1.Analysis of clinical factors related to complete response after neoadjuvant chemoradiotherapy for locally advanced rectal cancer
Hui YANG ; Xiaofeng MU ; Linan SONG ; Wenjie NI ; Lei DING
Chinese Journal of Radiological Health 2026;35(1):6-11
Objective To explore the clinical factors influencing complete response in patients with locally advanced rectal cancer (LARC) after neoadjuvant chemoradiotherapy (nCRT). Methods Clinical data of LARC patients treated in the Department of Radiation Oncology at Beijing Shijitan Hospital between January 2013 and December 2024 were retrospectively collected. All patients received nCRT, after which surgery or a watch-and-wait approach was adopted based on treatment response. Univariable and multivariable logistic regression analyses were performed to identify prognostic factors influencing complete response. A clinical prediction model was constructed based on the multivariable analysis results, and its predictive performance was evaluated using the receiver operating characteristic curve. Results A total of 113 eligible patients were included. After nCRT, 19 patients (16.8%) achieved complete response, including 3 with clinical complete response and 16 with pathological complete response. Univariable analysis indicated that pretreatment clinical N stage, extramural venous invasion, carcinoembryonic antigen level, and neoadjuvant treatment regimen were associated with complete response after nCRT (P<0.05). Multivariable logistic regression analysis identified pretreatment extramural venous invasion, carcinoembryonic antigen level, and neoadjuvant treatment regimen as independent influencing factors for complete response (P<0.05). A prediction model incorporating these independent factors yielded an area under the receiver operating characteristic curve of 0.813 (95% confidence interval: 0.713-0.913), with a sensitivity of 89.5% and a specificity of 60.6%, demonstrating good predictive performance. Conclusion Pretreatment extramural venous invasion, carcinoembryonic antigen level, and neoadjuvant treatment regimen are independent factors influencing complete response after nCRT in LARC patients. The prediction model combining these factors may assist in evaluating treatment efficacy following nCRT in LARC patients.
2.Mechanism of Huayu jiedu formula in alleviating inflammatory injury in chronic kidney disease based on AIM2 pyroptosis pathway
Jinhuan XUE ; Ziwen WU ; Fan YANG ; Yunyun LOU ; Yingjun DING ; Yupeng XIAO ; Xianhui LIU ; Wenjie LIANG
China Pharmacy 2025;36(21):2638-2644
OBJECTIVE To explore the mechanism of Huayu jiedu formula in regulating inflammatory injury in chronic kidney disease (CKD). METHODS Fifty male SD rats were randomly divided into a sham surgery group (10 rats) and a modeling group (40 rats). The CKD model was replicated in the modeling group by unilateral ureteral obstruction surgery. After successful modeling, the rats were randomly divided into model group, esaxerenone group (positive control), and TCM low- and high-dose groups, with 10 rats in each group. The Esaxerenone group was given 1 mg/kg of esaxerenone, while the TCM low- and high-dose groups were given 13.7 and 27.4 g/kg of Huayu jiedu formula respectively, the sham surgery group and model group were given an equal volume of physiological saline, all groups were intervened continuously for 14 days. Hematoxylin eosin and Masson staining were used to observe the pathological changes in rat kidney tissue. Conventional biochemical methods were used to detect serum urea (SUr), serum creatinine (SCr), malondialdehyde (MDA), and superoxide dismutase (SOD) levels; enzyme-linked immunosorbent assay was used to detect the levels of serum interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α(TNF-α); immunohistochemistry and Western blot were used to detect the protein expression of peroxisome proliferator-activated receptor γ coactivator-1α(PGC-1α) , mitochondrial transcription factor A (TFAM), absent in melanoma 2(AIM2), caspase-1, gasdermin D (GSDMD), IL-1β and IL-18 in renal tissue; real-time fluorescence quantitative PCR was used to detect the mRNA expression of AIM2. RESULTS Compared with the sham surgery group, the renal tissue of the model group showed pathological changes such as glomerular deformation and destruction, severe tubular dilation, and increased deposition of blue fibrin; the levels of SUr, SCr, MDA, IL-1β, IL-6, TNF-α,the protein expression of AIM2, GSDMD, caspase-1, IL-1β, IL-18 , and the mRNA expression of AIM2 were significantly increased or up-regulated (P<0.01); the levels of SOD, the protein expression of PGC-1α, TFAM were significantly reduced or down-regulated (P<0.01). Compared with the model group, all treatment groups showed improvement in the above symptoms and most indicators in rats. CONCLUSIONS Huayu jiedu formula may improve renal function, alleviate renal inflammatory damage and pyroptosis, and exert renal protective effects by regulating the AIM2 pyroptosis pathway.
3.Whole-genome polymorphism of CD36 by third-generation sequencing technology
Jing LIU ; Xiuzhang XU ; Haoqiang DING ; Jing DENG ; Yangkai CHEN ; Wenjie XIA ; Xin YE
Chinese Journal of Blood Transfusion 2025;38(5):610-614
Objective: To analyze CD36 gene by PacBio Sequel Ⅱ the third-generation sequencing technology (TGS), including non-coding sequence, and to investigate the molecular mechanism of CD36 deficiency. Methods: Flow cytometry was performed in the southern Chinese population to detect the CD36 phenotype. Among them, 15 cases of CD36 type I deficiency, 15 cases of CD36 type Ⅱ deficiency, and 10 positive samples were selected. The TGS of the CD36 gene was performed and statistical analysis was conducted. Results: 40 samples (including 15 cases of type I deficiency, 15 cases of type Ⅱ deficiency, and 10 positive samples) were subjected by TGS of CD36 full-length sequences (except part of intron1). A total of 180 polymorphic loci were identified. Among them, 13 kinds were in the coding region, the rest were in non-coding region, with most mutations located in regulatory regions such as the 5′-UTR and 3′-UTR. Conclusion: The high polymorphism of CD36 non-coding regions, particularly in regulatory sequences, provides mechanistic insights into type Ⅱ CD36 deficiency.
4.Correlation between Muscle Tension,Clinical Characteristics, and Traditional Chinese Medicine Syndromes in Patients with Wilson Disease Based on Digital Muscle Function Assessment System Myoton PRO
Yulong YANG ; Wenming YANG ; Han WANG ; Xiang LI ; Taohua WEI ; Wenjie HAO ; Yue YANG ; Yufeng DING ; Yuqi SONG ; Wei HE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(15):147-154
ObjectiveThis paper aims to use the digital muscle function assessment system Myoton PRO to assess the correlation between muscle tension,clinical characteristics, and traditional Chinese medicine(TCM) syndromes in patients with hepatolenticular degeneration [also known as Wilson disease(WD)]. MethodsA total of 104 patients with WD accompanied by abnormal muscle tension(increased or decreased,hereinafter the same) who were hospitalized in the Brain Disease Centre of the First Affiliated Hospital of Anhui University of Chinese Medicine from April 2021 to November 2023 were selected,all of whom were subjected to TCM syndrome diagnosis and Myoton PRO for the measurement of F value of muscle tension,Goldstein, and UWDRS-N scales. The age of onset of the disease and disease duration were analyzed,and the differences and correlations of the above indexes in different TCM syndromes of WD were analyzed ResultsAmong the 104 patients with WD ,the phlegm and stasis syndrome was the most common(60 patients),followed by the damp-heat syndrome(33 patients),and the least common was the liver-kidney Yin deficiency syndrome(11 patients). The F value of the phlegm and stasis syndrome group was higher than that of the liver-kidney Yin deficiency syndrome group and the damp-heat syndrome group(P<0.01). The F value of the damp-heat syndrome group was higher than that of the liver-kidney Yin deficiency syndrome group(P<0.05),and the F value of the lower limbs of each group was higher than that of the upper limbs(P<0.01). Goldstein and UWDRS-N scores of the patients in the phlegm and stasis syndrome group were higher than those in the damp-heat syndrome group and the liver-kidney Yin deficiency syndrome group(P<0.05). There was no significant difference between the Goldstein and UWDRS-N scores of patients in the liver-kidney Yin deficiency syndrome group and the damp-heat syndrome group. Correlation analysis revealed that the age of onset and duration of the disease were positively correlated with the F values of the lower limbs(r=0.20,P<0.05,r=0.38,P<0.01)and had no significant correlation with those of the upper limbs. The F value levels of muscle tension of all limbs in the three groups of patients were positively correlated with the Goldstein and UWDRS-N scores(muscle tension of the upper limbs in the phlegm and stasis syndrome group,r=0.36,P<0.01,r=0.42,P<0.01. muscle tension of the lower limbs in the phlegm and stasis syndrome group,r=0.70,P<0.01,r=0.60,P<0.01. muscle tension of the upper limbs in the damp-heat syndrome group,r=0.64,P<0.01,r=0.53,P<0.01. muscle tension of the lower limbs in the damp-heat syndrome group,r=0.59,P<0.01,r=0.70,P<0.01. muscle tension of the upper limbs in the liver-kidney Yin deficiency syndrome group,r=0.70,P<0.01,r=0.74,P<0.01. muscle tension of the lower limbs in the liver-kidney Yin deficiency syndrome group,r=0.85,P<0.01,r=0.62,P<0.01).
5.Cuscutae Semen-Lycii Fructus Improves Spermatogenesis in Rat Model of Oligoasthenozoospermia by Inhibiting Oxidative Stress-induced Blood-testis Barrier Damage via Regulating SIRT1/Nrf2 Signaling Pathway
Wen DUAN ; Xiaojing ZHANG ; Wenjie DING ; Jianning JIN ; Guoqing CHU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):29-38
ObjectiveTo investigate the effect of the herb pair Cuscutae Semen-Lycii Fructus on oxidative stress-induced blood-testis barrier dysfunction and spermatogenesis in the rat model of oligoasthenozoospermia (OAS) and decipher the mechanism based on the silent information regulator 1 (SIRT1)/nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway. MethodsThirty-five male SD rats were randomized into a blank group (n=7) and a modeling group (n=28). The OAS model was established by gavage of hydrocortisone aqueous solution combined with single factor electrical stimulation. The modeled rats were randomly assigned into the following groups: model, Cuscutae Semen-Lycii Fructus granules (3.2 g·kg-1), Cuscutae Semen-Lycii Fructus total flavonoids (0.34 g·kg-1), and L-carnitine (0.38 g·kg-1), and treated for 4 weeks. The sperm quality of rats was assessed by an automatic sperm analyzer. The levels of superoxide dismutase (SOD), malondialdehyde (MAD), and glutathione peroxidase (GSH-Px) in the testicular tissue were determined by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining was employed to reveal the pathological changes in the testicular tissue and score the spermatogenic function. Transmission electron microscopy was employed to observe the ultrastructural changes of Sertoli cells. Western blot and Real-time PCR were employed to determine the protein and mRNA levels, respectively, of SIRT1, Nrf2, Occludin, zonula occludens-1 (ZO-1), connexin 43 (CX43), and β-catenin. ResultsCompared with the blank group, the model group showed decreased total sperm count and motility (P<0.05, P<0.01), obvious damage in the testicular tissue and blood-testis barrier structure, reduced score of spermatogenic function (P<0.01), declined levels of GSH-Px and SOD in the testicular tissue (P<0.05), elevated level of MDA, and down-regulated protein levels of SIRT1, Nrf2, ZO-1, CX43, β-catenin, and occludin (P<0.05, P<0.01) and mRNA levels of SIRT1, Nrf2, ZO-1, CX43, and β-catenin in the testicular tissue (P<0.05, P<0.01). After treatment, the testicular tissue, blood-testis barrier structure, and score of spermatogenic function (P<0.01) were improved in the Cuscutae Semen-Lycii Fructus granules group, Cuscutae Semen-Lycii Fructus total flavonoids group, and L-carnitine group. Compared with the model group, the treatment groups presented lowered levels of GSH-Px and SOD (P<0.05, P<0.01), and the Cuscutae Semen-Lycii Fructus granule group showed a decline in MDA level. The protein and mRNA levels of SIRT1, Nrf2, ZO-1, CX43, β-catenin, and occludin were up-regulated in the Cuscutae Semen-Lycii Fructus granules group and total flavonoids group (P<0.05, P<0.01). ConclusionThe herb pair Cuscutae Semen-Lycii Fructus can regulate the SIRT1/Nrf2 pathway to inhibit oxidative stress and alleviate the blood-testis barrier damage, thereby improving the spermatogenic function in the rat model of OAS. Total flavonoids may be the material basis for the therapeutic effect of Cuscutae Semen-Lycii Fructus.
6.Cuscutae Semen-Lycii Fructus Improves Spermatogenesis in Rat Model of Oligoasthenozoospermia by Inhibiting Oxidative Stress-induced Blood-testis Barrier Damage via Regulating SIRT1/Nrf2 Signaling Pathway
Wen DUAN ; Xiaojing ZHANG ; Wenjie DING ; Jianning JIN ; Guoqing CHU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):29-38
ObjectiveTo investigate the effect of the herb pair Cuscutae Semen-Lycii Fructus on oxidative stress-induced blood-testis barrier dysfunction and spermatogenesis in the rat model of oligoasthenozoospermia (OAS) and decipher the mechanism based on the silent information regulator 1 (SIRT1)/nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway. MethodsThirty-five male SD rats were randomized into a blank group (n=7) and a modeling group (n=28). The OAS model was established by gavage of hydrocortisone aqueous solution combined with single factor electrical stimulation. The modeled rats were randomly assigned into the following groups: model, Cuscutae Semen-Lycii Fructus granules (3.2 g·kg-1), Cuscutae Semen-Lycii Fructus total flavonoids (0.34 g·kg-1), and L-carnitine (0.38 g·kg-1), and treated for 4 weeks. The sperm quality of rats was assessed by an automatic sperm analyzer. The levels of superoxide dismutase (SOD), malondialdehyde (MAD), and glutathione peroxidase (GSH-Px) in the testicular tissue were determined by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining was employed to reveal the pathological changes in the testicular tissue and score the spermatogenic function. Transmission electron microscopy was employed to observe the ultrastructural changes of Sertoli cells. Western blot and Real-time PCR were employed to determine the protein and mRNA levels, respectively, of SIRT1, Nrf2, Occludin, zonula occludens-1 (ZO-1), connexin 43 (CX43), and β-catenin. ResultsCompared with the blank group, the model group showed decreased total sperm count and motility (P<0.05, P<0.01), obvious damage in the testicular tissue and blood-testis barrier structure, reduced score of spermatogenic function (P<0.01), declined levels of GSH-Px and SOD in the testicular tissue (P<0.05), elevated level of MDA, and down-regulated protein levels of SIRT1, Nrf2, ZO-1, CX43, β-catenin, and occludin (P<0.05, P<0.01) and mRNA levels of SIRT1, Nrf2, ZO-1, CX43, and β-catenin in the testicular tissue (P<0.05, P<0.01). After treatment, the testicular tissue, blood-testis barrier structure, and score of spermatogenic function (P<0.01) were improved in the Cuscutae Semen-Lycii Fructus granules group, Cuscutae Semen-Lycii Fructus total flavonoids group, and L-carnitine group. Compared with the model group, the treatment groups presented lowered levels of GSH-Px and SOD (P<0.05, P<0.01), and the Cuscutae Semen-Lycii Fructus granule group showed a decline in MDA level. The protein and mRNA levels of SIRT1, Nrf2, ZO-1, CX43, β-catenin, and occludin were up-regulated in the Cuscutae Semen-Lycii Fructus granules group and total flavonoids group (P<0.05, P<0.01). ConclusionThe herb pair Cuscutae Semen-Lycii Fructus can regulate the SIRT1/Nrf2 pathway to inhibit oxidative stress and alleviate the blood-testis barrier damage, thereby improving the spermatogenic function in the rat model of OAS. Total flavonoids may be the material basis for the therapeutic effect of Cuscutae Semen-Lycii Fructus.
7.In vitro expression analysis of the ITGB3 c.598G/A mutation and its association with FNAIT
Haoqiang DING ; Xin YE ; Xiuzhang XU ; Wenjie XIA ; Jing DENG ; Jing LIU ; Yangkai CHEN ; Dawei CHEN ; Yaori XU
Chinese Journal of Blood Transfusion 2025;38(7):873-878
Objective: To explore the role of the c.598G>A mutation of the ITGB3 gene in the occurrence of fetal and neonatal alloimmune thrombocytopenia (FNAIT) through its expression in vitro. Methods: The platelet antibodies in the sera of the affected neonate and her mother were detected using commercial enzyme-linked immunosorbent assay (ELISA), solid-phase agglutination, flow cytometry and the gold standard monoclonal antibody-specific immobilization of platelet antigens (MAIPA). The common human platelet antigen (HPA) genotypes of the neonate and her parents were obtained using the HPA-SSP method. The presence of mutations was analyzed by sequencing the exons of the ITGB3 and ITGA2B genes. The target gene of ITGB3 was obtained by PCR amplification using the existing human platelet cDNA. The wild-type ITGB3 eukaryotic expression vector was constructed by TA cloning technology. The 598G>A mutant ITGB3 eukaryotic expression vector was obtained by point mutation, and the plasmid DNA was co-transfected with that of ITGA2B (αⅡb) into HEK293 cells. The transfected cells stably expressing GP Ⅱb/Ⅲa were screened and obtained. The expression of GP Ⅱb/Ⅲa in 598G>A mutant transfected cells and the presence of antibodies against this mutation in the serum of mother were detected by flow cytometry and MAIPA. Results: Antibodies against HLA-class Ⅰ and GP Ⅱb/Ⅲa glycoproteins were detected in the serum of the neonate's mother, and subsequent HLA antibody-specific testing confirmed the presence of antibodies against HLA-B
57∶01 and A
02∶05. ITGB3 sequencing showed that the neonate and her father carried the c.598G>A point mutation, which results in the change of glutamate to lysine at position 200. Antibodies against GP Ⅱb/Ⅲa glycoproteins were not detected using constructed c.598G>A mutant transfected cells reacted with the maternal serum. Conclusion: The in vitro expression and analysis of the ITGB3 c.598G>A mutation did not support a role for this mutation in the pathogenesis of FNAIT. The establishment of this method facilitates the discovery of new platelet low-frequency antigens, and provides a theoretical foundation for the detection of antibodies against platelet antigens associated with patients with adverse pregnancy and childbirth histories.
8.In vitro expression analysis of the ITGB3 c.598G/A mutation and its association with FNAIT
Haoqiang DING ; Xin YE ; Xiuzhang XU ; Wenjie XIA ; Jing DENG ; Jing LIU ; Yangkai CHEN ; Dawei CHEN ; Yaori XU
Chinese Journal of Blood Transfusion 2025;38(7):873-878
Objective: To explore the role of the c.598G>A mutation of the ITGB3 gene in the occurrence of fetal and neonatal alloimmune thrombocytopenia (FNAIT) through its expression in vitro. Methods: The platelet antibodies in the sera of the affected neonate and her mother were detected using commercial enzyme-linked immunosorbent assay (ELISA), solid-phase agglutination, flow cytometry and the gold standard monoclonal antibody-specific immobilization of platelet antigens (MAIPA). The common human platelet antigen (HPA) genotypes of the neonate and her parents were obtained using the HPA-SSP method. The presence of mutations was analyzed by sequencing the exons of the ITGB3 and ITGA2B genes. The target gene of ITGB3 was obtained by PCR amplification using the existing human platelet cDNA. The wild-type ITGB3 eukaryotic expression vector was constructed by TA cloning technology. The 598G>A mutant ITGB3 eukaryotic expression vector was obtained by point mutation, and the plasmid DNA was co-transfected with that of ITGA2B (αⅡb) into HEK293 cells. The transfected cells stably expressing GP Ⅱb/Ⅲa were screened and obtained. The expression of GP Ⅱb/Ⅲa in 598G>A mutant transfected cells and the presence of antibodies against this mutation in the serum of mother were detected by flow cytometry and MAIPA. Results: Antibodies against HLA-class Ⅰ and GP Ⅱb/Ⅲa glycoproteins were detected in the serum of the neonate's mother, and subsequent HLA antibody-specific testing confirmed the presence of antibodies against HLA-B
57∶01 and A
02∶05. ITGB3 sequencing showed that the neonate and her father carried the c.598G>A point mutation, which results in the change of glutamate to lysine at position 200. Antibodies against GP Ⅱb/Ⅲa glycoproteins were not detected using constructed c.598G>A mutant transfected cells reacted with the maternal serum. Conclusion: The in vitro expression and analysis of the ITGB3 c.598G>A mutation did not support a role for this mutation in the pathogenesis of FNAIT. The establishment of this method facilitates the discovery of new platelet low-frequency antigens, and provides a theoretical foundation for the detection of antibodies against platelet antigens associated with patients with adverse pregnancy and childbirth histories.
9.Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury (version 2025)
Aijun XU ; Shuixia LI ; Bo CHEN ; Mengyuan YE ; Lejiao LANG ; Ning NING ; Lin ZHANG ; Changqing LIU ; Zhonglan CHEN ; Weihu MA ; Weishi LI ; Xiaoning WANG ; Dongmei BIAN ; Jiancheng ZENG ; Xin WANG ; Yuan GAO ; Yaping CHEN ; Jiali CHEN ; Yun HAN ; Xiuting LI ; Yang ZHOU ; Xiaojing SU ; Qiong ZHANG ; Tianwen HUANG ; Ping ZHANG ; Hua LIN ; Xingling XIAO ; Ruifeng XU ; Fanghui DONG ; Bing HAN ; Luo FAN ; Yanling PEI ; Suyun LI ; Xiaoju TAN ; Rongchen GUO ; Yefang ZOU ; Xiaoyun HAN ; Junqin DING ; Yi WANG ; Shuhua DENG ; Jinli GUO ; Yinhua LIANG ; Yuan CEN ; Xiaoqin LIU ; Junru CHEN ; Haiyang YU ; Lunlan LI ; Ying REN ; Yunxia LI ; Jianli LU ; Ying YING ; Lan WEI ; Yin WANG ; Qinhong XU ; Yanqin ZHANG ; Yang LYU ; Shijun ZHANG ; Sui WENJIE ; Sanlian HU ; Shuhong YANG ; Guoqing LI ; Jingjing AN ; Baorong HE ; Leling FENG
Chinese Journal of Trauma 2025;41(6):530-541
Paraplegia caused by spinal cord injury is a serious neurological complication, for which surgery is currently the main treatment method. Due to different surgical approaches, patients are usually expected to maintain a passive prone position for a long time or switch between the supine and prone positions. Affected by multiple factors such as neurogenic sensory disorders, pathological changes in muscle tone and operative duration, the risk of intraoperative acquired pressure injury (IAPI) is significantly increased. Current clinical prevention strategies for IAPI in these patients predominantly focus on localized pressure relief during positioning, lacking systematic, standardized comprehensive prevention protocols or evidence-based guidelines. To address it, Department of Nursing, Orthopedics Branch, China International Exchange and Promotive Association for Medical and Health Care, Spinal Trauma Professional Committee, Orthopedics Branch, Chinese Medical Doctor Association, Nursing Group of Spine and Spinal Cord Professional Committee of Chinese Association of Rehabilitation Medicine organized experts in relevant fields to formulate Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury ( version 2025), based on evidence-based medical evidence and latest research results and clinical practice at home and abroad. Eleven recommendations were put forward from the aspects of preoperative risk assessment, intraoperative prevention strategies, postoperative handover and monitoring, and supportive mechanisms for IAPI prevention, aiming to standardize the prevention measures and management strategies of IAPI in paraplegic patients with spinal cord injury and accelerate the recovery of patients and improve the therapeutic effect.
10.Head electroacupuncture alleviates central hyperalgesia in rats with knee osteoarthritis
Bohan HAI ; Hui LI ; Wenjie LIU ; Han WANG ; Chenchen WU ; Cuiping GUO ; Xinju DING ; Wenbiao WANG
Chinese Journal of Physical Medicine and Rehabilitation 2025;47(7):585-594
Objective:To explore the effect of scalp electroacupuncture (EA) on central pain sensitization in rats with knee osteoarthritis (KOA).Methods:Thirty-two 8-week-old female Sprague-Dawley rats were randomly divided into a blank control group, a model group, an electroacupuncture (EA) group and a sham EA group, each of 8. All of the rats except those in the control group had KOA induced through intra-articular monosodium iodoacetate injections in the right knee. Two weeks later the EA group rats began receiving daily head EA sessions 6 days/week for 2 weeks. The sham EA group received identical but non-therapeutic stimulation. The blank control and model groups received no EA intervention. Before the modelling and 3, 7, 14, 21 and 28 days later, all of the rats completed bipedal balance pain tests and mechanical allodynia evaluations. After the testing on day 28, all of the rats were euthanized for molecular analyses. Western blotting and immunohistochemistry were performed to examine protein expression of brain-derived neurotrophic factor (BDNF), tropomyosin receptor kinase B (TrkB), and cannabinoid receptor type 1 (CB1R) in both the periaqueductal gray (PAG) matter and spinal dorsal horns (SDHs). Serum levels of substance P (SP) and 5-hydroxytryptamine (5-HT) were also quantified using enzyme-linked immunosorbent assays.Results:Three days after successful modeling, the average weight-bearing capacity of the right hind limb in the model, sham EA and EA groups was significantly lower than that of the blank controls. It reached its lowest level on the 14th day after modeling. Concurrently, the pain responses in those three groups were significantly higher than among the controls, also peaking on the 14th day after modeling. After two weeks of electroacupuncture, the electroacupuncture group showed significant improvement in both right hind limb weight-bearing capacity and pain response compared to the model group. Meanwhile, the levels of BDNF and TrkB protein in the periaqueductal gray matter were significantly higher in the model group than among the blank controls, while the electroacupuncture group exhibited significantly reduced expression of BDNF and TrkB proteins compared to the model group, along with significantly increased CB1R protein expression. The model group showed significantly elevated expression of both BDNF and TrkB proteins in the spinal dorsal horn compared to the blank control group, while there were significant differences between the EA and model groups in the expression of BDNF, TrkB and CB1R proteins. Immunohistochemical analysis on day 28 revealed that the EA group had significantly fewer BDNF- and TrkB-positive cells in the PAG compared to the model group, with significantly more CB1R-positive cells. In the SDH, the model group exhibited significantly increased numbers of BDNF- and TrkB-positive cells compared to the blank control group, whereas significant differences were found between the EA and blank control groups in the numbers of BDNF-, TrkB- and CB1R-positive cells. Serum analysis on day 28 demonstrated that substance P and 5-hydroxytryptamine levels in the model, sham EA and EA groups were significantly higher than in the blank control group, on average. However, no significant differences were observed in serum SP and 5-HT levels between the EA and model groups.Conclusions:Scalp EA significantly alleviates central pain sensitization in KOA, at least in rats, potentially by suppressing BDNF and TrkB expression while upregulating CB1R expression in the PAG matter and the SDH.

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