1.Effect of miR-196b-5p in adipose-derived stem cell exosomes on burn wound healing in rats
Na ZUO ; Qi TANG ; Meng YU ; Kai TAO
Chinese Journal of Tissue Engineering Research 2026;30(1):43-49
BACKGROUND:Currently,miR-196b-5p has been found to play a role in cell proliferation,migration and inhibition of scar hyperplasia,but there is a lack of relevant studies on whether it plays a role in the process of wound healing.OBJECTIVE:To investigate the effect of miR-196b-5p in adipose stem cell-derived exosomes on burn wound healing.METHODS:The models of deep second degree skin burn in SD rats were established and randomly divided into four groups:blank control group,exosome group,agomiR-196b-5p group,and exosome+antagomiR-196b-5p group,with 10 rats in each group.PBS,adipose-derived stem cell-derived exosomes,miR-196b-5p agonist,and miR-196b-5p inhibitor were injected around the wound according to different groups.Wound healing was observed immediately after injury and on days 7,14,and 21 after injury.On day 7 after surgery,hematoxylin-eosin staining was used to observe the expression of inflammation in the wound surface.On day 14 after suregery,Masson staining was used to observe the expression of collagen and immunohistochemical staining was used to observe the expression of CD31 in the wound surface.On day 7 after surgery,western blot assay was performed to detect the expression of α-smooth muscle actin and type Ⅰ collagen in the wound surface.RESULTS AND CONCLUSION:(1)The wound healing was faster in the agomiR-196b-5p group,while it was slower in the blank control group and the exosome+antagomiR-196b-5p group.(2)Compared with the blank control group and the exosome+antagomiR-196b-5p group,the wound infiltration of inflammatory cells was less in the exosome group and the agomiR-196b-5p group,and the expression of CD31 was significantly increased(P<0.01).(3)Compared with the blank control group and the exosome+antagomiR-196b-5p group,the expression levels of α-smooth muscle actin and type Ⅰ collagen were increased in the exosome group and the agomiR-196b-5p group(P<0.05).These findings indicate that miR-196b-5p in adipose stem cell-derived exosomes can promote burn wound healing in rats.
2.Thread embedding pretreatment at Xinshu(BL 15)improves cardiac function of acute myocardial ischemia rats
Xiaoqing CHEN ; Luyao BIAN ; Xingyu LU ; Tao YANG ; Li Xiang HAI
Chinese Journal of Tissue Engineering Research 2026;30(4):882-891
BACKGROUND:Acupuncture at Xinshu(BL 15)can significantly improve cardiac function and protect myocardial cells in acute myocardial ischemia,but the effect and mechanism of thread embedding treatment at Xinshu(BL 15)on cardiac function in acute myocardial ischemia are yet unclear.Nuclear factor κB activation often appears as an intranuclear translocation of the P65 isoform,and activation of the nuclear factor κB signaling pathway is marked by elevated P65 levels.OBJECTIVE:To explore the effects of thread embedding pretreatment at Xinshu(BL 15)on cardiac function and the expression levels of interleukin-10,tumor necrosis factor-α,P65 genes and proteins in rats with acute myocardial ischemia.METHODS:Thirty-two male Sprague-Dawley rats were randomly divided into a blank group,a model group,a Xinshu(BL 15)acupoint group,and a non-meridian/non-acupoint group using a random number table method,with eight rats in each group.Rat models of acute myocardial ischemia were established in the latter three groups.The Xinshu(BL 15)acupoint group had thread embedding at Xinshu(BL 15)for 14 days,followed by subcutaneous injection of isoproterenol hydrochloride into the back to establish an acute myocardial ischemia rat model.The non-meridian/non-acupoint group had local thread embedding for 14 days,and the rest procedures were the same as above.In the model group,Xinshu(BL 15)was only marked,and the rest procedures were the same as above.In the blank group,Xinshu(BL 15)was only marked,and then an equal amount of physiological saline was injected subcutaneously into the back.After 24 hours of modeling,electrocardiogram and cardiac ultrasound were performed.Abdominal aorta blood was extracted for detection of serum creatine kinase and creatine kinase isoenzyme levels using enzyme-linked immunosorbent assay.Subsequently,the rats were euthanized and samples were collected.Hematoxylin-eosin and TUNEL staining were used to observe the pathological changes of myocardial tissue and the apoptosis of myocardial cells.Real-time fluorescence quantitative PCR(RT-qPCR)and western blot were used to detect the mRNA and protein expression of tumor necrosis factor-α,interleukin-10,and P65 in myocardial tissue respectively.RESULTS AND CONCLUSION:(1)Electrocardiogram:Compared with the blank group,the model group,non-meridian/non-acupoint group,and Xinshu(BL 15)acupoint group had significantly elevated ST segment in lead Ⅱ of the electrocardiogram.(2)Cardiac ultrasound:Compared with the model group,the Left ventricular end-systolic dimension in the Xinshu(BL 15)acupoint group were significantly reduced(P<0.05),while left ventricular ejection fraction and left ventricular fractional shortening rate were significantly increased(P<0.05).(4)Serum creatine kinase and creatine kinase isoenzyme:Compared with the model group,the Xinshu(BL 15)acupoint group showed a significant decrease in serum creatine kinase and creatine kinase isoenzyme levels(P<0.05).(4)Hematoxylin-eosin staining:Compared with the model group,the arrangement of myocardial fibers in the Xinshu(BL 15)acupoint group was basically neat,with less edema and a small amount of inflammatory cell infiltration.(5)TUNEL staining:Compared with the model group,the fluorescence intensity of myocardial cell apoptosis in the Xinshu(BL 15)acupoint group was significantly reduced,and its apoptosis rate was significantly reduced(P<0.05).(6)RT-qPCR and western blot:Compared with the model group,the myocardial tissue interleukin-10 level in the Xinshu(BL 15)acupoint group was significantly increased(P<0.05),while tumor necrosis factor-α and P65 levels were significantly decreased(P<0.05).These findings indicate that thread embedding pretreatment at Xinshu(BL 15)can improve cardiac function in rats with acute myocardial ischemia,and its mechanism of action may be related to the inhibition of the activation of the nuclear factor-κB signaling pathway.
3.Mitogen-activated protein kinase signaling pathway regulates the development of osteoarthritis:guiding targeted therapy with traditional Chinese medicine
Hao LI ; Hongcheng TAO ; Ping ZENG ; Jinfu LIU ; Qiang DING ; Chicheng NIU ; Kai HUANG ; Hongyu KANG
Chinese Journal of Tissue Engineering Research 2026;30(6):1476-1485
BACKGROUND:Osteoarthritis is pathologically characterized by progressive degeneration of the articular cartilage and abnormal deformation of the subchondral bone.In recent years,with the deepening of medical research,it has been found that the mitogen-activated protein kinases(MAPK)signaling pathway has a regulatory role in inflammatory cell infiltration,inflammatory factor release,and chondrocyte proliferation,which is particularly important for the treatment of osteoarthritis.OBJECTIVE:To briefly review the main research progress in the mechanism of MAPK signaling pathway regulating osteoarthritis in recent years,aiming to provide new ideas for the treatment of osteoarthritis.METHODS:CNKI,WanFang and PubMed databases were searched for relevant literature using the search terms of"mitogen-activated protein kinases,osteoarthritis,extracellular signal-regulated MAP kinases,p38 mitogen-activated protein kinases,JNK mitogen-activated protein kinase"in Chinese and English.Relevant literature published from January 2019 to November 2024 was searched,and 108 articles were finally included for summary analysis.RESULTS AND CONCLUSION:(1)Various stimuli inside and outside the cells activate the MAPK signaling pathway,regulate gene transcription and protein synthesis,and promote the release of inflammatory factors,such as tumor necrosis factor-α,interleukin-1β,and interleukin-6.The release of these inflammatory factors aggravates the progression of osteoarthritis.(2)The active ingredients of traditional Chinese medicine,mainly saponins and flavonoids,as well as Chinese herbal formulas and preparations with the main effects of activating blood circulation and removing blood stasis,tonifying the liver and kidney,can play a therapeutic role in osteoarthritis by inhibiting the MAPK signaling pathway,regulating the release of matrix metalloproteinases,balancing the homeostatic state of osteogenesis and osteoblastogenesis,attenuating the synovial inflammation,decreasing the release of inflammatory factors and inflammatory vesicles,decreasing cellular pyroptosis,promoting autophagy,and ameliorating oxidative stress.(3)Although traditional Chinese medicine has become popular in the treatment of osteoarthritis by virtue of its own advantages of multi-components,multi-targets,multi-pathways,and low side effects,the use of MAPK signaling pathway to guide the treatment of individual osteoarthritis is the difficulty of the technology,which needs to be continuously researched and explored.(4)Therefore,further development of relevant herbal inhibitors that can modulate the MAPK signaling pathway may be a potential drug strategy for the treatment of osteoarthritis in the future.
4.Establishment and validation of a model for femoral head necrosis after internal fixation of femoral neck fracture using logistic regression and SHAP analysis
Long LIAO ; Zepeng ZHAO ; Zongyuan LI ; Qinglong YU ; Tao ZHANG ; Jinyuan TANG ; Nan YE ; Han XU ; Bo SHI
Chinese Journal of Tissue Engineering Research 2026;30(3):626-633
BACKGROUND:The most common complication of traumatic femoral neck fractures after internal fixation is femoral head necrosis.Currently,many studies have reported on the risk factors that affect the occurrence and development of postoperative femoral head necrosis,but there is still a lack of tools to predict the risk of femoral head necrosis after internal fixation of femoral neck fractures.OBJECTIVE:To develop a predictive model that estimates the risk of femoral head necrosis shortly after patients with femoral neck fractures receive cannulated screw internal fixation.METHODS:A retrospective analysis reviewed clinical records of 172 patients who underwent cannulated screw internal fixation for femoral neck fractures at Department of Orthopedics of Mianyang Central Hospital from January 2013 to June 2023.Patients were categorized into two groups based on the presence or absence of femoral head necrosis within one year post-operation:the necrosis group and the non-necrosis group.Univariate analysis,Lasso regression,and multivariate Logistic regression techniques were employed to identify the determinants of femoral head necrosis.A nomogram prediction model was constructed using R language's"rms"package,version 4.0.The receiver operating characteristic curve was used to evaluate the discriminatory ability of the model.The Hosmer-Lemeshow test was used to evaluate the goodness of fit of the model,and the decision curve analysis was used to determine its clinical application benefits.Internal validation of the study was conducted using the Bootstrap method,involving 1 000 repeated samplings.To delve deeper into the primary factors influencing femoral head necrosis post-internal fixation of the femoral neck,this paper employed the SHAP method for data set analysis.RESULTS AND CONCLUSION:(1)The risk factors leading to femoral head necrosis in the short term after cannulated screw fixation of femoral neck fractures include:smoking,diabetes,Garden classification,fracture line location,reduction quality,age,and operation time.(2)The prediction model demonstrated robust performance,evidenced by an area under the curve of 0.940(95%Confidence Interval:0.903 to 0.977),indicating a high level of prediction accuracy.The model achieved a sensitivity of 90.2%and a specificity of 87.6%,indicating that its diagnostic performance was stable.The Hosmer-Lemeshow goodness-of-fit test yielded a chi-square value of 6.593 with a P-value of 0.581,confirming that the model's predictions closely align with the observed outcomes.(3)The calibration curve of the model also performed well,and its overall trend was very close to the ideal curve,further proving the high accuracy of the model.(4)The internal validation was carried out by the Bootstrap method with 1 000 repeated samplings,and the area under the curve of the model internal validation was still as high as 0.939,proving that the model had good stability.(5)Through the decision curve,it is found that within the probability threshold range of 1%to 92%,the model can obtain the maximum net benefit value.(6)The SHAP analysis results show that among the risk factors analyzed in this study,the location of the fracture line serves as the most significant predictor of femoral head necrosis following internal fixation with cannulated screws in femoral neck fractures,and subcapital fractures are extremely prone to femoral head necrosis after surgery.(7)It is concluded that the validated prediction model demonstrates strong discriminative power and reliability,offering practical clinical utility.It serves as a useful reference tool for short-term risk assessment of femoral head necrosis following internal fixation of femoral neck fractures.
5.Effect of sacroiliac joint ankylosis on outcomes of L5/S1 transforminal lumbar interbody fusion and lumbar sagittal parameters
Yalei WANG ; Xuezhi WANG ; Tao ZHOU ; Xinxin SHEN ; Ding FANG ; Hongliang CHEN
Chinese Journal of Tissue Engineering Research 2026;30(3):634-641
BACKGROUND:The correlation between sacroiliac joint degeneration and lumbar degenerative disease has been analyzed in the literature in the past,but the clinical efficacy and imaging changes after interbody fusion with sacroiliac joint ankylosis in patients with lumbar degenerative disease have not been reported in the literature.OBJECTIVE:To investigate the effect of sacroiliac joint ankylosis on the clinical efficacy and lumbar sagittal regression after L5/S1 single-segment transforminal lumbar interbody fusion in patients with lumbar degenerative disease.METHODS:Thirty-seven patients who underwent L5/S1 segmental transforminal lumbar interbody fusion for lumbar degenerative disease with sacroiliac joint ankylosis between June 2020 and September 2023 in Affiliated Hospital of Xuzhou Medical University were retrospectively analyzed as group A.Thirty-seven patients with lumbar degenerative disease without sacroiliac joint ankylosis who were matched for general information during the same period were selected as controls in group B.Clinical efficacy was assessed using the Oswestry disability index and visual analog scale for lumbar and lower limb pain.The lumbar sagittal parameters included lumbar anterior convexity angle,lumbar partial anterior convexity angle,and lower lumbar anterior convexity angle.Pfirrmann grading was used to assess the degree of preoperative disc degeneration,postoperative endplate damage and screw loosening,and to record the fusion of the operated segments at the final postoperative follow-up visit.RESULTS AND CONCLUSION:(1)There was no statistically significant difference in age,body mass index,bone mineral density,operation time,intraoperative bleeding,preoperative primary diagnosis and postoperative follow-up time between the two groups(P>0.05).(2)The preoperative Pfirrmann grading of lumbar disc degeneration in group A patients(3.4±0.9)was significantly higher than that of group B(3.1±0.6),and the difference was statistically significant(t=2.059,P=0.044).(3)All patients showed significant improvement in postoperative lumbar sagittal parameters compared with preoperative ones(all P<0.05).During the follow-up period,there was a loss of correction in patients in group A.There was no statistical difference in the lumbar anterior convexity angle,lower lumbar anterior convexity angle,and local anterior convexity angle at the last follow-up compared with the preoperative period(P>0.05).The lumbar anterior convexity angle,lower lumbar anterior convexity angle,and local anterior convexity angle in group A were significantly lower than those of group B patients at both preoperative and final follow-up,and the differences were statistically significant(all P<0.05).(4)There was no statistically significant difference in postoperative endplate injury between the two groups(x2=0.181,P=0.670),and screw loosening was significantly higher in group A than in group B,with a statistically significant difference(x2=4.163,P=0.041).(5)At the last follow-up,the incidence of grade 3 fusion and grade 4 fusion was significantly higher in group A than in group B.The difference in the distribution of fusion grades between the two groups was statistically significant(x2=7.848,P=0.031).(6)The Oswestry disability index and lower limb visual analog scale scores at the last follow-up of both groups were significantly improved compared with the preoperative period(P<0.05).The visual analog scale scores for low back pain at 3 months after surgery and at the last follow-up of group A were significantly higher than those of group B(t=2.010,P=0.048;t=2.133,P=0.036).(7)It is concluded that regardless of whether it is accompanied by sacroiliac joint ankylosis or not,lumbar degenerative disease patients who undergo interbody fusion with foramen magnum can achieve good therapeutic effects,but lumbar degenerative disease patients with sacroiliac joint ankylosis who undergo interbody fusion with foramen magnum at the L5/S1 segments have a poorer improvement of low back pain than patients without sacroiliac joint ankylosis after the operation.Furthermore,patients with preoperative sacroiliac ankylosis who underwent L5/S1 segmental transforminal lumbar interbody fusion had a low fusion rate and were prone to loss of correction of the lumbar sagittal position.
6.Effects of resistance training on quadriceps mass and knee joint function in patients with osteoporosis and sarcopenia
Jian ZHOU ; Tao ZHANG ; Weili ZHOU ; Xingcheng ZHAO ; Jun WANG ; Jie SHEN ; Li QIAN ; Ming LU
Chinese Journal of Tissue Engineering Research 2026;30(5):1081-1088
BACKGROUND:The quadriceps strength of patients with osteoporosis and sarcopenia is significantly reduced,which can further reduce the function of the knee joint,affect the function of the lower limbs and even lead to a decrease in whole-body coordination.It is speculated that a reasonable quadriceps training program and personalized guidance are beneficial to the recovery of knee joint function in patients with osteoporosis and sarcopenia.OBJECTIVE:To observe the effect of short-term moderate-intensity resistance rehabilitation training on the mass and function of the quadriceps and knee joint function in patients with osteoporosis and sarcopenia.METHODS:Using the integrated physical examination and rehabilitation model,375 patients with osteoporosis and sarcopenia were screened at the Health Management Center of Shanghai Public Health Clinical Center.They underwent 12 weeks of combined/comprehensive exercise rehabilitation based on resistance exercise,including quadriceps resistance isotonic and isometric contraction training twice a week(3-5 sets each time,10-15 minutes per set)and aerobic exercise/balance exercise two or three times a week(30 minutes each time).Assessments and data collection were performed before rehabilitation training,12 weeks after rehabilitation training,and at follow-up 12 weeks after stopping rehabilitation training,mainly including knee joint range of motion and proprioception,quadriceps muscle strength,and cross-sectional area(magnetic resonance imaging results),pain,knee joint function(Hospital for Special Surgery score)and walking function("up-and-go"time and 6 m pace test results)as well as the patient's psychological status assessment.RESULTS AND CONCLUSION:All 375 patients completed 12 weeks of rehabilitation training and 12 weeks of follow-up without any adverse events.(1)Compared with before training,the patients' gait speed and knee range of motion increased significantly after 12 weeks of rehabilitation training(P<0.01),the time of"stand-to-walk"decreased(P<0.01),and the proprioception of the knee joint and the strength of the quadriceps femoris were significantly improved(P<0.01);and at the follow-up visit 12 weeks after stopping training,the above indicators and functions of the patients were well maintained(P>0.05).(2)Magnetic resonance imaging results showed that the effective cross-sectional area of the quadriceps femoris did not improve significantly after 12 weeks of rehabilitation training(P>0.05);but the Hospital for Special Surgery score of knee joint function increased significantly(P<0.01),and the visual analog pain scale score decreased significantly(P<0.01),suggesting that this may be related to the improvement of quadriceps femoris quality by resistance rehabilitation training.(3)The results of the Hospital Anxiety and Depression Scale score showed that the anxiety and depression scores of the patients continued to decrease,both at 12 weeks of rehabilitation training and at 12 weeks after stopping training(P<0.01).It is suggested that resistance rehabilitation training of the quadriceps can help patients with osteoporosis and sarcopenia to restore quadriceps muscle strength,increase range of motion,improve proprioception and joint stability,thereby enhancing knee joint function,reducing pain,improving depression and anxiety,and to a certain extent promoting the coordinated recovery of the musculoskeletal system.
7.Acute effects of blood flow restriction in low-intensity resistance training on endothelial function-related inflammatory factors
Chinese Journal of Tissue Engineering Research 2026;30(5):1184-1195
BACKGROUND:Long-term blood flow restriction combined with low-intensity resistance training has been shown to effectively treat obesity by alleviating chronic inflammation and endothelial dysfunction.However,the immediate effects of a single session on serum concentrations of vascular endothelial function and inflammatory biomarkers remain unclear.OBJECTIVE:To explore the short-term effects and recovery capacity of blood flow restriction during low-intensity resistance training on serum biomarkers of vascular endothelial function and inflammation in obese male college students.METHODS:Twenty obese male college students(body mass index>30 kg/m2,body fat percentage>25%)were randomly assigned to a control group(0%arterial occlusion pressure)or a blood flow restriction group(80%arterial occlusion pressure).Both groups performed a single session of low-intensity resistance training at an intensity corresponding to a perceived exertion of 11-13 on the Rate of Perceived Exertion Scale.The training was repeated three times,with each session lasting 30 minutes,totaling 1.5 hours.Serum biomarkers were measured before exercise,immediately post-exercise,1 hour post-exercise,and 24 hours post-exercise.The assessed biomarkers included vascular endothelial function markers,inflammatory markers,and insulin function indicators.RESULTS AND CONCLUSION:(1)Vascular endothelial function:Acute exercise increased vascular endothelial growth factor A concentrations in both groups.The blood flow restriction group significantly elevated serum platelet-derived growth factor and nitric oxide levels(P<0.05),while the control group showed a significant increase in nitric oxide synthase levels(P<0.05).Angiotensin Ⅱ concentrations decreased immediately after acute exercise in both groups but remained significantly higher than baseline in the blood flow restriction group after 24 hours of recovery,and there was a significant difference between the two groups(P<0.05).(2)Regarding inflammatory markers,the blood flow restriction group induced higher levels of hypoxia and significantly upregulated tumor necrosis factor-α and hypoxia-inducible factor-1α concentrations(P<0.05).Adiponectin and leptin levels upregulated in both groups,with a more pronounced rise in adiponectin level in the blood flow restriction group than the control group(P<0.05).lnterleukin-6 concentrations decreased in both groups,with a greater reduction in the blood flow restriction group.(3)For insulin function,the blood flow restriction and control groups showed immediate increases and decreases in insulin levels after exercise,respectively,but these returned to below and above baseline levels after 24 hours of recovery.Both groups reduced insulin resistance index in adipose tissue,with a more significant improvement in the blood flow restriction group(P<0.05).To conclude,compared with low-intensity resistance training,short-term blood flow restriction induces more favorable changes in inflammatory and vascular endothelial biomarkers,improving inflammation and endothelial dysfunction with longer-lasting effects.However,further studies are needed to validate these findings over long-term interventions.
8.Exploring Therapeutic Mechanism of Xihuang Wan for Hyperplasia of Mammary Glands Based on Network Pharmacology, Molecular Docking, and Cell Experiments
Junliang WANG ; Peihua LIANG ; Xueli MA ; Juanxia SUN ; Tao HAN ; Yongmei LAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):41-49
ObjectiveTo study the therapeutic mechanism of Xihuang Wan for hyperplasia of mammary glands based on network pharmacology, molecular docking, and cell experiments. MethodsThe active ingredients and targets of Xihuang Wan were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and A Bioinformatics Annotation daTabase for Molecular mechANism of Traditional Chinese Medicine (BATMAN-TCM) and supplemented by searching against PubChem and Swiss Target Prediction. The targets of differential metabolites in tissues and urine were obtained from previous metabolomics studies through PubChem and Swiss Target Prediction. GeneCards, Online Mendelian Inheritance in Man (OMIM), PharmGKB, Therapeutic Target Database (TTD), Drunbank were searched for the targets of hyperplasia of mammary glands. After the common targets were obtained via Veeny2.1.0, the STRING database was used to analyze the protein-protein interactions, and Cytoscape was used for the core target analysis and visualization. Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were employed for enrichment analysis. Molecular docking was carried out in Autodock, and cell experiments were conducted to verify the prediction results. In the cell experiments, estradiol and progesterone (E2+P) were used to intervene in human mammary epithelial/MCF-10A cells, and thus the MCF-10A cell proliferation model was established. The cells were then treated with Xihuang Wan-medicated serum. The cell counting kit-8 (CCK-8) was used to measure the cell proliferation, and flow cytometry was used to detect apoptosis. The mRNA and protein levels of key factors in MCF-10A cells were determined by real-time PCR and Western blot, respectively. ResultsThe results of network pharmacology showed that 90 active ingredients and 316 common targets were obtained, from which 20 core targets and 38 corresponding active ingredients were screened out. The results of GO and KEGG enrichment analyses showed that Xihuang Wan exerted effect against hyperplasia of mammary glands by regulating a variety of biological processes, which may be related to protein kinase B (Akt)-related molecular functions, estrogen signaling pathway, prolactin signaling pathway and other biological processes. The results of molecular docking showed that estrogen receptor 1 (ESR1), serine/threonine kinase 1 (Akt1), non-receptor tyrosine kinase (SRC), and signal transducer and activator of transcription 3 (STAT3) all had strong binding activity with the nine active ingredients, suggesting that Xihuang Wan exert the effect through the ESR1/SRC/phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway and the Janus kinase (JAK)/STAT3 signaling pathway. The results of cell experiments showed that E2+P intervention in MCF-10A cells promoted the proliferation of MCF-10A cells (P<0.05), while the Xihuang Wan-medicated serum inhibited the proliferation of MCF-10A cells exposed to E2+P (P<0.05). Flow cytometry showed that the Xihuang Wan-medicated serum promoted the apoptosis of MCF-10A cells exposed to E2+P (P<0.01). The results of Real-time PCR showed that the Xihuang Wan-medicated serum down-regulated the mRNA levels of PI3K, Akt, JAK2, and STAT3 in MCF-10A cells treated with E2+P (P<0.01). The results of Western blot showed that the Xihuang Wan-medicated serum inhibited the expression of p-PI3K/PI3K, p-Akt/Akt, p-JAK2/JAK2, and p-STAT3/STAT3 in MCF-10A cells treated with E2+P (P<0.05). ConclusionXihuang Wan may exert the effect against hyperplasia of mammary glands by inhibiting the proliferation and promoting the apoptosis of MCF-10A cells, which may related to the inhibition of the activation of PI3K/Akt and JAK2/STAT3 signaling pathways.
9.Analysis of Differential Metabolites of Pinelliae Rhizoma at Different Browning Stages Based on Widely Targeted Metabolomics
Jing TAO ; Honghong LIANG ; Ruoshi LI ; Zhouli XU ; Minzhao LI ; Aien TAO ; Guihua JIANG ; Li AI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):191-199
ObjectiveTo investigate differential metabolites associated with browning in the post-harvest processing of Pinelliae Rhizoma, providing data support for elucidating the key metabolites and metabolic pathways involved in browning, and developing safe and efficient sulfur-free processing techniques. MethodsUltra-performance liquid chromatography-triple quadrupole/linear ion trap mass spectrometry(UPLC-QTRAP-MS/MS) was used to detect the metabolites of Pinelliae Rhizoma at different browning stages(0, 8, 16 h) for widely targeted metabolomics. Subsequently, Multivariate statistical analysis of metabolites was conducted using principal component analysis(PCA), hierarchical cluster analysis(HCA), orthogonal partial least squares-discriminant analysis(OPLS-DA), and K-means cluster analysis. Differential metabolites at different browning stages were screened based on variable importance in the projection(VIP) value>1 and |log2fold change(FC)|≥1, and metabolic pathway enrichment analysis was performed using Kyoto Encyclopedia of Genes and Genomes(KEGG). ResultsA total of 1 416 metabolites were identified across the three browning stages of Pinelliae Rhizoma, predominantly comprising amino acids and their derivatives(239), lipids(219), alkaloids(156), phenolic acids(121), terpenoids(113), and flavonoids(111). A two-by-two comparison of the three browning phases, yielded 622 differential metabolites that were significantly enriched in the phenylpropanoid biosynthesis, flavone and flavonol biosynthesis, and purine metabolic pathway. Further analysis revealed that carbohydrates such as D-mannose and turanose, phenolic acids such as 1-O-caffeoyl-6-O-glucosyl-β-D-glucose, dicaffeoylshikimic acid, and flavonoids such as epigallocatechin gallate, vitexin-7-O-rutinoside, luteolin-7-O-(6″-malonyl)glucoside-5-O-arabinoside, catechin gallate, epicatechin gallate, isovitexin-7-O-glucoside-2″-O-rhamnoside, apigenin-7-O-rutinoside-4ʹ-O-sophoroside, 3,5,3ʹ,4ʹ,5ʹ-penta-hydroxyflavan-7-gallate may act as browning substrates and play important roles in the browning process. ConclusionCarbohydrates, phenolic acids, and flavonoids may serve as key substrates in the browning process of Pinelliae Rhizoma, involving pathways such as phenylpropanoid biosynthesis, flavone and flavonol biosynthesis, and purine metabolism, which can provide a theoretical basis for further exploration of the browning mechanism.
10.Molecular Mechanisms of Salvia Miltiorrhiza and Its Active Ingredients against Colorectal Cancer: A Review
Jianing GUO ; Xiaochen NI ; Kaiyuan ZHANG ; Wei FAN ; Chuhang WANG ; Chao XU ; Jianbo HUANG ; Tao JIANG ; Guangji ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):307-314
Colorectal cancer (CRC) is one of the most common cancers, with its incidence ranking high among cancers. It stands as the second leading cause of cancer-related death worldwide. In the early stages, CRC lacks specific symptoms, and most patients are diagnosed at advanced stages, making it a major research focus in the field of gastrointestinal tumors. Currently, clinical CRC treatments face several common challenges, including high surgical risks, frequent metastasis and recurrence, drug resistance, and significant side effects from chemotherapy and radiation therapy. With the development and application of traditional Chinese medicine (TCM), it has been found that TCM and its active ingredients can effectively inhibit CRC cell proliferation, invasion, migration, and angiogenesis, and promote apoptosis and autophagy, thereby slowing the progression of CRC. This has become a key focus of CRC treatment research. Salvia Miltiorrhiza has multiple pharmacological effects, including activating blood circulation to dispel blood stasis, unlocking meridians to relieve pain, clearing heat to calm irritability, and cooling blood to reduce abscesses. It contains a variety of chemical components, including diterpenoids, phenolic acids, flavonoids, polysaccharides, nitrogen-containing compounds, steroids, and lactone compounds. This review summarized the molecular mechanisms of Salvia miltiorrhiza and its active ingredients in the treatment of CRC. It is found that these ingredients exert anti-CRC effects through various molecular mechanisms, including cell cycle arrest, promotion of apoptosis, inhibition of cell invasion and migration, induction of autophagy, suppression of tumor angiogenesis, and remodeling of the tumor microenvironment. The review aims to provide new insights for the drug development and clinical application of Salvia miltiorrhiza in CRC treatment.

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