1.Bioinformatics Reveals Mechanism of Schisandrin B in Inhibiting Ferroptosis to Ameliorate Methionine and Choline Deficiency-induced Fatty Liver Disease in Mice
Zhifeng ZHU ; Wenting LI ; Yongjun CAO ; Yuanyuan LIN ; Yifei LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):74-83
ObjectiveNonalcoholic fatty liver disease (NAFLD) is a metabolic stress liver injury. Ferroptosis is involved in the occurrence and development of NAFLD. Exploring the efficacy and mechanism of schisandrin B in treating NAFLD facilitates the development of strategies for the prevention and treatment of NAFLD. MethodsThe molecular structure of schisandrin B was obtained by searching against PubChem, and the related targets were predicted by SwissTargetPrediction. The active ingredients and their targets were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and the high-throughput experiment- and reference-guide database of traditional Chinese medicine (HERB). GeneCards and FerrDb were searched for the targets of NAFLD and ferroptosis. The common targets were taken as the core targets, and the protein-protein interaction network of the core targets was established. DAVID was used for gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Finally, molecular docking was performed between schisandrin B and core targets, and the binding energy was calculated. C57BL/6 mice were fed with a methionine and choline-deficiency (MCD) diet for the modeling of NAFLD. Mice were randomized into normal, model, positive drug (essentiale), and low- and high-dose schisandrin B groups. The body mass and liver index of mice were measured after drug administration. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum and those of total cholesterol (TC), triglyceride (TG), malondialdehyde (MDA), glutathione (GSH), and Fe2+ in the liver homogenate were measured by biochemical assay kits. The pathological changes of the liver tissue were observed by hematoxylin-eosin (HE) and red oil O staining. Enzyme-linked immunosorbent assay was employed to determine the levels of interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF)-α, and 4-hydroxynonenal (4-HNE) in the serum. Western blotting and real-time PCR were employed to determine the protein and mRNA levels, respectively, of solute carrier family 7 member 11 (SLC7A11), solute carrier family 3 member 2 (SLC3A2), glutathione peroxidase 4 (GPX4), transferrin, and ferritin heavy chain (FTH) in the liver tissue. ResultsA total of 2 370, 2 547, and 1 451 targets of schisandrin B, NAFLD, and ferroptosis were obtained, in which 90 common targets were shared by the three. Enrichment analyses predicted 505 GO terms and 92 KEGG pathways. Molecular docking suggested that schizandrin B had strong binding affinity with the key targets of ferropstosis (SLC7A11 and SLC3A2). Animal experiments showed that schizandrin B significantly decreased the liver index, lowered the levels of ALT, AST, TC, TG, IL-6, IL-1β, and TNF-α, alleviated hepatocyte ballooning and inflammatory cell infiltration, and reduced lipid accumulation in the liver of NAFLD mice. In addition, schisandrin B significantly lowered the levels of MDA, 4-HNE, and Fe2+, elevated the level of GSH, up-regulated the protein and mRNA levels of SLC7A11, SLC3A2, and GPX4, and down-regulated the protein and mRNA levels of transferrin in the liver tissue. ConclusionSchisandrin B can alleviate NAFLD by inhibiting ferroptosis in hepatocytes.
2.Exploring Role of Energy Dyshomeostasis in Metabolic Dysfunction-associated Fatty Liver Disease Panvasculopathy from Theory of Liver Being Substantial Yin and Functional Yang
Jing CUI ; Qian XU ; Wenting WANG ; Mengmeng ZHU ; Yanfei LIU ; Yue LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):227-233
Liver being substantial Yin and functional Yang maintain normal function of Qi, blood and meridians. In clinical practice, it is often found that pan-vascular lesions with atherosclerosis as the predominant pathological change often co-occur with metabolic dysfunction-associated fatty liver disease(MAFLD). MAFLD leads to increased risk and worse prognosis for many pan-vascular diseases, including cardiovascular disease. Dysregulation of energy homeostasis disrupts the hepatic homeostasis of body use, and representative drugs to improve metabolism, such as metformin, sodium-glucose co-transporter 2 inhibitors, and glucagon-like peptide-1 agonists, not only have a clear cardiovascular benefit, potential improvement of MAFLD has also been demonstrated. The liver stores blood and the heart pumps blood, and liver diseases affect the heart, that's why the unsmoothness of vessels appears. So the treatment should from the standpoint of liver, restoring liver function, soothing the liver and nourishing heart, activating blood and dredging meridian. It is of great significance to explore in depth the pathogenesis and treatment of pan-vascular lesions caused by MAFLD, and to restore the energy homeostasis by adjusting the balance of liver Yin and Yang.
3.Exploring Role of Energy Dyshomeostasis in Metabolic Dysfunction-associated Fatty Liver Disease Panvasculopathy from Theory of Liver Being Substantial Yin and Functional Yang
Jing CUI ; Qian XU ; Wenting WANG ; Mengmeng ZHU ; Yanfei LIU ; Yue LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):227-233
Liver being substantial Yin and functional Yang maintain normal function of Qi, blood and meridians. In clinical practice, it is often found that pan-vascular lesions with atherosclerosis as the predominant pathological change often co-occur with metabolic dysfunction-associated fatty liver disease(MAFLD). MAFLD leads to increased risk and worse prognosis for many pan-vascular diseases, including cardiovascular disease. Dysregulation of energy homeostasis disrupts the hepatic homeostasis of body use, and representative drugs to improve metabolism, such as metformin, sodium-glucose co-transporter 2 inhibitors, and glucagon-like peptide-1 agonists, not only have a clear cardiovascular benefit, potential improvement of MAFLD has also been demonstrated. The liver stores blood and the heart pumps blood, and liver diseases affect the heart, that's why the unsmoothness of vessels appears. So the treatment should from the standpoint of liver, restoring liver function, soothing the liver and nourishing heart, activating blood and dredging meridian. It is of great significance to explore in depth the pathogenesis and treatment of pan-vascular lesions caused by MAFLD, and to restore the energy homeostasis by adjusting the balance of liver Yin and Yang.
4.Application value of prediction model based on magnetic resonance imaging machine learning algorithm and radiomics in predicting lymphovascular invasion status of rectal cancer with-out lymph node metastasis
Leping PENG ; Xiuling ZHANG ; Yuanhui ZHU ; Ling WANG ; Wenting MA ; Yaqiong MA ; Gang HUANG ; Lili WANG
Chinese Journal of Digestive Surgery 2024;23(8):1099-1111
Objective:To construct an prediction model based on magnetic resonance imaging (MRI) machine learning algorithm and radiomics and investigate its application value in predicting lymphovascular invasion (LVI) status of rectal cancer without lymph node metastasis.Methods:The retrospective cohort study was conducted. The clinicopathological data of 204 rectal cancer patients without lymph node metastasis who were admitted to Gansu Provincial Hospital from February 2016 to January 2024 were collected. There were 123 males and 81 females, aged (61±7)years. All 204 patients were randomly divided into the training dataset of 163 cases and the testing dataset of 41 cases by a ratio of 8∶2 using the electronic computer randomization method. The training dataset was used to construct the prediction model, and the testing dataset was used to validate the prediction model. The clinical prediction model, radiomics model and joint prediction model were constructed based on the selected clinical and/or imaging features. Measurement data with normal distribution were represented as Mean± SD. Count data were described as absolute numbers, and the chi-square test or Fisher exact probability were used for comparison between the groups. Comparison of ordinal data was conducted using the nonparameter rank sum test. The inter-class correlation coefficient (ICC) was used to evaluate the consistency of the radiomics features of the two doctors, and ICC >0.80 was good consistency. Univariate analysis was conducted by corres-ponding statistic methods. Multivariate analysis was conducted by Logistic stepwise regression model. The receiver operating characteristic (ROC) curve was drawn, and the area under the curve (AUC), Delong test, decision curve and clinical impact curve were used to evaluate the diagnostic efficiency and clinical utility of the model. Result:(1) Analysis of factors affecting LVI status of patients. Of the 204 rectal cancer patients without lymph node metastasis, there were 71 cases with positive of LVI and 133 cases with negative of LVI. Results of multivariate analysis showed that gender, platelet (PLT) count and carcinoembryonic antigen (CEA) were independent factors affecting LVI status of rectal cancer without lymph node metastasis in training dataset [ odds ratio=2.405, 25.062, 2.528, 95% confidence interval ( CI) as 1.093-5.291, 2.748-228.604, 1.181-5.410, P<0.05]. (2) Construction of clinical prediction model. The clinical prediction model was conducted based on the results of multivariate analysis including gender, PLT count and CEA. Results of ROC curve showed that the AUC, accuracy, sensitivity and specificity of clinical prediction model were 0.721 (95% CI as 0.637-0.805), 0.675, 0.632 and 0.698 for the training dataset, and 0.795 (95% CI as 0.644-0.946), 0.805, 1.000 and 0.429 for the testing dataset. Results of Delong test showed that there was no significant difference in the AUC of clinical prediction model between the training dataset and the testing dataset ( Z=-0.836, P>0.05). (3) Construction of radiomics model. A total of 851 radiomics features were extracted from 204 patients, and seven machine learning algorithms, including logistic regression, support vector machine, Gaussian process, logistic regression-lasso algorithm, linear discriminant analysis, naive Bayes and automatic encoder, were used to construct the prediction model. Eight radiomics features were finally selected from the optimal Gaussian process learning algorithm to construct a radiomics prediction model. Results of ROC curve showed that the AUC, accuracy, sensitivity and specificity of radiomics prediction model were 0.857 (95% CI as 0.800-0.914), 0.748, 0.947 and 0.642 for the training dataset, and 0.725 (95% CI as 0.571-0.878), 0.634, 1.000 and 0.444 for the testing dataset. Results of Delong test showed that there was no significant difference in the AUC of radiomics prediction model between the training dataset and the testing dataset ( Z=1.578, P>0.05). (4) Construction of joint prediction model. The joint prediction model was constructed based on the results of multivariate analysis and the radiomics features. Results of ROC curve showed that the AUC, accuracy, sensitivity and specificity of radiomics prediction model were 0.885 (95% CI as 0.832-0.938), 0.791, 0.912 and 0.726 for the training dataset, and 0.857 (95% CI as 0.731-0.984), 0.854, 0.714 and 0.926 for the testing dataset. Results of Delong test showed that there was no significant difference in the AUC of joint prediction model between the training dataset and the testing dataset ( Z=0.395, P>0.05). (5) Performance comparison of three prediction models. Results of the Hosmer-Lemeshow goodness-of-fit test showed that all of the clinical prediction model, radiomics prodiction model and joint prediction model having good fitting degree ( χ2=1.464, 12.763, 10.828, P>0.05). Results of Delong test showed that there was no signifi-cant difference in the AUC between the clinical prediction model and the joint prediction model or the radiomics model ( Z=1.146, 0.658, P>0.05), and there was a significant difference in the AUC between the joint prediction model and the radiomics model ( Z=2.001, P<0.05). Results of calibra-tion curve showed a good performance in the joint prediction model. Results of decision curve and clinical impact curve showed that the performance of joint prediction model in predicting LVI status of rectal cancer without lymph node metastasis was superior to the clinical prediction model and the radiomics model. Conclusions:The clinical prediction model is constructed based on gender, PLT count and CEA. The radiomics predictive model is constructed based on 8 selected radiomics features. The joint prediction model is constructed based on the clinical prediction model and the radiomics predictive model. All of the three models can predict the LVI status of rectal cancer with-out lymph node metastasis, and the joint prediction model has a superior predictive performance.
5.GC Fingerprint and Multi-component Content Determination of Xiangsha Yangwei
Yiqing MAO ; Liqing TONG ; Ying ZHU ; Qing ZHANG ; Wenting LIU ; Yu CAO
Herald of Medicine 2024;43(1):115-121
Objective To establish the method of fingerprint and content determination of multi-component for Xiangsha Yangwei pill by gas chromatography(GC).Methods The GC fingerprint of Xiangsha Yangwei pill was found,and the peak attribution was carried out.The contents of limonene,eucalyptol,camphor,borneol,bornyl acetate,patchouli alcohol,pogostone,and α-cyperone were determined.Results The fingerprint similarity of 56 batches of Xiangsha Yangwei pill were 0.33-0.99,28 common peaks were confirmed,and 14 known components were identified.Limonene,eucalyptol,camphor,borneol,bornyl acetate,patchouli alcohol,pogostone and α-cyperone showed good linearity within the determined ranges(14.30-286.08,24.52-490.44,16.14-322.88,9.40-187.95,15.39-307.83,25.78-515.60,19.95-398.90,and 24.87-497.30 μg·mL-1).The average recoveries were 101.20%,97.90%,93.97%,94.23%,102.94%,100.54%,99.16%,and 98.31%;with the RSDs were 2.41%,1.48%,1.65%,2.00%,1.93%,2.30%,2.07%,and 2.38%,respectively.The concentrations of eight components were 0.2-959.1,0.3-420.4,1.0-542.6,0.0-64.5,0.0-364.2,0.0-339.6,0.0-130.7,0.0-82.0 μg·g-1,respectively.Conclusion The fingerprint and multi-component determination method can be used for the quality control and evaluation of Xiangsha Yangwei pill.
6.Relationship Between Gut Microbiota Dysbiosis and Cardiometabolic Diseases Based on "Heart-spleen-intestine" Axis
Qian XU ; Wenting WANG ; Yiwen LI ; Jing CUI ; Mengmeng ZHU ; Yanfei LIU ; Yue LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(20):203-211
Cardiometabolic disease is a clinical syndrome with a causal relationship between metabolic abnormalities and cardiovascular damage. With its global incidence and related mortality rates continually rising, it has become a major health concern worldwide. The role of the gut microbiome and its metabolic products in cardiovascular metabolic health has received widespread attention, with gut microbiota dysbiosis considered a key factor in promoting the development of cardiometabolic disease. Dysbiosis disrupts the balance of the "heart-spleen-intestine" axis, leading to dysfunction of the spleen and intestines, which triggers metabolic disorders and accelerates the progression of cardiometabolic disease. Cardiac dysfunction can also negatively affect spleen and intestinal function, leading to imbalances in the heart and spleen, disharmony in Qi and blood, and exacerbating metabolic anomalies and further dysbiosis, thus forming a vicious cycle. From a modern biological perspective, the gut microbiome and its metabolic products can influence disease progression by modulating inflammatory responses and immune imbalances, leading to endothelial dysfunction and metabolic disorders, thereby increasing the risk of cardiometabolic disease. Additionally, the article proposes strategies for managing cardiometabolic disease by regulating the gut microbiome through a combination of Chinese and western medicine approaches. Traditional Chinese medicine(TCM) treatment starts from the gut microbiome, using the "heart-spleen-intestine" axis as a mediator to regulate cardiovascular metabolic health, highlighting the unique advantages of TCM in targeting the gut microbiome to treat cardiometabolic disease. This article takes the TCM theory of the "heart-spleen-intestine" axis as a starting point, discusses the pivotal role played by this axis in the connection between gut microbiome dysbiosis and the development of cardiometabolic disease, aiming to provide a new perspective for the integrated traditional Chinese and western medical research on cardiometabolic disease, offering scientific evidence and practical guidance to improve the prognosis and quality of life for patients.
7.Correlations of brain network functional connectivity alterations with cerebrospinal fluid pathological markers in patients with Alzheimer's disease
Chengbing GONG ; Zhengyang ZHU ; Jingxian XU ; Wenting SONG ; Haifeng CHEN ; Ruomeng QIN ; Hui ZHAO ; Yun XU
Chinese Journal of Neuromedicine 2024;23(1):18-26
Objective:To explore the correlations of brain network functional connectivity (FC) alterations with cerebrospinal fluid (CSF) pathological biomarkers in patients with Alzheimer's disease (AD).Methods:A total of 39 patients with cognitive impairment, admitted to Department of Neurology, Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University from January 2020 to December 2022 were recruited; 23 patients were with AD and 16 with non-AD. Clinical data were compared between the 2 groups. Resting-state functional MRI (rs-fMRI) data were collected, and FC differences between brain networks and FC differences within brain networks were compared by independent component analysis. Correlations of FC differences between brain networks and FC differences within brain networks with concentrations of β-amyloid protein 1-42 (Aβ 1-42) and Tau protein in CSF were analyzed. Results:Compared with the non-AD group, AD group had significantly lower Aβ 1-42 in CSF ( P<0.05). Compared with those in the non-AD group, FC alterations between the left frontoparietal network (lFPN) and anterior default mode network (aDMN) and between the visual network (VN) and posterior cingulate cortex (PCC), as well as FC alterations in lFPN, were significantly increased in AD group ( P<0.05). Compared with those in the non-AD group, FC alterations between lFPN and cerebellar network (CEN), and FC alterations in aDMN, sensorimotor network (SMN) and VN were significantly decreased in AD group ( P<0.05). In AD group, FC in SMN was positively correlated with total Tau and phosphorylated-Tau181 in CSF ( P<0.05); FC between VN and PCC was positively correlated with total Tau in CSF ( P<0.05). CSF Aβ 1-42 was positively correlated with FC alterations in aDMN and VN, but negatively correlated with FC in FPN ( P<0.05). Conclusion:In AD patients, characteristic changes in FC within and between multiple brain networks are noted, which are related to changes of Tau protein and Aβ 1-42 in CSF.
8.Effects of breastfeeding on immune response of CD4+T lymphocytes in infants
Simin ZHU ; Wenjuan TU ; Wenting ZHANG ; Ziqi TU ; Cheng′an WANG
Chinese Journal of Child Health Care 2024;32(1):103-107
【Objective】 To explore the effects of breastfeeding on the immune response of CD4+T lymphocytes in infants in non-inflammatory state, and to analyze the immunomodulatory significance of the whole composition of breast milk. 【Methods】 A retrospective cohort study was conducted. From January to September 2022, six-month-old infants who took physical examination in the Child Healthcare Department of Changzhou Children′s Hospital Affiliated to Nantong University, were selected based on inclusion criteria, and were divided into breastfeeding group (n=33) and formula feeding group (n=27) based on their feeding patterns. Flow cytometry was used to detect the percentage of CD4+ T cells, including helper T cell (Th) 1, Th2, Th17, regulatory T cell (Treg), and the levels of related cytokines interleukin (IL)-2, IL-4, IL-6, IL-10, tumor necrosis factor (TNF)-α, interferon (IFN)-γ, IL-17 in peripheral blood. The differences in these indicators between the two groups were compared. 【Results】 Compared with the formula feeding group, the breastfeeding group showed significantly higher percentages of Th1(t=3.038), Treg (t=2.088). The ratio of Th1 to Th2(Z=2.756), IL-10(Z=2.297) and IFN-γ (Z=2.076) in the peripheral blood of the breastfeeding group were also significantly higher. Conversely, the breastfeeding group had significantly lower percentage of Th17(Z=2.704) and IL-17A (t=2.187) (P<0.05). There was no significant difference the percentage of Th2, as well as in the levels of IL-2, IL-4, IL-6 and TNF-α between the two groups (P>0.05). 【Conclusions】 Breastfeeding has a regulatory effect on the immune response of infant CD4+ T lymphocytes. It promotes the development of Th1/Th2 towards Th1 and the immunomodulatory effect of Treg. Moreover, it inhibits the Th17 type immune response. These findings suggest that the complete composition of breast milk contributes to the development and maturation of infant immune system, enhancing immune defense and immune tolerance.
9.Regulation of Immune Balance by Traditional Chinese Medicine in Treatment of Cough Variant Asthma: A Review
Han YANG ; Yonghuang YAN ; Wenting ZHANG ; Peixuan ZHU ; Fang YAN ; Yujie WU ; Shiqing QIAO ; Tieshan WANG ; Zeqi SU ; Ting WANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):206-212
Cough variant asthma (CVA) is a chronic respiratory disease with cough as its main symptom. The occurrence of CVA is closely related to non-specific airway inflammation, and its pathogenesis involves environmental, genetic, immune, and other factors. In recent years, the advantages of traditional Chinese medicine (TCM) in the treatment of CVA have attracted the attention of experts and scholars in China and abroad, especially its prominent role in regulating immune balance, relieving cough symptoms in CVA patients, and reducing recurrence. T Helper cells 1 (Th1), T helper cells 2 (Th2), T helper cells 17 (Th17), and regulatory T cells (Treg) are derived from CD4+ T cells. Immune imbalance of Th1/Th2 and Th17/Treg is a new hotspot in the pathogenesis of CVA and a potential key target in the treatment of CVA by TCM. Th cell subsets are in dynamic balance under physiological conditions, maintaining respiratory immune homeostasis in which pro-inflammatory cytokines and anti-inflammatory cytokines are balanced. Immature helper T cells (Th0) can be differentiated into Th1, Th2, Th17, Treg, and other cell subsets due to cytokine types in the microenvironment in the stage of CVA maturation. The proliferation of Th2 cells leads to eosinophilic airway inflammation. Excessive differentiation of Th17 cells induces neutrophil airway inflammation. Th1/Th2 and Th17/Treg cells are mutually restricted in number and function, and the immune imbalance of Th1/Th2 and Th17/Treg is easy to aggravate the generation of inflammatory response. Restoring immune balance is particularly important for the airway anti-inflammatory therapy of CVA. In this paper, the imbalance of Th1/Th2 and Th17/Treg and the pathogenesis of CVA were systematically expounded. Meanwhile, the latest research on the regulation of immune imbalance by TCM compound, single TCM, and its effective ingredients in the treatment of CVA was reviewed. It provides ideas and references for revealing the scientific connotation of TCM regulating immune balance therapy of CVA, as well as the development of clinical treatment and basic research of CVA.
10.Effect of volatile oil of Ligusticum chuanxiong on transdermal properties and cytotoxicity of triptolide in vitro
Cheng JIANG ; Zhenzhong ZANG ; Lihua CHEN ; Haiting ZHU ; Shihua FU ; Weifeng ZHU ; Wenting WU ; Wei SHI ; Yongmei GUAN
China Pharmacy 2024;35(4):413-418
OBJECTIVE To explore the effect of volatile oil of Ligusticum chuanxiong on the transdermal properties and cytotoxicity of triptolide in vitro. METHODS The chemical constituents of the volatile oil of L. chuanxiong were analyzed by gas chromatography-mass spectrometry. The lower abdominal skin of KM mice was separated and divided into triptolide group, triptolide in compatibility with volatile oil of L. chuanxiong groups at 1∶10, 1∶50, 1∶100 (hereinafter referred to as “compatibility 1∶10”“compatibility 1∶50”“compatibility 1∶100” groups). After the skin of mice in each group was fully exposed to 0.2 g of the corresponding cream for 24 h, the cumulative transdermal dose (Qn) of triptolide in the receiving solution of each group was determined by high-performance liquid chromatography, and the transdermal absorption rate (Jss) was calculated. Human immortalized keratinocytes (HaCat) were used as a model, the CCK-8 method was used to detect the cell survival rate of different concentrations of the volatile oil of L. chuanxiong and triptolide before and after compatibility. RESULTS A total of 62 chemical constituents of the volatile oil of L. chuanxiong were identified, including Z-ligustilide, senkyunolide, and β-selinene. The Qn (P< 0.01) and Jss of triptolide increased within 24 h in the compatibility 1∶10 and 1∶50 groups, while the Qn (P<0.05) and Jss decreased in the compatibility 1∶100 group as compared with the triptolide group. Compared with the triptolide group, the cell survival rate of HaCat was significantly increased in the compatibility 1∶10 and 1∶50 groups when the triptolide concentrations were 36, 72 and 144 ng/mL (P<0.05 or P<0.01); while the cell survival rate of HaCat was decreased in the compatibility 1∶100 group, but the difference was not statistically significant (P>0.05). CONCLUSIONS When the compatibility ratio of triptolide and volatile oil of L. chuanxiong was 1∶10 or 1∶50, it can promote the transdermal absorption of triptolide and reduce the cytotoxicity of triptolide to HaCat.

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