1.Effects of human umbilical cord blood platelet-rich plasma,mononuclear cells,and mesenchymal stem cells in repairing thin endometrium in rats
Yanli MU ; Anchun HU ; Wenchi XU ; Panpan CHEN ; Hao CHEN ; Shuyun ZHAO ; Guanyou HUANG ; Xiaojuan CHEN
Chinese Journal of Tissue Engineering Research 2026;30(1):78-92
BACKGROUND:Research has found that human umbilical cord blood platelet rich plasma and human umbilical cord blood derived mesenchymal stem cells have certain therapeutic effects on thin endometrium.However,there are currently no reports on the study of human umbilical cord blood mononuclear cells on thin endometrium,and there is a lack of relevant research comparing the three.OBJECTIVE:To explore the effects and mechanisms of human umbilical cord blood platelet rich plasma,monocytes,and mesenchymal stem cells in repairing thin endometrium in rats.METHODS:Sixty female SPF grade SD rats were randomly divided into sham operation group,model group,human umbilical cord blood platelet rich plasma group,human umbilical cord blood mononuclear cell group,and human umbilical cord blood derived mesenchymal stem cell group,with 12 rats in each group.The sham operation group received 0.5 mL physiological saline injection into the uterine horn,followed by 0.5 mL of PBS infusion after 5 minutes;The model group,human umbilical cord blood platelet rich plasma group,human umbilical cord blood mononuclear cell group,and human umbilical cord blood derived mesenchymal stem cell group were injected with 0.5 mL of 95%ethanol by volume.After 5 minutes,the remaining ethanol was aspirated and washed twice with physiological saline.Then,0.5 mL of PBS,human umbilical cord blood platelet rich plasma,human umbilical cord blood mononuclear cells(1×107 cells/mL),and human umbilical cord blood derived mesenchymal stem cells(1×107 cells/mL)were perfused separately.During the third normal estrus cycle after reperfusion,organs,tissues and serum were collected for testing of relevant indicators.RESULTS AND CONCLUSION:(1)The macroscopic view of uterine tissue,hematoxylin eosin staining and Masson staining results:the sham operation group had intact structure,moderate endometrial thickness,and clear vascular structure.Compared with the sham operation group,the model group showed uterine atrophy,incomplete structure,significantly reduced endometrial thickness and glandular quantity,disordered vascular structure,and increased fibrosis.Compared with the model group,after treatment with human umbilical cord blood derivatives,the size,structure,and endometrial thickness of the uterus were restored(all P<0.01),and fibrosis was reduced,with the most significant recovery observed in the human umbilical cord blood mononuclear cell group.The increase in glandular quantity was most significant in the human umbilical cord blood platelet rich plasma group(P<0.000 1).(2)The immunohistochemistry and immunofluorescence results of uterine tissue showed that compared with the sham operation group,the expression levels of cell proliferation related indicators such as keratin 9 and vimentin,endometrial receptivity related indicators such as leukemia inhibitory factor and integrin αyβ3,platelet endothelial cell adhesion molecule,basic fibroblast growth factor,and vascular endothelial growth factor were all reduced in the model group(all P<0.05).Compared with the model group,the above indicators were significantly increased after treatment with human umbilical cord blood derivatives.Comparison of human umbilical cord blood derivatives among groups showed that keratin 9 and vascular endothelial growth factor protein:human umbilical cord blood mononuclear cell group>human umbilical cord blood derived mesenchymal stem cell group>human umbilical cord blood platelet rich plasma group;Wave shaped protein and leukemia inhibitory factor protein:human umbilical cord blood derived mesenchymal stem cell group>human umbilical cord blood mononuclear cell group>human umbilical cord blood platelet rich plasma group;Integrin αyβ3 protein:human umbilical cord blood platelet rich plasma group>human umbilical cord blood derived mesenchymal stem cell group>human umbilical cord blood mononuclear cell group;Platelet endothelial cell adhesion molecule protein:human umbilical cord blood platelet rich plasma group>human umbilical cord blood mononuclear cell group>human umbilical cord blood derived mesenchymal stem cell group;Basic fibroblast growth factor protein:human umbilical cord blood mononuclear cell group>human umbilical cord blood platelet rich plasma group>human umbilical cord blood derived mesenchymal stem cell group.(3)Western blot analysis showed that compared with the sham operation group,the protein levels of interleukin-6,interleukin-1β,and tumor necrosis factor alpha in the model group were significantly increased(all P<0.001),and their expression levels decreased after treatment(all P<0.05).(4)ELISA assay showed that compared with the sham operation group,the model group had lower levels of anti Mullerian hormone,estradiol,and progesterone,and increased levels of follicle stimulating hormone and luteinizing hormone(except for luteinizing hormone,all P<0.05).After treatment,there was a certain degree of recovery in the levels of sex hormones and anti Mullerian hormones.(5)Fertility experiments showed that compared with the sham operation group,the model group had an increase in conception time and a significant decrease in litter size(all P<0.05).After treatment with human umbilical cord blood derivatives,the litter size of all three groups increased(P<0.05),and no significant differences were found between the groups.This study preliminarily reveals that human umbilical cord blood mononuclear cells have a certain therapeutic effect on thin endometrium,and human umbilical cord blood platelet rich plasma,human umbilical cord blood mononuclear cells,and human umbilical cord blood derived mesenchymal stem cells have different advantages and differences in improving endometrial regeneration function,endometrial receptivity,angiogenesis,inflammation regulation,and improving pregnancy outcomes in thin endometrium.
2.Effects of human umbilical cord blood platelet-rich plasma,mononuclear cells,and mesenchymal stem cells in repairing thin endometrium in rats
Yanli MU ; Anchun HU ; Wenchi XU ; Panpan CHEN ; Hao CHEN ; Shuyun ZHAO ; Guanyou HUANG ; Xiaojuan CHEN
Chinese Journal of Tissue Engineering Research 2026;30(1):78-92
BACKGROUND:Research has found that human umbilical cord blood platelet rich plasma and human umbilical cord blood derived mesenchymal stem cells have certain therapeutic effects on thin endometrium.However,there are currently no reports on the study of human umbilical cord blood mononuclear cells on thin endometrium,and there is a lack of relevant research comparing the three.OBJECTIVE:To explore the effects and mechanisms of human umbilical cord blood platelet rich plasma,monocytes,and mesenchymal stem cells in repairing thin endometrium in rats.METHODS:Sixty female SPF grade SD rats were randomly divided into sham operation group,model group,human umbilical cord blood platelet rich plasma group,human umbilical cord blood mononuclear cell group,and human umbilical cord blood derived mesenchymal stem cell group,with 12 rats in each group.The sham operation group received 0.5 mL physiological saline injection into the uterine horn,followed by 0.5 mL of PBS infusion after 5 minutes;The model group,human umbilical cord blood platelet rich plasma group,human umbilical cord blood mononuclear cell group,and human umbilical cord blood derived mesenchymal stem cell group were injected with 0.5 mL of 95%ethanol by volume.After 5 minutes,the remaining ethanol was aspirated and washed twice with physiological saline.Then,0.5 mL of PBS,human umbilical cord blood platelet rich plasma,human umbilical cord blood mononuclear cells(1×107 cells/mL),and human umbilical cord blood derived mesenchymal stem cells(1×107 cells/mL)were perfused separately.During the third normal estrus cycle after reperfusion,organs,tissues and serum were collected for testing of relevant indicators.RESULTS AND CONCLUSION:(1)The macroscopic view of uterine tissue,hematoxylin eosin staining and Masson staining results:the sham operation group had intact structure,moderate endometrial thickness,and clear vascular structure.Compared with the sham operation group,the model group showed uterine atrophy,incomplete structure,significantly reduced endometrial thickness and glandular quantity,disordered vascular structure,and increased fibrosis.Compared with the model group,after treatment with human umbilical cord blood derivatives,the size,structure,and endometrial thickness of the uterus were restored(all P<0.01),and fibrosis was reduced,with the most significant recovery observed in the human umbilical cord blood mononuclear cell group.The increase in glandular quantity was most significant in the human umbilical cord blood platelet rich plasma group(P<0.000 1).(2)The immunohistochemistry and immunofluorescence results of uterine tissue showed that compared with the sham operation group,the expression levels of cell proliferation related indicators such as keratin 9 and vimentin,endometrial receptivity related indicators such as leukemia inhibitory factor and integrin αyβ3,platelet endothelial cell adhesion molecule,basic fibroblast growth factor,and vascular endothelial growth factor were all reduced in the model group(all P<0.05).Compared with the model group,the above indicators were significantly increased after treatment with human umbilical cord blood derivatives.Comparison of human umbilical cord blood derivatives among groups showed that keratin 9 and vascular endothelial growth factor protein:human umbilical cord blood mononuclear cell group>human umbilical cord blood derived mesenchymal stem cell group>human umbilical cord blood platelet rich plasma group;Wave shaped protein and leukemia inhibitory factor protein:human umbilical cord blood derived mesenchymal stem cell group>human umbilical cord blood mononuclear cell group>human umbilical cord blood platelet rich plasma group;Integrin αyβ3 protein:human umbilical cord blood platelet rich plasma group>human umbilical cord blood derived mesenchymal stem cell group>human umbilical cord blood mononuclear cell group;Platelet endothelial cell adhesion molecule protein:human umbilical cord blood platelet rich plasma group>human umbilical cord blood mononuclear cell group>human umbilical cord blood derived mesenchymal stem cell group;Basic fibroblast growth factor protein:human umbilical cord blood mononuclear cell group>human umbilical cord blood platelet rich plasma group>human umbilical cord blood derived mesenchymal stem cell group.(3)Western blot analysis showed that compared with the sham operation group,the protein levels of interleukin-6,interleukin-1β,and tumor necrosis factor alpha in the model group were significantly increased(all P<0.001),and their expression levels decreased after treatment(all P<0.05).(4)ELISA assay showed that compared with the sham operation group,the model group had lower levels of anti Mullerian hormone,estradiol,and progesterone,and increased levels of follicle stimulating hormone and luteinizing hormone(except for luteinizing hormone,all P<0.05).After treatment,there was a certain degree of recovery in the levels of sex hormones and anti Mullerian hormones.(5)Fertility experiments showed that compared with the sham operation group,the model group had an increase in conception time and a significant decrease in litter size(all P<0.05).After treatment with human umbilical cord blood derivatives,the litter size of all three groups increased(P<0.05),and no significant differences were found between the groups.This study preliminarily reveals that human umbilical cord blood mononuclear cells have a certain therapeutic effect on thin endometrium,and human umbilical cord blood platelet rich plasma,human umbilical cord blood mononuclear cells,and human umbilical cord blood derived mesenchymal stem cells have different advantages and differences in improving endometrial regeneration function,endometrial receptivity,angiogenesis,inflammation regulation,and improving pregnancy outcomes in thin endometrium.
3.Comparative Study on the Efficacy of Traditional Mongolian Medicine Bone Setting and Surgical Treatment for Humeral Surgical Neck Fractures
Jin Ai Hua ; ; Ba Hu Shan ; Ta Na ; Wu Da Mu ; Bao Tu Ya ; Si Qin ; Tsend-Ayush D ; Bolortulga Z
Mongolian Journal of Health Sciences 2026;91(1):38-42
Background:
The surgical neck of the humerus is located approximately 2–3 cm distal to the anatomical neck (collum anatomicum) at the junction of the greater and lesser tubercles and the humeral shaft. Owing to the transition between cortical and cancellous bones, this region is biomechanically vulnerable and prone to fractures, particularly in older adults. Fractures of the surgical neck of the humerus are classified under the International Classification of Diseases, 10th Revision (ICD-10), with diagnostic codes S42.21.3, S42.22, and S42.23. National statistics indicate that 210,763 new trauma cases were reported in Mongolia in 2023, representing an 11.6% increase compared to the previous year, of which 11,452 cases (5.4%) involved shoulder and humeral injuries.
Aim:
This study aimed to compare the clinical effects of traditional Mongolian bone-setting therapy and surgical treatment on pain intensity, shoulder function, range of motion, and activities of daily living in patients with surgical neck of the humerus fractures.
Materials and Methods:
Based on the Department of Traditional Therapies of the Mongolian National University of Medical Sciences (MNUMS) and the Department of Orthopedic Trauma of the Inner Mongolia International Hospital, Inner Mongolia Autonomous Region, China, a total of 60 patients diagnosed with surgical neck fractures of the humerus were enrolled in this study between January 2024 and December 2025. The patients were randomly assigned into two groups: a Mongolian traditional bone-setting therapy group (n=30) and a surgical treatment group (n=30). Assessments were conducted before treatment and at 1 month and 3 months after treatment. Shoulder joint range of motion, activities of daily living, and muscle strength were evaluated using the Constant–Murley score, while pain intensity was assessed using the Visual Analog Scale (VAS). The collected data were systematically analyzed, and the therapeutic outcomes of the two treatment modalities were comprehensively compared and summarized.
Result:
The baseline demographic and clinical characteristics were comparable between the groups. In the traditional bone-setting group, VAS scores decreased significantly, whereas Constant–Murley scores improved markedly at both follow-up points. The overall outcomes in the traditional bone-setting group were significantly better than those in the surgical group (P<0.05).
Conclusion
Traditional Mongolian bone-setting therapy offers rapid pain relief, superior long-term functional recovery, and fewer complications, making it a valuable treatment option, particularly for elderly patients and those unsuitable for surgery.
4.Research on traditional bone-setting treatment for surgical neck fractures of the humerus
Bolortulga Z ; Jin Ai Hua ; ; Ba Hu Shan ; Ta Na ; Wu Da Mu ; Bao Tu Ya ; Oyuntsetseg N ; Zandi N ; Si Qin ; Tsend-Ayush D
Mongolian Journal of Health Sciences 2026;93(3):178-184
Background:
Mongolian traditional bone-setting is a non-surgical treatment method for fractures; however, there is a lack of evidencebased research evaluating its effectiveness, which forms the basis of our study.
Aim:
To study the therapeutic efficacy of traditional bonesetting treatment for surgical neck fractures of the humerus.
Materials and Methods:
From June 2023 to June 2025, 120 patients diagnosed with surgical neck fractures of the humerus and treated at the Inner Mongolia International Mongolian Hospital were randomly divided into a traditional bone-setting group (n=64) and a surgical treatment group (n=56). Shoulder and upper limb function were evaluated at 1, 3, and 6 months after treatment using the Constant–Murley score, Oxford Shoulder Score (OSS), and Visual Analog Scale (VAS) for pain.
Result:
A total of 120 patients with surgical neck fractures of the humerus were divided into two groups, with no significant differences in the baseline characteristics (p>0.05).In the traditional bone-setting group, pain (VAS) decreased more rapidly at 1–3 months (p<0.01), while activities of daily living and functional outcomes improved significantly at 3–6 months (p<0.01). At 6 months, shoulder function (Constant–Murley score) was significantly improved (p=0.049).In contrast, the surgical group showed faster recovery in joint range of motion and muscle strength in the early stage; however, by 6 months, the outcomes between the two groups were comparable.
Conclusion
In the treatment of surgical neck fractures of the humerus, no significant difference was observed between traditional bone-setting and surgical treatment in terms of shoulder range of motion and muscle strength recovery. However, the advantages of traditional bone-setting lie in its ability to relieve pain more rapidly, promote faster functional recovery, improve activities of daily living, and achieve better long-term outcomes than modern bone-setting. Therefore, traditional bone setting is a safe and effective treatment method that aligns with modern theories of bone healing and rehabilitation principles and holds significant value for wider clinical application and further research in the management of surgical neck fractures of the humerus.
5.Exploration on the differentiation thoughts of external treatment for hemorrhoids based on ancient literature in Zhong Hua Yi Dian
Tingting MU ; Hui XU ; Nina HU ; Yan MA ; Qiujie CAI
International Journal of Traditional Chinese Medicine 2025;47(10):1461-1466
Objective:To explore the external treatment methods of hemorrhoids based on ancient books and documents; To guide clinical medication.Methods:The relevant articles on external treatment of hemorrhoids using Chinese materia medica included in Zhong Hua Yi Dian (5th edition) were retrieved. Excel 2021 was used to analyze the properties, tastes, meridian tropism, external treatment methods and drug efficacy of Chinese materia medica included in the prescriptions, and the relevant discussion contents were sorted out according to the classification of external treatment methods of hemorrhoids. Results:A total of 255 articles in ancient books were included, involving 165 prescriptions. 146 kinds of Chinese materia medica for external treatment of hemorrhoids were selected. The medicinal properties were mainly cold and warm, the main tastes were pungent, bitter and sweet, and the meridian tropism mainly involved the liver, spleen, stomach, heart, lung and large intestine meridians. Fumigation, application and application are the most widely used external treatment methods. Chinese materia medica efficacy mainly includes heat clearing drugs, surface relieving drugs, resuscitation drugs, astringent drugs, anti venom, anti insect and anti itching drugs, and tonic drugs.Conclusions:Ancient physicians treated hemorrhoids with both cold and heat, and paid equal attention to clearing heat and reducing fire, warming and activating blood circulation. They paid attention to the combination of local medication of lesions and the overall treatment of multiple meridians and multiple viscera, so as to achieve the simultaneous treatment of specimens. The treatment methods were rich, including fumigation and washing method to improve local microcirculation, application method using natural adhesive, coating method focusing on precise administration and ligation method of minimally invasive, forming a unique external treatment system, which can be used for modern clinical reference.
6.Analysis of influencing factors of postoperative delirium after endovascular aortic repair: A case-control study
Yi XIONG ; Chuan GAO ; Chunfang MU ; Xuemei ZHAO ; Yuanfei LI ; Chunyue LI ; Kejian HU
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2025;32(10):1462-1466
Objective To analyze the influencing factors of delirium after endovascular aortic repair, and to provide a basis for clinical nursing and prevention of this condition. Methods Patients who underwent endovascular aortic repair at Fuwai Hospital, Chinese Academy of Medical Sciences from 2018 to 2019 were selected. The Chinese version of the Nursing Delirium Screening Scale (Nu-DESC) was used to assess whether postoperative delirium occurred. Patients with a Nu-DESC score≥ 3 were assigned to the delirium group. Non-delirium patients who had the same surgeon and adjacent surgical order were selected at a 1 : 4 ratio to form the non-delirium group. Univariate analysis was performed on the clinical data of the two groups. Factors with P<0.1 in the univariate analysis and those considered clinically significant were included in a multivariate logistic regression analysis to identify the influencing factors of postoperative delirium. Stratified analysis was conducted based on thoracic endovascular aortic repair (TEVAR) and endovascular abdominal aortic repair (EVAR). Results A total of 213 patients were included, comprising 46 in the delirium group and 167 in the non-delirium group. The overall mean age was (60.3±12.0) years, and 183 (85.9%) were male. Univariate analysis showed that emergency admission, preoperative neutrophil percentage, aortic dissection, surgical duration, intubation time, and ICU stay may be associated with postoperative delirium. Multivariate analysis revealed that longer operative and intubation times were associated with a higher likelihood of delirium. In the stratified analysis, the results for the TEVAR group were consistent with the overall findings, whereas no significant differences were observed in the EVAR group. Conclusion Longer surgical and intubation times are associated with an increased risk of delirium in patients undergoing TEVAR. No significant factors influencing delirium are identified in patients undergoing EVAR.
7.Effect of measurement site on diagnostic performance of CT-derived fractional flow reserve
Yutao ZHOU ; Na ZHAO ; Yunqiang AN ; Lei SONG ; Chaowei MU ; Jingang CUI ; Tao JIANG ; Li XU ; Hongjie HU ; Lin LI ; Dumin LI ; Wenqiang CHEN ; Lijuan FAN ; Feng ZHANG ; Yang GAO ; Bin LYU
Chinese Journal of Radiology 2025;59(6):704-711
Objective:To investigate the effect of CT-derived fractional flow reserve (CT-FFR) measurement sites on the values and the diagnostic performance, and to determine the optimal measurement site for CT-FFR using invasive FFR as the reference standard.Methods:This study was part of the CT-FFR CHINA clinical trial. Patients with suspected coronary artery disease who were scheduled for invasive coronary angiography (ICA) were prospectively recruited from five clinical centers across the country from November 2018 to March 2020. Each enrolled patient underwent coronary CT angiography (CCTA), CT-FFR, ICA, and invasive pressure wire-based FFR assessments sequentially within one week. Four groups of CT-FFR values were obtained on each enrolled target vessels according to different CT-FFR measurement locations: 1, 2, 3 cm distal to the target lesion, and terminal vessel groups. Spearman and Bland-Altman analyses were used to explore the correlation and consistency of CT-FFR values and FFR values at different measurement sites. The measurement deviation of CT-FFR was also compared. Diagnostic accuracy and performance of CT-FFR, including sensitivity, specificity, positive predictive value, negative predictive value, and area under the receiver operating characteristic curve (AUC), in discriminating myocardial ischemia were analyzed across all measurement site groups on a per-vessel level, using FFR as the reference standard.Results:A total of 289 patients with 345 target lesion vessels were included. According to CCTA, there were 51 target vessels (14.8%) with<50% stenosis, 106 vessels (30.7%) with 50%-69% stenosis, and 188 vessels (54.5%) with stenosis≥70%. At per-vessel level, CT-FFR and FFR values at each measurement position group were highly positively correlated: 1 cm distal to target lesion group, r=0.734 ( P<0.001); 2 cm distal to target lesion group, r=0.732 ( P<0.001); 3 cm distal to target lesion group, r=0.737 ( P<0.001); terminal vessel group was 0.719 ( P<0.001). At per-vessel level, CT-FFR and FFR values of all measurement sites were in good agreement (Bland-Altman analysis results): 1 cm distal to target lesion group, 0.014 (95% LoA 0.002-0.026); 2 cm distal to target lesion group, 0.026 (95% LoA 0.015-0.038); 3 cm distal to target lesion group, 0.040 (95% LoA 0.039-0.051); terminal vessel group, 0.075 (95% LoA 0.064-0.087). And at per-vessel level, the accuracy of diagnosing myocardial ischemia with CT-FFR at 1 cm was highest [84.6% (95% CI 80.4%-88.3%)], and the lowest accuracy in the terminal vessel group [67.0% (95% CI 61.7%-72.0%)]. However, there was no significant difference in the diagnostic accuracy of CT-FFR at 1 cm, 2 cm [80.6% (95% CI 76.1%-84.6%)] and 3 cm [77.5% (95% CI 72.6%-81.7%)]. AUC of CT-FFR at 1 cm distal to the lesion were both highest for global level and moderately stenosis (50%-69%) lesions [0.85 (95% CI 0.81-0.89), 0.84 (95% CI 0.77-0.90)]. And the differences were statistically significant among the four measurement location groups (all P<0.05). Conclusions:The deviation of CT-FFR increases with measurement site distance distal to target lesions. One centimeter distal to the target lesion is the optimal measurement site, and the CT-FFR value here shows the highest diagnostic performance for myocardial ischemic lesions, especially for moderate stenosis.
8.The application of micro-debridement positioning suture in poor perineal wound healing after delivery
Yongfang MU ; Dan HU ; Maojing ZHAO ; Yangqin CHEN ; Yanyan ZHANG
China Modern Doctor 2025;63(18):14-17
Objective To investigate the clinical effect of micro-debridement and positioning suture in the treatment of poor perineal wound healing after delivery.Methods 106 pregnant women who were eligible for poor healing of perineal wounds(deep skin and muscle defragmentation,involving muscle layer depth ≥1 cm)after vaginal delivery in Affiliated Hospital of Zunyi Medical University were selected as the research objects from June 2021 to October 2024.From June 2021 to December 2022,53 cases in control group were treated with traditional anticipation therapy 1:5000 potassium permanganate solution shallow sitz bath,and from January 2023 to October 2024,53 cases in experimental group were treated with micro-debridement positioning suture.After the operation,follow-up was established with the pregnant women to evaluate the healing cycle of perineal wound,the degree of wound pain 7 days after the operation,the wound healing situation and the cure rate 1 month after the operation.Results The healing period of perineal wound in experimental group was shorter than that in control group;The degree of wound pain in experimental group was lower than that in control group 7 days after operation;The wound healing of one month after operation in experimental group was higher than that in control group;The effective rate of experimental group was higher than that of control group,the differences were statistically significant(P<0.05).Conclusion The use of micro-debridement and positioning suture in the treatment of poor perineal wound can shorten the healing period of perineal wound,reduce the degree of wound pain,the wound healing is good,and the treatment efficiency is good.
9.Study on the mechanistic role of the Fuzheng Huayu formula against cholestatic liver fibrosis in mice
Zheng ZHANG ; Yue LIANG ; Enqi TANG ; Xiaoxi ZHOU ; Yonghong HU ; Gaofeng CHEN ; Wei LIU ; Yongping MU ; Ping LIU ; Jiamei CHEN
Chinese Journal of Hepatology 2025;33(9):889-897
Objective:To investigate the interventional effects of the Fuzheng Huayu (FZHY) formula and its partial mechanistic role on cholestatic liver fibrosis in mice.Methods:Mdr2 gene knockout (Mdr2-/ -) mice were randomly divided into a model group, FZHY group, and Obeticholic acid group. Wild-type C57BL/6J mice of the same age served as the control group. Mdr2-/ -mice were given the corresponding drugs starting from the first day of 9 weeks of age by oral gavage in each group. The control and model groups were administered 0.3% sodium carboxymethylcellulose by oral gavage and were sacrificed at 12 weeks of age for specimen collection. High-speed biochemistry analyzer was used to detect serum alkaline phosphatase and alanine aminotransferase activity in mice. Hematoxylin-eosin staining and Sirius red staining were used to observe pathological changes in liver tissues. Hydroxyproline content was measured to assess collagen in liver tissues. Immunohistochemical staining, Western blotting, and real-time fluorescence quantitative PCR were used to detect the expression of fibrosis markers Col-I and alpha-smooth muscle actin in liver tissues. The expressional condition of cholangiocyte response markers Epcam, CK7, CK19, as well as Pcna, Mki67, and Ccnd1, inflammatory related factors Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4, phosphorylated peroxisome proliferator-activated receptor alpha (PPARα) and nuclear factor kappa-B (NF-κB) were determined. Comparative analysis among multiple groups was performed using one-way ANOVA. The LSD method was used for comparisons between groups. Two-tailed statistical tests were used.Results:Compared with wild-type mice, Mdr2 -/ - mice had a significant increase in serum alanine aminotransferase and alkaline phosphatase activity ( P<0.001). The percentage of Sirius red-positive staining areas and hydroxyproline content in liver tissues was significantly increased ( P<0.01). The expression of Col-I, α-smooth muscle actin, Epcam, CK7, CK19, Pcna, Mki67, and Ccnd1, and the expression of Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4 were significantly increased ( P<0.01); however, both FZHY and Obeticholic acid significantly reversed the increases in these indicators ( P<0.05; P<0.01). Further results showed that compared to wild-type mice, the expression of PPARα was significantly reduced in liver tissues of Mdr2 -/ - mice, while NF-κB was significantly enhanced ( P<0.01). In contrast, compared to Mdr2-/- mice, the expression of PPARα in the liver tissues of FZHY group mice was significantly increased ( P<0.05), while NF-κB was significantly inhibited ( P<0.05). Conclusion:FZHY can significantly improve liver fibrosis, cholangiocyte response, and inflammation in Mdr2 -/ - mice with spontaneously occurring cholestatic liver fibrosis, and its mechanistic role is related to the regulation of the PPARα/NF-κB pathway.
10.Utidelone induces apoptosis and autophagy in small cell lung cancer cells through the ROS/AMPK signaling pathway
Xiaoqian MU ; Chaonan YU ; Yanqiu ZHAO ; Xiufeng HU ; Hongbo WU
Chinese Journal of Oncology 2025;47(8):703-714
Objective:To investigate the effects and underlying molecular mechanisms of Utidelone (UTD1) in small cell lung cancer (SCLC).Methods:The study utilized small cell lung cancer H446 and H1048 cell lines along with animal models. Cell proliferation, cell cycle progression, apoptosis, autophagy, and related activities following UTD1 treatment were assessed using Cell Counting Kit-8 (CCK-8), flow cytometry, immunofluorescence staining, reactive oxygen species (ROS) generation assay, and Western blot analysis. The involvement of the ROS/adenosine monophosphate-activated protein kinase (AMPK) signaling pathway was also examined. Data analysis was performed using GraphPad Prism version 8 software.Results:UTD1 inhibited the viability of H446 and H1048 cells in a dose- and time-dependent manner. The half inhibitory concentrations (IC 50) of UTD1 for H446 and H1048 cells were 0.675 and 0.439 μg/ml, respectively. The proportion of cells in the G 2/M phase for H446 and H1048 cells in the UTD1 group at 6 h, 12 h, and 24 h was [(53.86±4.54)%, (68.59±5.49)%, (60.89±3.26)%] and [(46.83±2.20)%, (60.67±3.44)%, (57.88±5.11)%], which were significantly higher than that in the control group, except for the proportion of H1048 cells at 6 h [(38.99±2.60)% vs. (40.73±2.50)%, P<0.05]. The apoptosis rates were [(23.57±0.12)%, (35.79±1.59)%, and (46.15±4.57)%] for H446 cells and [(23.05±2.70)%, (37.73±2.97)%, and (43.39±3.31)% for H1048 cells], all of which were significantly higher than those in the control group [(6.44±0.96)%, (6.31±0.75)%, respectively; all P<0.05]. The number of LC3 fluorescent spots was [(56±11), (69±8), and (66±8)] for H446 cells and [(39±7), (56±12), and (50±11)] for H1048 cells, both significantly higher than those in the control group [(13±6) and (12±5), respectively; both P<0.05]. The relative fluorescence intensity of ROS was 2.54±0.48, 2.85±0.68, and 5.03±0.72 for H446 cells and 2.26±0.51, 4.17±0.35, and 4.66±0.51 for H1048 cells, which were also significantly higher than those in the control group ( P<0.05). The expression levels of cyclin B1, cyclin A2, and P21 of H446 cells in the three time points were [(0.63±0.07, 0.33±0.05, 0.23±0.04), (0.68±0.08, 0.46±0.03, 0.27±0.06), and (0.64±0.03, 0.32±0.05, 0.22±0.03), respectively], all significantly lower compared to the control group ( P<0.05). The apoptosis rates of H446 and H1048 cells in the UTD1+Z-VAD-FMK group were (19.97±3.19)% and (17.68±3.14)%, both lower than those in the UTD1 group [(40.73±3.35)% and (39.82±2.45)%, respectively; all P<0.05]. The absorbance values of H446 and H1048 cells in the UTD1+3-MA group were significantly higher than those in the UTD1 group at 6h, 12h, and 24h (all P<0.05). The levels of p-AMPKα/AMPKα, LC3-II expression, and the percentage of apoptotic cells in the H446 and H1048 cells of the UTD1+NAC group were [(1.33±0.09, 1.33±0.11), (1.49±0.16, 1.55±0.05), (17.24±2.15)%, and (19.40±4.28)%], all of which were lower than those observed in the UTD1 group [(1.98±0.17, 2.23±0.23), (2.81±0.19, 2.49±0.38), (38.07±3.53)%, and (41.20±1.87)%, all P<0.05]. The number of LC3 fluorescence points and the percentage of apoptotic cells in the H446 and H1048 cells of the UTD1+si-AMPKα group [(24±5, 23±3), (18.35±1.15)%, and (19.15±3.46)%] were all lower than those in the UTD1+si-NC group [(46±6, 36±6), (39.34±1.77)%, and (39.50±2.15)%, all P<0.05]. The tumor inhibition rates in small cell lung cancer tumor-bearing nude mice for the 2.5 mg/kg UTD1 group and the 5 mg/kg UTD1 group were 46.43% and 58.33%, respectively. Furthermore, the proportions of apoptosis-positive cells and p-AMPKα-positive cells in the UTD1 group were significantly higher compared to the control group, while the levels of Ki-67 positivity were significantly reduced. Conclusion:UTD1 inhibits SCLC cell proliferation, induces G 2/M phase arrest, and promotes cell apoptosis and autophagy through the activation of the ROS/AMPK signaling pathway.

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