1.Mechanism of Diaphragmatic Dysfunction in Chronic Obstructive Pulmonary Disease and Treatment with Traditional Chinese Medicine: A Review
Yuanyuan YING ; Xiaoqing ZHOU ; Kaiwen NI ; Zhen WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):285-296
Chronic obstructive pulmonary disease(COPD) is a common chronic respiratory disorder frequently accompanied by diaphragmatic dysfunction during its course, which significantly increases respiratory burden and impairs quality of life. As the primary inspiratory muscle, the diaphragm is prone to fatigue, atrophy, inflammation, and fibrosis during the long-term progression of COPD. Its pathological mechanisms involve multiple pathways such as inflammatory responses, oxidative stress, mitochondrial dysfunction, apoptosis, ion channel abnormalities, epigenetic regulation, autophagy disorder, and protein metabolism imbalance. In recent years, traditional Chinese medicine(TCM) has demonstrated multi-targeted and systemic regulatory advantages in improving diaphragmatic function in COPD. However, related studies remain fragmented, and integrated mechanistic understanding is lacking. This paper focuses on the mechanism-target-TCM intervention framework, systematically summarizing the molecular mechanisms of diaphragmatic dysfunction, while incorporating the TCM theory of Zongqi(ancestral Qi). It highlights the therapeutic effects of Chinese herbal formulas, single herbs, and active components in modulating inflammation, oxidative stress, mitochondrial function, ion channels, epigenetic processes, autophagy, and protein homeostasis. Additionally, the review outlines existing challenges, including insufficient study volume, unbalanced selection of herbal prescriptions, limited mechanistic depth, inconsistent disease models and experimental designs, lack of standardized diaphragmatic function assessment, and weak clinical validation. Future research should strengthen the integration of TCM and modern medicine, identify additional therapeutic targets, deepen mechanistic research, and establish unified and standardized experimental systems to advance the theoretical foundation and clinical application of TCM in the prevention and treatment of COPD-related diaphragmatic dysfunction.
2.Mechanism of Diaphragmatic Dysfunction in Chronic Obstructive Pulmonary Disease and Treatment with Traditional Chinese Medicine: A Review
Yuanyuan YING ; Xiaoqing ZHOU ; Kaiwen NI ; Zhen WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):285-296
Chronic obstructive pulmonary disease(COPD) is a common chronic respiratory disorder frequently accompanied by diaphragmatic dysfunction during its course, which significantly increases respiratory burden and impairs quality of life. As the primary inspiratory muscle, the diaphragm is prone to fatigue, atrophy, inflammation, and fibrosis during the long-term progression of COPD. Its pathological mechanisms involve multiple pathways such as inflammatory responses, oxidative stress, mitochondrial dysfunction, apoptosis, ion channel abnormalities, epigenetic regulation, autophagy disorder, and protein metabolism imbalance. In recent years, traditional Chinese medicine(TCM) has demonstrated multi-targeted and systemic regulatory advantages in improving diaphragmatic function in COPD. However, related studies remain fragmented, and integrated mechanistic understanding is lacking. This paper focuses on the mechanism-target-TCM intervention framework, systematically summarizing the molecular mechanisms of diaphragmatic dysfunction, while incorporating the TCM theory of Zongqi(ancestral Qi). It highlights the therapeutic effects of Chinese herbal formulas, single herbs, and active components in modulating inflammation, oxidative stress, mitochondrial function, ion channels, epigenetic processes, autophagy, and protein homeostasis. Additionally, the review outlines existing challenges, including insufficient study volume, unbalanced selection of herbal prescriptions, limited mechanistic depth, inconsistent disease models and experimental designs, lack of standardized diaphragmatic function assessment, and weak clinical validation. Future research should strengthen the integration of TCM and modern medicine, identify additional therapeutic targets, deepen mechanistic research, and establish unified and standardized experimental systems to advance the theoretical foundation and clinical application of TCM in the prevention and treatment of COPD-related diaphragmatic dysfunction.
3.Pregnancy outcomes in dichorionic twins: comparing assisted reproductive technology and spontaneous conception
Jiancai YIN ; Ziyan ZHAO ; Baojing ZHAO ; Zhen YU ; Yuanyuan YANG
Acta Universitatis Medicinalis Anhui 2026;61(5):908-913
ObjectiveTo investigate the differences in pregnancy outcomes between dichorionic diamniotic (DCDA) twin pregnancies conceived via assisted reproductive technology (ART) and those from spontaneous conception, in order to provide evidence for the perinatal management of ART-conceived DCDA twins. MethodsA retrospective analysis was conducted on the clinical data of 549 DCDA twin parturients. Based on the mode of conception, they were divided into ART group (423 cases) and spontaneous conception group (126 cases). The baseline characteristics, pregnancy complications and neonatal outcomes were compared between the two groups. ResultsRegarding baseline characteristics, the ART group had higher maternal age, number of previous abortions, proportions of primiparas, proportions of advanced maternal age and greater 24-hour postpartum blood loss, but lower parity and proportion of scarred uterus compared to the spontaneous conception group (P<0.05). For pregnancy complications, the ART group exhibited higher incidences of hypertensive disorders of pregnancy, placental adhesion, postpartum hemorrhage and a higher application rate of uterine artery ascending branch ligation (P<0.05). There were no statistically significant differences in neonatal-related indicators between the two groups. After adjusting for confounding factors such as age, number of abortions and parities, ART remained an independent risk factor for hypertensive disorders of pregnancy, placental adhesion and postpartum hemorrhage (P<0.05), while it did not increase the risk of adverse neonatal outcomes. Further subgroup analysis by gestational age showed that in the late preterm subgroup, the risk of ART-related pregnancy complications remained elevated (P<0.05). Additionally, neonatal outcomes showed no significant difference between the two groups in both the late preterm and term subgroups. ConclusionART increases the risk of hypertensive disorders of pregnancy, placental adhesion and postpartum hemorrhage in DCDA twin parturients. Clinically, enhanced monitoring of blood pressure during pregnancy and blood loss during delivery and postpartum, along with targeted prevention and management of these complications, are crucial for improving pregnancy outcomes.
4.Comparative Analysis of Sex Differences Between Cyclophosphamide- and Paclitaxel-Induced Mouse Models of Chemotherapy-Induced Alopecia
Qiuming ZHONG ; Liping QIAN ; Yuanyuan CAI ; Deqiang LIU ; Wei LIN ; Chunying YU ; Jindong XIE ; Xunli WANG ; Sheng WU ; Biyin CHEN ; Zhen HUANG
Laboratory Animal and Comparative Medicine 2026;46(5):677-688
ObjectiveTo evaluate the effect of sex on phenotypic stability in mouse models of chemotherapy-induced alopecia (CIA) induced by cyclophosphamide (CYP) or paclitaxel (PTX), and to explore potential mechanisms underlying phenotypic heterogeneity in female mice. MethodsTwenty-eight wild-type specific pathogen-free (SPF) C57BL/6J mice (half male and half female) aged 6-8 weeks were selected for synchronous back hair removal. On the 9th day after hair removal (T0), the mice were randomly divided into a wild-type female CYP group, a wild-type male CYP group, a female negative control group and a male negative control group. Mice in the CYP groups were intraperitoneally injected with a single dose of 120 mg/kg CYP, and mice in the negative control groups were intraperitoneally injected with the same volume of normal saline. Subsequently, the phenotypic changes of the CIA model induced by CYP were continuously observed and photographed. Eleven 6-week-old SPF female C57BL/6J mice underwent bilateral ovariectomy (OVX group). Following successful surgery, these mice and five XY (Sry⁻) mice bred from Four Core Genotypes breeding stock were used to establish the chemotherapy-induced alopecia (CIA) model using the same protocol. Serum was collected from mice in the OVX group, wild-type female CYP group, and wild-type male CYP group on T7 after CYP administration for steroid hormone analysis by mass spectrometry to evaluate the effects of gonadal estrogen and sex chromosome background changes on the stability of the CYP-induced female alopecic phenotype. Finally, twenty SPF C57BL/6J mice aged 6-8 weeks (half male and half female) were randomly divided into a PTX low-dose group and a PTX high-dose group. On the 9th day after simultaneous hair removal, PTX was injected intraperitoneally at a single dose of 80 mg/kg or 120 mg/kg, respectively. Mice from the PTX low-dose group and the PTX high-dose group were continuously observed and photographed to compare the intra-group consistency of the phenotypes of female and male mice. Six SPF C57BL/6J mice (half male and half female) aged 6-8 weeks were used to repeat the construction method of the PTX low-dose group, and the hair follicle histological examination was performed at T0, T4, T7, T14 and T21, respectively, to observe the damage caused by PTX to hair follicle regeneration. ResultsThe phenotypes of CYP-induced male CIA mice are stable and reproducible. There are obvious individual differences in the alopecia phenotype in the CYP-induced female CIA model mice, which may be mainly caused by the differences in the relevant estrogens in the estrous cycle. The difference between OVX+CYP group and XY (Sry⁻) +CYP group disappeared, suggesting that ovarian-derived hormones and XY (Sry⁻) sex chromosome background may be involved in the formation of phenotypic heterogeneity of CYP-induced alopecia in female mice. The mice in the high-dose PTX group and the low-dose PTX group showed a relatively consistent alopecia and regrowth process, and there was no significant sex difference. ConclusionCYP-induced CIA is strongly influenced by sex, and ovariectomy or alteration of the sex chromosome background can improve the consistency of CYP-induced alopecic phenotypes in female mice. PTX-induced CIA shows good phenotypic consistency in both female and male mice, indicating broad applicability across sexes. Taken together, the choice of animal sex in CIA model construction, mechanistic studies, and evaluation of anti-alopecia interventions should take into account the type of chemotherapeutic agent, the study objective, and the required degree of phenotypic stability. Sex should therefore be considered an important variable in model standardization and interpretation of experimental findings.
5.Research progress in antibody drug therapy for relapsed/refractory diffuse large B-cell lymphoma
Yanyan SUN ; Weichen ZHAO ; Chunyuan HE ; Yimiao XIA ; Wei ZHOU ; Yuanyuan ZHEN ; Junjie JIANG ; Facai WANG
China Pharmacy 2025;36(13):1677-1682
Diffuse large B-cell lymphoma(DLBCL)is a highly heterogeneous disease.Although standard first-line regimens can cure>50%of patients,approximately one-third of them develop relapsed/refractory DLBCL(r/r DLBCL).Consequently,immunotherapy targeting molecular abnormalities has become pivotal for managing r/r DLBCL.The results of this review show that with advances in understanding DLBCL pathogenesis and the tumor immune microenvironment,antibody-based therapies have evolved rapidly,progressing from monoclonal antibodies(e.g.,rituximab,tafasitamab)to bispecific antibodies(e.g.,odronextamab,glofitamab,epcoritamab)and antibody-drug conjugate(e.g.,polatuzumab vedotin,loncastuximab tesirine).These engineered agents enhance immune cytotoxicity and tumor-specific targeting,providing novel therapeutic options for r/r DLBCL patients.
6.Transcutaneous auricular vagus nerve stimulation regulates functional connectivity of thalamic subregions in patients with premenstrual syndrome
Ruijing SUN ; Yinqi LAI ; Ya CHEN ; Yuejuan WU ; Zhen LIU ; Qingping ZHANG ; Ziyan LAI ; Gaoxiong DUAN ; Yan ZHANG ; Shanshan LI ; Yuanyuan OU ; Sijing TUO ; Hui ZHOU ; Rongcai WU ; Zhizhong CHEN ; Demao DENG
Chinese Journal of Radiology 2025;59(12):1384-1392
Objective:To investigate the regulatory effects of transcutaneous auricular vagus nerve stimulation (taVNS) on functional connectivity (FC) of thalamic subregions in patients with premenstrual syndrome (PMS).Methods:This study was a cross-sectional investigation. Clinical, laboratory, and imaging data were retrospectively collected from 56 PMS patients (PMS group) and 66 healthy controls (control group) recruited from various universities and hospitals in Nanning between November 2021 and June 2024. Resting-state functional MRI (fMRI) data and fMRI data during taVNS immediate stimulation (2 Hz, 25 Hz) were acquired from subjects during their late luteal phase. Using thalamic subregions (anterior thalamic nucleus, lateral nucleus, ventral nucleus, medial nucleus, central nucleus, posterior nucleus) as seeds, two-sample t-tests or paired t-tests were employed to analyze alterations in thalamic subregion FC in PMS patients and the regulatory effects of taVNS on these changes. Independent samples t-test were used to compare the differences in clinical and laboratory indicators between the PMS group and the control group. The relationship between taVNS regulation of thalamic subregion FC in PMS patients and thalamic internal functional connectivity were analyzed using mediation effect analysis. Results:Compared to the control group, patients in the PMS group showed increased scores on the Daily Record of Severity of Problems, Pittsburgh Sleep Quality Index, Self-Rating Anxiety Scale, Self-Rating Depression Scale, Hamilton Anxiety Rating Scale 17, and Hamilton Depression Rating Scale 14 during the late luteal phase ( P<0.05). At baseline, PMS patients exhibited higher FC between the left thalamic lateral nucleus and the left insula, and lower FC between the left medial nucleus, posterior nucleus, and ventral nucleus of the thalamus and the right middle frontal gyrus (MFG) compared to the control group (GRF corrected, voxel-level P<0.001, cluster-level P<0.05). During 2 Hz taVNS immediate stimulation in PMS group, FC between the left thalamic medial nucleus, posterior nucleus, ventral nucleus and the right MFG, as well as the FC between the left thalamic ventral nucleu and the left MFG increased compared to baseline levels; meanwhile, FC between the left thalamic posterior nucleus, ventral nucleus and the left insula decreased compared to baseline levels (GRF corrected, voxel-level P<0.001, cluster-level P<0.05). During 25 Hz taVNS immediate stimulation, the FC between the left thalamic ventral nucleus and the right MFG decreased compared to the baseline level (GRF corrected, voxel-level P<0.001, cluster-level P<0.05). Mediation effect analysis showed that the FC between the left thalamic posterior nucleus and the left lateral nucleus mediated part of the association between the FC of the left lateral thalamic nucleus-left insula and the FC of the left ventral thalamic nucleus-left putamen/insula; there were significant direct effects between the FC of the left lateral thalamic nucleus-the left posterior nucleus and FC of the left lateral thalamic nucleus-the left insula, as well as between the FC of the left ventral thalamic nucleus-the left MFG and FC of the left ventral thalamic nucleus-the right MFG. Conclusions:taVNS can modulate abnormal FC of the left thalamic subregions in PMS patients, restoring it toward normalization. The regulatory effects of 2 Hz stimulation are more pronounced than those of 25 Hz stimulation. This modulation primarily operates through two pathways: the left thalamic lateral nucleus-left insula-left thalamic ventral nucleus pathway and the left MFG-left thalamic ventral nucleus-right MFG.
7.Impact of "Internet +" empowerment education based on timing it right on psychological craving, anxiety symptoms and relapse rates in patients with alcohol dependence
Hao WANG ; Wei LI ; Wen'ge ZHEN ; Yuanyuan LI ; Jie LIU
Sichuan Mental Health 2025;38(1):34-40
BackgroundAlcohol dependence patients are prone to relapse after their attempts to quit drinking, which poses a considerable threat to their physical and mental health and creates a heavy burden on their families. Currently, empowerment education is increasingly being utilized in the rehabilitation management of chronic diseases, but there remains a striking lack of empirical research on the application of "Internet +" empowerment education based on timing it right in alcohol dependence patients. ObjectiveTo explore the impact of "Internet +" empowerment education based on timing it right on patients with alcohol dependence, in order to maximize the reduction in relapse rates, craving for alcohol and severity of anxiety symptoms. MethodsA total of 120 patients who were hospitalized in the Department of Addiction Medicine, Hebei Provincial Mental Health Center from May 2022 to April 2023 and met the diagnostic criteria for alcohol dependence in the International Classification of Diseases, tenth edition (ICD-10) were enrolled, and they were classified into study group (n=62) and control group (n=58) using random number table methods. Both groups received standard medication and routine care. Additionally, study group underwent a 6-month "Internet +" empowerment education based on timing it right. At baseline, all subjects were assessed using Penn Alcohol Craving Scale (PACS) and Self-rating Anxiety Scale (SAS). Three months and six months after intervention, assessments were conducted using PACS, SAS and Michigan Alcoholism Screening Test (MAST). ResultsThe relapse rates after three and six months of intervention were both lower in study group than those in control group, with statistically significant differences (χ2=8.575, 8.828, P<0.01). ANOVA with repeated measures on PACS total score and scores of each item revealed a significant time effect, group effect and time×group interaction effect (F=159.714~837.751, 84.645~393.606, 24.302~137.896, P<0.01). And significant time effect, group effect and time×group interaction effect were also reported on SAS scores (F=166.237, 65.325, 24.724, P<0.01). Conclusion"Internet +" empowerment education based on timing it right may help reduce relapse rates, alcohol cravings and severity of anxiety symptoms among patients with alcohol dependence. [Funded by 2023 Annual Hebei Provincial Medical Scientific Research Project Plan (number, 20231537)]
8.Transcutaneous auricular vagus nerve stimulation regulates functional connectivity of thalamic subregions in patients with premenstrual syndrome
Ruijing SUN ; Yinqi LAI ; Ya CHEN ; Yuejuan WU ; Zhen LIU ; Qingping ZHANG ; Ziyan LAI ; Gaoxiong DUAN ; Yan ZHANG ; Shanshan LI ; Yuanyuan OU ; Sijing TUO ; Hui ZHOU ; Rongcai WU ; Zhizhong CHEN ; Demao DENG
Chinese Journal of Radiology 2025;59(12):1384-1392
Objective:To investigate the regulatory effects of transcutaneous auricular vagus nerve stimulation (taVNS) on functional connectivity (FC) of thalamic subregions in patients with premenstrual syndrome (PMS).Methods:This study was a cross-sectional investigation. Clinical, laboratory, and imaging data were retrospectively collected from 56 PMS patients (PMS group) and 66 healthy controls (control group) recruited from various universities and hospitals in Nanning between November 2021 and June 2024. Resting-state functional MRI (fMRI) data and fMRI data during taVNS immediate stimulation (2 Hz, 25 Hz) were acquired from subjects during their late luteal phase. Using thalamic subregions (anterior thalamic nucleus, lateral nucleus, ventral nucleus, medial nucleus, central nucleus, posterior nucleus) as seeds, two-sample t-tests or paired t-tests were employed to analyze alterations in thalamic subregion FC in PMS patients and the regulatory effects of taVNS on these changes. Independent samples t-test were used to compare the differences in clinical and laboratory indicators between the PMS group and the control group. The relationship between taVNS regulation of thalamic subregion FC in PMS patients and thalamic internal functional connectivity were analyzed using mediation effect analysis. Results:Compared to the control group, patients in the PMS group showed increased scores on the Daily Record of Severity of Problems, Pittsburgh Sleep Quality Index, Self-Rating Anxiety Scale, Self-Rating Depression Scale, Hamilton Anxiety Rating Scale 17, and Hamilton Depression Rating Scale 14 during the late luteal phase ( P<0.05). At baseline, PMS patients exhibited higher FC between the left thalamic lateral nucleus and the left insula, and lower FC between the left medial nucleus, posterior nucleus, and ventral nucleus of the thalamus and the right middle frontal gyrus (MFG) compared to the control group (GRF corrected, voxel-level P<0.001, cluster-level P<0.05). During 2 Hz taVNS immediate stimulation in PMS group, FC between the left thalamic medial nucleus, posterior nucleus, ventral nucleus and the right MFG, as well as the FC between the left thalamic ventral nucleu and the left MFG increased compared to baseline levels; meanwhile, FC between the left thalamic posterior nucleus, ventral nucleus and the left insula decreased compared to baseline levels (GRF corrected, voxel-level P<0.001, cluster-level P<0.05). During 25 Hz taVNS immediate stimulation, the FC between the left thalamic ventral nucleus and the right MFG decreased compared to the baseline level (GRF corrected, voxel-level P<0.001, cluster-level P<0.05). Mediation effect analysis showed that the FC between the left thalamic posterior nucleus and the left lateral nucleus mediated part of the association between the FC of the left lateral thalamic nucleus-left insula and the FC of the left ventral thalamic nucleus-left putamen/insula; there were significant direct effects between the FC of the left lateral thalamic nucleus-the left posterior nucleus and FC of the left lateral thalamic nucleus-the left insula, as well as between the FC of the left ventral thalamic nucleus-the left MFG and FC of the left ventral thalamic nucleus-the right MFG. Conclusions:taVNS can modulate abnormal FC of the left thalamic subregions in PMS patients, restoring it toward normalization. The regulatory effects of 2 Hz stimulation are more pronounced than those of 25 Hz stimulation. This modulation primarily operates through two pathways: the left thalamic lateral nucleus-left insula-left thalamic ventral nucleus pathway and the left MFG-left thalamic ventral nucleus-right MFG.
9.Value of serum 25-hydroxyvitamin D,heart-type fatty acid-binding protein and N-terminal pro-brain natriuretic peptide in assessing early myocardial injury in patients with acute exacerbation of chronic obstructive pulmonary disease
Dongge CHANG ; Zhen SUN ; Shaohua ZHANG ; Xiaofeng LIU ; Yuanyuan SU
Journal of Clinical Medicine in Practice 2025;29(14):94-98,103
Objective To investigate the evaluation value of serum 25-hydroxyvitamin D[25-(OH)D],heart-type fatty acid-binding protein(H-FABP),and N-terminal pro-brain natriuret-ic peptide(NT-ProBNP)in early myocardial injury in patients with acute exacerbation of chronic ob-structive pulmonary disease(AECOPD).Methods A total of 120 patients with AECOPD(AECOPD group)were enrolled in this study.Based on the presence of early myocardial injury,they were divided into injury group(n=68)and non-injury group(n=52).Additionally,40 healthy individuals undergoing physical examinations during the same period were included as control group.The differences in serum 25-(OH)D,H-FABP,and NT-ProBNP levels were compared,and the correlations between these markers and clinical data were analyzed.Binary logistic regression analysis was used to explore the relationships between these markers and the occurrence of early myocardial injury.Receiver op-erating characteristic(ROC)curve analysis was employed to assess the diagnostic value of these markers for early myocardial injury in AECOPD patients.Results The forced expiratory volume in the first second(FEV1),forced vital capacity(FVC),the ratio of FEV1 to FVC(FEV1/FVC),and arterial partial pressure of oxygen[pa(O2)]levels in the AECOPD group were lower than those in the control group,while the arterial partial pressure of carbon dioxide[p a(CO2)]level was high-er,with statistically significant differences(P<0.05).The serum 25-(OH)D levels in the AECOPD group and the injury group were lower than those in the control group and the non-injury group,re-spectively,while the H-FABP and NT-ProBNP levels were higher,with statistically significant differences(P<0.05).In AECOPD patients,serum 25-(OH)D was positively correlated with FEV1,FVC,FEV1/FVC,and pa(O2),and negatively correlated with pa(CO2)(P<0.05).In contrast,H-FABP and NT-ProBNP were negatively correlated with FEV1,FVC,FEV1/FVC,and pa(O2),and positively correlated with pa(CO2)(P<0.05).Binary Logistic regression analysis revealed that 25-(OH)D,H-FABP,and NT-ProBNP were related influencing factors for early myo-cardial injury in AECOPD patients(P<0.05).ROC curve analysis showed that the areas under the curve(AUCs)for evaluating myocardial injury status based on 25-(OH)D,H-FABP,and NT-ProBNP values were 0.814,0.959,and 0.837,respectively.The AUC of their combination was 0.983,with a sensitivity of 97.06%and a specificity of 80.77%.Conclusion During early myocardial injury in AECOPD patients,there is low expression of serum 25-(OH)D and high ex-pression of H-FABP and NT-ProBNP.These three markers are correlated with early myocardial inju-ry and can serve as reference indicators for clinical diagnosis.
10.Efficacy and Mechanism of Shuanghua Drink in Treating Primary Dysmenorrhea Based on COX-2/NF-κB Signaling Pathway
Yuncheng MA ; Yuanyuan SHI ; Zhen LIU ; Yuxi WANG ; Yuan TIAN ; Qian LI ; Xiaozhu WANG ; Cheng HE ; Wenhui XU ; Weiling WANG ; Jian GAO ; Ting WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):72-80
ObjectiveTo evaluate the efficacy of Shuanghua drink in treating primary dysmenorrhea in the rat model and explore its mechanism of action. MethodsAn oxytocin-induced writhing mouse model was established to evaluate the analgesic effect of Shuanghua drink. Forty-eight non-pregnant female institute of cancer research (ICR) mice were randomly divided into six groups, including a blank group, a model group, an ibuprofen group (85.00 mg·kg-1), a low-dose group of Shuanghua drink (7.14 mL·kg-1), a medium-dose group of Shuanghua drink (14.28 mL·kg-1), and a high-dose group of Shuanghua drink (28.57 mL·kg-1). Each group consisted of eight mice. All treatment groups received daily intragastric administration at corresponding doses for 10 consecutive days. One hour after the final administration, 2 U of oxytocin was intraperitoneally injected per mouse. The writhing latency and number of writhing within 20 minutes were recorded. A primary dysmenorrhea rat model was established by using estradiol benzoate and oxytocin to evaluate the inhibitory effect of Shuanghua drink on the contraction of uterine smooth muscle. Forty-eight non-pregnant female Sprague-Dawley (SD) rats were divided into six groups, including a blank group, a model group, an ibuprofen group (51.00 mg·kg-1), a low-dose group of Shuanghua drink (4.28 mL·kg-1), a medium-dose group of Shuanghua drink (8.57 mL·kg-1), and a high-dose group of Shuanghua drink (17.10 mL·kg-1). Each group consisted of eight rats. Rats received subcutaneous injections of estradiol benzoate for 10 consecutive days to enhance uterine sensitivity. On the eleventh day, oxytocin (2 U/rat) was intraperitoneally administered to induce abnormal uterine contractions for establishing the primary dysmenorrhea model. All treatment groups received daily intragastric administration from the second day of modeling for 10 days. The effects of Shuanghua drink were evaluated by using parameters including uterine motility and the variation rate of uterine motility. The mechanism of action was investigated in rats with primary dysmenorrhea. The content of prostaglandin F2α (PGF2α), prostaglandin E2 (PGE2), thromboxane B2 (TXB2), prostacyclin metabolite (6-keto-PGF1α), and β-endorphin (β-EP) in uterine tissue of rats was detected by using enzyme-linked immunosorbent assay (ELISA). The changes in the content of nitric oxide (NO) and inducible nitric oxide synthase (iNOS) were analyzed via colorimetric assay. Western blot was performed to determine the content of phosphorylated inhibitor of kappa B kinase beta (p-IKKβ)/IKKβ, phosphorylated inhibitor of kappa B alpha (p-IκBα), IκBα, phosphorylated p65 (p-p65), p65, and cyclooxygenase-2 (COX-2) proteins in uterine tissue of rats. ResultsIn the oxytocin-induced writhing mouse model, the model group exhibited significantly shortened writhing latency and increased writhing frequency compared to the control group (P<0.01). Both the ibuprofen group and the high-dose group of Shuanghua drink displayed prolonged writhing latency (P<0.05), while the ibuprofen group and the low-dose, medium-dose, and high-dose groups of Shuanghua drink exhibited reduced writhing frequency (P<0.01). In the primary dysmenorrhea rat model, the uterine motility and its variation rate in the model group were significantly higher than those in the blank group (P<0.01). These parameters were markedly suppressed by ibuprofen and Shuanghua drink at all tested doses (P<0.01). For the mechanism of action, the model group showed significantly increased PGF2α/PGE2, TXB2/6-keto-PGF1α, NO, and iNOS in uterine tissue (P<0.05, P<0.01) and significantly decreased β-EP (P<0.01). These parameters were significantly attenuated in the ibuprofen group and the low-dose, medium-dose, and high-dose groups of Shuanghua drink. The PGF2α/PGE2 (P<0.01), TXB2/6-keto-PGF1α (P<0.01), NO (medium-dose group P<0.05), and iNOS (P<0.01) were reduced, and the β-EP (medium-dose group P<0.05) was up-regulated. Compared to the model group, the ibuprofen group and medium-dose group of Shuanghua drink showed significantly increased content of β-EP in the serum of rats (P<0.05). Compared to the blank group, the model group showed significantly elevated expressions of COX-2, p-IKKβ/IKKβ, p-IκBα/IκBα, and p-p65/p65 proteins (P<0.01) and significantly reduced anti-inflammatory protein IκBα (P<0.05). Compared to the model group, the ibuprofen group and the low-dose, medium-dose, and high-dose groups of Shuanghua drink showed significantly reduced expressions of COX-2 (P<0.01), p-IKKβ/IKKβ (P<0.01), p-IκBα/IκBα (P<0.05, P<0.01), and p-p65/p65(P<0.01) and up-regulated expression of IκBα protein (P<0.05, P<0.01). ConclusionShuanghua drink effectively alleviates primary dysmenorrhea through analgesia and suppression of abnormal contractions of uterine smooth muscle. Its mechanism may be mediated by reduced levels of PGF2α/PGE2, TXB2/6-keto-PGF1α, iNOS, and NO, elevated β-EP level, and inhibited COX-2/NF-κB signaling pathway.

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