug group restored the regular estrous cycle to varying degrees, and the endometrial thickness and the number of blood vessels were significantly improved (P<0.01). The pregnancy rate of each drug group increased, and the effect of the cycle therapy group could reach the normal level. The results of molecular biology experiments showed that compared with the normal group, the levels of serum E2 and P in the model group were significantly decreased (P<0.01), the expression of VEGF, ER, PR and ITGαvβ3 protein was significantly decreased (P<0.05,P<0.01), the expression of AR protein was significantly increased (P<0.01), the expression of miR-140-5P and AR mRNA was significantly increased (P<0.01), and the expression of VEGF, VEGFR2, ER, PR and ITGαvβ3 mRNA was significantly decreased (P<0.01). Compared with model group, the serum E2 level in the Xiaoyaosan group was significantly increased (P<0.01).The levels of E2 and P in serum of rats in Sanzi Yangmo decoction group, cycle therapy group and aspirin group were significantly increased (P<0.01). The expression of AR protein in each drug group was significantly decreased (P<0.01). The expression of VEGF and ITGαvβ3 protein in Xiaoyaosan group was significantly increased (P<0.01). The expression of VEGF, ER and PR protein in Sanzi Yangmo decoction group was increased to varying degrees (P<0.05,P<0.01). The expression of VEGF, PR, ER and ITGαvβ3 protein in the cycle therapy group and the aspirin group increased to varying degrees (P<0.05,P<0.01). The expression of miR-140-5P and AR mRNA in each drug group was significantly decreased (P<0.01). The expression of VEGF, VEGFR2, ER, PR and ITGαvβ3 mRNA in each drug group increased to varying degrees (P<0.05,P<0.01). Compared with Xiaoyaosan group and Sanzi Yangmo decoction group, the expression of miR-140-5P, VEGFR2, ER, PR, AR and ITGαvβ3 mRNA in the cycle therapy group were significantly different (P<0.05,P<0.01). ConclusionThe kidney-tonifying and liver-regulating cyclical therapy may reduce the activity of miR-140-5P, target the upregulation of VEGF expression, mediate angiogenesis, and promote endometrial angiogenesis, thereby playing a synergistic role in improving endometrial receptivity in PCOS-induced infertility. Its efficacy in increasing pregnancy rates surpasses that of Xiaoyaosan or Sanzi Yangmo decoction used alone.


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