1.Cognitive Characteristics and Empirical Research Paths of Tacit Knowledge in Tongue Inspection of Traditional Chinese Medicine
Min LAI ; Chunhua JIA ; Yuxia JIN ; Ying HU ; Yuan DENG
Journal of Traditional Chinese Medicine 2026;67(13):1353-1358
Tacit knowledge of tongue inspection is content that affects diagnostic results but is difficult to be explicit. Its cognitive characteristics are rooted in xiang thinking and embodiment. It constructs multidimensional associations between tongue manifestations and disease mechanisms through imagery mapping, and it relies on the practitioner's bodily perception, clinical context, and long-term practice for continuous internalization. In response to the neglect of tacit transmission and individual cognitive processes in current traditional Chinese medicine (TCM) tacit knowledge research, an empirical research framework integrating cognitive neuroscience is proposed. Specifically, event-related potential (ERP) and eye tracking (ET) technologies can be used to capture the brain electrical activity and eye movement trajectories of experts and novices in different tongue diagnosis cognitive tasks. By integrating multimodal cognitive data such as behaviorism, ERP components, and ET indicators, the implicit cognitive advantages of experts in the perception, mapping, and decision-making stages of tongue diagnosis are presented, and the possibility of achieving tacit knowledge transformation through visual cognitive transfer training is explored. Exploring empirical pathways to uncover the cognitive mechanisms of tongue diagnosis tacit knowledge is of great significance for the inheritance of traditional Chinese medicine.
2.Fangji Fulingtang Attenuates Myocardial Fibrosis by Regulating Mitochondrial Function Through AMPK/PGC-1α/MFN2-PKM2 Signaling Pathway
Xuqin DU ; Yixuan LI ; Yuxia JIN ; Hongding LI ; Ruogu YANG ; Lipeng SHI ; Yi REN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):1-10
ObjectiveTo investigate the effects and mechanisms of Fangji Fulingtang (FFD) on mitochondrial function in the mouse model of myocardial fibrosis (MF). MethodsSixty SPF-grade male C57BL/6J mice were randomly allocated into six groups (n=10 per group): control, model, low-dose, medium-dose, and high-dose (3.315, 6.63, and 13.26 g·kg-1, respectively) FFD, and captopril (20 mg·kg-1). MF was induced by subcutaneous injection of isoproterenol (10 mg·kg-1·d-1) in other groups except the control group for 14 consecutive days, with simultaneous gavage of corresponding drugs. Left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were measured by echocardiography. Serum levels of creatine kinase-MB (CK-MB), cardiac troponin I (cTnI), N-terminal pro-brain natriuretic peptide (NT-pro BNP), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) were determined by enzyme-linked immunosorbent assay (ELISA). Malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) levels were measured by biochemical assays. Reactive oxygen species (ROS) were detected by dihydroethidium (DHE) fluorescence staining. Hematoxylin-eosin (HE), Masson's trichrome, and wheat germ agglutinin (WGA) staining were performed to evaluate myocardial structure and fibrosis. Mitochondrial ultrastructure and function were assessed by transmission electron microscopy, adenosine triphosphate (ATP) colorimetric assay, and JC-1 fluorescence staining. The protein levels of phosphorylated adenosine monophosphate-activated protein kinase α subunit (p-AMPKα), AMPKα, peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α), mitofusin 2 (MFN2), pyruvate kinase M2 isoform (PKM2), lactate dehydrogenase A (LDHA), and hypoxia-inducible factor 1 alpha (HIF-1α) were analyzed by Western blot. ResultsCompared with the control group, the model group exhibited decreased LVEF and LVFS, increased heart weight index and heart weight-to-tibia length ratio (P<0.01), elevated levels of myocardial injury markers (CK-MB, cTnI, and NT-pro BNP), inflammatory cytokines (TNF-α, IL-1β, and IL-6), and MDA, along with reduced SOD and GSH levels (P<0.01). Enhanced interstitial collagen deposition and cardiomyocyte hypertrophy were observed in the model group (P<0.01). Transmission electron microscopy and JC-1 staining revealed mitochondrial swelling, crista disruption, decreased ATP content, and reduced red/green fluorescence ratio in the model group (P<0.01). Western blot analysis demonstrated downregulation of p-AMPKα, PGC-1α, and MFN2 and upregulation of PKM2, LDHA, and HIF-1α in the model group (P<0.01). Compared with the model group, treatment with FFD or captopril improved LVEF and LVFS, reduced heart weight index and heart weight-to-tibia length ratio (P<0.05, P<0.01), lowered the serum levels of CK-MB, cTnI, NT-pro BNP, TNF-α, IL-1β, IL-6, and MDA, increased the SOD and GSH levels (P<0.05, P<0.01), attenuated the myocardial fibrosis and cardiomyocyte hypertrophy (P<0.01), and restored the mitochondrial ultrastructure. The medium and high-dose FFD groups as well as the captopril group showed increased ATP production and red/green fluorescence ratio (P<0.05, P<0.01). Furthermore, FFD upregulated the expression of p-AMPKα and PGC-1α while downregulating the expression of LDHA and HIF-1α (P<0.05, P<0.01). ConclusionFFD activates the AMPK/PGC-1α/MFN2 signaling pathway and inhibits the PKM2/LDHA/HIF-1α axis to restore mitochondrial function and energy metabolic homeostasis, thereby attenuating isoproterenol-induced myocardial fibrosis.
3.Fangji Fulingtang Attenuates Myocardial Fibrosis by Regulating Mitochondrial Function Through AMPK/PGC-1α/MFN2-PKM2 Signaling Pathway
Xuqin DU ; Yixuan LI ; Yuxia JIN ; Hongding LI ; Ruogu YANG ; Lipeng SHI ; Yi REN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):1-10
ObjectiveTo investigate the effects and mechanisms of Fangji Fulingtang (FFD) on mitochondrial function in the mouse model of myocardial fibrosis (MF). MethodsSixty SPF-grade male C57BL/6J mice were randomly allocated into six groups (n=10 per group): control, model, low-dose, medium-dose, and high-dose (3.315, 6.63, and 13.26 g·kg-1, respectively) FFD, and captopril (20 mg·kg-1). MF was induced by subcutaneous injection of isoproterenol (10 mg·kg-1·d-1) in other groups except the control group for 14 consecutive days, with simultaneous gavage of corresponding drugs. Left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were measured by echocardiography. Serum levels of creatine kinase-MB (CK-MB), cardiac troponin I (cTnI), N-terminal pro-brain natriuretic peptide (NT-pro BNP), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) were determined by enzyme-linked immunosorbent assay (ELISA). Malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) levels were measured by biochemical assays. Reactive oxygen species (ROS) were detected by dihydroethidium (DHE) fluorescence staining. Hematoxylin-eosin (HE), Masson's trichrome, and wheat germ agglutinin (WGA) staining were performed to evaluate myocardial structure and fibrosis. Mitochondrial ultrastructure and function were assessed by transmission electron microscopy, adenosine triphosphate (ATP) colorimetric assay, and JC-1 fluorescence staining. The protein levels of phosphorylated adenosine monophosphate-activated protein kinase α subunit (p-AMPKα), AMPKα, peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α), mitofusin 2 (MFN2), pyruvate kinase M2 isoform (PKM2), lactate dehydrogenase A (LDHA), and hypoxia-inducible factor 1 alpha (HIF-1α) were analyzed by Western blot. ResultsCompared with the control group, the model group exhibited decreased LVEF and LVFS, increased heart weight index and heart weight-to-tibia length ratio (P<0.01), elevated levels of myocardial injury markers (CK-MB, cTnI, and NT-pro BNP), inflammatory cytokines (TNF-α, IL-1β, and IL-6), and MDA, along with reduced SOD and GSH levels (P<0.01). Enhanced interstitial collagen deposition and cardiomyocyte hypertrophy were observed in the model group (P<0.01). Transmission electron microscopy and JC-1 staining revealed mitochondrial swelling, crista disruption, decreased ATP content, and reduced red/green fluorescence ratio in the model group (P<0.01). Western blot analysis demonstrated downregulation of p-AMPKα, PGC-1α, and MFN2 and upregulation of PKM2, LDHA, and HIF-1α in the model group (P<0.01). Compared with the model group, treatment with FFD or captopril improved LVEF and LVFS, reduced heart weight index and heart weight-to-tibia length ratio (P<0.05, P<0.01), lowered the serum levels of CK-MB, cTnI, NT-pro BNP, TNF-α, IL-1β, IL-6, and MDA, increased the SOD and GSH levels (P<0.05, P<0.01), attenuated the myocardial fibrosis and cardiomyocyte hypertrophy (P<0.01), and restored the mitochondrial ultrastructure. The medium and high-dose FFD groups as well as the captopril group showed increased ATP production and red/green fluorescence ratio (P<0.05, P<0.01). Furthermore, FFD upregulated the expression of p-AMPKα and PGC-1α while downregulating the expression of LDHA and HIF-1α (P<0.05, P<0.01). ConclusionFFD activates the AMPK/PGC-1α/MFN2 signaling pathway and inhibits the PKM2/LDHA/HIF-1α axis to restore mitochondrial function and energy metabolic homeostasis, thereby attenuating isoproterenol-induced myocardial fibrosis.
4.Antimicrobial resistance surveillance in the bacterial strains isolated from pediatric intensive care units in China:results from 2020 to 2022
Jing LIU ; Huiyuan YAN ; Gangfeng YAN ; Guoping LU ; Pan FU ; Chuanqing WANG ; Danqun JIN ; Wenjia TONG ; Chenyu ZHANG ; Jianli CHEN ; Yi LIN ; Jia LEI ; Yibing CHENG ; Qunqun ZHANG ; Kaijie GAO ; Yuanyuan CHEN ; Shufang XIAO ; Juan HE ; Li JIANG ; Huimin XU ; Yuxia LI ; Hanghai DING ; Hehe CHEN ; Yao ZHENG ; Qunying CHEN ; Ying WANG ; Hong REN ; Chenmei ZHANG ; Zhenjie CHEN ; Mingming ZHOU ; Yucai ZHANG ; Yiping ZHOU ; Zhenjiang BAI ; Saihu HUANG ; Lili HUANG ; Weiguo YANG ; Weike MA ; Qing MENG ; Pengwei ZHU ; Yong LI ; Yan XU ; Yi WANG ; Yanqiang DU ; Huijun CAI ; Bizhen ZHU ; Huixuan SHI ; Shaoxian HONG ; Yukun HUANG ; Meilian HUANG
Chinese Journal of Infection and Chemotherapy 2025;25(3):303-311
Objective This study aimed to investigate the antimicrobial resistance profiles of bacterial strains isolated from pediatric intensive care units(PICU)in China for better antimicrobial therapy.Methods Clinical isolates were collected from 17 institutions,including tertiary care children's hospitals and pediatric department of tertiary general hospitals in China from January 1,2020 to December 31,2022.Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated systems.Results were interpreted according to the breakpoints released by the Clinical and Laboratory Standards Institute(CLSI)in 2020.Results A total of 10 688 isolates were collected,including gram-positive organisms(39.2%)and gram-negative organisms(60.8%).The top three organisms were S.aureus(13.6%,1 453/10 688),A.baumannii(10.0%,1 067/10 688),and coagulase-negative Staphylococcus(9.9%,1 058/10 688).Multi-drug resistant organisms(MDROs)were very common in children.The prevalence of methicillin-resistant Staphylococcus aureus(MRSA),carbapenem-resistant Enterobacterales(CRE),carbapenem-resistant E.coli,carbapenem-resistant K.pneumoniae(CRKP),carbapenem-resistant A.baumannii(CRAB),and carbapenem-resistant P.aeruginosa(CRPA)was 41.1%,19.4%,8.8%,30.9%,67.4%,and 28.8%,respectively.Overall,more than 50%of Enterobacteriales isolates were resistant to cephalosporins,while nearly 25%of Enterobacteriales isolates were resistant to carbapenems.MDROs were highly resistant to commonly used antibiotics.More than 80%of CRE and CRAB strains were resistant to all beta-lactam antibiotics.CRE and CRAB showed low resistance rates to tigecycline and polymyxin.CRPA showed lower resistance rates to piperacillin,beta-lactamase inhibitor combinations than the resistance rates to third and fourth generation cephalosporins.All of the Staphylococcus and Enterococcus isolates were susceptible to vancomycin and tigecycline.None of PRSP strains isolated from meningitis and nonmeningitis samples were resistant to rifampicin,vancomycin,or linezolid.The prevalence of β-lactamase-negative ampicillin-resistant(BLNAR)strains was 43.3%in Haemophilus influenzae.Conclusions MDROs were prevalent in PICU.It is necessary to establish an effective multidisciplinary team(MDT)to control the antimicrobial resistance.
5.Antimicrobial resistance surveillance in the bacterial strains isolated from pediatric intensive care units in China:results from 2020 to 2022
Jing LIU ; Huiyuan YAN ; Gangfeng YAN ; Guoping LU ; Pan FU ; Chuanqing WANG ; Danqun JIN ; Wenjia TONG ; Chenyu ZHANG ; Jianli CHEN ; Yi LIN ; Jia LEI ; Yibing CHENG ; Qunqun ZHANG ; Kaijie GAO ; Yuanyuan CHEN ; Shufang XIAO ; Juan HE ; Li JIANG ; Huimin XU ; Yuxia LI ; Hanghai DING ; Hehe CHEN ; Yao ZHENG ; Qunying CHEN ; Ying WANG ; Hong REN ; Chenmei ZHANG ; Zhenjie CHEN ; Mingming ZHOU ; Yucai ZHANG ; Yiping ZHOU ; Zhenjiang BAI ; Saihu HUANG ; Lili HUANG ; Weiguo YANG ; Weike MA ; Qing MENG ; Pengwei ZHU ; Yong LI ; Yan XU ; Yi WANG ; Yanqiang DU ; Huijun CAI ; Bizhen ZHU ; Huixuan SHI ; Shaoxian HONG ; Yukun HUANG ; Meilian HUANG
Chinese Journal of Infection and Chemotherapy 2025;25(3):303-311
Objective This study aimed to investigate the antimicrobial resistance profiles of bacterial strains isolated from pediatric intensive care units(PICU)in China for better antimicrobial therapy.Methods Clinical isolates were collected from 17 institutions,including tertiary care children's hospitals and pediatric department of tertiary general hospitals in China from January 1,2020 to December 31,2022.Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated systems.Results were interpreted according to the breakpoints released by the Clinical and Laboratory Standards Institute(CLSI)in 2020.Results A total of 10 688 isolates were collected,including gram-positive organisms(39.2%)and gram-negative organisms(60.8%).The top three organisms were S.aureus(13.6%,1 453/10 688),A.baumannii(10.0%,1 067/10 688),and coagulase-negative Staphylococcus(9.9%,1 058/10 688).Multi-drug resistant organisms(MDROs)were very common in children.The prevalence of methicillin-resistant Staphylococcus aureus(MRSA),carbapenem-resistant Enterobacterales(CRE),carbapenem-resistant E.coli,carbapenem-resistant K.pneumoniae(CRKP),carbapenem-resistant A.baumannii(CRAB),and carbapenem-resistant P.aeruginosa(CRPA)was 41.1%,19.4%,8.8%,30.9%,67.4%,and 28.8%,respectively.Overall,more than 50%of Enterobacteriales isolates were resistant to cephalosporins,while nearly 25%of Enterobacteriales isolates were resistant to carbapenems.MDROs were highly resistant to commonly used antibiotics.More than 80%of CRE and CRAB strains were resistant to all beta-lactam antibiotics.CRE and CRAB showed low resistance rates to tigecycline and polymyxin.CRPA showed lower resistance rates to piperacillin,beta-lactamase inhibitor combinations than the resistance rates to third and fourth generation cephalosporins.All of the Staphylococcus and Enterococcus isolates were susceptible to vancomycin and tigecycline.None of PRSP strains isolated from meningitis and nonmeningitis samples were resistant to rifampicin,vancomycin,or linezolid.The prevalence of β-lactamase-negative ampicillin-resistant(BLNAR)strains was 43.3%in Haemophilus influenzae.Conclusions MDROs were prevalent in PICU.It is necessary to establish an effective multidisciplinary team(MDT)to control the antimicrobial resistance.
6.Effect of Shenshu Fujian Decoction on PDGF/NKD2/Wnt Signaling Pathway in Rats with Chronic Renal Failure
Peng DENG ; Xuekuan HUANG ; Hongyu LUO ; Yuxia JIN ; Dandan WANG ; Xin CHEN ; Shuxian YANG ; Honglin WANG ; Munan WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(15):79-86
ObjectiveTo observe the effect of Shenshu Fujian decoction on platelet-derived growth factor (PDGF)/naked cuticle homolog 2 (NKD2) /Wnt signaling pathway in rats with chronic renal failure (CRF). MethodsSixty male SD rats were randomly divided into normal group, model group, Niaoduqing group (5 g·kg-1), low-dose Shenshu Fujian decoction group (5.5 g·kg-1), medium-dose Shenshu Fujian decoction group (11 g·kg-1), and high-dose Shenshu Fujian decoction group (22 g·kg-1), with 10 rats in each group. A CRF rat model was established by feeding a 0.5% adenine diet for 21 days. After successful modeling, intragastric administration was given once daily for 28 consecutive days. After treatment, the renal morphology of rats was observed. Serum creatinine (SCr) and blood urea nitrogen (BUN) levels were detected. Hematoxylin-eosin (HE) staining and Masson staining were used to detect renal histopathological changes, and collagen volume fraction (CVF) was calculated. Serum levels of inflammatory markers interleukin (IL)-1β and IL-6 were measured using enzyme-linked immunosorbent assay (ELISA). The expressions of fibronectin 1 (FN1), type Ⅰ collagen (ColⅠ), α-smooth muscle actin (α-SMA), platelet-derived growth factor receptor-β (PDGFR-β), NKD2, dishevelled protein 2 (DVL2) and β-catenin in renal tissue were detected by immunohistochemistry and Western blot. ResultsCompared with the normal group, the model group showed significant renal pathological changes, a markedly increased kidney weight/body weight ratio (P<0.01), significantly elevated CVF (P<0.01), and notably increased serum levels of SCr, BUN, IL-1β, and IL-6 (P<0.01). Expression levels of FN1, ColⅠ, α-SMA, PDGFR-β, NKD2, DVL2, and β-catenin in renal tissue were also significantly increased (P<0.01). Compared with the model group, all treatment groups showed significantly decreased kidney weight/body weight ratios and CVF (P<0.01), as well as markedly decreased serum SCr, BUN, IL-1β, and IL-6 levels. Protein expression levels of FN1, ColⅠ, α-SMA, PDGFR-β, NKD2, DVL2, and β-catenin in renal tissue were decreased, with more pronounced effects observed in the Niaoduqing, medium-dose, and high-dose Shenshu Fujian decoction groups (P<0.05, P<0.01). ConclusionShenshu Fujian decoction improves renal function, reduces inflammation, and reverses renal fibrosis in CRF rats, possibly by downregulating the expression of PDGF/NKD2/Wnt signaling pathway-related proteins.
7.The academic thought of"reinforcing healthy Qi and unblocking regulation"in JIN Yicheng's Shanghai-style pediatric Tuina school and its clinical application
Qian WANG ; Yijing SHEN ; Jiqiong LU ; Yuxia CHEN ; Yicheng JIN ; Jue HONG
Journal of Acupuncture and Tuina Science 2025;23(4):367-376
The Shanghai-style pediatric Tuina(Chinese therapeutic massage)school,a renowned academic school of pediatric Tuina in China,was founded by Professor JIN Yicheng,a mentor of the National Senior Traditional Chinese Medicine(TCM)Experts'Clinical Experience Inheritance Class and a distinguished TCM practitioner in Shanghai.This academic school has now been perpetuated through four generations.Prof.JIN Yicheng,a pioneering leader in modern China's pediatric Tuina,has dedicated sixty years to medical practice with unwavering benevolence and adherence to"principled innovation".While delving into traditional and ancient teachings,he has also embraced contemporary advancements.Building upon the essence of traditional pediatric Tuina,he integrated distinctive techniques from various Tuina schools,including the Yi Zhi Chan Tuina school,rolling manipulation Tuina school,and Neigong Tuina school.He also assimilated the quintessence of historical pediatric Tuina literature,the experience of modern Shanghai-based pediatric Tuina masters,and folk techniques while incorporating his years of clinical insights.This synthesis finally led to the formulation of the academic thought of"reinforcing healthy Qi and unblocking regulation"in Shanghai-style pediatric Tuina that guides clinical practice.Specifically,he comprehensively applies techniques such as"unblocking regulation of Zang-Fu organs","unblocking regulation of the four seas","unblocking regulation of the water and fire",and"unblocking regulation of the back"to the prevention and treatment of pediatric diseases across internal medicine,external medicine,orthopedics,and otolaryngology,which has significantly enhanced clinical efficacy and expanded the applicable age range and scope of Tuina for pediatric health issues,more aligning with the characteristics of children and adolescents'health challenges and current clinical demands,and paving a new way in preserving and developing traditional pediatric Tuina.
8.Expert consensus on orthodontic treatment of protrusive facial deformities.
Jie PAN ; Yun LU ; Anqi LIU ; Xuedong WANG ; Yu WANG ; Shiqiang GONG ; Bing FANG ; Hong HE ; Yuxing BAI ; Lin WANG ; Zuolin JIN ; Weiran LI ; Lili CHEN ; Min HU ; Jinlin SONG ; Yang CAO ; Jun WANG ; Jin FANG ; Jiejun SHI ; Yuxia HOU ; Xudong WANG ; Jing MAO ; Chenchen ZHOU ; Yan LIU ; Yuehua LIU
International Journal of Oral Science 2025;17(1):5-5
Protrusive facial deformities, characterized by the forward displacement of the teeth and/or jaws beyond the normal range, affect a considerable portion of the population. The manifestations and morphological mechanisms of protrusive facial deformities are complex and diverse, requiring orthodontists to possess a high level of theoretical knowledge and practical experience in the relevant orthodontic field. To further optimize the correction of protrusive facial deformities, this consensus proposes that the morphological mechanisms and diagnosis of protrusive facial deformities should be analyzed and judged from multiple dimensions and factors to accurately formulate treatment plans. It emphasizes the use of orthodontic strategies, including jaw growth modification, tooth extraction or non-extraction for anterior teeth retraction, and maxillofacial vertical control. These strategies aim to reduce anterior teeth and lip protrusion, increase chin prominence, harmonize nasolabial and chin-lip relationships, and improve the facial profile of patients with protrusive facial deformities. For severe skeletal protrusive facial deformities, orthodontic-orthognathic combined treatment may be suggested. This consensus summarizes the theoretical knowledge and clinical experience of numerous renowned oral experts nationwide, offering reference strategies for the correction of protrusive facial deformities.
Humans
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Orthodontics, Corrective/methods*
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Consensus
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Malocclusion/therapy*
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Patient Care Planning
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Cephalometry
9.Analgesic effect of "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique of acupuncture on primary dysmenorrhea based on NOD1/RIP2/NF-κB signaling pathway in the rats.
Xu JIN ; Yanlin ZHANG ; Boya CHANG ; Jia REN ; Jianheng HAO ; Yuxia CAO ; Haijun WANG ; Laixi JI
Chinese Acupuncture & Moxibustion 2025;45(2):209-216
OBJECTIVE:
To observe the effect of "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique on the relative protein expression of the signaling pathway of nucleotide-binding oligomerization domain-containing protein 1 (NOD1)/ receptor-interacting protein 2 (RIP2)/nuclear factor kappa-B (NF-κB) and the expression of proinflammatory cytokines in the rats with primary dysmenorrhea (PD), so as to explore the underlying mechanism of this acupuncture technique for pain alleviation in PD.
METHODS:
Thirty female SD rats of SPF grade with normal estrous cycle were randomized into a blank group, a model group and an acupuncture group, 10 rats in each one. Using the intraperitoneal injection with estradiol benzoate combined with oxytocin, PD model was prepared in the model group and the acupuncture group. In the acupuncture group, during model preparation, the intervention with "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique was delivered simultaneously, 20 min each time, once daily for consecutive 10 days. On day 11, within 30 min after the intraperitoneal injection with oxytocin, the writhing reaction (latency, frequency and score) was recorded; the morphology of uterine tissue was observed with HE staining, the contents of prostaglandin E2 (PGE2), prostaglandin F2α (PGF2α), interleukin (IL)-1β, IL-18, cyclooxygenase-2 (COX-2), and tumor necrosis factor-α(TNF-α) in the serum were detected using ELISA method; the relative protein expression of NOD1, RIP2, NF-κB p65, phosphorylation-NF-κB p65 (p-NF-κB p65) was detected in the uterine tissue using Western blot method; and the mRNA expression of NOD1, RIP2 and NF-κB p65 was detected with the quantitative real-time PCR employed.
RESULTS:
Compared with the blank group, in the model group, the writhing latency was prolonged (P<0.01), the writhing frequency and score increased (P<0.01) in the rats; the endometrial epithelial cells showed massive degeneration and necrosis, with severe endometrial edema and widespread shedding, combined with neutrophil infiltration; the serum PGE2 content was dropped (P<0.01), while those of PGF2α, IL-1β, IL-18, COX-2, and TNF-α elevated (P<0.01); the protein expression of NOD1, RIP2, NF-κB p65 and p-NF-κB p65, and the mRNA expression of NOD1, RIP2 and NF-κB p65 in uterine tissue increased (P<0.01). In comparison with the model group, in the acupuncture group, the writhing latency was prolonged (P<0.01), the writhing frequency and score were reduced (P<0.01) in the rats; there was less degeneration and necrosis of endometrial epithelial cells, with mild endometrial edema and very little neutrophil infiltration; the serum PGE2 content increased (P<0.01), while those of PGF2α, IL-1β, IL-18, COX-2, and TNF-α decreased (P<0.01); the protein expression of NOD1, RIP2, NF-κB p65 and p-NF-κB p65 and the mRNA expression of NOD1, RIP2 and NF-κB p65 in uterine tissue were dropped (P<0.05, P<0.01).
CONCLUSION
"Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique can alleviate the pain symptom of PD rats, and its action mechanism may be related to inhibiting the active expression of NOD1/RIP2/NF-κB signaling pathway in the uterine tissue, thereby reducing the inflammatory response.
Animals
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Female
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Rats, Sprague-Dawley
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Rats
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Signal Transduction
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Dysmenorrhea/metabolism*
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NF-kappa B/metabolism*
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Acupuncture Points
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Humans
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Acupuncture Analgesia
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Nod1 Signaling Adaptor Protein/metabolism*
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Receptor-Interacting Protein Serine-Threonine Kinase 2/metabolism*
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Acupuncture Therapy
10.Effect of "Zhibian" (BL54) toward "Shuidao" (ST28) acupuncture on gut microbiota in mice with poor ovarian response.
Boya CHANG ; Jia REN ; Xu JIN ; Jianheng HAO ; Zhen GAO ; Yuxia CAO ; Haijun WANG
Chinese Acupuncture & Moxibustion 2025;45(6):770-780
OBJECTIVE:
To explore the possible mechanism by which the "Zhibian" (BL54) toward "Shuidao" (ST28) acupuncture improves ovarian function in mice with poor ovarian response (POR) by observing its effect on gut microbiota.
METHODS:
A total of 35 SPF-grade C57BL/6 female mice were screened for normal estrous cycles using vaginal smears, and 30 mice were selected. Ten mice were assigned to the blank group, while the remaining mice were used to establish the POR model by intragastric administration of tripterygium wilfordii suspension. The successfully modeled mice were randomly divided into a model group and an acupuncture group, with 10 mice in each group. After modeling, the acupuncture group received the "Zhibian" (BL54) toward "Shuidao" (ST28) acupuncture method once daily for 20 minutes per session. Ovulation induction began the day after the intervention, and samples were collected after ovulation induction. Vaginal cytology was used to observe estrous cycle changes, and the number of oocytes obtained, ovarian wet weight, and ovarian index were recorded. Serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), and anti-Müllerian hormone (AMH) were detected using ELISA. HE staining was used to observe ovarian histology. Gut microbiota was analyzed using 16S rRNA gene sequencing technology. Western blot was used to detect the relative protein expression levels of Occludin and zonula occludens-1 (ZO-1) in colonic tissue. Correlation analysis was conducted among serum hormone indexes, the number of oocytes obtained, ovarian index and gut microbiota.
RESULTS:
Compared with the blank group, the model group showed a higher estrous cycle disorder rate (P<0.01), increased serum FSH and LH levels, and a higher LH/FSH ratio (P<0.01), while the number of oocytes obtained, ovarian wet weight, ovarian index, and serum E2 and AMH levels were significantly reduced (P<0.01). Compared with the model group, the acupuncture group showed a lower estrous cycle disorder rate (P<0.01), decreased serum FSH and LH levels, and a lower LH/FSH ratio (P<0.01), along with an increased number of oocytes obtained, higher ovarian wet weight, ovarian index, and elevated serum AMH and E2 levels (P<0.01, P<0.05). The blank group had a large number of well-developed primordial follicles, with abundant and closely arranged follicles at various stages. In the model group, there was a significant increase in the number of atretic follicles, a reduction in the number of follicles at various stages, and loosely arranged ovarian tissue. Compared with the blank group, the model group showed a significant decrease in the number of normal follicles (P<0.01) and an increase in the number of atretic follicles (P<0.01). The acupuncture group showed a reduction in atretic follicles and an increase in the number of follicles at various stages compared with the model group, with a significant increase in normal follicles (P<0.01) and a decrease in atretic follicles (P<0.01). Compared with the blank group, the model group exhibited reduced gut microbiota diversity and richness, with significantly lower Chao1 and Shannon indices (P<0.01), and a greater clustering distance from the blank group. The model group also showed an increase in the relative abundance of Firmicutes_D, Verrucomicrobiota, Paramuribaculum, Dubosiella, and Muribaculum (P<0.01, P<0.05), while the relative abundance of Firmicutes_A and the relative protein expression of Occludin and ZO-1 in colonic tissue were decreased (P<0.01). Compared with the model group, the acupuncture group showed improved gut microbiota diversity and richness, with increased Chao1 and Shannon indices (P<0.05), and a clustering distance closer to the blank group. The acupuncture group exhibited reduced relative abundance of Firmicutes_D, Verrucomicrobiota, and Muribaculum (P<0.05, P<0.01), while the relative abundance of Firmicutes_A and the relative protein expression of Occludin and ZO-1 were significantly increased (P<0.01, P<0.05). Correlation analysis indicated a relationship between gut microbiota and serum hormone indicators, as well as the ovarian index. Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis showed that the metabolic pathways of the intersecting species were related to amino acid biosynthesis and nucleotide metabolism.
CONCLUSION
The "Zhibian" (BL54) toward "Shuidao" (ST28) acupuncture method improves ovarian function in POR mice, and its mechanism may be related to regulating gut microbiota structure and maintaining intestinal barrier homeostasis.
Animals
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Female
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Gastrointestinal Microbiome
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Mice
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Acupuncture Therapy
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Mice, Inbred C57BL
;
Humans
;
Ovary/physiopathology*
;
Acupuncture Points
;
Follicle Stimulating Hormone/metabolism*
;
Luteinizing Hormone/metabolism*
;
Estrous Cycle
;
Anti-Mullerian Hormone/blood*

Result Analysis
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